EPA-600/3-78-021
February 1978
Ecological Research Series
THE BIOENVIRONMENTAL IMPACT
OF A COAL-FIRED POWER PLANT
Third Interim Report
Colstrip, Montana
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Corvallis, Oregon 97330
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This report has been assigned to the ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal spe-
cies, and materials. Problems are assessed for their long- and short-term influ-
ences. Investigations include formation, transport, and pathway studies to deter-
mine the fate of pollutants and their effects. This work provides the technical basis
for setting standards to minimize undesirable changes in living organisms in the
aquatic, terrestrial, and atmospheric environments.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/3-78-021
February 1978
THE BIOENVIRONMENTAL IMPACT
OF A COAL-FIRED POWER PLANT
Third Interim Report, Colstrip, Montana
December 1977
Edited by
Eric M. Preston and Robert A. Lewis
Ecological Effects Research Division
Corvallis Environmental Research Laboratory
Corvallis, OR 97330
CORVALLIS ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CORVALLIS, OR 97330
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DISCLAIMER
This report has been reviewed by the Corvallis Environmental Research Labora-
tory, U.S. Environmental Protection Agency, and approved for publication.
Mention of trade names or commercial products does not constitute endorsement
or recommendation for use.
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FOREWORD
Effective regulatory and enforcement actions by the Environmental
Protection Agency would be virtually impossible without sound scientific data
on pollutants and their impact on environmental stability and human health.
Responsibility for building this data base has been assigned to EPA's Office of
Research and Development and its 15 major field installations, one of which is
the Corvallis Environmental Research Laboratory (CERL).
The primary mission of the Corvallis Laboratory is research on the effects
of environmental pollutants on terrestrial, freshwater, and marine ecosystems;
the behavior, effects and control of pollutants in lake systems; and the
development of predictive models on the movement of pollutants in the bio-
sphere.
The Colstrip, Coal-fired Power Plant Project is the first attempt to
generate methods to predict the bioenvironmental effects of air pollution
before damage is sustained. The methods are to be framed within an overall
facility siting strategy for coal-fired power plants in the northern plains.
This will permit planners to assess the ecological impact of energy conversion
activities on grasslands prior to site selection. If environmental impacts can
be predicted, mitigation efforts can be designed to minimize the adverse
effects of power plant construction and operation.
A. F. Bartsch
Director, CERL
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ABSTRACT
The Environmental Protection Agency (EPA) has recognized the need for a
rational approach to the incorporation of ecological impact information into
power facility siting decisions in the northern great plains. The Colstrip
Coal-fired Power Plant Project (CFPPP) is one aspect of the agency's response
to this need. Research funded by the CFPPP is a first attempt to generate
methods to predict the bioenvironmental effects of air pollution before
damage is sustained. The methods are to be framed within an overall facility
siting strategy for coal-fired power plants in the northern plains. This
document describes the progress of the investigation.
Pre-construction documentation of the environmental characteristics of
the grassland ecosystem in the vicinity of Colstrip, Montana began in the
summer of 1974. This documentation continued until Colstrip generating unit
1 began operation in September, 1975. Since then, key characteristics of
the ecosystem have been monitored regularly to detect possible pollution
impacts upon plant and animal community structure; primary production, inver-
tebrate animal consumers, and decomposers; plant and animal diseases; both
beneficial and harmful insects; indicators and predictors of pollution (e.g.,
lichens and honeybees); physiological responses of plants and vertebrate
animals; insect behavior (mainly of honeybees) and production; the behavior,
reproduction and development, population biology, health and condition of
vertebrate animals.
In the summer of 1975, field stressing experiments were begun to provide
the data necessary to develop dose-response models for S02 stress on a grass-
land ecosystem. These experiments involve continuous stressing of one acre
grassland plots with measured doses of S02 during the growing season (usually
April through October). Results of the 1975 field season investigations are
summarized in this publication. The six-year project will terminate in 1980
and a final report will be published after data analyses are complete.-
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CONTENTS
Page
Foreword iii
Abstract iv
List of Contributors vii
Acknowledgements ix
INTRODUCTION
Assessment of Ecological Impact from the Operation of a
Coal-Fired Power Plant in the Northcentral Great Plains
R. A. Lewis, E. M. Preston, N. R. Glass 2
Sections COLSTRIP STUDIES
1 Effects of S02 and other Coal-Firing Plant Emissions on Pro-
ducer, Invertebrate Consumer, and Decomposer Structure and
Function in the Vicinity of Col strip, Montana
W. K. Lauenroth, J. L. Dodd, R. K. Heitschmidt,
J. W. Leetham 13
2 Plant Community Studies in the Vicinity of Colstrip, Montana
J. E. Taylor, W. C. Leininger 41
3 Soil and Epiphytic Lichen Communities of the Colstrip,
Montana Area
S. Eversman 50
4 Investigation of the Impact of Coal-Fired Power Plant Emis-
sions upon the Disease/Health/Growth Characteristics of
Ponderosa Pine-Skunkbush Ecosystems and Grassland Ecosystems
in Southeastern Montana
C. C. Gordon, P. C. Tourangeau, P. M. Rice 65
5 Investigation of the Impact of Coal-Fired Power Plant Emis-
sions upon Insects: Entomological Studies in the Vicinity
of Colstrip, Montana
J. J. Bromenshenk 140
6 The Effects of Coal-Fired Power Plant Emissions on Vertebrate
Animals in Southeastern Montana (A report of progress)
R. A. Lewis, M. L . Morton, M. D. Kern, J. D. Chilgren,
E. M. Preston 213
7 Remote Sensing of the Bioenvironmental Effects of Stack
Emissions in the Colstrip Vicinity
J. E. Taylor, W. C. Leininger 280
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Sections Page
8 Integrated Aerosol Characterization Monitoring, Colstrip,
Montana
N. L. Abshire, V. E. Derr, G. T. McNice, R. Pueschel,
C. Van Valin 291
FIELD AND LABORATORY EXPERIMENTS
9 Zonal Air Pollution System: Design and Performance
J. J. Lee, R. A. Lewis 322
10 First-year Effects of Controlled Sulfur Dioxide Fumigation on
a Mixed-Grass Prairie Ecosystem
J. L . Dodd, W. K. Lauenroth, R. K. Heitschmidt,
J. W. Leetham 345
11 Monitoring Plant Community Changes Due to S02 Exposure
J. E. Taylor, W. C. Leininger 376
12 Effects of Low-Level S02 Stress on Two Lichen Species
S. Eversman 385
13 Effects of Low-Level S02 Exposure on Sulfur Accumulation and
Various Plant Life Responses of Some Major Grassland Species
Under Field Conditions
C. C. Gordon, P. M. Rice, P. C. Tourangeau 399
14 Investigation of the Impact of Coal-Fired Power Plant Emis-
sions upon Insects: Entomological Studies at the Zonal Air
Pollution System
J. J. Bromenshenk 473
15 The Relative Sensitivity of Selected Plant Species to Several
Pollutants Singly and in Combination
D. T. Tingey, L. Bard, R. W. Field 508
16 Progress in Modeling the Effects of S02 Fumigation on an
Eastern Montana Grassland
J. L. Dodd, M. Coughenour, W. K. Lauenroth 514
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LIST OF CONTRIBUTORS
N. L. Abshire
U.S. Dept. of Commerce
N.O.A.A.
Environmental Research Laboratories
Boulder, Colorado 80302
L. Bard
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
J. J. Bromenshenk
Environmental Studies Laboratory
University of Montana
Missoula, Montana 59801
J. D. Chilgren
U.S. EPA/CERL
200 S.W. 35th St.
Con/all is, Oregon 97330
M. Coughenour
Natural Resources Ecology Laboratory
Colorado State University
Fort Collins, Colorado 80521
V. E. Derr
U.S. Dept. of Commerce
N.O.A.A.
Environmental Research Laboratories
Boulder, Colorado 80302
J. L. Dodd
Natural Resources Ecology Laboratory
Colorado State University
Fort Collins, Colorado 80521
S. Eversman
Dept. of Biology
Montana State University
Bozeman, Montana 59715
R. W. Field
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
N. R. Glass
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
C. C. Gordon
Environmental Studies Laboratory
University of Montana
Missoula, Montana 59801
R. K. Heitschmidt
Natural Resources Ecology Laboratory
Colorado State University
Fort Collins, Colorado 80521
M. D. Kern
Dept. of Biology
The College of Wooster
Wooster, Ohio 44691
W. K. Lauenroth
Natural Resources Ecology Laboratory
Colorado State University
Fort Collins, Colorado 80521
J. J. Lee
U.S. EPA/CERL
200 S.W. 25th St.
Corvallis, Oregon 97330
J. W. Leetham
Natural Resources Ecology Laboratory
Colorado State University
Fort Collins, Colorado 80521
W. C. Leininger
Dept. of Animal and Range Sciences
Montana State University
Bozeman, Montana 59715
R. A. Lewis
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
VII
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G. T. McNice
U.S. Dept. of Commerce
N.O.A.A.
Environmental Research Laboratories
Boulder, Colorado 80302
M. L. Morton
Dept. of Biology
Occidental State College
1600 Campus Rd.
Los Angeles, California 90041
E. M. Preston
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
R. Pueschel
U.S. Dept of Commerce
N.O.A.A.
Environmental Research Laboratories
Boulder, Colorado 80302
P. M. Rice
Environmental Studies Laboratory
University of Montana
Missoula, Montana 59801
J. E. Taylor
Dept. of Animal and Range Sciences
Montana State University
Bozeman, Montana 59715
D. T. Tingey
U.S. EPA/CERL
200 S.W. 35th St.
Corvallis, Oregon 97330
P. C. Tourangeau
Environmental Studies Laboratory
University of Montana
Missoula, Montana 59801
C. Van Valin
U.S. Dept. of Commerce
N.O.A.A.
Environmental Research Laboratories
Boulder, Colorado 80302
vm
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ACKNOWLEDGEMENTS
Many individuals have contributed to the preparation of this document.
Editoral assistance was generously provided by Ms. Karen Raldolph. The help of
Mr. Thomas Hill and others is much appreciated.
Our work could not proceed without the help and support of the people of
southeastern Montana, especially the ranchers on whose land we are working and
the personnel and persons residing at and near Fort Howes Ranges Station,
Custer National Forest. The Kluver's and the McRae's have been particularly
supportive. '
IX
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INTRODUCTION
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ASSESSMENT OF ECOLOGICAL IMPACT FROM THE
OPERATION OF A COAL-FIRED POWER PLANT IN THE
NORTHCENTRAL GREAT PLAINS
by
R. A. Lewis, E. M. Preston, and N. R. Glass
INTRODUCTION
The principal short-term constraints on the utilization of coal reserves
of the northcentral United States are the amount of environmental degradation
that the American public is willing to sustain as the price for secure and
abundant energy and the ability of scientists to forecast the amount and kinds
of environmental impacts that will result from a given level of coal use.
Because of the rapid expansion of coal mining and coal-fired power
generation in the northcentral great plains and because of the very limited
knowledge and experience with the responses of the range resource to air
pollution and other impacts of coal development, it is important to assess the
potential responses of these ecosystems. These grasslands exhibit limited
resilience to disturbance, and there is thus a potential for irreversible
effects. It is not now possible to predict the long-term consequences of even
the present level of coal production in the west (Lawton and McNeil, 1973;
Library of Congress, 1975).
One of the primary concerns of the Colstrip, Montana, Coal-fired Power
Plant Project (CFPPP) is the development of methods for the predictive
assessment of impact. If this project is successful, the ability to
judiciously plan regional power plant inventories and to select sites for new
power production facilities will be greatly enhanced. Furthermore, if the
nature and extent of specific impact can be predicted, steps to minimize or to
ameliorate impacts may be possible. If the predictions are accepted, it should
be simpler to resolve conflicts among parties concerned with the siting process
and to arrive at constructive agreements that reduce the overall costs of power
plant siting.
PREDICTIVE ASSESSMENT OF IMPACT
Federal legislation requires the prediction of impact whether the
capability exists or not. The National Environmental Policy Act of 1969 (NEPA)
requires that an environmental impact statement (EIS) be filed for any project
that requires federal approval. The EIS is to identify and describe potential
environmental impacts of such projects and to evaluate their significance.
NEPA, however, provides only limited guidelines for the conduct and preparation
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of the EIS. Consequently, most EISs provide few data that are useful to
decision makers or that satisfy the intent of the law.
The problems of evaluating and forecasting environmental consequences and
developing the technological methods to reduce or ameliorate impact are complex,
and_CFPPP contributions at this time are little more than embryonic. Both strip
mining and coal combustion produce effects on living and non-living components
of the environment that are extensive and diverse. While some effects may be
ephemeral (acute toxicity of sulfur dioxide, fluoride, nitrogen oxides, etc.),
others may prove to be insidious and long-lasting or irreversible (acid
rainfall, long-term soil fertility decline, decreased energy flow, retrograde
succession, etc.).
AVAILABLE FORECASTING TOOLS
Forecasting tools that have been more or less routinely employed (e.g.,
guessing, extrapolation or rationalization of retrospective information, etc.)
have had very limited or unknown success, because follow-up on predictions is
infrequent. Those tools that have the greatest potential for predicting
environmental impacts on terrestrial ecosystems (e.g., the use of controlled in
situ field stressing with pollutants of concern combined with appropriate
laboratory experiments; simulation models such as mass balance models; models of
simple food webs; and diffusion models that employ data from the sites or
regions of concern) are rarely employed.
Four basic types of predictive tools can be identified. These are
qualitative techniques, retrospective analyses of data and trend extrapolation,
process models, and empirical methods coupled to predictive models. Qualitative
methods are based upon judgments and intuition that are logically arrived at in
the absence of any satisfactory theory, direct experience or quantitative data
relating to the problem. Such methods may lead to conceptual models that may
indicate what data are needed and can generate hypotheses that can be tested.
Qualitative methods include Delphi methods, panel (workshop) concensus,
intuition and visionary forecast, and historical analysis. Occasionally,
historical analogy (which is applied widely in EISs) proves useful for planning
horizons of several years.
The use of scenarios, while not a predictive technique, should also be
mentioned. Scenarios are hypothetical narratives that explore potential
alternatives, usually those that seem reasonable or possible (worst case, best
case). They are speculative, but can provide some insights into the range of
possible future states of the system and can force consideration of options that
might otherwise be overlooked.
Methods that employ retrospective data and trend analysis may be very
effective where relevant data are fairly complete for one or more decades and
when the functions of interest are noise-free and stable or follow an obvious
trend. The basic assumption underlying these methods is that the observed
patterns will continue into the future. Consequently, such methods tend to be
better predictors of specific short-term futures than of long-term trends.
Since such methods are based only upon time series analysis and extrapolation,
the relationships of environmental variables to trends are unknown. As a
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result, predictions are highly site and time specific. A further serious
drawback to the use of retrospective data is that it typically includes
information that is not standardized, or is incomplete. Such data must be used
selectively and this provides excellent opportunity for the introduction of
social, economic or political bias.
Process models can be powerful predictive tools. While costly to develop,
such models have a good potential for long-term prediction if they are based
upon the results of appropriate field and laboratory experiments that are
designed to evaluate the effects of random and non-random variables on the
output.
The environmental residual technique has also been used in environmental
impact assessment. This technique could be much improved by a more thorough
assessment of impact than is usual. Undesirable effects (residuals) of power
generation can be determined through a detailed knowledge of the power plant
operations. One dimension of a two-dimensional matrix defines each residual.
The other dimension lists management strategies. By filling out the cells or
elements within this matrix, it is possible to optimize the management of the
power plant based upon any one or all of the residuals of interest. Different
management methods might prove to be most effective for different types of
pollutants. For example, it is possible that SO could best be managed by
building a tall stack or by intermittent controls, while management of heavy
metals might be best accomplished through the construction and operation of a
baghouse. The detail of each management strategy relative to each residual
would in turn be dependent upon engineering design and properties of the
particular coal which was to be mined. The main difficulty is in relating the
residuals to a detailed environmental impact assessment which can in turn feed
back upon the initial design and site location of the power plant so as to
minimize impacts.
For the environmental residuals method to be effectively applied, a
detailed impact assessment with predictive capabilities is required. Residual
allocation models have been linked to models that determine resource
availability based on economics as well as geography and end uses of the
particular energy activity from which residuals are generated (Federal Power
Commission, in press). Briefly, this is accomplished through modeling that
depends upon a knowledge of the resource being exploited, the technology being
used, the chemical and other residuals which are produced by the technology,
and the economics at each stage. These factors can be used to generate a matrix
of residuals that can be related to impact on the environment. This in turn
determines the management strategy for meeting air and water quality standards
as well as socioeconomic criteria.
Controlled field exposures to selected pollutants, such as those being
conducted by the CFPPP research team (described subsequently) can enable
management to determine the exact relationship between the rate of exposure to
a particular pollutant and the response of local biota. Such prediction of
specific environmental impacts at a specific site could form the basis for more
rational siting decisions (Woodwell, 1970).
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ECOLOGICAL CONCEPTUAL FRAMEWORK
A large and expanding body of theory and empirical information dealing with
stress ecology serves as the basis for an ecological model of the effects of
coal-fired power plant emissions on northern grassland ecosystems. A
conceptualization of this is shown in Figure 1. The "target population" may be
defined either taxonomically or functionally. An ecosystem functional model can
be constructed by coupling the models of functionally important "target
populations." The degree of resolution within the ecosystem model can be set by
the definitions assigned to "target populations." By examining the response of
the model to changes in air quality over time it is possible to characterize the
"emergent" properties of the system.
The quality and magnitude of direct effects of pollutants on organismic
function will undoubtedly vary among species and all higher taxonomic groups.
This differential selective pressure provides the basis for indirect effects
upon less sensitive biota and upon ecosystem structure and dynamics.
Investigators in this project are interested in the relative sensitivities of
species to both direct and indirect effects and the relative homeostatic and
competitive capabilities of these species. The time lag (which may or may not
be appreciable) between the occurrence of direct effects and the appearance of
indirect effects and the degree of reversibility of these effects are also of
interest.
A number of parameters and functions should be investigated as potential
progenitors of systems-level indirect effects in northern plains grasslands.
Effects on critical resources such as soil moisture, nutrients, solar radiation,
and trace elements are likely to be very important.
Much of the dynamics of the northern plains grassland ecosystem appears to
be regulated/controlled by the vagaries of the water cycle (Van Dyne, 1973).
Particulate emissions can affect the formation and building of clouds by acting
as cloud condensation nuclei or ice nuclei upon which water vapor condenses.
The hot, dry air that accompanies these aerosols as they are swept into the base
of clouds may also be important. Under certain conditions precipitation may be
increased by the presence of particulate emissions. Under other circumstances
precipitation may be reduced (Library of Congress, 1975). This could alter the
amount of water to biological components of the ecosystem and could lead to
major shifts in successional trends. Water stress may interact synergistical ly
with potentially toxic air emissions, affecting the susceptibility of some
system components to direct toxic effects.
By influencing the degree and nature of cloud formation, and by direct
scattering and adsorption, particulate emissions may affect the quality and
quantity of solar radiation impinging on the ecosystem. This could influence
primary productivity selectively, by species, and thus serve as a major
modifying force.
Certain gaseous emissions may influence the nutrient cycle. At low levels,
S02 and NO may serve as a source of plant nutrients. If portions of the system
are nutrient-limited, these low level emissions could have a stimulatory effect
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MULTIDIMENSIONAL ARRAY OF STACK EFFLUENTS
DIRECT (TOXIC)
EFFECTS
FOOD CHAIN
CONCENTRATION
DIRECT (TOXIC) EFFECTS
INDIRECT EFFECTS
VIA
ALTERED PRIMARY
PRODUCTION
MULTIDIMENSIONAL
PREDATOR POPULATIONS
adjustment
of
pred at ion
INDIRECT EFFECTS
VIA
ALTERED ENVIRONMENTAL
QUALITY
MULTIDIMENSIONAL
TARGET POPULATION
adjustment
of competitive
balance
adjustment of
life table parameters
T
MULTIDIMENSIONAL
COMPETITOR POPULATIONS
EFFECTS ON
TARGET POPULATION
Figure ]. Autecological perspective of pollutant-ecosystem interaction.
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on plant growth. At higher levels, such emissions begin to have direct toxic
effects which may override the stimulatory effects of low level emissions.
When primary critical resources are affected, changes in physiological
functions or population parameters of "key" species will quickly follow. These
may include changes in growth rate, maturation time, reproductive rate, and rate
of phenological development. Changes in populations that are of sufficient
magnitude will alter community structure. Proportions of species may begin to
change, new species may successfully invade. Of interest is how these
structural changes affect community productivity and stability, and also the
degree of reversibility of these changes.
APPLICATION OF PREDICTIVE ASSESSMENT
There are nearly as many approaches to the selection of power plant sites
as there are utilities in this country. The process is governed by many
individual circumstances and a diversity of local and state laws and procedures.
Further, the business concerns of the power company or utilities involved, value
judgments by various decision makers, legislation, lobbies and the potential for
adversary proceedings can influence all stages of the decision process.
In general, control of the location of power plants and attendant
facilities resides with state governments. Policies differ among states and in
some, comprehensive siting laws have been enacted. Montana, for example, has a
planning mechanism that requires state certification. Plans for specific energy
facilities must be submitted to the state ten years in advance of construction.
The state evaluates the collective prospective impact of all facilities and
certifies individual plants on the basis of this review. In many states,
however, actual zoning, review, and certification of power plants is delegated
to local governments; state review may not even be required. The individual
company usually selects a site and seeks whatever zoning changes may be
required.
The selection of sites for new plants appears often to be haphazard and is
initially dictated by such factors as ownership or availability of lease lands
and by engineering constraints. From an often large set of available sites,
only a few will be subjected to detailed feasibility studies. Perhaps one to
three are then evaluated according to NEPA guidelines to determine whether or
not the proposed facility will have an acceptable level of environmental impact.
The selection of appropriate sites can be very costly, and in recent years, a
number of proposed facilities have been denied permits following the review of
environmental impact statements. Often the utility has no clear idea until this
time whether or not the proposed facility has an opportunity for approval.
The objective of this project is to develop a procedure that when applied
will yield biological effects information and solutions that will contribute to
the site selection decision. For the present, efforts are restricted to
consideration of the biological effects of coal-fired power plant emissions on
terrestrial ecosystems of the northern great plains with the Colstrip CFPP
Project providing specific data and the first application of certain protocol
elements. Within these constraints project investigators hope to develop
methods to predict a set of futures and their range of probabilities and to
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utilize this information to influence the course of events so that undesirable
impacts of CFPP operation may be ameliorated.
The prediction of environmental impact, however effective, is of no value
to the siting process unless it is appropriately wedded to the constraints and
needs of the decision making processes. Representatives from government
regulatory agencies, producer and user industries, environmental groups and
citizen groups must be included early in the protocol development process in
order to define the policy and user constraints under which the protocol must be
developed. Continuing consultation should be maintained with these policy
groups throughout the process of protocol development.
ELEMENTS OF A SITING PROTOCOL
The protocol will consist of a methodology to select suitable sites for
construction of coal-fired power plants given several alternatives and a
continuing post-siting methodology to ameliorate undesirable biological effects
should they appear. Post-siting developments and continuing information inputs
to the protocol should favor flexible or adaptive operational management of the
working facilties. The chronology of information and action flow is displayed
in Figure 2.
The annual patterns of concentrations of pollutants that would result from
constructing a coal-fired power plant at each of the alternative sites could be
predicted by a plume dispersion model or plume tracing studies.
A set of widely acceptable methods for gathering "baseline" information on
the structure and dynamics of the terrestrial ecosystems at alternative sites is
also needed. The pollution predictions and "baseline" ecosystems information
will then serve as inputs to one or a series of biological response models whose
outputs predict the biological effects of predicted pollutant concentrations on
the ecosystem under consideration.
This information on potential biological effects on several alternative
sites must be considered in conjunction with outputs from a socioeconomic and
engineering protocol to arrive at a siting decision. In specifying outputs for
the biological response models, provision must be made for interfacing with a
socioeconomic segment. Ecological effects must be translated into predictive
alternatives in man's relationship to the ecosystem (e.g., ahanges in
agricultural productivity, changes in numbers of esthetic species, changes in
number of recreational wildlife species, changes in ecosystem management
practices).
Once a decision to site a plant has been made and construction is
completed^the post-siting study is implemented. Data gathering is now directed
to monitoring pollutant concentrations and biological effects in the vicinity of
the power plant. These data are used to validate and improve the predictive
models employed in the site-selection protocol. They will also provide the
information base for adaptive management of the power plant. If pollution
concentrations or biological effects reach unacceptable levels, government
and/or management will have the necessary information to make rational modifi-
cations of plant operations.
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POLLUTION PREDICTION
(DIFFUSION MODELS)
O
SITE CHARACTERIZATION
(SEVERAL ALTERNATIVES)
O
o
o
BIOEFFECTS RESPONSE
MODELS
OUTPUT FROM OTHER
PROTOCOL ELEMENTS
(SOCIOECONOMIC)
SITE SELECTION
(BEST ALTERNATIVE)
o
I-
o
<
o
MONITOR
POLLUTION
MONITOR BIOLOGICAL
EFFECTS
OUTPUT FROM
SOCIOECONOMIC POST
SITING PROTOCOL
ADAPTIVE
MANAGEMENT
Figure 2. Generalized flow diagram of the ecological effects protocol.
9
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The final bioeffects protocol as presently conceived will consist of five
modules.
1. A site characterization module will include a discussion of preferred
methods in collecting "baseline" data and a discussion of the constraints and
limitations of these methods. This module should be amenable to stepwise
application.
(a) Background Assessments would be conducted on numerous sites with
limited field data supplemented by existing literature.
(b) For a few sites with favorable characteristics, a more rigorous
Selection Assessment would be conducted.
(c) For the site selected, a comprehensive Pre-Construction Assessment
would be conducted.
(d) Finally, a long-term Operations Assessment would be conducted to
verify impact predictions, to determine multi-year ecological effects and
to provide the basis for adaptive management.
2. An analytical module will define data handling and synthesis requirements
and procedures.
3. A predictive module will detail and discuss predictive models that may be
used to estimate future impacts from CFPP operation at alternative sites. This
module should list the site specific parameters that must be measured in order
to use these models and the appropriate methods to be used in gathering the
data. As far as possible, the accuracy and precision of these models should be
specified here.
4. A validation module will describe the data requirements, data collection
methods, and procedures for validating and improving predictive models in the
previous module.
5. An adaptive management module will discuss alternatives that are available
to management to ameliorate impact once the CFPP is in operation. This might
include the use of weather forecasts and coal conversion schedules to predict
pollution levels. Dose-response (or exposure rate) models from the predictive
module could be used to define acceptable pollution levels. Coal conversion
activities could then be regulated to insure that acceptable pollution levels
are not exceeded.
SUMMARY
To date, principal investigators have developed a working knowledge of the
ecology of grasslands in the vicinity of Colstrip, Montana. This knowledge,
together with an innovative field experimental method for the assessment of
ecological impact of air pollution, is slowly providing the information required
to develop predictive capability. Such a capability will allow managers to plan
and to control the siting process, to modulate (or ameliorate) impacts through
adaptive management and to anticipate the possibility and needs for rehabili-
10
-------
tation. Improved management should be effected not only through more effective
and efficient site-selection procedures, but also through evolution of the
siting protocol (update and validation) and the adaptive management of power
facilities through all phases of their life cycle.
REFERENCES
Federal Power Commission. In press. The need for flue gas desulfurization
(FDG): ecological effects of air pollutants.
Lawton, J. H. and S. McNeil. 1973. Primary production and pollution.
Biologist 20:3-11.
Library of Congress. 1975. Effects of chronic exposure to low level pollutants
in the environment. Congressional Research Service, Serial 0, pp. 402.
Van Dyne, G. M. 1973. Analysis of structure, function and utilization of
grassland ecosystems. Natural Resource Ecology Laboratory, Colorado State
University, Fort Collins.
Woodwell, G. M. 1970. Effects of pollution on the structure and physiology of
ecosystems. Science, 168:429-433.
11
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COLSTRIP STUDIES
12
-------
SECTION 1
EFFECTS OF S02 AND OTHER COAL-FIRING PLANT
EMISSIONS ON PRODUCER, INVERTEBRATE CONSUMER,
AND DECOMPOSER STRUCTURE AND FUNCTION IN
THE VICINITY OF COLSTRIP, MONTANA
by
W. K. Lauenroth, J. L. Dodd, R. K. Heitschmidt and J. W. Leetham
INTRODUCTION
Within the next 25 years the northern Great Plains of the United States
will be subjected to air pollution from energy development. The air pollution
will come from the conversion of coal to electricity through steam generating
power plants. Most of the coal in the northern Great Plains lies under native
grassland and much of it will be converted to electricity near the open mines.
Sulfur dioxide and other air pollutants from the conversion process will be
released onto the adjacent native grasslands.
The objectives of this study are to look at the effect of air pollution on
primary producers, arthropod consumers, and decomposition processes. The study
has three aspects. At Colstrip, this consists of baseline monitoring of
decomposition processes and productivity and seasonal biomass dynamics of
primary producers and arthropods at four different sites that range from 11 to
18 km east and southeast of the power plant. Baseline monitoring began in 1974
and will continue over the next several years to determine what, if any,
influence air pollution has on these grasslands.
COLSTRIP MONITORING SITES
The 1975 growing season marked the end of a baseline data collection phase
for the Colstrip Power Plant Project. Unit I of the generating complex at
Colstrip began operation in the fall of 1975 and the growing season of 1976
represents the first year of data collection under the influence of the power
plant operation. An important task now to be completed with respect to the
baseline data is to compare and contrast the two years of baseline data
collection, 1974 and 1975, with respect to abiotic conditions in an attempt to
relate differences in ecosystem response to differences in abiotic conditions
during the two years. The objective of this report is to present the 1975 data
and make some initial comparisons with the data collected in 1974.
13
-------
ABIOTIC DATA
The collection of abiotic data at the Colstrip sites was not begun until
the growing season of 1975. The U.S. Department of Commerce weather data for
southeastern Montana can be used to make general comparisons between the two
years. Precipitation data for 1974 and 1975 are recorded in Table 1.1. Data for
1974 are from the U.S. Department of Commerce Weather Station at Colstrip and
1975 data were collected at the four monitoring sites. The Colstrip Station
(USDC) apparently was not operated in 1975 and data are not available to make
comparisons between 1974 and 1975 to see how these data compare with data
collected at the monitoring sites. Total precipitation for the May to September
period was 228 mm in 1974 and 261 mm average for the four sites in 1975. More
important than the differences in total precipitation were the differences in
the May and June precipitation. In 1974, 121 mm of precipitation were recorded
during May and June and in 1975, 221 mm were measured.
The average monthly temperatures in Table 1.2 are presented for general
comparison of the growing seasons during 1974 and 1975 and do not necessarily
represent average monthly temperatures for the Colstrip sites. The major
differences between the two years included a much cooler April, a slightly
warmer May, and a cooler June.
Soil water data collected at the Colstrip sites in 1975 reflected the high
precipitation during May and June (Table 1.3). Soil water to a depth of 105 cm
remained above 20 cm on all sites through 30 June. This was followed by a rapid
depletion during the first two weeks of July and a subsequent gradual depletion
throughout the remainder of the growing season. No major differences were
evident among sites except for small differences in total soil water observed
throughout the period. The Hay Coulee and Kluver East sites apparently have the
highest water-holding capacity and the Kluver North site the lowest. Soil water
at the end of the growing season was highest at the Kluver East site and lowest
at the Kluver North site and averaged approximately 10 cm to a depth of 105 cm
for al1 sites.
Soil surveys of the Colstrip sites were made in 1975. Although physical
and chemical descriptions are incomplete at this time, profile descriptions for
Hay Coulee, Kluver West, and Kluver East are included in the Appendix.
PLANT PHENOLOGY
Phenological observations were made on a weekly basis using a 14-stage
phenological classification (Table 1.4) during the 1975 season. Fourteen
species were observed and represented a range from Hood's phlox which was
flowering at the time of the first observation to fringed sagewort which had
just begun flowering at the time of the last observation in mid-September
(Tables 1.5 and 1.6). Few differences in the timing of phenological events were
among sites with stage 14, drought-induced dormancy, being the most variable of
the stages and presumably reflecting differences in soil water among sites.
14
-------
TABLE 1.1. GROWING SEASON PRECIPITATION (mm) FOR THE COLSTRIP SITES DURING
THE GROWING SEASON IN 1974 AND 1975.
Colstrip sites 1975
Month
May
June
July
Aug.
Sept.
Total
Col strip
1974
94
27
50
28
29
228
Hay Coulee
121
94
17
16
5
253
Kluver West Kluver North
121 120
93 116
23 13
21 19
5 5
263 273
Kluver East Average
113 119
104 102
14 17
15 18
6 5
252 261
TABLE 1.2. AVERAGE MONTHLY TEMPERATURE (°F) FOR SOUTHERN MONTANA DURING 1974
AND 1975.
Month
April
May
June
July
Aug.
Sept.
Southeastern
1974
46.5
50.8
65.2
73.9
63.7
55.0
Montana average
1975
38.4
52.5
61.3
73.0
67.1
56.2
15
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TABLE 1.3. SEASONAL DYNAMICS OF SOIL WATER (cm), COLSTRIP STUDY SITES, 1975.
Depth
Treatment (cm)
Hay
Coulee
TOTAL
Kl uver
West
TOTAL
Kluver
North
TOTAL
Kl uver
East
TOTAL
0-15
15-30
30-45
45-60
60-75
75-90
90-105
0-15
15-30
30-45
45-60
60-75
75-90
90-105
0-15
15-30
30-45
45-60
60-75
75-90
90-105
0-15
15-30
30-45
45-60
60-75
75-90
90-105
3 May
3.73 + .
4.04 + .
4.54 + .
4.79 + .
4.17 + .
3.34 + .
1.73 + .
26.35
3.36 + .
3.34 + .
3.77 + .
3.90 + .
3.31 i .
3.70 + .
3.25 + .
24.63
3.04 + .
3.39 + .
3.83 + .
4.72 + .
3.63 +
2.91 + .
2.42 + .
23.93
3.52 + .
19
37
06
47
31
35
20
11
05
03
16
08
29
33
20
20
20
92
14
34
14
07
4.16 + .42
4.36 + .
4.05 + .
3.99 + .
3.51 + .
2.01 + .
25.61
27
11
19
48
71
14 May
2.95 +
4.00 +
.24
.28
4.39 + .34
4.29 +
3.68 +
3.43 +
3.30 +
26.
2.96 +
3.27 +
3.01 +
2.83 +
3.39 +
4.23 +
4.07 +
23.
2.30 +
2.41 +
3.47 +
3.46 +
3.56 +
3.66 +
3.16 +
22.
2.97 +
3.61 +
4.15 +
4.^7 +
3.99 +
4.35 +
4.11 +
27.
.36
.63
46
.26
04
.29
.03
.56
.78
.18
.20
.25
77
.03
.63
.07
.13
.09
.26
.23
03
.26
.69
.32
.23
.25
.16
.28
24
4 June
1 .95 + .
2.33 + .
3.52 + .
3.97 + .
3.20 + .
3.09 + .
3.16 + .
21.22
1 . 98 + .
2.78 + .
3.12 + .
2.87 + .
3.23 + .
3.02 + .
3.06 + .
20.07
1.93 + .
2.50 + .
3.13 + .
5.33 + 1
2.97 + .
3.50 + .
3.60 + .
22.96
2.21 + .
3.57 + .
3.22 +
3.25 + .
3.14 + .
3.65 + .
3.37 + .
22.42
10
55
34
04
15
50
35
19
23
20
15
52
40
39
06
31
39
.80
53
31
30
13
33
19
17
65
08
28
19 June
4.13 + .
3.54 +
3.36 + .
3.28 + .
3.44 + .
3.45 + .
3.28 + .
24.49
2.03 + .
2.70 - .
2.56 + .
3.46 + .
3.44 + .
2.76 + .
3.54 + .
20.49
2.28 3 .
3.16 + .
3.25 + .
3.44 + .
3.30 + .
3.26 + .
3.43 + .
22.12
2.76 + .
3.46 +
3.15 + .
3.47 + .
3.42 + .
3.37 + .
3.44 + .
23.07
18
49
32
33
15
39
55
07
20
08
61
09
23
63
13
20
19
43
46
38
46
21
40
63
10
27
33
32
30 June
1.99 + .
2.70 + .
3.23 + .
3.19 + .
2.95 + .
3.21 + .
2.93 + .
20.20
2.23 +
2.78 ^ .
3.16 +
3.31 + .
3.14 + .
3.29 + .
3.40 + .
21.31
2.40 + .
2.99 + .
3.15 + .
3.57 + .
3.44 + .
3.46 + .
3.42 + .
22.43
2.26 + .
2.66 + .
2.87 + .
3.02 + .
3.43 + .
3.14 + .
2.96 + .
20.36
21
41
30
33
22
40
34
19
78
44
51
42
36
64
50
47
08
03
18
27
12
16
31
16
07
66
31
09
14 July
0.99 + .
1 .42 + .
1.83 + .
2.03 + .
2.22 + .
2.32 + .
2.53 + .
13.43
0.72 + .
1.21 3 .
1 .50 + .
1 .67 + .
2.64 + .
2.67 + .
2.49 + .
12.91
1.19 + .
1 .61 + .
2.00 + .
2.55 + .
2.54 + .
2.54 + .
2.93 + .
15.36
1.22 + .
1.81 +.
2.14 + .
2.19 + .
2.60 + .
2.64 + .
2.66 + .
15.26
29 July
09
22
18
18
28
40
45
04
08
04
13
82
77
53
19
26
21
40
54
55
42
16
30
09
22
21
26
20
1.33 +
1.55 +
1.92 +
2.44 +
2.67 +
2.88 +
2.23 +
15.
0.96 +
1.51 +
2.17 +
2.53 +
2.56 +
2.40 +
2.60 +
14.
0.94 +
1 .44 +
1.67 +
1.93 +
2.05 +
2.36 +
2.70 +
13.
0.88 +
1.34 +
1.39 +
1.60 +
1.92 +
2.66 +
2.94 +
12.
.09
.18
.12
.16
.49
.66
.12
01
.13
30
.31
.23
.12
.34
.43
73
.08
.20
.14
.27
.27
.16
.26
09
16
.22
17
13
.35
.29
.34
74
9 August
0.80 + .
1.09 + .
1.44 + .
1 .96 + .
2.27 + .
2.72 +
2.02 + .
12.30
0.59 + .
1 . 00 + .
1.07 + .
1.14 + .
1.50 + .
1 . 84 + .
2.13 +
9.27
0.60 + .
0.94 + .
1.06 + .
1.24 + .
1.20 + .
1 .53 + .
2.09 + .
14
25
26
84
36
16
18
04
13
13
07
37
64
18
03
10
11
11
10
19
37
8.66
0.99 + .
1.16 + .
1.19 + .
1.69 + .
2.07 + .
2.29 + .
2.90 + .
12.29
37
12
11
26
09
21
47
25 August
0.59 + .
1.10+ .
1.51 + .
1.56 + .
1 .80 + .
1.85 + .
2.07 + .
10.49
0.73 + .
0.95 + .
0.94 + .
1.10 + .
1.45 + .
2.70 + .
2.38 + .
10.25
0.73 + .
1.13 + .
1.24 + .
1 .67 + .
1.63 + .
1.83 + .
2.21 + .
04
23
25
12
19
21
27
11
09
01
13
18
22
75
09
06
07
31
36
56
52
9 September
0.69
1.21
1.65
1.41
1.84
1.53
2.15
10
0.68
1.28
1.02
1.57
1.74
2.45
1.81
10.
0.63
1.12
1 .30
1.01
1 .32
1.55
1.81
10.43 £
0.78 + .
1.12 + .
1.77 +.
1.28 + .
1.84 + .
2.35 + .
2.47 + .
11.61
11
11
66
06
17
36
51
0.67
1.16
1.35
1.27
1.91
2.28
2.62
11
+ .05
+ .19
+ .46
+ .24
+ .33
+ .14
+ .33
.48
+ .04
+ .37
+ .10
+ .41
+ .54
+ .77
+ .35
56
+ .13
+ .25
+ .21
+ .25
+ .30
+ .47
+ .64
1.73
+ .15
+ .10
+ .12
+ .09
+ .13
+ .06
+ .34
.26
-------
TABLE 1.4. PHENOLOGICAL STAGES USED DURING THE 1975 GROWING SEASON.
Phenological stages
1.. Winter dormancy
2. First visible growth
3. First leaves fully expanded
4. Middle leaves fully visible
5. Middle leaves fully expanded
6. Late leaves fully expanded
7. First floral buds
8. Mature floral buds
9. Floral buds and open flowers
10. Floral buds, open flowers, and ripening fruit
11. Buds, flowers, and green and ripe fruit
12. Buds, flowers, green and ripe fruit, and seeds
13. Green and ripe fruit, and dispersing seeds
14. Dispersing seeds and drought-induced dormancy
17
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TABLE 1.5. PHENOLOGY OF MAJOR FORB SPECIES, COLSTRIP STUDY SITES, 1975 (A = Hay Coulee, B = Kluver West, C = Kluver North, D = Kluver East).
oc
Phlox Tragopoqon Hedeoma Sphaeralcea
hoodji dubius h i s p i da cocci nea
Date
27 May
2 June
11 June
18 June
23 June
30 June
8 July
14 July
22 July
28 July
3 Aug.
10 Aug.
18 Aug.
26 Aug.
1 Sept.
8 Sept.
14 Sept.
ABCD ABCD ABCD A R
11 11 11 11 55554 4
13 13 13 12 7777 4 5
14 14 13 877 4 5
14 9 8 9 9 8 7
14 9 9 11 10 9 8
11 10 11 10 10 10 10 10 10 9
13 13 13 13 12 12 12 10 11
13 13 13 13 13 13 13 12 13 11
13 14 14 14 13 13 13 13 13 11
13 14 14 14 14 13 13
14 13 13
13
14 14
C D
4
5
5
9
10
10
10
12
12
13
13
14
Plantaqo Psora lea
Patagonia araophvlla
ABCD ABC
2 3
2 43
344
7 7
8 7
10 9 9 9 83
10 10 10 9 10 10
10 10 10 10 10 10
13 13 12 12 10 10
13 13 13 13 10 10
13 13 13 13 13 13
14 13 13 13 13 13
14 14 14 13
D
2
3
7
7
8
9
10
10
10
13
13
13
13
14
Artemisia
frig
A
4
4
5
5
5
5
5
5
6
6
6
6
7
9
9
9
B
4
4
4
4
4
4
4
5
6
6
6
6
7
8
8
9
ida
C
4
4
4
4
4
4
4
5
6
6
6
6
7
9
9
9
D
3
4
4
4
4
4
5
5
5
6
6
6
7
9
9
9
-------
TABLE 1.6. PHENOLOGY OF MAJOR GRASS SPECIES, COLSTRIP STUDY SITES, 1975 (A = Hay Coulee, B = Kluver West, C = i'.luver North, D = Kluver East).
Poa secunda Stipa conata
Date ABCD ABCD
27 May
2 June
11 June 9
19 June 9 889
23 June 10 999
30 June 10 10 10 9 999
8 July 12 12 12 11 10 11 11 10
14 July 13 13 13 12 13 13 13 13
22 July 14 14 14 13 14 13 13 13
28 July 13 13 13 13
3 August 14 14 14 14
10 August
18 August
26 August
1 September
8 September
14 September
Bromus
.iaponicus
ABCD
7777
8888
888
9889
9999
9999
10 9 9 10
10 9 10 12
13 12 13 13
13 13 13 13
14 14 14 14
Koeleria
cristata
ABCD
8
8
9 9
9999
9999
9 9 9 11
11 12 12 12
13 13 13
13 13 13 13
13 13 13 13
13 13 13 13
14 14 14 14
Aristi'cla
loncnseta
ABCD
3
4
4
7
7
8 8
8 8
8 8
10 10
n 10
13 13
13 13
13 14
14
Bouteloua
gracilis
A
4
4
4
4
5
5
7
9
9
10
11
12
13
B
4
4
4
4
4
5
6
6
8
10
11
12
13
13
14
C
4
4
4
4
4
5
6
8
9
10
11
12
13
13
14
D
4
4
5
5
5
7
8
9
10
n
12
13
Aqropyron
smithii
A
4
4
4
6
9
9
10
10
10
11
11
11
12
13
13
13
B
4
4
4
5
6
8
9
9
9
10
n
n
12
13
13
13
C
4
4
4
6
7
9
9
10
10
n
n
12
12
13
13
13
D
4
4
7
7
8
9
10
10
n
n
n
12
13
13
13
-------
ABOVEGROUND BIOMASS DYNAMICS
Figure 1.1 presents seasonal dynamics of total aboveground biomass for the
four Colstrip sites during 1974 and 1975. Represented here are current live,
recent dead (material that died during growing season of measurement) and old
dead (material that died prior to growing season of measurement) biomass. The
immediate differences observable on this figure are a larger amount of current
live biomass in 1975 and the slight delay in the recording of the peak in current
live biomass. Another difference between years, although not an unexpected one,
is the larger amount of old dead biomass recorded in the spring of 1975 compared
to 1974. This is a direct result of the exclusion of cattle by fencing and
points to a problem that must be addressed in analyzing the baseline data. We
must be able to sort out differences between the two years both in terms of
differences in abiotic regimes and differences resulting from fencing. The
Kluver East site which showed the largest increase in current live biomass
between 1974 and 1975 also had the largest response to fencing. Although we
have not looked at the data in terms of species composition, we expect that the
effects of fencing on this site will be larger than on the remaining sites.
Figure 1.2 compares the current season production of cool season grasses,
the most important functional group on the study sites, between 1974 and 1975.
Again, the major differences are the larger absolute amount of current season
production during 1975 and the slight delay in peak production. In 1974, peak
production was recorded on approximately 15 July 1975. Although there was an
increase in the production of cool season grasses in 1975, the relative position
of each site remained constant for both years. The greatest amount of cool
season grasses was found on the Kluver West site in both years and the least
amount of cool season grass production was recorded for the Kluver North site
for both seasons.
Figures 1.3 and 1.4 present current season's production for western
wheatgrass and needle-and-thread grass, the two major contributors to the cool
season grass group. Similar trends of greater production can be seen for both
species during 1975 than during 1974. Western wheat-grass showed much less
fluctuation in the measurement of current season's production from date to date
in 1975 than in 1974. Needle-and-thread grass productivity was the greatest on
the Kluver West site and not measurable for the Hay Coulee site. The Kluver East
site had the least amount of needle-and-thread grass productivity in both 1974
and 1975.
Cool season forb productivity averaged approximately 10 g/m2 for all sites
in 1974 (Figure 1.5). It was only slightly higher (approximately 15 g/m2) in
1975. The Kluver North site had the largest productivity of cool season forbs
in both years and the Kluver East site the least amount of cool season forb
production.
Half-shrub productivity was also greater in 1975 than in 1974 (Figure 1.6).
The abrupt drop in half-shrub biomass recorded at the beginning of July in 1974
was not present in 1975. The Kluver East site had the greatest productivity of
half-shrubs in 1975 while the Kluver North site had the greatest productivity in
1974. This again is an indication of the changes that are occurring in the
Kluver East site, presumably as a result of fencing.
20
-------
o
00
150-
100-
Liwt
— R*c*nt d«od
— Old dead
I5O-
100-
50-
o4r*
APRIL MAY
^&Tt~a asTeM & is 25 s is as
JUNE JULY AUGUST SEPTEMBER APRIL MAY JUNE JULY AUGUST SEPTEMBER
Figure 1.1. Comparison of aboveground biomass dynamics during the 1974 and 1975
growing seasons at the four Colstrip sites: (a) Hay Coulee, (b) Kluver
West, (c) Kluver North, and (d) Kluver East.
-------
100
50
-------
100
Kluver East
50
OJ
c/)
en
o
CD
0
100
50
— Hay Coulee
---Kluver West
— Kluver North
a
\
15 25 5 15 25
APRIL
MAY
JUNE
JULY
5 15 25
AUGUST SEPTEMBER
Figure 1.3, Production of western wheatgrass at the Colstrip sites during
the 1974 (b) and 1975 (a) growing seasons.
23
-------
CJ
GO
CO
o
CD
100 - —
50
0
100
Kluver West
Kluver North
— Kluver East
a
50
0
-H—~
J L
5 15 25 5 15 25 5 15 25 5 15 25 5 15 25
MAY JUNE JULY AUGUST SEPTEMBER
Figure 1.4. Production of needle-and-thread grass at the Colstrip sites
during the 1974 (b) and 1975 (a) growing seasons.
24
-------
100
Hay Coulee
•Kluver West
Kluver North
Kluver East
a
50
CvJ
co
CO
o
CD
0
100
50
5 15 25 5 15 25 5 15 25
APRIL
MAY
JUNE
JULY
AUGUST SEPTEMBER
Figure 1.5. Production of cool season forbs at the Colstrip sites during
the 1974 (b) and 1975 (a) growing seasons.
25
-------
100
50
• Hay Coulee
Kluver North
Kluver East
a
3 100
s
o
GO
50
0
5 15 25 5 15 25 5 15 25 5 15 25 5 15 25
MAY JUNE JULY AUGUST SEPTEMBER
Figure 1.6. Production of half-shrubs at the Colstrip sites during the
1974 (b) and 1975 (a) growing seasons.
26
-------
Warm season grasses are generally an unimportant component in terms of
total productivity of these sites and very small differences among the two years
were recorded (Figure 1.7). The Hay Coulee site had the greatest productivity
of warm season grasses in both years and was approximately 15 g/m2. Slight
differences were seen for the remaining sites with an increase in warm season
grasses for both the Kluver West and Kluver East sites.
Aboveground net primary production (ANPP) was calculated by summing the
peak values of current live plus recent dead biomass for each functional group.
Figure 1.8 shows ANPP for the four sites for 1974 and 1975 with the contribution
of each functional group. In 1974 ANPP estimates ranged from 106 g/m2/yr for
the Kluver East site to 123 g/m2/yr for the Kluver North site. The ANPP in 1975
was greater for all sites, ranging from 131 g/m2/yr for Kluver West to 165
g/m2/yr for Kluver East. The percentage contribution of the functional groups
to ANPP was similar between years for all sites with the maximum differences on
the order of 5% to 7%. An example of this is the data from the Kluver North
site. Here the percentage contribution of half-shrubs increased 5% from 1974 to
1975, the same for cool season forbs decreased 7%, warm season grasses increased
5%, and cool season grasses decreased 4%. The explanation for these differences
is related to the interaction of exclusion from grazing and increased
precipitation in 1975. At this time we are not able to separate the effect of
these influences.
BELOWGROUND PRODUCER BIOMASS
The seasonal dynamics data for belowground biomass require further
analyses and are not discussed in this report. Analyses of the depth
distribution of root biomass are complete. Figure 1.9 presents the vertical
distribution of root biomass for six sites over 2 years. Although there is some
variability in the depth distribution of the root systems of these northern
mixed prairie grasslands, on the average they are quite similar. Averaging over
all sites in both years approximately 51% of the total root biomass is found in
the 0-10 cm depth. This value decreases to 4% of the total root biomass found in
the 50-60 cm depth. These averages are similar to root distributions reported
by other researchers (Dodd et al_. , 1974; Bartos and Sims, 1974).
LITTER DYNAMICS
Changes in litter standing crop on the Colstrip sites for 1974 and 1975
indicate general similarity between sites and years (Table 1.7). Although
several dissimilarities among sites are apparent for the 1974 season and the
early portion of the 1975 season, the variation between sites by the end of 1975
(August and September) was small. The early differences are attributed to lag
effects from differential grazing histories prior to 1974; the authors conclude
that, with respect to litter production and decomposition, the sites are now
comparable and will likely remain so unless differences are induced by site-to-
site variations in weather or air pollution.
ARTHROPODS
The arthropod fauna of the four EPA field sites near Colstrip, Montana (Hay
Coulee, Kluver West, Kluver North, and Kluver East) were sampled by three
27
-------
100-
50
-------
150-
Season Grasses
Season Grasses
Cool Season Forbs
Warm Season Forbs
Half Shrubs
a
OJ
o
i-
o
=)
a
o
a:
o.
ct:
LU
UJ
100
50
o
50
100
50
0
Figure 1.8.
\\\\\\\\\\\N
Hay Coulee
Kluver West
Kluver North
Kluver East
Total aboveground net pv^imary production for the Colstrip
sites during 1974 (a) and 1975 (b).
29
-------
0-10
10-20
20-30
E
o
30-40
LiJ
Q
40-50
50-60 -
•Colstrip - 1974
•Colstrip - 1975
Ash Creek - 1974
Taylor Creek -1975
VERTICAL ROOT DISTRIBUTION
Depth
0-10
10 -20
20 -30
30 -40
40 -50
50 -60
Average
51
16
13
9
7
4
_L
10
20
_L
40
50
60
Figure 1.9.
30
PERCENT
Distribution of root biomass by depth increments for three northern mixed-grass
prairie sites.
-------
TABLE 1.7. LITTER STANDING CROP (X ± SE, ash free g/m2) FOR COLSTRIP SITES,
1974 AND 1975.
Hay Coulee
Date
•974
11 May
11 June
29 June
26 July
16 August
26 September
X
1975
22 April
18 May
20 June
15 July
1 1 August
16 September
X
X
161
172
201
172
128
192
171
162
170
179
154
231
218
186
SE
17
11
13
12
12
15
10
14
11
15
26
18
Kl uver
X
177
175
183
140
139
167
163
159
166
160
207
191
177
West
SE
13
12
6
12
19
13
9
20
10
15
14
Kluver
X
234
157
169
171
198
230
193
193
205
163
218
277
189
207
North
SE
21
22
15
18
10
24
22
19
16
32
33
16
Kl uver
X
158
154
171
161
153
148
157
144
157
152
204
206
144
East
SE
16
12
19
18
18
13
17
19
14
16
39
31
-------
different techniques, each designed for certain components of the total fauna.
For the "aboveground arthropods," i.e., those occurring in the litter and
standing vegetation, a quick trap system was used. An open-bottomed cage of
known size was dropped over preselected, randomly located plots. The cage or
quick trap was dropped from the end of an 18 ft boom mounted on a two-wheeled
cart. The insects, vegetation, and surface litter were removed from the trap by
a vacuum system and the arthropods extracted from the refuse by use of Berlese-
or TulIgren-type extraction.
The "soil macroarthropods," i.e., those arthropods occurring below the
surface litter and large enough to be sieved from the soil with a 1 mm opening
sieve, were sampled by taking 12.5 cm diameter by 15 cm deep soil cores and wet
sieving them. All material retained by the sieve was further separated by
flotation in a saturated solution of magnesium sulfate (MgS04).
The "soil microarthropods," i.e., those occurring in the litter and soil
and small enough to pass through a 1 mm opening sieve, were sampled by taking 5.0
cm diameter by 10.0 cm deep soil cores in such a way as to retain, as nearly as
possible, the original soil structure. The arthropods were extracted by a
Macfadyen high temperature gradient system. Leetham (1975) gives a detailed
discussion of all the above techniques.
In the field at each randomly chosen sample point, one sample of each of
the above types was taken with five such sample points being selected in each of
two replicates of each treatment, i.e., field site. All arthropods collected
were identified to family (genus and species where possible). Representatives
of each group or category were dried at 65° C for 25 hr and weighed to obtain dry
weight biomass data. There were five to six sample dates in each of the 1974 and
1975 seasons.
Because of some recently detected problems in the computer summarizations
of the soil microarthropod data, a discussion of that portion of the arthropod
fauna will be included in a later report.
Tables 1.8 and 1.9 give summarizations of the aboveground and soil macro-
arthropods by total counts (Table 1.8) and by the various trophic or functional
groups (Table 1.9). From Table 1.8 it is apparent that generally there was a
substantial difference between the two seasons with the 1975 season producing
larger numbers of both aboveground arthropod biomass which shows a substantial
reduction, possibly due to a shift in the dominant species between the two
years. There appears to be considerable treatment or field site differences,
however, the significance of these differences may be artificial due to the high
variability of the data. Generally, the Kluver North (C) plot supported the
highest arthropod populations in 1974 but that dominance changed in 1975. Since
no statistical analyses have been made on the data, any significant treatment
and/or year effects are unknown.
Table 1.9 gives a summary of the arthropod fauna by trophic group. Plant
feeding types are all grouped together here rather than splitting them by their
feeding types, i.e, chewing, sucking, pollen feeding, etc. The plant feeding
and omnivorous types appear to be the dominant feeding types, both by numbers
and biomass for both above and belowground arthropods. Within the treatments,
32
-------
TABLE 1.8. SUMMARY, BY TOTAL COUNT, OF THE ARTHROPOD CENSUSING AT THE FOUR
FIELD SITES NEAR COLSTRIP, MONTANA FOR THE 1974 AND 1975 SEASONS
(A = Hay Coulee, B = Kluver West, C = Kluver North, D = Kluver
East).
Group
?1/ 21/
Numbers/m Biomass g/m
ABCDABCD
Aboveqround arthropods
1974 45.2 39.8 96.5 48.3 .143 .269 .345 .200
1975 136.1 99.3 122.2 109.2 .119 .174 .126 .082
Soil macroarthropods
1974
1975
37.
82.
8
2
49.
148.
5
3
74.7
62.5
42.
69.
7
5
.148
.574
.065
.551
.080
.620
.045
.356
-Time weighted means.
33
-------
TABLE 1.9. SUMMARIZATION BY TROPHIC OR FUNCTIONAL GROUP OF THE ARTHROPOD
DATA GIVEN IN TABLE 1.8 (A = Hay Coulee, B = Kluver West, C =
Kluver North, D = Kluver East).
2^
Numbers/m
Group
Aboveground arthropods
1974
Soil
1975
macroarthropods
1974
1975
Aboveground arthropods
1974
Soil
1975
macroarthropods
1974
1975
Aboveground arthropods
1974
Soil
1975
macroarthropods
1974
1975
Aboveground arthropods
1974
1975
A
2.9
11.5
23.2
30. 1
28. 1
49.0
5.8
20.9
6.6
17.1
6.2
11.6
0.5
1.0
B
1
15
3
75
C
D
Unknown
.4 2.2 5.8
.7
.9
.2
Plant
21.6
49
8
18
5
8
1
22
0
0
.1
.9
.8
20.
9.
3.
2 18.4
8 6.3
5 19.6
Feeding
60.6 22.6
63.
9.
25.
0 50.4
7 16.4
1 24.8
Predators
.4 8.9 7.6
.9
.4
.5
13.
9.
17.
7 15.3
9
6 6.9
Parasite id
.4 0.8 0.6
.6
1.
4 0.4
A
.001
.002
.091
.072
.132
.085
.023
.332
.005
.008
.023
.064
>.001
.001
21/
Biomass g/m
B
>.001
.001
.016
.013
.249
.150
.027
.112
.005
.005
>.001
.125
>.001
>.001
C
.001
.001
.024
.004
.326
.100
.032
.410
.005
.012
.002
.127
.001
.001
D
.002
.001
.001
.018
.186
.065
.040
.263
.005
.003
--
.043
.001
>.001
(continued)
34
-------
TABLE 1.9. (continued)
Numbers/m^
Biomass g/m^
Group
B
Soil macroarthropods
1974
1975
Aboveground arthropods
1974 5.0
1975 21.8
Soil macroarthropods
1974 3.0
1975 30.1
Aboveground arthropods
1974 2.1
1975 33.8
Soil macroarthropods
1974
1975 1.7
Aboveground arthropods
1974
1975 1.9
Soil macroarthropods
1974 1.8
1975 14.7
2.5
1.0
Omnivore
5.0 20.0 7.2
12.7 16.0 13.6
25.9 23.7 7.0
75.2 3.5 19.6
Scavenger
5.9 3.3 4.3
13.5 7.7 11.2
5.8 2.9 3.0
6.9 3.8 4.5
Non-feeding
0.1 0.1 0.1
2.9 0.4
2.4 16.8 11.4
23.2 8.5 10.5
.008 .005
.001 .001 .004 .002
.014 .009 .010 .011
.002 .007 .005 .001
.072 .103 .004 .018
.003 .013 .005 .004
.003 .004 .005 .002
.004 .003 .003
.006 .019 .020 .023
>.001 >.001 >.001
,006 .009 .001
,010 .004 .009 .006
,094 .063 .035 .009
-Time weighted means.
35
-------
Kluver North supports the highest numbers and biomass of plant feeders and the
lowest number and biomass of scavengers. The switch in the relationship of
numbers to biomass for aboveground plant feeders, i.e. , lower biomass for higher
numbers in 1975, appears to reflect a change in the dominant species types, as
mentioned earlier. We hope a more in-depth analysis of the data will answer
this question.
Since the data included here have not been statistically analyzed, the
homogeneity of the four field plots is not clearly defined. This is due in part
to the high variability of the data, a common characteristic of this kind of
ecological data. When the statistical analyses are complete, a comparison of
year and date information will provide a more detailed characterization of the
field sites as examples of a northern mixed-grass prairie.
REFERENCES
Bartos, D. L. and P. L. Sims. 1974. Root dynamics of a shortgrass ecosystem.
J. Range Manage. 27:33-36.
Dodd, J. L. , J. K. Lewis, H. L. Hutcheson, and C. L. Hanson. 1974. Abiotic and
herbage dynamics studies at Cottonwood 1971. US/IBP Grassland Biome Tech.
Rep. No. 250. Colorado State University, Fort Collins. 195 pp.
Leetham, J. W. 1975. A summary of field collecting and laboratory processing
equipment and procedures for sampling arthropods at Pawnee Site. US/IBP
Grassland Biome Tech. Rep. No. 284. Colorado State Univ., Fort Collins.
49 p.
36
-------
APPENDIX
SOIL DESCRIPTIONS-7
Soil surveys were conducted at the Kluver East, Kluver West, Kluver North,
and Hay Coulee sites. The morphological properties were examined to a depth of
152 cm in each of two soil excavation pits at each site. Samples were taken from
all horizons in the profiles. Those samples are now being analyzed for physical
and chemical properties.
Preliminary analyses based on morphological properties indicate that the
soils at the Kluver East site belong to Kobar and Lonna series. At the Kluver
West and Kluver North sites the Yamac series predominates, but much of the soil
at the Kluver West site is very stoney and cobbly causing description to be
difficult. The Hay Coulee site has soils of the Yamac and Lonna series.
Representative profile descriptions from these series are presented in profiles
1-3.
Complete analyses and interpretation of these soils await completion of
physical and chemical analyses. Physical properties being determined are
texture, bulk density, and water-holding capacity. Chemical analyses will yield
information for organic matter (carbon) pH, total N, total P, inorganic P,
organic S, total S, inorganic S, cation exchange capacity, exchangeable bases
(Ca, K, Mg, and Na), and lime.
-Soils were described by R. G. Woodmansee with the assistance of Lewis
Daniels, Soil Survey Party Chief, USDA, SCS, Forsythe, Montana.
37
-------
PROFILE 1. Kobar Clay Loam
Kluver East - North Pit
Depth
Horizon (cm) Description
All 0-4 Light brown gray (10 YR 6/2 dry) to dark brown (10
YR 4/3 moist); loam; moderate fine platy; hard when
dry, friable when moist, slightly sticky and
slightly plastic when wet; no lime present; abrupt,
smooth boundary; many fine roots.
A12 4-12 Light alive brown (2.4 YR 5/2 dry) to dark gray
brown (10 YR 4/2 moist); clay loam; weak medium
platy; hard when dry, friable when moist, sticky
and plastic when wet; no lime present; clear,
smooth boundary; many fine roots.
B2 12-18 Pale brown (10 YR 6/3 dry) to dark brown (10 YR 4/3
moist); sandy clay loam; moderate medium prisms
breaking to moderate medium blocks; hard when dry;
friable when moist, sticky and plastic when wet; no
lime present; clear, smooth boundary; many fine
roots.
B3ca 18-36 Very pale brown (10 YR 7/3 dry) to dark brown (10
YR 5/4 moist); sandy clay loam; weak coarse prisms
breaking to moderate medium and fine blocks; hard
when dry, friable when moist, sticky and plastic
when wet; lime present; gradual, smooth boundary;
roots common.
Clca 36-120 Very pale brown (10 YR 7/3 dry) to yellowish brown
(10 YR 5/4 moist); sandy clay loam; massive; hard
when dry, friable when moist, sticky and plastic
when wet; lime present; gradual smooth boundary;
roots common to 45 cm and few roots to 120 cm.
C2ca 120-152 Very pale yellow (10 YR 7/3 dry) to yellowish
brown (10 YR 5/4 moist); clay loam; massive; hard
when dry, friable when moist, sticky and plastic
when wet; lime present; few roots present.
38
-------
PROFILE 2. Lonna Loam
Hay Coulee - North Pit
Depth
Horizon (cm) Description
Al 0-12 Light brown gray (2.5 YR 6/2 dry) to dark brown
crushed (10 YR 4/3 moist) and dark gray brown
coated (10 YR 4/3 moist); loam; weak medium platy;
slightly hard when dry, friable when moist, slightly
sticky, slightly plastic when wet; no lime; clear
smooth boundary; many fine roots.
B2ca 12-22 Pale brown (10 YR 6/3 dry) to yellowish brown
crushed (10 YR 5/4 moist) and dark brown coated (10
YR 4/3 moist); silty clay loam; weak medium prisms
breaking to moderate medium and fine blocks; hard
when dry, friable when moist, sticky and plastic
when wet; lime present; gradual smooth boundary;
many fine roots.
Cl 22-60 Very pale brown (10 YR 7/3 dry) to yellowish brown
ca (10 YR 5/4 dry); silty clay loam; weak, very coarse
platy breaking to moderate fine blocks (plates not
of pedogenic origin); hard when dry, firm when
moist, sticky and plastic when wet; lime present;
gradual smooth boundary; many fine roots to 30 cm,
common fine roots 30-60 cm.
C2 60-98 Very pale brown (10 YR 7/3 dry) to yellowish brown
ca (10 YR 5/4 moist); silty clay loam; massive; hard
when dry, friable when moist, sticky and plastic
when wet; lime present; gradual smooth boundary;
common fine roots to 80 cm, few fine roots to 98
cm.
C3 98-152 Very pale yellow (10 YR 7/3 dry) to light olive
ca brown (2.5 YR 5/4 moist); silty; massive; slightly
hard when dry, friable when moist; lime present;
few roots present.
39
-------
PROFILE 3. Yamac Silt Loam
Kluver West - West Pit
Depth
Horizon (cm) Description
Al 0-10 Brown (10 YR 5/3 dry) to dark gray brown (10 YR
4/2 moist); fine sandy loam; soft medium platy;
soft when dry, very friable when moist, nonsticky,
slightly plastic when wet; no lime; clear, smooth
boundary; many fine roots.
B2 10-31 Yellowish brown (10 YR 5/4 dry) to brown to dark
brown crushed (10 YR 4/3 moist), dry gray brown
coated (10 YR 4/2 moist); sandy clay loam; moderate
medium prisms breaking to moderate medium blocks;
hard when dry, friable when moist, sticky and
plastic when wet; no lime; clear, smooth boundary;
fine roots common.
B3 31-40 Pale brown (10 YR 6/3 dry) to dark gray brown (2.5
YR 4/2 moist); sandy clay loam; weak coarse prisms
breaking to weak coarse and medium blocks; hard
dry, friable when moist, sticky and plastic when
wet; lime present but not visible; gradual smooth
boundary; roots common.
Clca 40-118 Very pale brown (10 YR 7/3 dry), too cobbly and
mixed to determine moist color; cobbly (upper 7
cm); sandy clay loam; massive; hard when dry,
friable when moist, sticky and plastic when moist;
small threads and masses of lime; gradual and wavy
boundary; roots common 40-50 cm, few roots 50-118
cm.
C2ca 118-156 Light gray (2.5 YR 7/2 dry), too variable for moist
color; cobbly sandy loam; massive; too cobbly for
consistence; lime coatings on cobbles; few roots.
Lower horizons of this profile contain unweathered fragments similar to
nearby uplands. Most Yamac soils of EPA study sites do not contain these
cobble layers. The east end of Kluver West contains so much cobble and
gravel that the pit was not described.
40
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SECTION 2
PLANT COMMUNITY STUDIES IN THE VICINITY OF COLSTRIP, MONTANA
by
J. E. Taylor and W. C. Leininger
INTRODUCTION
The plant community monitoring aspects of the Colstrip Coal-fired Power
Plant Project are generally divided into studies of more-or-less quantitative
plant population and community attributes (diversity, phenology; canopy
coverage, productivity) and photographic surveillance, both from ground and
aerial vantage points. This section will deal with diversity, canopy coverage,
and phenology studies.
The procedures and data reported here are interrelated with concurrent
studies of bird and mammal populations (including diversity), plant pathology,
and lichen ecology. In total, all of these studies are contributing to the
development of a generalized, comprehensive approach to air pollution effects
monitoring in biological systems.
The objectives of this aspect of the project are to:
1. Document pre-treatment native plant communities in areas likely to be
affected by the power plants under investigation and on areas to be
stressed artifically with pollutants.
2. Develop measurement techniques and monitor changes in plant community
structure, diversity, phenology, and speciation following air
pol1ution stress.
3. Develop detailed vegetation maps of the study areas.
4. Provide data for simulation models to predict bioenvironmental
changes resulting from fossil fuel power generation in other areas.
The work was initiated on July 15, 1974. Research from previous field
seasons has been reported by Taylor et a_[. (1975 and 1976). The present report
is based on the data collected during the 1975 field season.
This discussion is divided into two principal parts: diversity studies,
using standard indices derived from information theory, and phenology studies,
using an index system of the authors' design.
41
-------
The underlying hypothesis of this work is that, as a plant community is
stressed with the application of any external perturbation, including air
pollution, and physiological changes in members of the community which
result in changed rates of vital functions should be shown by changed species
composition and/or rates of seasonal growth processes. If species composi-
tion is affected, with either loss of sensitive species, or re-ordering of
species dominance in the community, this should be reflected in the species
diversity observed. If rates of normal growth and reproductive functions are
influenced, the phenological profiles of component species should change. In
the case of the Zonal Air Pollution System (ZAPS) plots, where sulfur dioxide
is applied at controlled rates to native vegetation, both of these indications
have been observed (see Section 11).
Both responses to pollution could be studied with biomass/productivity
data, such as those collected by Dodd and associates (see Sections 1 and 10).
However, the authors are interested in developing a non-destructive method-
ology which might be used where biological material was in limited supply, or
where complementary observations could be made, serving to fortify biomass
data by mutual corroboration.
SPECIES DIVERSITY STUDIES
In 1975, 3 sampling times were used, representing (a) "peak of green"
(June 10-11), (b) summer growth and maturity and warm season "peak of green"
(July 10-12), and (c) late season maturity (September 16-17).
Plots within study areas were located by placing a cord with meter-
spaced knots in a random meandering pattern through the sample area. The 2
x 5 dm plot frames were placed at each knot and canopy coverage estimated
using the procedure of Daubenmire (1959). At the same time, species were
counted, the numerical data to go into diversity indices. Previous experi-
mentation had indicated that 2 sets of 20 frames per plot constituted a
statistically adequate sample (i.e., additional observations did not con-
tribute to increased sensitivity of diversity indices).
The diversity index used was the Shannon-Weaver function (Shannon and
Weaver 1949):
*
S
H' = - I Ni_ ln Nj_ Where H1 = Index of diversity
i=l N N S = number of species
Ni = number of ith species
N = total number of all species
The index was calculated in two different ways. In using number data
(plant density), the index was calculated for each of the 50 individual
frames, and the mean of these values was tabulated and graphed. With canopy
coverage data, the index was calculated as an accumulated figure over the
entire set of 40 frames.
42
-------
The first approach allows statistical comparisons among data sets; the
second was used to evaluate the feasibility of applying indices to canopy
coverage data.
Further, the number-based indices were calculated with a modification of
the Shannon-Weaver function which since has been discontinued. Thus, direct
comparisons between indices are not legitimate, but relative tendencies
should be meaningful.
SPECIES DIVERSITY (CANOPY COVER) FOR THE COLSTRIP STUDY SITES
Diversity indices were calculated using coverage data from all sites and
dates of sampling.
Canopy cover diversity for the Colstrip locations are presented in
Figures 2.1, 2.2, and 2.3, showing data for 1974, 1975, and 1976, respec-
tively. There are definite diversity differences among sites, and they are
quite consistent over years. Magnitudes change, but relative diversity is
reasonably constant.
3.0
2.5 -
2.0 -
1.5
JULY
AUGUST
SEPTEMBER
HAY KLUVER KLUVER KLUVER KNOLL KNOLL KNOLL
COULEE EAST WEST NORTH ABC
Figure 2.1. Shannon-Weaver function (HP) for Colstrip Study sites, 1974.
43
-------
3.0
2.5 -
Q.
I
2.0
1.5
1 1 1 1 1
HAY KLUVER KLUVER KLUVER KNOLL
COULEE EAST WEST NORTH A
JUNE
JULY
AUGUST
1 1
KNOLL KNOLL
Figure 2.2. Shannon-Weaver function (HP) for Colstrip Study sites, 1975.
3.0 -i
2.5
2.0 -
1.5
1 1 • —I 1 1 1 1
HAY KLUVER KLUVER KLUVER KNOLL KNOLL KNOLL
COULEE EAST WEST NORTH ABC
Figure 2.3. Shannon-Weaver function (HP) for Colstrip Study site, August,
1976.
44
-------
These site differences can be related to plant species dominance and
density. For example, Kluver West has a low diversity due to the strong
dominance of needle-and-thread grass (Stipa comata) and the paucity of other
species. Soil and microclimate variation, as well as grazing history, are
the likely causes of this situation. Thus, comparisons among sites probably
are of 1imited value.
The McRae Knolls, which are adjacent sites of similar grazing history
and climate show diversity differences because of edaphic factors among
sites. This is reflected primarily in species composition. When analyzed
for range condition, Knolls A and C were in good condition and Knoll B was
excellent. This estimate was based on percent of climax vegetation for the
sites. The fact that the diversity indices show a similar pattern suggests
a relationship between range condition and diversity, which is consistent
with the results of many other studies.
Seasonal differences in diversity follow previously reported trends
(Taylor et al. , 1976). As the season progresses, many of the ephemeral
forbs and cool-season grasses disappear or diminish in their contributions
to total canopy coverage.
PHENOLOGY STUDIES
Phenology data has been recorded since 1974 to establish a baseline
data set.
The authors hypothesize that phenology will be a less sensitive approach
than diversity in detecting pollution effects. This is primarily because of
the large sample size required and the need to accumulate observations over
a number of years in order to discriminate between "normal" and "abnormal"
growth patterns. Therefore, the authors are continuing the data collection,
and in fact are increasing the frequency of sampling, especially in the
spring and early summer.
PROCEDURES
A phenologic scorecard designed by the authors was used (Taylor et al_. ,
1975); the categories are shown in Table 2.1. This classification has been
modified since last season, with the insertion of the early flowering stage.
With this addition, the system seems to fit a number of vegetation types and
has generated considerable interest from various fields of ecological re-
search. On each visit to an experimental site, a reconnaissance survey is
made. Each recognizable plant species is characterized as to its modal
phenologic stage. The scorecard approach attempts to quantify significant
growth stages without resorting to counts or measurements.
The data are stored for further comparison and future analyses.
45
-------
TABLE 2.1. PHENOLOGY CODES.
Code Stages
1 Cotyledon (newly germinated)
2 Seedling
3 Basal Rosette
4 Early greenup, veg. buds swelling
5 Vegetative growth, twig elongation
6 Boot stage, flower buds appearing
7 Shooting seed stalk, floral buds opening
8 Early flowering
9 Flowering, anthesis
10 Late flowering
11 Fruit formed
12 Seed shatter, dehiscence
13 Vegetative maturity, summer dormancy, leaf drop
14 Fall greenup
15 Winter dormancy
16 Dead
DISCUSSION
For purposes of elucidation, the index values (stages) for Kluver North
are displayed graphically in Figures 2.4 (grasses) and 2.5 (forbs). The
differences among species curves for grasses are clearly related to longevity
and typical growth patterns (cool- vs. warm-season species).
Forbs and shrubs show more interspecific variation in phenology than do
grasses, but site differences are inconsistent. This probably is due to the
unusually favorable growing conditions during summer, 1975, following the
later spring. The authors recognize that since the stages comprise an un-
sealed, arbitrary series, the resulting graphs should be viewed only as gen-
eralized profiles of plant growth and not subjected to quantitative analysis.
46
-------
DEAD
WINTER DORMANCY
FALL GREENUP
MATURITY
SEED SHATTER
FRUIT FORMED
LATE FLOWERING
FLOWERING
FLOWER BUDS OPENING
SHOOTING SEED STALK
FLOWER BUDS APPEARING
BOOT STAGE
VEGETATIVE GROWTH
EARLY GREENUP
BASAL ROSETTE
SEEDLING
COTYLEDON
Agropyron smithii
Aristida longiseta
Koe/eria cristata
Poo sandbergii
Stipa coma fa
ANNUAL BROMES
/NEW SEEDLINGS, \
\ANNUAL BROMES/
MAY
14
JUNE
12
JULY
13
AUG. SEPT.
30 10
NOV.
I
Figure 2.4. Phenological profile of selected species in Kluver North
exclosure, 1975.
47
-------
DEAD
WINTER DORMANCY
FALL GREENUP
MATURITY
SEED SHATTER
FRUIT FORMED
LATE FLOWERING
FLOWERING
FLOWER BUDS OPENING
SHOOTING SEED STALK
FLOWER BUDS APPEARING
BOOT STAGE
VEGETATIVE GROWTH
EARLY GREENUP
BASAL ROSETTE
SEEDLING
COTYLEDON
O
Artemisia frigida
Plantago pafagonica
• — Sphaeralcea coccinea
Taraxacum officinale /NFW GROWTH\
Tragopogon dubius
\ TRDU /
MAY
14
JUNE
12
JULY
13
AUG. SEPT.
30 10
NOV.
I
Figure 2.5. Phenological profile of selected species in Kluver North
exclosure, 1975.
48
-------
The development of analytical procedures whereby quantitative compari-
sons can be made among species, locations, seasons, and treatments is a
continuing part of this research.
PLANT COLLECTIONS
An ongoing program of plant collection has been under way since the
initiation of the field phases of the project. Voucher species are deposited
in the Herbarium of Montana State University. In addition, field specimens
are maintained for reference of crews and others for consistency of naming,
correctness of identification, elimination of duplicate numbers in Unknowns,
etc.
Small field specimens
dure of Burleson (1975).
are taken in plastic covers, following the proce-
REFERENCES
Burleson, W. H. 1975. A method of mounting plant specimens in the field.
J. Range Manage. 28:240-241.
Daubenmire, R. R. 1959. A canopy-coverage method of vegetational analysis.
Northw. Sci. 33:43-64.
Shannon, C. and W. Weaver. 1949. Mathematical
Univ. Illinois Press, Urbana. 117 p.
theory of communication.
Taylor, J. E., W. C. Leininger, and
studies near Colstrip, Proc.
Sci. pp. 537-551.
R. J. Fuchs. 1975. Baseline vegetational
Ft. Union Coal Field Symp., Mont. Acad.
Taylor, J. E., W. C. Leininger, and R.
community changes due to emissions
eastern Montana. Section II of the
fired power plant, Second Interim
Series EPA-600/3-76-013. pp. 14-40.
J. Fuchs. 1976. Monitoring plant
from fossil fuel power plants in
bioenvironmental impact of a coal-
Report. USEPA Ecological Research
49
-------
SECTION 3
SOIL AND EPIPHYTIC LICHEN COMMUNITIES
OF THE COLSTRIP, MONTANA AREA
by
S. Eversman
INTRODUCTION
Numerous field studies have used lichen distribution patterns to locate
areas of air pollution. The studies show that fewer lichen species occur in
polluted areas (LeBlanc and Rao, 1975), and that lichen species transplanted
into polluted areas lose vigor and/or die (LeBlanc and Rao, 1975). The
Colstrip project in southeast Montana provides the rare opportunity to map
lichen communities and obtain baseline physiological and anatomical
information before the two 350 megawatt coal-burning power plants begin
operation. This project will be able to establish the time progression of low-
level pollution effects on a previously pristine area.
The specific objectives are to:
1) Determine baseline lichen community information for the
grassland and ponderosa pine vegetation types.
2) Establish the baseline anatomical and physiological
condition of two native lichen species.
3) Continually monitor these characteristics to detect changes
caused by increasing air pollution.
4) Compare lichen monitoring results with measurements taken
simultaneously on associated vascular vegetation by other
investigators.
LOCATIONS AND METHODS
The lichen study sites in the Colstrip area are shown in Figure 3.1.
Table 3.1 provides this information for each site.
Samples of all observed lichen and moss species on soil and rocks have
been collected from sites G1-G7 (grassland sites 8-97 km from the Colstrip
power plants). Specimens were taken outside the exclosure fences at the common
CFPP project research sites at Hay Coulee, Kluver North, Kluver West, Kluver
East, (Gl + G3-G5 on Figure 3.1).
50
-------
Forsyth
Rosebud County
Treasure
County
Sarpy
Creek
PI
P2
P3 /^P7
9 G4/
P^ ^G3'
^sf \~G5
r
>P8
Big Horn County
Lame Deer
Northern Cheyenne
Reservation
P9
Ashland*
Custer County
Pll
P12|
P16
I—JP15
Birneyj
I
I
I
I
I
"h
L
>P13
—-^
»P14
Custer
National Forest
TC
G7
1 cm-8.0km
Figure 3.1. Location of Field Study Sites in the Colstrip area. P1-P16 are
ponderosa pine sites on ridges; G1-G7 are grassland sites.
51
-------
TABLE 3.1. DESCRIPTION OF LICHEN COLLECTION SITES, COLSTRIP AREA (Name; air
distance and direction from Col strip; exposure for ponderosa
pine sites; location).
Grassland Sites
Gl Hay Coulee; 11.6 km SE; TIN, R42E, Sec. 28
G2 McRae Knolls; 13.8 km SE
G3 Cow Creek (Kluver West); 11.6 ESE; TIN, R42E, Sec. 2
G4 North Pasture (Kluver North); 14.7 km E; TIN, R43E, Sec. 6
G5 School Section (Kluver East); 18.3 km ESE; TIN, R43E, Sec. 15
G6 Harvey Sage Site, BLM land along Cow Creek Road; 8 km SE
G7 Pasture adjacent Cow Creek Road-Otter Creek Road intersection; 70 km SSE
Ponderosa Pine Sites (Ridges)
PI Sarpy Creek, Charles May Ranch; 44 km W; ENE exp; T2N, R37E, Sec. 36
P2 Castle Rock; 16 km W; E, NE exp; TIN, R41E, Sec. 36
P3 Kluver NE1; 9.6 km NE; SW exp; T2N, R42E, Sec. 16
P4 Kluver El; 10 km E; W exp; T2N, R42E, Sec. 29
P5 D.McRae hill; 9.6 km SSE; NNW exp; TIN, R42E, Sec. 36
P6 Ridge direction south of Kluver West grassland exclosure (G3); 11.6 km S;
NW exp; TIN, R42E, Sec. 2
P7 Diamond Ranch Buttes; 28 km ENE; SW exp; T2N, R43E, Sec. 22
P8 Morning Star View picnic ground, North Cheyenne Indian Reservation; 26 km
SSE; N exp; T2S, R41E, Sec. 12
P9 Stop along Highway 212 between Lame Deer and Ashland; 35 km SE; NE exp.
P10 East Otter Creek Divide Road; 54 km SE; NW exp; T2S, R46E, Sec. 24
Pll SEAM Site 1; 52 km SE; W exp; T2S, R46E, Sec. 22
P12 SEAM Site 2; 50 km SE; NW exp; T2S, R46E, Sec. 22
P13 Home Creek Butte; 60 km SE; NW exp; T2S, R46E, Sec. 4
P14 Ridge near Three Mile Creek, Lemonade Spring Road; 90 km SE; NW exp; T2S,
R46E, Sec. 4
P15 Fort Howes Ranger Station; 65 km SSE; N exp; T6S, R45E, Sec. 19
P16 Poker Jim Butte; 64 km S; NW exp; T6S, R44E, Sec. 17
TC Taylor Creek Grassland Fumigation Site; 18 km ESE of Fort Howes Ranger
Station, Powder River Co.; T7S, R47E, Sec. 9
52
-------
Ten fenceposts, serving as permanent observation points, were placed
inside each of the exclosures where grass cover was the lowest and lichen cover
was the greatest. The point-drop method (Hanson, 1950) was used to record
ground cover (bare soil, grass, litter, moss, forb, lichen species) at each
fencepost. The center of the point-drop frame was placed on the north side of
each fencepost and 200 points per exclosure were recorded.
Sixteen ponderosa pine study sites (P1-P16) are located on ridges 8-97 km
from Colstrip; 13 of them are exposed ridges that face toward the power plant
stacks. Ten sites (P1-P5, P8, P11-P14, P16) have sulfation plates placed there
by other researchers (Gordon, 1975; Gordon et al_. , 1976). Site PI0 was used as
a control and source of transplant material; P9 was used as a transplant source
in 1975 only. Samples of lichens and mosses from all substrates have been
collected from all sites.
Ten trees on each site (excluding P9) are permanently marked for annual
determination of percentage of cover and frequency of each lichen species
present. Cover classes 1 - 5 (1 = 1 - 5% cover; 2 = 6- 25%; 3 = 26 - 50%; 4 =
51 75%; 5 = 76 - 100%) are recorded for each species encountered on each of
the four compass sides of each tree (north = northeast to northwest; east =
northeast to southeast; etc.). Midpoints of each cover class are used for
calculations (Tables 3.3 and 3.4).
Transplants to some sites were made by wiring ponderosa pine branches
covered with Usnea hirta onto selected branches of 10 pine trees facing toward
the power plant. Site P10 was the source of all transplant material in 1975 and
1976. Transplants were made to sites P1-P6 to augment local U. hirta
populations available for collection and to place sampling specimens in
locations more exposed than those of existing populations. Transplants were
made to sites P8, P15, and P16 to provide comparisons with the native
populations to detect effects of transplanting. Site P7 had no epiphytic
1ichens at al 1.
Parmelia chlorochroa or Usnea hirta samples are collected periodically
from test sites~(except P9) for manometric determination of respiration rates,
analysis of total sulfur (as S04) content by the Montana State University Soils
Testing Laboratory (Eversman, 1975), and for microscopic examination of algal
color and plasmolysis.
Plasmolysis percentage is determined by preparing three slides of three
different plants from each site, counting 100 algal cells on each slide, and
recording the number of plasmolyzed cells out of each 100 counted.
RESULTS
Table 3.2 lists the primary lichen species on seven grassland sites.
Cladonia spp. squamules are the dominant lichen types on the soil at all
exclosure sites; species identification has not been made because the
reproductive structures are very rarely found. Therefore, the other most
abundant species, Parmelia chlorochroa, is used for analytical work.
53
-------
on
TABLE 3.2. GROUND COVER OF THE PRIMARY LICHEN SPECIES ON 7 GRASSLAND SITES. PERCENTAGES DETERMINED BY
THE POINT-DROP METHOD. + = present, less than 0.5% x = present in undetermined quantities.
Lichen
Cladonia spp.
Parmelia chlorochroa
Collema tenax
Fulgensia fulgens
Buellia epigaea
Dermatocarpon lachneum
Squamarina lentigera
Lecidea decipiens
Acarospora schleicheri
Tonim'a coeruleonigricans
Agrestia hispida
Hay
Coulee
Gl
23.3%
16.7
5.0
+
+
+
+
+
+
+
+
McRae
Knolls
G2
X
X
X
X
X
X
Kluver
West
G3
27.0
16.5
2.5
0.5
+
+
Kluver
North
G4
24.0
9.5
2.0
1.0
+
+
Kluver
East
G5
13.6
10.0
2.7
+
+
+
+
Harvey
G6
X
X
X
X
X
X
X
X
X
Pasture
G7
X
X
X
X
-------
TABLE 3.3. PONDEROSA PINE SITES: PERCENTAGE OF TOTAL LICHEN COVER,
FREQUENCY (Number of Trees Having Lichens), NUMBER OF
EPIPHYTIC LICHEN SPECIES AT THAT SITE.
Site
P10, East Otter Creek
PI 2, SEAM Site 2
P14, 3-Mile
Pll, SEAM Site 1
P16, Poker Jim
PI 3, Home Creek Butte
P15, Fort Howes
P5, D. McRae
P3, Kluver NE1
P2, Castle Rock
PI , Sarpy Creek
P4, Kluver El
P7, Diamond Buttes
% cover
39.75
21.00
15.25
13.63
12.88
11.75
10.81
9.13
6.88
6.38
1.75
1.13
0
Frequency
10
10
10
10
10
10
10
8
8
8
4
2
0
Number of
species
1
5
4
6
6
7
7
8
7
9
3
4
0
55
-------
TABLE 3.4. LICHEN COVER OF PONDEROSA PINE TREE TRUNKS, COLSTRIP AREA. (Percentage cover/frequency, 10
trees sampled on each site. Site P7 has 0 for all species.)
en
CTl
Lichen
Usnea
hi rta
Parmel ia
sulcata
Parmel i ops is
ambigua
Alectoria
f uscescens
Hypogymnia
physodes
Parmel ia
infumata
N
S
E
W
N
S
E
W
N
S
E
W
N
S
E
W
N
S
E
W
N
S
E
W
P10
12.0/10
11.1/8
7.4/9
9.3/10
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
P12
5.9/10
2.8/7
6.6/9
1.8/7
1.9/5
0.1/1
0.4/3
0
0
0
0
0
0.3/2
0
0.1/1
0
0.9/2
0
0.1/1
0
0
0
0
0
P14
5.9/10
0.8/6
3.3/7
0.8/6
1.8/4
0
0
0.3/2
0.1/1
0
0
0
2.1/7
0
0.1/1
0.1/1
0
0
0
0
0
0
0
0
Pll
3.1/10
3.1/7
4.6/9
0.9/5
0.3/2
0
0.3/2
0
0.3/2
0
0.1/1
0.1/1
0.1/1
0.3/1
0.3/2
0
0
0
0
0
0
0
0
0
P16
3.9/10
0.3/2
1.1/6
1.1/7
2.3/4
0.8/1
0
0
0.8/1
0
0
0
0.4/3
0
0
0.4/3
0
0
0
0.1/1
0.1/1
0
0.8/1
0
P13
2.1/10
0.8/3
3.0/9
0.3/2
0.3/2
0
0.3/2
0
1.0/3
0
1.0/1
0
0.3/2
0
0.1/1
0
0.9/3
0
0
0.1/1
0
0
0
0
P15
3.1/10
0
0.1/1
0.5/4
0.3/2
0
0
0.3/2
3.5/8
0
0.4/3
0.4/3
0.6/6
0
0
0
0.3/2
0
0
0.1/1
0.3/2
0
0
0.1/1
P5
0.9/7
0
0.3/1
0
0.3/2
0
0
0
3.5/8
0
1.6/3
0.1/1
0.6/5
0
0.3/2
0
0.9/2
0
0.1/1
0
0.4/3
0
0.1/1
0
P3
1.6/9
0.3/1
0
0.4/2
0.4/3
0
0
0
3.6/5
0
0.3/1
0.1/1
0.5/4
0
0
0
1.4/6
0
0
0.1/1
0.3/1
0
0.1/1
0
P2
1.0/8
0.1/1
0.1/1
0.3/2
0.3/2
0
0
0
1.4/5
0
0
0
0.1/1
0
0
0
0.5/4
0
0
0.1/1
1.0/4
0
0.1/1
0
PI
0.4/3
0
0
0
0
0
0
0
0.9/2
0
0.1/1
0.1/1
0.1/1
0
0
0
0
0
0
0
0
0
0
0
P4
0.8/1
0
0.1/1
0
0
0
0
0
0.1/1
0
0
0
0
0
0
0
0.1/1
0
0
0
0
0
0
0
(continued)
-------
TABLE 3.4. (continued)
Lichen
Lecanora
subfusca
Cetraria
pinastri
Parmel ia
ulophyl lodes
Letharia
vulpina
Total epiphyte
P10
N
S
E
W
N
S
E
W
N
S
E
W
N
S
E
W
cover:
39
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
.75
P12
0
0
0
0
0
0
0
0
0.3/2
0
0
0
0
0
0
0
21.00
P14
0
0
0
0
0
0
0
0
0
0.1/1
0
0
0
0
0
0
15.25
Pll
0
0
0.1/1
0
0
0
0.1/1
0
0
0.1/1
0
0
0
0
0
0
13.63
P16
0
0
0
0
0
0
0
0
0.9/2
0.1/1
0
0
0
0
0
0
12.88
P13
0
0
0
0
0.4/1
0
0
0
1.1/3
0
0
0
0
0
0
0
11.75
P15
0.1/1
0
0.4/3
0.4/3
0
0
0
0
0.1/1
0
0
0
0
0
0
0
10.81
P5
0.9/1
0
0
0
0.4/1
0
0
0
0
0
0
0
0
0
0
0
9.13
P3
0
0
0
0
0
0
0
0
0
0
0.1/1
0
0.1/1
0
0.1/1
0
6.88
P2
2.4/4
0
0
0
0.1/1
0
0
0
0
0
0
0
0.1/1
0
0
0
6.38
PI
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1.75
P4
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1.35%
-------
Tables 3.3 and 3.4 give the epiphytic lichen community data collected and
compiled through 1976. Only ten species are present consistently enough to
appear in the sampling. The species list includes names of other collected
epiphytic lichens.
Respiration rates for Usnea hirta samples for 1975 and 1976 are in Table
3.5; those for Parmelia chlorochroa are in Table 3.6. Results from sulfate
sulfur analyses for 1974 and 1975 are in Table 3.7; analyses for 1976 samples
have not yet been completed.
Determination of some baseline algal plasmolysis counts have been
completed from the P10 site. The plasmolysis values (means ± standard
deviation) are: 1975, September, 17 ±2; 1976, March, 3 ±3; July, 11 ±2; July, 9
±3. Counts are continuing for all other collection.
DISCUSSION
After the summer of 1975, it appeared that recording only the
presence/absence data for epiphytic lichen communities was not sensitive
enough to detect and monitor the low-level chronic S02 stress, despite reports
using these criteria from other regions (LeBlanc and Rao, 1975). However, in
this area any pollution stress will be newly-introduced and epiphytic growth is
slight compared to that in the more humid climates of other studies.
Therefore, the more detailed data were acquired during 1976. The marked trees
allow for annual repetition of measurements on the sites. Three more sites,
closer to the power plant, are being added for analysis beginning in 1977.
Determination of respiration rates is a method sensitive enough to detect
adverse pollution effects in U. hirta (Eversman, 1976). However, bacteria
populations increased and/or changed in S02-stressed Parmelia chlorochroa
plant bodies. This probably caused the respiration rate of the lichen to be
overestimated. Photosynthetic rate would be a more specific indicator of the
physiological condition of the lichen plant. This information combined with
the plasmolysis counts gives adequate information on the vitality of these
lichens. Eventually, the effects observed at the organismic level will be
reflected at the community level. Photographs of many communities are also
being taken regularly.
Ponderosa pine trees in the moister habitats, with shrubs Symphoricarpos
albus, Rhus trilobata, and Ribes spp., generally have the heaviest lichen cover
on the trunks and branches (P10, Pll, P12). Where the understory is dominated
by such species as Yucca glauca and Agropyron spicatum (sites PI, P3, P4, P6,
P7), the lichen cover is sparse and restricted to the lower 30 cm of trunks on
the north and east sides, if it appears at all. None of the sites in the Custer
National Forest required transplants for monitoring purposes.
Mosses, the only bryophytes encountered, have been collected from the
grasslands and from the soil and litter under the Ponderosa pines. Verified
identifications of these species, and of mosses and lichens collected from the
rocks, have not yet been completed.
58
-------
TABLE 3.5. RESPIRATION RATES OF Usnea Hirta SAMPLES, COLSTRIP AREA, 1975-76.
(Respiration rate is given as micro! iters of
weight/hour, manometrically determined.)
PI,
P2,
P3,
P4,
P5,
P6,
P7,
P8,
P9,
P10
Pll
P12
P13
P14
P15
P16
Site
Sarpy Creek
Castle Rock
Kluver NE1
Kluver El
D McRae
Kluver West transplants
(from P9, 1974)
Diamond Buttes
Morning Star View, transplants
natives
transplants
Road
, East Otter Creek
, SEAM 1
, SEAM 2
, Home Creek Butte
, 3-Mile, top of ridge
bottom of ridge
top of ridge
bottom of ridge
, Fort Howes (transpl from P9)
natives
ii
M
transplants
Poker Jim Butte transplants
' ii
natives
ii
n
Col lection
Date
9-25-75
9-14-76
9-25-75
9-14-76
9-25-75
9-15-76
9-14-75
9-15-76
9-15-75
9-15-76
7-16-76
6-24-76
9-25-75
n
11-20-76
5-01-76
8-11-75
9-25-75
3-23-76
6-23-76
7-15-76
8-09-76
9-15-76
7-16-75
7-15-76
7-16-17
7-15-76
7-16-75
7-15-76
7-16-75
n
8-09-76
II
5-01-75
II
5-13-76
9-15-76
n
9-25-75
9-15-76
7-17-75
7-16-76
9-15-76
X
640*
658
640*
499
640*
633
640*
573
640*
827
669
744*
640*
828
661
563
508
640
709
744
463
668
633
545
453
548
498
462
486
539
528
667
547
829
510
576
470
597
640*
669
630
610
654
02 consumed/g dry
1 s.d.
40
129
40
86
40
98
40
53
40
38
50
45
40
135
60
109
51
40
63
45
57
41
53
35
19
27
48
55
15
64
38
76
40
58
213
113
50
43
40
74
77
31
52
* respiration rate of duplicate material when transplanted from P10 site.
59
-------
TABLE 3.6. RESPIRATION RATES OF Parmelia chlorochroa SAMPLES, COLSTRIP AREA,
1974-6. (Respiration rate is in micro! iters
wt/hour, manometrically determined.)
02 consumed/g dry
Col lection
Site Date
Gl , Hay Coulee 7-20-74
8-30-74
6-26-75
5-14-76
7-16-76
9-14-76
G2, McRae Knolls 9-15-76
G3, Kluver West 7-21-74
7-07-75
7-07-75
7-14-76
9-14-76
G4, Kluver North 7-21-74
7-07-75
7-16-75
9-14-76
G5, Kluver East 7-21-74
6-26-75
7-14-76
9-15-76
9-15-76
G6, Harvey 1 9-15-74
9-15-74
G7, Pasture 5-01-75
5-14-76
X
314
332
215
458
290
274
341
218
349
293
272
301
342
349
242
262
227
316
322
404
239
339
321
337
371
1 s.d.
14
43
26
43
19
22
21
16
21
29
25
15
18
21
4
34
34
40
26
32
15
46
28
132
54
60
-------
TABLE 3.7. SULFUR (AS TOTAL SULFATE) CONTENTS OF Usnea hirta AND Parmelia
chlorochroa SAMPLES COLLECTED IN 1975. (Determined by MSU
Soils Testing Laboratory.)
Site
Collection
Date
Usnea hirta
% sulfur
as sulfate
1 s.d.
P9, Road 5-01-75
P15, Fort Howes transplants 5-01-75
P15, Fort Howes natives "
P9, Road 8-11-75
P3, Kluver NE1 9-25-75
P8, Morning Star View "
P13, Home Creek Butte 7-16-75
P10, East Otter Creek 7-16-75
P10, East Otter Creek 9-25-75
Parmelia chlorochroa (includes 1974)
Gl, Hay Coulee
G3, Kluver West
G4, Kluver North
G5, Kluver East
G6, Harvey
Taylor Creek
G7, Pasture
7-21-74
9-15-74
5-01-75
0.25
0.25
0.19
0.26
0.03
0.19
0.15
0.08
0.22
0.26
0.22
0.20
0.26
0.23
0.17
0.19
0.04
0.03
0.04
0.04
0.01
0.08
0.16
0.01
o.n
0.02
0.02
0.02
0.02
0.01
0.02
0.03
ACKNOWLEDGEMENTS
This project is funded by EPA grant No. R803213. Appreciation is
expressed to Drs. W.A. Weber and J.W. Thomson for assistance with lichen
identification; to Drs. C.C. Gordon, Jerry Bromenshenk, and Clint Carlson for
field assistance and to the landowners for permitting access to the research
sites.
61
-------
REFERENCES
Bird, C.D. Keys to the Lichens of West-Central Canada. The Herbarium,
Department of Biology, University of Calgary, 1970.
Eversman, Sharon. Lichens as Predictors and Indicators of Air Pollution from
Coal-Fired Power Plant Emissions. The Bioenvironmental Impact of a Coal-
Fired Power Plant, Second Interim Report, Colstrip, Montana. Corvallis
Environmental Research Laboratory, Office of Research and Development,
U.S. Environmental Protection Agency, Corvallis, Oregon, 1975.
Eversman, Sharon. Effects of Low-Level S02 Stress on Two Lichen Species. The
Bioenvironmental Impact of a Coal-Fired Power Plant, Third Interim
Report, Colstrip, Montana. Corvallis Environmental Research Laboratory,
Office of Research and Development, U.S. Environmental Protection Agency,
Corvallis, Oregon, 1975.
Gordon, C.C. Investigations of the Impact of Coal-Fired Power Plant Emissions
upon Plant Disease and upon Plant-fungus and Plant-insect Systems. The
Bioenvironmental Impact of a Coal-Fired Power Plant, Second Interim
Report, Colstrip, Montana. Corvallis Environmental Research Laboratory,
Office of Research and Development. U.S. Environmental Protection
Agency, Corvallis, Oregon, 1975.
Gordon, C.C., Clinton Carlson, Phillip Tourangeau. A Cooperative Evaluation of
Potential Air Pollution Injury and Damage to Coniferous Habitats on
National Forest Lands near Colstrip, Montana. Interim Report of
Activities from June 1, 1975 to May 30, 1976. USDA, Forest Service,
Northern Region and University of Montana, Environmental Studies
Laboratory. Missoula, Montana, July, 1976.
Hale, Mason. How to Know the Lichens. Wm.C. Brown, Dubuque, Iowa
1969.
Hanson, Herbert C. Ecology of the Grassland. II. Botanical Review 16: 283-
360, 1950.
LeBlanc, Fabius and D. Rao. Effects of Air Pollutants on Lichens and
Byrophytes. In: Mudd, J.B. andT.T. Kozlowski (eds). Responses of Plants
to Air Pollution. Academic Press, New York, 1975.
Wetmore, Clifford. Lichens of the Black Hills of South Dakota and Wyoming.
Publications of the Museum, Michigan State University, East Lansing,
Michigan, 1967.
62
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APPENDIX
LIST OF LICHEN SPECIES COLLECTED FROM THE COLSTRIP, MONTANA, AREA;
BIG HORN, ROSEBUD, AND POWDER RIVER COUNTIES.
(Collection number(s); site; substrate.)
1. Acarospora schleicheri (Ach. ) Mass. 565, Gl. Soil.
2. Agrestia hispida (Mereschk.) Hale & W.Culb. 569, G6; 609, Sarpy Creek-
720, P7; G5. Soil.
3. Alectoria fuscescens 684, Upper Indian Creek Divide; 690, P3; 694, P14;
556, Ash Creek. Ponderosa pine base. det. I.M. Brodo.
4. Caloplaca aurantiaca (Lightf.) Th.Fr. 547, P6. Decorticated stump.
5. Candelariella aurella (Hoffm.) Zahlbr. 710, P16; 744, Pll. Rock.
6. Cetraria pinastri (Scop.) S. Gray. 548, Ash Creek; 733, Pll. Pine tree
trunk.
7. Cladonia pyxidata (L. ) Hoffm. 543, 544 557, G3; 571, G6. Soil.
8. Cladonia spp. squamules All sites. Soil.
9. Collema tenax (Sw.) Ach. 580, Ash Creek; 581, G4; 582, Gl. Soil.
10. Cyphelium notarisii 549, P6. Decorticated stump.
11. Dermatocarpon lachneum (Ach.) A.L.Sm. 584, Gl. Soil.
12. Everm'a mesomorpha Nyl. 598, P15; 660, P10. Ponderosa pine base.
13. Fulgensia bracteata (Hoffm.) Ras. with F. fulgens (Sw.) Elenk. 716, P7;
546, 545, G3; 560, Gl; 568, G6. Soil.
14. Hypogymnia physodes (L.) W. Wats. 538, P15; 539, Ash Creek; 688, P3; 698,
-P16; 703, P8. Pine tree trunk.
15. H. bitten' (Lynge) Ahti. 676, P16. Ponderosa pine trunk.
16. Lecanora calcarea (L.) Somm. 608, P16. Rock.
17. L. chrysoleuca (Sm.) Ach. 669, P10. Rock.
18. L. melanophthalma (Ram.) Ram. 709, P16; 669, P10. Rock.
19. L. mural is (Schreb.) Rabenh. 665, P8. Rock.
20. Lecidea decipiens (Hedw.) Ach. 566, G6; 567, Gl. Soil.
21. L. marginata Schaer. 682, Upper Indian Creek; 705, P16; 732, P14. Rock.
22. L. rubiformis (Wahlenb. ex. Ach.) Wahlenb. 610, Sarpy Creek; 648, P16;
704, P5; 715, P7; 731, P16. Soil in rock crevices.
23. Letharia vulpina (L. ) Hue. 654, P6; 597, P15; 689, P3. Pine tree trunks
24. Omphalodiscus virginis (Schaer.) Schol. 693, P8. Rock.
25. Parmelia chlorochroa Tuck. G3; 559, Gl ; 653, PI; 656, G5; 658, Sarpy
Creek. Soil.
26. P. infumata Nyl. 554, Ash Creek; 674, P16; 738, 745, Pll; 729, P16. Pine
tree trunk.
27. P. elegantula (Zahlbr.) Szat. 729, P16. Pine tree trunk.
28. P. lineola Berry 666, 671, P10. Rock.
29. P. subdecipiens Vain. 562, P15; 652, PI. Rock.
30. P. subolivacea Nyl. 553, 557, Ash Creek. Pine tree trunk, branches.
31. p. sulcata Tayl. 552, Ash Creek; 675, P16; 696, P14; 735, Pll. Pine tree
trunk.
63
-------
32. P. ulophyl lodes (Vain.) Sav. 563, Ash Creek; 564, P15; 695, P14; 699,
P16. Pine trunk.
33. Parmeliopsis ambigua (Wulf.) Nyl. 605, P3; 672, P10; 685, Upper Indian
Creek; 691, P3; 734, Pll. Base of pine trees.
34. Physcia caesia (Hoffm.) Hampe 558, P15. Rock.
35. Peltigera canina (L.) Willd. 579, P15. Soil.
36. P. rufescens (Weis.) Humb. 692, P3. Soil.
37. Rinodina confragosa (Ach.) Koerb. 702, PI. Rock.
38. Squamarina lentigera (G.Web.) Poelt. 570, Gl; 718, P7. Soil.
39. Toninia caeruleonigricans (Lightf.) Th.Fr. 714, P7. Soil.
40. Usnea hirta (L.) Wigg. 603, Whitetail Guard Station; 550, Ash Creek; 572,
P15; 686 Upper Indian Creek.
41. Umbilicaria torrefacta (Lightf.) Schrad. 740, Pll. Rock.
42. Umbilicaria vellea (L.) Ach. 608, Sarpy Creek. Rock.
43. Verrucaria sp. 706, P16. Rock.
44. Xanthoria elegans (Link.) Th.Fr. 609, P10; 707, P16. Rock.
45. X. polycarpa (Ehrh.) Oliv. 673, P15. Shrub branches.
64
-------
SECTION 4
INVESTIGATION OF THE IMPACT OF COAL-FIRED POWER PLANT EMISSIONS UPON THE
DISEASE/HEALTH/GROWTH CHARACTERISTICS OF PONDEROSA PINE-SKUNKBUSH ECOSYSTEMS
AND GRASSLAND ECOSYSTEMS IN SOUTHEASTERN MONTANA
by
C. C. Gordon, P. C. Tourangeau, and P. M. Rice
INTRODUCTION
This portion of the EPA-CERL (Corvallis Environmental Research Labora-
tory, Oregon)-sponsored studies was initiated to ascertain and quantify the
impact of coal-fired power plant emissions upon the two major ecosystems of the
Col strip area of southeastern Montana: (1) Ponderosa pine-skunkbush (Pinus
ponderosa-Rhus trilobata) and (2) cool season-short grass. Before the two, 350
MW capacity coal-fired power plants were constructed and began operations at
Col strip, air monitoring studies by federal and state agencies, the utility
companies, and private research institutes established that ambient air in the
Col strip area was as pristine and free of phytotoxic gases as any area in the
contiguous United States thus far studied (Northern Cheyenne Research Project,
1976). Because of the pristine nature of this area of Montana, it was
hypothesized that if the baseline characteristics of the chemical, physical,
and growth/health/disease parameters of indigenous flora and fauna species
could be established and quantified before the power plants began operations,
many future direct and indirect impacts of the atmospheric emissions upon these
species could be measured and quantified. To test this hypothesis, several
study objectives were carried out. These encompassed the establishment of:
(1) Baseline levels of sulfur and fluoride concentrations in the domi-
nant indigenous plant species of both the ponderosa pine-skunkbush
and cool season-short grass ecosystems;
(2) baseline growth/health/disease parameters of ponderosa pine foliage;
(3) baseline insect population and insect damage parameters for ponde-
rosa pine of the area;
(4) baseline fungal population and damage to selected indigenous plant
species; and
(5) baseline physical (pH) and chemical parameters of precipitation.
These baseline parameters and characteristics were used to study species
of flora and fauna collected from permanently established ponderosa pine-
65
-------
skunkbush sites located at varying distances and directions from Colstrip
during the 1974 and 1975 growing seasons.
After the 1974 and 1975 baseline studies on the ponderosa pine-skunkbush
sites were completed (see EPA-CERL 1976 Second Interim Report, EPA-600/3-76-
013). some of the earlier study objectives were deleted (i.e., fungal popula-
tions and host-parasite relationships on indigenous plant species) and others
were modified (i.e. , the number of ponderosa pine-skunkbush sites and precipi-
tation chemistry studies). The modification or deletion of those original
objectives were initiated to allow concentration on those objectives which more
adequately tested the original hypothesis.
During 1975 and 1976, air monitoring by the Montana Department of Health
and Environmental Sciences (DHES) Air Quality Bureau at their monitoring site 4
km southeast of Colstrip disclosed that S02, N02, 03, and F concentrations in
the ambient air were all substantially below both federal and Montana state
standards for these pollutants (Maughan, 1977). Wind speed and direction data
collected by personnel of the DHES Air Quality Bureau at their Colstrip site
disclose that the prevailing winds during the growing season (May to September)
are to the southeast and east and to the west and northwest, and average
between two to six miles per hour.
Since September 1975, when Colstrip Unit 1 went on line, and July 1976,
when Unit 2 started up, these two units have operated at an average megawatt
capacity of 1/3 (33%) to 2/5 (40%) of their total rated capacity of 700+ MW.
The estimated atmospheric emissions of the Colstrip units when operating at 1/3
and 2/5 capacity are presented in Table 4.1 below.
TABLE 4.1. ESTIMATED ATMOSPHERIC EMISSIONS OF COLSTRIP UNITS.
Unit 1 & 2
at 1/3 capacity
Unit 1 & 2
at 2/5 capacity
Unit 1 & 2
at full capacity
S02 (tons/yr)
NO (tons/yr)
Particulate (tons/yr)
Fluoride (Ibs/yr)
6,046
6,920
536
4,666
7,256
8,304
644
5,600
18,142
20,760
1,612
14,000
Source: Montana Department of Health and Environmental Sciences, 1975
MATERIALS AND METHODS
COLLECTION OF VEGETATION
Five ponderosa pine-skunkbush sites located east to south of Colstrip at
distances of 5 to 80 km were utilized during both the 1975 and 1976 study
periods. The locations of each of these five sites are listed in Table 4.2.
66
-------
TABLE 4.2. LOCATIONS OF FIVE PONDEROSA PINE-SKUNKBUSH SITES.
Ponderosa Pine-
Skunkbush Sites
Plot
Plot
Plot
Plot
Plot
S-3
S-5
SE-2
SE-4
E-l
T2S,
T6S,
T1S,
T47E
T2N,
Location
R41E,
R44E,
R42E,
, R47E
R42E,
Sec
Sec
Sec
, Sec
Sec
Distance
from
NE 12
SE 8
SW 1
NW 19
SW 29
26
66
16
56
5
and Direction
Col strip
km
km
km
km
km
SSE
SSE
SSE
ESE
E
Ponderosa pine foliage was collected during the 1975 field season from these
plots for fluoride and sulfur analysis, evaluation of seven selected foliar
pathologies, determination of percent needle retention, and measurement of
needle length and fascicular cross-sectional area.
Each of the five plots consisted of ten permanently marked trees. Based
on tree height and diameter at breast height (DBH), five of the trees were
classified as "older" and five were classified as "younger." From each of
these ten trees, four branches were removed from the upper third of the crown
and four branches from the lower third of the crown from the side of the trees
facing the Colstrip steam generating complex. The branches from each tree were
placed in plastic sacks with appropriate identification of tree number and
crown position and transported to the field laboratory or to the Environmental
Studies Laboratory at the University of Montana for subsequent preparation for
chemical analyses, pathological evaluations, and measurements.
The percent needle retention for 1972, 1973, and 1974 year's foliage was
determined for each internode from each branch for the respective categories of
crown position and tree age by counting all fascicular scars on each internode,
then removing all retained fascicles, counting them, and computing the percent
needle retention as shown:
fascicles present -, x -|00
^-fascicles present + number of fascicular scars;
The mean percent needle retention for each year's internode was computed, and
subsequent data analyses used these basic values.
One hundred fascicles were randomly selected from the group of retained
fascicles from each internode, the fascicular sheaths were removed, and one
needle was randomly selected from each fascicle for pathological evaluation.
Ten fascicles were also selected for measurement of fascicular cross-sectional
area. After evaluation, 25 needles were randomly selected for measurement of
needle length.
67
-------
Each needle was evaluated for the presence of seven pathologies: (1)
percent basal necrosis, (2) percent basal scale, (3) percent defoliator, (4)
percent tip burn, (5) percent total necrosis, (6) percent mottled needles, and
(7) percent healthy needles. Percent healthy needles is the percentage of
total needles without any visible pathology whatsoever and, as such, are
completely green. Percent total necrosis is an estimate of the total necrotic
surface area of the 100 needles. This category includes chlorosis or yellow-
ing. The percent occurrence of each pathology was computed, and all data
analyses used these derived values.
The method of sampling results in the partitioning of nine categories of
tree age and crown position within each plot for each year's foliage is as
follows:
Upper Crown - Younger Trees
Upper Crown - Older Trees
Upper Crown - All Tree Ages
Lower Crown - Younger Trees
Lower Crown - Older Trees
Lower Crown - All Tree Ages
All Crown Positions - Younger Trees
All Crown Positions - Older Trees
All Crown Positions - All Tree Ages
These categories permit a detailed analysis of within-plot and between-plot
variability as a function of duration of foliage exposure. Studies have shown
that sulfur and fluoride are partitioned in greater concentrations in the
taller dominant vegetation at a site which is impacted by the emissions of a
coal-fired power plant and which includes vegetation species similar to those
discussed here (Gordon ert al. , 1977); other studies have demonstrated that
fluoride is partitioned in the upper crown of conifers. Any investigation of
the effects of coal-fired power plant emissions upon ponderosa pine sites in
southeastern Montana requires a background preoperational analysis of the
partitioning of fluoride and sulfur, pathologies associated with air pollu-
tion, needle length, and fascicular cross-sectional area in the upper and lower
crowns of ponderosa pine.
Understory species of grass, shrubs, and forbs were collected at the five
ponderosa pine-skunkbush sites during both the 1975 and 1976 summer collection
periods. A list of the understory species collected is presented in Table 4.3.
At each site, at least eight to ten different species of understory were
collected where possible. Collection of leaf foliage from each understory
species was obtained from a minimum of three plants and these were lumped and
tagged with the collection number, placed in plastic sacks, and brought back to
the laboratory for chemical analysis.
Needles manifesting the various needle pathologies (i.e., mottling,
weevil, fungal damage) were selected for histological studies. Histological
preparation and staining procedures have been described by Jones £t al_. (1965).
Photomicrographs were taken with the use of a Reichert Zetopan phase~microscope
adapted with Leitz cameras.
68
-------
TABLE 4.3. UNDERSTORY SPECIES.
Scientific
Common
Artemisia cana
Gutierrezia sarothrae
Prunus virginiana
Rhus trilobata
Festuca idahoensis
Agropyron spicatum
Artemisia 1udoviciana
Balsamorhiza sagittate
Andropogon scoparius
Lupinus sp.
Psoralea sp.
Yucca glauca
Aristida longiseta
Stipa comata
Vicia sp.
Artemisia frigida
Chrysothamnus nauseosus
Juniperus scopulorum
Artemisia tridentata
Silver Sage
Broom Snakeweed
Chokecherry
Skunkbush
Idaho Fescue
Bluebunch Wheatgrass
Prairie Sage
Arrow!eaf Balsamroot
Little Bluestem
Lupine
Scurf-pea
Yucca
Red Three Awn
Needle-and-thread
Vetch
Fringed Sage
Common Rabbit-brush
Rocky Mountain Juniper
Big Sage
CHEMICAL ANALYSIS OF VEGETATION
The methods of chemical analyses for sulfur and fluoride have been
reported in detail elsewhere (Kay et a_L , 1975; EPA, 1976). Therefore, these
procedures are summarized here.
Vegetation was dried under forced draft at 90°F, ground in a Wiley mill to
pass 30 mesh, and stored in capped plastic vials. For fluoride analysis, 0.50
g of dried ground plant material was slurried with CaO, charred under infrared,
and ashed overnight at 600°C in a muffle furnace. The ash was dissolved in 30%
HC10 made to 100 ml with 50% TISAB, and the fluoride activity was determined
within Orion specific ion electrode. The calculations for determining the
fluoride concentration in the original plant sample were computerized.
Sulfur determinations were carried out using a Leco induction furnace.
Before the data analysis, each derived variate (percents basal necrosis, basal
69
-------
scale, needle retention, tip burn, mottled needles, total necrosis, and healthy
needles) was coded by the arcsine transformation for percentages where:
coded variate = arcsine /variate x 57.29578
100
All coded variates and sample statistics were decoded for the presentation of
results.
RESULTS
DATA ANALYSIS OF VEGETATION
The data for both 1975 and 1976 were interrogated by Analysis of Variance
(ANOVA). The analyses were performed by the ANOVA computer program of Sokal
and Rohlf (1969).*
A four-level nested ANOVA was used because the 1975 data required tests
for each variable for: (1) Differences between plots; (2) differences between
upper and lower crown positions; (3) differences between the younger and older
trees, and (4) differences between the years of .foliage (1974, 1973, 1972).
The design for the four-level ANOVA is schematically shown in Table 4.4 for any
individual plot. In the actual analysis of all variables, the data for each
plot were arrayed exactly as in Table 4.4 and all plots were tested
simultaneously. In Table 4.4 the highest level, Level 4, is the individual
plot. At Level 3 are the two crown positions (upper and lower) within each
plot. At Level 2 are the tree ages (younger and older), and at Level 1 are the
three years of foliage (1974, 1973, 1972). The lowest level, Level 0, is the
individual variates, denoted as 1, 2, 3, 4, 5 to indicate the five samples
measured for fluoride content. Since each plot consists of five older and five
younger trees, the most fundamental data block in any plot would be the five
variates for a particular year's foliage for younger or older trees, from
either crown position. As can be seen from this design, tests are performed
between the years of foliage, between the tree ages, between the crown posi-
tions, and between the plots.
*The Program, as published in this edition, contains errors. Interested
readers should contact the authors of the text for current listings.
70
-------
TABLE 4.4. THE STRUCTURE OF THE FOUR-LEVEL NESTED ANOVA FOR EACH INDIVIDUAL
PLOT, 1975 COLLECTION, BOTH CROWN POSITIONS.
Level 4 - Plots
Level 3 - Crown Positions
Level 2 - Tree Ages
Level 1 - Age of Foliage
Level 0 - Five Individual
Observations for Each
Variable within Each
Age of Foliage
Plot
Upper
Lower
Younger
J_
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
Older
1
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
Younger
I
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
Older
1
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
71
-------
TABLE 4.5. FOUR-LEVEL ANALYSIS OF VARIANCE TABLE FOR FLUORIDE, 1975
COLLECTION.
Source
of
Variation
Sums
of
Squares
Degrees
of
Freedom
Mean
Squares
F
Ratio
Between the Plots 29.004
(Level 4)
Between Crown Positions
within the Plots 2.412
(Level 3)
Between the Tree Ages
within the Crown 4.876
Positions (Level 2)
10
7.251
.482
.487
15.031*
.989
1.032
Between the Years of
Fol iage within the
Tree Ages (Level 1)
Within the Years of
Foliage (Level 0)
18.889 40
103.868 240
.472 1.091
.432
*F ratio significant @ p ^ .05
TABLE 4.6. THE STRUCTURE OF THE THREE-LEVEL NESTED ANOVA FOR EACH INDIVIDUAL
PLOT, 1976 COLLECTION*
Level 3 Plots
Level 2 - Tree Ages within the Plot
Level 1 - Age of Foliage within the Tree Ages
Level 0 - Five Individual Observations for
Each Variable within the Ages of Foliage
Plots
Younger
!_
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
Older
1
1
2
3
4
5
2
1
2
3
4
5
3
1
2
3
4
5
*This design is precisely the same as that used for individual upper or lower
crown positions in the 1975 collection.
72
-------
The results of the four-level ANOVA for fluoride for the 1975 collections
are shown in Table 4.5, wherein the levels as detailed in Table 4.4 are listed
under Source of Variation.The only level in Table 4.5 which showed a signifi-
cant effect was Level 4, Between the Plots. The conclusion to be drawn from
Table 4.5 is that the means for the five plots are sufficiently different, that
they represent different populations as regards fluoride concentrations.
Also, it may be concluded that within the plots mean values for the respective
crown positions, tree ages, and individual years of foliage would represent the
same population.
In 1976, collections were obtained from upper crown positions only—
therefore the level supporting upper and lower crown for the ANOVA for 1975 is
not needed. The design for the 1976 collection is shown in Table 4.6 as a
three-level nested ANOVA*, where Level 3 is the individual plot. At Level 2
are the tree ages within the plots, at Level 1 are the individual years of
foliage within the ages, and at Level 0, the individual variates.
The results of ANOVA for fluoride for 1976 are shown in Table 4.7. None of
the F ratios are significant, and it may be concluded that fluoride means from
plots, tree ages, and years of foliage from the upper crown represent samples
from the same population.
TABLE 4.7. ANALYSIS OF VARIANCE TABLE FOR FLUORIDE, 1976 COLLECTION.
Source
of
Variation
Between the Plots
(Level 3)
Sums
of
Squares
3.912
Degrees
of
Freedom
4
Mean
Squares
.978
F
Ratio
2.025
Between the Tree Ages
within the Plots 2.415
(Level 2)
Between the Years of
Foliage within the 6.303
Tree Ages (Level 1)
Within the Years of 46.176
Foliage (Level 0)
20
120
.483
.315
.384
1.532
.819
The 1975 collections from the upper crown and the 1976 collections were
compared by a four-level nested ANOVA shown as Table 4.8, where, for brevity.
only two plots are shown. At Level 4 are the two collections, at Level 3 are
the plots within the collections, at Level 2 are the two tree ages within
*In 1975 collections, ANOVA was performed
separately and used this design.
on the upper and lower crown data
73
-------
TABLE 4.8. THE STRUCTURE OF THE FOUR-LEVEL NESTED ANALYSIS OF VARIANCE FOR BOTH 1975 AND 1976
COLLECTIONS, UPPER CROWNS.
Level 4 - Collections 1975 Collection 1976 Collection
Level 3 - Between the
Plots within Collections Plot 1 Plot 2 Plot 1 Plot 2
Level 2 - Between Tree
Age within the Plots Younger Older Younger Older Younger Older Younger Older
Level 1 - Between the
Ages of Foliage within
the Tree Ages Z4. ?3 72 747372 747372 747372 757473 757473 757473 757473
Level 0 - Five Individual 1 1 1
Observations for Each
Variable within the
Ages of Foliage
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
^
0
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
1
2
3
4
5
-------
the plots within the collections, at Level 2 are the two tree ages within the
plots, at Level 1 are the years of foliage, and at Level 0 are the five variates
for each year of foliage.
The results of the 1975 and 1976 fluoride analyses are shown in Table 4.9.
The F ratios for Level 4 (Between Collections) and Level 3 (Between the Plots
within the Collections) are significant at p ^ .05. It may be concluded that
mean values for fluoride from the two collections would be sufficiently
different such that they would represent samples from different populations;
similarly fluoride values from the plots represent samples from different
populations. Any level which showed effects significant at p ^ .05 were
further interrogated by t-tests.
HISTOLOGICAL STUDIES
Six plates of photomicrographs depicting three of the needle tissue
pathologies being studied on ponderosa pine foliage were prepared for this
report. Other pine foliage pathologies caused by insects are presented by Dr.
Bromenshenk in his portion of this report.
Needle Mottling
Air pollution studies have demonstrated that the macroscopic symptoms of
conifer needle mottling are a typical host manifestation to phytotoxic gases.
However, mottling of ponderosa pine needles can and does occur in pristine
areas where no or very low concentrations of phytotoxic gases are present. The
tissue pathology of mottling is first manifested in the mesophyll cell beneath
the rows of stomatal opening in localized areas of the needles. As illustrated
in Plate 1, which shows both cross and longitudinal sections of mottled pine
needles, the cell walls of the mesophyll cells beneath the stomatal openings
are destroyed in some cases and the surrounding cells then lose their
chlorophyll (photo D, Plate 1). In other cases, the mesophyll cells beneath
the stomatal openings lose their chlorophyll but retain intact cell walls which
lose none of their form (photo B, Plate 1).
Histological studies of mottled needles from the ponderosa sites in the
Colstrip area have not established any association thus far with either fungal
or insect infestation. Consequently, it appears that the needle mottling
recorded in this study is being caused primarily by abiotic factors such as
heat (i.e., concentration of the sun's rays by water droplets along stomatal
rows).
Basal Necrosis and Scale
During the 1974 and 1975 study periods, there was a strong possibility
that needle tissue pathologies occurring beneath or at the apical opening of
the fascicular sheath were caused by both biotic (insects) and abiotic (acidic
solutions) causal agents because there were two major macroscopic symptoms
involved with these pathologies. These two macrosymptoms were classified as
basal needle necrosis and basal needle scale. While both symptoms are
associated with necrotic tissues, basal scale is associated only with tissue
swelling (hypertrophy) and, in many cases, with the splitting of the epidermal
tissues on the swollen area of the needle. Although the authors are not
75
-------
TABLE 4.9. ANALYSIS OF VARIANCE TABLE FOR FLUORIDE IN BOTH 1975 AND 1976 COLLECTIONS, UPPER CROWNS.
Source of Variation
Sums of Squares Degrees of Freedom Mean Squares F Ratio
Between Collections
(Level 4)
Between the Plots
within the Collections
(Level 3)
Between the Tree Ages
within the Plots
(Level 2)
Between the Years of Foliage
within the Tree Ages
(Level 1)
Within the Years of Foliage
(Level 0)
23.914
25.314
5.340
12.335
94.360
10
40
240
23.913
3.164
.534
.308
.393
7.557*
5.925*
1.731
.784
*F ratios are significant @ p S .05
-------
-------
completely convinced that some insects may not cause tissue damage similar to
that classified as basal needle scale, histological studies of both tissue
necroses during 1976 and early 1977 strongly indicate that the two pathologies
are caused by abiotic agents.
Photomicrographs were prepared to depict the damage associated with basal
needle necrosis (Plates 2 to 4). On Plate 2 are three photomicrographs of a
cross section of the needle base and sheath of ponderosa pine collected from
the McRae pine site (SE-2). In photo B of this plate, one notes that there is
no endodermal tissue (see circled area on photo) on the dorsal side of the
needle where tissue necrosis occurred. This means that the causal agent
induced this damage before the parenchymatous cells in this area differentiated
into endodermal cells during early morphogenesis of the needle. Needle
morphogenesis of ponderosa pine takes approximately three months after the
spring bud break-candle stage period, and thus it is during this time that a
needle is damaged by the causal agent.
Both saprophytic fungi and small insects (see Bromenshenk's 1977 report on
insects) are common residents of the fascicular sheath area. In all four
photographs on Plate 2 one notes the saprophytic fungus Phylctaena sp. on the
outer cataphylls of the fascicular sheath. This saprophyte, which produces a
pycnidial fruiting body (see arrow in photo D), is the most common fungus found
in these histological studies of needle basal tissues. Since its hyphal
filaments (vegetative structure) are not found in either the necrotic or
healthy tissues of the pine needle, it is suspected that this saprophyte is
restricted to the dead sclerenchymatous cells of the fascicular sheath.
On Plate 3 are photomicrographs which depict cross sections of healthy
(photos A and B) and necrotic (photos C and D) basal needles collected from the
Kluver pine site (E-l) during 1976. The structure in the middle of the three
needle set is the dwarf shoot bud which usually remains dormant during the
entire life of the pine needles. However, depending on its positioning along
the internode, the dwarf shoot bud can initiate growth and form the male or
female cones. Also, dwarf shoot buds can form lateral branches if the terminal
bud of the branch is destroyed.
Photos A and B of Plate 3 depict the healthy tissue of a needle lacking any
basal tissue or cell necrosis. Photos C and D depict an interfacial (between
the needle) necrosis of the epidermal and hypodermal tissue area of all three
needles of this needle bundle (see arrow in photo C). This necrosis occurred
during early needle morphogenesis, since the epidermal and hypodermal cells had
not differentiated from the thin-walled parenchymatous cells. Therefore, the
causal agent(s) of the damage entered this area at an early stage of needle
tissue development. To facilitate understanding of this interfacial
epidermal-hypodermal tissue necrosis, Plate 4 was prepared showing a
peridermal longitudinal section of a ponderosa pine needle (Site E-l)
demonstrating basal tissue necrosis of the interfacial area of the needle. As
depicted (see arrows) in photos A, B, and C, the necrosis occurs in all the
hypodermal cells from the dwarf shoot bud to the outer periphery where one
notes the epidermal one-cell layer. When necrosis occurs in the hypodermal-
epidermal tissues of the needle, easy entry to the mesophyll tissue is
available to both parasitic or saprophytic fungi. However, this rarely occurs
78
-------
B
-------
PLATE 3
-------
and there is an extreme dearth of either type of fungi in needles manifesting
basal needle necrosis.
/
One notes in photos A, C, and D of Plate 4 that the dwarf shoot bud is
composed entirely of thin-walled parenchymatous cells and that no cell necrosis
is occurring in the apical portion of this bud (photo D shows a different
section of the same bud). Furthermore, one notes that cellular necrosis is
occurring at the base of the dwarf shoot bud, which strongly suggests that the
causal agent in this case is lodged only at the base of the bud during the
period of penetration for these cells. The photographs on both Plate 3 and
Plate 4 strongly support our theory that abiotic causal agents are responsible
for basal needle necrosis.
On Plate 5 are photomicrographs of cross sections of basal needle areas
manifesting various stages of basal scale. In photo A, the swelling beneath
the necrotic epidermal and hypodermal tissues is caused by the production of
excessive parenchymatous cells (hyperplasia); if these cells had developed
normally during early needle morphogenesis they would have differentiated into
mesophyl1 cells containing chloroplasts. In photo B, the needle cross section
depicts no external tissue necrosis but is manifesting the hypertrophy of
mesophyll cells and the symptoms of basal scale. One notes that the mesophyll
cells adjacent to the hypodermis (see arrow) are undergoing cell enlargement
(hypertrophy) but not hyperplasia. In both photos C and D, excessive
hypertrophy of mesophyll cells has caused the splitting of the hypodermal and
epidermal layers in the basal needle area.
The last plate (Plate 6) depicts cross sections of a ponderosa pine needle
manifesting basal necrosis (photos A and B) and a longitudinal section of a
ponderosa pine needle manifesting basal scale (photos C and D). As can easily
be discerned from these two tissue pathologies, the necrotic cells of the
"basal necrosis" symptom do not undergo hypertrophy prior to dying (photos A
and B), while hypertrophy of mesophyll cells does occur prior to death in the
"basal scale" symptom. Photos A and B of this plate also demonstrate (as
photos A and B, Plate 2) that the causal agent caused the damage to the basal
needle tissues before formation of the endodermal cells in the area of damage
(see arrow in photo B).
1975 DATA ANALYSIS
As previously mentioned in the introduction section of this reportK each
of the 12 pine foliage characteristics being measured is expected or
hypothesized to change if and when the atmospheric emissions of coal-fired
power plants impact the permanent ponderosa pine-skunkbush sites. The results
of the 1975 ponderosa pine-skunkbush studies are presented first so that the
reader realizes where the similarities and significant differences within the
12 characteristics occurred before any coal-fired power plants went on-line at
Colstrip. After the 1975 data, the results of the 1976 data are presented, and
the variability between the 1976 data and 1975 data are then compared utilizing
four-level Analysis of Variance (ANOVA).
The data for the 12 growth/health/disease characteristics from both crown
positions and the upper and lower crown positions separately from 1975 were
81
-------
; "v*"' : Si
.'&
i&w^^jwiiTO" wii
^^MP^-^:"
•
PLATE 4
82
-------
PLATE 5
83
-------
PLATE 6
84
-------
interrogated by nested ANOVA. These ANOVA designs were previously outlined
under Materials and Methods.
The F ratios and the percent variance components for all 12 characteris-
tics from the four-level nested ANOVA, employing both crown positions, are
shown on Table 4.10. The F ratios and percent variance components from the
ANOVA for upper crown positions are shown on Table 4.11, and those for lower
crown positions are arrayed in Table 4.12. The data on these three tables are
summarized in Table 4.13, and significant treatment effects (p ^ .05) of any of
the 12 characteristics are indicated by asterisks (*).
Inspection of Table 4.10 shows that all growth/health/disease character-
istics have significant differences between the plots except for cross-
sectional area. The cross-sectional area, however, was the only one which
showed a significant treatment effect between the crown positions within the
plots (Table 4.10, Level 3). Sulfur levels, cross-sectional area, percent
total necrosis, and percent mottled needles showed significant treatment
effects due to tree age within the two crown positions (Table 4.10, Level 2).
Percent needle retention, both basal needle pathologies, percent tip burn, and
percent healthy needles showed significant treatment effects between the three
different years of foliage within given tree ages (Table 4.10, Level 1).
As mentioned, Table 4.11 shows the F ratios and percent variance
components for 1975 data from the upper crown. Seven of the 12 measured
characteristics (i.e., fluoride, cross-sectional area, needle length, percent
basal necrosis, percent defoliator, percent tip burn, percent mottled needles)
for the upper crown position show significant treatment effects between the
five plots (Table 4.11).
Treatment effects due to tree age were found for sulfur, percent total
necrosis, and percent mottled needles in the upper crown foliage as was found
in the ANOVA for both crown positions (Table 4.10) but not for needle cross-
sectional area (Table 4.10, Level 2). Significant treatment effects due to
year of foliage of the upper crown are present for percent basal scale, percent
basal necrosis, percent tip burn, and percent healthy needles, but not for
percent needle retention as was found in the ANOVA using both crown positions
(Table 4.10, Level 1).
Table 4.12 contains data obtained from three-level nested ANOVA of the 12
growth/health/disease characteristics of foliage for the 1975 data from the
lower crown position. Five of the seven foliage characteristics which had
significant treatment effects in the upper crown between plots (Table 4.11,
Level 3) showed significant treatment effects in the lower crown; cross-
sectional area and percent basal necrosis were not significant at this level
(Level 3). A treatment effect due to tree age (Table 4.12, Level 2) was present
only for needle cross-sectional area. Percent needle retention, the two basal
needle pathologies, percent tip burn, and percent healthy needles showed
significant treatment effects due to years of foliage (Table 4.12, level 1) as
was found in the ANOVA using both crown positions (Table 4.10, Level 1). The
data on T-ables 4.10, 4.11, and 4.12 at Level 0 demonstrate that more than 50
percent of the total variance was found within the individual year's foliage.
85
-------
CO
TABLE 4.10. SUMMARY OF F RATIOS AND % VARIANCE COMPONENTS FOR THE INDICATED LEVELS FOR ALL VARIABLES
FROM THE 1975 COLLECTION, BOTH CROWN POSITIONS.
Level 4
Between
Plots
Variable
Fluoride (ppm)
Sulfur (ppm)
Area (mm2)
Needle length (mm)
% Needle
Retention
% Basal Necrosis
% Basal Scale
% Defoliator
Percent
F Variance
Ratio Component
15.0313*
15.6357*
.3812
8.1780*
6.0267*
6.4512*
25.1439*
26.5435*
% Tip Burn 559.6725*
% Total Necrosis
% Mottled Needles
% Healthy Needles
6.2189*
14.0596*
6.3088*
20.3
23.7
-12.7
28.1
5.5
1.0
7.0
37.2
35.9
10.1
30.9
-5.5
Level 3
Between
Crown Positions
Within Plots
Percent
F Variance
Ratio Component
.9893
.4764
6.4182*
2.7015
.7143
2.2886
.2011
.7619
.0407
.2298
1 . 2688
.2325
.17
-5.6
19.2
5.3
1.7
9.3
2.5
-.77
-1.0
-7.0
-1.7
7.3
Level 2
Between Tree Ages
Within
Crown Position
F
Ratio
1.0325
4.0962*
3.0008*
1.2374
.7599
.1746
.5748
1.5119
.8610
2.3471*
4.5441*
.2533
Percent
Variance
Component
.18
8.2
5.4
1.1
-2.4
-12.8
-6.0
2.5
-1.0
9.0
5.3
-13.7
Level 1
Between Years
Fol iage Within
Tree Ages
Percent
F Variance
Ratio Component
1.0912
.4883
.4092
1.1608
1.8939*
3.3341*
3.1461*
1.2710
2.0059*
1.1983
.2996
3.8790*
1.4
-8.3
-11.7
2.0
14.4
32.5
28.9
3.1
11.0
3.3
10.6
40.9
Level 0
Residual
Percent
Variance
Component
77.9
82
99.8
63.3
80.7
69.8
67.4
57.9
55.0
84.4
76.0
71.0
*F ratios significant at p ^ .05
-------
TABLE 4.11. SUMMARY OF F RATIOS AND % VARIANCE COMPONENTS FOR THE INDICATED LEVELS FOR ALL VARIABLES
FROM THE 1975 COLLECTIONS, UPPER CROWN POSITIONS.
Level
3
Between
Plots
Variable
Fluoride (ppm)
Sulfur (ppm)
Area (mm2)
Needle Length (mm)
% Needle Retention
% Basal Necrosis
% Basal Scale
% Defoliator
% Tip Burn
% Total Necrosis
% Mottled Needles
% Healthy Needles
F
Ratio
9.
3.
5.
10.
1.
17.
3.
8.
7.
6.
1.
.1449*
,8730
,6601*
.4802*
,4999
,2936*
.6641
.9789*
8807*
6175
5830*
5612
Level 2
Level 1
Between
Tree Ages
Within Plots
Percent
Variance
Component
27
12
8
32
3
18
11
33
34
-6
26
7
.5
.9
.6
.9
.5
.8
.1
.4
.8
.9
.5
.4
F
Ratio
1.9399
7.6885*
1 . 3488
1.2172
1.5102
.0862
.2469
1.9726
1.4351
3.6924*
6.8488*
.2442
Percent
Variance
Component
3.
10.
1.
1.
4.
-14
-10
4.
2.
16.
9.
-10.
4
2
1
2
1
0
9
0
1
8
Between
Fol iage
Tree
F
Ratio
.7511
.2550
.5007
1 . 3740
1.4177
3.7867*
2.9838*
1.0508
1.9609*
.9812
.3136
2.8365*
Years
Within
Ages
Percent
Variance
Component
-3.
-13.
-10
4.
7.
34
28.
•
10
-3.
-10.
27.
6
4
5
1
2
6
4
2
7
Level 0
Within
Years
Foliage
Percent
Variance
Component
72.6
90.2
100
61.2
85.2
61.6
71.2
61.7
52.1
91.2
74.5
75.6
*F ratios significant at p g .05
-------
TABLE 4.12. SUMMARY OF F RATIOS AND % VARIANCE COMPONENTS FOR THE INDICATED LEVELS FOR ALL VARIABLES
FROM THE 1975 COLLECTIONS, LOWER CROWN POSITIONS.
Variable
Fluoride (ppm)
Sulfur (ppm)
Area (mm2)
oo Needle Length (mm)
% Needle Retention
% Basal Necrosis
% Basal Scale
% Defoliator
% Tip Burn
% Total Necrosis
% Mottled Needles
% Healthy Needles
F
Ratio
6.
4.
3.
13.
4.
4.
2.
11.
20.
1.
29.
•
4123
2540
1453
7680
6608
4609
3684
9581
1396
0794
0267
4061
Level 3
Between
Plots
Level 2
Level 1
Between
Tree Ages
Within Plots
Percent
Variance
Component
* 14
16
9
* 32
11
12
13
* 36
* 35
* 27
9
.3
.2
.8
.3
.8
.2
.9
.0
.54
.1
.0
F
Ratio
.6068
2.3751
6.9123
1.2750
.2831
.3060
.9208
1.1603
.4319
1.7427
1.3147
.2599
Percent
Variance
Component
-2.
6.
* 10.
1.
-8.
-9.
-1.
-4.
5.
•
-14.
9
3
5
1
7
2
0
9
4
7
49
8
Between
Fol iage
Tree
F
Ratio
1 . 3845
.8698
.2856
.9044
2.4079*
2.8310*
3.3777*
1.5131
2.0409*
1.3306
.2819
5.2829*
Years
Withi
Ages
n
Percent
Variance
Component
6.
-2
-13.
-1.
21.
25.
27.
5.
11.
5.
-12.
48.
3
3
2
4
8
9
7
9
8
1
8
Level 0
Within
Years
Foliage
Percent
Variance
Component
82.5
79.3
93.6
67.3
76.0
70.5
59.8
56.3
57.4
87.8
84.4
56.9
*F ratios significant at p g .05
-------
TABLE 4.13. SUMMARY OF ANALYSES OF VARIANCE FOR 1975 COLLECTION.
Between the Tree Ages
Between the Plots Within the Plots Between the Years of Foliage
(Level 4) (Level 3) (Level 3) (Level 3) (Level 2) (Level 2) (Level 2) (Level 1) (Level 1)
Both Upper Lower Bothtt Upperttt Lowerttf Both Upper Lower
Crown Crown Crown Crown Crown Crown Crown Crown Crown
Variable Positions Position Position Positions Position Position Positions Position Position
Fluoride * * *
(ppm)
Sulfur (ppm) * * *
Area (mm2)t * * *
Needle * * *
Length (mm)
% Needle * * *
Retention
"• % Basal
Necrosis
% Basal
Scale
% Defoliator * * *
% Tip Burn * * *
% Total *
Necrosi s
% Mottled * * *
Needles
% Healthy *
Needles
t This variable showed significant differences between the Crown Positions
tt This level is actually Between the Tree Ages Within the Crown Positions
ttt This level is Between the Tree Ages Within the Plots
Significant treatment effect at p ^ .05
*
-------
Fluoride
The only significant effect for fluoride in the three-or four-level nested
ANOVA was between the plots for all crown positions as shown in Table 4.13. No
significant treatment effects for fluoride were detected between tree ages or
between years of foliage. T-tests were performed between the categories of
tree age and crown positions for fluoride data for.foliage from all plots to
confirm this, and the t-statistics for plot SE-2 are presented in Figure 4.1
(plot SE-2 was representative of all five plots for fluoride). Figure 4.2 was
prepared to show the mean values for fluoride in 1974 foliage for all upper
crown samples from all five plots, and this figure illustrates the difference
in mean values between the plots. The mean fluoride values shown in Figure 4.2
between plots SE-4 and those of E-l, SE-2, and S-5 were significantly different
at p ^ .05.
Sulfur
Significant treatment effects between the plots and between the tree ages
for sulfur levels were detected (Table 4.10, Levels 2 and 4) by ANOVA employing
both crown positions and by ANOVA employing the upper crown position (Table
4.11, Level 2), which suggests greater variation between upper crowns of the
two tree ages than between the lower crown positions.
T-tests of sulfur levels between the individual years of foliage from
upper crowns of both older trees and younger trees, tested separately, revealed
no significant differences (p S .05). Thus, the mean values of sulfur
concentrations for upper crown younger trees and upper crown older trees were
recomputed by pooling the measurements for the individual years. These means
for upper crown younger trees and upper crown older trees were then compared
utilizing the t-test. Plot SE-4 was the only plot which showed significant
differences for sulfur between the tree ages at p ^ .05, and the basic
statistics are presented in Table 4.14. The means and 95% confidence intervals
for sulfur levels between upper crown, younger and older trees are shown in
Figure 4.3 for plot S-5.
i
Cross-Sectional Area
For needle fascicular cross-sectional area, ANOVA indicated significant
treatment effects (Table 4.13) between the plots for the upper crown position.
Also, significant effects were found between the tree ages for the lo^er and
for the combined crown positions (see Table 4.13) and between the crown
positions for both crown analyses. It should be expected that the variability
of cross-sectional area of fascicles from lower crown foliage within the plots
will be greater than between the plots. Figure 4.4 illustrates the mean values
and 95% confidence intervals for fascicular cross-sectional areas from the
upper and lower crown positions for plot SE-4. As is depicted in Figure 4.4,
the difference between the crown positions was most apparent for the younger
trees and fascicular cross-sectional area was generally smaller for the lower
crown position.
90
-------
1974
UO
LY
LO
UY
DO
LY
.416
.162
.461
1.068
1.473
.913
XY
UZ
ZO LZ
.625
1.163
1973
UO
LY
LO
UY
UO
LY
.035
.069
.063
.382
.654
.698
ZY
UZ
ZO LZ
.388
.328
UO
LY
LO
1972
UY
UO
LY
.100
1.343
1.921
.149
.331
1.572
ZY
UZ
ZO LZ
1.023
1.279
*t.05 = 2.306 for UY, UO, LY, LO Comparisons
t.05 = 2.101 for ZY, ZO, UZ, LZ Comparisons
**UY = Upper Crown, Younger Trees ZY
UO = Upper Crown, Older Trees ZO
LY = Lower Crown, Younger Trees UZ
LO = Lower Crown, Older Trees LZ
All Crowns, Younger Trees
All Crowns, Older Trees
Upper Crowns, All Tree Ages
Lower Crown, All Tree Ages
Figure 4.1. Results of t-tests* between the categories** of tree age and
crown position for 1974, 1973, and 1972 years of foliage from
plot SE-2.
91
-------
PLOT SE-4
-| PLOT S-5
PLOT E-l
ro
^ PLOT SE-2
^ PLOT S-3
PPM FLUORIDE
Figure 4.2. Mean values and 95% confidence interval widths for fluoride in 1974 foliage, upper crown,
al1 tree ages.
-------
1974
-| 1973 OLDER TREES
1 1974
1973
1972
YOUNG TREES
I
1
200
1 ft I 1972
I " ' • 1 YOUNG TREES*
g ^^ OLDER TREES'1
*ALL YEARS FOLIAGE COMBINED
1 1 ' 1 1 1 ' 1 ' 1 ' 1 ' 1 '
300 400 500 600 700 800 900
PPM SULFUR
Figure 4.3. Variation in total sulfur concentration in upper crown younger and older trees for plot S-5.
-------
| — + - 1 1974
YOUNGER TREES
1973 UPPER CROWN
1972
1974
1973
LOWER CROWN
— | 1972
to
OLDER TREES
t-H
UPPER CROWN
LOWER CROWN
.5
I I I I I
1.0 1.5 2.0 2.5 3.0
FASCICULAR CROSS SECTIONAL AREA, nm2
I
3.5
Figure 4.4. Mean values and 95% confidence intervals for fascicular cross-sectional area, upper and
lower crown positions, younger and older trees, plot SE-4.
-------
TABLE 4.14. BASIC STATISTICS FOR SULFUR CONCENTRATIONS IN UPPER CROWN YOUNGER
TREES AND UPPER CROWN OLDER TREES, ALL YEARS FOLIAGE COMBINED,
PLOT SE-4*
Statistic Upper Crown, Younger Upper Crown, Older
N
Maximum Value
Minimum Value
Mean
Variance
Standard Deviation
Standard Error
95% Confidence Limit
Median
Standard Error
Coefficient of Variation
Standard Error
Sampling Precision**
15
550
300
420
5285.714
72.03
18.772
40.266
450
23.5
17.31
3.25
9.587
15
500
250
348
6291.666
79.320
20.480
43.930
350
25.666
22.771
4.36
12.612
*t-statistic @ p £ .05 = 2.048
t-statistic between the mean values = 2.580
**Defined as
100
Needle Length
Significant treatment effects were found for needle length between plots
but not within plots (Table 4.13). The mean values for needle length for each
year's foliage for upper and lower crown, younger and older trees for each plot
were compared by t-tests and the results are summarized in Table 4.15. As is
depicted in this table, needle length across the plots was generally shorter on
the lower crowns than on the upper crowns of younger trees, while needle length
of older trees of both upper and lower crowns was generally the same. The upper
crowns of younger trees tended to have longer needles than the upper crowns of
older trees.
Needle Retention
ANOVA showed significant treatment effects for needle retention between
the plots (Table 4.13) and between the years of foliage for both crown positions
and the lower crown position but not for the upper crown position. The mean
values and 95% confidence intervals for percent needle retention for upper and
lower crown positions for plot E-l are shown in Figure 4.5 and these values are
generally representative of the other plots. The obvious differences between
1974 percent needle retention and older foliage from the lower crown as depicted
in Figure 4.6 is representative of the patterns at the other plots with the
exception of plot S-3 where the percent needle retention on the lower crowns was
95
-------
TABLE 4.15. RESULTS OF T-TESTS BETWEEN MEAN VALUES FOR NEEDLE LENGTH FOR
TREE AGES AND CROWN POSITIONS FOR ALL PLOTS.
Plot
SE-4
S-5
E-l
SE-2
S-3
Year of
Foliage Origin
1974
1973
1972
1974
1973
1972
1974
1973
1972
1974
1973
1972
1974
1973
1972
Mean
UY* vs
110.7
127.6
117.3
128.3
150.2
146.1
151.0
160.8
160.2
134.1
145.7
130.5
130.1
138.0
120.3
Values for
UO*
131. 2t
124.6
1 11 . 5t
115. 5t
141.5t
136. 7f
152.9
153. 3t
145.91
119. 9f
137. 5t
135. 2t
119.01
123. 5t
1 1 1 . 4t
Needle
LY*
126.6
118.1
111.6
120.0
127.5
122.5
152.7
144.1
145.1
127.3
125.3
121.8
127.8
126.8
116.9
Length
vs. LO*
135. 9f
125. Of
111.7
122.5
125.4
110.3f
146. 5f
151.lt
140.4
122. 3f
138.lt
128.2f
113. 7f
114. 6f
110. 4t
Plot
SE-4
S-5
E-l
SE-2
S-3
Year of
Fol iage Origin
1974
1973
1972
1974
1973
1972
1974
1973
1972
1974
1973
1972
1974
1973
1972
Mean
UY* vs
110.7
127.6
117.3
128.3
150.2
146.1
151.0
160.8
160.2
134.1
145.6
130.5
130.1
138.0
120.3
Values for
LY*
126. 6t
118.lt
111.6t
120. Of
127. 5t
122. 5t
152.7
144.lt
145. If
127. 3f
125. 3f
121. 8f
127. 8f
126. 8f
116.9
Needle
UO*
131.2
124.6
111.5
115.5
141.5
136.7
152.9
153.3
145.9
119.9
137.5
135.2
119.0
123.5
111.5
Length
vs. LO*
135.9
125.0
111.7
122. 5f
125. 4f
110. 3f
146.5
151.0
140.4
122.3
>38.1
128.2f
113. 7t
114. 6f
110.4
*UY = Upper Crown, Younger Trees LY = Lower Crown, Younger Trees
UO = Upper Crown, Older Trees LO = Lower Crown, Older Trees
fMean values are significantly different @ p ^ .05
96
-------
•\ 1974
| 1973
1972
UPPER CROWN
UD
•-J
-\ 1973
1974
LOWER CROWN
1972
I
50
I
60
70
I
80
i
90
I
100
PERCENT NEEDLE RETENTION
Figure 4.5. Mean values and 95% confidence intervals for percent needle retention for upper and lower-
crown positions, plot E-l.
-------
PLOT
SE-4
1974
1973
1 1972
PLOT
S-5
^ 1973
-\ 1974
1972
PLOT
E-l
1972
^ 1974
1973
PLOT
SE-2
1972
1974
1973
PLOT
S-3
•4-
1973
1972
1974
I I I i
20 40 60 80
PERCENT NEEDLE RETENTION
100
Figure 4.6. Mean values and 95% confidence intervals for percent needle
retention for upper crown positions for all plots.
98
-------
90 percent for all internodes. At all plots, mean percent needle retention de-
creases with internode age for upper crown positions, but the differences were
usually significant only between the youngest internodes (i.e., 1974 foliage)
and the oldest internodes (i.e., 1972 foliage) being examined.
Basal Necrosis
As depicted in Table 4.13, significant treatment effects for basal necrosis
were found between the plots and between the years of foliage for both crown
positions and for the upper crown position, while significant treatment effects
in the lower crown position were found only between the years of foliage. The
mean values for percent basal needle necrosis and the 95% confidence intervals
for upper and lower crown positions for plots E-l and SE-2 are shown in Figure
4.7. As is depicted in this figure, the mean values for percent basal necrosis
increased with internode age in general, but differences between the mean values
were not always significant across the plots.
Basal Scale
The ANOVA of basal scale showed that there were significant treatment
effects in this pathology for both crown positions between the plots and between
the years of foliage. For the upper and lower crown positions, significant
treatment effects were indicated between the different years of foliage (see
Table 4.13). No significant treatment effects were detected for basal scale due
to tree age. Percent basal scale is quite variable, and the mean values are
imprecisely known. On Table 4.16 are the basic statistics for percent basal
scale for the three different-aged foliage from plot S-3, and these data are
illustrative of the variability of the data from the other plots.
Defoliator
For percent defoliator, the ANOVA detected significant treatment effects
between the plots for both crown positions and for lower and upper crown
positions, but not between the tree ages nor between the years of foliage, which
indicates that the variability for this pathology was greater between the plots
than within the plots (see Table 4.13). As was found for basal scale, percent
defoliator was a very variable needle pathology. The mean values and 95%
confidence intervals for all plots, upper and lower crown positions, are arrayed
in Table 4.17.
Tip Burn
In Table 4.13, significant treatment effects between the plots and between
the different years of foliage were detected for percent tip burn by each ANOVA.
No significant effects were found between the younger and older trees or between
crown positions. The basic statistics for percent tip burn for the three
different-aged foliage from plots S-3 and SE-2 are presented in Table 4.18.
99
-------
1974
1973
1972
1974
1973
1972
1974
1973
1972
1974
, UPPER CROWN
' 1
I t t
1 * 1
H
1, 1 LOWER CROWN
1 ' 1
1 t 1
1 * 1
J UPPER CROWN
iJI
^ PLOT
SE-2
H
1973 JL 1
1972
PLOT
E-l
r -
0
i
5
I
10
l
15
i
20
i
25
PERCENT BASAL NECROSIS
Figure 4.7. Percent basal necrosis for upper and lower crown positions,
plots E-l and SE-2, illustrating differences between internodes
and between plots.
100
-------
TABLE 4.16.
BASIC STATISTICS FOR PERCENT BASAL SCALE FOR 1974, 1973, AND 1972
YEARS FOLIAGE FROM PLOT S-3.
1974
1973
1972
N
Maximum Value
Minimum Value
Mean
Standard Deviation
95% Confidence Intervals
Median
Standard Error
Coefficient of Variation
Standard Error
20
12
0
6.
+ 1.
1.
125.
40.
69
00
05
57
251
682
1
25
20
37
0
8.
10.
+5.
-4.
8.
3.
65.
14.
23
94
55
22
49
06
66
17
20
54
0
12.
13.
+8.
-6.
10
3.
66.
14.
.52
.72
29
42
84
22
34
TABLE 4.17. MEAN VALUES AND 95% CONFIDENCE INTERVALS FOR PERCENT DEFOLIATOR
FOR UPPER AND LOWER CROWN POSITIONS, ALL PLOTS.
1974
Upper Crown
1973
1972
Plot
SE-4
S-5
E-l
SE-2
S-3
0
.44
2.93
2.89
3.41
1.3
3.67
3.03
5.44
.44
2.28
1.97
2.93
1.
4.
2.
5.
11
44
41
59
99
.43
2.0 1.
3.70 2.
3.23 1.
4.14 3.
11
15
62
59
10
.02
1.20
7.51
8.41
8.18
.19
1.62
11.12
4.19
10.19
6.
3.
6.
,01
.94
.34
.40
53
Lower Crown
1974
1973
1972
Plot
SE-4
S-5
E-l
SE-2
S-3
.07
1.89
1.98
2.06
6.22
.42
2.16
4.78
2.79
6.38
.05
1.35
1.94
1.63
4.24
.04
2.74
5.96
6.37
7.85
.21
2.64
7.98
4.77
5.43
.03
1.77
4.75
3.51
4.10
.11
4.10
9.09
8.68
9.08
.63
3.45
4.58
4.38
6.74
.08
2.43
3.72
3.56
5.01
101
-------
TABLE 4.18. BASIC STATISTICS FOR % TIP BURN FOR THE INDICATED YEARS FOLIAGE
FOR PLOTS SE-2 AND PLOTS S-3.
Statistic
N
Maximum Value
Minimum Value
Mean
Variance
Standard Deviation
SE-2*
1974 1973
20 20
12 52
0 0
2.01 10.5
50.9 200
7.13 14.1
95% Confidence Interval +1.95 +8.0
-1.30 -5.98
Median
1.45 5.0
95% Confidence Interval +2.26 +8.0
-1.22 -4.35
Coefficient of Variation 87.59 74.6
Standard Error
22.0 17.1
Sampling Precision** 81.2 66.6
S-3*
1972 1974 1973
20 20 20
73 4 6
000
23.2 .08 .03
335 12.7 11.21
18.3 3.56 3.35
+13.64 +.25 +.017
-11.26 -.08 -.020
25.9 ' 0 0
+17.7 +.13 +.11
-14.4 -.13 -.11
63.5 212.8 337.0
13.5 106.7 259.6
53.6 199 315.3
1972
20
2
0
.07
7.87
2.8
+ .17
-.07
0
+ .08
-.08
180.4
78.2
168.8
*The data arrayed
position combinec
**Defined as (t
X
herein are for all categories of tree age and crown
5Sx-* 100
102
-------
Total Needle Necrosis
ANOVA showed significant treatment effects between the plots and between
the tree ages for both crown positions (Table 4.13). Significant effects were
detected for the different-aged trees for upper crown but not lower crown. No
significant effects of percent total needle necrosis were found between the
crown positions or between the different years of foliage. The mean values and
the 95% confidence intervals for percent needle necrosis for SE-2 are presented
in Figure 4.8.
Needle Mottling
As shown in Table 4.13, the ANOVA detected significant treatment effects
in percent mottled needles between the plots for the three categories of crown
positions, and in the two tree ages for both crown positions and the upper
crown position. No differences were detected due to years of foliage origin.
The mean values and 95% confidence intervals for the foliage of upper crowns of
younger and older trees from plots SE-2 and S-3 are depicted in Figure 4.9.
Heal thy Needles
Significant treatment effects were detected between plots for the ANOVA
employing both crown positions and for all three combinations of crown
positions between the years of foliage (see Table 4.13). Mean values for
percent healthy needles and the 95% confidence intervals for the upper crown
positions for all five ponderosa sites are depicted in Figure 4.10.
1975-1976 DATA ANALYSES COMPARISON
The data from the 1976 collections were treated by a three-level nested
ANOVA, and the results are summarized as Table 4.19. In addition, the data for
foliage from the upper crown positions between collections for both years were
treated by four-level nested ANOVA, and the results are shown in Table 4.20.
Tables 4.11, 4.19, and 4.20 are summarized in Table 4.21. In this table the
four-level nested ANOVA between the 1975 and 1976 collections is designated
ANOVA K, the three-level nested ANOVA for upper crown foliage from the 1975
collections is designated ANOVA B, and the three-level nested ANOVA for the
1976 collection is designated ANOVA J. For each variable in Table 4.21, a
significant treatment effect (p ^ .05) for a particular level, for each ANOVA
(K, B, or J, as above), is denoted by an asterisk (*). Then, for ppm fluoride
(refer to Table 4.21) significant treatment effects were detected: (1) Between
the two years' collections (Level 4, ANOVA K), and (2) between the plots (Level
3, ANOVAs K and B).
Significant treatment effects between the 1975 and 1976 collections
(Table 4.21, ANOVA K) were detected only for: (1) ppm fluoride, (2) percent
total necrosis, and (3) percent healthy needles. Significant treatment effects
only between the plots were indicated for area (mm2), needle length (mm), and
percent defoliator by each ANOVA. For percent basal scale, significant
103
-------
1974
1973
1972
1974
o
1973
1972
YOUNGER
OLDER
8 10 12
PERCENT TOTAL NECROSIS
14
I
16
18
20
Figure 4.8. Mean values and 95% confidence intervals for percent total necrosis illustrating differences
in mean values for upper crown, younger and older trees at plot SE-2.
-------
1974
1973
1972
o
Ul
1974
1973»-
1972
1974
1973
1972
1974 4.
1973>|
1972*-
YOUNGER TREES
PLOT
SE-2
OLDER TREES
YOUNGER TREES
PLOT
S-3
OLDER TREES
10
I
12
1
I
16
li
Figure 4.9.
PERCENT MOTTLED NEEDLES
Mean values and 95% confidence intervals for percent mottled needles illustrating
differences in mean values for upper crown, younger and older trees for plots SE-2 and S~3.
-------
PLOT
SE-4
I
1974
1 1973
1972
I
PLOT
S-5 I • 1 1973
1974
-\ 1972
PLOT
E-l
»
1972
^ 1974
1973
PLOT
SE-2 h
1972
» 1974
1973
PLOT
S-3
•*•
1973
1972
1974
20
I
40
60
PERCENT HEALTHY NEEDLES
80
I
100
Figure 4.10. Percent healthy needles for all plots, upper crown positions,
illustrating differences between internodes.
1Q6
-------
TABLE 4.19. SUMMARY OF F RATIOS AND % VARIANCE COMPONENTS FOR THE INDICATED LEVELS FOR ALL VARIABLES
FROM THE 1976 COLLECTIONS, UPPER CROWN POSITIONS.
Level
3
Between
Plots
Fl
Variable
uoride (ppm)
Sulfur (ppm)
Area (mm2)
Needle Length (mm)
%
%
-------
TABLE 4.20. SUMMARY OF F RATIOS AND % VARIANCE COMPONENTS FOR THE INDICATED LEVELS FOR ALL VARIABLES
FROM THE 1975 AND 1976 COLLECTIONS, UPPER CROWN POSITIONS.
o
CO
Level 4
Between
Col lections
Variable
Fluoride (ppm)
Sulfur (ppm)
Area (mm2)
Needle Length (mm)
% Needle
Retention
% Basal Necrosis
% Basal Scale
% Defoliator
% Tip Burn
% Total Necrosis
% Mottled Needles
% Healthy Needles 1
F
Ratio
7.5576*
2.2318
1.2522
.0479
.8059
2.7653
2.7453
.6829
.1085
5. 3700*
4.1453
1.2915*
Percent
Variance
Component
21.
6.
-.
-11.
-1.
3.
-1.
-5.
-10.
6.
11.
10.
45
06
80
99
28
57
65
85
38
97
19
21
5
7
6
10
1
15
1
8
7
1
19
6
Level 3
Between
Plots Within
Col lections
F
Ratio
.9252*
.2439*
.2892*
.3693*
.5374
.1392*
.5825
.1998*
.7407*
.4337
.5443*
.4417*
Percent
Variance
Component
13.65
27.74
14.20
35.68
5.55
24.34
13.01
31.03
32.04
5.68
24.19
15.52
Level 2
Between
Withi
F
Ratio
1.7318
5.5503*
1.2935
1.4814.
.8097
.2987
.3352
1.9918
.6565
1.0276
.4014
.1244
Tree Ages
n Plots
Percent
Variance
Component
2.49
7.99
1.30
2.49
-2.09
-6.87
-13.78
4.41
-4.21
.30
-3.63
-15.19
Level 1
Between Years
Foliage Within
Tree Ages
F
Ratio
.7843
.3939
.7410
1.0661
2.0411*
2.3314*
6.1854*
.9365
3.2129*
2.4593*
1.4591*
5.5676*
Percent
Variance
Component
-2.81
-8.12
-4.66
.96
16.85
16.58
52.14
-.90
25.32
19.66
5.73
42.70
Level 0
Within
Years
Foliage
Percent
Variance
Component
65.20
66.32
89.95
72.85
80.96
62.28
50.28
71.31
57.22
67.36
62.50
46.75
^Indicates F ratio significant at p ^ .05
-------
TABLE 4.21. SUMMARY OF RESULTS OF THREE-LEVEL NESTED ANALYSIS OF VARIANCE FOR UPPER CROWN FOLIAGE FROM
THE 1975 COLLECTIONS (ANOVA B), THE 1976 COLLECTIONS (ANOVA J), AND THE FOUR-LEVEL NESTED
ANALYSIS OF VARIANCE BETWEEN THE 1975 AND 1976 COLLECTIONS (ANOVA K).
Variable
Fluoride (ppm)
Sulfur (ppm)
Area (mm2)
MaaHIo I annth fmm^
Level 4
Between
Collections
ANOVA
K
*
Level 3
Between the Plots
ANOVA
K B J
* *
* *
* * *
* * *
Level 2
Between the
Tree Ages Within
the Plots
ANOVA
K B J
* * *
Level 1
Between the Years of
Foliage Within the
Tree Ages
ANOVA
K B J
% Needle Retention
% Basal Necrosis
% Basal Scale
% Defoliator
% Tip Burn
% Total Necrosis
% Mottled Needles
% Healthy Needles
* Indicates F ratio significant at p ^ .05
-------
treatment effects only between the years of foliage were detected by each
ANOVA, while for percent tip burn, significant treatment effects between the
plots and between the years of foliage were noted by each ANOVA (Table 4.21).
Since no significant treatment effects for tree ages or age of foliage
origin were detected, values for all tree ages and years of foliage were
combined, and the resultant mean values and 95% confidence intervals for
fluoride from upper crown positions from all plots, both collections, are shown
in Figure 4.11. Whereas in 1975 the plots were significantly different in
fluoride content, in general, in 1976, with the exception of plot SE-2, the
plots were not significantly different.
For ppm sulfur, significant treatment effects between the plots were
detected by ANOVAs K and J, but not for 1975 data alone (ANOVA B). ANOVAs B, K,
and J detected significant differences for sulfur between the tree ages. These
results indicate that changes in sulfur concentrations in 1976 were sufficient
to cause the plots themselves to be different, but the changes were not such
that the two year's collections would be different. The only statistically
significant difference between older and younger trees in 1976 was at plot S~5.
The basic statistics for sulfur concentrations in upper crown, younger and
older trees for plot S-5 are arrayed in Table 4.22.
The mean sulfur values for younger and older trees for all plots are
arrayed in matrix form as Table 4.23. For younger trees, plot SE-4 was
significantly lower than all other plots, while S-5 had significantly greater
concentrations than all other plots. Although plot E-l had more sulfur than
plot SE-4, it was lower than plots SE-2, S-3, and S-5. There were no signif-
icant differences between sulfur concentrations between plots S-3 and SE-2.
For older trees (Table 4.23), plot SE-4 was again significantly lower in sulfur
than all other plots. In addition to plot SE-4, plot S-5 had higher levels in
foliage than plot E-l. No other significant differences could be shown for
older trees between plots.
The mean values and 95% confidence intervals for sulfur in upper crown,
trees for 1975 and 1976 collections for each plot are shown in Figure 4.12. The
increases shown for plots S-5, SE-2, and S-3 in 1976 over 1975 are significant
at p ^ .05.
For percent needle retention, the ANOVA for 1975-1976 detected a signifi-
cant treatment effect between the years of foliage, as did the ANOVA £or the
1976 collection (Table 4.21). No significant effect for year of foliage'origin
was detected for the 1975 collection (Table 4.21, ANOVA B). The mean percent
needle retention and 95% confidence intervals for plot E-l, both collections,
are shown in Figure 4.13. The mean values for 1975 and 1974 years' foliage from
plot E-l are essentially the same, while that for 1973 year's foliage is nearly
12 percent lower. Whereas for the 1975 data, significant differences occurred
usually only between the 1974 and 1972 internodes, the 1976 data generally show
significant differences between 1973 and both 1974 and 1975 internodes.
For percent basal necrosis (Table 4.21) significant treatment effects
were shown for 1975-1976 and 1975 between the years of foliage, but not for the
1976 collection. The mean values and 95% confidence intervals for basal
110
-------
PLOT
SE-4
PLOT
SE-5
PLOT
E-l
PLOT
SE-2
PLOT
S-3
2 3
PPM FLUORIDE
Figuv
ure
kll. Mean values and 95% confidence
combined, for all plots. 1975 (-
intervals for fluoride
-) and 1976 ( — 4—)
in upper crowns
col lections.
all years foliage
-------
ro
1975
PLOT
SE-4
1976
1975
PLOT
S-5
1976
200
| • 1 1975 PLOT
E-l
1 * | 1071; PLOT
SE-2
I.., A I 1 07C
| • 1 1975 PLOT
S-3
1^ 1 1 r\-7 r
» I i y / u
1 1 1 1 1 1
300 400 500 600 700 800
PPM SULFUR
1
900
Figure 4.12. Mean values and 95% confidence intervals for sulfur in all plots. Upper crown, older trees,
both collection sets.
-------
1975 COLLECTION
1972
1975
1976 COLLECTION
h-
1974
1973
i
60
i
70
80
i
90
100
PERCENT NEEDLE RETENTION
Figure 4.13.
Mean values and 95% confidence intervals for percent needle retention, 1975 and 1976
collection, upper crown, for plot E-l.
-------
TABLE 4.22. BASIC STATISTICS FOR SULFUR CONCENTRATIONS IN UPPER CROWN
YOUNGER TREES AND UPPER CROWN OLDER TREES, ALL YEARS FOLIAGE
COMBINED, PLOT S-5*
Statistic
N
Maximum Value
Minimum Value
Mean
Variance
Standard Deviation
Standard Error
Upper Crown, Younger
15
1050
550
746
23023
151.7
39.1
95% Confidence Interval 84.0
Median
Standard Error
700
49. 1
Coefficient of Variation 20.3
Standard Error
3.8
Sampling Precision** 11.2
Upper Crown, Older
15
700
450
613
4809
69.3
17.9
38.4
650
22.4
11.3
2.0
6.2
*Di fference between mean values is significant @
between the individual years foliage were not si
pooled.
**Defined as (t „
X
5Sx') 100
p ^ .05. Since differences
gnificant, these values were
114
-------
TABLE 4.23. MEAN VALUES FOR TOTAL SULFUR FROM UPPER CROWN YOUNGER AND OLDER
TREES, ALL PLOTS, 1976 COLLECTION.
Plot & Mean
Sulfur
Concentration
Plot &
Mean Sulfur
Concentration
Younger Trees
S-3
SE-2 E-
S-5
SE-4
640
410
620
410
528
410
746
*.,
410
640
620
528
S-5
746
746
746
640
620
E-l
528
528
640
SE-2
620
Older Trees
Plot & Mean
Sulfur
Concentration
Plot &
Mean Sulfur
Concentration
S-3
SE-2 E-l
S-5
SE-4
590
388
603
388
533
388
613
388
603
S-5
533
613
613
603
E-l
533
590
SE-2
603
'Indicates mean values shown are significantly different @ p ^ .05
115
-------
necrosis for upper crown positions for the 1975 and 1976 collections are shown
in Figure 4.14. Although the mean values for 1974 year's foliage increased
slightly in 1976 over 1975, the values for 1973 foliage have not (Figure 4.14).
For percent mottled needles, the ANOVA for 1975 showed significant treat-
ment effects between the tree ages, while the ANOVAs for 1975-1976 and 1976
indicated significant treatment effects between the years of foliage, but the
ANOVA for 1975 did not.
The mean values and 95% confidence intervals for percent mottled needles
for the 1976 collections from plot S-3 are shown in Figure 4.15, which illus-
trates the lack of effect between the tree ages (compare Figure 4.9).
For percent total needle necrosis, the ANOVA for 1975-1976 detected
significant treatment effects between the two different years' collections and
between the years of foliage. The ANOVA for 1975 detected significant effects
only between the tree ages. The ANOVA for 1976 did not indicate differences
between the mean values for tree ages but did detect differences between the
years of foliage. For the individual years of foliage, the 95% confidence
intervals enclosed the mean values for the tree ages at plot E-l (Figure 4.16)
for the 1975 collection as well as the 1976 collection. The mean values and 95%
confidence intervals for older and younger trees combined are shown in Figure
4.16 for both collections from plot E-l. The 1976 data in Figure 4.16
illustrate the gradual increase in percent total necrosis fo the other plots in
1976.
For percent healthy needles, the ANOVA for 1975-1976 detected significant
treatment effects between the collections. The 1975-1976 ANOVA did not indi-
cate differences between the plots, nor did it for the 1975 collections. The
1976 ANOVA detected significant treatment effects between the plots. All
ANOVAs indicated significant differences between the years of foliage (Table
4.21). The mean values and 95% confidence intervals for percent healthy
needles for all plots from the 1976 collection are shown in Figure 4.17.
The results of the analysis of the data from the 1975 collections demon-
strated that samples collected from the upper or lower crown positions of
individual trees to evaluate the pre-operational baseline conditions for the 12
characteristics measured herein may be considered samples which are repre-
sentative of the individual tree as a whole, with the exception of fascicular
cross-sectional area. However, if such samples were collected from geograph-
ically separate plots, from different tree ages within a plot, or from differ-
ent ages of foliage, they cannot a priori be considered to be samples from the
same population.
Furthermore, the results of the data analysis for the 1976 collection and
the comparison of both years' collections show that, with the exception of
fluoride, percent total needle necrosis, and percent healthy needles, the two
collections represent samples from the same population, even though differ-
ences were indicated for certain variables within the two collections. For
example, treatment effects between the plots for sulfur concentrations were
noted in 1976 but not in 1975, and treatment effects between the years of
foliage were noted in 1976 but not in 1975. These effects, however, were not
116
-------
1974
1973
1975 COLLECTION
-\ 1972
(
j — •» 1 iy/o
1 , -A, i Q7/i i O7f, rni i rrTTnN
• „„,.,...„ 1 O7T
— — j iy/j
i i 1
3 5 10 15
1
20
PERCENT BASAL NECROSIS
Figure 4.14.
Mean values and 95% confidence intervals for percent basal necrosis, upper crown positions.
1975 and 1976 collections, plot E-l.
-------
oo
1 t
I f
1 *
1 *
1 t
1 '
1 -•
1 *
III III
0 5 10 15 20 25 30
YOUNGER TREES
1 Q73
| 1Q71 OLDER TREES
1 1 1 1 i
35 40 45 50 55
PERCENT MOTTLED NEEDLES
Figure 4.15.
Mean values and 95% confidence intervals for percent mottled needles for upper crown,
younger and older trees, plot S-3.
-------
I » I 1974
I > | 1973
1 '1975 COLLECTION
| • | 1972
^ 1975
PERCENT TOTAL NEEDLE NECROSIS
.j 1974 1976
COLLECTION
•I 1973
I i i i
8 10 12 14
Figure 4.16. Mean values for percent total needle necrosis for upper crown, older and younger trees
combined, 1975 and 1976 collections.
-------
PLOT
SE-4
1975
1974
PLOT
S-5
PLOT
E-l
1973
1975
^ 1974
^ 1973
1975
-j 1974
1973
PLOT
SE-2
1975
1974
1973
PLOT
S-3
1975
1974
^ 1973
20
40 60 80
PERCENT HEALTHY NEEDLES
I
100
Figure 4.17.
Mean values and 95% confidence intervals for percent healthy
needles, 1976 collection.
120
-------
sufficient to permit the conclusion that the two years' collections were
samples from different populations.
CHEMICAL ANALYSIS OF UNDERSTORY SPECIES
Table 4.24 depicts the results of fluoride and sulfur analyses and basic
statistics for the foliage of nine understory species collected at the five
permanent sites during both 1975 and 1976 collection periods. Also on this
table are the results of fluoride and sulfur analyses of ten other understory
species which were not abundant at all the ponderosa pine-skunkbush sites;
these species were collected during either 1975 or 1976 at one or more of the
five sites. The basic statistics for sulfur and fluoride content of these 19
total species is presented in Table 4.25. During 1977, eight separate samples
of each understory species from each site will be collected for chemical
analysis.
The mean fluoride concentration found for all understory species was below
5 ppm. Little bluestem and bluebunch wheatgrass, the two most common grass
species at the ponderosa sites, usually had the lowest fluoride levels (i.e. , <
2 ppm) of any understory species collected. Sulfur levels found in foliage of
silver sage, broom snakeweed, and arrowleaf balsamroot usually were higher than
other common understory species and two to three times higher than the two
commonly found grass species.
In a study of fluoride- and sulfur-polluted areas in which ponderosa pine
foliage and understory species of skunkbush, broom snakeweed, and bluebunch
wheatgrass were utilized, Gordon et aj_. (1977) demonstrated that the foliage of
the dominant species (ponderosa pine) accumulates excessive levels of both
sulfur and fluoride at a faster rate than the understory species during an
exposure time of 15 months. The researchers do not anticipate any significant
increase of fluoride or sulfur accumulation in understory species at the five
ponderosa pine-skunkbush sites until the ponderosa pine foliage reaches two to
three times the baseline for fluoride and 1.5 times the baseline for sulfur.
DISCUSSION AND SUMMARY
SULFUR
Several past studies of air pollution impacts on conifer species (i.e.,
Linzon, 1973a; Dreisinger, 1965; Ellertsen et aj_. , 1972; Berry et aJL , 1964;
Miller et aj_. , 1963; Carlson, 1972) have encompassed one to four of the
growth/health/disease characteristics (i.e., tip burn, needle retention,
needle length, and percent necrosis) which have been used in the Colstrip
ponderosa pine sites during the last two years to quantify the variability of
these characteristics prior to the incidence of air pollution damage. Also,
there have been several air pollution studies of conifer trees to determine
sulfur and/or fluoride concentrations in damaged and undamaged foliage (i.e.,
Katz et al_. , 1952; Carlson, 1974; Compton et al_. , 1961 ; Ellertsen et al_. , 1972;
Knabe, 1968; Linzon, 1972, 1973b).
Linzon's sulfur studies in the Sudbury area of Ontario revealed that 15-
month-old jack pine foliage (Pinus banksiana), growing 27 km northeast and 160
121
-------
TABLE 4.24. FLUORIDE AND SULFUR CONCENTRATIONS IN UNDERSTORY SPECIES FROM THE 1975 AND 1976 COLLECTIONS.
SE-4
Species
Silver Sage
Broom Snakeweed
Chokecherry
Skunkbush
Idaho Fescue
Bluebunch Wheatgrass
Prairie Sage
Arrowleaf Balsamroot
Little Bluestem
Lupine
ppm F
1975
1976
2.3
2.3
—
1.7
1.8
2.4
6.4
0.9
0.8
1.4
1.1
1.7
2.2
3.1
—
4.0
—
—
—
2.1
ppm S
1975
1976
1300
1950
950
650
800
950
900
700
950
800
950
900
1000
2400
1400
s-
ppm F
1975
1976
3.
3.
1.
--
--
--
1.
1.
--
2.
2.
0.
5.
4.
4.
4.
--
--
2.
2.
0
0
5
-
-
-
9
7
-
0
8
9
7
0
9
1
-
-
1
0
5
ppm S
1975
1976
1300
1550
1675
950
950
800
1100
750
1150
1000
1150
750
1100
950
E-l
ppm F ppm S
1975 1975
1976 1976
1.3 1300
1.8 1400
—
—
—
—
1.8 1800
700
—
—
1.1 500
_
0.9 1000
— _ —
—
—
2.5 300
3.3 250
—
5.5 1150
SE-2
ppm F
1975
1976
2.0
2.7
2.2
4.9
2.5
4.1
4.9
—
—
2.1
2.1
4.2
_ ~ _
—
—
2.3
1.2
—
2.1
ppm S
1975
1976
1400
1250
1800
2500
600
800
1000
600
600
1100
_ _ —
400
300
950
s-
ppm F
1975
1976
--
2.
--
2.
0.
4.
3.
2.
--
2.
--
2.
4.
— ~
--
—
1.
1.
4.
3.
-
5
-
3
7
7
6
0
-
2
-
5
0
~
-
-
2
0
9
4
3
ppm S
1975
1976
850
1200
300
500
600
600
550
700
650
"• — — ~
300
400
750
800
(continued)
-------
TABLE 4.24. (continued)
SE-4 S-5 E-1 SE-2 S-3
ppm F ppm S ppm F ppm S ppm F ppm S ppm F ppm S ppm F ppm S
Species 1975 1975 1975 1975 1975 1975 1975 1975 1975 1975
1976 1976 1976 1976 1976 1976 1976 1976 1976 1976
Scurf Pea --- --- 2.8 950 1.9 900 —
--- 4.7 1350 --- 2.9 900
Yucca --- 2.8 800 --- --- ---
1.9 700 5.6 750 — 1.6 650
Red Three-Awn --- 2.9 500 ---
--- 1.7 650 --- ---
Needle and Thread — — 2.7 700 2.2 1100 —
--- --- 4.4 1200 ---
Vetch --- --- 2.4 1200 — —
Fringed Sage — — — — —
2.6 1400 2.4 1250 5.3 1250 1.8 1100 1.9 1200
Green Rabbit Brush — — — — — —
2.0 1550 --- --- ---
Rocky Mountain Juniper — — — — —
— --- 600 1.9 800 —
Big Sagebrush — — — — —
--- —- 2.4 1300 ---
-------
TABLE 4.25. UNDERSTORY SPECIES, PONDEROSA PINE - SKUNKBUSH SITES.
Species and Year
of Collection
Silver Sage
Broom
Snakeweed
Chokecherry
Skunkbush
Bluebunch
Wheatgrass
Prairie Sage
Little
Bluestem
Lupine
Scurf Pea
Idaho
Fescue
Arrowleaf
Balsamroot
Yucca
Red Three-Awn
Needle-and-
Thread
Vetch
Fringed Sage
Common
Rabbi tbrush
Rocky Mountain
Juniper
Big Sage
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1975
1976
1976
1975
1976
Mean
F"
2.15
1.85
1.85
2.97
1.67
3.55
3.56
2.38
1.78
1.8
3.4
3.55
2.0
1.83
3.5
3.0
2.35
3.8
.8
1.87
4.9
4.05
2.8
3.03
2.9
1.7
2.45
4.4
2.4
2.8
2.0
1.9
2.4
S
1230
1400
1737
1550
516
650
1020
830
750
750
960
1000
333
316
925
1050
925
1125
700
766
1150
1575
800
700
500
650
900
1200
1200
780
1550
700
1300
Standard
Deviation
F~
.70
.49
.49
1.7
.91
1.63
1.88
1.75
.83
.68
1.87
.64
.7
1.27
1.97
1.5
.63
1.27
.42
.07
2.23
.35
1.44
S
216
403
88.3
832.1
189
212
459
171.7
264
147.2
198. 1
0
57.7
76.3
247
231.1
35.3
318
202
1166
50
—
—
282
713.6
141.4
Standard
Error
F"
.35
.35
.35
.98
.52
1.15
.84
.87
.42
.34
.84
.45
.40
.73
1.4
.67
.45
.9
.24
.05
1.29
.25
.64
S
96.7
180.2
62.5
480.4
109
150
205.3
76.8
132.2
73.6
88.6
0
33.3
44.0
175
103.6
25
225
116.6
—
825
—
28.8
—
—
200
—
—
318. 1
* —
100
—
124
-------
km west of the non-ferrous smelters, contained sulfur averaging 2,200 to 1,000
ppm, respectively. Jack pine foliage of the same age collected at two inter-
mediate-distance sampling sites, 45 and 67 km northeast, had average sulfur
concentrations of 1,700 and 1,300 ppm, respectively, while three-month-old
foliage from these same two intermediate sites had average sulfur concentrations
of 1,500 to 1,400 ppm respectively.
Katz ejt al. (1952) worked with both field-grown and potted ponderosa pines
and fumigated with S02 concentrations of 22 to 500 pphm for durations of 35 to
1,656 hours (1.4 to 69 possible day periods), respectively. These studies
revealed that the most elevated sulfur levels (5,600 ppm) were found in
one-year-old foliage exposed to the lowest concentration (22 pphm) for the
longest period of time (1,656 hours).
Katz's fumigation studies also revealed that one-, two-, and three-year-
old ponderosa pine foliage fumigated for 96 hours with S02 concentrations of 500
pphm contained 120, 40, and 300 ppm of sulfur, respectively. In these studies,
there appeared to be a relationship between excessive sulfur accumulation and
the incidence of the S02-caused needle necrosis that Katz was attempting to
determine. For instance, ponderosa pine trees fumigated with 500 pphm of sulfur
for 96 hours developed no foliage necrosis but had 530 ppm of sulfur in the one-
year-old needles and 520 ppm in the two-year-old needles after the fumigation.
In another S02 fumigation study utilizing 500 pphm for 40 hours, Katz found that
100 percent necrosis occurred, and in the one-, two-, and three-year-old
foliage, the sulfur levels were found to be elevated above 750 ppm.
In all of Katz's, as well as Linzon's (1958 to 1973) reported S02 field and
fumigation studies, no details were presented on the variation of sulfur
concentrations in healthy foliage of pines within and between the different
years' growth. Thus, it is difficult to compare the variability in sulfur
concentrations of ponderosa pine from the Col strip study area with either of
their studies.
Another field study in the area of the Trail, B.C., smelter (besides that
of Katz) was carried out by U.S. Forest Service personnel (Sheffer and Hedgcock
1955) between 1928 and 1936. One of the forest species collected and studied by
these two investigators was ponderosa pine. Sulfur analysis of damaged and
undamaged ponderosa pine foliage revealed that foliage with concentrations of
2,700 to 2,300 ppm collected at sites 18 and 37 km, respectively, south of the
emission source were damaged but that foliage with 2,000 to 1,600 ppm collected
42 to 70 km, respectively, south of the source were undamaged. Baseline sulfur
content of healthy ponderosa pine foliage collected by Scheffer and Hedgcock 79
to 97 km south of the source was found to range from 800 to 1,000 ppm. Over
25,000 ponderosa pine trees were utilized in this eight-year field study, and
the various types of damage include decreased cone production and seedling
reproduction, loss of annual increment, premature needle casting, and increased
sulfur content of the foliage. This study was one of the first and only field
studies in which the investigators set up their studies and carried them out to
compare and understand the variabilities of pathologies at control sites versus
the impact sites.
125
-------
Some field investigations during the last 15 years, such as Carlson's
(1974) study on damage to ponderosa pine from a small sulfur dioxide source in
the Missoula Valley of Montana, demonstrated that elevated sulfur content of
pine (Pinus ponderosa) is related to the severity of the foliage damage; other
studies, such as those by Ellertsen (1972) and Berry and Hepting (1964) on
sulfur and fluoride levels in damaged white pine (Pinus strobus) foliage in
Tennessee, did not demonstrate this.
To date, after an intensive search through the sulfur dioxide pollution
literature involving both field and controlled fumigation chamber investiga-
tions, the investigators have not found any studies dealing with S02 damage to
plants that quantify the variability of baseline sulfur content within foliage
of any conifer species between different years' foliage or between foliage from
different control sites. However, numerous field investigators and some
control fumigation investigators have utilized the sulfur content of S02-
damaged foliage as one of the major indices of their pollution studies. If the
variability of the sulfur content of foliage from controlled areas is not
known, then sulfur levels in damaged foliage is much less useful for under-
standing if and when airborne sulfur emissions are affecting ecosystems.
Every field investigator mentioned above used at least one foliage
pathology or characteristic to demonstrate air pollution impact on the study
sites. Also, each was studying an area which was being severely impacted by
the emissions from one or more large stationary sources. Investigations in
areas where acute fumigation damage is occurring may not require an under-
standing of the baseline quantities of needle pathologies or the variability of
those pathologies between different years' foliage, different-sized trees,
different crown positions, and different sites. However, these baseline
quantities are absolutely essential for understanding if and where chronic air
pollution damage is occurring within ecosystems.
The baseline studies on the ponderosa pine sites clearly demonstrated that
abiotic and biotic causal agents normally occurring in a pristine area cause a
measurable amount of damage to pine foliage; this damage has to be quantified
prior to understanding if and where chronic air pollution damage is occurring.
FLUORIDE
Fluoride studies from both field and fumigation chamber investigations
have revealed that this element is the most phytotoxic of the normal gaseous
emissions from large stationary sources of air pollution (Lione et al_. , 1962).
More controlled S02 fumigation studies have been completed and published than
fluoride studies, but there are more published field studies dealing with
fluoride pollution than with any other phytotoxic gas because: (1) Gaseous and
particulate fluorides are the major emissions of large stationary sources such
as aluminum and steel smelters, and phosphate fertilizer, animal supplement,
and P205 plants; (2) in most cases, baseline fluoride levels in vegetation are
below 10 ppm, and fluorine has not proven to be an essential ion for plant
metabolism or structure, and (3) fluoride pollution problems, in many past
instances, have involved domestic animals or agricultural crops, provoking
intensive scientific investigation.
126
-------
Fluoride is not a major pollutant within the emissions of coal-fired power
plants or non-ferrous smelters, and few studies around either pollution source
have included this element or compound. However, because of the phytotoxicity
of even low concentrations (.5 ppb) of fluoride in the ambient air to plant
pollen (Facteau et al., 1973, 1977), its low translocation rate from the leaf
receptor to other parts of the plant (thus remaining in the leaf), and its
rapid build-up in the food chain, the investigators have spent considerable
time determining the baseline levels of this element in the ponderosa pine-
skunkbush ecosystems of the Colstrip area.
The fluoride studies in upper crown foliage at the ponderosa sites were
set up to best utilize the evidence gained by Knabe (1968) in forest canopy
fluoride studies which demonstrated that the most elevated fluorides in conif-
erous foliage were in the highest and most exposed upper crown foliage. Field
studies in the vicinity of aluminum plants by various investigators (Shaw et
al_. , 1951; Adams et al_. , 1952; Compton et al_. , 1961; Facteau et al_. , 1976;
Carlson, 1972; Gordon, 1974) in the United States have demonstated that pon-
derosa pine is very susceptible to fluoride pollution. Current and one-year-
old foliage of ponderosa pine with fluoride levels less than 20 ppm have been
reported to manifest needle tip necrosis by Compton et al_. (1968), Carlson
(1972), Hindawi (1973), and Gordon et ah (1976).
Partitioned ponderosa pine needles damaged by fluoride pollution have
been reported by Gordon et al_. (1976) to contain up to three times more
fluoride in the 3 cm tip portion than whole needles and over six times more
fluoride in the needle tips than middle and basal portions. Both Jacobson et
aj_. (1966) and Compton et al_. (1960) have demonstrated with fluoride-fumigated
gladiolus plants that after fluoride enters the leaves, it is translocated to
the leaf tips where tissue necrosis then occurs. Jacobson1s (1966) fluoride
fumigation studies with other species of plants (cotton and tomato) to
determine fluoride translocation and partitioning in plant tissues led him to
suggest that the ability of a plant to readily translocate and partition
fluoride may influence its susceptibility or resistance to fluoride damage.
The 1976 data from this study demonstrated an increase in fluoride con-
centrations in ponderosa pine foliage from Kluver (E~l), McRae (SE-2), and Fort
Howes, Custer National Forest (S-5) sites over that occurring in foliage of the
same age during 1975. This was not accompanied by any significant increase in
sulfur levels in the needles at these three sites between the 1975 and 1976
study periods. Because sulfur taken in from the ambient air is translocated
out of the pine needles, no significant increase is expected at any of these
sites until after the emissions of the Colstrip coal-fired power plant units
increase from the estimated average 1976 level of .8 tons per hour to the
higher potential levels of +2 tons per hour.
NEEDLE PATHOLOGIES
While there was a significant increase in both the amount of total needle
necrosis and loss of healthy needles at, all the ponderosa pine-skunkbush sites
during 1976 over that of 1975, at this time the increase is not attributed to
phytotoxic emissions of Colstrip units. Houston (1974), fumigating white pine
with a combination of S02 (.025 ppm) and 03 (.05 ppm) for six hours, reported
127
-------
that all sensitive clones manifested tip necrosis on 75 to 100 percent of
current year needles. However, these are hypersensitive clones of white pine
and are much more sensitive to both of these gases than ponderosa pine (Miller
et al_. , 1975). Constantindou et al_. (1976) reported a significant decrease in
the chlorophyll content of red pine (Pinus resinosa) cotyledons when fumigating
with 0.5 ppm S02 for both 15-and 30-minute periods. Kress and Skelly (1977),
when fumigating loblolly pine (Pinus taeda) with S02 (0.14 ppm) + 03 (.05 ppm) +
N02 (0.10 ppm) for six hours per day for 28 consecutive days reported a 26
percent reduction in growth. The studies of both Constantindou et
-------
REFERENCES
Adams, D. F. , D. J. Mayhew, R. M. Gnagy, E. P. Richey, R. K. Koppe, and I. W.
Allan. 1952. Atmospheric pollution in the ponderosa pine blight area,
Spokane County, Washington. Ind. Eng. Chem. 44:1356-1365.
Berry, C. R. and G. H. Hepting. 1964. Injury to eastern white pine by
unidentified atmospheric constituents. Forest Sci. 10:1-13.
Carlson, C. E. 1972. Monitoring Fluoride Pollution in Flathead National Forest
and Glacier National Park. U.S. Forest Service, Division of State and
Private Forestry, Missoula, Montana, pp. 1-25.
Carlson, C. E. 1974. Sulfur Damage to Douglas-Fir Near a Pulp and Paper Mill in
Western Montana. U.S. Forest Service, Division of State and Private
Forestry, Missoula, Montana. Report No. 74-13. 41 pp.
Compton, 0. C. and L. F. Remmert. 1960. Effects of air-borne fluorine on injury
and fluorine content of gladiolus leaves. Proc. Amer. Soc. Hort. Sci.
75:663-675.
Compton, 0. C., L. F. Remmert, and J. A. Rudinsky. 1961. Needle Scorch and
Condition of Ponderosa Pine Trees in The Dalles Area. Ore. Agr. Exp. Sta. ,
Oregon State University, Corvallis. Misc. Paper 120.
Compton, 0. C., F. W. Adams, W. M. Mellenthin, S. Elliott, N. Chestnut, and D. W.
Bonney. 1968. Fluorine Levels in Crops of The Dalles Area, 1965-67:
Cherry, Peach, and Pine Trees and the Ambient Air. Ore. Agr. Exp. Sta.,
Oregon State University, Corvallis. Spec. Rpt. 261. 40 pp.
Constantindou, H. , T. T. Kozlowski, and K. Jensen. 1976. Effects of sulfur
dioxide on Pinus resinosa seedlings in the cotyledon stage. J. Env.
Quality. 5:141-144.
Dreisinger, B. R. 1965. Sulfur Dioxide Levels and Effects of the Gas on
Vegetation Near Sudbury, Ontario. Presented at 58th Annual Meeting of the
Air Pollution Control Association. Paper No. 65-121.
Ellertsen, B. W. , C. J. Powell, and C. L. Massey. 1972. Report on study of
diseased white pine in Tennessee. Mitt. Forstl. Bundesversuchanst., Wien.
97:195-206.
Environmental Protection Agency. 1976. The Bioenvironmental Impact of a Coal-
Fired Power Plant, Second Interim Report, Colstrip, Montana—June 1975 (R.
A. Lewis, N. R. Glass, and A. S. Lefohn, eds.). Environmental Research
Laboratory, Office of Research and Development, Corvallis, Oregon. EPA-
600/3-76-013.
Facteau, T. J. , S. Y. Wang, and K. E. Rowe. 1973. The effect of hydrogen
fluoride on pollen germination and pollen tube growth in Prunus avium L.
cv. "Royal Ann." J. Amer. Soc. Hort. Sci. 98(3): 234-236.
129
-------
Facteau, T. J. and W. M. Mellenthin. 1976. Fluoride Investigations in The
Dalles Area 1968-1974. Ore. Agr. Exp. Sta., Oregon State University,
Corvallis. Tech. Bull. 132. 56pp.
Facteau, T. J. and R. E. Rowe. 1977. Effect of hydrogen fluoride and hydrogen
chloride on pollen tube growth and sodium fluoride on pollen germination in
"Tilton" apricot. J. Amer. Soc. Hort. Sci. 102(1):95-96.
Gordon, C. C. 1974. Environmental Effects of Fluoride: Glacier National Park
and Vicinity. U.S. Environmental Protection Agency, Region VIII, Denver,
Colorado. EPA-908-1-74-001. 150pp.
Gordon, C. C. , P. C. Tourangeau, and C. E. Carlson. 1976. Fluoride Emissions of
Coal-Fired Power Plants and their Accumulation in and/or Impact Upon Plant
and Animal Species. Paper presented at American Chemical Society
Symposium, San Francisco, California. 10 pp.
Gordon, C. C., P. C. Tourangeau, J. J. Bromenshenk, C. E. Carlson, and P. M.
Rice. 1977. Pre- and Post-Operational Investigations into the Impacts of
Coal-Fired Power Plant Emissions in the Northern Great Plains. Paper
presented to the National Academy of Sciences/National Research Council
Meeting, Washington, D.C. 85 pp.
Hindawi, I. J. 1973. Fluoride in Glacier National Park: A Field Investiga-
tion. U.S. Environmental Protection Agency, Region VIII, Denver,
Colorado. EPA-908-1-73-001. 69pp.
Houston, D. B. 1974. Response of selected Pinus strobus L clones to
fumigations with sulfur dioxide and ozone. Can. J. For. Res. 4:65-68.
Jacobson, J. S. , L. H. Weinstein, D. C. McCune, and A. E. Hitchcock. 1966. The
accumulation of fluoride by plants. J. Air Poll. Contr. Assoc. 16:412-
417.
Jones, B. L. and C. C. Gordon. 1965. Embryology and development of the
endosperm haustorium of Arceuthobium Douglasii. Amer. J. Bot. 52(2): 127-
132.
Katz, M. and A. W. McCallum. 1952. The effects of sulfur dioxide an conifers.
Proc. U.S. Technical Conference on Air Pollution (New York, Toronto,
London), pp. 84-96.
Kay, C. E., P. C. Tourangeau, and C. C. Gordon. 1975. Fluoride levels in
indigenous animals and plants collected from uncontaminated ecosystems.
Fluoride. 8(3):125-131.
Knabe, W. 1968. Experimentelle Prufung der Fluoranreicherung in Nadeln und
Slattern von Pflanzen in Abhangigkeit von deren Expositionshoh uber Grund.
Miedzynarodowej Konf., Wplyw. Zanieczyszczen Powietrza N Lasy, 6th,
Katowice, pp. 101-116.
13Q
-------
Kress, L. W. and J. M. Skelly. 1977. The Interaction of 03, S02 and N02 and its
Effects on the Growth of Two Forest Tree Species. Cotterell Centennial
Symposium—Air Pollution and Its Impact on Agriculture, pp. 128-152.
Leone, I. A., E. Brennan, and R. H. Daines. 1962. Effects of Air Pollution on
Vegetation. Presented at the Air Pollution Control Association, Mid-
Atlantic States Section Meeting, Wilmington, Delaware. 6 pp.
Linzon, S. N. 1958. The Influence of Smelter Fumes on the Growth of White Pine
in the Sudbury Region. Joint Pub., Ont. Dept. Land Forests, Ontario Dept.
Mines, Toronto, Ontario. 45 pp.
Linzon, S. N. 1972. Pre-Pol1ution Background Studies in Ontario. Proc. 27th
Ann. Meeting of the Soil Conservation Society of America, pp. 1-7.
Linzon, S. N. 1973a. The Effects of Air Pollution of Forests. Paper presented
at the 4th Jt. Chem. Eng. Conf. pp. 1-18.
Linzon, S. N. 1973b. Sulfur Dioxide Air Quality Standards for Vegetation.
Paper presented at the 66th Annual Meeting of the Air Pollution Control
Association. Paper No. 73-107. 19 pp.
Materna, J. 1973. Criteria for Characterization of Pollution Damages in
Forests. Proc. Int. Clean Air Congr. 3rd, Dusseldorf, West Germany, pp.
A121-A123.
Maughan, D. 1977. Personal Communication and monthly computer readouts for
S02, 03, N02, HF, and particulate for the Burlington Northern air
monitoring site at Colstrip, Montana.
Miller, P. R. , J. R. Parmeter, Jr., 0. C. Taylor, and E. A. Cardiff. 1963.
Ozone injury to the foliage of ponderosa pine. Phytopathology. 53:1072-
1076.
Miller, P. R. and J. R. McBride. 1975. Effects of air pollutants on forests.
Chapter 10, pp. 195-235. j.n Responses of Plants to Air Pollution (J. B.
Mudd and T. T. Kozlowski, eds.). Academic Press, New York.
Montana State Department of Health and Environmental Sciences. 1975. Findings
of Facts and Conclusions of Law. Colstrip Units 3 and 4 Proceedings Before
the Board of Natural Resources and Conservation, and the Board of Health
and Environmental Sciences. 3 pp.
Northern Cheyenne Research Project. 1976. The Northern Cheyenne Air Quality
Redesignation Report and Request. Prepared for the Environmental
Protection Agency, Region VIII, Denver, Colorado, pp. 5-6.
Scheffer, T. C. and G. G. Hedgcock. 1955. Injury to Northwestern Forest Trees
by Sulfur Dioxide from Smelters. U.S. Forest Service Tech. Bull. 1117:1-
49.
131
-------
Shaw, C. G. , G. W. Fischer, D. R. Adams, M. F. Adams, and D. W. Lynch. 1951.
Fluoride injury to ponderosa pine: A summary. Northwest Sci. 15:156.
Sokal, R. S. and F. J. Rohlf. 1969. Biometery, the Principles and Practices of
Statistics in Biological Research. W. H. Freeman and Company, San
Francisco. 775 pp.
132
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APPENDIX A
LESS THAN ONE-HOUR EXPOSURE OF SO? ALONE AND IN COMBINATION WITH OTHER POLLUTANTS
TIME
CONCENTRATION RESPONSE OF VEGETATION
PLANT SPECIES
AUTHOR
1. 15 min. .5 ppm S02
CO
CO
Significant decrease in chlorophyll content of
cotyledons. Significant decrease in chloro-
phyll content of cotyledons at 1, 3, and 4 ppm.
Highly significant reductions in chlorophyll
in primary needles at .5, 1, 3, and 4 ppm.
Highly significant decrease in increment of
total dry weight (growth)of cotyledons at .5,
1, 3, and 4 ppm. Highly significant decrease
in increment of total dry weight (growth) of
primary needles at .5, 1, and 3 ppm; signifi-
cant decrease reduction at 4 ppm.
No necrosis of tips of cotyledons and primary
needles at 1 day after treatment of .5, 1, 3,
or 4 ppm.
Red pine
seedlings
Pinus resinosa
Constant!ndou
et al., 1976
2. 30 min. .5 ppm S02
Significant decrease in chlorophyll content of
cotyledons. Highly significant decrease in
chlorophyll content of cotyledons at 1, 3, and
4 ppm.
Highly significant decrease in chlorophyll
content of primary needles at .5, 1, 3, and
4 ppm.
High-ly significant decrease in increment of
total dry weight (growth) of cotyledons at
.5, 1, 3, and 4 ppm. Highly significant
decrease in increment of total dry weight
(growth) of primary needles at .5 and 1 ppm;
significant at 3 and 4 ppm.
Red pine
seedlings
Pinus resinosa
Constantindou
et al., 1976
-------
APPENDIX (continued)
TIME
CONCENTRATION RESPONSE OF VEGETATION
PLANT SPECIES
AUTHOR
37% of seedlings had necrosis of tips of
cotyledons and primary needles 1 day after
exposure to 4 ppm.
3. 1 hour .5 ppm S02 Significant decrease in chlorophyll content
of
in
3,
cotyledons.
chlorophyl 1
and 4 ppm.
Highly
content
significant decrease
of cotyledons at 1 ,
Red pine Constant! ndou
seedlings et al_. , 1976
Pinus resinosa
00
-Pi
Highly significant decreases in chlorophyll
of primary needles at .5 and 1 ppm; signi-
ficant decreases at 3 and 4 ppm.
Highly significant decrease in increment of
total dry weight (growth) of cotyledons at
3 and 4 ppm. Exposures .5 and 1 ppm were
significant. Highly significant decreases
in increment of total dry weight (growth) of
primary needles at .5 and 1 ppm; significant
at 3 and 4 ppm.
15% of seedlings had necrosis of tips of
cotyledons and primary needles 1 day after
exposure to 3 ppm; 25% had necrosis after
exposure to 4 ppm. "Continuous fumigation
with S02 at much lower dosages than used
may have inhibitory effects on seedling
development and on regeneration of plant
communities."
-------
APPENDIX (continued)
TIME
CONCENTRATION RESPONSE OF VEGETATION
PLANT SPECIES
AUTHOR
CO
en
15 min. .5 ppm S02 Highly significant decrease in chlorophyll
to 2 content of cotyledons for .5, 1, 3, and 4
hours. ppm.
Highly significant decreases in chlorophyll
content in primary needles.
Highly significant decrease in increment of
total dry weight at 3 and 4 ppm. Exposures
to .5 and 1 ppm were significant.
Highly significant decrease in increment of
total dry weight (growth) of primary needles
at .5 and 1 ppm; significant at 3 and 4 ppm.
15% of seedlings had necrosis of tips of
cotyledons and primary needles 1-day after
exposures to 3 ppm; 35% after exposure to
4 ppm.
Red pine
seedlings
Pinus resinosa
Constant!ndou
et al., 1976
5.
6.
7.
2
2
2
hours
hours
hours
0.
0.
0.
25
25
05
ppm
ppm
ppm
S02
S02
S02
6.5% of exposed parts i
lesions.
4.5% of exposed parts i
lesions.
Caused tip necrosis on
njured. Needle
njured. Needle
new needles.
Eastern white
pine
Pinus strobus
Jack pine
Pinus banskiana
East. White pine
Pinus strobus
Berry,
Berry,
Costoni
1973
1971
1971
s,
-------
APPENDIX (continued)
TIME
CONCENTRATION RESPONSE OF VEGETATION
PLANT SPECIES
AUTHOR
2 hours
Alone
0.04 ppm
Followed 3 min
later by 0.05
ppm S02 for 2
hrs. and then
fumigated 24
hrs. later
with a mix-
ture at same
concentration.
[Needle necrosis (author stated necrosis
was most severe at this exposure).]
co
8. 4 hours
9. 6 hours
per day
for 28
days
0.05 ppm S02
+ 0.05 ppm
ozone
0.14 ppm S02
+ 0.05 ppm
ozone +
0.10 ppm N02
Severe needle necrosis (2-3 cm leaf tip
injured) on new needles, expressed most
vividly 72 hours after exposure.
OH TM PHMDTMATTnM \JTTU MTUCD DHI 1 IITAMTC
01/2 IN LUMB1NA 1 1UN Wll n UmhK PULLU 1 AN 1 o
Significant growth reduction (measured as
height) compared to either ozone and sulfur
dioxide combined, or ozone alone. Needles
were significantly narrower than for any
other exposure. This study is an example
of growth reduction with slight foliar
symptoms. Foliar symptoms response most
sensitive in early July.
East. White pine
Pinus strobus
Loblolly pine
Pinus taeda
(2 wks old)
Sycamore
(1 wk old)
Seedl ings
Costonis ,
1973
Kress and
Skelly,
1977
-------
APPENDIX (continued)
TIME
CONCENTRATION RESPONSE OF VEGETATION
PLANT SPECIES
AUTHOR
10. .6 hours 0.025 ppm
S02 + 0.05
ppm ozone
20-28 day-old white pine needles on ramets,
exposed 9 am - 3 pm. All sensitive clones
adversely affected. Response judged in terms
of needle elongation (growth) and foliar
lesion or tip necrosis. Author calls
response synergistic. 0.025 ppm S02 or 0.05
ppm = threshold; 0.10 ppm 03 = 20% necrosis.
East. White pine
Pinus strobus
Houston,
1974
11. 22 hours. 100 to 500
ppm S02
aqueous
sol.
Swelling of thylakoid disks in chloroplasts
disintegration of intro-chloroplast membranes
in mature needles. Absence of mitochondria.
Author state 100 - 500 ppm aqueous S02 is
equivalent to 1000-fold lower concentration
in air or 0.1 - 0.5 ppm.
Lodgepole pine
Pinus contorta
Malhotra,
1976
CO
Older needles more sensitive. Photosynthesis
reduced/measured by evolution of 03.
12. Annual 0.06
Average ppm
of 24-
hour
measurements
- 0.009 20 ± 5% loss in radial growth. Premature
drop of some needles. Reduction of above-
ground plant organs that can lead to impor-
tant losses of water in catchment areas (at
Hamr air monitoring station, Erzgebirge,
Germany). At higher S02 levels (0.026 - 0.037
ppm, annual average) rapid death of individual
trees and whole groups of trees (at Brandov
air monitoring station, Erzgebirge, Germany).
Fir forests
Abies sp.
Materna,
1973
-------
APPENDIX (continued)
GO
00
TIME CONCENTRATION
13. Average 0.045 ppm
30 min.
S02 cone.
for period
of 1954-
1963
14. 4-8 hr/ 0.1 ppm
day, 5 S02 + 0. 1
days/wk ppm ozone
4-8 weeks
RESPONSE OF VEGETATION
Foliar injury, premature defoliation,
reduced photosynthesis, decreased radial
and volume growth - 10 cubic feet per acre
per year ($1,171,000 in 1963), premature
death, rough bark, purple bark, reduced
blister rust, small leaf growth, more severe
symptoms with high pollution levels. 750
square miles in Sudbury, Ontario.
16% needle necrosis (chlorotic, yellow
spots, current year needles thin and twisted)
shedding of older needles far exceeded
damage responses from single exposures.
.1 ppm S02 injured 4% of needle area; .1 ppm
03 injured 3% of area.
PLANT SPECIES AUTHOR
East. White pine Linzon,
1971
Pinus strobus
East. White pine Dochinger
et al . ,
Pinus strobus 1970
-------
REFERENCES - APPENDIX
Berry, Charles R. 1971, Relative Sensitivity of Red, Jack and White Pine
Seedlings to Ozone and Sulfur Dioxide. Phytopathology. Vol. 61. pp. 231-
232.
Berry, Charles R. 1971. Age of Pine Seedlings with Primary Needles Affects
Sensitivity to Ozone and Sulfur Dioxide. Phytopathology. Vol. 64. pp.
207-209.
Constantindou, H.,T. T. Kozlowski, and K. Jensen. 1976. Effects of Sulfur
Dioxide on Pinus resinosa Seedlings in the Cotyledon Stage. Journal of
Environmental Quality. Vol. 5. pp. 141-144.
Costonis, Arth . 1971. Effects of Ambient Sulfur Dioxide and Ozone on
Eastern Wl K>- Pine in a Rural Environment. Phytopathology. Vol. 6. pp.
717-720.
Costonis, Arthur C 1973, Injury to Eastern White Pine by Sulfur Dioxide and
Ozone Alone ^ .; in Mixtures. European Journal of Forest Pathology. Vol.
3. pp. 50-55.
Dochinger, L. S. F w Rpnder, F. L. Fox, and W. W. Heck. 1970. Chlorotic Dwarf
on Eastern White s'ine caused by an Ozone and Sulfur Dioxide Interaction.
Nature. Vol. 225. .January 31. p. 476.
Houston, Daniel B. 19/4. Response of Selected Pinus strobus L. Clones to
Fumigations with Sulfur Dioxide and Ozone. Canadian Journal of Forest
Research. Vol. 4. pp. 65-68.
Kress, Lance W. and John M Skelly. January 13, 14, 1977. The Interaction of
03, S02, and N02 and Its Effect on the Growth of Two Forest Tree Species.
Cottrell Centennial Symposium, Air Pollution and Its Impact on
Agriculture, pp. 81-86.
Linzon, Samuel N. 1971. Economic Effects of Sulfur Dioxide on Forest Growth.
Journal of the Air Pollution Control Association. Vol. 21: No. 2. pp. 81-
86.
Malhotra, S. S. 1976. Effects of Sulfur Dioxide on Biochemical Activity and
Ultrastructural Organization of Pine Needle Chloroplasts. New
Phytologist. Vol. 76. pp. 239-245 plus plates.
Malhotra, S. S. and D. Hocking. 1976. Biochemical and Cytological Effects of
Sulfur Dioxide on Plant Metabolism. New Phytologist, Vol. 76. pp. 227-
237.
Materna, Jan. 1973. Criteria for Characterization of Pollution Damage on
Forests. Proc. Int. Clean Air Conqr. 3rd; Dusseldorf, W. Germany, pp.
A121-A123.
139
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SECTION 5
INVESTIGATION OF THE IMPACT OF COAL-FIRED POWER
PLANT EMISSIONS UPON INSECTS: ENTOMOLOGICAL
STUDIES IN THE VICINITY OF COLSTRIP, MONTANA
by
J. J. Bromenshenk
INTRODUCTION
Although this report concentrates on the entomological investigations in
the vicinity of Colstrip, Montana, from June 1, 1975, to March 1, 1977, it also
includes a synopsis of earlier reports and, therefore, serves as the major
report of the baseline phase (pre-operational) and monitoring phase (post-
operational) of the study. The rationale, methodologies, and preliminary
results were published in the Second Interim Report (EPA, 1976). Insect studies
at the sulfur dioxide treatment plots (ZAPS) are presented in section 14 of this
publication, while related fungal and vegetational investigations are
elaborated in sections 4 and 13.
The unifying hypothesis of the EPA-funded Coal-Fired Power Plant Project is
that methods can be developed to predict the bioenvironmental impacts of coal-
fired power plants before any damage occurs. The basic assumption is that air
pollutants are major contributors to damage, defined as negative changes in the
components and processes of an ecosystem. Our ancillary hypothesis is that
methods can be developed to serve this purpose, based on the use of indicator
species of plants, fungi, and insects as early warning systems and as continuous
monitors of atmospheric pollutants.
The southeastern Montana area was essentially free of anthropogenic
pollutants in the ambient air prior to the construction of the Colstrip power
plants (U. S. Environmental Protection Agency (EPA), 1976; Montana Department of
Health and Environmental Sciences (DHES), 1972; Montana Department of Natural
Resources and Conservation (DNRC), 1974; Westinghouse Environmental Systems
(WES), 1973). Therefore, it was hypothesized that if we measured, quantified,
and mathematically described pollutant-sensitive characteristics of key
indigenous species of the flora and fauna of the major ecosystems (cool season-
short grass and ponderosa pine-skunkbush) prior to the operation of the Colstrip
generating units, we could monitor any observable changes in these
characteristics as the ecosystems were subjected to pollutant challenge. Also,
by the appropriate analysis of temporal and spatial factors, we hoped to
correlate these changes with the level of air pollution and relate them to other
ecosystem components.
140
-------
The uniqueness of this approach lies in the opportunity it presents to
participate in a multi-disciplinary, multi-agency study, to conduct baseline
studies in a basically uncontaminated area, and to pursue these studies during
initial and subsequent impact of emissions from the Colstrip power plants. The
use of biological indicators or estimators is not new, and the rationale for
their use was stated well by Thomas e_t al. in a 1974 review of biological
indicators of environmental quality:
A continuous monitor of any environmental variable is superior
to periodic sampling because the concentration of pollutants or
intensity of stress varies with time. Because organisms integrate
their responses through time and because they react to all synergistic
and antagonistic effects of combined pollutants or stresses, they do
provide convenient full-time monitors. These biological indicators
give the actual responses of individual organisms or populations,
rather than predict biological responses from physical measurements
obtained through instrumentation.
However, it is essential that these indicators be sensitive to pertur-
bations, practical to monitor and measure, and reliable predictors of ecological
and economic impacts. To satisfy these criteria and to test the previously
stated hypothesis, we proposed and carried out, in progressive order, these
study objectives:
(1) Selection and establishment of permanent study sites (ponderosa pine-
skunkbush, grassland, and apiary) distributed to maximize gradient exposures to
the Colstrip emissions;
(2) compilation of inventories of insect and fungal populations, infes-
tations, and damage to indigenous plant species at the study sites to provide a
basis for the selection of more specific groups for use as biological
estimators;
(3) analysis and selection of insect and fungal populations with a
diversified but understandable interrelationship with indigenous plant species
in order to examine effects of air contaminants on symbiotic associations;
(4) selection and testing of disease- and injury-causing fungal and insect
species to be utilized for more intensive study;
(5) selection and testing of beneficial fungal and insect species to be
utilized for more intensive study;
(6) chemical analysis of the selected indigenous plants, insects, and fungi
to establish baseline levels and to discern any accumulation of materials
(fluorides and sulfurs) released by the conversion of coal to electricity, and
(7) determinations of physical (pH) and chemical characteristics of
precipitation.
After the inventory, selection, and testing phase, baseline data during
1974 and 1975 was gathered to determine:
141
-------
(1) Growth, health, and pathologies of ponderosa pines, including
associated fungal and insect populations and damage parameters;
(2) concentrations of fluorides and sulfurs in the dominant species of
indigenous plants of both the ponderosa pine and the grassland ecosystems;
(3) concentrations of these substances in an insect pollinator (honeybees)
of grasslands and croplands, and
(4) precipitation chemistry.
In 1976 the precipitation studies were abandoned, the number of pine study
plots were reduced and studies were intensified at the S02 fumigation (ZAPS)
plots. The objective was to increase the depth, understanding, and precision of
hypotheses regarding those systems, processes, and species which preliminary
studies indicated were the components most sensitive or vulnerable to measurable
pol1utant stress.
INSECTS
At least 1,400 species of insects inhabit the grassland ecosystems of
Montana (personal communication, N. Reese, USDA Rangeland Research Laboratory,
Bozeman, Montana). From among these, we elected to study: (1) Pest and
beneficial insects associated with ponderosa pine, the dominant tree species in
the Fort Union Basin, and a species susceptible to damage by phytotoxic gases
and acid rains (EPA, 1973); (2) pollinators, especially honeybees, and (3)
ground-dwelling beetles, particularly members of the families Carabidae,
Silphidae, and Scarabaeidae (see section on ZAPS). These three sets were chosen
because of probable sensitivity to air pollution and ecological and economic
importance.
A brief review of the economic considerations follows. Timber resources,
composed entirely of one commercial species, ponderosa pine, are harvested from
the Custer National Forest and the Northern Cheyenne Reservation. Based on
allowable annual cut and timber sale data, these pine stands have a wholesale
value of $1,584,000 ($180 per 1,000 board feet) (Gordon, 1975). Harmful insects
attack healthy or weakened trees, impairing vitality and causing mortality.
Bark beetles, historically, have destroyed more standing timber in western
forests than all other insects combined, and defoliators have ranked second as
destructive agents (Keen, 1952).
Agriculture is Montana's leading industry, according to annual reports of
the Montana Department of Agriculture Statistical Service. In 1973 the cash
receipts for the seven-county area of southeastern Montana totaled $110,042,600
or 10 percent of Montana's marketable produce. Pollinating insects are vital to
various crops and indigenous plant species. The honeybee is important not only
to Montana's agriculture, but also to that of California and other states where
25 percent of Montana's bees are taken in the winter for pollination purposes.
Montana is an important contributor to the nation's honey industry, ranking
first in the nation for honey yield per colony and fifth for total yield.
Montana honey is premium grade white "clear" honey and is used to upgrade honey
142
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from other regions. Approximately eight percent (6,600) of the honeybee
colonies of Montana are located in our study area. These colonies produce about
739,200 pounds of honey annually (based on state average yield), with a
wholesale value of $332,640 and a retail value of $724,416 (personal
communication, W. Kissinger, State Apiarist). But estimates of the economic
value of honeybees as pollinators range from 20 to 100 times their wholesale
value as honey and wax makers (Winski, 1974; E. McGregor, 1976; F. Moeller,
personal communication). Using these figures, it appears that the honeybees of
southeastern Montana are worth from $6.6 million to $33 million annually just as
pollinators, excluding their value to California growers.
LITERATURE REVIEW
Insects and other invertebrate animals seldom are studied in air pollution
investigations, yet the available literature suggests significant interactions
of insects and pollutants. Sulfur oxides, nitrogen oxides, carbon oxides,
particulates, hydrocarbons, fly ash, acid mists, and numerous trace elements
impact entomological systems (bibliography in Appendix A). For example,
Hillmann and Benton (1972) conducted investigations near a 615 MW coal-burning
generator in central Pennsylvania and observed several consequences to insect
populations which were linked to sulfur dioxide air pollution, while a European
study associated honeybee mortalities with arsenic emitted from a coal-fired
power plant, 6 km distant (Ferencik, 1961).
Air contaminants may accumulate in the tissues of insects by ingestion,
respiration, or penetration through the integument (Debackere, 1972). Toxic
substances may be passed along the food chain and accumulate in predatory
insects, resulting in host-parasite or host-predator imbalances (Dewey, 1972;
Hillmann, 1972; Hay, 1975). In addition, pollinating insects, especially the
social insects, appear to be very susceptible to toxicosis from atmospheric
pollutants (Dewey, 1972; Knowlton e_t a]., 1950; Bromenshenk and Carlson, 1975).
Social insects such as domestic and wild bees forage over large distances,
transport materials back to the colony and store them as food supplies. These
insects are morphologically and behaviorally specialized for the collection and
transport of small particles (i.e., pollen) which may lead to the inadvertent
accumulation of small, particulate contaminants.
Studies of pesticides and of anesthesia with carbon dioxide and nitrous
oxides indicate that the effects of toxic substances on insect physiology,
biochemistry, and behavior include sublethal effects, such as disorientation,
memory loss, and permanent changes in behavior as well as the more obvious
lethal effects (Schricker and Stephens, 1970; Ribbands, 1950). Gerdes et al_.
(1971a and 1971b) found that 1.3 to 2.9 ppm of hydrogen fluoride reduced
hatchability and fecundity of fruit flies, Drosophila melanogaster, probably as
a result of genetic damage, and Ramel (1967) induced chromosome disjunction by
feeding D. melanogaster 0.25 mg mercury. Recently, European investigators
(Debackere, 1972; Toshkov et aj. , 1974) concluded that some insect species,
particularly honeybees, have a very high potential as sensitive and efficient
indicators of toxic substances and acidic solutions.
Numerous reports indicate that "trees weakened or injured by pollutants are
more likely to be attacked by insects that normally require weakened trees for
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successful reproduction" (Haegle, 1973). The number of arthropod pests that
have been associated with pollution-predisposed trees is small. However,
certain sucking insects or those that feed in concealed places, such as under
bark, are more likely to proliferate, while foliage-feeding insects are more
subject to poisoning because of their greater exposure both to airborne toxins
and through feeding (Hay, 1975). Two questions repeatedly are raised in reviews
of the subject (Haegle, 1973; Hay, 1975; Ciesla, 1975): Are trees predisposed
to arthropod attack and damage because of air pollution stress? Do air
pollutants affect the predators and parasites of a pest arthropod and, as such,
affect important regulators of pest population numbers? These questions are
particularly important in view of the conclusion of Mattson and Addy (1975) that
phytophagous insects act as regulators of primary production and nutrient
cycling of forests and thus have long-term interactions with fundamental
ecosystem processes.
MATERIALS AND METHODS
The techniques and materials employed for the entomological investiga-
tions, including the monitoring and analysis for the accumulation of sulfur and
fluoride in insects, are lengthy and have been published previously (EPA, 1976;
Gordon et al_. , 1976; Kay et al_. , 1975; Carlson et a_L , 1974). Summaries of these
procedures, modifications, deletions, and additions are indicated under the
reports on specific objectives.
OBJECTIVE #1
SELECTION AND ESTABLISHMENT OF PERMANENT STUDY SITES
DISTRIBUTED SO THAT GRADIENT EXPOSURES AND RESPONSES TO AIR
POLLUTANTS FROM THE COLSTRIP POWER PLANTS OCCUR
In September of 1976, participants from federal agencies and research
institutions convened to examine the role of applied ecology in environmental
problem solving (Johnson, 1976). They concluded that post-action impact
monitoring programs are potentially valuable aids to the improvement of modeling
and predictive capabilities and provide valuable opportunities to measure the
results of controlled experimental perturbations of diverse ecosystems. They
stressed the need for an experimental design that could identify unforeseen or
underestimated impacts, provide a base against which to compare post-action
impacts, improve future predictions, and be amenable to information transfer.
We believe that plot selection is a critical element of pre- and post-
action impact evaluations. First, any controlled study of perturbations in this
type of field investigation depends on plot distribution which in turn should
consider known or suspected wind channels from the stationary source and the
degree of exposure of the plot to contaminants emitted by the source. For
example, because fluorides are transported in the atmosphere, vegetation on
ridges or elevations downwind from the emission source is more likely to be
subjected to fluoride stress than vegetation upwind from the source or in
valleys (Carlson and Dewey, 1971j. Second, air pollution impacts may be short-
term and acute or long-term and chronic. The latter may be discernible only
after a period of several years. Therefore, for maximum usefulness, study plots
144
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should be relatively permanent as regards location and accessibility, which is
why plots were located on public domain whenever possible.
For the Colstrip studies, permanent study plots were established in the
ponderosa pine-skunkbush and the cool season-short grass ecosystems. Ponderosa
pine-skunkbush sites were located at varying distances from Colstrip in eight
directions. General locations were determined by plotting intersects of eight
compass radii (45.0 degree sectors) on a topographic map, with concentric
circles representing a two-fold geometric progression of distance (8 to 120 km)
from the power plants. Final site selection was made in the field based on these
criteria: (1) Each site should be on public domain whenever possible; (2) each
should be on the most elevated and exposed terrain in the immediate area, and
(3) each should be composed of ponderosa pines of different ages associated with
a diversity of understory species.
Prior to the initiation of this project, we conducted baseline studies for
the state of Montana at 35 pine sites in the Colstrip area (DNRC, 1974). From
these sites, 14 were chosen for use in the pre-operational phase of the
entomological studies for EPA and three sites were added (3 to 5 km from
Colstrip) where extensive air monitoring was being conducted by the state of
Montana and by Battelle Northwest Laboratories. The majority of these sites
were located in the path of the prevailing winds, downwind from the stacks
(Figure 5.1A, locations published by EPA, 1976).
Twenty-six commercial apiaries (Figure 5.IB, locations in Appendix B) were
established at distances of 6.7 to 130 km from Colstrip as permanent plots in
the short grass ecosystems and bordering agricultural lands. Apiaries of
southeastern Montana are situated along river and creek floodplains and, as
such, are in the valleys. Most of the beeyards within 20 km of Colstrip are
along Rosebud Creek; only three are located elsewhere. The choice of bee sites
was limited to existing apiaries, which tend to be permanent because of
Montana's legal restrictions on location. Four men own and manage all of the
6,600 commercial bee colonies in the study area. They all are interested in the
results of the study and have given permission to obtain samples from their
hives at any time and from any location. Thus, we believe that we shall continue
to have access to these sites and resources.
To date, we have one of the most extensive field plot systems in south-
eastern Montana. In addition to the EPA sites, similar study sites have been
established for use in related studies sponsored by other state and federal
agencies (USFS, ERDA, State of Montana). There is baseline data from 50
ponderosa pine-skunkbush sites, 26 apiary locations, and six grassland
enclosures, and entomological studies are being conducted at 17 of the pine
sites, all of the apiaries, and on the two ZAPS grassland exclosures.
The initial results in the post-operational investigations at these sites
confirm the belief that we not only have established an extensive network of
monitoring points but that these points are being subjected to differential
levels of air pollution (see Objective #5 and section by Gordon et al_. ).
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Figure 5.1. Colstrip study area; locations of (A) ponderosa pine-skunkbush and
(B) apiary sampling sites.
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OBJECTIVE #2
COMPILING INVENTORIES OF INSECT POPULATIONS, INFESTATIONS,
AND DAMAGE TO INDIGENOUS PLANT SPECIES AT THE STUDY
SITES TO PROVIDE A BASIS FOR THE SELECTION OF MORE
SPECIFIC GROUPS FOR USE AS INDICATOR SPECIES
These surveys wtfi Inltittid at thi ponderost plne-skunkbush sites to
dettrmlne thu Insect ftuna pristnt, to ditirmlnt thi stitus of Insist pests of
pondirosa pint and thilr potential for outbreak, ind to dittrmlnt ths itatus of
binsfldil Insicts, such 11 Initeti prtdidous on folligt fisdtri. It wis
hypothiilnd that this 1nformit1on would provide a bisls for thi itlietlon of
men spidfle groups of Instcti as 1nd1e§tor iptclti, ts well is provldi
supplement!! 1nformit1en, such is hibltat Information ind distribution records.
Wt did not Inttnd to expend largs amounts of study resources amassing extensive
species lists or collecting quantitative data such ts estlmatss of absolute
population size or diversity Indices, It was felt that the natural variability
of many Insect populations precluded or greatly reduced the probability of using
this type of data for meaningful post-action evaluations of pollution-induced
perturbations or of separating these perturbations from the interaction of other
physical and biotic variables.
Investigators from the Colorado Natural Resources Ecology Laboratory
(CNREL) attempted to document changes in total arthropod population numbers and
biomass on grassland sites using techniques that had successfully shown
responses of arthropods to artificially-induced environmental stresses such as
moisture and grazing pressure. They encountered problems applying this approach
at the EPA S02 fumigation plots because of low population numbers and high
natural variability. However, their extensive insect sampling provided species
inventories for the grassland ecosystems; these inventories allowed us to
concentrate our attention on the pine ecosystems.
Several techniques were used to conduct insect inventories: (1) Exami-
nation of state and federal entomological collections (State Department of
Forestry and USDA Forest Service, Region 1) for identified species collected
from southeastern Montana timber stands; (2) searches of records and collections
pertaining to insects and insect outbreaks in ponderosa pines of eastern
Montana; (3) on-site observations and sampling procedures (Appendix C)
conducted in August and October, 1974, .and once every two or three weeks from
May 1 through October 15, 1975; (4) examination of insects in or on foliage
samples obtained from all of the ponderosa pine sites, both from the EPA studies
and those from the Forest Service project, and (5) utilization of sticky traps
(after Williams, 1973) which consisted of 35 x 45 cm rectangles of 1.5 ml
aluminum coated with Tack-Trap®and fastened 3.5 meters above the ground to the
trunks of each of five trees at five locations. The sticky traps were changed
every six weeks from May 5 through November 20, 1976. The purpose of this
trapping, begun in 1976, was to trace the phenology of some of the major or
dominant insects on pines and, hopefully, to obtain specimens of ephemeral adult
forms, which were necessary for the identification of a few species which we had
seen only in immature stages. Results from the sticky traps were not complete at
the time (March 1, 1977) of preparation of this report.
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All insect specimens were labeled according to site, date of collection,
and indexed by number in a log containing comments. The specimens were pre-
served in 80 percent ethanol in glass vials until they could be identified.
The emphasis for this part of the entomological investigations has been to
provide supportive data for the ponderosa growth-health studies (see section by
Gordon et aJL for study plan and results). The growth-health studies quantify
pathologies occurring on pine foliage obtained from ten permanently marked trees
at each of the plots. Foliage samples were taken from upper and lower crown
positions on the side of the trees facing Colstrip. The foliage was then
subjected to analysis of sulfur and fluoride content, and the percent occurrence
of ten needle pathologies (e.g., insect, fungal, pollution damage), the percent
needle retention, needle length and cross-sectional area, and chlorophyll
content. Pathology determinations were made for each group of 100 needles from
the preceding four years of growth. The lengthy procedures have been published
(Gordon et aJL , 1976); procedures and results are summarized in the section by
Gordon et aJL of this report. These procedures, including sample size and data
handling, were evaluated by PEDCO Environmental Specialists, Cincinnati, Ohio,
and their recommendations regarding the study plan have been incorporated.
An insect checklist is included in Appendix D. On-site measurements and
evaluations dealing with tree physiology and insect-damaged portions of trees
other than foliage are summarized in Appendix E. The most frequent and serious
insect damage syndromes are illustrated in Figures 5.2 and 5.3.
From our pre-operational studies, the following conclusions were drawn
regarding the status of insect species present at the ponderosa pine-skunkbush
plots:
(1) Compared to most stands of ponderosa pine in Montana, trees on the
study plots in southeastern Montana were exceptionally healthy and generally
demonstrated little or almost no insect, disease, or pollution damage, except in
a few localized areas.
(2) The primary causes of tree mortality appeared to be porcupine
girdling, storm breakage and blow-down, and fire. Insects and disease as
primary causes of death were rare, except on the Cheyenne Indian Reservation
which has had a history of occasional insect outbreaks in timber stands and
continues to have some localized insect problems.
(3) Insect populations were generally low and insect damage was minimal.
(4) The most prevalent destructive insects and damage included: (a) Pine
cone boring by the larvae of beetles and moths (Conophthorus, Laspeyresia, and
Dioryctria spp.), which occurs on all plots and occasionally destroys
significant numbers of cones and seeds; (b) scale insects (Matsucoccus secretus
Morrison) found on most trees under the fascicular sheaths or between the
needles in two year or older needles; (c) foliage feeding by several weevils
such as Scythropus elegans (Couper) observed on all plots and Magdalis sp.
observed on new growth at several plots; (d) pine loopers (Phaeoura mexicanaria
(Grote)) seen at six plots but in very low numbers; (e) several sawflies
(Neodiprion spp.) seen feeding on foliage at several plots but only on
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Figure 5.2. A. Ponderosa pine killed by bark beetles (Dendroctonui fionderosae). B. Young P°"derosa
pine girdled by a porcupine, which will cause the tree to die. C. Immature pine cone of
ponderosa pine attacked by Conophthorus pondgrosae Hopk. The cone is dwarfed by the attack
of the insect. D. A pine seed moth la^T^oWbly Lasjjorresia fiiprana (Kearf.). working
through the pith into seeds. The larval stage of three species of pine seed moths are
indistinguishable.
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Figure 5.3. A. Basal portion of a needle of Pinus ponderosa showing the necrotic symptom of damage
found under the fascicular sheath. B. An infestation of pine needle scales (Phenacaspis
pinifoliae) on ponderosa pine. C. Notches chewed in the needles of Pinus ponderosa by
weevils such as those of the genus Scythropus. D. A cluster of larvae of a species of
Neodiprion (sawfly) feeding on needles of Pinus ponderosa. E. Close-up of needles of
Pinus ponderosa damaged by a pine needle miner. A larva is inside the needle. F. Short-
ened needles with bulbous bases characteristic of attack by several pine gall midges, family
Cecidomyiidae. Many of the western species have not yet been named.
150
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occasional branches; (f) infrequent, localized infestations of bark beetles
(Dendroctonus ponderosae Hopk. and Dendroctonus brevicomi LeC.) and engraver
beetles (Ips cal1igraphus (Germar) and !_. pini (Say)), and (g) ubiquitous
populations of aphids.
(5) The most prevalent insects during the summer months were lady bird
beetles (particularly Cleis sp. and Neomysia sp.) and a leaf-footed plant bug
(Leptoglossus sp.), which was common in the autumn months. The beetles are
predacious and are reported to feed on a variety of foliage feeders such as
mites,, scale insects, and aphids.
(6) Many other insect predators, parasites, and parasitoids prey on the
harmful insect species. Particularly conspicuous beneficial insects were
lacewings of the family Chrysopidae and Mantispidae, several Icheumonids
(parasitic Hymenoptera), and beetles of the families Cleridae and
Coccinellidae. In addition, a profusion of dead aphids exhibiting large exit
holes in the abdominal region indicated the interaction of an unidentified but
prevalent parasite.
(7) The potential for an immediate epidemic of destructive insects was
considered low except in a few localized areas. The pine looper, Phaeoura
mexicanaria, occurred in epidemic numbers on the Northern Cheyenne Reservation
and the Custer National Forest in 1969 and 1970 (Dewey, 1970). About 20,000
acres were heavily defoliated and another 43,000 were moderately to lightly
defoliated. The defoliation appeared to predispose many trees to attack by bark
beetles (Ips cal 1 igraphus, 1^. pini, and Dendroctonus valens LeC.). A collapse
of the pine looper population in 1970 was attributed to the spread of a
pathogenic bacterium.
M. McGregor, leader of the Bark Beetle Evaluation and Control Group, USFS,
Region 1, inspected several Col strip pine sites during August 3-8, 1975, and his
report (in Gordon e_t a!. , 1976) closely agreed with our own observations.
McGregor conc]uded that a weakening of stands might lead to another population
build-up of the pine looper (£. mexicanaria) but discerned no immediate
potential for outbreak. He noted infestations of the mountain pine beetle (D.
ponderosae) and the pine engraver beetle (!_. pini) in a blow-down at one plot in
the Custer National Forest and in areas of thinning of second-year growth of
ponderosa pine in the Three-Mile Creek area, which had a potential for a
localized build-up by 1976.
Besides providing species lists and information on the pre-operational
status of insects associated with ponderosa pines, these insect surveys may be
extremely useful for post-impact monitoring. Rather than attempting to directly
quantify insect population dynamics, we are monitoring the insect damage
incurred by foliage by assessing the percent needles attacked or consumed by
insects. In his book on ecological methods as applied to insect populations,
Southwood (1975) considered estimates of the amount of a plant consumed,
expressed either as a quantity of material consumed or as some index of damage,
to be the most meaningful biological measure of the effect of an insect
population-on a plant stand, although these estimates are only an approximate
index of the size of the insect population. Damage per se is of more intrinsic
interest to us than absolute insect numbers because insect damage affects growth
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rate and is reflected by growth rings (Mott et al_. , 1957) and because increased
insect damage to conifers has often been associated with pollution-stressed
trees (review by Haegle, 1973). However, in every case, reports of tree
mortality or decreased production due to insect attack on pollution-stressed
trees relied on studies conducted after the stationary source(s) had been in
existence for some period of time. Generally, there was little, if any,
documentation of insect conditions prior to the operation of the source. Thus,
if an insect outbreak occurred, the argument often was made that the outbreak
was a natural part of the life cycle of the pest and was a random or coincidental
occurrence rather than the result of increased levels of a toxic pollutant.
In the Col strip area, we quantified foliage damage caused by several agents
including insects (see section by Gordon et aK ) prior to the operation of the
power plants, determined the current status of insects known to be destructive
to ponderosa pine, and obtained records concerning the history of insect
problems in the area on the nearby commercially usable stands of timber, the
Custer National Forest and the Indian reservations. We consider the area to be
free of serious insect problems, as evidenced both by records and by
observations of the tree stands at each site. However, some insect species were
identified which appeared to be of particular interest to observed foliage
damage and post-impact responses. These are discussed under Objectives #3 and
#4.
OBJECTIVE #3
ANALYSIS AND SELECTION OF INSECT POPULATIONS WHICH HAVE A DIVERSIFIED
BUT UNDERSTANDABLE INTERRELATIONSHIP WITH INDIGENOUS PLANT SPECIES
AT THE STUDY SITES IN ORDER TO EXAMINE THE EFFECTS OF AIR
CONTAMINANTS ON SYMBIOTIC OR COEVOLVED SYSTEMS
Inventories (USDA Rangeland Laboratory, Bozeman, Montana; USFS, Region 1,
Missoula, Montana; Environmental Studies Laboratory, University of Montana,
Missoula) of the pine and grassland ecosystems of southeastern Montana indica-
ted that a minimum of 1,500 to 2,000 species of insects commonly are associ-
ated with the indigenous plant species. We elected to concentrate our efforts
on a few specific life systems, an approach which is believed to yield good
quality control and the opportunity to test feasible hypotheses. A "life
system" is defined as the part of an ecosystem which determines the existence,
abundance, and evolution of a particular population. According to this con-
cept, the inherited properties (array of genotypes) of a population*transform
environmental resources of matter and energy into phenotypes or the individ-
uals of the species. These individuals form populations with group character-
istics (e.g., birth rates, dispersal, mortality). The continuance and abun-
dance of a population are determined by interactions between inherited charac-
teristics of the subject and essential attributes of its environment. The
intrinsic qualities of the subject species and of its environment are termed
the co-determinants of population numbers. The co-determinants control
population performance (e.g., reproductive capacity, mortality, immigration,
and emigration or, in other words, numbers and persistence). The manifes-
tations of determinant processes are certain ecological events, including
primary demographic events such as matings, birth rates, mortalities, densi-
ties, and distributions and secondary events which affect or modify the primary
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occurrences such as resource availability and inimical agents, both biotic and
abiotic. This life system approach to the ecology of insect populations was
formulated and presented by Clark et al_. , 1967.
Having decided to pursue a life system approach, a list of 28 insect
species (EPA, 1976) was drawn which were thought to be: (1) Major components of
the pine and grassland ecosystems of the study area; (2) dominant and/or useful
indicators or estimators of pollution; (3) economically important, and (4)
closely coevolved with dominant, indigenous plant systems. The rationale for
our emphasis on the latter point was expressed in a recent symposium on the
coevolution of plants and animals (Gilbert and Raven, 1973). They stated in the
introduction:
(Revolutionary relationships are, by definition, the product of
historical change Devolution); yet this historical change is still
proceeding. The relationships between elements in a contemporary
ecosystem are dynamic in both an evolutionary and in an ecological
sense, and depending upon the generation times involved, may even
change very rapidly during a given period of observation. This is all
the more likely in these times of extensive environmental pollution
and the consequent alteration of relationships within ecosystems that
must affect all of our observations.
Tightly structured organism-organism relationships hold
considerable promise for the understanding of the area as a whole.
To test the hypothesis that tightly structured insect/plant interactions
may change rapidly as a consequence of pollution-induced alterations of these
relationships within ecosystems, we selected from the list of 28 species a few
herbivorous insects of ponderosa pines (Objective #4), insect pollinators and
the plants from which they obtain their nectar and pollen (Objective #5), and
ground-dwelling beetles which act as scavengers and decomposers and return
nutrients to the soil ecosystems, which in turn affects plant growth.
OBJECTIVE #4
SELECTION AND TESTING OF INJURY- AND DISEASE-CAUSING
INSECTS TO BE UTILIZED FOR MORE INTENSIVE STUDY
The original objective was to select and pre-test, for use at the study
sites, insect/plant systems which not only appeared to be of greatest interest
to project goals but also would be easy to manipulate. Towards this goal, we
reared (EPA, 1976) laboratory populations of grasshoppers (Melanoplus
bivittatus) and several herbivorous pine insects such as the pine needle scale,
(Phenacaspis pinifoliae (Fitch)), bark beetles (Ips and Dendroctonus spp.), and
aphids (Cinara spp.). In general, the insects were provided with their
preferred food such as boughs of foliage for needle feeders, bolts of wood for
bark beetles, and native grasses for grasshoppers. Work was terminated with
grasshoppers because of the interest and subsequent work with these insects by
the investigators from Colorado (CNREL) and also because five years of personal
experience with the grasshopper systems of Montana's rangelands revealed that
the population dynamics and host relationships of these organisms were extremely
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complex and that population changes were unpredictable. In addition, a study by
Dewey (1972) indicated that grasshoppers near an aluminum smelter accumulated
less toxic materials (fluoride) than pollinators, predators, or many other
foliage feeders.
Inventories, examinations, and measurements of foliage pathologies
revealed the presence of a scale insect (identified as Matsucoccus secretus
Morrison) found under the fascicular sheath of needles, occurring often on or
near a zone of basal necrosis (Figure 5.4) in foliage generally at least two
years or older. It is not known at present if this insect is a primary factor, a
secondary invader, or a coincidental occupant in the production of all or a
portion of this necrosis. The basal necrosis is similar to that which Dr.
Gordon has produced in the Environmental Studies (EVST) greenhouse by "acid
mists." Therefore, it is felt that it is essential to clarify the relationship
of this insect to the necrosis and that the implications of this relationship
are far reaching. Also, Siewniak (1971) reports that Matsucoccus pini occurs
most frequently on Scotch pine in areas with polluted air. For these reasons,
highest priority is being given to a study of the lifecycle and relations of
this insect to ponderosa pine.
This insect first was observed in 1974 but was seen only occasionally.
Examination of foliage samples in autumn of 1975 indicated that the insect was
more common than it was thought, probably because, as Keen (1952) states, they
are extremely difficult to detect because they push their way under the
fascicular sheath and may take on the color of their surroundings.
A specialist, Dr. Douglas R. Miller (USDA Agricultural Research Service,
Beltsville, Maryland), verified the identity of the insect. The insects are
small, less than 1.5 mm long, and inconspicuous. The sheath must be dissected
from the needle in order to find the insect; if the sheath is pulled away from
the needle, the insect is removed with the sheath.
Macroscopic and microscopic examination of needle bases disclosed that the
basal necrosis which occurs beneath or just above the fascicular sheath appears
to grade, in degree of damage, from a pale or dark brown blemish on the surface
of the needles to a small localized enlargement (blister) of the mesophyll
tissues; this pushes the hypodermal and epidermal tissues out into a blister.
An eruption or splitting of the epidermal and hypodermal tissues may occur,
usually in association with the blistering. The original hypothesis,^which has
since been discarded, was that there were two distinct needle tissue disease
syndromes; the first was termed basal necrosis for the purpose of needle
pathologies, the second was termed basal scale. The reason for the latter
terminology was that cases were noticed in which several insects, especially the
basal scale, were near or in the areas of tissue eruption or hypertrophy. Based
on continued histological and macroscopic examinations of the insect and of this
disease, it is now believed that there is no real difference between these
damage syndromes except of degree.
An approach towards obtaining an understanding and a working knowledge of
the basal scale insect involves several procedures. In order to determine the
extent of the distribution of the insect, our laboratory staff has been
recording the presence or absence of the insect and the life stage of the insect
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Figure 5.4. Scale insect (Matsucoccus secretus) and the hypertrophied or "blistered" needle base which
is vulnerable to attack by insects and saprophytic fungi. The fascicular sheaths of the
needles (Pinus ponderosa) have been removed.
A. Dark-field photomicrograph of an unidentified insect found under the fasicular sheath.
B. Cross-section of erupted needle. C. Scales in permanent feeding position on a
needle base. D. Motile stage of scale, or possibly of an adelgid, in temporary feeding
position near erupted tissues of a needle.
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insect on each sample of TOO needles/tree/plot used to quantify the various
types of needle injury. Examination of the needle bases was facilitated by the
use of a 70-power zoom objective binocular dissecting scope (Bausch and Lomb).
In 1976 there was a search for this insect at all of the plots for all of the
projects (EPA, ERDA, USFS) or on a minimum of 500,000 needles from 22 plots from
mid-March through mid-September. Representative specimens of the different
pathology types and of the different stages of the insect were prepared for
histological analyses, killed and fixed in FAA(formalin-aceto-alcohol),
sectioned by a paraffin technique (Johansen, 1940), and stained either with
Fuelgin's Fast Green Schedule or a modified Fleming's triple satin method (Raske
and Hodson, 1964). The modified Fleming triple stain was used to differentiate
insect feeding tracts; using this procedure, xylem stains blue, phloem orange,
and mucoprotein secretions from feeding red. Observations of sections and
photographs were performed using a Reichert "Zeptopan" microscope with bright
field, dark field, and phase optics.
The results to date concerning the prevalence of the insect are summarized
in Table 5.1. The insect appears to be common in the southeastern Montana area,
including Billings, occurs in relatively low numbers, and seems to prefer
younger trees. Macroscopic and microscopic examination indicated that the
motile adults and larvae frequently were entrapped by exudates from erupted
needle tissues and that feeding often occurred adjacent or proximal to the areas
of basal necrosis, but not necessarily in the damage zone. In fact, most of the
larvae in permanent feeding positions were on healthy-appearing parts of the
needle. This, in conjunction with observations by Dr. Gordon that the basal
damage probably occurs before the meristematic tissue is fully differentiated,
indicates that the insect may not cause the damage but may be attracted to the
area of tissue damage.
TABLE 5.1. PERCENT OCCURRENCE OF TOTAL BASAL NEEDLE NECROSIS AND OF BASAL
SCALE INSECTS ON NEEDLES OF PONDEROSA PINE AT SIX SITES NEAR
COLSTRIP AND BILLINGS, MONTANA - 1976.
Year
of
Foliage
75
74
73
72
Number of
Foliage Samples
(trees)
48
48
46
10
Number of
Needles
Dissected
2611
2742
2514
461
Percent
Non-necrotic
Needle Bases
> 98
81.6
50.8
34.7
Percent
Necrotic
Needle Bases
< 2
*
18.4
49.2
65.3
Percent
Basal
Insects
< .5
4.9
7.5
13.9
(bibliography, Appen-
damage to pines over
A review of the literature concerning Matsucoccus
dix F) revealed that some species have caused severe
extensive areas and that some forms cause needle galls; but the damage syndrome
discerned in- this study is not characteristic of any reported damage attributed
to Matsucoccus. Histological comparisons of the feeding patterns of these
156
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organisms in relation to feeding and damage by other insects (Figures 5.5 and
5.6), fungi, and abiotic factors such as frost, drought, and acid mist should
clarify the role of this insect. There are Matsucoccus infested trees in a
greenhouse at Missoula and these are being used as a source of basal scale for
further studies of life cycles, feeding, and damage. We hope to establish a
suitable population for inoculation tests based on Koch's Postulates.
OBJECTIVE #5
SELECTION AND TESTING OF BENEFICIAL INSECTS
TO BE UTILIZED FOR MORE INTENSIVE STUDY
The previous objective emphasized coevolved systems of insects and plants
in which the insects were detrimental both in terms of harm to the plant and in
subsequent economic loss because of plant damage. However, there are many
coevolved insect/plant systems in which the insect is beneficial. The most
familiar aspect of advantageous insect-plant interaction is the pollinator
relationship. Honeybees were chosen as the subject species because of their
known sensitivity to pollution stress (Toshkov et al., 1973), ecologic and
economic importance, availability (in terms of numbers"), and manageability. It
is believed that they will be indicative of the population responses of other
pollinators, and to test this hypothesis honeybees and other pollinators are
being observed at the S02 fumigation plots for comparative purposes. The native
bees, especially the physically smaller species, may be much more susceptible to
zootoxins than honeybees (Johansen, 1972).
We would like to work with predatory and predacious insects that attack
forest pests and with the predatory ants of the grasslands; but the initial pre-
testing indicated that this exceeded study resources, necessitating priority
assignments. In 1976, studies of ground-dwelling beetles were added at the ZAPS
plots on the basis of previous surveys and biomass studies by the Colorado
investigators (CNREL).
While behavioral responses of honeybees to controlled exposures of S02 have
been investigated at ZAPS, the studies near Colstrip have concentrated on the
use of honeybees (Apis mel1 ifera L.) as accumulators and indicators of the
extent and magnitude of the phytotoxic and zootoxic effluents from the Colstrip
power plants (see Objective #6).
OBJECTIVE #6
CHEMICAL ANALYSIS TO DETERMINE SULFUR AND FLUORIDE CONTENT
OF INDIGENOUS PLANTS AND INSECTS TO ESTABLISH BASELINE
LEVELS AND TO MONITOR ANY ACCUMULATION OF THESE MATERIALS
WHICH ARE RELEASED BY THE CONVERSION OF COAL TO ELECTRICITY
As mentioned previously, it was hypothesized that honeybees would be useful
in the collection and detection of toxic materials from the environment.
Numerous reports supported this hypothesis, although many were based on studies
in Europe during the 1950's (review by Debackere, 1972). The usefulness of the
published information from these studies is sometimes questionable because of
two problems. One is the lack of a standard expressing the concentrations of
157
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t >/.,,-ir .» . >•• •-- *
rf>) V:"^'/ ^
Figure 5.5. Base of needle and fascicular sheath.
A and B. Longitudinal sections of needle showing an unidentified
mite under the sheath. C and D. Cross-sections of needles and
sheath.
158
-------
Figure 5.6. Weevil damage to needles of Pinus ponderosa.
A and B. Cross-sections of damaged needles and insect.
D. Longitudinal sections of weevil damaged needles.
C and
159
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toxic substances in insects and in the vegetation they forage. Levels are
reported as g/insect, g/kg, mg%, ppm dry weight, ppm wet weight, and similar
designations so vague that one cannot determine the basic unit of measurement.
The other problem is the assumption that data which correlates changes in
insect populations, such as mortalities with tissue concentrations of a single
contaminant, may be used to estimate toxicity or harm. This type of data may be
sufficient to demonstrate harm by air pollutants but does not identify the
specific agent and ignores synergistic effects. In addition, these studies were
conducted near industrial sources with sites distant from the source used as
controls. These controls were designated as such because of their distance from
the source. But the acid rain literature indicates that some forms of air
contaminants may impact areas as far as 500 km from the source, and if the air
pollution from the source was widespread, these controls may not have reflected
pre-operational baseline levels of pollutants.
Since the initiation of the EPA studies in August 1974, we have acquired
quantitative analyses of fluorides, sulfurs, and pesticides in adult worker
honeybees, their pollen and honey food supplies, and their water supplies.
Bees, honey, and pollen were obtained from commercial apiaries at 26 locations.
Twelve of the apiaries are within 20 km of Col strip, are utilized as primary
sites, and have been sampled eight different times during the last two years.
The most distant beeyards are used as (controls) and as indicators of long range
pollutant transport. Individual samples of bees were taken from at least four
different hives, and a combination sample was collected from a minimum of eight
to twelve hives at each location. Individual honey and pollen samples were
pooled from several cells within a colony. Individual bee samples were defined
as 200 to 300 adult worker bees taken from the same colony. Combined bee samples
consisted of 150 to 300 bees per colony taken from each of eight to twelve
colonies and grouped together at the time of collection. Individual samples
served as a measure of variation among colonies; the combined samples served as
an average sample, allowed comparison with the individual samples, and provided
a sufficient number of bees for those analytical procedures, such as pesticide
analyses, which require a large number of bees (100 g or more wet weight). A
100 g sample of bees taken from a single colony could significantly affect the
field work force and as such affect production.
Apiaries in southeastern Montana are laid out in rows of wooden pallets,
each pallet supporting four to six colonies (Figure 5.7A). The selection cri-
teria for hives consisted of sampling all suitable hives in each quadrant
established by the rows of pallets. A suitable hive was one in gofld physical
condition (e.g., paint not peeling, supers and covers intact and properly
positioned, clean appearance), free of obstructions and contaminants (e.g.,
vegetation and dirt at the entrance) and containing a strong colony (e.g.,
vigorous, intense flight activity, populous). Four individual bee samples, one
from each quadrant, and a combined sample were taken at each apiary during each
sample period in 1975 and 1976, excluding the spring periods.
An acrylic plastic vacuum apparatus (Figure 5.7B) was developed for
collecting bees at the entrance to a hive to minimize inadvertent contamination
at the time of collection. All samples for sulfur and fluoride analysis were
frozen immediately by bagging them in plastic Whirl Paks® and placing them under
a block of dry ice in a styrofoam cooler. Since pesticide determinations may be
160
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Straight Venturi
Inlet (2 cm I.D.)
Exhaust Outlet
(5.2 cm I.D.)
Air
Bee Barrier
(3 mm
perforations)
Figure 5.7. A. Apiary at SU-3 by Rosebud Creek near Colstrip, Montana. B. Acrylic aspirator used to
obtain honeybees for chemical analyses. A battery (12 volt) powered squirrel cage fan pro-
vides a vacuum and is connected to the exhaust outlet via a flexible plastic hose. Air and
bees are drawn through the venturi inlet; the bees are deposited in the collection jar. The
jar is removed by unscrewing it from the aspirator, giving a sharp downward rap to force the
bees to the bottom, and quickly removing and capping the jar.
161
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confounded by residues from plastics, we used a metal vacuum collector, based on
the design of the acrylic apparatus, and stored the frozen samples in new one-
pound glass honey jars. Honey was collected by scraping a teflon-coated spoon
across the surface of the comb. Pollen was removed from cells by the use of a
narrow plastic pick. Samples were stored at -10°C until they could be prepared
for analysis.
Clean sampling and storage equipment was of the highest priority. Cleaning
procedures were determined by the analyses to be performed. All plastic
equipment, except the clean, sterile Whirl Paks® was cleaned by washing in
detergent and tap water, rinsing with 1:1 nitric acid, then with tap water,
followed by a one normal rinse with hydrochloric acid, then with tap water, and
a final rinse with distilled and deionized water (Taras et a_L , 1971). The
equipment was air-dried, face or mouth downward, on a plastic pad. Glass and
metal equipment was cleaned by washing in detergent and tap water, rinsing with
distilled and deionized water, and then rinsing with acetone followed by a rinse
with hexane. Bottles were air-dried, mouth downward, on an aluminum foil sheet.
The mouths of all glass jars were sealed with clean aluminum foil to exclude any
impurities adhering to the inside of the covers (procedures recommended by the
Beltsville Pesticides Laboratory). All chemicals utilized for cleaning were
reagent grade.
Preparation for fluoride and sulfur analyses consisted of drying and
grinding whole bees and subsequent analyses using an Orion specific ion elec-
trode for fluoride and a Leco induction furnace for sulfur determination (EPA,
1976). Pesticide analyses were conducted by R. Thomas of the EPA Biological
Investigations Laboratory, Beltsville, Maryland. The analytical methodology is
outlined in the FDA Analytical Manual for chlorinated insecticides (e.g., DDT,
DDE, ODD), phosphate pesticides (e.g., malathion, parathion), and carbamate
pesticides (e.g., sevin, carbofuran). FDA methods were sensitive to
approximately 0.01 ppm. Screening procedures utilized gas chromatography, thin
layer chromatography, and gas chromatography in conjunction with mass
spectrometry. In 1975, 700 to 800 bee colonies died from pesticide poisoning in
the study area. Thus, pesticides may be a serious confounding factor.
Choiinesterase analyses were performed courtesy of R. J. Barker, Bee Research
Laboratory, USDA, Tucson, Arizona, using isolation procedures based on thin-
layer chromatography (Winterlin et a_L , 1968) and a chlorimetric determination
of acetylcholinesterase activity (Ellman et aj. , 1961). We have initiated
determinations of trace metals such as arsenic (As), cadmium (Cd), and lead (Pb)
in bees using published procedures for atomic absorption spectrophotfimetry. A
Unicam SP90A Series 2 AA Spectrophotometer coupled to a Van Lab strip chart
recorder was utilized. The results will be published in the next interim
report.
Unless indicated, all statistical evaluations were performed on a Hewlett
Packard 65 calculator or a Dec-10 computer and utilized standard statistical
programs, including mean, standard deviation, standard error, F ratios based on
variances, and Bartlett's test for homogeneity of variances. Significant
differences (.05 confidence level) of variances allowed the use of a t-test for
equality of means with variances unequal, while non-significant differences of
variance allowed the use of a t-test for the equality of means with equal
variances (F ratios and t-tests from Sokal and Rohlf, 1969). All confidence
162
-------
levels were computed for the 95 percent lower and upper confidence limits of the
mean.
Quality assurance tests of the analytical procedures utilized recovery
efficiency studies, and a reciprocal exchange with other laboratories for a
comparison study was carried out by the Boyce Thompson Institute. The
procedures are precise, rapid, and show very good agreement with other methods.
The fluoride probe, based on several years of data, is accurate to ± 0.10 ppm,
while the Leco method for sulfur in bee tissues is accurate to ± 4 percent. The
accuracy of the honeybee analyses and the pollen analyses is very good; however,
the samples must be well mixed after grinding. We did not experience the
difficulties reported by Nation et aj. (1971) who encountered difficulties in
obtaining a homogenous sample of whole bees for the atomic absorption analysis
of major and trace elements in bees, pollen, and royal jelly. Their procedures
were similar to ours except that they ground their bees in a mortar while we used
a Wiley mill which produces a finely ground powder which will pass through a 40
mesh screen. Nation et aj_. concluded that the very hard structures of the head,
antennae, wings, and legs made it virtually impossible to get a homogenous
sample but thought that finely ground materials or analyses of specific body
regions might improve recovery efficiency.
For consistency., all levels of fluorides and sulfurs presented here are
given on the basis of ppm dry weight (pg/g dry weight). To facilitate
comparisons to published reports of levels of contaminants in bees, which are
often expressed as a unit weight per one bee rather than per unit of mass or
weight of bee tissue, we present the results of a series of experiments
conducted to determine the weights of honeybees and the moisture content of bee
tissues (Table 5.2).
The results from the preliminary samples in 1974 are given in Table 5.3.
The mean fluoride content of adult worker bees from 11 sites was 7.4 ppm, while
the mean sulfur content was 4,390 ppm. Carlson and Dewey (1971) reported that
10.5 ppm F was characteristic of bees from a clean or control area in western
Montana, whiTe bees near an aluminum smelter contained 221 ppm F . Obviously
the mean 7.4 ppm F indicated a clean area. In addition, European investigators
have reported levels from 0 to 40 ppm as characteristic of bees from clean areas
(Lillie, 1970). We found only one report of total sulfurs in bees. Hillman
(1972) fumigated colonies with 3.65 ppm S02 for nine weeks and found no
significant differences in sulfur levels between fumigated bees and non-
fumigated adult or larval bees. He reported levels of 3,200 to 4,600 ppm in
adult bees and 1,800 to 3,900 in larval bees.
Our interest in honeybees extended beyond their potential as collectors of
contaminants from the environment. We also wished to discover whether the
levels of these contaminants would increase in bees in the Colstrip area to
levels damaging to the bees and to the bee industry in terms of effects such as
reduced numbers, strength, and vigor, thereby decreasing production. In order
to test the ability to detect contaminants such as fluoride in bees, beekeepers
in Anaconda, Billings, Butte, and Columbia Falls were asked to send samples of
bees from areas known to be subjected to high ambient air concentrations of
pollutants and/or areas in which beekeepers had repeatedly experienced bee
poisonings or unexplained (e.g., pesticides, disease) losses.
163
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TABLE 5.2. WET WEIGHTS, DRY WEIGHTS, AND MOISTURE CONTENT ADULT WORKER
HONEYBEES WITH STANDARD DEVIATIONS.
G Wet Weight/Bee G Dry Weight/Bee Percent Moisture
0.092 ± 0.004* 0.031 ± 0.002 66.85 ± 1.58
0.088 ± 0.006t 0.028 ± 0.002 68.66 ±1.51
t - 66.53 ± 1.95
*Bees collected from ten different apiaries in September 1974 near Colstrip,
Montana. Each value is a mean of ten independent samples of 500 bees. Each
sample of 500 bees represents bees from eight colonies in the same apiary,
5,000 bees total.
tBees collected from 14 different apiaries in August 1976 near Colstrip,
Montana. Each value is a mean of 15 independent samples of 142-216 bees.
Each sample of bees from a separate colony, 2,661 bees total.
tBees collected from 15 apiaries in July and August 1976 near Colstrip,
Montana. Percent moisture based on samples from 64 separate colonies,
approximately 250 bees/sample; 16,000 bees total.
TABLE 5.3. FLUORIDE AND SULFUR LEVELS IN ADULT WORKER HONEYBEES IN PPM DRY
WEIGHT, WITH STANDARD DEVIATIONS*
Site
BN-2
BNE-5
BNE-4
BNE-3
BE-1
BSE-1
BS-1
BS-4
BS-5
BSE-2
BSW-1
Total for All Sites
5.
7.
5.
15.
6.
9.
5.
11.
6.
7.
5.
7.
ppm
2 ±
2 ±
5 ±
7 ±
0 ±
1 ±
1 ±
0 ±
1 ±
0 ±
2 ±
5 ±
F"
0.
0.
0.
0.
0.
1.
0.
0.
0.
0.
0.
3.
57
85
00
21
64
48
14
28
21
35
35
21
ppm
4600 ±
4700 ±
3900 ±
4300 ±
4300 ±
4700 ±
4600 ±
4600 ±
4300 ±
4400 ±
4000 ±
4390 ±
S
282
141
141
141
141
141
282
0.
141
0.
0.
299
00
00
00
*Bees collected near Colstrip, Montana in September of 1974. Each value is
the mean of two independent sub-samples from a combined sample taken from
eight to twelve colonies.
164
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The results are presented in Figure 5.8. It is evident that bees from
these areas had substantially higher fluoride levels than the bees near Colstrip
and the bees from control areas in the study by Carlson and Dewey (1971).
Reportedly, the level of fluoride per bee is a function of activity and varies
with pollution level, geography, season, and the plant species in flower
(Guilhon, 1962). Lillie (1970) stated that the presence of 1 pg F~/bee
indicates the existence of an intermittent or permanent pollution source and
that the lethal_dose of sodium fluoride (NaF) per bee is 4 to 5 pg/bee. Levels
of 4 to 5 ppb F in the ambient air are somewhat toxic to bees (Atkins, 1966).
Based on our estimates of the dry weight of a bee, a level of about 34 ppm would
indicate a pollution source, and approximately 135 to 170 ppm would indicate a
toxic level. The average dry weight of bees from a Florida study (Nation et al. ,
1971) was 32.63 mg, which computes to 33 ppm F as indicative of a source of
fluoride and 123 to 153 ppm as a lethal dose of soluble fluoride.
In 1976 we sampled throughout the growing season, increased the number of
sample locations, sampled individual hives as well as taking combined samples,
and began to collect honey and pollen. Sampling was completed just as the first
Colstrip power plant, Unit 1, began to burn coal in September 1975.
Each year, most of the bee colonies near Colstrip are transported to almond
groves in California where they are rented to pollinate the groves. The bees
are transported from Montana in September or October and are returned in May or
June. Because bees brought back from California may carry contaminants from
that area, colonies were arranged to be left over the winter at apiaries in the
primary study area (within 20 km of Colstrip in 1974, 1975, and 1976). Colonies
were left at six locations in the fall of 1974 (BS-1 , BSE-2, BNE-2, BNE-3, BNE-
4, and BNE-10), at nine locations in 1975 (BS-1, BS-4, BSE-1, BSE-2, BNE-2, BNE-
3, BNE-4, BNE-10, and BN-1), and at one additional location (BS-5) in 1976.
Honeybee colonies in the area southeast of Ashland were never moved from the
region and as such served as further controls.
The presence of pesticides and/or pollutants in migratory bees is a poten-
tial problem. However, the bees are returned with a minimum of food stores and
are in brood boxes. The supers, frames, and other equipment are not taken to
California. If contaminants are carried back from California, they should
decrease as food stores, equipment, and honeybees are resupplied. The studies
indicated that this was the case (Table 5.4). Comparison of fluoride levels in
bees over-wintered in the Colstrip vicinity compared to fluoride levels in bees
brought back to these sites from California demonstrated no significant
difference in fluoride content. All samples in this comparison were collected
after mid-June, by which time contaminants carried from California should have
been eliminated.
In 1975 it was found that one beeyard (BNE-10) had high levels of fluoride
(20 to 108 ppm) in samples from all four collection dates. This same pattern was
repeated in 1976. Prior to 1976, the bees at BNE-10 had not been taken to
California, and the hives had been at that location for at least three years.
Because these colonies had dwindled and were very weak by fall, they were taken
to California in the fall of 1975 and replaced the next spring by strong
colonies; i.e., the apiculturist's breeding stock. As in 1975, the BNE-10
colonies built slowly and produced very little honey in an area that appeared to
165
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100 _
80 -
60 -
40 .
20 -
LI
I
la Ib Ic
4a 4b 5
la,b,c Butte-Anaconda, Montana; near Stauffer Chemical
and Anaconda Smelter
2 Billings, Montana; near Corette Power Plant
3 Rosebud, Montana; fluoride in water supply
4a Columbia Falls, Montana; south of aluminum
reduction facility
4b Columbia Falls, Montana; west of aluminum reduction
facility
5 Col strip, Montana; from 25 apiaries in Southeastern
Montana prior to power plant operation
Figure 5.8. Mean ppm fluoride in adult worker honeybees from smelter, indus-
trial, and urban areas.
166
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TABLE 5.4. FLUORIDE CONTENT OF BEES OVER-WINTERED IN THE COLSTRIP AREA COM-
PARED TO BEES TRANSPORTED TO CALIFORNIA AND BACK INTO THE AREA.
Site
BNE-4
BNE-3
BNE-2
BSE-2
BS-1
x total
SD
SE
N
PPM
x F
Over-wintered
8.9
8. 1
10.2
6.8
9.0
8.4
2.73
.68
16
x F
Transported
8.5
10.0
8.2
7.4
6.5
8.3
2.24
.60
14
have all the characteristics of an excellent source of honey and pollen. By
August 1976, the bees at this location contained 74 to 108 ppm fluoride, the
highest fluoride level of any of the apiaries in the study area.
Because we are interested in the total amount of pollutants in or on a bee,
we usually do not wash the samples prior to analyses. To test whether fluoride
occurred inside or on the outside of the bodies of the bees, combined samples
were taken from 18 sites and split before drying. One sample from each pair was
dried and ground as per normal procedure; the other was rinsed with distilled
and deionized water until the water ran clear. A t-test for the difference in
means for each pair (tr-jg-i = 1.53) was not significant (p >.05), except in a
sample from BNE-10 in wrncn the fluoride content was decreased from 60.7 to 27.6
ppm by rinsing. The average fluoride in bees from each of the paired samples
from the other 17 sites did not exceed 12 ppm. Low concentrations of fluoride
did not easily wash off or out of the bee tissue, whereas at higher levels at
least part of the fluoride did so.
Table 5.5 presents the distribution of fluoride in the three divisions of
the body. Expressed as concentration of fluoride in the body or per body
region, there was no significant difference (p >.05) in the amount of fluoride
in body regions of the bees demonstrating elevated fluoride levels. Bees with
relatively low fluoride content contained the least fluoride (ppm) in the ab-
domen region. However, the absolute quantity of fluoride (in ug/ body region)
in all bees was the greatest in the thorax, which had the greatest mass.
167
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TABLE 5.5. FLUORIDE CONTENT AND WEIGHTS OF DIFFERENT BODY DIVISIONS (HEAD,
THORAX WITH LEGS AND WINGS, ABDOMEN) OF ADULT WORKER HONEYBEES.
(Each value is the mean of four sub-samples from
a combined sample taken from 8-12 hives.)
F Concentration
Body Division (ppm) Mean Dry Weight Site
N x SD (mg)
Whole Bees
Head
Thorax
Abdomen
Whole Bees
Head
Thorax
Abdomen
196
527
527
527
204
898
898
898
97.
97.
97.
97.
1
1
1
1.
0.
2.
7.
9
7
1
6
4
3
8
6
1.
1.
1.
0.
0.
2.
0.
0.
41
3
03
5
6
2
5
72
26.29
1
26
4
14
7
5.71
6.52
8.39
.49
.29
.02
.19
NE-10
NE-10
NE-1
NE-1
S-4
S-4
S-4
S-4
0
0
During 1975 bees were collected at different dates throughout the season
and analyzed for fluorides. The accumulation of fluoride recorded according to
date of collection was not statistically significant (Table 5.6).
Recent (within 24 hours) dead or dying bees collected in August 1975 from
traps or from the front of hives contained 8.0 ppm F , SD = 2.24, SE = .60, N =
14. Bees, from these same hives, that had been dead several days or even weeks
and had decomposed or dried from exposure to the weather contained 4 ppm F , SD =
3.51, SE = 1.42, N = 6, indicating that fluoride was fairly stable even in
decomposed or dried tissue.
The fluoride content of bees, pollen, and vegetation collected near Fort
Howes in the autumn of 1975 indicated that fluoride may accumulate or concen-
trate when transferred from plants to pollen to bees (Table 5.7).
*
In August of 1975 and August of 1976, 114 samples of pollen and bees from
the same hives were collected. The fluoride level in bees was four (1975) to six
(1976) times greater than the fluoride in pollen. European investigators have
reported that contaminants are brought into the hive primarily with the pollen
(Debackere, 1973; Toshkov et al_. , 1974). To test whether the fluoride in bees
correlated with the fluoride level in the pollen brought into their hives,
Pearson's product-moment correlation coefficient was utilized. The results are
given in Appendix G, Tables G-l, G-2, and G-3. However, fluoride in bees did not
correlate with fluoride in pollen, and the level of fluoride in pollen was the
same for both years.
168
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TABLE 5.6. PPM FLUORIDE IN BEES COLLECTED ON DIFFERENT DATES IN 1974 AND
1975; SAMPLES FROM EACH SITE COMBINED AT TIME OF COLLECTION.
October
1974
x 7.5
SD 3.09
SE .63
N 24
No. Sites 12
May
1975
9.3
2.3
•77
10
9
June
1975
8.7
1.08
.44
22
17
July-August
1975
7.1
1.77
.42
18
13
August-September
1975
9.5
2.38
.56
18
18
TABLE 5.7. PPM FLUORIDE CONCENTRATIONS IN HONEYBEES, POLLEN, AND BLOSSOMS.
Honeybees Pollen Sweet Clover Alfalfa Snakeweed All Vegetation
X
SD
SE
9.
1.
1.
13
74
01
1.3
.34
.17
.7
.56
.23
.4
.44
.18
1.3
.14
.10
.8
.61
.16
Efforts were concentrated on late-season collections of bees and bee
materials because during late August or early September: (1) Bee colonies were
at their peak in size; (2) most of the honey supplies had been gathered and
stored; (3) pollen supplies were at a maximum as egg laying and brood rearing
declined in response to diminishing supplies of nectar; (4) the alfalfa seed
crop had been pollinated; (5) colonies had had a sufficient period of exposure
to any environmental contaminants in the region, and (6) there had been several
turnovers in bee populations and supplies so that any contaminants brought in
from California were likely to be eliminated or greatly diluted. Because the
efforts were concentrated on ZAPS in 1976, all of the outlying yards did not get
sampled, as had been done in 1975. The results of the fluoride analyses for bees
and pollen collected in August of 1975 and 1976 are presented in Appendix H.
Collecting bees before mid-June proved to be impractical because some
colonies had been returned from California, others had not, and those that were
in the area were often weak, reducing the number of bees which could be sampled
without risking harm to the colony. In May of 1975, there were only ten colonies
in the Colstrip area from which to take samples, but the average fluoride level
of 9.2 ppm was consistent with fluoride levels at these same sites later in the
season (Table 5.6). The most distant site, near Biddle, Montana, had higher
fluoride in the bees in the spring than later in the season (30.2 ppm, May; 7.9
ppm, August). Needle pathologies typical of pollution damage were observed and
169
-------
increased sulfurs and fluorides were measured in pines approximately .6 km from
this yard, indicating a nearby pollution source. The Biddle site is less than 8
km from oil fields which have occasional burn-offs that may impact the nearby
apiary and pine locations. Also, bees are medicated in the spring, and each bee
colony consists mainly of over-wintered or older bees, all of which could
account for higher levels of fluoride.
The high fluoride level in bees at BNE-10 was surprising, and the fluoride
in the bees did not correlate with the level of fluoride in pollen. There was no
known industrial source of fluoride nearby, and other possible sources such as
automobile emissions or the adjacent railroad were encountered at other
locations, some of which had very low fluoride baselines. The water supply of
the bees was hypothesized as a potential source of fluoride. The Yellowstone
River, a slough approximately 20 m away, and a stock tank fed by a constantly
flowing artesian spring (the source of the water in the slough) were readily
available water sources. The fluoride content of these waters in May of 1976
were, respectively, 0.4 ppm, 11.8 ppm, and 7.0 ppm. Well and surface waters in
the vicinity of Colstrip (1 to 20 km) contained 0.0 to 0.6 ppm, with a mean of
approximately 0.2 ppm (unpublished, Bureau of Mines and Geology, personal
communication, R. Hedges). In August 1976, the waters were sampled where bees
were seen drinking at all of the apiaries near Colstrip. Most of the bees
obtained their water from Rosebud Creek which had a fluoride level of 0.6 ppm at
the majority of the sites. Mean fluoride in bees versus mean fluoride in water
for each site had a correlation coefficient of 0.99658 (Figure 5.9), which is
significant (p < .001). However, this high correlation may result from grouping
data from a site with a high level of fluoride in bees and water with data from
all other sites at the base of the regression line, with no intermediate values.
Bees from an apiary 2.5 km from the Corette power plant at Billings averaged 39
ppm F , and water from the river near that site averages about 1 ppm F , accord-
ing to Montana State Department of Health records. Water appears to affect
fluoride in bees, and we shall attempt to refine this observation in 1977 by
obtaining bees and water from areas where the level of fluoride in bees falls
between the extremes of these specific cases.
The bees at BNE-10, which is near the town of Rosebud, Montana, averaged 7
to 12 times higher fluoride than any other site in the study area; while the
water had 16 to 20 times the fluoride as water at other sites. These levels of
fluoride in these bees were indicative of a pollution source and were just
slightly less than the level of soluble fluoride forms reported to be toxic to
bees. The level of fluoride in bees at all other sites was indicative of a clean
area (Lillie, 1970) and as such served as a baseline value. Statistically, the
results of Bartlett's test of homogeneity of variances and appropriate t-tests
for all collection dates indicated that the Rosebud bees were significantly
different (tr-,-,-,-, = 9.91, p < .001) in terms of fluoride content from bees from
all other sampiejsites in southeastern Montana. The mean value of fluoride in
bees at Rosebud fell outside of two standard deviations of the mean for all
sites. Therefore, the fluoride data for the bees, the existence of a fluoride
source (water), and literature citations indicated that this site was not
representative of a baseline level and as such was not directly comparable to
the other sites. For these reasons, the data from Rosebud has been excluded in
treatment of the data from the study area sites.
170
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oo
LU
LU
CQ
D:
o
50 -
40 -
30 -
20 -,
10
r = 0.997
y = 5.692 + 4.044x
I I
6 8
FLUORIDE IN WATER
I
10
1
12
Figure 5.9. Fluoride in adult worker honeybees vs. fluoride in water.
171
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Tables I.I, 1.2, 1.3, and 1.4 in Appendix I present the analysis of
variance for fluoride in bees collected in August of 1975, just before the first
power generating unit went on-line, and for bees collected in August of 1976
(both Colstrip units have been operating at approximately one-third capacity).
The data demonstrates significant increases of fluoride in bees at sites S-5,
SE-1, NE-2, NE-3, and NE-4, southeast and northeast of Colstrip, while fluoride
in bees at other sites remained about constant or decreased (Figure 5.10). Wind
data was supplied by the Montana State Department of Health for the period of
May through September of 1976, and obtained from a 300-foot tower atop a 200-
foot hill, 1,400 m north-northeast of the power plants. The most prevalent
winds blow to the east-southeast, the west-northwest, and the east-northeast,
respectively. Note in Figure 5.11 that the wind blows least in the direction of
those apiaries showing little or no fluoride increase. There are no apiaries in
the direction of the second most prevalent winds, i.e., to the west-northwest.
Wind patterns and fluoride increases are depicted in Figure 5.11.
Bees were collected during two periods in 1976, late July and early August.
The fan motor of the electric sampler burned out at the first site in July. The
motor was not immediately repairable, so bees were collected by removing the
outside, uppermost two frames from each hive and sweeping the bees into plastic
bags. Two weeks later these same hives and apiaries were collected using the
repaired suction apparatus. To check whether the altered procedure (sweeping)
affected analyses results, hives were sampled at each yard using both methods.
The results are given in Table 5.8. Bees inside the hives had 50 percent less
fluoride than bees collected at the entrance to the hive. Thus, the two sam-
pling procedures gave significantly different results, although the results of
the sweeping were consistent for both sample periods. This prohibited the use
of the July 1976 data for meaningful comparisons with any collection taken with
the vacuum apparatus.
TABLE 5.8. COMPARISON OF MEAN FLUORIDE IN ADULT WORKER HONEYBEES COLLECTED BY
SWEEPING BEES FROM THE WAX COMBS AND BY VACUUM SAMPLING AT THE
ENTRANCE TO THE HIVES.
August, 1976 September, 1976 September, 1976
Sweep Sample Sweep Sample Vacuumed Sample
(ppm F ) (ppm F ) (ppm F~)
x = 5.
SD = 3.
SE = 0.
N = 61
83
25
42
x =
SD =
SE =
N =
5.
3.
1.
12
87
88
12
x
SD
SE
N
= 12.
= 4
= 0.
= 45
05 *
48
67
Honeybee colonies have a division of labor, depending for the most part on
the maturation of the worker bees. The eldest bees in a colony are the foraging
bees, the bees most likely to be actively flying in and out of the entrance of
the hive. Bees building comb, receiving nectar from foragers, and removing
172
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Q_
s:
co
SW-1
SW-2
SW-3
S-l
S-4
S-5
SE-1
SE-2
NE-2
NE-3
NE-4
N-2
SE-12
1975 (
1976 (
12
PPM FLUORIDE
16
20
24
Figure 5.10. Mean fluoride in adult worker honeybees, each value is the mean
of four independent samples with 95% confidence interval.
173
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YELLOWSTONE RIVER
Figure 5.11. 1976:75 mean fluoride ratios for honeybees near Colstrip,
Montana, and wind rose.
174
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debris, cell caps, and materials from the bottom of cells from the hive, tend to
be of intermediate age. Differences in the average activities of individuals of
a given age may occur among colonies or within the same colony at different
times (Michener, 1974).
In 1975 and 1976, honey samples were collected from the same hives from
which pollen and bees were obtained. Honey analyses are in process, but we are
not satisfied with the efficacy of the analyses procedures. Honey is extremely
hygroscopic and as such requires special handling to remove the water. This has
been accomplished via acid digestion and subsequent drying. Hillmann (1972)
utilized a relatively complex procedure to analyze honey with the Leco furnace,
which required continual readjustments of oxygen flow and of rheostat settings,
and which was less than satisfactory because of occasional explosions which sent
glass flying around the laboratory (personal communication, R. Hillmann).
Currently we have been searching the literature and experimenting in an effort
to improve the honey analyses.
Sulfur analyses for the three years yielded values of 4,392 ± 286 (SE =
82.3), 4,000 ± 436 (SE = 251), 4,881 ± 497 (SE = 143.64) ppm, respectively.
These differences were not significant (p >.05). Hillmann (1972) failed to
detect significant differences between bees fumigated for periods of nine and 14
weeks with 1, 3, and 5 ppm S02 and those from the controls. His explanation that
this was due to the comparatively higher sulfur content of the protein in bee
tissues, which masked any additional amounts attributable to the fumigation, may
be correct. Honey, which is primarily carbohydrate in composition, may provide
a means to monitor sulfur accumulation in bee systems without the complication
incurred when utilizing bee tissue. This is one of the main reasons for
analyzing honey, especially since a study by Tong ^t a!. (1975) indicates that
honey does accrue contaminants such as sulfur from the environment. Another
approach is to analyze bee tissue for specific sulfur compounds such as sulfate
and sulfite. Gunnison (1970) was able to detect significant increases (twofold)
of sulfur in the hemolymph of bees after fumigation with 8.2 ppm S02 for three
weeks. However, Gunnison eventually decided to use rabbits for this work
because of insufficient quantities of hemolymph, and his procedures were
extremely difficult and time consuming.
A third possible approach to investigating the effect of S02 on honeybees
is to analyze for acetylcholinesterase (Ache) activity, which reportedly is
inhibited or decreased by sulfur dioxide. Dr. R. J. Barker, USDA Bee Research
Laboratory, Tucson, Arizona, conducted Ache enzyme analyses on heads or brains
from bees (both foraging and dying bees) from the ZAPS and from one of the
southeastern Montana control sites (Table 5.9). He found that the enzyme activ-
ity in bees from the ZAPS and the control was as high or higher than the enzyme
activity of control bees from the Tucson area, and considerably higher than the
enzyme activity rates of bees poisoned by organophosphate pesticides from his
own research project. The failure to detect a change in the Ache activity of
bees at the ZAPS may be due to the fact that many of the bees in each colony were
not exposed to the sulfur dioxide gas because the hives were placed 75 m north of
the plots to avoid conflicts with the investigators working on the plots. In
addition, determinations of Ache activity levels in the thoracic ganglion may be
a more reliable and sensitive indicator than Ache activity levels in the brain
(personal communication, W. Stephen, Oregon State University). We do not intend
175
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TABLE 5.9. COLORIMETRIC DETERMINATION1 OF ACETYLCHOLINESTERASE ACTIVITY IN
HEADS OF HONEYBEES FROM ZAPS AND COLSTRIP PLOTS.
Sample2
A 900
A 901
A 902
A 903
A 904
A 905
A 906
A 907
A
B
Number of
Heads
5
5
5
5
5
5
5
5
5
5
Weight of
Heads (g)
0.0428
0.0408
0.0477
0.0463
0.0475
0.0396
0.0389
0.0282
0.071
0.0323
Buffer (ml)
6.4
6.1
7.2
7.0
7.1
5.9
5.8
4.2
2.6
4.8
Rate
5.6
5.1
5.2
5.3
5.6
6.6
5.8
7.1
3.7
1.5
Rate/ml
28.0
25.5
26.0
26.5
28.0
33.0
29.0
35.5
18.5
7.5
Rate/head3
35.84
31.11
37.44
37.10
39.76
38.94
33.64
29.82
9.62
7.2
1R. J. Barker, USDA-ARS, Tucson, Arizona.
2Samples A 900 - A 907 from Eastern Montana, Samples A and B from Phoenix.
Phoenix bees were poisoned by pesticides.
3A11 rates for bees from ZAPS are normal.
to pursue the feasibility of Ache as an indicator of sulfur dioxide effects
and/or toxicity until it is ascertained (through methods such as the use of
sulfation plates or via sulfur in honey) that the bees are being exposed to the
sulfur dioxide either near Col strip or at the ZAPS (see ZAPS report).
Bees were collected for pesticide analysis primarily from the area along
the Rosebud Creek drainage near Colstrip, although bees were also obtained
during 1975 from apiaries near the communities of Rosebud, Broadus, Ashland,
Fort Howes, and Biddle. The localities sampled in 1975 were distributed over a
broader area than those sampled in 1974 to provide a means of detecting and
monitoring pesticides throughout the study region. Bees from the following
sites were analyzed for pesticides:
176
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1975 1974
BNE-2 BNE-3
BNE-3 BNE-4
BNE-4 BNE-5
BNE-5 BSE-1
BNE-10 BSE-2
BSE-1 BS-1
BSE-2 BS-4
BSE-9 BS-5
BSE-10 BSW-2
BSE-12 BE-1
BS-4 BN-1
BS-5
BS-9
BSW-2
All of the 1975 samples and seven of the 1974 samples contained traces
(0.01 to 0.03 ppm) of the DDT/DDD/DDE chlorinated hydrocarbon complex. Thomas
reports that this is not an unusual finding in environmental samples. There was
no gross contamination in any of the samples. No residues of carbamate or
organophosphate pesticides were found. Because pesticide analyses require
large amounts of bee tissue, it was decided to discontinue routine pesticide
sampling and restrict samples utilized for pesticides to those apiaries at which
symptoms of bee poisoning were observed.
The results of the analyses of honeybees and pollen for the last three
years have confirmed the original hypothesis that honeybees are useful in the
detection and collection of toxic materials (fluoride) from the environment.
Significant (up to two times) increases were detected in the mean fluoride in
bees at several apiaries located 11 to 15 km southeast and northeast of Colstrip
over the average fluoride levels for the two years prior to the operation of the
Colstrip power plants. Fluoride levels in water apparently play a role in the
baseline levels of fluoride; fluoride in pollen may play a less important role,
although Maurizio (1956) thought that bees poisoned by industrial effluents in
Switzerland were obtaining fluoride via the pollen and not via water. Toshkov
et al. (1974) found that bees transported poisonous substances (e.g., copper,
zinc, and phosphorous) from the environment into the hives mainly by means of
pollen and, to a much lesser degree, by means of nectar. They did not report
analyses of water supplies. It is probable that the routes by which fluoride
enters a bee colony may be varied and rely on the forms (e.g. , gas, particulate,
liquid) of the contaminant. Debackere's review (1972) of air pollution and
apiculture makes the following observations: (1) The gaseous forms of fluoride,
such as hydrogen fluoride and silicates of fluoride, are more dangerous to bees
than the powdery compounds; (2) there is considerable disagreement over the
toxicity of industrial fluorides to bees; (3) the diet of bees can increase
manifestly their tolerance for fluorine, and (4) fluorine compounds cannot
penetrate the thoracic chitin and are toxic primarily by action through the gut;
for this reason, solubilized forms are said to be more toxic. He describes
fluorine poisoning as a long-term mortality which can lead to the complete loss
of the colony, but this loss occurs gradually, especially during the summer.
The action of fluorine is that of a stomach poison; presumably fluorine can be
177
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fixed by lime particles in the epithelial cells of the digestive tract, and this
exerts a protective function. The latter action would explain the increased
tolerance conveyed by albumen feeding and would also explain the increased
susceptibility of bees infected with Nosema disease, which affects the
epithelial cells of the intestine. The long-term course of fluorine poisoning
of bees could account for the lack of vigor, weak colonies, and poor production
by the bees at Rosebud. The levels of fluoride in these bees approached that
considered to be definitely toxic (LD50) if the fluoride is in a soluble form,
which is the form of fluoride that is measured in the water tested with the Orion
specific ion electrode.
The mean fluoride level in bees from Billings, an area known to be sub-
jected to fluoride pollution, was 30 ppm, which is approximately the level that
has been reported indicative of a permanent or intermittent source of fluoride
contamination (Guilhon, 1962). This is about five times higher than the levels
found in Colstrip prior to the operation of the Colstrip units, and is about
three times as high as the average fluoride in bees near Colstrip in 1976.
Sulfur accumulation in bees does not appear to be particularly useful in
discerning accumulative effects, probably because of a masking effect by the
high sulfur content of the protein in bee tissues. The results of the fluoride
studies have encouraged us to examine the accumulation of other major and trace
elements (such as arsenic, cadmium, and lead) which are known to be emitted
during the combustion of coal.
Honeybees have been advocated previously (Toshkov e_t al. , 1974) as bio-
indicators of impurities in the environment. Toshkov reports that honeybees may
forage as far as 6 to 8 km from the hive or over an area of 10,000 acres. Our
data shows significant increases in fluoride in bees from those apiaries located
along major wind pathways from Colstrip. If this fluoride is coming from the
Colstrip units, then fluorides are being carried at least 7 to 10 km from the
power generators, even considering the extent of the bees' flight range. Based
on the .results of our current studies, honeybees appear to be excellent
indicators of fluorides in the environment. In fact, they are sensitive to
changes of a few parts per million and may be accumulators and magnifiers of
fluoride, at least as evidenced by the magnitude of increase in bees while the
levels of fluoride in water and pollen remained about constant.
If the available resources of bees are not strained in an apiary by
conducting analyses which require large numbers of specimens, such as pesticides
analysis, one should be able to take a greater number of individual samples and
improve the usefulness of the data both in terms of improving the sample size
and in terms of obtaining a better estimate of variability. One of the possible
effects of increased fluoride in the system may be an increase in the variation
within an apiary, since bees from each hive tend to forage in a specific area.
Thus, the individuals from any one hive could be exposed to a different level of
fluoride in the environment. At baseline levels, this variability may be less
as each colony is exposed to a relatively constant or stable level of the
contaminant. It is our intention to increase the number of individual samples
and to reinstate collections at different times of the growing season.
178
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SUMMARY AND CONCLUSIONS
Insect and plant studies have been carried out since August 1975 to
establish baseline data on populations of selected species of indigenous insects
and plants. The major hypothesis was that one could develop methods to predict
the bioenvironmental impacts of coal-fired power plants before damage occurred.
These methods were based on the use of indicator species of insects and plants
as early warning systems and as continuous monitors of atmospheric pollutants.
This approach was unique because it allowed baseline studies in an area that was
one of the cleanest, most unpolluted areas in the contiguous United States
before the Colstrip power plants began operations (EPA, 1976; DHES, 1976; DNRC,
1975; WES, 1973). These studies and those of numerous other investigators, both
under this EPA-sponsored project or other federal, state, and private
sponsorship, have provided extensive baseline data of characteristics of this
clean or pristine area. Almost all published studies concerning air pollution
impacts on terrestrial ecosystems have either dealt with post-impact studies,
which lacked pre-impact baseline information, or with controlled fumigations in
laboratory chambers.
We believed (and believe) that biological indicators would show the actual
responses through time of organisms or populations to individual and synergistic
effects of combined pollutants or stresses. However, the bioindicators had to
be sensitive to perturbations, easy to measure and monitor, and reliable
predictors of bionomic and economic impacts. To test our hypothesis and to meet
these criteria, we proposed and completed these objectives: (1) Selection and
establishment of permanent study sites distributed to maximize gradient
exposures to emissions from the Colstrip complex; (2) compilation of inventories
of insect populations, infestations, and damage to indigenous plant species at
the study sites to select specific groups for use as bioindicators; (3) analyses
and selection of insect populations with a diversified but understandable
interrelationship with indigenous plant species in order to ascertain effects of
air pollutants on symbiotic associations; (4) selection and testing of disease-
and injury-causing insect species for more intensive study; (5) selection and
testing of beneficial insects for more intensive study, and (6) chemical
analyses of selected plants and insects to establish baseline levels and to
discern any accumulation of materials (mainly fluorides and sulfurs) released by
the combustion of coal.
For the entomological investigations, 17 sites in ponderosa pine-skunkbush
ecosystems were established on exposed ridgetops facing Colstrip, and 25
apiaries on the river and creek drainages along the interface between the
grassland and the cropland ecosystems. The investigators from the Colorado
National Resources Ecology Laboratory inventoried and quantified population
characteristics of arthropods in the grassland areas. Therefore, we decided to
compile information concerning the major or dominant insect populations on
ponderosa pines, a species which research has shown (Gordon et al_. , 1976; Gordon
et a!., 1977) to be one of the most sensitive plant species to the phytotoxic
gases" released by coal-burning facilities. This pine is dominant throughout
much of southeastern Montana and is the sole commercial timber resource. In
addition, two points were of critical importance: (1) Many reports indicated
that conifers weakened or injured by pollutants are more likely to incur attack
from insects that prey and reproduce on weakened trees (Haegle, 1973), and (2)
179
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phytophagous insects have long-term interactions with fundamental ecosystem
processes such as primary production and nutrient cycling (Mattson and Addy,
1975). We quantified pathologies of trees and foliage and established data
banks concerning baseline levels of sulfur, fluorides, and trace elements. The
insect surveys identified prevalent species of both pest and beneficial insects
associated with the pine, evaluated the status of insect pests and their
immediate potential for outbreak or epidemic, determined which insect species
most likely were responsible for insect-induced pathologies, and provided
histopathological demonstrations of insect damage to pine foliage. The latter
were of particular value when juxtaposed with the histopathological micro-slide
series of fungal, pollutant, and abiotic (e.g., frost, drought) damage
syndromes. Histopathological descriptions allowed us to "fingerprint" damage
from the initial damage through each stage as it first becomes visible and then
fully manifest. We already have a "fingerprint" file for many types of injury
from phytotoxic gases and from fungi. We quantified foliage damage by insects
and other sources, rather than attempting to quantify the absolute size of pest
insect populations. Ultimately, it was the amount of injury that was the most
meaningful measure of the effect of an insect population on its host. Based on
pre-operational studies, the southeastern Montana area was considered to be
exceptionally free of serious insect problems, as evidenced both by insect and
disease reports and by our own observations.
One insect, identified as a Matsucoccus scale, is of the highest interest
because it resides at the base of the needles under the fascicular sheath, is
often seen near areas of basal necrosis or basal burn to the needles, as is
typical of acid rain damage, and is reported to increase in numbers in areas of
pollution stress (Siewniak, 1971). The life system of this insect was
concentrated on, including its interactions with ponderosa pine, and the
pathological symptoms of its feeding and attack. This is an ongoing objective;
its relationships with ponderosa pines are still not fully understood. The
results of this work should make a significant contribution not only towards
understanding the role air pollution plays in the population dynamics of this
organism but also to an understanding of the basic biology of this insect, which
is seldom studied by researchers in the United States and Canada, although some
species have caused extensive damage in the United States (Keen, 1952).
The post-operational investigations did not demonstrate any significant
increase in insect pests or insect damage at any of the sites, but our chemical
analyses and foliage pathology determinations demonstrated significant
increases in sulfur and fluoride in plant foliage at two sites and significant
increases in needle pathologies of 14- to 26-month-old needles collected from
one of these sites (see section by Gordon et a 1., this report).
Honeybees were utilized near Col strip as accumulators and indicators of the
extent and magnitude of the phytotoxic and zootoxic effluents from the Colstrip
power units. The 1974, 1975, and 1976 honeybee collections demonstrated
significant differences among sites in the fluoride content of bees. However,
the pre-operational levels of fluoride in bees prior to the operation of the
Colstrip units averaged 8.5 ppm for 22 sites in 1975 and 7.4 ppm for 11 sites in
1974. One site was shown to have a source of fluoride contamination (10 to 12
ppm) in the bees' water supply, and the bees at that site averaged 96 ppm for all
years. Honeybees collected at apiaries approximately 15 to 20 km northeast and
180
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southeast of Colstrip in September 1976 (12 months after Colstrip Unit 1 went
on-line), demonstrated a significant (p £.001) fluoride increase over the levels
found at these sites during the previous two years. Increases as high as
twofold occurred at these sites, which are in the path of frequent winds from
Colstrip, while slight decreases in fluoride were demonstrated at sites north
and south of Colstrip in the path of the least frequent winds. Both 1975 and
1976 pollen samples averaged 1.9 ppm for all sites. Fluoride in water at the
sites, which hypothetically could be responsible for significant increases of
fluoride in bees, was practically constant at 0.6 ppm. Since honeybees may
forage up to 6 to 8 km (Toshkov et a]_. , 1974) from their hives, this would mean
that if the bees are accumulating fluoride emitted by the Colstrip units, these
effluents are being carried at least 7 to 10 km southeast and northeast of
Colstrip.
The studies indicated that honeybees may be extremely sensitive and useful
indicators of fluorides in the environment, and this prompted us to design a
study for the 1977 field season investigating the presence of other major and
trace elements. Honeybees do not appear to be particularly useful for the
detection of sulfurs in the environment because the natural levels of sulfur in
bee tissues are so high that they apparently mask any measurable increases. The
results of analyses of honeybees for the presence of pesticide residues in 1974
and 1975 revealed only low levels (0.01 to 0.03 ppm) of chlorinated hydrocarbons
in any of the samples, a result that is not unusual in these types of samples.
The results of the baseline studies of the accumulation of phytotoxic
substances of anthropogenic origin in vegetation, honeybees, and pollen; the
field surveys of pest insect populations and infestations on ponderosa pines and
associated measurements of foliage pathologies; and the field studies of insect-
plant life systems (namely M. secretus on ponderosa pine) reaffirm the belief
that one should be able to follow changes in the baseline characteristics of the
pine and grassland ecosystems of southeastern Montana as they are exposed to
emissions from the Colstrip power plants.
Furthermore, it is believed that these entomological investigations
contribute to both basic and applied fields of air pollution research about
changes in systems of insects and insect-plant interactions and about the
pathways of air pollutants through an insect system, honeybees in this case.
The information on the responses of pollinators to effluents from coal-fired
generating plants should be invaluable to other EPA studies on plant diversity,
plant productivity, and plant community changes, while the work on the ponderosa
pine systems adds this prominent ecosystem to the grassland ecosystem studies of
the overall project.
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II Fecundity, Hatchability, and Fertility. Atm. Environ.
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12, U.S. Forest Service, Missoula, Montana.
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183
-------
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«
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184
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185
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APPENDIX A
LITERATURE AND REPORTS RELEVANT TO INSECT-TREE-AIR POLLUTION INTERACTION
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the vicinity of Oakland, Maryland and Mount Storm, West Virginia.
Report submitted to U.S. Dept. H.E.W., N.A.P.C.A., Durham, N.C.
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trees in the area of Mt. Storm, West Virginia, and of caged experiments
(Modified Moch's Postulates). Unpublished Report. North Carolina
State University, Raleigh, N.C. 38 pp.
Baker, J. R. 1973. Final report on caged experiment involving Adelgids as
a possible cause of needle shortening of Scotch pine. Unpublished
Report. North Carolina State University, Raleigh, N.C. 15 pp.
Bb'sener, R. 1969. Zum Vorkommen ridenbrutender Schadinsecten in rauch-
geschadigten Kiefern-und Fichtenbestaenden. Arch. Forstwes. Bd. 18:1021-
26.
Bromenshenk, J. J. 1975. Biological Impact of Air Pollution on Insects.
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Meeting Twenty-Sixth Annual Western Forest Insects Work Conference
Twenty-second Annual Western International Forest Disease Work Confer-
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Forestry Service, Edmonton, Alberta. 86-96.
Bromenshenk, J. J. 1976. Investigations of the Effects of Coal-Fired Power
Plant Emissions Upon Insects, Report of Progress. In: The Bioenviron-
mental Impact of a Coal-Fired Power Plant, Second Interim Report,
Colstrip, Montana - June, 1975. R. A. Lewis, N. R. Glass, A. S. Lefohn,
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186
-------
Bromenshenk, J. J. and C. E. Carlson. 1976. Impact on Insect Pollinators.
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Twenty-second Annual Western International Forest Disease Work Confer-
ence. Intermountain Forest and Range Experiment Station U.S.D.A.
Forest Service, Ogden, Utah and Northern Forest Research Center, Canadian
Forestry Service, Edmonton, Alberta. 86-96.
Carlson, C. E. and J. E. Dewey. 1971. Environmental pollution by fluorides
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57PP.
Carlson, C. E. and W. P. Hammer. 1974. Impact of Fluorides and Insects on
Radial Growth of Lodgepole Pine near an aluminum smelter in northwestern
Montana. A preliminary inquiry. U.S.D.A. Forest Service, Northern
Region, Div. S&PF, Missoula, Montana. Rep. No. 74-25.
Carlson, C. E. , M. D. McGregor, and N. M. Davis. 1974. Sulfur damage to
Douglas-fir near a pulp and paper mill in western Montana. U.S.D.A.
Forest Service, Northern Region, Div. S&PF, Missoula, Mt. , Rpt. No. 74-
13, 41 pp.
Carlson, C. E. , W. E. Bousfield, and M. D. McGregor. 1974. The relationship
of an insect infestation on lodgepole pine to fluorides emitted from a
nearby aluminum plant in Montana. U.S.D.A. Forest Service, Northern
Region, Div. S&PF, Missoula, Mt., Rpt. 74-14, 21 pp.
Ciesla, W. M. 1975. The role of atmospheric pollutants in predisposing
trees to insect attack. In: Air pollution and insects. Carlson,
C. E. (Moderator). Proceedings Joint Meeting Twenty-Sixth Annual
Western Forest Insect Work Conference Twenty-second Annual Western
International Forest Disease Work Conference. Intermountain Forest and
Range Experiment Station U.S.D.A. Forest Service, Ogden, Utah and
Northern Forest Research Center, Canadian Forestry Service, Edmonton,
Alberta. 86-96.
Cobb, F. W. Jr., D. L. Wood, R. W. Stark, and P. R. Miller. 1968a II.
Effect of injury upon physical properties of oleoresin, moisture
content and phloem thickness J_n Photochemical oxidant injury and bark
beetle (Coleoptera:Scolytidae) infestation of ponderosa pine. Hil-
gardia. 39(6):127-134.
Cobb, F- W. Jr., D. L. Wood, R. W. Stark, and J. R. Parmeter, Jr. 1968b
IV. Theory on the relationships between oxidant injury and bark beetle
infestation j_n Photochemical oxidant injury and bark beetle (Coleoptera:
Scolytidae) infestation on ponderosa pine. Hilgardia. 39(6):141-157.
187
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Compton, 0. D.
condition
Agri. Exp.
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of ponderosa pine trees in the Dalles area. Misc. paper 120,
Sta. , Oregon State University, Corvallis, OR. 6p.
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source in western Montana. Environ. Ent. 2(2):179-182.
Donaubauer, E. 1966. Secondary Damages of Forests Caused by Industrial
Exhaust Fumes. (Durch Industrie-abgase bedingte Sekundaerschaeden am
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277-284.
Edmunds, G. F. , Jr. and R. K. Allen. 1958. Comparison of black pine leaf
scale population-density on normal ponderosa pine and those weakened by
other agents. Int. Congr. Entomol. Proc. (Montreal, 10th., 1956)
4:391-392.
Edmunds, George F
2(5):765-777.
1973. Ecology of black pineleaf scale. Env. Entomol
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of Ent., Forest Insect Investigations. Typewritten report on file at
R-l , Missoula, Mt.
Farrier, M. H. 1972. Report on insects and mites in relation to the long-
short needle syndrome of Scotch pines and their abundance in Christmas
tree plantations in western Maryland and northern West Virginia.
Unpublished Report. North Carolina State University, Raleigh, N.C. 57
pp.
Fischer, G. W. 1950. Second progress report Spokane County ponderosa pine
blight investigation. U.S.D.A. Forest Service. Unpublished Report. 25
P-
Gordon, C. C. 1976. A preliminary study of fluoride concentrations in
vegetation samples collected September 8 and 9, 1976 in and 'around the
town of Kitimat, B.C., Canada.
Gordon, C. C. , C. E. Carlson, and P. C. Tourangeau. 1976. A cooperative
evaluation of potential air pollution injury and damage to coniferous
habitats on National Forest lands near Colstrip, Montana. I. Interim
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188
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Hay, C. J. 1975. Arthropod Stress. In: Air Pollution and Metropolitan
Woody Vegetation. W. H. Smith and L. S. Dochinger (Eds.) Yale Uni-
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Heagle, A. S. 1973. Interactions between air pollutants and plant para-
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pollutants. Phytopathology. 58:1098-1101.
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and Mesophyll-feeding leafhoppers. For. Sci. 15(2):154-157.
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273. 240 pp.
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to timber damage in the smelter fume area near Northport, Wn. U.S.D.A.
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file at R-l. Missoula, Mt.
Kudela, M. and E. Novakova. 1962. Lesnf skudc: a skudy zveri v lesich
Poskozovanych Kourem. Lesnictvi. 6:493-502.
Laurent, T. H. and B. Baker. 1975. Associations of high defoliating insect
populations and high foliar chemical levels near two pulpmills in
southeast Alaska. In: Air pollution and insects. Carlson, C. E.
(Moderator) Proceedings Joint Meeting Twenty-Sixth Annual Western
Forest Insect Work Conference Twenty-second Annual Western International
Forest Disease Work Conference. Intermountain Forest and Range Experi-
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Research Center, Canadian Forestry Service, Edmonton, Alberta. 86-96.
Linzon, S. N. 1958. The influence of smelter fumes on the growth of white
pine in the Sudbury region. Canadian Dept. Agr. Publ., Ontario Depart-
ment of Lands & Forests. 45 pp.
Linzon, S. N. 1966. Damage to eastern white pine by sulfur dioxide,
semimature-tissue needle blight, and ozone. J. Air. Pollut. Cont.
Assoc. 16:140-144.
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Miller, P. R. , F. W. Cobb, Jr., and E. Zavarin. 1968. III. Effect of
injury upon oleoresin composition, phloem carbohydrates and phloem pH
J_n Photochemical oxidant injury and bark beetle (Coleoptera:Scolytidae)
infestation of ponderosa pine. Hilgardia. 39(6):135-140.
Pack, M. R. and B. A. Zamora. 1976. Pre-operational Report. Forest eco-
system monitoring for sulfur dioxide damage in the area surrounding the
Northwest Alloys Magnesium Plant at Addy, Washington. Wash. State
Univ. Publ., Pullman, Washington. 148pp.
Pfeffer, A. 1963. Insektenschadlinge an tannen im bereich der gasexhala-
tionen. Z. Angew. Entomol. 51:203-207.
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Sozialistische Fortwirtschaft (Berlin). 18:299-301, 319.
Saunders, J. L. 1972. Disease and insect pests of Christmas trees. School
for Christmas tree growers. Cornell Univ. Col. Agric. Proc. p. 88-90.
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Scheffer, T. C. and Hedgcock, G. G. 1955. Injury to northwestern Forest
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49 pp.
Schnaider, A. and Z. Sierpinski. 1967. Dangerous condition for some forest
tree species from insects in the industrial region of Silesia. Prace
Instytut Badawczy Lesnictwa (Warsaw) No. 316:113-150.
Sierpinski, Z. 1966. Znaczenie gospodarcze skronsnika tuzinka (Exoteleia
dodecella L. ) na terenach uprzemyskowionych. Sylwan. 110:23-31.
Sierpinski, Z. 1967. Influence of industrial air pollutants on the popu-
lation dynamics of some primary pine pests. Proc. 14th Contr. Int.
Union For. Res. Organ. Vol. 5, sect. 24, Munich. 518-531.
Siewniak, M. 1971. Uszkadzanie sosny pospolitej (Pinus silvestris) przez
czerwca korowinowca (Matsucoccus pini Green 1925; Margarodidae, Coc-
coidea). Sylwan. 115(12):35-41.
Stark, R. W. et ah 1968. I-Incidence of bark beetle infestation in injured
trees i_n Photochemical oxidant injury and bark beetle (toleoptera:
Scolytidae) infestation of ponderosa pine. Hilgardia. 39(6): 121-126.
Templein, E. 1962. On the population dynamics of several pine pests in
smoke-damaged forest stands. Wissenschaftliche Zeitschrift der Tech-
nischer Universitatet Dresden. 11:631-637.
Treshow, M. and M. R. Pack. 1971. Fluoride. In: Recognition of Air
Pollution Injury to Vegetation: A Pictorial Atlas. J. S. Jacobson and
A. C. Hill, eds. Rept. No. 1, TR-7 Agricultural Committee, Air Pollu-
tion Control Assoc.
190
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Weismann, L. and L. Svatarakova. 1973. Influence of sodium fluoride on
behavior of caterpillars Scotia segetum Den. and Schiff. Biologia
(Bratislavia) Czechoslovakia. 28:105-109.
Wentzel. K. F. 1965. Insekten als Immissionsfalgeschadlinge. Naturewis-
senschaft. 52:113.
Williams, W. 1975. Oxidant air pollution damage in California forests.
In: Air Pollution and insects. Carlson, C. E. (Moderator). Proceedings
Joint Meeting Twenty-Sixth Annual Western Forest Insect Work Conference
Twenty-second Annual Western International Forest Disease Work Confer-
ence. Intermountain Forest and Range Experiment Station U.S.D.A.
Forest Service, Ogden, Utah and Northern Forest Research Center, Canadian
Forestry Service, Edmonton, Alberta. 86-96.
Wong, H. R. and J. C. E. Melvin. 1973. Insects associated with trees
damaged by hydrocarbon condensate in the Strachan Area, Alberta.
Canadian Forestry Service, Dept. Env. Northern Forest Res. Center,
Edmonton, Albt., Info. Rpt. NOR-X-74.
191
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APPENDIX B
HONEYBEE COLLECTION SITES IN THE FORT UNION BASIN
Site No. Location Site No. Location
BN-1 T3N, R41E, SEC 28 BSE-6 T2S, R46E, SEC 34
BN-2 T4N, R40E, SEC 26 BSE-7* T2S, R47E, SEC 19
BS-1 TIN, R41E, SEC 24 BSE-9 T4S, R51E, SEC 25
BS-4 T1S, R42E, SEC 8 BSE-9a* T4S, R51E, SEC 30
BS-5 T1S, R42E, SEC 3 BSE-10 T8S, R51E, SEC 20
BSE-8 T3S, R45E, SEC 18 BE-1 T2N, R43E, SEC 16
BSE-11 T7S, R47E, SEC 9 BNE-2 T3N, R43E, SEC 33
BSE-12 T7S, R46E, SEC 6 BNE-2a T3N, R43E, SEC 21
BSW-] T1S, R41E, SEC 13 BNE-3 T3N, R43E, SEC 8
BSW-2 T1S, R41E, SEC 22 BNE-4 T4N, R43E, SEC 32
BSW-3 T2S, R41E, SEC 4 BNE-5 T4N, R42E, SEC 13
BSE-1 TIN, R42E, SEC 26 BNE-6 T5N, R42E, SEC 5
BSE-2 TIN, R43E, SEC 19 BNE-10 T6N, R42E, SEC 16
*Wild honeybee colonies.
192
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APPENDIX C
DIRECTIONS FOR PONDEROSA PINE-INSECT STUDY
I. Field Procedures
A. Record the plot location, date of collection, and number of the
tree being sampled in the field diary.
B. Prepare a collection card with the same information as in the
previous step, and place the card in a new plastic collection bag.
C. Classify the tree based on condition (modification of the procedure
of Carlson et aj., 1974), and record in diary.
Tree Class
0 No apparent insect feeding or pollution damage. Needle
retention appears normal.
1 Some damage evident: light insect activity, foliar burn
restricted to older needles, or crowns becoming thin.
Needle retention below normal.
2 Damage by insect feeding quite evident, foliar burn con-
spicuous, or thin crown. Needle retention below normal
or poor.
3 Heavy insect damage evident. Excessive insect feeding
and/or foliar burn. Crowns thin and needle retention
poor.
4 Tree is dead or dying.
D. Estimate height of tree and measure diameter at breast height;
record in field diary.
E. Inspect the base, trunk, crown, branches, and twigs of the tree
for insects and injury (includes damage by storms, porcupines, or
other agents). Record observations and place any insects into a
labeled vial filled with 80% ethanol to which has been added a
drop of glycerol.
193
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APPENDIX C (continued)
F. Beat both the upper and lower foliage of the tree with a sweep net
(50 sweeps/level). Preserve any insects obtained in alcohol
(paper Lepidoptera); list any insects in field diary. If insect
has not been identified, give a brief written description and
indicate order or family.
G. Examine 100 mature cones (fewer if necessary) on the tree and
record the percent of the cones dwarfed or damaged by insect
feeding; identify the damaging insect(s).
H. Remove four branches (five internodes/branch) from top one half
and four branches from the lower one half of the foliage from side
of tree facing Colstrip. Place the branches from each of the two
levels in separate plastic bags. Keep the samples out of the sun,
covered by a tarp or in a canvas bag. Process or refrigerate as
soon as possible.
I. If any dead or dying trees are visible from the marked sample
tree, inspect the trees and record the probable cause(s) of death.
II. Laboratory Procedures for each Sample
A. Cut branches from a given tree into internode segments to separate
the foliage by age.
B. From the four most recent ages, select four internodes for deter-
minations of percent needle retention.
C. Remove all fascicles end maintain by age.
D. Randomly select 100 fascicles of each age, remove the sheaths,
and save 100 needles.
E. For each of these 100 needles, inspect for each of the various
types of injury; including basal necrosis, basal scale (unidenti-
fied), pine needle scale (Phenacaspis pinifoliae, defoliators, tip
necrosis, chlorotic mottle, and weevil) and record the percentage
of needles affected by each agent.
F. Save representative specimens of different pathology types; kill
and fix in FAA (formalin-aceto-alcohol).
G. Save any insects in 80% ethanol, label, record any comments.
H. Complete measurements of cross-sectional area, needle length, and
moisture percentage.
I. Prepare all stripped needles for chemical analyses.
194
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APPENDIX D
CHECKLIST OF PREVALENT INSECTS ASSOCIATED
WITH PONDEROSA PINES IN SOUTHEASTERN MONTANA
Order
Family
Species
Neuroptera
Homoptera
Hemiptera
Coleoptera
Mantispidae
Chrysopidae
Aphidae
Coccidae
>i n
Coreldae
Buprestidae
Cerambycidae
Coccinellidae
Curculionidae
ClimacieTIa brunnea (Say)
Chrysopa sp.
Cinara ponderosae (Wms.)
Matsucoccus secretus Morrison
Phenacaspis pinifoliae (Fitch)
Leptoglossus sp.
Bupresti s spp.
Arthopalus foreicol 1 is (Lee. )
Batyle Thomson sp.
Monochamus maculosus Hald.
Neoclytus muricatul us (Kby.)
Prionus imbricorm's (L. )
Anatis quindecimpunctata (01iv. )
Cleis pi eta (Rand. )
Hippodamia convergens Guerin
Neomysia Casey sp.
Hagdalis Germar spp.
Pissodes probably fasciatus Hopk,
195
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APPENDIX D (continued)
Order Family
Coleoptera Curcul ionidae
n n n ii
" " Scarabaeidae
n n n M
" " Scolytidae
n n n n
n n n n
n n n n
n n n n
n n n n
n n n n
n n n n
n n n n
Diptera Cecidomyi idae
" " Tachinidae
Lepidoptera Arctiidae
n n M n
" " Gel echi idae
" " Geometridae
" " Lasiocampidae
" " Olethreutidae
ii ii n n
n n n n
" " Pyralidae
Species
Scythropus elegans (Couper)
Scythropus probably albidus Fall
Cotalpa lanigera Wick.
Polyphylla decimlineata (Say)
Conophthorus Hopk. probably
ponderosae
Dendroctonus ponderosae Hopk.
" " valens LeConte
Dryocoetes probably confuses Sw.
Hylurgops subcostulatus (Mann.)
Ips calligraphus (Germar)
Ips p i ni (Say)
Pityophthorus sp.
Scolytus sp.
Unidentified species
Euphorocera sp. near edwardsii
(Williston)
Dasychira near grisefacta Dyar
Hypantria cunea (Drury)
Coleotechnites sp.
Phaeoura mexicanaria (Grote)
Malacosoma disstria Hbn.
Laspeyresia sp.
Petrova sp.
Rhyacionia (Busck) sp.
Dioryctria abietella (D.&S.)
196
-------
APPENDIX D (continued)
Order Fami 1y
Species
Lepidoptera
Hymenoptera
Pyralidae
Pieridae
Yponomeutidae
Diprionidae
ii ii
Ichneumonidae
Pteromalidae
Vespidae
Dioryctri'a spp.
Neophasia menapia Felder
Zellaria probably haimbachi
Busck
Dlprion sp.
Neodiprion fulvlceps complex
Ichneumon pulcherior (Heinrich)
Coeloplsthia suborbicularis
(Provancher)
Vespula spp.
197
-------
APPENDIX E
PRE-OPERATIONAL ON SITE MEASUREMENTS
Diameter % Dwarfed Tree Tree Porcupine
F Number Site
F-1300 N #4
F-1300.5
F-1305
F-1305.5
F-1310
F-1310. 5
F-1315
F-1315. 5
F-1320 "
F-1320.5
F-1850 NE #1
F-1850. 5
F-1855
F-1855.5
F-1860
Approx.
Height
(ft)
22
24
18
27
24
30
27
23
25
24
42
42
43
41
24
Breast
Height
(in)
11.9
10.8
14.6
12.6
18.0
17.7
15.4
15.3
12.1
11.1
21.3
15.9
15.8
18.1
11.1
Cones Class Class Attack
(50-100 (0-4) (0-4) (trunk and/
cones/tree) 1974 1975 or limbs)
2
7
3
0
32
0
6
2
3
0
20
18
20
34
42
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
NE*
Pt
NE
NE
NE
NE
NE
NE
RJ
P
NE
KE
NE
NE
NE
*Not evident
tPrevious
tRecent (within last 12 months)
198
-------
APPENDIX E (continued)
F Number
Approx.
Height
Site (ft)
Diameter % Dwarfed Tree Tree Porcupine
Breast Cones Class Class Attack
Height (50-100 (0-4) (0-4) (trunk and/
(in) cones/tree) 1974 1975 or limbs)
F-1860.5 NE #1
F-1865
F-1865.5
F-1870
F-1870.5
F-1575 NE #3
F-1575.5
F-1580
F-1580.5 "
F-1585
F-1585.5
F-1590
F-1590.5
F-1595
F-1595.5
F-1775 NW #3
F-1775.5
F-1780
F-1780.5
F-1785
F-1785.5
30
45
45
27
30
36
29
38
32
23
20
19
30
23
20
45
42
39
32
40
50
14.3
16.7
16.4
11.0
11.5
16.6
14.8
11.9
15.4
9.5
9.2
10.5
11.9
11.6
9.4
21.6
20.7
16.6
14.3
17.0
20.2
37
10
14
26
26
3
1
3
5
0
0
0
0
0
0
76
-
45
60
68
-
0
0 0
0
0 0
0
0 0
0
0 0
0
0 1
1
0 0
1
0 1
1
0 0
0
0 0
1
1 0
0
NE
NE
NE
P
NE
R
R
P
R
P
P
R
NE
NE
P
R
P
P
P
NE
NE
199
-------
APPENDIX E (continued)
F Number
Approx.
Height
Site (ft)
Diameter % Dwarfed Tree Tree Porcupine
Breast Cones Class Class Attack
Height (50-100 (0-4) (0-4) (trunk and/
(in) cones/tree) 1974 1975 or limbs)
F-1790 NW #3
F-1790.5 "
F-1795 "
F-1795.5
F-1350 NW #4
F-1350.5
F-1355
F-1355.5
F-1360
F-1360.5
F-1365
F-1365.5
F-1370
F-1370.5
F-1750 W #3
F-1750.5
F-1755
F-1755.5
F-1760
F-1760.5
F-1765
33
35
37
41
28
29
44
26
32
31
34
20
38
28
35
32
42
31
54
48
26
14.0
11.1
20.1
18.8
10.2
11.3
20. 1
13.2
15.3
15.9
14.3
17.3
20.1
16.2
13.7
12.6
19.4
13.5
10.2
10.5
10.0
60
62
73
37
-
30
-
13
55
48
44
60
33
25
-
23
32
-
0
-
0
-
1
-
1
-
1
-
1
-
0
-
0
-
0
0
-
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
0
0
1
0
p
p
p
p
NE
p
NE
P
NE
NE
NE
NE
NE
NE
NE
NE
R
P
P
NE
NE
200
-------
APPENDIX E (continued)
Approx.
Height
F Number Site (ft)
Diameter % Dwarfed Tree Tree Porcupine
Breast Cones Class Class Attack
Height (50-100 (0-4) (0-4) (trunk and/
(in) cones/tree) 1974 1975 or limbs)
F-1765.5 W #3
F-1770
F-1770.5 "
F-1200 W #4
F-1200.5
F-1205
F-1205.5
F-1210
F-1210.5
F-1215
F-1215.5
F-1220
F-1220.5
F-1475 SE #1
F-1475.5 "
F-1480 "
F1480.5 "
F-1485 "
F- 1485. 5
F-1490
F-1490.5
25
40
30
28
26
36
30
28
24
42
20
28
30
29
30
30
32
26
28
26
20
13.2
13.8
12.3
13.5
9.9
14.0
11.0
11.5
11.5
18.1
15.9
14.6
12. 1
12. 1
13.2
15.9
12.3
13.5
14.0
10.5
8.6
64
24
-
2
0
12
3
5
3
10
3
0
5
2
1
22
21
77
96
28
29
0
0 0
-
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
0 0
0
NE
p
NE
NE
NE
NE
NE
NE
NE
NE
NE
P
NE
NE
NE
NE
NE
NE
NE
NE
NE
201
-------
APPENDIX E (continued)
F Number Site
F-1495 SE #1
F-1495.5
F-1050 SE #3
F-1050.5
F-1055
F-1055.5
F-1060
F-1060.5
F-1065
F-1065.5
F-1070
F-1070.5
F-1100 S #3
F-1100.5
F-1105
F-1105.5 "
F-1110
F-1110.5
F-1115
F-1115.5
F-1120
Approx.
Height
(ft)
26
26
30
35
38
35
26
33
26
26
36
33
42
31
30
34
28
35
45
29
34
Diameter
Breast
Height
(in)
18.8
12. 1
13. 1
17.3
19.3
15.6
14.5
18.1
11.5
14.6
16.2
18.0
15.8
9.9
12.1
13.5
12.7
15.1
16.8
11.1
19.1
% Dwarfed
Cones
(50-100
cones/tree)
26
26
9
10
4
6
10
8
4
10
12
-
-
-
-
-
-
-
-
-
-
Tree
Class
(0-4)
1974
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
0
-
0
Tree
Class
(0-4)
1975
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Porcupine
Attack
(trunk and/
or limbs)
NE
NE
P
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
202
-------
APPENDIX E (continued)
Diameter % Dwarfed Tree Tree Porcupine
F Number
F-1120.5
F-1825
F-1825.5
F-1830
F-1830.5
F-1835
F-1835.5
F-1840
F-1840.5
F-1845
F-1845.5
F-1275
F-1275.5
F-1280
F-1280.5
F-1285
F-1285.5
F-1290
F-1290.5
F-1295
F-1295.5
Approx.
Height
Site (ft)
S #3 32
E #1 45
51
45
42
24
27
32
25
38
34
E #3 24
25
30
24
38
37
27
37
35
33
Breast
Height
(in)
9.9
15.8
14.0
25.1
20. 1
15.0
13.0
20.0
11.0
20.3
13.7
16.1
14.2
17.8
18.3
14.8
15.6
13.7
15.9
20. 1
16.6
Cones
(50-100
cones/tree)
-
29
31
32
46
41
39
47
62
30
94
5
0
1
6
3
1
2
2
0
1
Class
(0-4)
1974
-
0
-
0
-
0
0
-
0
-
0
-
0
-
0
-
0
0
-
Class
(0-4)
1975
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Attack
(trunk and/
or limbs)
NE
NE
NE
R
NE
NE
NE
R
R
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
NE
203
-------
APPENDIX E (continued)
F Number
Approx.
Height
Site (ft)
Diameter % Dwarfed Tree Tree Porcupine
Breast Cones Class Class Attack
Height (50-100 (0-4) (0-4) (trunk and/
(in) cones/tree) 1974 1975 or limbs)
F-1250 E #4
F-1250.5
F-1255
F-1255.5
F-1260
F-1260.5
F-1265
F-1265.5
F-1270
F-1270.5
F-1225 E #5
F-1225.5
F-1230
F-1230.5
F-1235
F- 1235. 5
F-1240
F-1240.5
F-1245
F-1245.5
26
18
28
30
30
24
15
17
18
16
36
33
33
25
37
40
29
34
35
28
21.0
9.5
14.3
11.6
12.3
12. 1
9.4
13.2
10.7
8.9
21.3
21.1
20.7
12.7
23.6
18.3
11.6
18.8
26.1
17.2
0
3
3
6
3
2
0
1
0
0
25
20
3
3
4
3
4
5
5
6
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
-
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
p
p
p
R
p
p
R
p
p
R
R
R
P
R
P
P
R
NE
R
R
204
-------
APPENDIX F
BIBLIOGRAPHY TO THE MATSUCOCCUS
(Homoptera, Coccoidea, Margarodidae)
Carle, P., J. P. Carde, and C. Boulay. 1970. Comportement de piqure de
Matsucoccus feytaudi Due. (Coc. Margarodidae) caracterisation histo-
logique et histochimique des desorganisations engendrees dans le
vegetal (Plnus pinaster Ait. var. mesogeensis). Ann. Sci. Forest
27(1):89-104~:
Herbert, F. B. 1921. The Genus Matsucoccus with a new species. (Hemip. -
Homop.). Proc. Entomol. Soc. Washington 23:15-22.
Keen, F. P. 1928. Insect enemies of California pines and their control.
Calif. Div. Forestry. Bui. 7, 113pp.
Keen, F. P. 1952. Insect Enemies of Western Forests. Revised ed. , U.S.D.A.
Misc. Publ. 273. 280 pp.
McCambridge, W. F. and D. A. Pierce. 1964. Observations of the Life
History of the Pinyon Needle Scale, Matsucoccus acalyptus (Homoptera,
Coccoidea, Margarodidae). Ann. Ent. Soc. Am. 57:197-200.
McKenzie, H. L. 1941. Injury by Sugar Pine Matsucoccus Scale Resembles
That of Blister Rust. Jour. For. 39:488-489.
1941. Matsucoccus bisetosus Morrison, a Potential Enemy
of California Pines. Jour. Econ. Ent. 34:783-785, illus.
. 1943. The Seasonal History of Matsucoccus vexillorum
Morrison (Homoptera: Coccoidea: Margarodidae). Microentomology 8
(Part 2):42-52, illus.
, L. S. Gill and Don E. Ellis. 1948. The Prescott Scale
(Matsucoccus vexi1lorum) and Associated Organisms that Cause Flagging
Injury to Ponderosa Pine in the Southwest. Jour. Agr. Res. 76:33-51,
illus.
Schvester, D. 1974. Bio-ecologie des Matsucoccus (Coccidae Margarodidae)
en particulier de Matsucoccus feytaudi Due. In: Ecologie Forestiere.
P. Pesson, comp. 241-256.
205
-------
Siewniak, M. December, 1971. Uszkadzanie sosny pospolitej (Pinus silves-
tris) przez czerwca korowinowca (Matsucoccus p i m' Green 1925; Margarodi-
dae, Coccoidea). Sylwan. 115(12):35-41.
. 1972. Potrzeby uwzglednienia zjawiska sprzezenia zwrotnego
w badaniach entomologicznych ukladu pasozyt-zywiciel. Ekol. Pol., Ser.
B. 18(l):29-37.
Taketani, A. 1972. Studies on a margarodid scale, Matsucoccus matsumurae
(Kuwana) (Hemiptera; Coccoidea). Jap. Forest Exp. Sta. Bui. 246:1-9.
Washburn, R. L. 1962. Forest Insect Condition in the United States, 1961.
Intermountain States. U.S. Forest Service, Wash., D.C. 16-20.
206
-------
APPENDIX G
TABLE G.I. PEARSON PRODUCT-MOMENT CORRELATION COEFFICIENT FOR PPM FLUORIDE
IN ADULT WORKER HONEYBEES VERSUS POLLEN, 1975 AND 1976 DATA
N = 114, Chi Square = 3.841, F-Value = 3.0700000, Alpha = .0500
The Transformation Codes are 0 and 0
Yl Y2
(Honeybees) (Pollen)
Mean 9.57 1.89
Variance 16.311 0.840
Standard Deviation 4.0386 0.9167
Standard Error 0.3782 0.0858
Covariance -0.395
The product-moment correlation coefficient is -0.10674.
The 95.0 percent confidence limits are LI = 0.2851 and L2 = 0.0788.
The eigenvalues are 16.32101 and 0.83035.
The equation of the principal axis is: Yl = 83.89246 + -39.17078 Y2.
The equation of the minor axis is: Yl = 9.52261 + 0.02553 Y2.
The 95.0 percent confidence limits to the slope of the principal axis are:
LI = 55.20636 L2 = 14.43632
Following Pairs Show the Coordinates of
A to H for Plotting Confidence Elli
Number One Yl
A
B
C
D .
E
F
G
H
10.5071
8.6350
9.5711
9.5711
8.6296
10.5125
9.5765
9.5656
Points
pse.
Y2
1.8974
1.8974
2.1099
1.6849
1.9214
1.8733
2.1097
1.6850
207
-------
TABLE G.2. PEARSON PRODUCT-MOMENT CORRELATION COEFFICIENT FOR PPM FLUORIDE
IN ADULT WORKER HONEYBEES VERSUS POLLEN, 1975 DATA
N = 66, Chi Square = 3.841, F-Value = 3,1400000, Alpha = .0500
The Transformation Codes are 0 and 0
Yl Y2
(Honeybees) (Pollen)
Mean 8.13 1.90
Variance 7.346 0.776
Standard Deviation 2.7103 0.8806
Standard Error 0.3336 0.1084
Covariance 0.305
The product-moment correlation coefficient is: 0.12801.
The 95.0 percent confidence limits are LI = 0.1177 and L2 = 0.3590.
The eigenvalues are: 7.36038 and 0.76134.
The equation of the principal axis is: Yl = 32.91300 + 21.55098 Y2.
The equation of the minor axis is Yl = 8.22019 + -0.04640 Y2.
The 95.0 percent confidence limits to the slope of the principal axis are:
LI = 7.47355 L2 = 24.81377
Following Pairs Show
A to H For Plotti
Number Two
A
B
C
D
E
F
G
H
the Coordinates of Poi
ng Confidence Ellipse
Yl
8.9675
7.2962
8.1318
8.1318
8.9743
7.2893
8.1192
8.1444
nts
Y2
1.9045
1 . 9045
2.1761
1.6330
1.9436
1.8655
2.1755
1.6336
208
-------
TABLE G.3. PEARSON PRODUCT-MOMENT CORRELATION COEFFICIENT FOR PPM FLUORIDE
IN ADULT WORKER HONEYBEES VERSUS POLLEN, 1976 DATA
N = 48, Chi Square = 3.841, F-Value = 3.1900000, Alpha = .0500
The Transformation Codes are 0 and 0
Yl Y2
(Honeybees) (Pollen)
Mean 11.55 1.88
Variance 22.148 0.948
Standard Deviation 4.7061 0.9736
Standard Error 0.6792 0.1405
Covariance -1.338
The product-moment correlation coefficient is: -0.29208.
The 95.0 percent confidence limits are LI = -0.5321 and L2 = 0.0086.
The eigenvalues are: 22.23181 and 0.86378.
The equation of the principal axis is: Yl = 41.56816 + 15.90366 Y2.
The equation of the minor axis is: Yl = 11.43132 + 0.06288 Y2.
The 95 percent confidence limits to the slope of the principal axis are:
LI = 210.50580 L2 = 8.23499
Following Pairs
A to H For
Number One
A
B
C
D
E
F
G
H
Show the Coordinates of Points
Plotting Confidence Ellipse
Yl Y2
13.2087
9.8913
11.5500
11.5500
9.8158
13.2842
11.5715
11.5285
1.8875
1.8875
2.2306
1 . 5444
1.9965
1.7785
2.2293
1.5457
209
-------
APPENDIX H
MEAN FLUORIDE IN ADULT WORKER HONEYBEES
1975-1976.
August 1975
Each value represents
a different hive
Site
SW1
SW2
SW3
SI
S4
S5
SET
SE2
NE2
NE3
NE4
N2
NE10
SE11
S9
SE10
SE12
SE9
NE5
NE2a
SE6
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
Bees
Pollen
1
8.6
1 .5
7.6
1 .1
8.5
6.3
2.9
7.2
0.5
8.8
2.3
8.2
1.7
7.8
1 .1
6.5
1.2
6.8
1 .3
6.6
1 .9
4.5
1.5
55.7
2.9
8.5
1.0
10.6
2.4
5.6
1 .8
9.8
1.6
10.2
1 .3
6.2
2.0
11 .1
10.6
2
4.7
1 .5
6.2
1.8
10.7
8.9
4.1
6.2
0.6
7.0
2.5
9.7
1.5
6.3
1 .1
7.5
2.5
8.9
3.2
1 .4
4.6
2.0
58.8
1 .5
11 .1
1.2
15.2
3.0
6.7
2.6
10.6
1.8
7.2
1.2
6.3
2.6
6.9
18.3
3
9.9
2.2
5.3
2.3
7.5
6.3
2.3
8.1
0.4
7.6
2.4
10.0
1.4
5.4
1.9
8.1
1.5
9.6
1.9
9.1
1.9
3.4
2.1
33.5
1.9
7.8
1.6
20.5
2.8
10.5
3.6
7.2
1.7
12.1
1.3
8.4
1 .8
7.4
8.2
4
9.3
2.0
12.2
2.8
7.9
5.1
2.9
9.7
0.9
5.4
2.2
8.7
1.4
10.4
1 .6
9.7
1.4
8.5
2.4
6.9
0.6
6.2
1.4
44.9
1 .6
--
10.5
3.1
6.2
2.0
8.0
1.6
11.2
1.8
7.8
0.6
7.3
11.4
Each value
represents
a combined
sample from
8-12 hives
5
8.5
8.5
10.0
8.2
9.7
9.3
12.0
9.7
10.2
9.2
8.1
7.1
47.6
--
16.5
8.3
7.8
10.6
9.3
10.6
6
8.3
8.2
10.9
8.8
9.6
8.4
9.7
11.2
8.5
8.6
7.0
6.3
60.7
--
19.2
7.4
9.4
9.9
6.9
8.2
August
September 1976
Each value represents
a different hive
1
8.8
2.2
12.0
5.0
6.7
2.1
6.4
2.5
7.8
1.7
15.2
2.7
15.8
1.2
6.7
0.8
16.7
1.6
12.6
1.6
16.2
2.1
3.9
2.3
77.6
2.9
3.8
0.6
12.5
2
11.8
2.5
10.2
1.1
8.9
1.7
5.8
2.9
8.0
2.2
10.4
2.2
17.1
0.6
8.4
1.2
15.4
1.4
14.4
0.8
16.8
1 .2
4.0
110.8
2.7
5.9
2.8
20.6
3
7.8
1.2
7.8
1 .5
14.2
0.7
5.8
3.3
11.4
2.8
16.2
2.1
14.7
0.7
13.1
1.0
21.5
1.3
23.1
1.5
10.4
0.9
3.0
99.8
1.9
4.0
3.4
--
4
13.1
2.6
9.9
1.7
9.1
1.3
5.9
4.1
12.4
1.5
15.4
3.0
20.2
0.8
10.5
0.8
17.5
1.5
13.8
2.8
11.0
3.1
--
104.8
1.7
3.2
--
Each value
represents
a combined
sample from
8-12 hives
5 6 H20
0.6
10.1 8.4 0.2
11.0 10.6 0.6
5.7 6.3 0.1
13.6 12.1 0.6
12.0 13.3 0.6
11.0 10.5 0.6
12.4 13.5 0.6
15.8 14.1 0.3
11.7 11.7 0.6
12.5 12.7 0.6
1.7 2.3 0.1
98.3 95.4 10.5
—
210
-------
APPENDIX I
TABLE I.I. BASIC STATISTICS FOR FLUORIDE IN ADULT WORKER HONEYBEES, 1975.
N
Mean
Median
Variance
Stand. Dev.
Coeff. Var.
= 79
Statistic
8.36
8.00
7.88
2.80
33.55
0 Classes
Standard Error
0.315
0.395
2.954
Transformation Code = 0
Confidence Limits
95%
7.73 8.99
7.21 8.78
27.67 39.43
TABLE 1.2.
BASIC STATISTICS
FOR FLUORIDE IN
ADULT WORKER HONEYBEES, 1976.
N
Mean
Median
Variance
Stand. Dev.
Coeff. Var.
= 53
Statistic
11.27
11.00
25.39
5.03
44.67
0 Classes
Standard Error
0.692
0.867
5.133
Transformation Code = 0
Confidence Limits
95%
9.88 12.67
9.24 12.74
34.36 54.99
F = 3.2216, which is highly significant.
The exact probability of F = 3.2216 with 78 and 52 D.F. is 0.00003; therefore
the variances for 1975, 1976 are unequal. (H.:af ^ a|).
211
-------
APPENDIX I (continued)
TABLE 1.3. PPM FLUORIDE IN ADULT WORKER HONEYBEES, 1975 AND 1976, TWO LEVEL
NESTED ANOVA.
Level
2 (years)
1 (sites)
0 (hives/site)
Level
2
1
0
*Exact probability
** Exact probability
SS
269.004
1339.043
596.458
Anova Table
DF MS FS
1 269.003 6.3928*
32 1.845 6.8753**
31.9 42.079
98 6.086
VARIANCE COMPONENTS
Variance Component Percent
3.
9.
6.
of FS = 6.3928; DF
of FS = 6.8753; DF
577 18.947
215 48.813
086 32.238
1, 31.9; is 0.0125.
32, 98; is less than 0.001.
TABLE 1.4. BASIC STATISTICS FOR FLUORIDE IN ADULT WORKER HONEYBEES.
1975 VS. 1976
(Approximate test of equality of means
when variances are heterogenous.)
Sample Number
(1975) 1
(1976) 2
Mean
8.36
11.27
Variance
7.883
25.396
F* = 14.648
DF = 1 and 73.8
t = 3.83
F* is greater than F ^^ ?3 ^ = 11.9; we reject the null hypothesis that
samples were drawn from populations with equal mean fluoride content.
The exact probability of F = 14.648 with (1, 73.8) DF is 0.00051.
212
-------
SECTION 6
THE EFFECTS OF* COAL-FIRED POWER PLANT EMISSIONS ON VERTEBRATE
ANIMALS IN SOUTHEASTERN MONTANA (A REPORT OF PROGRESS)
by
R. A. Lewis, M. L. Morton, M. D. Kern, J. D. Chilgren, and E. M. Preston
INTRODUCTION
Vertebrate responses to air pollution vary seasonally with the quality and
quantity of ecosystem resources, with the sex, age, or physiological state of
the organism, and with secondary stressors, such as disease, competitive
interactions, or other pollutants. Hence, field work in this project has em-
phasized the description and evaluation of annual and life cycles and the
elucidation of mechanisms that regulate these cycles.
The preoperational (baseline) phase of the investigation is nearing com-
pletion, although analyses of this work will continue for some time. Major
objectives for this phase of the study are to:
1. Measure and predict changes in population structure and/or dynamics
of grassland birds and small mammals as a function of annual, seasonal, and
life cycles as well as other environmental information including biotic
interactions and physical factors that influence the structural and
dynamic processes under study;
2. assess the specific effects of air pollution, if possible, upon avian
and mammalian population structure and dynamics as well as upon specific
organ systems, and
3. evaluate selected physiological, biochemical, and behavioral
functions that may have potential for sensitive assay of pollution
challenge. Hopefully, low levels of pollution stress will be identified
before serious or irreversible effects occur.
Although considerably less substantial than that for birds, the baseline
work on mammals is more complete at this time and is therefore emphasized in
this synopsis. The preoperational components of the study will be completed for
mammals in 1977, but not until 1978 for birds. The bulk of the remaining bird
work will be performed under grant by appropriate investigators. The major
components are:
1. Reproductive and developmental biology;
213
-------
2. measures of condition, physiologic stress, homeostasis, and adapta-
tion;
3. population biology, and
4. histological cycles of organ-systems of potential or probable con-
cern.
Field experiments, now completed, on the effects of sulfur dioxide on the
population movements and physiology of small mammals will be treated in a
subsequent report.
Avian species being studied are the Western Meadow!ark (Sturnella neglec-
ta). Mourning Dove (Zenaidura macroura), Lark Bunting (Calamospiza melano-
corys), Vesper Sparrow (Pooecetes gramineus), and Lark Sparrow (Chondestes
grammacus). The mammals of greatest importance are the Deer Mouse (Peromyscus
maniculatus) and Prairie Vole (Microtus ochrogaster).
The reproductive and developmental portions of the study emphasize growth
and development of the young, bioenergetics, productivity, and the regulation of
reproductive processes and molt. The physiological component treats those
functions that reflect condition and vigor of the species and their stress
responses. Population parameters, such as abundance and dispersion of species,
sex, and age ratios, and some of the mechanisms that effect population adjust-
ments are also of concern. Finally, the study of histological cycles is
designed to help interpret mechanisms at all levels and to identify direct and
indirect (e.g., intercurrent disease) effects of pollution.
MAMMALS
INTRODUCTION
The study of small mammals is largely confined to the mice and voles
(Cricetidae). These are the most abundantly distributed and easily trapped
mammals in southeastern Montana. Some information is also being gathered on
members of the families Heteromyidae (including pocket mice), and Sciuridae
(squirrels, chipmunks, etc.).
The use of small mammals as indicator species of air pollution is not new
(e.g., Kay, 1974), but it has not received much attention. Although the impact
of small mammals on the character of grassland ecosystems is reasonably well
documented (Golley et al_. , 1975), their potential importance with respect to
ecosystem responses to air pollution has rarely been considered. Small mammals
in the Northern Great Plains contribute to maintenance of the health and vigor
of the grasslands. Mechanisms include the following:
1. Grazing, within limits, promotes the growth of plants. Thus, grazing
by rodents may have a significant effect on the productivity of the
range.
2. Burrows of fossorial mammals may increase water storage and retention
in the soi1.
214
-------
3. Burrowing increases transport of minerals from deeper to upper soil
layers and may increase the rate of decomposition of organic matter.
4. Feces may enrich soil biota.
5. Specific effects depend in large part upon habits and population
structure of the predominant species in a community. For example,
some species consume insects (many of which are harmful from man's
point of view) or weed seeds, whereas others can damage agricultural
crops.
6. Rodents are important food of many avian and mammalian predators.
Many changes in rodent population structure would be expected to have an
appreciable impact upon grasslands.
STUDY AREAS AND METHODS
The information on small mammals presented in this report was gathered at
selected sites east or southeast of Colstrip, Montana, between the spring of
1974 and the fall of 1976. Investigators were in the field each month. Thus
seasonal changes as well as instantaneous evaluations of many functions were
assessed. In 1976 only the McRae site remained under intensive investigation.
Five locations were selected, surveyed, and staked in mixed sagebrush-
grassland habitat in the spring of 1974. Four of these sites (Kluver North,
Kluver East, Kluver West, and McRae1s) were chosen because of their proximity
to exclosures constructed for grassland studies (see other reports, this vol-
ume). The remaining site was located on Pony Creek about one km west of
Gar-field's ranch house. Square grids 150 m on a side (2.25 ha) were con-
structed. Trapping stations were 15 m apart (121 stations per grid). Each
month during the 1974 and 1975 field seasons, Sherman live traps (7.6 x 7.6 x
25.4 cm) were baited with rolled oats for three or four nights with free entry
and exit (prebaiting). This was followed by a one-day capture, mark, and
release (CMR) trapping session. In 1976 the traps were prebaited for two
nights followed by three successive sessions of capture, mark, and release.
This procedure was repeated at three-week intervals.. Traps were set at dusk
and picked up at dawn. In cold weather tissue paper for nest material and large
quantities of bait were provided to enhance survival. The toes (no more than
one per foot) of trapped mice were clipped to permit individual identification.
Mice were sexed, aged, and weighed to the nearest 0.1 g on a pan balance. They
were also examined externally for signs of sexual activity and pelage changes.
All were released at their station of capture.
Specimens were collected at weekly intervals in live traps and transported
to the laboratory at Fort Howes Ranger Station. Linear dimensions (body, ear
to notch, hind foot, and tail) were measured with precision calipers.
Following ether anesthesia, two blood samples were taken from the orbital sinus
in heparinized capillary tubes. Hematocrit was determined from these samples
following high-speed centrifugation for 10 minutes. Hemoglobin concentration
was measured spectrophotornetrically by the cyanmethemoglobin technique, and
plasma protein concentration was measured with a diffraction meter. Heart,
215
-------
lungs, kidneys, adrenals, spleen, liver, and reproductive tracts were removed
and fixed in Bouin's solution or 10% neutral formalin, and after one week
weighed to the nearest 0.2 mg on a torsion balance. After weighing, tissues
fixed in Bouin's were transferred to 70% ethanol. Reproductive tracts
collected in September and October 1975 were supplied by Don Dodge from C.
Gordon's laboratory. Gonadal weights given are always those of the paired
glands.
It was noted that reproductive readiness, large testes, or pregnancy were
not observed in P. maniculatus weighing less than 14 g. Therefore, the term
"immature" is applied to animals below 14 g body weight and "adult" to those 14
g or heavier.
REPRODUCTIVE AND DEVELOPMENTAL BIOLOGY
The reproductive and developmental portions of the study emphasize de-
scription of the annual reproductive cycles of a small set of indigenous spe-
cies. The evaluations are not complete at this time.
Females
Pregnant deer mice occurred in the samples from early spring 1974 through
late summer 1975 (Table 6.1). The first two females trapped in March 1975 were
pregnant. Embryo lengths j_n utero were 60 mm and 110 mm, the latter probably
near term. In 1975 the proportion of pregnant females decreased in July and
August and in September only one of 20 had embryos. None of the 11 adults
trapped in October were pregnant. In September 1974, however, one of three
adults was pregnant and in late October four of five adults were pregnant,
although none of five taken in early October were pregnant. A resurgence of
breeding in October 1974 may have been related to unusually wet weather during
which there was new growth of grasses. Precipitation in October 1974 was more
than double that of the long term average.
Litter sizes of all species, based on dissections of pregnant females, are
presented in Table 6.2. In P_. maniculatus litter size (1975) based on counts
of uterine scars was not significantly higher (0.10 < P < 0.20) than counts of
visible embryos. Similarly, litter size did not vary significantly throughout
the season.
Litter sizes between 1975 and 1976 differed significantly for *both the
deer mouse (P < 0.05) and the prairie vole. Data for the voles, however, is
based upon a small sample size and might merely reflect variation in area-
specific populations. Nearly all pregnant voles in 1976 were obtained from one
region of about 250 m2. One major difficulty in the study of the prairie vole
is its erratic distribution.
A chi-square test revealed no difference in distribution of embryos be-
tween the two uterine horns (P > 0.98) in P. maniculatus.
The weight of the reproductive tract of female mice changes seasonally in
association with the production of young. Thus, in P_. maniculatus there wso
about a four-fold difference between monthly mean ovarian weight in winter and
216
-------
TABLE 6.1. PERCENT OF PREGNANT FEMALES IN SAMPLES OF Peromyscus maniculatus
COLLECTED NEAR COLSTRIP.
Month
July 1974
August
September
October
November
December
January 1975
March
April
May
June
July
August
September
October
April 1976
May
June
July
August
September
Number in
Sample
4
8
14
10
8
10
3
2
9
6
8
9
10
20
11
18
4
13
7
10
3
Percent
Pregnant
75.0
62.5
28.6
40.0
0.0
0.0
0.0
100.0
77.8
66.7
75.0
33.3
40.0
5.0
0.0
33.3
25.0
46.2
57.1
40.0
33.3
TABLE 6.2. LITTER SIZE
IN MICE COLLECTED NEAR COLSTRIP.
Species
Peromyscus maniculatus
Microtus ochrogaster
Perognathus fasciatus
Reithrodontomys megalotis
Size
1975
1976
1975
1976
1975
1975
1976
1975
1976
1975
determi nant
uterine scars
uterine scars
embryos
embryos
total
embryos
embryos
embryos
embryos
+ 1976
N
9
2
42
22
24
11
6
1
2
2
Mean
5.
6.
4.
5.
5.
3.
5.
6.
5.
4.
33
50
95
68
75
00
00
00
00
00
0
0
0
0
0
0
SD
.87
--
.99
.72
.74
.85
.89
--
--
--
Range
4-
6.
3.
6
7
7
4-7
4-
2-
4-
--
--
--
7
4
6
-
-
-
217
-------
summer and a 50-fold difference in uterine weight (Figure 6.1). The largest
average ovarian weights were recorded in April and May and the lowest in
November, December, and January. Mean uterine weight in October 1975 was
strongly biased by one female with seven near-term implanted embryos, whose
uterus weighed 11.9 g. Mean uterine weight of the other 19 females in the
sample was only 29.8 mg.
Ovarian weight in £. maniculatus varied seasonally and in proportion to
body weight (P < 0.005). The least squares regression line of ovarian weight
on body weight in pregnant females was Y = 3.13 ± 0.69X (standard error of
estimate of Y on X = 3.72; r = 0.574).
M. ochrogaster is reproductively active throughout the year except in mid-
winter (see Table 6.3). A pregnant female and a male with scrotal testes taken
on December 5 had just reached sexual maturity, as judged by body weight, and
were probably engaged in their first breeding effort. The data do not permit
accurate determination of the annual reproductive period in M. ochrogaster, but
reproductive tracts were definitely reduced in size during winter months and
only one male with scrotal testes was collected from November through January
(Table 6.3).
Males
There is a marked seasonal variation in testicular weight of P. manicu-
latus (Figure 6.2). Maximum weight and presumably peak functional status were
maintained in the samples from March through August.
A cycle in seminal vesicle weight paralleled that of the testes during
phases of peak and decreasing testicular size but lagged during testicular
recrudescence. This expected relationship illustrates the trophic effect of
testosterone on sexual accessory organs (McKeever, 1964a, b; Cinq-Mars and
Brown, 1969; Chapman, 1972). The decision to assign adult status to £.
mam'culatus with a body weight of 14 g or more was based upon visual inspection
of reproductive tracts. Flake (1974) found minimum weight of pregnant £.
maniculatus to be 18 g, but investigators in this research found it to be at
least as low as 14.3 g. Individual males of intermediate size exhibited a
seasonal change in tendency to reach sexual maturity. Males in May and June
tended to have larger testes and seminal vesicles than individuals of com-
parable size later in the reproductive season in July and August (Figure 6.3).
Testicular weights differed significantly (P < 0.05) among all body weight
groups except 14.0 to 14.9 g, as did seminal vesicle weights (P < 0.01) in
animals weighing 16.0 g or more.
The breeding season in eastern Montana has not been previously documented
for any species of mouse, but in Colorado pregnant deer mice (£. maniculatus)
were recorded primarily from April through September (Lechleitner, 1969), and
in Wyoming from April through August (Brown, 1966). Data show that the repro-
ductive period of deer mice in the Col strip area extends from mid-March to mid-
September.
The distinct breeding cycle in £. mam'culatus results in pronounced and
regular variation in the number and proportions of immatures captured (Figures
218
-------
20
£
0
(2)
(8)
kf > 'J' (20,
Xi^t\l JI3)
\^ I
t
1.4
- 1,2
1.0
0,8
0,6
0,4
0.2
0.0
CD
k-
0>
5
Figure 6.1
JASONDJFMAMJJASO
Seasonal changes in mean ovarian weight (solid circles) and uterine weight (squares) in
Peromyscus maniculatus near Colstrip, Montana, July 1974 to October 1975. Vertical lines
indicated ± 2 SE for ovarian weight. Sample size for both organs shown in parentheses.
-------
500
ro
ro
CD
100
o>
E
-------
400
350
May,June
July, August
(23)
300
o>
E
(7)
100-5
a>
E
(/)
c
o
0>
Figure 6.3. Early (May, June) and late (July, August) summer differences in
testes weight (histograms) and seminal vesicles (triangles) in
young Peromyscus maniculatus. Sample size shown in
parentheses.
221
-------
TABLE 6.3. BODY AND REPRODUCTIVE ORGAN WEIGHT IN Hicrotus ochrogaster COLLECTED NEAR COLSTRIP, MONTANA
ro
no
July 1974 to August 1975.
July 28
August 9
August 20
September 3
November 27
December 5
December 17
Sex
1974 M
M
M
F
F
F
M
M
F
F
F
F
F
F
M
M
F
M
M
M
M
F
M
M
Body
Weight
g
25.4
23.4
28.8
28.3
13.3
34.2
38.9
30.5
28.4
44.5
34.1
42.3
19.1
24.5
26.0
23.7
28.9
21.0
31.7
26.8
25.3
24.4
36.2
20.3
*
Testes
mg
154.2
73.8
273. 81
--
--
--
534. 61
391. O1
--
--
--
--
--
--
82.4
66.8
--
18.8
124.8
38.0
230. 21
--
130.0
32.8
Ovaries
and uterus
mg
--
--
--
133.
34.
221.
--
--
384.
4450.
453.
1820.
14.
44.
_
--
130.
--
--
--
--
58.
--
--
8
8
0
0
O2
6
O2
8
4
6
0
January 9 1975
March 13
April 15
April 18
May 6
May 16
June 10
June 17
August 5
August 7
August 12
Sex
F
F
F
M
M
M
F
F
F
F
M
M
F
M
F
M
F
M
M
F
F
M
F
M
M
F
M
M
M
M
Body
Weight
g
25.6
29.5
30.9
22.8
34.0
22.9
25.4
23.6
24.2
27.8
36.0
28.3
38.0
36.2
28.0
35.1
33.3
30.3
30.8
32.7
27.1
31.2
39.2
32.0
37.2
40.3
37.6
31.0
20.7
23.6
Testes
mg
--
—
--
80.0
84.0
34.0
--
--
--
--
598. I1
288.4
--
710. 21
--
590. 21
--
387. 21
505.4
--
--
780. 31
--
104. 91
76. 91
--
920. 21
214. 61
362. O1
38.6
Ovaries
and uterus
mg
48.
58.
84.
--
—
--
94.
306.
72.
44.
--
--
1080.
--
200.
--
388.
--
--
408.
187.
--
207.
--
--
266.
--
--
--
0
0
8
6
72
0
8
O2
2
22
22
62
22
O2
xTestes in scrotum
2Embryos visible in uterus
-------
6.4 and 6.5). All deer mice captured in March were adults, but one caught on
April 18 was 12 g. In 1976 the smallest immatures captured in April were 7.0,
8.5, and 11.9 g captured on April 9, 13, and 23, respectively. The proportion of
immatures in the trapped sample increased thereafter, peaking in October of 1974
and September or thereafter in 1976. Immatures captured in mid-winter were
undoubtedly from the last litters of autumn.
During much of the reproductive period females are heavier than males
(Figure 6.6). This weight increase was undoubtedly associated with pregnancy.
No differences in linear body dimensions between adult males and adult non-
pregnant females were found during this time (Tables 6.4 and 6.5). When all
animals sampled for the duration of the study were compared, however, tail
length was significantly greater in females (t = 1.90, P < 0.05) whereas hind
foot length was considerably greater in males (t = 3.09, P < 0.005). The sexes
did not differ in body length (t = 0.98, P < 0.10) or in ear length (t = 0.87, P
< 0.10).
MEASURES OF CONDITION, PHYSIOLOGICAL STRESS, HOMEOSTASIS, AND ADAPTATION
The blood vascular system of mammals is complex but well characterized.
Blood carries erythrocytes that are involved in oxygen transport, leucocytes
that mediate responses to insult, hormones, enzymes and other proteins, and a
host of other chemicals that are characteristic of a given functional state.
Some of the most easily assessed characteristics include hematocrit (percentage
of packed red blood cells occupied by a given volume of whole blood), hemoglobin
content, erythrocyte count, and plasma protein analysis. The first three are
highly regulated in vertebrate vascular systems. Plasma protein varies more in
relation to other factors such as blood osmotic pressure and nutritional state.
Hematocrit, blood hemoglobin content, and plasma protein concentration were
measured in rodents trapped in all three years of the study. In addition, serum
acetylcholinesterase assays have been performed on blood samples taken in 1976.
The hematocrit ratio and hemoglobin concentration of M. ochrogaster were
higher in winter (December and January) than in summer (April to September),
whereas plasma protein concentration was higher in summer (Table 6.6). Blood
data for P. maniculatus were analyzed by sex in both winter (November to Jan-
uary) and summer (April to September) (Table 6.7). As in M. ochrogaster,
hematocrit tended to be higher in winter than in summer. Unlike M. ochrogaster,
however, hemoglobin of males was higher in summer than in winter (Table 6.7).
The data indicate that mean corpuscular hemoglobin concentration (MCHC)
decreased during winter (Table 6.8). There was a statistically significant
decrease in plasma protein concentration (PPC) during winter in both male and
female £. maniculatus (Table 6.7).
In 1976 seasonal changes in blood functions were not determined since the
sampling period spanned only six months (April to September). However, suffi-
cient samples were taken to determine further sexual differences in hematocrit,
hemoglobin, and plasma protein for both the deer mouse and prairie vole (Tables
6.9, 6.10, 6.11). In the vole, males had significantly higher values of hema-
tocrit and hemoglobin than females (Table 6.9). There was also a tendency for
higher plasma protein readings in males.
223
-------
50
40
o
g 30
«*-
O
p
. 20
1 io
E
0
rigure 6.4.
JASONDJFMAMJJA
Seasonal change in percent of Peromyscus maniculatus captured as immatures (body weight less
than 14 g) near Colstrip, July 1974 to August 1975.
-------
IV)
rvi
en
80
60
40
.c
o
o 20
c
OJ
O
I 20
40
60
iill Adults
Immatures
Males
(24)
(57) HZ! (82) (4|)
(78) "~
(36)
Figure 6. 5.
(29)
(49)
ill
Females
(32)
J
S 0
1974
N
M
A M
1975
Seasonal change in sex and age of Peromyscus maniculatus captured near Colstrip, July 1974
to August 1975. Sample size shown in parentheses.
-------
26
24
22
o»20
ro
01
o
GO
16
14
12
10
8
(4)
(37)
(29)
(18)
ji*—t—k\*\J II IVI %J I ^t ^
(16)
JASONDJF
M
A M
J A
Figure 6.6. Seasonal changes in mean body weight in live Peromyscus maniculatus. Vertical bars indicate
± SE. Sample size shown in parentheses.
-------
TABLE 6.4. SEASONAL CHANGES IN LINEAR MEASUREMENTS IN MILLIMETERS OF ADULT, NON-PREGNANT FEMALE
Peromyscus maniculatus CAPTURED NEAR COLSTRIP, MONTANA, July 1974 to August 1975.
IX)
ro
Body Length
July 1974
August
September
October
November
December
January 1975
April
June
July
August
Total Sample
Mean
89.8
89.9
84.8
81.0
85.1
79.7
70.0
88.7
83.0
88.7
86.5
86.4
S.D.
3.2
6.4
4.9
8.1
5.8
3.1
0.0
5.1
11.5
3.6
7.1
(N)
12
37
33
6
11
7
3
6
2
9
5
131
Tan"
Mean
63.7
63.9
61.5
56.3
57.0
56.0
48.0
58.2
65.0
58.7
58.0
60.9
1 Length
S.D.
4.0
4.1
5.4
6.2
4.9
4.2
2.0
4.3
5.7
8.6
3.0
5.9
(N)
12
37
33
6
11
7
3
5
2
9
5
130
Hindfoot Length
Mean
19.0
18.4
18.6
18.5
18.5
18.7
17.7
18.7
19.5
19.6
19.4
18.7
S.D.
0.9
1.2
1.2
1.0
0.7
0.5
0.6
0.5
—
0.5
0.5
1.0
(N)
12
37
33
6
11
7
3
6
2
9
5
131
Ear
Mean
16.0
15.5
15.5
15.8
16.7
15.3
14.3
16.3
17.0
16.8
16.6
15.8
Length
S.D.
1.2
1.2
1.4
1.3
1.2
0.5
0.3
1.2
1.6
0.7
1.2
(N)
12
38
33
6
11
7
3
6
2
9
5
132
-------
TABLE 6.5. SEASONAL CHANGES IN LINEAR MEASUREMENT IN MILLIMETERS OF ADULT MALE Peromyscus CAPTURED NEAR
ro
co
COLSTRIP, MONTANA
, July 1
Body Length
July 1974
August
September
October
November
December
January 1975
March
April
May
June
July
August
Total Sample
Mean
87.6
90.6
82.0
85.2
83.2
79.4
73.4
84.7
88.1
83.0
85.8
87.1
89.9
85.7
S.D.
4.2
5.0
5.5
5.9
4.4
4.9
5.8
6.3
7.0
3.4
5.3
6.0
«3.7
6.6
(N)
22
65
61
17
15
11
8
7
10
10
16
26
12
280
974 to August
Tail
Mean
62.0
63.1
60.4
58.2
53.9
56.3
52.1
57.3
58.7
58.4
61.3
57.9
58.1
59.8
1975.
1 Length
S.D.
4.2
4.2
4.7
5.9
5.9
7.4
5.5
4.8
5.1
4.7
4.9
3.8
4.2
5.5
(N)
22
65
61
16
15
11
8
7
10
10
16
26
12
279
Hindfoot Length
Mean
19.4
18.6
18.8
18.8
18.8
19.0
18.5
20.9
18.7
19.5
19.5
19.7
19.4
19.0
S.D.
0.9
1.3
1.1
0.8
0.9
1.0
0.9
3.2
0.8
0.6
0.9
0.7
0.5
1.2
(N)
22
65
61
17
15
11
8
7
10
10
16
26
12
280
Ear
Mean
16.2
15.8
15.2
16.3
15.7
15.5
14.6
15.2
15.9
16.6
17.1
16.9
16.9
15.9
Length
S.D.
1.3
1.1
1.2
1.3
0.9
1.4
0.7
1.3
1.1
0.6
0.8
0.8
0.9
1.3
(N)
22
65
61
17
15
11
8
7
10
10
16
26
12
280
-------
TABLE 6.6. HEMATOLOGICAL VALUES FOR Microtus ochrogaster COLLECTED NEAR
COLSTRIP.
July
1974 to
August 1975
•
Hematocrit, (%)
Summer
Winter
Hemoglobin, (g%)
Summer
Winter
Plasma protein, (g%)
Summer
Winter
N
20
13
13
6
20
13
Mean
49.80
52.23
15.42
17.20
6.22
5.97
S.D.
4.42
4.90
1.26
1.18
0.44
0.43
t
1.65
2.97
1.67
P
>0.05
<0.005
>0.05
TABLE 6.7. HEMATOLOGICAL VALUES FOR
COLSTRIP- July 1974 to
Peromysus
August 1975
maniculatus
COLLECTED NEAR
Hematocrit, (%)
Summer females
Winter females
Summer males
Winter males
All females
All males
Hemoglobin, (g%)
Summer females
Winter females
Summer males
Winter males
All females
All males
Plasma Protein, (g%)
Summer females
Winter females
Summer males
Winter males
All females
All males
N
60
18
114
25
78
139
37
5
75
6
42
81
43
18
86
22
61
108
Mean
48.48
50.06
49.88
52.24
48.85
50.30
15.85
14.45
15.87
15.29
15.68
15.82
6.39
6.03
6.39
6.17
6.28
6.35
S.D.
3.44
5.58
4.42
3.07
4.05
4.30
1.31
2.64
1.40
0.46
1.55
1.36
0.75
0.60
0.56
0.46
0.68
0.52
t
1. 14
--
3.19
--
2.50
1.17
--
2.32
--
0.50
1.98
--
1.90
--
0.70
P
>0.10
--
<0.005
--
<0.01
>0.10
--
<0.025
--
>0.30
<0.05
--
<0.05
--
>0.20
229
-------
TABLE 6.8. MEAN CORPUSCULAR HEMOGLOBIN CONCENTRATION IN RODENTS COLLECTED
NEAR COLSTRIP. July 1974 to August 1975.
Species and Season MCHC1
Microtus ochrogaster
Summer 31.0
Winter 32.9
Peromyscus mam'culatus
Summer females 32.7
Winter females 28.9
Summer males 31.8
Winter males 29.3
MCHCi = 100 (mean hemoglobin concentration), ted
mean hematocnt K
TABLE 6.9. HEMATOLOGICAL VALUES FOR Microtus ochrogaster COLLECTED NEAR
COLSTRIP. April
to September 1976.
Function
Sex
Hematocrit (%) M
F
Hemoglobin (g
%) M
F
Plasma protein (g %) M
F
N
11
12
11
10
11
12
Mean SD t P
43:73 z:?6 2-54 <0-02
15l9 0.77 2'45 <°-05
I'll ?'«n J-IS <0.40, NS
O . O / 1 . o U
TABLE 6.10.
HEMATOLOGICAL VALUE
COLSTRIP. April to
FOR Peromyscus maniculatus COLLECTED NEAR
September 1976.
Function
Hematocrit (%)
Hemoglobin (g
Plasma protein
Sex
M
F
%) M
F
(g %) M
F
N
93
49
79
37
106
55
Mean SD t * P
48^2 3' 10 4'34 <0-001
1 c no I oc
leios \zl 3-19 <0-01
6.20 1.06
6.21 0.72 "" Nb
230
-------
TABLE 6.11. HEMATOLOGICAL VALUES OF Perognathus fasclatus COLLECTED NEAR
COLSTRIP. April
to September
1976.
Function
Hematocrit (%)
Hemoglobin (g %)
Plasma protein (g %)
N
4
5
4
Mean
55.75
17.78
5.48
SD
2.36
1.74
0.92
TABLE 6.12. MEAN CORPUSCULAR HEMOGLOBIN CONCENTRATION IN RODENTS COLLECTED
NEAR COLSTRIP. April to September 1976.
Species
Peromyscus mam'culatus
Microtus orchrogaster
Perognathus fasciatus
Sex MCHC1
M
F
M
F
M+F
33.6
33.4
32.8
32.6
31.9
100 (mean Hb concentration) , , , -,- , , , „ ,,
= TT—r -, as computed from Tables 9, 10, 11
m63n ncT.
231
-------
As in 1974-75, male deer mice showed higher hematocrit readings compared
with females (Table 6.10). Males also showed higher hemoglobin values, which
was not found in the 1974-75 data (Table 6.7). Plasma protein was identical in
both sexes. The same values for P. fasciatus are tabulated in Table 6.11.
Although the sample size is too small for adequate comparison, hematocrit and
hemoglobin are a little larger than for either the deer mouse or the prairie
vole, while plasma protein is somewhat smaller.
The mean corpuscular hemoglobin concentration of voles and mice in 1976
was the same for both sexes (Table 6.12) and was 4-5% higher in 1976 than for
summer voles and mice in 1974-75.
Weights of several organs are presented in Table 6.13. In addition to
their basic reference value, these data provide indications of functional state
as well as annual cycle and life cycle trends and adjustments to environmental
factors. For example, greater liver and kidney size in females is probably
related to the increased metabolic demands imposed by reproduction. Mean liver
weight in 13 lactating females was 1567.6 mg (S.D. = 277.5) and in 9 non-
lactating females also collected in summer it was 1213.3 mg (S.D. = 257.2).
The difference was highly significant (t = 3.08, P < 0.005). Mean kidney
weight was also larger in these same lactating females, 363.3 mg vs. 233.1 mg
(t = 1.93, P < 0.05).
The significance of sexual dimorphism in adrenal gland size is unclear,
but it is a condition common to many eutherians (Christian, 1952; Chester-
Jones, 1957). As in this study, McKeever (1964b) found that female P_. mani-
culatus had larger adrenals than males. He showed that there was little
seasonal change in gland size, indicating that the difference is independent of
sex hormone levels, unlike the situation in M. montanus (McKeever, 1959).
McKeever (1964b) also found adrenal size to be independent of population
density, in agreement with Bendell (1959) but contrary to the findings of
Christian (1959) on P. leucopus.
POPULATION BIOLOGY
Five species of mice were captured in live traps: the deer mouse (Pero-
myscus maniculatus), prairie vole (Microtus ochrogaster), Wyoming pocket mouse
(Perognathus fasciatus), northern grasshopper mouse (Onychomys leucogaster),
and harvest mouse (Reithrodontomys megalotis). A few immature cottontails
(Sylvilagus auduboni) were captured in fall or early winter. Occasionally a
thirteen-lined ground squirrel (Spermophi1 us tridecemlineatus) or least chip-
munk (Eutamias minimus), both diurnal species, were taken if traps were opened
slightly before dusk.
This investigation is aimed primarily at nocturnal mice because of their
abundance and diversity. Of the species captured, P. maniculatus formed an
overwhelming portion of the catch. Members of this species were taken in every
trapping session in which animals were captured. They accounted for 77.4% (721
of 931) of the individuals captured in the period and 76.4% (220 of 288) for the
period April to June 1976. Furthermore, they represented 79.8% (1178 of 1476)
of the total captures made at the grid sites. At the end of September 1976 this
232
-------
TABLE 6.13. ORGAN WEIGHTS OF ADULT Peromyscus maniculatus COLLECTED NEAR
COLSTRIP. July 1974
to August
1975.1
Heart
Femal es
Males
Lungs
Females
Males
Liver
Females
Males
Spleen
Females
Males
Kidneys
Females
Males
Adrenal s
Females
Males
N
28
47
15
21
26
45
26
46
33
42
34
43
Mean
159.9
160.7
267.6
292.7
1427.0
1190.1
122.0
84.3
345.6
315.3
14.32
10.47
S.D.
28.5
27.3
65.9
59.7
317.6
271.8
151.4
48.7
51.5
37. 1
5.84
4.42
t
0.11
1.17
3.19
1.24
2.85
3.21
P
>0.45
---
>0.10
—
<0.005
—
<0.10
<0.005
—
<0.005
1mi11igrams
233
-------
figure was 71.8%, including recaptures of marked animals (Tables 6.14 and
6.15).
A capture summary of animals by species, sex and trapping location shows
that the five species were distributed in about the same proportions through-
out the study area, the Kluver East grid being an exception. At that location
no R. mega!otis were captured but 68.8% (11 of 16.) of the 0. leucogaster indi-
viduals taken in the 1974-5 study were captured there (Table 6.16).
Table 6.16 includes only individuals whose sex was determined. These
data indicate that in all species except P. maniculatus (Table 6.17) the sex
ratios were not statistically different. The sex ratio of P. maniculatus
during the spring and summer was similar to that found in 1974-75 at the same
site. However, 3-1/2 times as many male P. fasicatus were trapped as females.
During 1974-75, 59.3% (210 of 354) of P. maniculatus captured on grids
were males. Recapture rate was 58.6% in favor of males (268 of 457). There
was no indication of differental survival as seen in recapture tendencies
(Figure 6.7) or in overwinter survival (Table 6.18). Collectively, this
suggests that more males were trapped because more were present. The rela-
tively low percentage of females suggests also that the £. maniculatus popu-
lation was relatively low during the time of this study.
Trapping success is summarized in Table 6.19. Overall success never ex-
ceeded 10% in a given month. Having two traps per station undoubtedly contri-
buted to the relatively low catch per trap; still, there is every indication
that mouse populations were low. During a population high, for example, trap-
ping success can be at least as great as 44% for P. maniculatus (Hoffman,
1955).
There was considerable seasonal variation in the relative abundance of
species captured on the various grids (Figure 6.8). P. maniculatus was in-
variably the most abundant species, comprising from 52 to 95% of the total
monthly catch (Figure 6.8). 0. leucogaster and £. fasciatus are hibernators
and their absence from samples during winter was expected. R. mega!otis is
probably a hibernator also. However, one was taken in January 1975 and none
thereafter. A sharp decrease in the catch of M. ochrogaster occurred after
October 1974. Prior to January 1975 voles comprised 13 to 33% of the catch
(Lewis and Morton, 1975). This decrease in numbers of M. ochrogaster is unex-
plained but it could be part of a normal population cycle. Density-dependent
cycles occur with a periodicity of about four to five years (Krebs, 1966;
Batzli and Pitelka, 1970; Christian, 1971; Krebs et al_. , 1973) in microtine
rodents. A period of unusually severe weather coincided with the onset of
decreased catch of M. ochrogaster and R. megalotis. Winter temperatures were
generally mild in comparison to long-term averages through most of January
(Figure 6.9). From late January and through early March, however, temperatures
were at the seasonal low or oscillating enough to cause a daily freeze-thaw
cycle. Consequently, in four attempts at trapping in February and early March,
not a single animal was captured. The presence of fresh tracks in the snow at
trap entrances indicated that mice were abroad and active, but trap mechanisms
were invariably frozen and inoperative. Poorly functioning traps also affected
trapping success during periods of low temperature in November and December.
234
-------
90
80
70
60
"o
I 5°
°>
•o
- 40
O
30
20
E3 Males (N = I86)
g] Females (N = I30)
345
Times Captured
Figure 6.7. Recapture frequencies of Peromyscus maniculatus captured on
grids near Colstrip, July 1975 to July 1975.
235
-------
IND
CO
CT>
o
O
0>
o
V.
0>
0.
10
0
IO
0
EO
10
0
40
30
20
90
0
100
90
80
70
60
50
40
30
20
10
0
Onychomys leucogaster
r
Reithrodontomys megalotis "I
Perognathus fasciatus
Microtus ochrogaster
Peromyscus maniculatus
ASOND J FMA
Figure 6.8. Seasonal change in relative numbers of mice captured on grids near Colstrip.
-------
JVi
CO
Figure 6.9. Mean daily high and low ambient temperatures summarized at 5-day intervals (shaded area) and
monthly precipitation (histograms) from July 1974 through August 1975. Long-term monthly
averages are shown by solid symbols. Data recorded at Miles City, Montana, airport, which is
about 75 km from study sites and provided by Miles City Flight Service.
-------
TABLE 6.14. TOTAL AND RELATIVE NUMBERS OF MICE CAPTURED IN COLSTRIP STUDY,
July 10, 1974 to August 12, 1975.
Gri
Individuals
Peromyscus
maniculatus
Microtus
ochrogaster
Perognathus
fasciatus
Onychomys
leucogaster
Reithrodontomys
megalotis
TOTALS
354
70
26
12
12
474
ds
Recaptures
457
63
14
13
1
548
Collect!
367
63
11
4
9
457
Total
ons captures
1178
196
51
29
22
1476
% of total
captures
79.8
13.3
3.4
2.0
1.5
100.0
TABLE 6.15. TOTAL
12 to
AND RELATIVE NUMBERS
September 10, 1976.
OF MICE
CAPTURED NEAR
COLSTRIP, April
Grids
Indi
Peromyscus
maniculatus
Microtus
ochrogaster
Perognathus
fasciatus
Onychomys
1 eucogaster
Reithrodontomys
megalotis
TOTALS
vidual
38
13
14
0
1
66
Recaptures
127
49
18
0
1
195
Collect!
182
31
6
0
3
222
Total
ons captures
347
93
38
0
5
483
% of total
captures
71.8
19.3
7.9
*
0
1.0
100.0
238
-------
TABLE 6.16. SUMMARY OF CAPTURE RECORDS OF MICE ACCORDING TO SPECIES, SEX, AND TRAPPING LOCATIONS DURING
1974-75.
Species
Peromyscus maniculatus
Mai es
Females
Microtus ochrogaster
Males
Females
ro , . „
co Perognatnus Tasciat.us
Males
Females
Onychomys 1 eucogaster
Mai es
Females
Reithrodontomys megalotis
Males
Females
McRae
XI
79
57
20
14
4
11
0
0
2
3
2R
121
94
18
21
4
6
0
0
0
1
Kluver North
I
40
20
4
6
6
2
0
1
0
4
R
61
26
3
4
1
3
0
0
0
0
Kluver East
I
45
44
5
4
0
1
5
6
0
0
R
54
51
4
1
0
0
6
7
0
0
Kluver West
I
26
15
1
1
1
0
0
0
0
1
R
13
11
0
0
0
0
0
0
0
0
Garfield's
I
20
8
9
6
0
1
0
0
0
2
R
19
7
5
7
0
0
0
0
0
0
Col lections
I
220
142
33
30
4
6
3
1
6
3
1Number of individuals
2Number of recaptures
-------
TABLE 6.17. SEX DISTRIBUTION AND SEX RATIOS OF ALL MICE CAPTURED DURING
1974-75.
Species
Peromyscus maniculatus
Microtus ochrogaster
Perognathus fasciatus
Qnychomys leucogaster
Reithrodontomys megalotis
Males1
430
72
15
8
8
Females1
286
61
21
8
13
Ratio
1.50:1
1.18:1
0.7:1
1.00:1
0.62:1
P
<0.005
NS
NS
NS
NS
1Numbers of recaptures not included
TABLE 6.18. OVERWINTER SURVIVAL OF Peromyscus maniculatus.
Age
Period of
first capture Males
Recaptured
March-August 1975 Survival
Females Males Females %
Adults
July-August 74 31
20
9
September-
October 74
November-
December 74
January 75
TOTAL
68
Immatures July-December 74 24
23
16
3
_2
44
16
2
_4
17
0
_2
7
16.7
19.4
16.7
60.0
21.4
7.5
240
-------
TABLE 6.19. SEASONAL CHANGES IN SPECIES COMPOSITION AND TRAPPING SUCCESS ON COLSTRIP GRID.1
ro
1974
Species
Peromyscus maniculatus
Microtus ochrogaster
Perognathus fasciatus
Onychomys 1 eucogaster
Rei throdontomys megalotis
Total mice
Trap nights
% success, all mice
% success, Peromyscus only
July
51
13
0
0
0
64
1452
4.4
3.5
Aug
130
23
2
8
0
163
3388
4.8
3.8
Sept
95
19
19
2
8
143
1452
9.8
6.5
Oct
76
21
11
1
3
112
1452
7.7
5.2
Nov
33
11
0
0
1
45
968
4.6
3.4
Dec
36
14
0
0
1
50
968
5.2
3.7
Jan
31
16
0
0
1
48
484
9.9
6.4
Mar
28
2
0
1
0
31
720
4.3
3.9
Aor
98
10
0
2
0
110
1210
9.1
8.1
1975
May
76
2
5
4
0
87
1210
7.2
6.3
1976
June July May 19 Jun 10 Jun 30 Jul 21 Aug 1
77
1
1
6
0
85
1210
7.0
6.4
80 30 33 29 18 25
10 9 10 17 4
2 8 11 7 2 1
100000
002000
84 38 55 46 37 30
2178 --2 --2 — 2 -2 -z
3.9 5.2 8.3 6.6 5.4 4.2
3.7 4.1 5.0 4.2 2.6 3.5
Sep 1
30
6
3
0
0
39
2
5.5
4.2
'Total numbers of animals.
Approximately 720 each session.
-------
Other investigators have reported similar difficulties (Gunderson, 1950;
MacKay, 1962; Bergstedt, 1965).
Only a few mice of any species except P_. maniculatus were captured during
a given trapping session. In such cases there is negligible value in applying
procedures or models for predicting population size; the best estimate of pop-
ulation size is the minimum number of animals present, that is, the number
actually captured during a trapping session. This number, the minimum esti-
mate, is shown for all grid trapping efforts (Tables 6.20 - 6.23) along with an
estimate of population size for £. maniculatus by application of the method of
Jolly (1965). The Jolly stochastic model was the best of four methods tested
for predicting population size (French et a_L , 1971), but it may underestimate
the population (O'Farrell et a]_. , 1975). Only at Kluver North and McRae's were
sufficient capture records obtained to predict a population substantially
above that of the minimum estimate.
The similarity of Jolly's estimates to minimum estimates in many of the
trapping sessions indicates that nearly all P_. maniculatus present on the grids
were being captured and supports the conclusion from data on trapping success
and sex ratios that the population was low. The same might be inferred
regarding the numbers of other rodent species, but the minimum estimates are
suspect for several reasons. First, rolled oats may not be an effective bait
for primarily insectivorous species such as 0. leucogaster; even £. maniculatus
probably prefers insects when they are readily available (Williams, 1959;
Brown, 1966; Kritzman, 1974). In fact seasonal changes in acceptance of bait
by rodents may influence trapping success more than population density (Fitch,
1954). Second, weather conditions were variable. The unquantifiable effect of
frozen trap mechanism has been mentioned, but other factors such as rainfall
can be implicated. For example, only 38 captures of P_. fasciatus were made
during the entire study but 14 (37%) of these occurred on one rainy night in
September on the McRae grid (Table 6.20). Weather has also been identified as
affecting the response of rodents to traps (Getz, 1961, 1968; Gentry et. al_. ,
1966; Weiner and Smith, 1972; Gurnell, 1976). The suitability of sage-
grassland as rodent habitat is also questionable. In running collection
traplines in 1975, it was noted that the tops of steep little knolls with
outcrops of rocks such as those at Hay Coulee were comparatively more
productive for trapping P. maniculatus, although in 1976 flat areas were also
highly productive. Microtus runways were noted in numbers in old hay fields,
creek bottoms, and roadside ditches but not in grid areas. The best success in
catching M. ochrogaster on grids was at McRae's where a grassy swaTe bisected
the grid and at Garfield's where a swale flanked one side and where both Pony
Creek and Rosebud Creek bottoms were within a few hundred meters. Such grassy
areas probably effect a patchy distribution of M. ochrogaster because of their
suitability for runways rather than as food sources (Martin, 1956; Fleharty and
Olson, 1969; Grant and Morris, 1971).
HISTOLOGY
The study of histological cycles is designed to help interpret biological
change at the levels previously discussed (e.g., reproduction and development,
physiological condition, and population dynamics) and to aid in the identifi-
cation of direct and indirect effects of pollution. Tissues under investiga-
242
-------
TABLE 6.20. NUMBER OF MICE CAPTURED ON KLUVER NORTH GRID. MINIMUM ESTI-
MATE (ME) EQUALS TOTAL CATCH ON A GIVEN TRAPPING DATE AND
JOLLY ESTIMATE (JE) WITH STANDARD ERROR EQUALS POPULATION
SIZE PREDICTED BY JOLLY'S (1965) MODEL.
Date
Peromyscus
Microtus
Perognathus Onychomys
nianiculatus ochrogaster fasciatus
July 15 1974
August 2
August 13
August 23
September 13
October 8
November 5
December 5
March 20 1975
May 6
May 13
May 16
July 11
July 15
July 18
ME
5
9
9
4
16
12
11
15
15
14
11
11
5
3
3
JE
18.
16.
6.
23.
17.
21.
41.
33.
16.
12.
10.
9.
12.
--
9
7
7
1
5
4
2
8
2
3
0
0
0
-
9
5
3
6
5
7
23
17
4
3
3
4
-
-
SE
.0
.4
.7
.8
.4
.8
.5
.0
.4
.6
.2
.3
--
--
ME
0
1
0
0
4
1
1
6
0
1
1
0
0
0
0
ME
0
0
0
1
0
5
0
0
0
1
2
2
0
0
0
leucogaster
ME
0
3
0
0
0
0
0
0
1
0
0
0
0
0
0
Reithrodontomys
megalotis
ME
0
0
0
0
1
2
1
0
0
0
0
0
0
0
0
243
-------
TABLE 6.21. NUMBER OF MICE CAPTURED ON KLUVER EAST GRID. MINIMUM ESTIMATE
(ME) EQUALS TOTAL CATCH ON A GIVEN TRAPPING DATE AND JOLLY
ESTIMATE (JE) WITH STANDARD ERROR EQUALS POPULATION SIZE PRE-
DICTED BY JOLLY'S (1965) MODEL.
Date
Peromyscus
maniculatus
ME
July 23 1974
August 6
August 16
August 30
September 20
October 12
November 15
December 13
March 25 1975
May 28
May 29
June 3
June 6
July 25
July 30
August 1
1
1
1
3
8
1
3
13
8
7
2
8
23
1
1
1
1
1
1
7
0
1
1
0
3
JE
9.
14.
15.
16.
8.
15.
6.
8.
23.
20.
12.
24.
43.
10.
7
2
3
5
0
2
0
0
0
5
9
4
3
0
Microtus
Perognathus Onychomys
ochrogaster fasciatus leucogaster
SE
3.
4.
4.
5.
2.
11.
5.
2.
4.
4.
3.
13.
40.
--
3
5
4
2
8
5
8
8
8
9
8
4
5
-
ME
2
1
0
4
2
1
4
0
0
0
0
0
0
0
0
0
ME
0
0
0
1
0
0
0
0
0
0
0
0
0
0
0
0
ME
0
3
2
3
2
1
0
0
0
2
2
3
3
0
1
0
Reithrodontomys
megalotis
ME
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
244
-------
TABLE 6.22. NUMBER OF MICE CAPTURED ON KLUVER WEST GRID. MINIMUM ESTIMATE
(ME) EQUALS TOTAL CATCH ON A GIVEN TRAPPING DATE AND JOLLY
ESTIMATE (JE) WITH STANDARD ERROR EQUALS POPULATION SIZE PRE-
DICTED BY JOLLY'S (1965) MODEL.
Date Peromyscus Microtus Perognathus Onychomys Reithrodontomys
maniculatus ochrogaster fasciatus leucogaster megalotis
ME JE SE ME ME ME ME
July 15 1974 4 — ---0 0 0 0
August 23 — — 0 0 0 0
August 13 4 12.0 11.5 000 0
August 23 6 10.5 5.9 0 0 0 0
September 17 8 8.0 2.8 0 0 0 1
October 11 12 12.0 3.5 0 1 0 0
November 12 12 16.8 6.3 1 0 0 0
December 10 9 17.1 --- 1 0 0 0
May 20 1975 5 — -—0 0 0 0
245
-------
TABLE 6.23. NUMBER OF MICE CAPTURED ON MCRAE'S GRID. MINIMUM ESTIMATE
(ME) EQUALS TOTAL CATCH ON A GIVEN TRAPPING DATE AND JOLLY
ESTIMATE (JE) WITH STANDARD ERROR EQUALS POPULATION SIZE
PREDICTED BY JOLLY'S (1965) MODEL.
Date
Peromyscus
Mi
crotus
maniculatus ochrogaster
July 10 1974
July 28
August 9
August 20
September 10
September 27
October 1
October 22
December 3
January 25 '75
April 1
April 4
April 15
April 18
June 10
June 17
June 24
July 2
ME
11
14
16
21
28
18
13
23
9
20
17
21
21
28
23
16
18
14
JE
-•
20
17
21
28
27
24
28
26.
36.
24.
28.
28.
30.
33.
22.
25.
17.
--
.5
.3
.0
.0
.3
.0
.7
,7
2
,5
9
3
4
2
6
7
9
SE
--
6.
4.
4.
5.
6.
6.
7.
10.
10.
5.
6.
6.
6.
8.
5.
6.
--
• -
4
2
6
3
7
8
0
8
6
8
2
0
3
6
3
5
-
ME
2
6
7
7
3
7
5
8
7
10
2
4
2
1
1
0
0
0
Perognathus
fasciatus
ME
0
0
0
0
3
14
1
4
0
0
0
0
0
0
0
1
0
2
Onychomys
leucogaster
ME
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Reithrodontomys
mega! otis
ME
0
0
0
0
2
2
1
1
0
0
0
0
0
0
0
0
0
0
246
-------
tion can be categorized as indicators of stress (e.g., adrenal and thyroid
glands), indicators of immunological change (e.g., spleen), indicators of
toxicological stress (e.g., lung, liver, and kidney), and indicators of repro-
ductive function (e.g., gonads and accessory glands). Only the major repro-
ductive structures of immature and adult £. maniculatus and M. ochrogaster are
treated in this report. Portions of the testes and associated structures are
described synoptically in Tables 6.24 to 6.31; these are further described and
discussed in the associated text.
Seminiferous tubules
The body of the testis contains many pyramidal compartments which contain
one to several seminiferous tubules, each bounded by a thick tunica albuginea of
regularly arranged collagen fibers. Within these tubules the spermatozoa or
male germ cells are formed and are visible in the tubule walls along with
developing gametes and Sertoli cells, the major supporting component of the
seminiferous epithelium. A regular feature of the tubules, especially when they
are spermatogenetically inactive, is one-to-few round eosinophilic droplets the
size of mature primary spermatocytes. These occur in the tubular lumen or among
the layers of gametes lining its wall. They have been previously described and
are interpreted as degenerate gametocytes (e.g., Clarke and Kennedy, 1967).
Spermatogenic cells at various stages of development also line the tubule.
These are described in order of development: spermatogonia or gonia, primary
spermatocytes, secondary spermatocytes, spermatids, and spermatozoa. Sperma-
togonia multiply by mitosis. Frequent mitotic spermatogonia appear in the
seminiferous tubules. They contain chromosomes, are small in size, and line the
margins of the seminiferous tubule. However, these dividing gonia are distinct
from primary or secondary spermatocytes that undergo meiosis.
Interstitial tissue
The interstitial tissue or Cells of Leydig represent the endocrine compo-
nent of the testis. They are located in areas around the convoluted semin-
iferous tubules. They elaborate several steroid hormones, especially
testosterone, without which the seminiferous epithelium does not sustain
proliferation and differentiation of the germ cells. Testosterone also supports
the normal development of accessory glands of the reproductive tract, such as
the seminal vesicles. When active, Leydig cells are easily recognized. When
inactive, they cannot be distinguished from other connective tissue cells that
lie between the seminiferous tubules. Leydig cells classified on the basis of
cytoplasmic characteristics are as follows:
1. inactive: appear as other connective tissue cells, e.g., fibro-
blasts;
2. moderately active: round nucleus and cells, but with only a small
amount of cytoplasm; nucleus occupies most of the cell volume;
3. active: round nucleus and cells; cytoplasm more abundant and fre-
quently filled with secretory granules or small vesicles;
247
-------
TABLE 6.24. GENERAL SUMMARY -- IMMATURE TESTIS Peromyscus maniculatus.
Seminiferous Tubules
Month
July
Aug
Sept
Oct
Nov
-£> Dec
Jan
Mar
May
June
July
Aug
Year
1974
1974
1974
1974
1974
1974
1975
1975
1975
1975
1975
1975
Average Diameter1
(u ± SE) (n)
121.5 (1)
75.8 (3)
73.2 ± 10.6 (10)
56.4 ± 4.6 (10)
56.3 ± 3.2 (8)
72.0 ± 8.6 (6)
105.6 ± 8.2 (5)
152.0 (2)
139.2 ± 13.9 (4)
145.0 (3)
78.4 ± 7.6 (14)
73.6 ± 18.6 (6)
Most
Advanced
Stage2
Spermatozoa
Primaries (2)
Spermatozoa (1 )
Gonia (1 )
Primaries (5)
Secondaries (2)
Spermatozoa (2)
Primaries (9)
Spermatozoa (1 )
Primaries
Primaries (1)
Secondaries (4)
Spermatozoa (1)
Spermatozoa
Spermatozoa
Spermatozoa
Spermatozoa
Primaries (2)
Secondaries (1)
Spermatids (6)
Spermatozoa (5)
Primaries (4)
Spermatozoa (2)
Mitotic
Gonia
Many
Some
None (1)
Some (2)
Many (7)
None (1)
Many (9)
None (1)
Many (7)
None (4)
Some (2)
None (2)
Many (3)
Many
Some (1)
Many (3)
Many
None (8)
Some (3)
Many (2)
None (2)
Some (2)
Many (2)
First
Me i otic
Figures3
None
None (1)
Many (2)
None (6)
Many (4)
None (3)
Some (1 )
Many (6)
None (1)
Some (3)
Many (4)
Some (2)
Many (4)
None (4)
Many (1)
None
None (3)
Many (1)
None (1)
Some (1 )
Many (1)
None (4)
Some (3)
Many (6)
None (4)
Many (2)
Degenerating
Gametes
None
None (2)
Some (1)
None (3)
Some (7)
None (2)
Some (7)
Many (1)
None (2)
Some (7)
Many (2)
None (2)
Some (4)
None (4)
Some (1)
None
None
Some ( 1 )
None
None (6)
Some (5)
Many (3)
None (1)
Some (3)
Many (2)
Activity of
Leydig Cells
Mod. Active
Inactive (2)
Mod. Active (1)
Inactive (9)
Mod. Active (1)
Inactive
Inactive (6)
Mod. Active (2)
Inactive (4)
Mod. Active (1)
Active (1)
Active
Very Active
Active (3)
Very Active (1 }
Active (2)
Very Active (1)
Inactive (10)
Mod. Active (2)
Active (2)
Inactive (4)
Mod. Active (1)
Very Active (1)
'Microns ± standard error; number of samples in parenthesis.
2Primaries = primary spermatocytes; secondaries = secondary spermatocytes.
3Second Meiotic figures in immature testis are not included.
-------
TABLE 6.25. GENERAL SUMMARY — ADULT TESTIS Peromyscus maniculatus.1
ro
Seminiferous Tubules
Month
July
Aug
Sept
Oct
Nov
Dec
Jan
Mar
Apr
May
June
July
Aug
Year
1974
1974
1974
1974
1974
1974
1975
1975
1975
1975
1975
1975
1975
Average Diameter
(y ± SE) (n)
159.8 ± 6.1 (4)
148.5 ± 9.6 (5)
133.7 ± 9.4 (8)
90.8 ± 12.8 (8)
64.0 (1)
99.2 ± 8.9 (4)
102.0 (1)
157.0 (3)
163.2 ± 14.0 (7)
196.0 ± 18.0 (10)
162.7 ± 4.6 (18)
162.8 ± 5.8 (14)
162.9 ± 7.3 (10)
Most
Advanced
Stage
Spermatozoa
Spermatozoa
Primaries (1)
Spermatozoa (7)
Gonia (1)
Primaries (3)
Spermatozoa (4)
Primaries
Primaries (1 )
Spermatozoa (3)
Spermatozoa
Spermatozoa
Spermatozoa
Spermatozoa
Spermatozoa
Primaries (1 )
Spermatozoa(lS)
Spermatozoa
Mitotic
Gonia
Many
Many
None (1)
Some (1)
Many (6)
None (2)
Some (1 )
Many (5)
Many
None (1)
Some (3)
Many
None (1)
Some (1 )
Many (1)
None (2)
Some (2)
Many (3)
None (6)
Many (4)
Many
None (3)
Many (11)
None (2)
Many (8)
First
Meiotic
Figures
None
None (4)
Many (1)
None (1 )
Some ( 1 )
Many (6)
None (3)
Some (3)
Many (2)
Many
Many
Many
Many
None (6)
Some (1)
None (6)
Some (4)
None (10)
Some (1 )
Many (7)
None (8)
Some (2)
Many (4)
None (7)
Some (2)
Many (1)
Second
Meiotic
Figures
None
None
None
None
None
None
None
None
None
None (9)
Some (1)
None
None
None (9)
Many (1)
Degenerating
Gametes
None (3)
Some (1)
None (4)
Some (1)
None (4)
Some (4)
None (6)
Some (2)
None (3)
Some (1)
None
None
None
None (9)
Some (1 )
None (17)
Some (1)
None
None (9)
Some (1)
Activity of
Leydig Cells
Active (2)
Very Active (2)
Inactive (1)
Mod. Active (1)
Active (1)
Very Active (2)
Inactive (1)
Mod. Active (2)
Active (4)
Very Active (!)
Inactive (5)
Mod. Active (1)
Active (1)
Very Active (1)
Inactive
Active
Active
Active (2)
Very Active (1 )
Active (1)
Very Active (6)
Active
Mod. Active (1)
Active (7)
Very Active(lO)
Inactive (1)
Mod. Active (4)
Active (5)
Very Active (4)
Inactive (1)
Mod. Active (1)
Active (2)
Very Active (6)
'Explanations as in Table 6.24.
-------
TABLE 6.26. GENERAL SUMMARY -- TAIL OF EPIDIDYMIS OF IMMATURE Peromyscus maniculatus
Month
Aug
Mar
June
July
ro .
en Aug
Epithel ium
Year
1974
1975
1975
1975
1975
Type
Col1
Cub2
Col
Ps3
Cub
Col
Ps
Cub
Col
Cub
(1)
(2)
(2)
0)
(3)
(3)
(1)
(1)
0)
Height
(M)
10.5
18.0
19.5
10.5
17.8
(n)
(1)
(2)
(3)
(1)
(2)
Cilia
Many
Many
Many
Many
Some (1)
Diameter
(M) (n)
222.5
113.5
147.0
226.5
260.5
(D
(2)
(3)
(1)
(2)
Tubule
Contents
Sperm4
HD
2(2)
3(2)
HD
2(1)
Empty5 Other6
None
None
1(0)
None
HO)
None
None
Fluid (1) (0)
None
None
= simple columnar epithelium.
2Cub = simple cuboidal epithelium.
3Ps = pseudostratified columnar epithelium.
4lst number indicates number of animals with some sperm. Number in parentheses are those with sperm in
all tubules.
5lst number indicates number of animals with some empty tubules. Number in parentheses are the number
of mice with fully empty tubules.
6This column describes tubule contents other than sperm. The first number indicates number of animals
having some of the contents described, while that in parentheses indicates those having all of the
contents.
7Degenerating.
8Lumen contains granular material.
-------
TABLE 6.27 GENERAL SUMMARY -- HEAD OF EPIDIDYMIS OF IMMATURE Peromyscus maniculatus!
Month Year Type
Aug 1974 Col1
Sept 1974 Cub2
Col
Ps3
Oct 1974 Cub
Col
Ps
Nov 1974 Col
Ps
ro
— • Dec 1974 Col
Ps
Jan 1975 Ps
Mar 1975 Col
Ps
May 1975 Cub
Col
Ps
July 1975 Cub
Col
Ps
(\ug 1975 Col
Ps
(1)
(2)
(7)
(3)
(4)
(9)
(1)
(3)
(2)
(2)
(2)
(2)
(2)
(4)
(2)
(1)
(14)
(11)
(3)
(3)
Epithelium
Height
(y ± SE) (n)
15.5 (1)
15.5 ± 5.6 (9)
14.1 ± 0.8 (9)
14.6 (3)
19.0 (3)
18.0 (1)
22.8 (2)
20.8 ± 3.0 (4)
17.8 ± 1.0 (14)
22.0 (3)
Tubule
Cil
None
?
None
Many
1
Some
Many
None
Some
None
Many
Ma ny
Many
Many
None
Some
Many
Some
Many
la
(1)
(3)
(5)
(1)
(2)
(6)
(2)
(1)
(1)
(2)
(2)
(1)
(11)
(1)
(2)
Diameter
(y ± SE) (n)
50.5 (1)
45.0 ± 7.4 (9)
48.6 ± 6.8 (9)
42.5 (3)
59.0 (3)
71.0 (1)
108.0 (2)
108.8 ± 12.6 (4)
78.4 ± 11.6 (14)
91.6 (3)
Sperm''
0(0)
2(1)
0(0)
0(0)
0(0)
1(0)
2(1)
4(0)
3(0)
3(0)
Empty5
1(0)
8(4)
9(9)
3(3)
3(2)
1(0)
None
4(0)
14(8)
3(0)
Contents
Other6
Fibroblasts
Deg.7 lymphocytes
Epithelial cells
or lymphocytes
Granules8
Fluid
None
None
Granules
Granules
Deg. Sperm
Sloughed epith.
cells
None
Granules
Heterophils
None
(D
(1)
0)
(1)
(2)
(1)
(1)
(1)
(1)
(2)
(1)
(0)
(0)
(0)
I n \
\
-------
TABLE 6.28. GENERAL SUMMARY -- TAIL OF EPIDIDMYIS OF ADULT Peromyscus maniculatus.1
Month
Sept
Oct
Mar
Apr
ro
en
May
June
July
Aug
Year
1974
1974
1975
1975
1975
1975
1975
1975
Type
Cub (1)
Col (5)
Ps (3)
Col
Cub (1)
Col (1)
Cub (5)
Col (6)
Cub (3)
Col (5)
Cub (6)
Col (7)
Ps (1)
Cub (3)
Col (8)
Ps (4)
Cub (2)
Epithel ium
Height
(y ± SE) (n)
17.9 ± 0.7 (5)
15.8 (2)
11.0 (1)
12.8 ± 0.6 (6)
12.7 ± 0.7 (5)
13.4 ± 1.0 (7)
20.4 ± 2.1 (8)
13.3+1 .5 (5)
Tubule
Cilia
Many
Many
Many
Many
Many
Many
Many
Many
Diameter
(v ± SE) (n)
165.1 ± 12.1 (5)
200.5 (2)
113.5 (1)
219.5 ± 12.7 (6)
226.1 ± 7.0 (5)
261.5 ± 11.8 (7)
170.8 ± 9.5 (8)
192.6 ± 18.6 (5)
Sperm
5(5)
2(2)
KD
6(6)
5(2)
7(6)
8(1)
5(4)
Contents
Empty Other
None None
None None
None None
None None
3(0) Fluid (1) (0)
1(0) None
7(0) None
1(0) None
'Abbreviations and explanations as in Table 6.26.
-------
TABLE 6.29. GENERAL SUMMARY -- HEAD OF EPIDIDYMIS OF ADULT Peromyscus maniculatus,1
Month
July
Sept
Oct
Dec
ro
en
oo
Mar
Apr
May
June
July
Aug
Year
1974
1974
1974
1974
1975
1975
1975
1975
1975
1975
Type
Col
Ps
Col
Ps
Col
Ps
Col
Ps
Col
Col
Ps
Cub
Col
Ps
Cub
Col
Ps
Col
Ps
Col
Ps
(5)
(6)
(2)
(8)
(6)
(4)
(1)
(6)
(4)
(1)
(18)
(17)
(7)
(7)
(6)
(5)
Epithel ium
Height
(y ± SE) (n)
20.5 ± 1.8 (1)
22.8 ± 2.1 (7)
19.1 i 1.8 (8)
24.5 (1)
23.5 (1)
18.8 ± 3.9 (6)
21.5 ± 0.3 (6)
22.6 ± 1.5 (18)
25.4 i 0.8 (7)
23.9 ± 1.3 (6)
Tubule
Cilia
Many
Many
? (1)
Many (7)
Many
Many
Many
Many
Many
Many
Many
Diameter
(y i SE) (n)
122.5 ± 3.4 (1)
80.6 i 7.6 (7)
66.3 i 8.0 (8)
Contents
Sperm
1(0)
5(2)
2(1)
Empty
1(0)
5(0)
7(4)
Other
None
Deg. sperm
Granules
Monocytes
Deg. sperm
(2)
(1) (0)
(1)
(1)
Prim. spermatocytes(l ) /n^
77.0 (1)
123.0 (1)
109.3 ± 10.6 (6)
116.2 t 7.5 (6)
124.4 i 6.0 (18)
108.9 ± 4.0 (7)
123.8 ± 27.5 (6)
1(0)
KD
6(2)
6(2)
17(7)
7(0)
6(1)
1(0)
None
4(0)
4(0)
19(0)
7(0)
5(0)
Pycnotic nuclei
debris
Fluid
None
None
None
None
Deg. sperm
Monocytes
Fluid
Cell debris
None
None
-
& *• '
(1)
(2)
(1)
(]) (0)
(2) (0)
(1)
Abbreviations and explanations as in Table 6.26.
-------
TABLE 6.30. GENERAL SUMMARY -- HEAD OF THE EPIDIDYMIS OF ADULT Microtus ochrogaster!
ro
en
Month
Aug
Nov
Dec
Aug
Aug
Sept
Mar
Apr
May
Aug
Year
1974
1974
1974
1975
1974
1974
1975
1975
1975
1975
Type
Ps
Col
Ps
Col
Ps
Col
Ps
Col
Ps
Col
Ps
Cub
Col
Ps
Col
Ps
Ps
(1)
(1)
(2)
(3)
(1)
(1)
(2)
(2)
(1)
(3)
(3)
(2)
(2)
Epithelium
Height
(y ± SE) (n)
13-.0 (1)
17.0 (1)
20.5 (2)
20.5 (2)
25.3 (3)
27.0 (1)
26.0 (2)
18.7 (3)
20.2 (2)
30.0 (2)
Tubule
Cilia Diameter
(y ± SE)
Immature Microtus
Many 47.0
Many 67.0
Many 63.5
None (1) 74.8
Many (1)
Adult Microtus
Many 112.5
Many 151.0
Many 95.5
Many 85.2
Some (1) 107.0
Many (1)
Some (1) 120.2
Many (1)
(n)
(1)
(1)
(2)
(2)
(3)
(1)
(2)
(3)
(2)
(2)
Sperm
None
1(0)
None
None
3(1)
KD
2(1)
2(2)
2(0)
2(1)
Contents
Empty
KD
None
2(0)
2(0)
2(0)
None
1(0)
KD
2(0)
1(0)
Other
None
Fluid
Gametes
Deg sperm
Debris
Deg sperm
Fluid
Monocytes
Heterophil
Debris
Fluid
None
None
None
None
None
(1)
(1)
(1)
(1)
(1)
(1)
(1)
(1)
(1)
(0)
CO)
(0)
(0)
'Abbreviations and explanations as in Table 6.26.
-------
TABLE 6.31. GENERAL SUMMARY — TAIL OF EPIDIDYMIS Microtus ochrogaster.1
Epi the! ium
Month Year Type Height
Dec 1974 Ps
cn
01 Mar 1975 Col
Ps
Apr 1975 Col
Ps
May 1975 Col
Ps
June 1975 Col
Ps
(M)
22.0
(1) 12.0
CD
(2) 24.8
(2)
(1) 18.5
(1)
(1) 18.0
CD
(n)
CD
(1)
(2)
(1)
(1)
Tubule
Cilia Diameter Contents
(u) (n) Sperm Empty Other
Immature Microtus
Many 206.5 (1) None None Granules (1)
Cell debris (1)
Heterophils (1)
Adult Microtus
Some 244.0 (1) 1(1) None None
Many 198.0 (2) 2(2) None None
Many 298.0 (1) 1(0) None Red Droplets (1)
None 263.5 (1) 1(1) None None
(1)
(0)
Abbreviations and explanations as in Table 6.26.
-------
4. very active: cell round, but larger than active or moderately active
cells; cytoplasm very abundant and generally filled with secretory
granules or small vesicles.
Epididyim's
The epididymis is an organ comprised of a single duct formed by the fusion
of many smaller excretory ducts from the testis. It stores spermatazoa, and is
believed to enable the sperm to develop the ability to fertilize ova and to
move normally. The epididymis is thus seen as an organ that is critical to the
final stage of sperm development. The epididymis is comprised of head, body
and tail. The head and tail differ markedly in histological structure. They
conform to descriptions of two regions of the epididymis of field voles (Lecyk,
1962) and are thought to be equivalent to these regions.
The tubules in the head region are relatively small and round. They are
lined by a simple high columnar to pseudostratified columnar epithelium rein-
forced by a thin capsule of connective tissue. Epithelial cells in reproduct-
ively active males have basal oval nuclei and much apical cytoplasm. The
latter is filled with secretory granules and sometimes contains a single large
unstained apical vacuole as well. The cells generally feature many long stero-
cilia. The lumen and tubule are round and regular in outline, and the tubules
are separated from each other by variable amounts of vascular connective
tissue. In a nonreproductive state, the tubules shrink in size, the simple
epithelium diminishes in height, and the cells are densely crowded. The tubule
lumen is reduced and often disappears entirely. The cilia are difficult to
locate and are completely absent in many cases.
The tubules in the tail region are very large, irregular in shape, and are
generally filled with sperm. The epithelium is simple low columnar to
cuboidal. Cell nuclei are centrally located and the cytoplasm is scanty.
Cilia are difficult to discern.
Degenerating sperm or degenerate cells resembling primary gametocytes are
sometimes observed in the epididymal tubules. Occasionally, the tissue
contains focal infiltrations of lymphocytes and sometimes heterophilic poly-
morphonuclear leucocytes in the connective tissue between the tubules.
Epithelial height and tubule diameter of adult mice in any gjven month
exceed those of immatures in the head but not the tail of the epididymis. Fur-
thermore, seasonal changes in epithelial height and tubule diameter are not
always in the same direction. Because diameter changes in tubule lumen from
month to month are larger than changes in epithelial height, diameter may be a
more sensitive measure of functional changes throughout the year.
BIRDS
INTRODUCTION
Birds'are the most visible vertebrates of the grassland ecosystem under
study during the period of spring to late fall. The dominant species are the
Western Meadowlark (Sturnella neglecta), Mourning Dove (Zenaidura macroura),
256
-------
Lark Bunting (Calamospiza melanocorys), Vesper Sparrow (Pooecetes gramineus),
and Lark Sparrow (Chondestes grammacus). The Mourning Dove occurs near riparian
habitats, where drinking water and the presence of trees for nesting are found,
but the other species can be found in a variety of grassland or sage-grassland
areas. Within these habitats, only the meadowlark seems to be well distributed.
The others often show discontinuity in their distribution.
Birds are potentially important indicator species because they are rela-
tively long-lived secondary consumers. Such forms are highly vulnerable when
ecosystem damage occurs (e.g., Stickel, 1975). Furthermore, they are unlikely
to show the acute effects of pollution and most likely to suddenly and belatedly
exhibit severe damage when pollutants have accumulated to toxic levels in the
organ-systems (Stickel, 1975). Their long lives enhance potential for accum-
ulation of pollutants. Even low-level, non-lethal accumulations of pollutants
which do not directly debilitate may have important effects on later generations
of birds and mammals (summarized by Stickel, 1975 and Truhaut, 1975).
Birds are day-active and typically roost above ground, in contrast to the
nocturnal fossorial mammals that are under study. The two groups will thus have
very different pollutant exposure histories. Investigation of the effects of
diurnal variation in power plant emissions is one of special interest and
potential importance. Mourning Doves, for example, feed most intensively during
the first few daylight hours and again late in the day. Clearly, this may
influence both their rate of exposure and susceptibility to air pollutants. The
species of birds under study employ different feeding strategies, namely,
granivory (dove), insectivory and carnivory (meadowlark), or both (bunting and
sparrows). If different food categories are differentially affected by pollu-
tion damage, then it should be possible to determine the effects from assay of
physiological conditions. For example, sulfur dioxide reduces pollination and
pollen germination and therefore the quality of seeds (Garber, 1967). Of the
birds in question such an effect would be expected to affect seed-eaters select-
ively. Examination of these species at histological levels and integration of
tissue and gross data may reveal diet-related effects of stack emissions and
hence the dietary group which is most useful as a bioindicator of pollution
stress.
Members of higher trophic levels, such as birds, may accumulate pollutants
for long periods before overt responses are seen. As indicated in a recent Oak
Ridge Workshop (Johnson, 1976), there is a general need for information
concerning the sublethal and chronic, cumulative effects of pollutants. The
"threshold" exposure rates that might be expected to produce various biological
effects (Lewis, Glass, and Lefohn, 1975; p. 142) on organisms are not yet known,
especially in areas such as Col strip where the impacts of pollutants are
continuous, but small. It is thus desirable to determine which tissues
selectively accumulate pollutants (Truhaut, 1975). By identifying avian
tissues that selectively accumulate or react strongly to pollutants, it may be
possible to anticipate when or with how much exposure a dramatic and sudden
effect will be overtly manifested in a species.
Some birds are sensitive indicators of pollution (Lewis, Glass, and Lefohn,
1975, Ch. 8). For example, pigeons living in highly industrialized areas of
Japan develop lung pathologies when aerial pollution levels are only one-tenth
257
-------
the value of those necessary to produce analogous pathologies in human lungs.
Birds appear to be particularly useful for measuring levels of air quality
because of the design of their respiratory system and their broad and regular
dispersion throughout the ecosystem. In contrast to bioindicating organisms of
lower trophic levels, birds provide a mechanism of separately assessing the
effects of aerial and dietary pollutants. The lungs and respiratory tract are
particularly valuable for monitoring air pollution. In contrast, ingested
pollutants probably affect the liver and perhaps the kidney selectively.
Intensive work is proceeding on the avian lung. This organism is struct-
urally analogous to a high volume sampler involved in aerial pollutant scaveng-
ing. It is a primary target for air-borne particulates because the nasal pass-
ages have little ability to remove them from inhaled air and because flow across
the respiratory surface is in only one direction (Bretz and Schmidt-Nielsen,
1971). As the studies of Japanese pigeons suggest, the avian lung is damaged by
rather small amounts of toxicants from coal-fired power plants. It should be
possible to determine even low emission rates and types from stacks of this kind
by periodically examining the lungs of birds in impacted areas. Coal dust,
silica, and the fugitive dust of agricultural activities are all somewhat
different in appearance and likely different with respect to the inflammatory
responses which they produce in avian respiratory tissues.
The avian Tung also seems to offer unusual potential for use in pollution
gradient analysis. Stack emissions settle out at distances from power sources
which vary with the type of pollutant. Heavy particles settle in the immediate
vicinity of the plant. However, particles dispersed in the form of aerosols are
transported great distances from the source and carry toxic gases and trace
metals. This, coupled with the broad and fairly regular dispersion of birds in
the grassland ecosystem, should permit (1) distinguishing the effects of
pollution from the effects of other environmental stresses, and (2) through
pollution gradient analysis, evaluating the effects of different concentrations
of stack emissions acting on birds over an extended period.
POPULATION BIOLOGY
Census data can yield information on the relative spatiotemporal abundance
of bird species. When a census is repeated at regular intervals, changes in
species frequencies and dispersion patterns can be deduced and these can be
related to changes in concurrently measured environmental parameters.„
By application of the method described below, the investigators hope to
relate changes in easily measurable descriptive parameters such as species di-
versity, equitability, and species richness to changes in functional relation-
ships between species and their habitats. Of particular interest are changes
attributable to the operation of the coal-fired power plants at Colstrip. This
report summarizes the baseline data and suggests a format for the analysis and
interpretation of future census data.
258
-------
Methods
The method employed is patterned after the North American Breeding Bird
Survey (Robbins and Van Velzen, 1970). In application, the observer starts
one-half hour before local sunrise and makes 60 three-minute stops at 0.5 mi
(0.8 km) intervals along a predetermined route (Figure 6.10). At each stop the
number of birds of each species seen in a 400 m radius and heard, regardless of
distance, is recorded. Since bird species have differential individual
detectability, the census data provide no information on absolute abundance
(Emlen, 1971). However, it should be possible to detect changes in species
relative abundance patterns by comparing data obtained during a baseline period
with those obtained using the same census technique at other times.
The census route was selected to provide sampling stations at various
distances from the Colstrip power plants along an anticipated gradient of pol-
lution impact. Individual stations span a wide range of habitats including
open grassland, streams, rolling hills with ponderosa pine and juniper
coverage, cliffs and habitats affected by a wide range of human impacts
including urban activities, railroad right-of-way, farming, ranching, and
mining.
The 1975 breeding season will be considered the baseline period for pres-
ent purposes since the power plants were not operational at that time. The
route in Figure 6.10 was censused on nine dates during the baseline period (May
6, May 23, June 11, June 30, July 14, July 28, August 11, August 26, and
September 8).
s
Species diversity was measured by the Shannon-Weaver function (H'=-l p.
i=l
Iog2p.) (Shannon and Weaver, 1949), species richness (S) was measured by the
number of species present, and equitability (E) was measured by H'/log2S
(Pielou, 1977). To inspect overall temporal trends in these parameters, total
H1, total S, and total E were calculated from lumped census data for all sites
on each sampling date. Spatial and site specific trends were inspected by
calculating total across season H1 , S, and E from lumped data from all sampling
dates for each site.
Results and Discussion
Evaluation of pollution impact depends heavily upon a comparison of future
species frequencies and dispersion patterns with baseline patterns established
prior to power plant operation. It is assumed that the parameters measured are
not in the process of reaction to the environmental influences extrinsic to the
ecosystem under scrutiny. Limited data from the U.S. IBP Grassland Biome
program indicate that grassland bird species diversity, equitability, and
species richness as measured in the present study do not vary substantially
between years, but generally decrease along a gradient of decreasing primary
production (Wiens, 1974, Table 6.3). H', E, and S are apparently buffered from
response to normally encountered environmental variations but may respond to
changes in productivity of the ecosystem brought about by chronic air
pollution. If so, the roadside bird census may provide an inexpensive method
of indexing long-term ecosystem response to chronic pollution.
259
-------
I06°45
T. 2 N.
R. 43 E.
T. 2 N.
I06°45'
R 42 E
6 !06°30
R. 43 E.
Figure 6.10. Map of the Rosebud-Col strip roadside census route.
260
-------
Temporal trends in species frequencies are presented in Table 6.32.
Though 63 species were recorded on the census route during 1975, most were
relatively rare. Meadowlarks were clearly predominant and the six most common
species contributed nearly 62% of total bird abundance. Diversity (H1) peaks
in late May, decreases until late July, and then remains fairly stable into
September. Seasonal trends in E and S are not as clear£ut, but the trend in H1
reflects the interaction of these two components of diversity.
There is no statistically significant correlation between distance from
the power plant and H1 , E, or S in the baseline data (Table 6.33). For all
three least squares regressions, the coefficient of correlation is less than
0.7 (p > 0.05). Total across-season diversity ranges from 1.28 at site 5 to
3.92 at site 39. In temperate regions bird species diversity is correlated
with foliage height diversity (MacArthur and MacArthur, 1961). Diversity
increases with the number of layers in vegetation and with the evenness of
foliage apportionment among layers. Site 5 is an open grassland, one-layered
habitat. Site 39 is riparian. Grassland, shrubs and trees occur within 400 m
providing a three-layered terrestrial habitat as well as an aquatic habitat for
shore and water birds. It may be possible to account for much of the site
variation in bird species diversity by variations in habitat diversity. An
attempt is being made to quantity habitat diversity at the study sites during
the 1976 field season.
Average cross season H1 and S for birds in the pine savannah southeast of
Colstrip are intermediate between typical values for grasslands and those for
shrublands (bottom of Table 6.33). This is expected since the sampling sta-
tions range from open grassland to forest.
Average E is lower than would be predicted by the trends for H' and S.
This may be an artifact of the census method which tends to overestimate
meadowlark abundance relative to other species yielding generally lower E
values than would otherwise be found. Variability in H', S, and E is very
similar to that found in other grassland and shrubland censuses (Tramer, 1969).
H' is dependent upon both species richness and equitability. In general,
bird species diversity correlates strongly with species richness while equit-
ability tends to be high with little variability regardless of species rich-
ness. For 267 breeding bird censuses from many habitat types, Tramer (1969)
demonstrated that: H1 = 0.941 (log2S) - 0.251 (r = 0.972). The same trend
occurs in the baseline across season diversity data in the present study. The
least squares regression line (H1 = 0.999 ± 0.075 S.E. x log2S - 0.889 ± 0.255
S.E.) is similar to the general trend demonstrated by Tramer, but H1 and log2S
are not as strongly correlated (r = 0.754).
The components of H1 (S and E) may respond differently on exposure to en-
vironmental stress. In general, high species richness and high equitability
are characteristics of late successional stages (Margalef, 1963, 1968; Odum,
1969). Stressful environments generally permit less diversity to develop than
favorable environments do (Sanders, 1969; Slobodkin and Sanders, 1969).
Environmental stress is primarily determined by (1) the degree of temporal
predictability in environmental conditions and (2) the degree of physiological
26]
-------
TABLE 6.32. SPECIES FREQUENCY ALONG THE CENSUS ROUTE DURING 1975.
Proportional Abundance (x
Common Name 5/6 5/23 6/11 6/30
Mallard 50
Red-Tailed Hawk 17
Golden Eagle 17
Marsh Hawk 66
Prairie Falcon
Peregrine Falcon
Sparrow Hawk 446
Sharp-Tailed Grouse 66
Ring-Necked Pheasant 826
Killdeer 17
Common Snipe 17
Mourning Dove 446
Long-Eared Owl 17
Poor-Will
Common Nighthawk
Chimney Swift
Belted Kingfisher
Red-Shafted Flicker 66
Red-Headed Woodpecker
Eastern Kingbird
Western Kingbird
Cassin's Kingbird
Say's Phoebe 50
Horned Lark
Barn Swallow
Cliff Swallow
Black-Billed Magpie 33
Common Raven
Common Crow 132
Pinon Jay
Black-Capped Chickadee 413
White-Breasted Nuthatch
House Wren
Winter Wren
Brown Thrasher
Robin 33
Wood Thrush
Mountain Bluebird
Cedar Waxwing
Loggerhead Shrike 50
Starling 132
Yel low Warbler
108
27
108
108
755
296
81
243
189
27
243
81
27
135
108
243
54
162
27
27
20
160
20
747 343
27
427 625
242
53 121
107 81
347 81
53
27
187 161
293 444
80
133 60
323
27 40
107 60
240 20
27
53 60
101*) for the given date Seasonal
7/14 7/28 8/11 8/26 9/8 Total
77 27
40
40 51 23 93 301
27
80 26 68 56 164
26 37 27
783 918 1847 935 902
26
102
20
100 26 68 56 301
82
120 204 225 150 55
141 77 90 224 109
20 19
26 109
80 153 113 93
482 332 473
23
40
40 102 93
180 1458 27
26 90 ' 56 273
23
20 23
23 55
20
221 51 37 27
37
20 77
45
20 45
80 51 90 19 27
7
24
5
12
5
2
140
24
339
15
2
787
2
2
10
29
2
92
7
123
131
10
22
2
107
228
17
5
77
249
114
2
5
7
10
196
5
39
5
17
19
56
(continued)
262
-------
TABLE 6.32. (continued )
Common Name
Yellow Throated Warbler
Ovenbird
Yellow-Breasted Chat
American Redstart
Western Meadow! ark
Red-Winged Blackbird
Bullock's Oriole
Brewer's Blackbird
Common Grackle
Brown-Headed Cowbird
Lazuli Bunting
American Goldfinch
Red Crossbill
Rufous-Sided Towhee
Lark Bunting
Savannah Sparrow
Vesper Sparrow
Lark Sparrow
Chipping Sparrow
Clay-Colored Sparrow
White-Crowned Sparrow
N(xlO°)
H'
E
S
Proportional Abundance (x
5/6 5/23 6/11 6/30
17
4645
413
1058
33
331
430
132
33
605
3.14
0.65
28
27
3396
593
350
108
755
189
539
943
108
371
3.66
0.76
28
27
27
3627
640
27
243
53
107
987
160
187
747
27
375
3.59
0.71
33
60
20
3347
444
383
20
262
81
1815
60
343
444
20
496
3.47
0.71
29
101*) for the given date
7/14 7/28 8/11
20
3775
482
1104
20
261
80
20
60
863
40
321
522
60
498
3.40
0.67
33
26
3163
77
2168
102
26
179
510
77
306
995
26
392
3.29
0.67
30
23
3176
90
203
45
90
68
1779
135
405
541
444
3.22
0.67
28
8/26
19
37
2991
467
280
37
75
56
2056
75
393
93
56
535
3.23
0.67
28
Seasonal
9/8 Total
2787
137
27
82
410
2022
219
1667
137
55
368
3.29
0.70
26
10
2
17
5
3448
361
2
605
12
140
22
56
36
36
1165
157
479
465
31
5
5
3.77
0.63
63
263
-------
TABLE 6.33. SPECIES DIVERSITY, EQUITABILITY, AND SPECIES RICHNESS AT THE
SAMPLING STATIONS DURING 1975
Site number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
Distance from
Power Plant
(km)
15.8
15.0
14.2
13.6
12.8
12.0
11.4
10.6
9.9
9.3
8.6
9.0
9. 1
8.5
8.2
7.5
6.7
5.9
1.3
1.9
2.7
3.5
4.5
5. 1
5.8
16.2
15.7
15.2
14.7
14.4
14. 1
14. 1
14.1
14.1
13.9
13.8
13.3
13.0
13.0
13.0
13.0
13.1
13.4
Species
Diversity
(H1)
2.70
1.35
2.47
2.36
1.28
1.56
2.74
2.97
2.89
3.47
3.29
2.83
1.84
1.82
1.61
2.44
2.24
1.69
1.89
1.31
2.20
2.40
2.69
2.03
1.69
1.67
3.38
2.97
2.88
1.82
2.03
3.05
3.29
2.95
2.62
1.64
2.50
3.33
3.92
2.11
1.95
1.63
2.70
Equitabil ity
(E)
.73
.58
.65
.66
.55
.60
.70
.78
.78
.85
.77
.79
.58
.71
.57
.81
.80
.66
.60
.46
.64
.72
.85
.72
.84
.60
.87
.78
.76
.61
.61
.78
.82
.70
.87
.71
.75
.80
.89
.64
.76
.52
.75
Species
Richness
(S)
13
5
14
12
5
6
15
14
14
17
19
12
9
6
7
8
7
6
9
7
11
10
9
7
4
7
15
14
14
8
10
15
16
19
8
5
10
18
21
10
6
9
12
(continued)
264
-------
TABLE 6.33. (continued)
Site number
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Distance from
Power Plant
(km)
13.9
14.2
14.6
14.9
15.2
15.7
15.8
16.2
16.8
17.3
17.4
17.6
17.8
17.9
18.2
18.2
18.2
Species
Diversity
(H1)
3.09
2.70
2.78
2.26
2.32
2.45
2.40
2.47
3.56
2.84
2.91
3.03
2.80
2.48
2.62
2.95
2.22
Equitability
(E)
.89
.78
.78
.65
.67
.77
.69
.69
.94
.86
.81
.84
.78
.72
.76
.80
.64
Species
Richness
(S)
11
11
12
11
11
9
11
12
14
10
12
12
12
11
11
13
11
Average ± 2 Standard Errors 2.47 ± 0.16 0.73 + 0.02 10.95 ± 0.98
Typical Grassland Values1 1.9310.24 0.84 ±0.034 5.74 ±1.00
Typical Shrubland Values1 3.14+0.16 0.85+0.024 14.08 ±2.31
1from Tramer (1969)
265
-------
stress imposed (Slobodkin and Sanders, 1969). Either a temporally
unpredictable or a temporally constant but physiologically stressful
environment should constrain the development of high species richness since
relatively few species would be able to successfully colonize the environment.
However, equitability in the distribution of individuals should be different in
the two environments. In a constant but severe environment, equitability
should be relatively high in late successional stages. As the species which
have successfully adapted to the extreme environment begin to realize full
exploitation of their available resources, competition may become the dominant
selective pressure and this should lead to greater equitability (Emlen, 1973).
In an unpredictable environment, however, catastrophic disturbances may keep
the ecosystem at a low state of maturity and should lead to a log-normal
distribution of species abundances (MacArthur, 1960). As a result, both
species richness and equitability would be low.
Figure 6.11 illustrates the combinations of cumulative S and E values
which occurred at the 60 sampling stations during 1975. The quadrants
represent the trends in species discrimination of their environments at climax
expected from the previous discussion. This format may prove useful in
interpreting shifts in positions of the plotted points that occur in the
future. Vectors drawn from the baseline point positions for a particular site
to its new position at some future time can be used to quantify the reaction of
the avian community at that site to changes in environmental conditions. The
vector sum for all 60 stations can be used to quantify the new shift away from
baseline of the avian community along the census route and may provide a useful
index of initial response to operation of the Colstrip power plants and permit
tracking of subsequent ecological succession.
Table 6.34 illustrates the interpretation that can be given to various
temporal shifts in vector direction. Vector length can be used to quantify the
magnitude of the vector shift. This technique permits evaluation only of
quantitative shifts in species numbers and relative abundances. Qualitative
changes in species composition may be of equal or greater importance.
Comparisons of the life history strategies of those species favored by
pollution with those less favored may lead to better understanding of more
general effects manifested by the ecosystem of interest. Thus where generalist
species tend to be replacing or increasing at the expense of specialists,
retrograde succession may be indicated.
HISTOLOGY
Avian Tissues As Indicators of Pollution Challenge
Histological evaluation of selective tissues and organs of birds is in
progress. Baseline definition will be complete in about a year. Only those
tissues that are likely to respond to the low levels of emissions expected from
the Colstrip plants are examined histologically. These include the following:
Tissues that are indicators of "stress." The responses of mammals to stress
are well documented (e.g., see Christian, Lloyd, and Davis, 1965) and generally
fit the "general adaptation syndrome" originally promulgated by Hans Selye (von
Faber, 1964). Early responses occur in the adrenal gland, particularly in the
266
-------
CD
O
UJ
c
O
t»
O
0)
(0
0)
tA
O
k_
O
O
UJ
CONSTANT OR
CONSTANT-FAVORABLE
S
E
uu
60
40(
— r-rrcvns IMOI-C. O//TCOO
. • 1
. :••!*!*
• *
• *
UNPREDICTABL E*S TRESS
* I
UNEXPECTED
I I I
•) 5 10 !5 20 25
TOTAL NUMBER OF SPECIES
OBSERVED AT SITES DURING 1975
o
Figure 6.11.
Species richness versus cumulative equitability for the 60
sampling stations during the baseline period (1975).
267
-------
TABLE 6.34. INTERPRETATION OF TEMPORAL SHIFTS IN VECTOR DIRECTIONS
ON PLOTS OF EQUITABILITY VS. SPECIES RICHNESS
A A Directional Shift
Species Richness Equitability in Vector Interpretation
(S) (E) (Degrees)
o + 0 numerically dominant species becoming less
dominant
+ + >0, <90 new species are being added, but all species
are becoming more even numerically
+o 90 new species are being added but relative
abundances have not changed
ro
S
new rare species are being added
>90, <180 numerical dominance
i ncreasing
by abundant species
is
180 numerically dominant species are becoming
more dominant
>180, <270 rare species decreasing and/or dropping out,
dominant species may be increasing
no differential effect on relative abundance
270 all species decreasing numerically, rare
species dropping out
(1) rare species drop out, others remain
>270,<360 abundant
(2) rare species drop out, dominant species
become less abundant
-------
adrenal cortex; this tissue hypertrophies and produces unusually large amounts
of glucocorticoids. Lymphatic organs involute secondarily and predictable
changes also take place in the blood (including lymphocytopenia and
neutrophilia). These functional changes are accompanied by changes in tissue
structure.
1. Adrenal Gland—There are surprisingly few studies of stress-related
changes in the adrenal glands of birds, but those available are in harmony with
the above picture of stress on the mammalian adrenal gland. Stressors such as
oil, kepone, and high population density are known to immediately increase the
weight of the gland (Flickinger, 1961; Gorman and Milne, 1971; Eroschenko,
1973). However, the adrenal gland of the bird is a mixture of interrenal and
chromaffin tissue and changes in weight indicate little about which component
is affected. In fact, changes in the weight of the gland are sometimes
misleading. Seasonal changes in the adrenal gland of the Wood Pigeon provide
an instructive example (Ljunggren, 1969). To begin with, there are no marked
seasonal changes in the weight of the gland. Yet, on a histological level,
there are rather pronounced seasonal variations in the activity of interrenal
and chromaffin tissue. Furthermore, the male adrenal is generally heavier than
that of the female. On a weight basis, it might be concluded that males are
stressed more than females. However, just the opposite appears to be true:
the fractional volume of cortical tissue in the female's gland always exceeds
that in the male's gland.
Among the few histological studies that exist concerning birds, stress-
related changes in the adrenal include (1) hypertrophy and hyperplasia of in-
terrenal and chromaffin cells, (2) changes in the fractional volumes of inter-
renal and chromaffin tissue, (3) increases in the number and size of nuclei
within interrenal cells, and (4) increases in the vascularity of the gland
(Stoewsand and Scott, 1964; Ljunggren, 1969; Gorman and Milne, 1971;
Bhattacharyya and Ghosh, 1972; Bhattacharyya, Ray, and Manna, 1972;
Eroschenko, 1973). The subject is well reviewed by von Faber (1964).
Few seasonal studies of the adrenal gland exist and the functional signi-
ficance of seasonal changes in this gland are not well established. In all
species examined to date except the Brant, adrenal activity is reduced during
periods of molt, but elevated at breeding (summarized in Gorman and Milne,
1971). Changes in the adrenal weight of Western Meadowlarks and Mourning Doves
during 1975 at Colstrip (Lewis, unpublished) are consistent with these earlier
studies— adrenal weight increased during the breeding season and diminished
during the postnuptial molt.
2. Thyroid Gland—Manifestations of stress at the level of thyroid gland
include losses of colloid and increases in the height of the follicular
epithelium. Both changes are indicative of elevated glandular activity.
Little, if any, concurrent change takes place in the weight of the gland
(Ljunggren, 1968) which underscores the need for its histological evaluation.
The adrenal and thyroid glands at the tissue level are potentially sensi-
tive and quick-responding bioindicators of (1) the normal stressors in the
Colstrip ecosystem and (2) the additional stress of power plant emissions.
Blood levels of corticosterone in key avian species will be measured in future
269
-------
summers at the site. It will be interesting to compare changes in the histol-
ogy of the interrenal tissue and alterations in corticosterone production.
Tissues that belong to the immune system. Tissues in this category react
directly to foreign materials in the body or are secondarily affected by the
induced activity of the adrenal gland.
1. General—The immune system of the bird includes lymphoid cells that
originate in the thymus (T lymphocytes), bone marrow (BM lymphocytes), and
bursa of Fabricius (B lymphocytes). These cells disseminate to lymphatic tis-
sue throughout the body and are responsible for immune responses. Descendants
of T cells produce substances which destroy foreign bodies, such as skin
grafts, outright. Descendants of BM and B cells produce nonspecific and
specific antibodies, respectively. There is accumulating evidence that T cells
are also involved in activities traditionally ascribed to BM and B cells. The
entire subject of avian immunity was reviewed recently by Moticka (1975).
2. Bursa of Fabricius—The bursa is a lymphoepithelial gland which deve-
lops as an outgrowth of the proctodaeum. It is necessary for the development
of antibody-mediated responses. It normally involutes when a bird reaches
sexual maturity. However, it will involute earlier if the younger bird is
subjected to stress (von Faber, 1964). It will, for example, regress in the
presence of glucocorticoids (Click, 1964; Siegel , 1971). Consequently, it
should be very useful for identifying stressors that affect young birds, as
opposed to those which affect the avian population as a whole. Sudden
regressive changes in the structure of the bursa are more likely to be
translated quickly into histological changes than into reductions in the weight
of the gland. This underlines the importance of examining the tissue structure
of the gland.
3. Thymus—The thymus is a multilobed gland which is distributed along
the jugular veins in the neck. It is diffusely arranged and difficult to re-
move totally, though we are generally able to do this successfully. The thymus
of many avian species, including the Mourning Doves and Lark Buntings collected
at Colstrip (Lewis, unpublished), enlarges during molt (Hb'hn, 1956; Anderson,
1970; Kendall, 1975; Ward and Kendall, 1975). Periodic rejuvenation of the
gland at this time appears to provide erthrocytes to meet the demands of molt
and perhaps additional lymphocytes to deal with stresses additional to that of
the molt itself (Ward and D'Cruz, 1968; Ward and Kendall, 1975). T«he thymus
involutes if the adrenal gland becomes hyperactive. Immediate and predictable
thymic responses include lymphopenia, eosinopenia, and heterophil ia.
Consequently, it may be particularly sensitive to the stress of chronic, low-
level pollution challenge. Stress-related effects are more likely to be
registered first as alterations in tissue structure and only later as changes
in organ weight.
4. Spleen—The spleen has been largely ignored by avian biologists (Ewart
and McMillan, 1970). Since it is not a storage organ for blood, changes in its
weight may be true reflections of different states of its activity. In this
respect, some literature concerning seasonal changes in its weight is available
for reference (Riddle, 1928; Oakeson, 1953). Its major functions are
haemopoiesis, antibody production, and filtration of the blood (as part of the
270
-------
mononuclear phagocyte system). Splenic leucopoiesis is influenced by the
activity of the adrenal gland and by antigen challenges. Foreign proteins and
carbohydrates, for example, stimulate its production of plasma cells and
antibodies (Jankovic and Isakovic, 1966). The phagocytes of its red pulp also
selectively engulf blood-borne particulate matter. The number and activity of
the germinal centers and the activity of the follicular tissue in its white pump
are directly related to its antibody production. In addition, the activity of
parafollicular areas of its white pulp varies directly with the number of cell-
mediated immune responses taking place within the birds. In other words,
splenic tissue provides information about the magnitude of immune responses and
also permits one to further determine if the responses are primarily humoral,
cell-mediated, or both.
The value of the spleen as an early indicator of pollution challenge is
suggested by the studies of Zarkower (1972) with mice. He demonstrated that air
containing carbon, sulfur dioxide, or combinations of the two, suppresses immune
responses. These pollutants destroy cells of the immune system selectively
(Piliero, 1970) and their effects should be expressed relatively early in the
structure of lymphoid tissues.
Detoxifying tissues. The major site of detoxification in the vertebrate
body are the liver, kidney, and lungs.
1. Liver—The liver has a large number of functions, only one of which is
the detoxification of endogenous and foreign substances which pass through it.
These other functions probably reduce its utility as a specific indicator of
pollution challenge. This is illustrated by the fact that liver weight varies
with age, diet, season, stage of the reproductive cycle, and even time of the
day (Farner, 1960; Hartman and Browne!1, 1961; Ljunggren, 1968). Weight
variations such as these underscore the importance of examining hepatic tissue
as a possible indicator of pollution effects, rather than relying on changes in
its weight which are relatively insensitive at best.
Changes in hepatic structure of birds in the Col strip area in response to
chronic exposure to trace metals and other pollutants may occur. Toxicants such
,as ethanol and carbon tetrachloride produce cirrhosis; and kepone causes focal
necrosis and marked congestion in the liver (Huber, 1965). Trace metals, such
as arsenic and copper, that occur in stack emissions, produce fatty degeneration
and necrosis of hepatic parenchyma (Smith and Jones, 1961). Gases such as CO,
also found in stack emissions, directly suppress the detoxifying functions of
rat liver (Cooper et al. , 1965; Conney et aj_. , 1968), whereas chlorinated
hydrocarbons such as DDT enhance the catabolic functions of avian liver (Peakall
1970). In addition to direct pollution-related alterations of hepatic
parenchyma, the liver's population of Kupffer cells (as part of the mononuclear
phagocyte system) may accumulate foreign particulate material at rates that
correspond to environmental pollution levels.
In summary, the possibility that the liver will exhibit histological
changes in birds from chronically impacted areas is very real and needs exam-
ination. In addition, the liver may be useful for distinguishing between the
effects of ingested pollutants and those acquired via the respiratory system.
271
-------
2. Kidney—The kidney is the second major site of detoxification in the
vertebrate organism as well as the major site of excretion. It is directly
damaged by pollutants such as copper, fluorine, lead, and mercury, all of which
occur in stack emissions (Smith and Jones, 1961). Damage to the kidney's
filtration apparatus is indicated by the presence of protein (casts) within
renal tubules. There may be complete loss of renal tubules, or simply
disintegration of their epithelia. Congestion, the accumulation of fluid
between tubules, and the presence of fibrin in the renal corpuscule are other
common pathologies. Reliable and thorough histological descriptions of the
avian kidney are now available for reference (Johnson and Mugaas, 1970 a,b).
3. Lung—The respiratory tract is a third major site of detoxification in
the vertebrate organism. In addition, its alveolar epithelium contains
phagocytes which remove particulate matter from inhaled air. The lung is sub-
ject to direct irritation from inhaled substances; and because of its unusually
large blood supply, it reacts to these substances immediately with tissue
proliferation, pneumonia, focal accumulations of leucocytes, congestion, and
visible damage to the alveolar wall (Smith and Jones, 1961). The lung's
unusual sensitivity to air-borne pollutants is suggested by Lewis, Morton, and
Jones (1975). The histology of the tissue should consequently be a
particularly sensitive and selective bioindicator of air quality.
Reproductive tissues. The reproductive system is one of the first to be
affected deleteriously by chronic stress (Christian and Davis, 1964; von Faber,
1964; van Tienhoven, 1968). Because this system, particularly the histological
events of spermatogenesis, has been well studied in a large number of birds
(Lofts and Murton, 1973), any deviations from normal in the Col strip population
should be readily appreciated.
The importance of evaluating reproductive tissues is twofold. In the
short term, they should be relatively sensitive, although secondary,
indicators of the stress associated with pollution impact on the ecosystem. In
the long-term ecological context, they are equally important predictors of
population growth in chronically polluted areas, since upsets in the
reproductive function of one generation directly influence the size of the
following generation.
ACKNOWLEDGEMENTS
The authors wish to thank the several people who have served as technical
assistants over the past two years. A special thanks is extended to Larry Doe
and Tom Gullett for their major assistance during field operations.
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279
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SECTION 7
REMOTE SENSING OF THE BIOENVIRONMENTAL
EFFECTS OF STACK EMISSIONS IN THE
COLSTRIP VICINITY
by
J. E. Taylor and W. C. Leininger
INTRODUCTION
Procedures for ground and aerial photography have been developed to
monitor air pollution effects in a grassland ecosystem. Photography offers
several strong advantages for biological monitoring. Permanent, complete,
periodic records can be obtained, stored, exactly reproduced, and compared over
time. In this way trends and changes can be recognized and documented. Also,
the relatively unselective nature of photography permits re-examination of
past imagery for previously unnoticed features which later studies show to be
important.
The photographic studies described here include groundlevel and
relatively low elevation aerial imagery. Periodic flights allow aerial
assessment of rates of vegetation changes. Ground studies help discriminate
between normal and stress-induced changes. Pollution stress is detectable, but
not yet quantifiable, in low-level aerial photography.
GROUND LEVEL PHOTOGRAPHY
Ground level photography provides a detailed record of plant species,
phenology, and pathologic signs. All of this contributes to the total data set
and assists in the interpretation of aerial imagery. Also, vertical ground
photo plots may be measured and analyzed for cover, number, frequency, pattern,
and plant volume.
PROCEDURES
Photo plots are established on all study sites and on representative
examples of the various communities present in the area. Permanent meter-
square plots are photographed stereoscopically in color and black-and-white.
Details are given by Taylor et aj_. , 1976.
Aspect photographs are made from vantage points within and overlooking
plot areas. These are taken with color and infra-red color, the latter to
compare with aerial coverage.
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Ground photography is collected periodically through the growing season
so that any changes will be recorded.
DISCUSSION
A sample stereoscopic pair of photo plots appears as Figure 7.1. Species
identification, canopy cover, litter, bare ground, and plant heights are among
the kinds of data which can be derived from these photographs.
Plant volume, which can be used to estimate biomass, may be obtained by
combining canopy coverage and height, the latter measured with a parallax
wedge. Plant density and pattern also can be studied from these pictures.
For each photo plot an index of species identification has been prepared.
An example is shown as Figure 7.2. Species which are diffused throughout the
quadrat are termed "matrix species," and are listed but not precisely located
on the indices. Species which are readily distinguishable in the plot-
photographs are mapped. The combination of plot photographs and plot indices
makes a permanent record of species presence and distribution. Sequential
records allow the evaluation of temporal changes.
AERIAL PHOTOGRAPHY
Low level aerial photography gives a more generalized view of plant
species and community distributions, pathology, and cover than does ground
photography. However, it yields more detail than higher level imagery and so
represents a useful compromise between detail and comprehensiveness of
coverage.
Low level aerial imagery is most practical for making detailed vegetation
maps, sensing population-level stresses, or any other purposes requiring large
scale synoptic views. It also aids in developing interpretations of smaller
scale, high level photography.
PROCEDURES
In this project, a Cessna 182 airplane which can easily handle the
elevation range of 500 to 7000 feet above ground, was used. The plane, leased
from Miles City Flying Service, Miles City, Montana, has been modified by the
addition of a 12" diameter belly hole, which accepts a special mount, designed
and manufactured by W.E. Woodcock of Miles City (Woodcock, 1976). The mount
supports a Hasselblad EL/M motor-driven camera (70 mm format) and a manually-
operated Leica MDa (35 mm). Both are fitted with 50 mm lenses, yielding
negative scales from 1:3000 to 1:40,000. Other focal lengths are sometimes
used to obtain changed scales without changing flight elevation. For special
purposes, 80, 150, 250, and 500 mm lenses are used.
In addition to the mounted camera, oblique photographs from the air are
taken with a second Hasselblad used as a hand camera. This kind of photography
supplements the more traditional vertical imagery, since it is more
representative of familiar aspects of scenes for interpretation, display, etc.
281
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ro
8
Figure 7.1. Example stereogram of photoplot (McRae Knoll B, June 16, 1975).
-------
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OJ
0 / 2 3 4 5 6 7 8 9 IO
1
2
3
4
S
6
7
8
9
in
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/ 1
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- \ /
~^~ — — ' *z
SYMBOL
—
Z
C
F
K
S
B
A
Agropyron spicatum
Allium textile
Artemisia cana
Artemisia frigida
Car ex fill folia*
Ceratoides lanata
Koeleria cristata*
Leucocrinum montanum
Sphaeralcea coccinea
Stipa comata*
Taraxacum officinale
Tragopogon dubius
(*MATRIX SPECIES)
Figure 7.2. Photoplot index of McRae Knoll B, June 17, 1975.
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DISCUSSION
Three primary film types are used with other materials for special
purposes. The main films are color, color infra-red and black-and-white (H&W
VTE Pan). Each has advantages for particular uses.
Color is realistic and many plant species are easily recognized. It is
easy and fast to process, and is visually acceptable to lay viewers.
Color infra-red is excellent for delineating plant species and stress
effects. Although it appears strange to inexperienced users, it is useful for
many aspects of this project.
Black-and-white is most practical for mapping photography, especially at
relatively small scales. It makes a good, inexpensive base map for overlaying
ground information (vegetation, geologic features, soil types, drainages,
etc.).
The Hasselblad format (6x6 cm) allows greater coverage than the 35 mm
(2.4x3.6 cm) frame. Especially at low elevations (<1000 ft), this increases
the chance of covering ground targets completely. The 35 mm photography
permits processing in the field, so that pictures taken one day can be
interpreted the next. Also, if exposures or coverage are not acceptable, a re-
flight can be made immediately. When the larger format photography is returned
from processing, the interpretations on the 35 mm imagery can be applied.
An example of low level aerial photography is given in Figure 7.3.
General plant pattern, landforms, deferment effects, and sampling impacts are
apparent. This kind of photography can be analyzed for plant cover, pattern,
and identification of conspicuous species.
COLSTRIP
Investigators working on this aspect of the Colstrip project have gained
enough experience to make detailed vegetation maps from aerial photographs with
only limited ground checking. Not only can they separate gross vegetational
groupings (trees, shrubs, grasslands), but they are also able to discriminate
among grassland types at a high degree of resolution.
Figures 7.4, 7.5, and 7.6 are vegetation maps of the McRae Knolls. Thirty-
six mapping variables are indicated; some are plant community types and others
are individual species. Table 7.1 is an index to map units.
The base for this map was 1:660 color aerial photography. A ground
transect network provided criteria for mapping units and identification. This
map constitutes a baseline for mapping other sites in the general area.
The investigators conclude that four overflights (early spring, peak of
green for cool-season species, peak of green for warm-season species, and
summer dormancy) using both color and color infra-red film types provide an
adquate base for detailed vegetational mapping in heterogenous situations.
284
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ro
8
Figure 1.6. LOW levei aeriai pnotograph or Hay Coulee exclosure.
-------
CD
CT)
Figure 7.4. Detailed vegetation map, McRae Knoll A, Summer, 1976.
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INi
Figure 7.5. Detailed vegetation map, McRae Knoll B, Summer, 1976.
-------
ro
CO
CO
Figure 7.6. Detailed vegetation map, McRae Knoll C, Summer, 1976.
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TABLE 7.1. INDEX TO MAP UNITS.
SYMBOL
1
2
3
4
5
6
7
8
9
10
n
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
36
27
28
29
30
31
32
33
34
35
COMMUNITY
Tree types
Acer negunda
Fraxinus pennsylvanica
F. pennsyl vanica/Prunus virginiana
Juniperus scopulorum
Populus deltoides
Primus americana
P. virginiana
Prunus/Acer/Fraxi nus
Salix amygdaloides
Shrub types
Artemisia cana
A. cana/mixed grass
Artemisia dracunulus
Ceratoides lanata
Rhus trilobata
R. trilobata/P. virginiana
R. trilobata/Ribes/P. virginiana
Ribes
Rosa woods ii
Shepherdia canadensis
Symphoricarpos occidental is
S. occidental is/A, cana
S. occidental is/Ribes
S. occidentalis/Ribes/R. woodsii
S. occidental is/R. woodsii
S. occidentalis/R. woodsii/Ribes/P. virginiana
Forb types
Yucca glauca
Mel i lotus officinal is
Grass types
Bromus inermis
B. japonicus/B. tectorum
Carex filifolia/B. japonicus
Calamovilfa longi folia
C. longifolia/C. filifolia
Poa pratensis/Agropyron smithii/Stipi viridula
P. pratensis/Carex
P. pratensis/S. viridula/A. cana/A. smithii
Schizachyrium scoparium
289
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In more homogenous vegetation, additional ground studies are needed to
elucidate the more subtle community patterns. The Kluver sites and Hay Coulee
fall in this category. Larger scale photography and further ground
observations are planned for these locations.
ZAPS
Even though the aerial photography shows distinct tonal differences among
the ZAPS plots, the reasons are still to be determined. Early signs of S(L
stress can be detected, but the magnitude of pollution damage cannot be
quantified. Additional ground studies and more detailed photographic analysis
should lead to an accurate and fast method for damage assessment.
SUMMARY
Photo plots have been established throughout the study area, and are
examined periodically during the growing season. They provide records or
changing species composition, cover, and stress effects.
Aerial photography techniques have been developed to monitor air
pollution effects as shown by plant changes. Color, color infra-red and black-
and-white films each have particular uses.
Pollution stress is detectable in aerial photography, but cannot be
quantified without additional analysis.
A procedure for vegetational mapping has been developed and tested.
REFERENCES
Taylor, J. E. , W. C. Leininger, and R. J. Fuchs. 1976. Monitoring plant
community changes due to emissions from fossil fuel power plants in
eastern Montana. Section II of the bioenvironmental impact of a coal-
fired power plant, second interim report. USEPA Ecological Research
Series EPA-600/3-76-013. pp. 14-40.
Woodcock, W. E. 1976. Aerial reconnaissance and photogrammetric with small
cameras. Photogrammetric Eng. and Remote Sensing.42:503-511.
290
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SECTION 8
INTEGRATED AEROSOL CHARACTERIZATION MONITORING,
COLSTRIP, MONTANA
by
N. L. Abshire, V. E. Derr, G. T. McNice, R. Pueschel, and C. Van Valin
INTRODUCTION
GENERAL
In addition to other objectives, the Colstrip Coal-Fired Power Plant
Project requires the concentration, dispersion and identification of background
particulate matter and particulate matter resulting from the activities of the
mines and coal-fired power plants. This project started in May 1975.
Observations were performed at Hay Coulee from May 18 to June 16, 1975; from
August 17 to September 15, 1975; and from May 23 to June 5, 1976. In addition,
aircraft flights were made on April 20 and 21, 1976. The observation periods
were chosen to give maximum density of observation in the Spring and Fall at the
beginning and end of the growing season. Beginning in October 1976 a sampling
flight was made between the intense observation periods. Further intense
observations were made August 14 to August 28, 1976, and in the Spring and Fall
of 1977.
This research program is designed to help the Environmental Protection
Agency Laboratory at Corvallis, Oregon, assess the impact of coal-fired power
plants on a grassland ecosystem and develop a valid and effective environmental
impact assessment protocol. Atmosphere particles emitted by coal-fired plants
(e.g., ash) or formed from gases emitted (e.g., sulfates or chlorides) produce
an array of effects on the ecosystem. These particulates may produce direct
biolgical effects by contact through leaves and soil. They may also affect the
radiation reaching the biosphere by absorption and blanketing mechanisms.
Further, by adding cloud condensation nuclei and ice nuclei to the atmosphere,
they may significantly modify precipitation, either near the source or far
downwind. The effects of particulate matters are strongly dependent on the
temperature structure (inversions) and mixing and diffusion characteristics of
the atmosphere downwind from the source. The parameters needed to characterize
the effects of particles should be examined over a wide geographical area and
over sufficient time to obtain representative averages.
The program seeks to characterize the impact of air pollutants on the total
ecosystem. Small particles are a significant part of the pollution emitted by
fossil fuel power plants, even when the stack emission has been carefully fil-
291
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tered. These particles from the stacks and the natural background of particles
from the earth's surface are called "aerosols." For this report, clouds and
fogs are excluded from the term aerosols. Aerosols range in size from a clump of
a few molecules to tens of micrometers. The larger aerosol particles fall out
rapidly and are not as frequently observed in the atmosphere. However,
particles up to several micrometers in diameter may remain suspended in the
atmosphere for long periods, up to weeks or months. Thus if filters which
remove large particles are not as effective in removing small particles, the
plume from a smoke stack may produce significant pollution which can remain
suspended in the vicinity, or be swept by winds over distances up to several
hundred miles. In order to evaluate the amount of such material, its source,
its distribution in the atmosphere, the effects of local weather conditions and
terrain, and to identify the composition and size and shape distributions of the
particles, it is necessary to use special remote, i_n situ and airborne sensors.
These sensors are described in some detail in the Discussion section of this
paper. The principles of such measurements are discussed below.
MEASUREMENT RATIONALE
Particles
The remote measurement of the distribution of particles in the atmosphere
is accomplished by the use of lidar (Lujht Detection and Ranging). Lidar
operates very much like radar. A powerful short pulse of light (10"8sec) is
transmitted. The light scattered back from small particles is collected by a
telescope (70 cm diameter) and detected by photomultipliers. Lasers, operating
at near visible wavelengths are required to detect particles whose radii are of
approximately the same size as the electromagnetic radiation (light) waves
scattered. If the particles are small compared with the wavelength of the
radiation (as would be the case for microwave radar) the scattered radiation is
too weak to detect. Detection of particles by a calibrated lidar yields two
pieces of information. First, the backscatter cross section is obtained. It is
dependent on the size, shape and index of refraction of the particles and is
proportional to the number of particles of a given size, shape and index. The
chief asset of lidar remote sensors is the ability to obtain signals from
particles at large distances, up to 20 km, and by scanning the vertical
hemisphere, with good spatial resolution, to determine the distribution of
particles. Second, the polarization of the scattered light is measured. Since
the laser transmitter is linearly polarized, the degree of depolarization caused
by the scatterer may be determined. •
The degree of depolarization is indicative of the shape of the particle:
spherical particles produce no depolarization in backscatter; non-spherical
particles can cause significant depolarization. Since newly-disturbed dust
particles in the atmosphere are frequently very non-spherical, the depolari-
zation is frequently a useful way of discriminating between dust from the mines
and the more spherical particles from the stacks.
The lidar cannot measure mass or particle concentrations without cali-
bration. For that purpose, aircraft sampling is employed. Suitable nucleopore
or millipore membrane filters capture particles for later elemental, size, and
shape analysis with an x-ray fluoresence spectrometer and an electron
292
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microscope. The aircraft also contains an Aitken particle counter which
determines the number of particles per liter With these calibrations, and
since the variation of aerosol types in time and space is small compared with
aircraft sample times, the lidar may be used efficiently to measure mass
concentrations over very large volumes and observe changes in those
concentrations under the influence of local weather conditions.
The very important identification of particle characteristics is performed
by several instruments located on the ground in a trailer next to the lidar
system. This system, of fundamental importance to the project, measures many
parameters of the particles. The trailer is equipped to continuously measure
the total aerosol population, or "Aitken nuclei," (active at 300 percent
supersaturation), cloud condensation nuclei (active at 0.1 percent
supersaturation), aerosol light scattering and standard meteorological para-
meters such as wind speed, direction, temperature, and humidity.
The direct measurement of solar radiation at visible wavelengths can
provide an independent determination of aerosol loading. Radiation from the sun
in the visible range is known to remain constant to within 1 percent over
periods of decades. Observation of solar irradiation by instruments at ground
level can therefore be used to accurately measure the attenuation imposed by the
atmosphere. The major causes of atmospheric attenuation (turbidity) are clouds,
aerosols, ozone, water vapor, and Rayleigh scattering by the air. The effect of
Rayleigh scattering can be accurately calculated and allowed for Ozone and
water vapor absorption occurs in specific wavelength bands and can thus be
avoided or corrected by proper choice of wavelength band for the observing
equipment. Therefore, when clouds are not between the sun and observer, the
attenuation by atmospheric aerosols alone can be determined.
Radiation Effects
Two radiation measurements are of importance. First, the rate at which
solar energy reaches the earth as a function of wavelength must be measured
while the power plants are not in operation (baseline measurements). The
changes registered during succeeding measurements while the plants are
operating may then be correlated with the particulate loading and changes in
cloud cover. Second, the net heat loss of the earth must be measured by infrared
radiometry by measuring the upward and downward radiation, employing aircraft
and ground-based instruments, operating from 8 to 12 pm.
Measuring the IR fluxes within and outside the plumes gives the IR
"shadowing" effect by the plume, inversion techniques permit the determination
of the IR volume absorption coefficient. In conjunction with the measured
aerosol size distribution, the aerosol absorption index can be evaluated, aiding
in the evaluation of long-term climatic effects.
Meteorological Conditions
Standard meteorological variables are measured by standard instruments.
The meteorological conditions which trap pollutants below temperature
inversions are best measured by an acoustic sounder. The acoustic sounder is
also a radar-like device, which uses a pulse of sound. The scattering of sound
back to the receiver is dependent on the degree of homogeneity in vertical
293
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temperature structure. The device is especially sensitive then to temperature
inversions and can determine their height and persistence.
ENVIRONMENTAL EFFECTS
Environmental and health effects of aerosols produced by human activities
and by industrial activities in rural Montana are foreseen in four areas:
1. Inadvertent weather modification: Size, shape and water solubility
(chemical composition) determine the cloud and ice nucleating capabilities, and
thereby effects of the atmospheric aerosol on cloud and precipitation processes.
The proposed experiments are designed to scrutinize a cause-effect relationship
between weather modifying particular matter and human activities in Montana.
2. Climate modification: Size, shape and chemical composition
(refractive index) determine the light scattering and absorption capability of
the atmospheric aerosol. The backscattering to absorption ratio, in relation to
the surface albedo, is the critical parameter that determines whether the
aerosol's effect leads to cooling or warming of the earth-atmosphere system. IR
extinction can cause surface temperature changes by thermal blanketing and
affect radiometric measurements from satellites. All of these parameters will
be evaluated by means of the experimental data.
3. Ecological effects: The damage to vegetation by sulfur and halogen
gases and to structures by corrosive particulate sulfates and halides downwind
from their sources is greatly affected by the gas-to-particle conversion rate.
Ir\ situ measurements of this conversion rate as function of the amount of water
vapor and third-body aerosols is planned by adding proper gas analysis
techniques to the measurements proposed.
4. Health effects: Particle sizes determine the depth of penetration of
an aerosol into the animal respiratory system. Particle solubility (chemical
composition) determines their retention times. The data from the proposed
research will enable health officials to better evaluate health hazards
resulting from environmental pollution.
GENERAL DISCUSSION OF OBSERVATIONS
Observations were planned for Spring and late Summer of 1975 to provide a
baseline against which to judge changes in atmospheric aerosols aft*r the coal-
burning plants began operation. Further observation was planned for Spring and
late Summer 1976 and 1977. The observation of May 23 to June 15, 1976, provided
opportunity only for measurements of unit #2 operating under initial tests with
gas rather than coal. Interesting differences from 1975 observations were
observed which can only be partially reported here until the data processing is
complete.
294
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MEASUREMENT TECHNIQUES
GENERAL APPROACH
In order to adequately characterize the aerosol content, a combination
of observation techniques was used. The method was to make well calibrated
in situ and airborne measurements to provide benchmarks for some of the
remote sensing devices, primarily the lidar (laser radar), enabling them to
extend the calibrations throughout the volume of interest. The various
devices and techniques are described below and are summarized in Tables 8.1
and 8.2.
SURFACE MEASUREMENTS
1. Aerosol collection: Aerosols were collected on membrane filters
and later examined for number, shape and size distribution in a transmission
electron microscope, for elemental composition (atomic number > 11) in a
scanning electron microscope with energy dispersive x-ray analysis, and for
ice nuclei in a thermal diffusion chamber.
2. Aitken Nuclei (AN) count: AN are defined as the total aerosol
population, and their concentration was measured with a continuously operating
Environment-One particle counter. This unit subjects a humidified air
sample to rapid adiabatic expansion that results in a water saturation of
300 to 400 percent. In this condition, water droplets form on all aerosol
particles present in the chamber. The particle concentration is
proportional to the attenuation of a light beam propagated through the
cloud.
3. Cloud Condensation Nuclei (CCN): Many of the AN are effective as
CCN. Their concentration was measured in a thermal diffusion chamber by the
photographic technique of Allee (1974) at .5 percent supersaturation, a
valve typically found in nature. The cloud droplets found were photographed
and counted.
4. Ice Nuclei (IN): IN comprise a small but important subset of the
AN. These were counted with an NCAR acoustic ice nucleus counter (Langer
et al_. , 1967) at -20°C (Bigg e_t al_. , 1963).
5. Gas analysis: Sulfur dioxide was monitored with a Meloy Sulfur
Gas Analyzer. A Theta Sensors, Inc. sulfur dioxide analyzer was used for
the 1976 airborne measurements.
6. Meteorology: Wind speed and direction, temperature, relative
humidity and insolation were measured continuously.
7. Insolation: The total radiant energy density, including both
direct and scattered sunlight, was continously monitored with an Eppley
pyranometer.
8. Light scattering: The coefficient of scattering of atmospheric
aerosols was measured continuously with the Meteorology Research, Inc. model
1550 nephelometer.
295
-------
TABLE 8.1. PARAMETERS MEASURED IN THE NOAA EPA COLSTRIP INTEGRATED ATMOSPHERIC
CHARACTERIZATION PROGRAM.
ro
to
01
Parameter
Temperature
Wind, Direction, Vel.
Cloud Cover
Relative Humidity
Solar Energy, Direct
Solar Energy, Direct
Spectral
Solar Energy, Total
Turbidity
Lidar Aerosol Scatter
Radar
Atmospheric
Temperature
Turbulence
Radiometer
Photography
Units
"C
M/S, Degrees
%
%
Cal/cm /min
Cal/cnWmin
Cal/cm /min
Volts
Backscatter
Intensity
Backscatter
Intensity
(Uncalibrated)
Backscatter
Intensity
(Uncalibrated)
watts/cm'
Method
of
Recording
Recorder
Recorder
Log
Recorder
Log
Log, Digital
Tape, Photos
Photos
Facsimile
Chart, Tape
Film
Instrument
Pyrheliometer
(no filters)
Pyrheliometer
(w/3 filters)
Pyranometer
(no filters)
Volz
Turbidity meter
Lidar System
Radar System
Acoustic
Sounder
Barnes PRT-5
Camera
Frequency
of
Recording
60, (00)
Continuous
1. 2 times
daily or when
changes occur
2. Whenever
insolation meas.
made
Continuous
Continuous
during day
1000,1300,
1600
Continuous
during day
1000,1300,
1600
In accordance
with variability
Clouds only
Continuous
Chart-contin-
uous tape-on
each shot
As needed
Precision
0.5 C
10%
Accuracy
0.5 C
10%
See FMH - 1
10%
5%
5%
5%
Unknown
±10%
—
—
10%
--
10%
5%
5%
5%
Unknown
±30%
—
—
10%
-
Remarks
....
Wide Band
3 wavelength bands, "when
sun's disk is visible
APCL+WPL
....
Polarization Ratio Available
Test for Cloud Observation
Mark time on Facsimile
Chart
Mark time on Chart
Targets of Opportunity
-------
TABLE 8.2. PARAMETERS MEASURED IN THE NOAA EPA COLSTRIP INTEGRATED ATMOSPHERIC
CHARACTERIZATION PROGRAM.(continued)
Parameter
Cloud Condensation
Nuclei
Cloud Condensation
Nuclei
Ice Nuclei
Ice Nuclei
Light Scattering
Aitken Nuclei
Aitken Nuclei
Cloud Cover
(Insolation)
Sulfur Gases
Elemental Composition
Of Individual Particles
Aerosol Size
Distribution
Atmospheric
Transmissivity
(IR Extinction Coefficient)
Units
CM'3
CM'3
CM'3
CIVT3
bscat<1°-4m-1>
CM'3
CM'3
CM'3
ppb-ppm
Method
of
Recording
Recorder
Photographic
R
L
R
R
I
R
R
PandL
Pand L
R
Instrument
MRI
Allee
Photographic
NCAR Acoustic
Counter
Langer Diffusion
Chamber
MRI
Env. -1
Gardner Counter
Eppley
Pyranometer
Meloy
Electron
Microscope
(SEM-EDX)
Electron
Microscope
(SEM-EDX)
Barnes
Radiometer
Frequency
of
Recording
Continuous
Point
Sample
C
PTI
C
C
Point Sample
C
Preselected
Time
Intervals
Preselected
Time
Intervals
Airborne
Measurements
Precision
±20%
±10%
±50%
± 5%
±10%
±10%
•-
±0.01 ppm
For Heavy Metals
Approximate
Sensitivity 10"15gm
±10%
0.5 C
Accuracy
Unknown
Unknown
but high
Unknown
(State of the Art)
Unknown
(State of the Art)
± 5%
±10%
....
....
±0.01 ppm
±10%
0.5 C
Remarks
Visual Counting-
Absolute Measurement
Including Airborne
Collection by
Membrane Filter
—
Ground and Airborne
Measurements
—
Including Airborne
Measurements
Membrane Filter Collection--
Ground and Airborne
Membrane Filter Collection-
Ground and Airborne
Downward Looking
-------
AIRBORNE MEASUREMENTS
A Cessna 182 single engine aircraft was utilized for the measurement of
temperature, AN concentration (using a Gardner Small Particle Counter) and
the collection of aerosols on membrane filters as described earlier. A
Barnes PRT-5 radiometer was utilized for the measurement of atmospheric
transmissivity looking down.
REMOTE SENSING MEASUREMENTS
1. Lidar: The remote measurement of atmospheric particle density was
accomplished through the use of Lidar. A short powerful pulse of light is
transmitted into the atmosphere. Some of this light is scattered by the
atmosphere back into a telescope, detected and processed, yielding the
optical backscatter coefficient of the scatterer. This quantity is propor-
tional to the number of scatterers of given size, shape and refractive
index, but cannot produce either mass concentration, size distribution or
chemical composition without the support of the i_n sj'tu and airborne measure-
ments. For a given situation, however, it is very sensitive to any changes,
particularly in concentration, and has proved very useful in alerting the
other observers as to a change in the character of the atmospheric aerosols.
The lidar system consists of a semi-trailer and a van (Figure 8.1). The
lasers and telescope arejocated in the semi-trailer. The laser generates a
5 joule pulse, at 6943 Angstroms wavelength, 30 nanoseconds in duration at
repetition rates up to 1 pulse per second. The beam is collimated to 1
milliradian beamwidth and aligned parallel to the telescope within 1 milli-
radian. The telescope is a folded Newtonian configuration with a 70 cm
diameter and a 2 meter focal length, and can be positioned to cover the
vertical hemisphere. The backscattered light is collected in the telescope,
transmitted to photomultipliers, detected and sent to the van for processing.
The detected signal is digitized every 10 nanoseconds and stored on
magnetic tape for data reduction in a large digital computer. The digitizer
also stores the signal return and presents it on an oscilloscope for real
time evaluation of the raw data. Auxiliary information such as telescope
position, date and time, laser power, and photomultiplier voltage are recorded
along with the data.
The lidar system is calibrated through the use of an 8 x 8 f^oot target
of known optical properties. These properties (reflectively, uniformity,
polarization properties) are measured in the laboratory before and after
each field trip.
2. Acoustic Sounder: The degree of pollutant trapping is strongly
influenced by the vertical temperature structure, particularly the existence,
height and strength of a temperature inversion layer. The vertical tempera-
ture structure was continuously monitored by an acoustic sounder of the type
described by Owens (1975). This unit also operates in a fashion similar to
radar. It emits a 100 watt, 100 millisecond sound pulse every 10 seconds in
a vertical beam. Variations in temperature scatter some of the sound back
into a five foot acoustic collector, where it is detected and presented on a
298
-------
24 GHz Radar
Retractable
Telescope
Data Processing
Equipment
Control Van
Figure 8.1. NOAA lidar system.
-------
facsimile chart. This information has proved very useful in the location of
inversion layers, so that the aircraft could be properly positioned for
maximum utility. Quantitative information regarding the strength of the
inversion was not obtained.
3. Insolation: An independent measure of the total aerosol content
between an observer and the sun is through the use of several insolation
(incoming solar radiation) devices. The method is to measure the total
incident radiation with an Eppley Model 2 pyranometer as well as direct
solar radiation at several wavelengths. The sun's radiation has been shown
to be constant (within 1%) over decades of time, so that variations can be
attributed to aerosols in the solar path. The effects of optically thin
clouds can be eliminated through the use of the lidar data, which is extremely
sensitive to clouds, being capable of detecting cirrus layers too thin to be
seen visually. The method fails when optically thick clouds block the sun.
Aerosol size distributions can be estimated by comparing the data obtained
at various wavelengths, since light scattering is dependent on the size-to-
wavelength ratio. These measurements were made with an Eppley Model 8
normal incidence pyrheliometer operating alternately unfiltered, and with
Schott OG-1, RG-2 and RG-8 filters. The pyrhel iometer was mounted on a sun-
tracker along with Barnes ITS radiometer to extend the measurements into the
8 to 12 urn region of the infrared.
Turbidity measurements were made using a Voltz turbidity meter operating
at the wavelengths .502 and .382 urn. Measurements were made three times
daily with some determinations being made several times per hour to test the
consistency of the measurements.
RECENT CHANGES
Following the first year's experience and findings from monitoring the
pregenerating plant atmosphere at Colstrip, considerable improvements and
additions to the monitoring capability were accomplished. Of particular
note was the "scaling-up" of airborne instrumentation to meet the requirements
of measuring the power plant plume diffusion, composition and chemical
reactions, effect on atmospheric optical properties, and consequences in
terms of possible precipitation attention.
Eleven parameters were measured from a Cessna 206 single-engine airplane.
These are: 1) Aitken nuclei with the continously operating €nvironment,
Inc. particle counter; 2) Cloud condensation nuclei with the Allee thermal
diffusion chamber-photographic method; 3) Ice nuclei by means of the membrane
filter method; 4), 5), and 6) Total oxides of nitrogen, nitric oxide, and
nitrogen dioxide concentrations with the Monitor Labs, Inc. chemiluminescent
instrument; 7) Ozone with the Monitor Labs, Inc. chemi1uminescent ozone
monitor; 8) Sulfur dioxide with the Theta Sensors, Inc. monitor; 9) Tempera-
ture, continuously recorded; 10) Relative humidity, continuously recorded;
and 11) Light scattering with the Meteorology Research, Inc. continuously
operating Model 1550 nephelometer. In addition, membrane filter collections
of atmospheric aerosols were done for subsequent analysis by Scanning Electron
Microscope (SEM) and Transmission Electron Microscope (TEM). The SEM with
energy dispersive x-ray analysis, yields the elemental composition of indi-
300
-------
vidual particles of diameter > 0.1 |jm, and for particle topography. The TEM
is utilized for the aerosol size distribution, which is crucial to calcula-
tions of possible atmospheric heating or cooling, and transmissivity.
The mobile laboratory was operated in the same location as for the
previous year's study. Parameters measured were: 1) Sulfur dioxide with the
Meloy Laboratories, Inc. flame photometric sulfur gas analyzer; 2) Atmospheric
light scattering with the Meteorology Research, Inc. nephelometer; 3) Ice
nuclei by the NCAR acoustic counter method (continuous); 4) Ice nuclei by the
membrane filter method; 5) temperature; 6) relative humidity; 7) wind speed;
and 8) wind direction. Membrane filters were also collected according to a
pre-determined schedule, for examination by SEM and TEM. Pilot balloon
releases and tracking were done once each hour for wind direction and velocity
aloft while the aircraft was operating.
The overall measurement capabilities are summarized in Tables 8.1 and
8.2.
DISCUSSION
LIDAR DATA
The system was on site for 30 days in the Spring (May 18 through June
16) and the Fall of 1975. The Spring was an unusually wet one, with some
rainfall occurring nearly every day. The Fall was more typical—mostly hot
and dry with occasional showers. The power plant was not in operation
during either period, so the data gathered will be used as a baseline for
future measurements.
Figure 8.2 is representative of the lidar data for the Spring period.
The signal begins at zero as the laser beam is initiated outside the tele-
scope's field of view. As the beam comes within the field of view the
signal increases until all of the beam is totally contained and is said to
have passed "crossover." Data recorded at heights lower than crossover are
not valid, since not all of the laser energy is used. This curve indicates
a nearly uniform atmosphere with an aerosol loading approximately 15 times a
theoretically clear atmosphere. The aircraft was operating 900 meters above
the surface when these lidar data was recorded and the optical backscatter
coefficient calculated from the particles collected in the membrane filter at
this altitude is also plotted.
Figure 8.3 shows a typical acoustic sounder record for the 1975 Spring
experiment. A weak inversion structure is seen operating from 0000 to about
0900 when thermal activity becomes the dominant feature. The dark bands from
1200 to 1400 are due to the aircraft operating overhead. Thermal activity
continues until 2000 when the evening temperature inversion reappears. The
Spring inversions, if present, were usually very weak, and would not have
played a significant role in the trapping of pollutants.
Figure 8.4 is representative of the 1976 Fall lidar data. It is essenti-
ally the same as the Spring data except for a sharp layer at a height of 500
meters due to aerosols trapped by a weak inversion layer. The corresponding
acoustic sounder record (Figure 8.5) barely shows the temperature inversion.
301
-------
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CO
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10
-7
COLSTRIP, MONTANA
DAY 151 MAY 31, 1975
-MEMBRANE FILTER
LIDAR
THEORETICAL CLEAR AIR
1000 2000
HEIGHT (M)
Figure 8.2. Optical backscattei—Spring 1975.
3000
-------
00
CO
O
0400
0800
1200
Time, (hrs)
1600
COLSTRIP, MONTANA
DAY 149 MAY 29, 1975
2000
0000
Figure 8.3. Acoustic sounder record—Spring 1975.
-------
10'
COLSTRIP, MONTANA
DAY 241 AUG. 29, 1975
10
-5
_
ct.
cc
-MEMBRANE FILTER
LIDAR
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10
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1000
2000
3000
HEIGHT (M)
Figure 8.4. Optical backscatter--Fal1 1975.
304
-------
GO
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CU3
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1200
Time, (hrs)
1600
Time, (hrs)
COLSTRIP, MONTANA
DAY 241 AUGUST 29, 1975
Figure 8.5. Acoustic sounder record—Fall 1975.
-------
In general, the Fall experiment generated much stronger inversion structures,
frequently containing multiple layers at several different altitudes (Figure
8.6).
During the Spring experiment the EPA-Las Vegas airborne lidar overflew
the site allowing the two systems to probe the same airspace at the same
time. This allowed the transfer of the calibration of the NOAA lidar to the
airborne system, which then flew a series of criss-crossed patterns in a box
30 miles on a side, centered about Colstrip. The airborne lidar data have
not been received.
The system was on site for 14 days in 1976. The weather was warm and
dry, but No. 1 power plant had been shut down for maintenance. However, No.
2 was operating intermittently during the last several days. Although there
has been insufficient time to analyze these recent data, some interesting
features have been noted. The 1976 data were taken using the dual-polari-
zation detector, to separate the stack signals from ordinary dust signals.
Figure 8.7 shows a tracing of an oscilloscope photograph of raw data in which
the system was simultaneously observing smoke from the power plant and dust
from the mines. The return from the smoke appears only in the "parallel"
channel, which means its plane of polarization is parallel to the plane of
polarization of the laser beam. This indicates the smoke particles are
spherical. The return from the dust, however, appears in both channels,
indicating that the dust particles are irregular. This is consistent with
the shape observed in an electron microscope of particles obtained in other
areas.
IN SITU DATA
Cloud and Ice Nuclei
The portion of the aerosol that has a direct bearing on the formation
and dissipation of clouds and precipitation are termed cloud and ice nuclei,
i.e., centers around which water droplets or ice crystals develop at low
supersaturations.
Before start-up of the Colstrip generating plant it was found that the
cloud condensation nuclei (CCN) concentrations at Colstrip were a large
fraction, 50 percent or even more for most of the time, of the total aerosol
population. The reason for this is the large proportion of chlorine- and
sulfur-containing particles that was found to exist in the Colstrip aerosol
(see Figures 8.8 and 8.9)._ Sulphur and chlorine usually are found in nature
in their anionic forms, SO^ and Cl , that form water soluble substances, and
hence are efficient as CCN.
Also prior to operation of the power generating plant, ice nuclei (IN)
concentrations were at least double those measured in most clean air regions.
The investigators believe this is a consequence of the high incidence of
heavy metals in the Colstrip aerosol. Zinc is present in many more particles
there than in aerosol samples from most other locations that the authors
have investigated. Some other metallic elements that occur unexpectedly
often are titanium, cadmium, chromium and nickel. Copper was found to be
306
-------
CO
o
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00
53
Time, (hrs)
1600
2000
Time, (hrs)
1200
COLSTRIP, MONTANA
DAY 231 AUGUST 19, 1975
Figure 8.6. Acoustic sounder record showing multiple inversion structure.
-------
OJ
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RANGE (KM)
Figure 8.7. Raw lidar return showing difference in depolarization between dust and stack effluent.
-------
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averaged over May and June 1975.
309
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COLSTRIP, MONTANA
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310
-------
occasionally present. Titanium dioxide and copper sulfide, in particular,
have been identified as active ice nucleating materials. In addition, the
elements aluminum and silicon have been identified in almost all of the
aerosols that were investigated. These elements are widespread in the
earth's crust in the form of the common clay mineral, Kaolinite, which is
known, in the aerosol form, to be a moderately active ice nucleant.
There exists one other class of atmospheric freezing nuclei. These are
biogenically derived and efficient at relatively warm (T > -5°C) temperatures.
While the presence of such nuclei is anticipated, there is no means in the
investigators' present capability for their detection.
Figures 8.10 and 8.11 are summaries of aerosol properties found during
the 1975 field investigations at Colstrip. The light scattering coefficient,
b ., is typical of rural western atmospheres; this is discussed further
bliow. The total particle population density (AN, or Aitken nuclei) is also
typical of clean atmospheres. The CCN and IN are discussed above.
A cursory look at the record from the airborne operation was done
following the May-June 1976 field investigation at Colstrip. From this it
is estimated that in the Colstrip power plant plume at short distances from
the stacks, the AN and CCN concentrations and b . are increased by factors
of 1.5 to 4. scat
AEROSOL OPTICAL PROPERTIES
Visible-light Optical Effects
Atmospheric aerosols affect the shortwave portion of solar radiation in
two ways: (1) They can reduce the amount of solar radiation reaching the
ground by scattering a significant portion backward into space, resulting in
surface cooling. (2) Absorption of a significant portion of solar radiation,
on the other hand, can lead to atmospheric warming. Which of these processes
predominates depends on the size and chemical constitution of the aerosol
particles.
Results from the analyses of sized and elemental composition (determining
the scattering portion of the particle refractive index) were used in a
computer program calculating the extinction, scattering and absorption
properties of the Colstrip aerosol. From the result it can be concluded
that the presently existing aerosol has a net cooling effect at surface
albedos less than 0.5, which is the case for the largest portion of the
year. Only during the winter months when the ground is snow-covered will
the situation be such that a warming effect can be anticipated.
The calculated scattering coefficient was verified with in situ measure-
ments using an integrating nephelometer.
Infrared Optical Effects
Aerosols absorb IR radiation in the water vapor window, thereby affecting
the accuracy of radiometric measurements of, e.g., temperature performed in
311
-------
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15 20 25 30 1 4 9 14
MAY —H JUNE I975
Figure 8.10. Aerosol properties—Spring 1975.
-------
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Figure 8.11. Aerosol properties--Fal1 1975.
-------
the 8-12 pm band from aircraft and satellites. At the same time they alter
the flux of terrestrial radiation into space, a phenomenon that directly
affects the earth's heat budget. The magnitude of this effect can be evalu-
ated by measuring with a radiation thermometer TJI situ at different altitudes
the brightness temperature of ground surfaces, which are in thermal equilib-
rium with the atmosphere. Varying the optical path length by a factor of
two, the investigators were unable to detect any effect of the Col strip
aerosol on the earth's equivalent black body temperature.
GASES
The background concentrations of nitric oxide, nitrogen dioxide and
sulfur dioxide outside of the plume are below instrument detectability. The
background concentration of ozone is around 30 parts per billion by volume.
Unit No. 2 of the Colstrip power plant was in intermittent operation
during June 1976. The following values of gaseous concentrations were
observed in the plume shortly after it exited into the atmosphere: NO and
N02 were both around 0.5 parts per million. S02 amounted to around one part
per million, and the ozone concentration was reduced to about 8 parts per
bi11 ion.
SOLAR IRRADIANCE DATA
The solar observing instruments were operated essentially continuously
during the 1975 Spring and Fall campaigns and during the recently completed
1976 Spring campaign.
The period August 28-September 15, 1975 was chosen for detailed analysis.
This period was characterized by mostly clear and partly cloudy weather which
permitted measurements of the aerosol-related attenuation almost every day.
The solar intensity data from the normal incidence pyrheliometer and the Volz
turbidity meter were corrected for sun-earth distance and normalized to
obtain the logarithmic (decadic) extinction coefficient with correction for
Rayleigh scattering and ozone absorption. Figure 8.12 shows hourly values of
pyrhelimometer decadic extinction for 09-17 hrs. It is seen that the extinc-
tion tends to increase during the morning hours and level off or decrease in
the later afternoon. Figure 8.13 shows this trend more clearly.
This is probably due to dust raised by mining activity'which would be
carried upward by thermal updrafts and drift to the vicinity of the observing
station. The acoustic sounder records (Figures 8.4 to 8.6) show thermal
plume activity beginning most mornings and lasting until near sundown.
Another interesting feature illustrated by Figure 8.12 is that the noon
extinction values tend to remain nearly constant for 3 to 4 days then become
changeable or switch to a new level. The cross-hatched areas indicate the
stable periods. This behavior is no doubt related to the movement of large
scale weather patterns and further study of this feature is planned.
Another way of looking at the variability of atmospheric attenuation is
by means of Langley plots (Figure 8.14), which display the logarithm of
intensity versus the airmass. The latter is a measure of the amount of air
314
-------
rot,STRIP, MONT-
.15 --
.12 --
r
y
r.
T
V
r
T
T
.1
H
.06--
.03- -
0.0
239
241
243
245
247
249
251
253
255
257
259
PAY
Figure 8.12. Hourly values of pyrhel iometer (0.3-2.8 urn) decadic extinction
(09-17 hrs) for day numbers 240-258 (Aug. 28-Sept. 15, 1975).
315
-------
DFCAHTC rXTU'CTTON COLSTFIF, MONT- 1<575
.15"
.12
.09
.06
.03
4-
06
12
TIMF
18
24
Figure 8.13.
Hourly values of pyrhel iometer (0.3-2.8 |jm) extinction vs time
of day for day numbers 240-258, 1975.
316
-------
.2
240
244
z
u
.1
.2
.3
.4
247
248
252-
251 243
5 10
AIRMASS
15
Figure 8.14. Superposed pyrhel iometer (3-28 urn) Langley plots for afternoons
during period Aug. 28-Sept. 15, 1975 (Day No. 240-258).
317
-------
through which the solar radiation must pass relative to what it would pass
through if the scan were directly overhead. The airmass is approximately
equal to sec6, where 6 is the angle between the vertical and the solar
direction, viewed from the observing site. Stable, horizontally uniform
atmospheric conditions would result in a straight line on a Langley plot with
slope related to the amount of vertical attenuation. Figure 8.14 therefore
illustrates both the day-to-day and the short-term variability of the atmos-
pheric attenuation due to aerosols. The relationship between decadic extinc-
tion as determined by the narrow band width Volz sun photometer and the
normal incidence pyrheliometer is shown in Figure 8.15. The minimum and
maximum values determined for each day by each instrument are plotted.
The extinctions obtained by the two instruments correlate very well on
the average. This supports the use of Volz turbidity data, which are rou-
tinely collected at some 50 stations in the United States.
The reduction of the direct solar irradiance, as measured by the pyrhe-
1iometer and Volz sun photometer and indicated by Figure 8.15, ranged from
about 7% to 25%. The total irradiance from the entire sky (i.e., the sum of
direct and diffuse radiation as measured by the pyranometer) varies generally
over a small range because the scattered diffuse radiation compensates for
the reduction in the direct component. In fact, under partly cloudy con-
ditions, with certain types of clouds near the solar direction, the sum of
direct and diffuse radiation can at times exceed the levels expected for a
clear day.
However, most aerosols absorb as well as scatter light with the result
that increased aerosol loading in the atmosphere results in the diminution of
total irradiation incident at ground level. This reduction in total irradia-
tion ranged from near zero to about 10% for the data of Figure 8.15.
The turbidity measured at Colstrip in the Fall of 1975 (average about
0.07) agrees closely with turbidity values published by Flowers et aj.
(1969) for Missoula, Montana, and Huron, South Dakota, which are, respec-
tively, 300-400 miles to the west and to the east of Colstrip. By contrast,
turbidity values published by Flowers et ajL for many of the observing
stations in the eastern half of the United States and for Los Angeles,
exceed 0.20, which means a diminution of the direct component of greater than
37%. Under the latter conditons a reduction of total irradiation by 15 to
20% does not seem unlikely.
Another factor which should be considered is the possibility that
introduction of additional aerosols may provide cloud condensation nuclei
leading to the formation of more clouds than would otherwise develop.
CONCLUSIONS
This section presents a summary of the conclusions at this stage of the
observations. The principal conclusions of this study will come only after
observations are made while the power plants are burning coal. During 1975
observations there was no plant operation. In Spring 1976, only sporadic
operation of Plant No. 2 occurred in a gas-fired mode.
318
-------
RANGE(MIN.andMAX.) OF DECADIC EXTINCTION
09:00-16:00 MDT AT COLSTRIP, MONTANA
BY VOLZ TURBIDITY METER
AT A=0.5nm
OF WIDEBAND 0.3-2.8Mm
PYRHELIOMETER
(RAYLEIGH SCATTERING LOSS AND OZONE
ABSORPTION SUBTRACTED IN BOTH CASES)
.151
.121
.091
O
z
H
X
U
O
5
<
o
u
.061
.031
239
243
247
251
255
259
DAY NO. 1975
Figure 8.15.
Minimum and maximum values of decadic extinction as determined
by pyrheliometer (dashed curve) and by Volz sun photometer
(solid curve) for day numbers 240-258, 1975.
319
-------
Observations in 1975 showed a very pristine atmosphere. Total particle
counts below 500 per liter were observed. The lidar returns showed very
little aerosol content over the whole range of the lidar. Occasional, mild
short-lived intrusions of dust from the mines were observed.
The acoustic sounder showed that many temperature inversions occur at
altitudes above 500 feet. In such cases there would be a tendency of stack
emissions (from 500 ft. stacks) to remain near ground level. Analysis of
acoustic sounder records is not complete.
The depolarization ratio of the lidar return is an important, although
not completely unambiguous, characterization index of the type of particles.
It is found that newly risen dust (from wind on explosions) has a large
depolarization ratio while stack emission has no discernible depolarization.
Solar radiation measurements showed marked differences due to aerosols.
It may be expected that significant changes in solar flux rates will occur
as the stack emissions increase.
LIST OF ABBREVIATIONS AND SYMBOLS
Lidar -- JJ_ght detection and ranging
IR -- infrared
SEM -- scanning electron microscope
TEM -- transmission electron microscope
CCN -- cloud condensation nuclei
IN -- ice nuclei
T -- temperature
AN -- Aitken nuclei
REFERENCES
Allee, P. A. "A description of the ESSA-APCL portable thermal diffusion
cloud chamber." Proceedings of the Second International Workshop on
Condensation and Ice Nuclei, 1974, Dept. of Atmos. Sciences, Colorado
State University, Fort Collins, CO. pp. 39-41.
Langer, G. , Rosinski, J. , and Edwards, C. P. "A continuous ice nucleus
counter." J. Appl. Met. 6, pp. 114-125 (1967).
Bigg, E. K. , Mossop, S. C., Mead, R. T., and Thorndike, N. S. C. "The mea-
surement of ice nucleus concentrations by means of 'Mi 11ipore.filters."
J. Appl. Met. 2, pp. 266-269 (1963).
Owens, E. J. "NOAA Mark VII Acoustic Echo Sounder." NOAA Technical Memo-
randum ERLWPL-12, U.S. Dept. of Commerce, Boulder, CO (1975).
Flowers, E.. C. , McCormick, R. A., and Kurfis, K. R. "Atmospheric turbidity
over the United States 1961-1966." J. Appl. Met. 8, pp. 955-962
(1969).
320
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FIELD AND LABORATORY
EXPERIMENTS
321
-------
SECTION 9
ZONAL AIR POLLUTION SYSTEM:
DESIGN AND PERFORMANCE
by
J.J. Lee and R.A. Lewis
INTRODUCTION
A major objective of the Montana Coal-fired Power Plant Project is the
generation of a protocol that will improve ability to assess the impact of
energy generation on the environment prior to critical land use decisions
concerning the initiation of power plant construction. The field experimental
component of this project is expected to provide data essential to the
construction of such a protocol. In particular, the investigators hope to
establish the specificity, thresholds, and temporal interrelations of the
effects of sulfur dioxide on well-defined components of a grassland ecosystem.
Also, by correlation of results of the field experiments with those from the
field studies near Colstrip, Montana, and perhaps elsewhere, they hope to
evaluate the site-specificity of these responses. Finally, during a five-year
investigation the investigators hope to generate predictive models that will
link short-term low threshold effects to those that occur only after prolonged
low level exposure.
The experiments are being conducted within two 27-acre grassland
exclosures in the Custer National Forest in southeastern Montana. Sulfur
dioxide (S02) fumigation of four one-acre plots within the first exclosure was
initiated in May 1975. Fumigation of the second set of four plots within a
second exclosure was started in April 1976. Livestock are excluded to protect
them from injury and also to protect equipment from damage.
The experiments are designed to test the effects of SO., upon biomass
dynamics (plant, arthropod, and small mammals), plant and animal community
structure, insect and fungal diseases of plants, pollination systems, lichens,
and upon a number of physiological and biochemical functions. Dominant plants
on the study plots are western wheatgrass (Agropyron smithi i), prairie
junegrass (Koeleria cristata). and Sandberg bluegrass (Poa secunda).
By using a gas delivery system developed for the project (Zonal Air
Pollution System--ZAPS), the investigators are able to maintain a selected
median concentration of sulfur dioxide on each plot during the entire growing
season (circa April through October). Continuous monitoring and control of gas
concentrations insure maintenance of the desired levels. Since the plots are
ecologically and physiographically similar, observed differences in effects
among plots can be attributed to S02 concentration. Fumigations are planned to
322
-------
continue for at least four growing seasons for the first set of plots, and at
least three for the second set.
A prototype system was designed and tested during September 1974. Results
indicated the feasibility of this type of control (Lee, Lewis and Body, 1975,
1976b; Lee and Lewis, 1976a). Predictions were made regarding the performance
characteristics of the system (see Table 9.4). Analysis of S02 data for the
1975 season confirmed these predictions (which were based upon modeling
solutions and tests of the prototype) and provided further evidence of the
adequacy of the system design.
DESIGN REQUIREMENTS
The ultimate design goal was to provide a system for the unambiguous
assessment of the sulfur dioxide impact on otherwise undisturbed grassland.
The designers approached this ideal by establishing realistic and appropriate
ecological and physical design criteria.
Disturbance of biota and of micro-climates by the system was to be
minimized. Effects on incident radiation, prey refuges, ground level
obstructions and pathways, temperature, humidity, wind and other features of
the micro-habitats were to be kept as small as possible. The area to be
stressed had to be large, on the spatial scales of the populations to be
sampled, to reduce edge effects and to ensure adequate population and sample
sizes. Furthermore, rational application of the system would require that
areas chosen for comparisons (i.e., "treatment" and "control" plots) be as
nearly uniform as possible in habitat, edaphic, and terrain features.
The logarithms of pollutant concentrations were to follow a normal
distribution as occurs in polluted areas (HEW, 1970). The distribution was to
be spatially uniform or nearly so, at least on a time average basis, and
concentrations were to be controllable for a range of selected averages.
Concentrations outside the plots and cross-fumigations among the plots were to
be minimized. Finally, cost, maintenance, and operation had to be reasonable.
THE GAS DELIVERY SYSTEM
The system that was designed and placed in operation for this
investigation generally meets the above criteria. Each gas delivery system
consists of a network of one inch aluminum pipes (schedule 40, alloy 6061-T6)
set parallel to the ground and supported at twenty foot intervals by five foot
pipes driven 2 1/2 feet into the ground (Figure 9.1). Release points (1/32
inch horizontal holes) are situated at 10 foot intervals, so that no location
within a plot is more than 18 feet from a source. A continuous flow of air
through the lines is provided by a helical compressor (Becker model SV 80-1).
On all but one of each set of four plots, S02 is bled into the air stream.
Equipment is housed in a heated shed with the temperature kept high enough so
that the vapor pressure of the S02 in the tanks is always above the desired
release pressure. Pressure regulators on the tanks and a flow controller
(Brooks model 8944) and flowmeter (Brooks model 1110) at the injection point
assure the desired flow rate of S02. Air pressure in the lines (about one pound
per square inch, gauge (psig)) varies by less than 5% across the system.
323
-------
M
T ]
280'.
\-
^r\
- ' 1
-
;
200'
M Central Monitoring Station
D Delivery Stations
G 6 KW Diesel Generator
WOO Three Conductor 0/0
Copper Wire
W2 Three Conductor 2
Aluminum Wire
W 2
WOO
IIOO'
C±1G
Figure 9.1. Schematic of the first set of Zonal Air Pollution Systems show-
ing the individual delivery systems and the common monitoring
and electrical systems. CONTROL, LOW, MEDIUM and HIGH indicate
relative S02 exposures on the experimental plots.
The only ground level obstructions within the plots are the supports.
These should cause minimum interference with animal movements, wTiile the pipe
network should negligibly influence the micro-climate. The relatively small
size of the plots precludes the study of large animals, although insects, other
arthropods and small rodents are included. The contiguity of plots allowed
selection for nearly uniform habitat conditions.
The four plots of each set are located along a line, with intervening
buffer zones to minimize interference between plots (Figure 9.2). To achieve
desired ambient concentrations during 1975 (Table 9.4), three of four plots of
the first set of plots received S02 at the constant rates of 1.5, 3.5, and 7.0
standard cubic feet per hour (SCFH) per plot. This implies that the S02
concentration in the delivery lines is approximately 1/2 1% (volume) for the
largest S02 rate; it is correspondingly lower for plots with lower S02 flow
324
-------
70'
70'
a
70'
28O'
C COMPRESSOR
S SULFUR DIOXIDE TANKS
H I KW HEATER
V VALVE
I" A! PIPE, 1/32" HOLES
EVERY 10*
I" Al PIPE, NO HOLES
A SAMPLERS
Figure 9.2. Schematic of a single Zonal Air Pollution treatment plot.
325
-------
rates. Concentrations on the plots varied with meteorological conditions. The
fourth plot received no direct input of S02 but did, however, occasionally
receive some S02 due to drift from other plots. Concentrations on this plot
exceed 5 pphm only 2% of the time.
The authors' application of a Gaussian dispersion model (Turner, 1969)
indicated that even under favorable dissipation conditions, concentrations
about 200 feet from a 40 pphm plot would remain below 5 pphm. The effects on
surrounding areas were thus expected to be minimal, and significant levels were
not expected to occur outside the study exclosure. The actual impacted area was
somewhat larger than expected, however, primarily due to channeling of the
pollutant by terrain features, and to a greater sensitivity of some of the
vegetation than was expected.
By utilizing many small, elevated, dilute point sources (over 250 per 1 1/4
acres), adequate dilution of S02 at ground level is insured and, in effect, an
area source is created. This prevents step-function changes in concentrations
in time and space ("hot spots"), except at very short distances from a source.
Testing of the system, as described below, confirmed expectations regarding the
nature of this distribution.
Knowledge of the physical parameters (pressure, density, orifice diameter)
has permitted M. Shirazi of The Corvallis Environmental Research Laboratory to
model certain features of the plume near a release point. The mixture of gases
is slightly heavier than the ambient air at the point of discharge. For the
analysis of this type of jet, it has been found that an important parameter is
the densimetric Froude number, defined by
F = U/VAp_ D g
P
This dimensionless number represents the ratio of inertia! to bouyant
forces. For the values of F greater than 40, the plume is neutrally bouyant;
i.e., the S02 can be considered to be a trace constituent whose density is
completely determined by the fate of its ambient air parcel. For the calcula-
tion of F, one needs the discharge velocity U, the jet diameter D, the ratio of
excess mass density Ap to the ambient density p, and the gravitational
acceleration constant g.
The parameters and calculations used to determine F are summarized in Table
9. la. The calculated discharge rate per jet is 1.8 x 10~3 ftVsec (about 27
ftVhr per plot), the discharge velocity is 330 ft/sec, and F is larger than
10,000. The S02 concentration is thus controlled by wind and turbulence, and
the S02 will not simply settle to earth and accumulate. This conclusion is, of
course, substantiated by the observation that S02 concentrations did not build
up during the growing season, and did not ever approach the concentrations in
the lines.
326
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TABLE 9.la. SUMMARY OF PARAMETERS AND CALCULATIONS USED TO ESTIMATE
DENSIMETRIC FROUDE NUMBER,
Mass Flow Through Jet:
W = 223.8 APo (C [(P_)2/k - (P_) (k+1)/k])1/2 ib/sec = 1.4 x 10-4 Ibs/sec
R =£ Po Po
I o
Where A = area of 1/32 in discharge jet, ft2
Po = pressure in line, 15.7 psi
P = pressure of ambient air, 14.7 psi
To = absolute temperature, 540° R
Cp = specific heat of air, constant p, .24 BTU/lb - °R
k - Cp/Cv where Cv is specific heat at constant volume, k = 1.4
R = universal gas constant divided by molecular wt. of air,
53.3 ft-lb/lb - °R
Volume Flow Through Jet:
V = WRTo = 1.8 x 10-3 ftVsec
Po
Discharge Velocity: U = V/A = 332 ft/sec
Excess Density of 1% (volume) Gas Mixture:
pmix (molecular wt) -x _gg x 28_9? + _Q1 x
1.012
pambient (molecular wt)ambient 28'97
Wp .012
The analyses of this type of problem indicate that jet dilution in still
air can be estimated by
0/0 = .32 X/D
o
where 0 is the total entrained ambient air within the plume at a distance X/D
jet diameters away from the point of discharge. Examples of dilution for
several distances are given in Table 9.1b. The actual observed dilutions
(Table 9.8) were at least 15 times greater than those predicted by this rough
model.
327
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TABLE 9.1b. PREDICTED DILUTION FACTORS AT VARIOUS DISTANCES FROM DISCHARGE
POINT.
Distance (ft) Dilution Factor
0.5 62
1.0 123
3.0 369
5.0 615
SYSTEM PERFORMANCE
A full size prototype of the system was constructed and employed to test
the feasibility of this experimental approach to the field during October 1974.
Results of these tests (Lee, Lewis and Body, 1975, 1976b; Lee and Lewis, 1976a)
demonstrated that concentrations across a plot are adequately controlled at the
low levels desired, but also suggested an improved design to minimize spatial
variation of concentrations. This new design (Figure 9.2) has been utilized in
all experimental gasing to date. Analysis of data for one entire growing
season indicates that the conclusions regarding the prototype were correct, and
that spatial variability is minimized by the new design.
S02 concentrations on the plots were monitored by recording the output
from a Meloy model SA160 sulfur analyzer operating in a logarithmic mode.
Samples were drawn through Teflon tubing by a time-share device (Adgo Co.) so
that each sample location was monitored at the rate of approximately 8 minutes
per hour.
Data on S02 concentrations were obtained one foot above ground level at
central location c (Figure 9.2) for each plot for the entire 1975 growing
season. Locations a and b (Figure 9.2) were also monitored for prolonged
periods for various plots. Location d was sampled occasionally; these data
have not yet been fully analyzed. In addition, specialized, short-term tests
of S02 distribution have been performed. These have been analyzed in terms of
temporal and spatial variations of S02. Comprehensive characterization of
horizontal and vertical distributions will continue during 1977.
TEMPORAL VARIATION OF S02
The frequency distributions of 8-minute medians (standard location c) are
shown in Figure 9.3, while Figures 9.4 and 9.5 show the frequency distributions
for 1-hour and 3-hour averages obtained from the 8-minute medians by
interpolation and averaging. All the distributions are approximately log-
normal, typical of pollutant frequency distributions (HEW, 1970; Larsen,
1969). Such distributions are characterized by the geometric mean (GM) and
standard geometric deviation (SGD). These parameters summarize the temporal
frequency of occurrence of concentrations, and thus represent a measure of
dose. Typical values for actual pollutant distributions are given in Table
9.2. Any area with similar GM and SGD can be said to have received a similar
dose.
328
-------
1 UU
E
f-
a.
CL
c
o
TD
E
c
E
CO
- 10
0
i—
a:
i-
z
LJ
O
z
o
o
CM
0
1
«
- D •
D •
- A a •
A a •
A D •
A D •
A D •
— O A O •
O A D •
O A D •
o A a •
O AD*
O A D ®
0 A D •
0 A 0 •
0 A D f>
- , , , o , A |D «
0.01 O.I I 10 50 90
CUMULATIVE FRACTION ABOVE X
99
Figure 9.3. Cumulative frequency distributions of 8-min. median S02 concen-
trations, 1975 season, o = plot A; A = plot B; n = plot C; •=
plot D.
329
-------
1 UU
E
a.
CL
C7>
o
a>
o
_c
oes
^JTRATION ( 1
o
UJ
o
z
0
o
-------
TABLE 9.2. GEOMETRIC MEANS (GM, pphm) AND STANDARD GEOMETRIC DEVIATIONS
(SGD) FOR SEVERAL URBAN AREAS. Five Minute Averages 1962-67
(source: 7)
City GM SGD
Chicago 1
Cincinnati
Denver
Los Angeles
Philadelphia
St. Louis
San Francisco
Washington
0.4
1.6
1.3
1.3
5.5
2.8
0.5
3.9
2.2
2.9
2.1
2.3
2.4
2.8
2.9
2.2
The values of these parameters
for the entire growing season, and
lower concentrations in October were
for the
in Table
probably
four plots are given in Table
9.3b for individual months.
due to frequent rain.
9.3a
The
Due to the logarithmic mode of the S02 monitor, all readings of zero on
the strip chart are recorded as concentrations of .9 pphm. The minimum re-
cordable GM is thus also .9 pphm. This effect was only important for plot A,
which was below .9 pphm more than 75% of the time.
In models of pollutant dispersion source strength appears as a normal-
ization constant, with the patterns of distribution determined by meteor-
ological conditions (Turner, 1969). The resultant low variability of SGD over
a range of GM's is evident for the urban areas summarized in Table 9.2 and for
the experimental plots in Table 9.3. The only dissimilar SGD is for the plot
which did not receive direct S02 input. The similarity of the SGD's in Tables
9.2 and 9.3 indicates that the fine-scale variability of S02 concentrations
throughout the study plots is realistic and reproducible.
Further evidence of the similarity of variability among study plots is
given in Figure 9.6. In this graph the S02 concentrations have all been nor-
malized to have the same GM as the highest concentation plot, plot D. This was
done by multiplying the S02 concentrations for plot "i" by the ratio GM (plot
D): GM (plot "i"). The coincidence of the resulting curves for plots
receiving direct S02 input shows that they differ in "amount" of S02 (GM) but
not in S02 "variability" (SGD). The curve for the fourth plot is affected by
the fact that the true GM is lower than the measured GM. The similarily of the
shape of this curve to the other curves is obvious in Figure 9.6.
The authors predicted S02 distribution parameters and peaks prior to the
introduction of S02 (Lee, Lewis and Body, 1976b). These are compared to the
observed values in Table 9.4. The seasonal 3-hour peaks are the most important
values from the standpoint of the present Federal Secondary Standard for S02
(Table 9.5). The differences between medians and geometric means are measures
of deviation from true log-normality.
331
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TABLE 9.3a. PARAMETERS OF S02 FREQUENCY DISTRIBUTIONS FOR 1975
GROWING SEASON.
For 8-min
Plot Medians
GM
A 1.0
B 2.2
C 3.8
D 6.8
SGD
1.5
2.4
2.7
2.7
For
1-hr
Averages
GM
1.0
2.1
3.6
6.4
SGD
1.5
2.4
2.7
2.6
For 3-hr
Averages
GM
1.0
2.2
3.9
6.2
SGD
1.6
2.2
2.5
2.6
GM: Geometric mean, pphm (minimum recordable is .9 pphm).
SGD: Standard geometric deviation.
TABLE 9.3b. GEOMETRIC MEANS AND STANDARD DEVIATIONS OF 8-MIN. MEDIAN S02
CONCENTRATIONS ON A MONTHLY BASIS.
Month
June
July
August
September
October
Plot
A
B
C
D
A
B
C
D
A
B
C
D
A
B
C
D
A
B
C
D
GM
(pphm)
1.18
2.54
4.40
7.52
0.96
2.43
4.47
7.59
1.02
2.31
3.54
6.38
1.02
2.32
4.42
8.37
0.97
1.43
2.32
4.02
SGD
1.50
2.05
2.40
2.75
1.38
2.29
2.49
2.86
1.62
2.44
2.56
2.52
1.59
2.61
2.89
2.69
1.45
2.17
2.63
2.46
332
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TABLE 9.4. COMPARISON OF PREDICTED1 AND OBSERVED S02 PARAMETERS, 8-MIN. AVERAGING TIME,
1975 GROWING SEASON.
Plot
Geometric
(pphm)
Mean
Predicted Observed
A
B
CO
co r
co L
D
0.0
2.0
5.0
10.0
1.0
2.2
3.8
6.8
Median SGD
(pphm)
Predicted Observed Predicted Observed
1
2.0 1.7 2.1 2.
5.0 3.4 2.1 2.
10.0 5.8 2.1 2.
5
4
7
7
Seasonal
3-hr. Peak
(pphm)
Predicted Observed
0
15-20
40-50
100-200
11
20
47
69
1 Predicted by Lee, Lewis, and Body, 1975.
-------
TABLE 9.5. MONTANA AND FEDERAL STANDARDS, pphm.
Montana
Federal Primary
Federal Secondary
1-hr. 3-hr. 24-hr.
25a 10b
14C
c
50°
anot to be exceeded more than once per four days (approx. 1%)
not to be exceeded more than 1% of time
cnot to be exceeded more than once per year
TABLE 9.6. PEAKS OBSERVED DURING 1975 SEASON, VARIOUS AVERAGING TIMES, pphm.
Plot
A
B
C
D
8-min
14
44
71
140
1-hr
11
32
54
89
3-hr
11
20
47
69
16-hr
3
13
24
33
TABLE 9.7.
PERCENT OF TIME
WERE EXCEEDED.
MONTANA (MT)
AND FEDERAL
(FED) STANDARD LEVELS
Plot
A
B
C
D
MT 1-hr
0
0.3a
3.8
10.2
MT 24-hr
0
1.2
27.5
59.3
Fed 3-hr
0
0
0
1.1
Fed 24-hr
0
0
10.0
34.6
Not frequent enough to constitute a violation of standard
334
-------
100
a.
a
c
o
6
c
'i
co
- 10
OL
\-
Z
UJ
<_>
•z.
o
o
- O A D •
- O Cb 9
O Ok •
O /X|]
-MO
4Q
o
o
A»
1
I
1
0.01 O.I I 10 50 90
CUMULATIVE FRACTION ABOVE X
99
Figure 9.6. Cumulative frequency distributions of 8-min. median S02 concen-
trations, normalized to same geometric mean, 1975 season. o =
plot A; A = plot B; n = plot C; • = plot D.
The peaks observed during the season for various averaging times are pre-
sented in Table 9.6, and compared with relevant State and Federal Standards in
Table 9.7. Sixteen-hour averages were used to approximate 24-hour averages.
The only plot on which the Federal Secondary Standard was exceeded was plot D,
while the Montana Standards were exceeded on all plots receiving S02 input.
The diurnal variation of S02 for a typical 24-hour period during the
summer is shown in Figure 9.7. S02 concentrations usually peaked during the
night and were low during the day. A different pattern was evident during late
summer and early autumn (Figure 9.8), with peaks occurring early in the morning
and late in the afternoon. This pattern is similar to that observed for the
prototype system (Lee, Lewis and Body, 1975).
The variation of the GM of 8-min. medians with time of day is shown in
Figures 9.9a-e for individual months and in Figure 9.9f for the entire season.
These graphs again show that concentrations were generally lowest at mid-day,
and that the pattern of S02 variation was similar across the plots.
335
-------
40
co
OJ
cn
Q.
Q.
C
o
O)
E
c
E
CO
o
or
UJ
o
z
o
o
-------
Diurnal patterns of pollutant S02 concentration from stationary point
sources and in urban environments are frequently similar to that produced by
this system (Holzworth, 1973; Le Quinio, 1973; Raynor et al_, 1974; Smith,
1968; Saito and Mizoguchi, 1973). Such variations in air pollutant concen-
trations are due to (a) variations in source strengths that in turn may result
from daily cycles in human activity (Garnett, 1973; Holzworth, 1973); (b)
variations in transport wind speeds and directions, atmospheric diffusion and
interactions (Cormier, 1974; Fukuoka, 1973; Garnett, 1973; Holzworth, 1973;
Lomaya and Tsintsadze, 1974; Martin, 1974; Smith, 1968). All of these vary
with weather and season (Balabuyev et a_l, 1973; Druilhet and Fontan, 1973;
Fukuoka, 1973; Sandig and Sandig, 1973). Atmospheric dilution is frequently
greatest during the day and least at night. This may result in one or more
daytime minimal and a nocturnal maximum in pollutant concentration (Holzworth,
1973).
JUNE
0
5.0 10.0 15.0 20.0 25.0
TIME OF DAY (hr)
Figure 9.9a. Variation of geometric means of S02 concentration with time of
day for the months June through October and for the entire 1975
season. o= plot A; A = plot B; o = plot C; * = plot D.
Horizontal line at 0.9 pphm represents the detection threshold;
all concentrations at or below this level were assigned a value
of 0.9 pphm for the purpose of computing geometric means.
337
-------
JULY
Q.
Q.
<
UJ
o
o:
l-
UJ
S
O
LU
10.0 15.0
TIME OF DAY (hr)
Figure 9.9b.
AUGUST
20.0
25.0
TIME OF DAY (hr)
25.0
-------
SEPTEMBER
E 20.0
Q.
Q.
UJ
o
ct:
h-
UJ
O
UJ
CD
15.0 -
0
5.0
10.0 15.0 20.0
TIME OF DAY (hr)
Figure 9.9d.
OCTOBER
5.0
10.0 15.0
TIME OF DAY (hr)
Figure 9.9e.
339
20.0
25.0
25.0
-------
WHOLE SEASON
20.0
25.0
TIME OF DAY (hr)
Figure 9.9f.
SPATIAL VARIATION OF S02
Tests on the prototype system showed that there were no "hot-spots" of S02
on the plot, and that S02 concentrations varied smoothly over the plot (Lee et
a]_. , 1976b). Investigations on the ZAPS plots during 1975 and 1976 have
provided further insight into the nature of the spatial variation of S02.
Numerous short-duration samples of S02 were taken near a S02 release
point. An example of such a series is given in Table 9.8. These indicated that
within three feet concentrations become comparable to locations halfway
between the delivery lines. This conclusion was upheld by data obtained over
five days during June 1976 for locations approximately six feet and 18 feet
from a release point (Table 9.9). Concentrations near the relea.se point were
typically 48% higher than those far from the release point.
Various points on the first set of four plots, and several points on the
second set, showed signs of excessive concentrations in the immediate vicinity
of release points. This was caused by jets being directed in a near-vertical,
rather than horizontal, direction. The data given in Table 9.9 is for a mis-
directed plume. It thus demonstrates the localness of the effect, and
indicates the maximum gradient to be expected. The misdirected jets will be
left as they are since the affected areas are not extensive enough to
invalidate the current experiments, and since the S02 gradients near these jets
provide unique opportunities for research.
340
-------
TABLE 9.8. VARIABILITY OF S02 CONCENTRATION IN THE VICINITY OF A S02 RELEASE
POINT NEAR LOCATION £.
Horizontal Distance
Downwind from Vertical Distance S02 Concentration
Release Point Below Release Point (10-min. median)
(cm) (cm) (ppnm)
0
0
25
25
25
25
50
100
200
300
30
15
65
30
15
0
0
0
0
0
4.3
18.2
3.1
3.1
185.0
5.5
4.2
14.6
5.1
6.1
Notes:
1. June 11, 1976.
2. Winds light and variable, 1-2 mph.
3. S02 concentration at standard location c was approximately 5 pphm.
4. Measurements are for consecutive 10 min. periods separated by 10 min.
periods during which the S02 monitor was zeroed with a S02 filter.
TABLE 9.9. COMPARISON OF S02 PARAMETERS 2 METERS AND 6.1 METERS FROM A
DELIVERY LINE. June 12-16, 1976.
Distance
From Line G.M. Median S.G.D.
(meters) (pphm) (pphm)
2.0 16.7 13.0 2.6
6.1 11.4 8.8 2.5
341
-------
A comparison of S02 concentrations at locations a, b, and c (Figure 9.2)
for plot B is given in Table 9.10. These are based on data obtained during most
of the 1975 season, and illustrate within-plot variability. The GM's agree to
within ± 5%, and the SGD's to within + 10%. The standard location (c) has an
intermediate GM, but exhibits somewhat more variability (highest SGD). This
location has the lowest 1-hr peak, an intermediate 3-hr peak, and the highest
16-hr peak. The S02 concentration at the standard location is typical of S02
concentrations across the plot.
TABLE 9.10. COMPARISON OF S02 CONCENTRATIONS ACROSS PLOT B.
Sample
Location
a
b
c
G.M.
(8-min medians)
(pphm)
2.0
2.2
2.1
S.G.D.
(8-min medians)
2.0
2.2
2.4
1-hr
Peak
(pphm)
43
33
32
3-hr
Peak
(pphm)
26
18
20
16-hr
Peak
(pphm)
8
8
13
SUMMARY AND CONCLUSIONS
The exposure system behaved in a predictable and realistic manner during
the 1975 season. Different S02 distributions were maintained on the four plots,
and these had the desired relationships to the Federal Secondary Standard for
S02. Maintenance was routine and not excessively time consuming. The system is
a useful and practical tool for determining the effects of S02 on naturally
growing ecosystems.
The behavior of the system is similar in several respects to a system that
was developed independently by a French research team (de Cormis, Bonte1 and
Tisne, 1975). In the French experiments, trees planted in a 2,000 m2 area have
been fumigated through 128 release points with S02 at an average concentration
of 3.4 pphm. The control consists of a similar area without a pollution
delivery system located 100 m from the fumigation plot. The French system uses
a S02 flow rate of approximately .85 SCFH, or about 80% that of the ZAPS after
allowance for differences in plot areas and S02 concentrations. The
concentration varies with meteorological conditions; high S02 concentrations
are, however, prevented by discontinuing fumigation when the-concentration
reaches a pre-determined value. They conclude that "The discontinuous nature of
pollution related to the effect of climatic conditions is not a handicap. On
the contrary, this phenomenon is quite similar to those encountered in certain
polluted sites which have provided the basis for this study."
ACKNOWLEDGEMENTS
The assistance of Denis Body in the construction, Ted Fletcher and Eric
Preston in the operation and on-line testing of the system, and of Brian
Satterfield in data gathering is gratefully acknowledged.
342
-------
REFERENCES
Balabuyev, A. G. , 0. V. Lomaya, and D. G. Tsintsadze. 1973. Annual and Diurnal
Course of the Concentration of Atmospheric Solid Aerosols Over a City
(Godovoy I sutochnyy Khod Kontsentratsii Atmosfernykh Aerozoley V
Gorodkikh Usloviyakh). Soobshch. Akad. Nauk Gruz. SSR, 69(3):585-589.
Cormier, R. V. 1974. The Nature and Variability of Integrated Boundary Layer
Winds. Preprint, American Meteorological Society, p. 244-249 (Presented
at the Conference on Weather Forecasting and Analysis, 5th, St. Louis,
March 4-7, 1974).
de Cormis, L. , J. Bonte1, and A. Tisne. 1975. Technique experimental permet-
tant 1'etude de 1'incidence sur la vegetation d'une pollution par le
dioxyde de soufie applique en permanence et a dose subnecrotique. Pollut.
Atmos. 17(16):103-107.
Druilhet, A. and J. Fontan. 1973. Determination of the Vertical Diffusion
Coefficients Between 0 and 100 m by Using Radon and THB. (Determination
des Coefficients de Diffusion Verticale entre 0 et 100 Male Aide do Rudun
et due THB) Boundary-layer Meteorol. 3:468-498.
Fukuoka, Y. 1973. Meteorological Study of Air Pollution: (1) The General and
Specific Cycles for Air Pollution. (Fukushima daigaku kyoikugakubu rika
hokoku). Sci. Rep. Fac. Educ. Fukushima Univ. 23:51-64.
Garnett, A. 1973. Emissions, Air Pollution and the Atmospheric Environment.
J. Inst. Fuel 46:39-45.
Health, Education and Welfare, Department of. 1970. Air Quality Criteria for
Sulfur Oxides, U.S. Govt. Printing Office. 178pp.
Holzworth, G. C. 1973. Variations of Meteorology, Pollutant Emissions, and Air
Quality. American Chemical Society, American Inst. of Aeronautics and
Astronautics, American Meteorological Society, U.S. Dept. of
Transportation, Environmental Protection Agency, Inst. of Electrical and
Electronic Engineers, Instrument Society of America, National Aeronautics
and Space Administration, and National Oceanographic and Atmospheric
Administration, 2nd Joint Conf. Sensing Environ. Pollut., Washington,
D.C. , p. 247-255.
Larsen, R. I. 1969. A new mathematical model of air pollutant concentration
averaging time and frequency. J. APCA. 19:24-30.
Lee, J. J., and R. A. Lewis. 1976a. Field experimental component: the bio-
environmental effects of sulfur dioxide. Iji R. A. Lewis and A. S. Lefohn,
ed., The Bioenvironmental Impact of a Coal-Fired Power Plant, First Interim
report, EPA-600/3-76-002.
Lee, J. J., R. A. Lewis, and D. E. Body. 1975. A field experimental system for
the evaluation of the bioenvironmental effects of sulfur dioxide. In W. S.
343
-------
Clark, ed. , The Fort Union Coal Symposium, MT. Academy of Science, E.
Montana College, Billings.
. 1976b. The Field experimental component: evaluation of the
Zonal Air Pollution System. I_n. R. A. Lewis, N. R. Glass, and A. S. Lefohn,
ed. , The Bioenvironmental Impact of a Coal-Fired Power Plant, Second
Interim Report, EPA-600/3-76-013.
Lomaya, 0. V. and D. G. Tsintsadze. 1974. Analysis of the Diurnal Course of Air
Pollution. (Analize sutochnogo khoda zagryazneiya Vozdukha). Soobsch.
Akad. Nauk. Gruz SSR. 73:329-332.
Martin, D. E. 1974. Some Air Pollution Climatologies of the St. Louis Urban
Complex. Preprint, American Meteorological Society, p. 180-182.
Presented at the conference on Weather Forecasting and Analysis, 5th, St.
Louis, MO, March 4-7, 1974.
Le Quinio, R. 1973. Concentrations sur une Heure de pollutants a des emissions
ponctuelles pres du sol-presentation probabiliste. Atmos. Environ.
7(4):423-428.
Raynor, G. S. , M. E. Smith, and I. A. Singer. 1974. Temporal and Spatial
Variation in Sulfur Dioxide Concentrations on Suburban Long Island, New
York. J. Air Poll. Contr. Assoc. 24:586-590.
Smith, M. E. 1968. The Influence of Atmospheric Dispersion on the Exposure of
Plants to Airborne Pollutants. Phytopath. 58:1018-1088.
Saito, K. and T. Mizoguchi. 1973. Measurement of Sulfur Oxides and Floating
Dust by the Air Contamination Automatic Measuring Recorder. (Taikiosen
jido sokutei kirokukei ni yoru 10 sankabutsu oyobi Fuyu bijin no sokutei ni
tsuite ken eisei nura kenkyusho nenpo. 6:145-147.
Sandig, R. and R. Sandig. 1973. (Zur Edv-gestutzten Bestimmung der Schwef-
eldioxid-immissionen an einer Stationaeren Messstelle im Stadtzentrum von
Zwickau). Z. Ges. Hyg. Grenzgebite (Berlin). 19:890-896.
Turner, D. B. 1969. Workbook of Atmospheric Dispersion Estimates. U.S. Dept.
of Health, Education and Welfare. 84 pp.
344
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SECTION 10
FIRST-YEAR EFFECTS OF CONTROLLED SULFUR DIOXIDE FUMIGATION ON A
MIXED-GRASS PRAIRIE ECOSYSTEM
by
J. L. Dodd, W. K. Lauenroth, R. K. Heitschmidt and J. W. Leetham
INTRODUCTION
At the beginning of the Col strip Coal-fired Power Plant project it was
concluded that sulfur dioxide (S02) was probably the single most important gas
of the several effluents expected from the power plant. A field experiment was
designed to expose four 0.5 ha grassland study sites to different ambient
concentrations of sulfur dioxide on a continuous basis throughout the growing
season. The objectives of the experiment were to determine the effects of 2, 5
and 10 pphm monthly median sulfur dioxide concentrations on key structural and
functional attributes of the grassland. These concentrations were selected to
bracket Federal and State of Montana primary and secondary air quality
standards.
The fumigation experiment is located on a grassland site in the Custer
National Forest 85 km southeast of Colstrip. At this distance it is presumed
to be out of the influence of the Colstrip plume. The site is located at an
elevation of about 1,200 m on the divide between the Powder River and the
Tongue River drainage basins.
Soils of the study site are Farland silt loam. These are well developed
soils and on the site, have an A-horizon about 25 cm thick over a B-horizon that
is about 70 cm thick. The climate in this part of the Great Plains is
continental with about 75% of the precipitation occurring during the growing
season. The annual rainfall at the study site is about 350-400 mm.
The vegetation of the site is typical of the northern mixed-grass prairie
in that it is dominated by cool season grasses, mainly western wheatgrass and
junegrass, and has smaller quantities of other grasses and a variety of forbs
and half-shrubs.
Data on various abiotic variables including air and soil temperatures,
relative humidity, wind velocity, soil water content, and precipitation were
collected for most of the growing season. Growing conditions in the early part
of the 1975 season were very favorable with May and June each receiving over
100 mm of precipitation. This is probably well above normal for the area. Soil
345
-------
water was near field capacity until about the first of July and then decreased
rapidly and remained low throughout the remainder of the season.
PHENOLOGY
Previous investigations concerning the effects of S02 upon the primary
producer component of an ecosystem have virtually ignored reproductive
processes. The possible effects of air pollutants on pollination, flower set,
seed development, and seed dispersal should not be overlooked. Any changes in
the time of occurrence of these events as a result of exposure to air
pollutants may significantly alter the plant's reproductive capabilities.
The objective of the phenology study was to identify any major differences
between treatments in the phenological development of the major plant species.
Fourteen species were observed on a weekly time interval throughout the
growing season (Table 10.1). As can be seen, a majority studied were cool
season species and, for a considerable portion of these species, phenological
development occurred prior to initiation of the continuous fumigations in late
May.
TABLE 10.1. SCIENTIFIC AND COMMON NAMES OF THE 14 MAJOR PLANT SPECIES
SELECTED FOR PHENOLOGICAL STUDY
Scientific Name Common Name
Cool season grasses
Agropyron smithii Western wheatgrass
Bromus japonicus Japanese brome
Koeleria cristata Prairie junegrass
Poa secunda Kentucky bluegrass
Stipa comata Needle-and-thread
Warm season grasses
Aristida longiseta Red-three-awn
Bouteloua gracilis Blue gramma
Warm season forbs
Antennaria rosea Rose pussytoes
Psoralea argophylla Silverleaf scurf pea
Cool season forbs
Achi1 lea mi 11ifolium Western yarrow
Sphaeralcea coccinea Scarlet globe mallow
Taraxacum officinale Common dandelion
Tragopogon dubius Yellow salsify
Half shrubs
Artemisia frigida Fringed sagewort
346
-------
A 14-stage phenological classification (Table 10.4 in Section I) was
utilized to describe the observed phenological status of the selected species
within each treatment on each observation date. The recorded status reflected
the mean of all plants of the species within a treatment.
Statistical analyses indicated no significant differences between
treatments in the time of occurrence of major phenological events. However, a
definite trend was noted indicating an advancement in phenological development
with increasing S02 concentration. This trend was evidenced by 10 of the 14
species observed (Appendix).
An example of a species displaying this trend is false salsify, a cool
season forb (Figure 10.1). The low and high treatments reached the mature
floral bud stage, stage 9, one week prior to the control and medium treatment
while all three treatments reached the open flower stage, stage 9, two weeks
prior to the control. In addition, the medium and high concentrations began
seed dispersal approximately one week prior to the control and low treatments.
This trend was reflected in all groups except the half-shrubs in which fringed
sagewort was the only species observed.
Although it was concluded from statistical analyses that no major
differences occurred in the timing of phenological events as a result of S02
fumigation, the trend toward advancement with increasing S02 concentration
suggests minor differences may have occurred. Since observations were only
conducted approximately every 6 to 7 days, treatment differences of only 2 to 3
days would probably not have been revealed except on every third or fourth
observation date. Furthermore, since continuous fumigation was not initiated
until after a considerable portion of the phenological progression of the cool
season species had occurred, a more dramatic effect might be expected if
fumigation were initiated at the beginning of the growing season.
VISIBLE INJURY
Visible injury to plant tissue resulting from S02 exposure has been the
subject of numerous studies since the early 1930's (Hill and Thomas, 1933;
Thomas and Hill, 1935; Nat. Res. Council Con., 1939). The earliest studies
primarily focused on describing characteristic patterns of visible injury as
exemplified by different species and the effect of this injury on yield in
various agronomic crops (Briesley £t al., 1950, 1959). More recently, emphasis
has been placed on determining the "threshold concentration" inducing visible
injury (Thomas, 1956; Daines, 1968). These "threshold concentrations" have been
found to vary with changes in concentration, duration of exposure, and abiotic
variables such as temperature, relative humidity, and light (Daines, 1968).
However, a large majority of these studies have been conducted under controlled
laboratory conditions and normally evaluated only acute effect of high
concentration short-term exposures. Field-observed chronic visible injury
induced by low concentration long-term exposures has not previously been
reported for native range species.
The objective of the following research was to identify and describe any
visible injury incurred by western wheatgrass as a result of S02 fumigation.
347
-------
CO
-p*
co
14-,
icH
e'
o
o
o 6-
ID
C.
CL
0
EJ Control
tD Low
[HI Medium
m High
False Salsify
26 May 3Jun 9 Jun ISJun 22Jun 3OJun 7Jul ISJul 21 Jul 29Jul 4 Aug
Figure 10.1. Phenoloqical development of false salsify (Traqopogon dubius) for Taylor Creek, 1975.
-------
Western wheatgrass was selected for this facet of the study since it is the
dominant species within the fumigation plots and because its morphological
characteristics are such that field collection, mounting, and visible injury
determinations are simplified.
Four plants were collected outside the perimeter of each harvest quadrat on
the June, July, August, and September sample dates. After the quadrat was
randomly located, the four closest western wheatgrass plants to the corners of
the quadrat were clipped at ground level. Thus, 20 plants were collected per
replication and 40 plants per treatment.
After clipping, the leaves were removed and mounted in order of age between
two strips of clear adhesive acetate. Each leaf's total surface area and total
injured area was then recorded from visual determinations using a guided
overlay. Visible injury was defined to include any portion of the leaf
exhibiting necrotic condition, including normal leaf senescence. The total
surface of missing leaves was considered injured.
An increase in the degree of leaf senescence has previously been reported
as the only visible injury detectable in perennial ryegrass plants when exposed
throughout a growing season to low concentrations of S02 (Bell and Clough,
1973). Examination of the leaf surfaces of our sampled plants also revealed no
distinguishable pattern of visible injury other than an increase in leaf
senescence. Consequently, statistical analyses were run in an attempt to detect
differences between the percent of leaf surface necrosis in the control plants
and the fumigated plants. Visible injury, resulting from exposure to S02, as
used here, is defined as the necrotic portion of the leaf surface exceeding
normal senescence as reflected by comparison of the fumigated plants with the
control plants. Leaf injury means for western wheatgrass are presented in
Figures 10.2 (by data treatment and leaf age) and 10.3 (by leaf age and
treatment date). The Analysis of Variance (ANOVA) utilized to test the
differences between treatments across the four dates indicated a significant
difference between the LOW concentration and the HIGH concentration with no
differences between all other means (Table 10.2). As expected, a significant
difference (P < 0.05) in total injury between the four dates was also found, but
this is a reflection of the normal aging process. Since the last sample date was
very late in the growing season, it was felt that normal fall senescence might
be masking the effects of the S02. Therefore, an ANOVA was run encompassing
data only from the first three sample dates. This analysis indicated
significant treatment differences between the CONTROL and the LOW treatment when
compared to the HIGH treatment and between the LOW and the MEDIUM treatments
(Table 10.3). It is felt that the LOW treatment exhibited a lower percent of
leaf necrosis than the control because of a significantly higher number of
leaves per sampled plant on the sample date of 6 August. Thus, with a larger
proportion of the total leaf surface composed of younger tissue, a lower mean
percent of injury was reflected by each plant. It has been reported that
middle-aged leaves are more susceptible to S02 damage than either older or
younger leaves (Guderian and Van Haut, 1968). In an attempt to examine this
phenomenon, an ANOVA of only the first four leaves was undertaken.
349
-------
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Figure 10.2. Seasonal change in percentage of necrotic leaf surface for first, second, third, and fourth
leaves of western wheatgrass, Taylor Creek, 1975.
-------
lOO-i
75-
•E 50-
CO
CJ1
c
0)
O
25-
0
Leaf I
Leaf 2
Leaf 3
Leaf 4
Figure 10.3.
Effect of S02 treatments on leaf injury (percentage of leaf surface necrotic averaged
across three dates—June, July, and August) by leaf age, Taylor Creek, 1975.
-------
TABLE 10.2. TREATMENT MEANS FOR PERCENT LEAF INJURY SUSTAINED BY WESTERN
WHEATGRASS OVER FOUR SAMPLE DATES. Means underscored by
same line are not significantly different (P < 0.05).
LOW
38.91
CONTROL
42.14
MEDIUM
44.16
HIGH
48.61
TABLE 10.3. TREATMENT MEANS FOR PERCENT LEAF INJURY SUSTAINED BY WESTERN
WHEATGRASS OVER THREE SAMPLES DATES. Means underscored by
same line not significantly different (P < 0.05)
LOW
26.43
CONTROL
29.74
MEDIUM
34.63
HIGH
38.09
Both the 3-date and the 4-date ANOVA indicated a significant difference
between all leaves in the percent of the total leaf surface necrotic. A
significant increase in leaf necrosis occurred with increasing leaf age, but
this is as expected since normal leaf senescence increases with leaf age.
However, an important difference in the two analyses was noted in that no
significant treatment by leaf interactions was found to be significant in the
3-date analyses. Tukey's Q values (P = .05) indicated no significant
differences between the percent injury exhibited by the four treatments' first
leaves, but a significant difference occurred between the CONTROL and the two
HIGH concentrations for the second, third, and fourth leaves. The significance
of this interaction cannot be overemphasized because it clearly establishes
that an increase in leaf senescence did occur as a result of the S02
fumigation.
Analyses of the number of leaves lost per plant, the age of the lost leaf,
and the age of the plant losing the leaf indicated no treatment differences.
In general, the oldest leaves of the plants were lost first in all treatments.
In summarizing the above findings, three conclusions have been drawn
concerning the effects of the S02 fumigation treatments on foliar injury:
1. Chronic visible injury for western wheatgrass is primarily expressed
by an increase in leaf senescence without specific pattern.
2. A significant increase in visible injury occurred with increasing S02
concentrations.
3. A significantly greater amount of injury was incurred by the older
leaves than by the younger leaves.
352
-------
It should be noted that the visible injury conclusions are based on
statistically significant differences between the CONTROL and the MEDIUM and
HIGH treatments and no differences were found to occur between the CONTROL and
the LOW treatment.
The significance of these conclusions, as they relate to a range of
ecosystems, remains to be seen. Guderian and Schoenbeck (1971) attempted to
distinguish between the two synonyms "damage" and "injury" as they pertain to
plant responses resulting from the action of air pollutants. They suggest that
"injury" should be considered to include all plant responses to air pollutants
while "damage" should only refer to those responses which significantly alter
the desired use of the plant. After one season of data collections and
analyses, the investigators feel the injury has been detected as reflected by
the increase in leaf senescence with increasing S02 concentration. However, it
remains to be seen whether or not damage has occurred.
PLANT BIOMASS DYNAMICS AND NET PRIMARY PRODUCTIVITY
The objective of this part of the study was to characterize the effects of
three levels of sulfur dioxide fumigation on above- and belowground plant
biomass dynamics and net primary productivity.
Aboveground biomass was sampled by the harvest method and separated in the
field by species and into three categories: live, recent dead, and old dead.
Live and recent dead represent growth during the current growing season and old
dead is biomass remaining from previous growing seasons. On each of six sample
dates, ten randomly located, circular 0.5 m2 quadrats were harvested from each
treatment.
Total biomass accumulation by all species was similar with no significant
differences among treatments at any time during the season (Figure 10.4). The
average growth rate from 15 April to 15 July was 1.4 g/m2/day.
Fumigation was initiated on 1 June and had no measurable effect on biomass
accumulation. The point at which we measured a change in the rate of growth was
6 August. This decrease in growth rate appears to be related to a rapid decline
in soil water between 15 July and 6 August rather than a response to the
fumigation treatments.
The two most important functional groups contributing to total biomass are
cool season grasses and cool season forbs. Biomass accumulation by both of
these groups was more variable among treatments than total growth, but no
significant differences or explainable trends were evident (Figure 10.5).
Average growth rate of cool season grasses from 15 April to 15 July was 1.1
g/m2/day. At the time that fumigation was begun, approximately one-third of
the cool season grass growth had occurred. Cool season forb growth was
greatest for the HIGH concentration treatment and lowest for the LOW treatment
with the control falling approximately midway between the two.
Stepping down to a finer level of resolution one can see the responses of
the two most important species in the cool season grass group. The first of
these is western wheatgrass (Figure 10.6). Western wheatgrass is an important
353
-------
150 -,
GO
in
IOO -
E
\
o>
(O
to
O
e
o
00
50 -
0 -
25.E.
Control
Low
— — - Medium
High
Apr
May ' Jun ' Jul ' Aug ' Sept'
Figure 10.4. Seasonal change in current year1s production for all species combined, Taylor Creek, 1975.
-------
150 -i
CO
CJ1
en
100 -
CO
o
o
GO
50 -
Cool Season Grass
Control
Low
— — — Medium
High
Cool Season Forb
2S.E.
2S.E.
Apr
May
I Aug
Sept
Figure 10.5. Seasonal changes in current year's production for cool season grasses and forbs, Taylor
Creek, 1975.
-------
GJ
cn
CT)
IOO -i
CM
o>
10
(/>
CJ
I
80 -
60 -
20 -
0
Western Wheatgrass Growth-—
Control
Low
Medium
High
25-E.
western Wheatgrass Dead
^^—- I2SE
2SE
May
Jun
Jul
Aug
Sept
Figure 10.6. Seasonal changes in current year's production and previous year's dead for western wheat-
grass, Taylor Creek, 1975.
-------
dominant over much of the northern Great Plains and is probably the most
important species to the livestock economy of the region. The growth rate of
western wheatgrass averaged 0.6 g/m2/day from 15 April to 15 July and 0.5
g/m2/day for the entire period of measurement.
While these data do not demonstrate treatment effects, they do represent a
solid characterization of the dynamics of growth and disappearance to dead
material during the 1975 growing season. Additionally the May sample of live
and recent dead biomass and the April and May samples of old dead indicate high
degree homogeneity of western wheatgrass biomass across treatments. The
importance of this homogeneity relates first to the question of whether
significant effects of fumigation on the growth of western wheatgrass occurred
during 1975 that were offset by initial differences in populations among
treatments. These data indicate that this was not the case. Secondly, the
homogeneity of western wheatgrass biomass during the first year of treatment
will strengthen our conclusions about differences should they occur during the
second or third years of treatment.
Prairie junegrass is the second important cool season grass on the
experimental plots (Figure 10.7). Several of the differences in prairie
junegrass biomass accumulation among treatments are significant although these
differences have not been explained in terms of treatment effects. The data
for old dead biomass on the first sample date indicate inherent variability in
prairie junegrass populations among treatments and at this time differences in
prairie junegrass biomass accumulation are attributed to this variability.
Belowground biomass was sampled to a depth of 20 cm by the harvest method.
Included in these samples were crowns, rhizomes, and roots. Crowns and
rhizomes were separated by hand and represent primarily live tissue while root
biomass includes all belowground organic material greater than 0.5 mm2.
Total root and rhizome biomass were not significantly different among
treatments on any of the sample dates (Figure 10.8). The most perplexing
aspect of these data are the two different trends. All of the fumigated
treatments represent one trend while the control exhibits a different trend.
There is evidence from the literature that exposure to ozone may reduce root
reserves. The action of ozone and S02 may be similar (Tingey, 1974) and
therefore explain these trends, but at this point conclusions based on one
year's data cannot be made. Crown biomass was similar among treatments and
essentially constant at approximately 60 g/m2.
Total roots and rhizomes were separated into two depth increments of roots
and rhizomes alone (Figure 10.9). It is evident that the major differences in
belowground biomass occurred in the 0-10 cm layer and did not occur until after
fumigation was initiated. Trends in root biomass were similar among treatments
between the April and May sample dates and different for all other dates.
Again, these differences cannot be satisfactorily explained with a single
year1s data.
Root biomass in the 10-20 cm layer was essentially constant throughout the
growing season at approximately 100 g/m2. Rhizomes also exhibited only minor
fluctuations and averaged 25 g/m2.
357
-------
CO
en
CO
75 -,
5O -
CM
O
E
o
CD
25 ~
0 -
Control
Low
— — — Medium
High
Prairie Junegrass Growth
Prairie Junegrass Dead
J2S.E.
2S.EJ
1 Jun
Jul
"I Aug I Sept
Figure 10.7. Seasonal change in current year's production and previous year's dead for prairie June-
grass, Taylor Creek, 1975.
-------
9OO
S!
^ 70O
D
CM
E
en
O
O
CD
en
o
-d
O)
I
c
6
s_
i1
_o
CD
CD
5OO -
200 -
Total Roots Rhizomes
2 S.E.
Control
Low
— — — - Medium
— • High
I
Crowns
2 S.E.
Apr
May
Jun
Jul
Aug
Sept
Figure 10.8. Seasonal changes in biomass of crowns and roots plus rhizomes (0.020 cm), Taylor Creek,
1975.
-------
aoo ~i
CO
er>
o
o>
o>
I
6OO -]
o
o
bo
8.
.o 40O
\_
CD
TD
C
•
o
0}
CD
Control
Low
Medium
High
Roots 0-IOcm
2S.E.
2OO
O
^— "= -^T-J =-_
Apr 1 M
i.-=.-^" —--="•"
ay ' Jun
1 Jul
=^^_- -. • •— —
1 Aug
1 Sept
I
1
Roots IO-2Ocm
2S.E.
Rhizomes
I 25.E.
Figure 10.9. Seasonal change in biomass of roots at 0-10 cm and 10-20-cm depths.
-------
Since the results of the belowground biomass sampling are still in
question estimates of belowground net accumulation have not been made.
Aboveground net primary productivity was calculated by summing peak live plus
recent dead biomass by functional groups (Figure 10.10). At first glance it
appears that primary productivity increased with increasing S02 concentration.
More careful observation reveals that the higher values for the MEDIUM and HIGH
treatments were the result of composition differences. Both of these
treatments had measurable contributions of half shrubs while they were only
sampled in trace amounts for the CONTROL and LOW treatments. Comparison of
aboveground productivity excluding half shrubs shows no major differences
among treatments.
In summarizing these results it is emphasized that no significant
differences in seasonal biomass dynamics or net primary productivity were found
as a result of one year of sulfur dioxide fumigation. There are several
reasons why this was not an unexpected result. First, fumigation was not begun
until approximately one-third of the total plant growth had occurred. Second,
the major primary producers on the experimental plots are grasses and as a
group they are generally considered to be less sensitive to S02 than other
plants (Hill et a!. , 1974). Finally since the sample methods only yield
interpretable data on the more abundant species, many significant changes could
have occurred in populations of species comprising the forb group that the
investigators could not detect. In reference to this last point, sample
techniques have been modified for the 1976 growing season and a greater degree
of precision for these data is anticipated.
LITTER DYNAMICS
As shown in Table 10.4 the influence of season and treatment on litter
standing crop estimates are unclear. It appears that litter increased between
April and May and again between July and August. Differences in litter
standing crop resulting from the S02 stress are not consistent and are probably
not statistically significant.
SOIL RESPIRATION
Soil respiration was estimated utilizing four samples per replication
with two replications per treatment on four sample dates (Table 10.5).
Difficulties encountered in the laboratory processing of samples prevented
early season sampling. The higher rates encountered on 15 August at Taylor
Creek and 18 August at Colstrip are attributed to rainfall events immediately
preceding the sampling period.
361
-------
Aboveground Net Primary Productivity
20O-
oo
01
no
e
\
o»
co
O
E
o
im
!OO-
Cool Season Grasses
Cool Season Forbs
Warm Season Grasses
Warm Season Forbs
Half- Shrubs
Control
Low
Medium
Treatment
High
Figure 10.10.
Aboveground net primary productivity for Taylor Creek study sites, 1975.
by functional groups represent peaks of current year's production.)
(Contributions
-------
TABLE 10.4. INTRASEASONAL DYNAMICS OF LITTER STANDING CROP (x ± SE, Oven-dry Ash-free g/m2) FOR TAYLOR
CREEK SITES (A = CONTROL, B = LOW, C = MEDIUM, D = HIGH).
Treatment
co A
CO
B
C
D
Average
April 20
112
112
135
39
± 6
± 21
± 16
+ 7
100
May
102 ±
174 ±
168 ±
179 +
156
5
16
21
24
23
June 1
154 ±
137 ±
178 ±
124 +
151
5
8
14
17
12
July 13
123 ± 5
155 ± 17
155 ± 18
154 ± 16
147
August
172 ±
192 ±
193 ±
189 ±
187
13
17
15
17
22
September 18
145
149
193
100
± 20
± 15
± 17
+ 33
175
Season
Average
138
153
170
148
-------
TABLE 10 5 SOIL RESPIRATION TREATMENT MEANS (mg C02/m2/day) AND
STANDARD ERROR OF ESTIMATE FOR THE FOUR TAYLOR CREEK
TREATMENTS ON FOUR DATES, 1975 (A - CONTROL, B = LOW,
C = MEDIUM, D = HIGH).
Date
2 August
8 August
15 August-7'
27 August
5.
4.
6.
5.
A
12 ±
38 ±
92 ±
97 ±
.28
.19
.20
.56
5.
4.
6.
4.
B
43 ±
46 ±
43 ±
72 ±
.21
.16
.12
.20
5.
4.
6.
4.
C
72 ±
00 ±
69 ±
01 ±
.23
.38
.29
.34
D
5.79 ±
4.10 ±
6.81 ±
4.18 ±
.50
.31
.26
.52
- Mean of one replication.
Although final analysis is not complete the 27 August sample at Taylor
Creek indicates a significant difference exists between the control (A) and the
S02 treatments. If this difference is found to be real, two obvious
explanations exist: (1) reduction in microbial activity because of S02
treatment; and (2) a reduction in root respiration because of S02 treatment.
More intensive sampling is scheduled for 1976 in anticipation of ascertaining
the seasonal dynamics of all sites and treatment differences at Taylor Creek.
LITTER BAG DECOMPOSITION
The rate of decomposition of native litter was investigated utilizing
approximately 5 g of dead western wheatgrass, Agropyron smithii, placed in a
mesh bag on the soil surface and retrieved at various time throughout the
growing season. The means of the percentage of organic matter decomposed from
10 samples per replicate (two replicates per treatment) are presented in Figure
10.11.
Although statistical analysis has not been completed, two trends are noted
from the data. As expected, the maximum rate of decomposition within all
treatments occurred in the early part of the growing season. This is
attributed to the higher moisture conditions and, thus, favorabje for decomp-
osition. The second trend noted is the higher amount of decomposition within
the control when compared to the S02 treatments. This trend suggests a
reduction in microbial activity may have occurred because of the S02 fumiga-
tion. However, since the differences between treatments are essentially the
same on all dates it is questionable whether the differences can be attributed
to S02 fumigation. Since the fumigations were not initiated until 1 June, the
differences found on 10 June may be attributable to site differences. If one
hypothesizes that the decomposers are very sensitive to S02 these differences
may have developed from the S02 fumigation. This hypothesis is supported by
the minimal difference noted between the S02 treatments by the 9 August sample
date. The significance of replication of the above investigation in 1976
cannot be overemphasized in light of the 1975 results.
364
-------
6-n
CD
CD
in
D
CD
c
o
o
w_
CD
Q-
30 r
20
Figure 10.11.
10
Control
Low
Medium
High
15
Apr
May
10
Jun
Jul
Aug
Sep
19
Decomposition of Agropyron smithii dead shoot material in nylon mesh litter bags for
Taylor Creek study sites, 1975.[Letter bags were placed in field 15 April and subsets
were retrieved on 10 June, 9 July, 8 August, and 19 September.)
-------
ARTHROPOD POPULATION AND BIOMASS DYNAMICS
Above- and belowground arthropods were collected on six dates in 1975.
Harvest and extraction techniques were the same as those described for the
Colstrip site monitoring program. The arthropod data have large statistical
errors resulting from sampling limitations. Since these data have not yet been
subjected to analyses of variance, the inferences made in the following
discussion are tentative.
Total arthropod numbers minus ants, which have been removed because of
erratic population estimates arising from clustered distribution, were
variable throughout the period sample (Figure 10.12). The LOW treatment had
the fewest number of arthropods and the CONTROL and HIGH treatments had the
largest numbers. No discernible trends related to fumigation are evident.
Examination of the species data indicated that thrips and mealybugs were
the most variable components. Figure 10.13 presents data without thrips or
mealybugs. Again the data were quite variable before fumigation, but post-
fumigation samples showed little variability among treatments and a general
decreasing trend for all treatments.
Trends in biomass were similar for the fumigated treatments with maximum
biomass occurring on the June sample data (Figure 10.14). In contrast to this,
peak biomass for the CONTROL treatment was observed in August. Because of the
large variability associated with arthropod biomass, the investigators are
hesitant to attribute this difference to treatment effects based on only one
year's data.
Table 10.6 shows a summary of all aboveground arthropods by feeding
strategy. Of the 251 species encountered, 50% were herbivores including tissue
and sap feeders, nectivores, and granviores; 25% were predators and parasites;
and 10% were scavengers or omnivores. The remaining species were either
nonfeeding pupae or species with unidentified feeding habits.
366
-------
500
Control
Low
Medium
High
April
Sept.
Figure 10.12.
Seasonal abundance of total aboveground arthropods for Taylor
Creek, 1975.
367
-------
500 r
400
300
O)
n
E
33
a 200
0)
100
Control
Low
Medium
High
c.
o
o
0>
e
3
0
April
May
June
July
Aug.
Sept.
Figure 10.13.
Seasonal abundance of total aboveground arthropods excluding
ants, Taylor Creek, 1975.
368
-------
CM
CP
£
CD
•o
o
CL
o
C/5
IT)
o
E
o
m
0
Control
Low
Medium
High
April
May
June
July
Aug.
Sept.
Figure 10.14.
Seasonal changes in aboveground arthropod biomass (excluding
ants), Taylor Creek, 1975.
369
-------
TABLE 10.6. TROPHIC DISTRIBUTION OF ARTHROPODS, TAYLOR CREEK, 1975,
Arthropods
Tissue feeders
Sap feeders
Predators
Scavengers
Parasites
Pollen and nectar
Omnivores
Seed feeders
Others
Total
No. of
Species
63
52
47
20
18
13
9
1
27
250
% of
Total
25.10
20.72
18.73
7.97
7.17
5.18
3.58
0.40
10.75
100.00
Belowground arthropods were divided into two size classes requiring
different sampling techniques. Macroarthropods included all soil insects
greater than 1 mm as well as spiders, centipedes, and millipedes, Microarth-
ropods included the mites and wingless insects less than 1 mm. The following
discussion is confined to the macroarthropod group. Analysis of the micro-
arthropod data is incomplete and does not merit discussion at this time.
Total macroarthropods based on time-weighted means for the four post-
fumigation sample dates indicated high variability of both biomass and numbers
among treatments. There were, however, a few groups that inhibited apparent
treatment response.
The Coleoptera (Figure 10.15) showed substantially reduced biomass and
numbers in the high treatment. Grasshopper eggs (Figure 10.15) were reduced in
numbers and biomass on all of the fumigated treatments, most likely the result
of avoidance by ovipositing females.
Predators (Figure 10.16) were the only trophic group to show a strong
treatment response and this was particularly evident in the predaceous
Carabidae (Figure 10.16) with a three-fold difference in both biomass and
numbers between the CONTROL and HIGH concentration treatment.
370
-------
100-
*l
E
x
Numbers
01
D 0
i i
P"1
n
•
•MB ',
c
Coleoptero (Total) 05On
"' O.5O-
N
E
«x^
3
0.25-
o
E
0
m
n-
^
PV
ff1
T
CO
30n
20 H
«n
I 10-
0-
Grasshopper Eggs
BCD
Treatments
N
I
O>
in
tn
o
E
o
0.02H
O.OH
A B C D
Treatments
Figure 10.15.
Numbers and biomass of selected soil macroarthropod groups (total Coleoptera and grass-
hopper eggs) for Taylor Creek, 1975. (A = CONTROL, B = LOW, C = MEDIUM, D = HIGH.)
-------
Prtdotors
r°-
Numbers
3_ 5
"
I
j
J
rri
:*_
— — »
CM
E
\
3
v>
to
a
E
o
£D
o.zoH
Q.iCH
A
B
CO
-^1
IN3
0)
v>
a
o
m
0.20 H
o.io-l
B
Figure 10.16.
w A B C D A
Treatments
Numbers and biomass of selected soil macroarthropod groups (predators and carabid
beetles) for Taylor Creek, 1975. (A = CONTROL, B = LOW, C = MEDIUM, D = HIGH)
-------
SUMMARY
In summary, continuous fumigation of a mixed prairie grassland during the
growing season has resulted in only subtle effects on primary producers and
arthropod consumers. Although the northern mixed-grass prairie may be
particularly resistant to sulfur dioxide fumigation, there are several reasons
why substantial changes were not recorded: the most important of these is that
fumigation was not begun until 1 June which was approximately 2 months after
the beginning of the growing season. These experiments are being continued
this year and fumigation was started on 10 April. These data will help explain
the 1975 results.
REFERENCES
Bell, J. N. B. and W. S. Slough. 1973. Depression of yield in ryegrass exposed
to sulphur dioxide. Nature. 241:47-49.
Brisley, H. R. and W. W. Jones. 1950. S02 fumigation of wheat with special
reference to its effects on yield. PI. Phys. 25:666-681.
Brisley, H. R. , C. R. Davis, and J. A. Booth. 1959. S02 fumigation of cotton
with special reference to its effect on yield. Agron. J. 51:77-80.
Daines, R. H. 1968. S02: Plant response. J. Occ. Med. 10(9):516-524. (Also:
1969 Air Quality Monographs, Monograph #69-8. 16 p. Am. Petrol. Inst. ,
NY).
Guderian, R. and H. Van Haut. 1968. Detection of S02 Effects upon plants.
Staub-Reinhalt. Luft. 30(1):22-35.
Guderian, R. and H. Schoenbeck. 1971. Recent results for recognition and
monitoring of air pollution with the aid of plants.
Hill, G. R. and M. D. Thomas. 1933. Influence of leaf destruction by sulfur
dioxide and clipping on yield of alfalfa. PI. Physiol. 8:223-245.
Hill, A. C. , S. Hill, C. Lamb, and T. W. Barrett. 1974. Sensitivity of native
desert vegetation to S02 and to S02 and N02 combined. APCA J. 24:153-
157.
National Research Council of Canada. 1939. Effect of sulfur dioxide on
vegetation. Ottowa, Canada. 447 p.
Thomas, M. D. and G. R. Hill. 1935. Absorption of S02 by alfalfa and its
relation to leaf injury. P. Physiol. 10:291-307.
Thomas, M. 1956. The invisible injury theory of plant damage. J. Air Pollut.
Contr. Assoc. 5:205-208.
Tingey, D. T. 1974. Ozone induced alterations in the metabolite pools and
enzyme activities of plants. In: Mack Dugger (ed.) Air Pollution Effects
on Plant Growth.
373
-------
CO
APPENDIX
PHENOLOGY OF MAJOR GRASS SPECIES, TAYLOR CREEK STUDY SITES, 1975
(A = CONTROL, B = LOW, C = MEDIUM, D = HIGH)
Date
27 May
3 Jun
10 Jun
1 6 Jun
22 Jun
30 Jun
7 Jul
13 Jul
21 Jul
30 Jul
4 Aug
11 Aug
18 Aug
26 Aug
2 Sep
10 Sep
15 Sep
Poa Stipa
secunda comata
ABCD ABCD
6666 5
8888 667
8888 7777
9988 111
9999 7777
9 10 9 10 9 9 9 10
11 11 11 11 10 10 10 10
13 13 13 13 12 13 12 12
14 14 13 14 13 13 14 13
14 13 14 14
14
Bromus
japonicus
A B
7 5
8 8
8 8
8 8
9 9
9 9
11 11
13 13
14 14
C
7
7
8
8
8
9
9
11
13
14
D
7
8
8
10
10
10
11
13
14
A
7
8
8
8
8
9
11
12
13
13
13
13
13
14
Koeleria
cristata
B C
7 7
8 8
8 8
8 8
9 9
9 9
11 11
12 12
13 13
13 13
13 13
13 13
13 13
14 14
Aristida
Bouteloua
longiseta
D
7
8
8
8
9
10
11
12
13
13
13
13
13
14
A
2
3
3
4
4
7
7
8
8
12
12
12
13
14
B
2
3
3
4
4
7
7
8
8
11
12
13
13
14
C
2
3
4
7
7
7
7
8
8
11
12
13
13
14
D
2
3
4
7
7
7
7
8
8
11
12
13
13
14
A
3
4
5
5
6
6
10
10
11
13
13
14
gracil is
B
4
4
5
5
6
6
9
10
10
11
13
13
14
C
4
4
5
5
6
6
9
10
10
11
13
13
14
D
4
4
5
5
6
6
9
10
10
11
13
13
14
Agropyron
srni thii
A
4
4
4
4
7
7
8
9
9
10
12
12
12
13
13
13
13
B
4
4
4
4
7
7
8
9
9
10
12
12
12
13
13
13
13
C D
4 4
4 4
4 4
4 4
7 7
7 7
8 8
9 9
9 9
10 10
12 12
12 12
12 12
13 13
13 13
13 13
13 13
(continued)
-------
APPENDIX (continued).
Date
27 May
3 Jun
10 Jun
1 6 Jun
22 Jun
30 Jun
7 Jul
£3 13 Jul
01 21 Jul
30 Jul
4 Aug
11 Aug
18 Aug
26 Aug
2 Sep
10 Sep
15 Sep
Taraxacum
off icinale
ABCD A
10 10 10 10 8
12 12 12 12 8
12 12 12 12 9
12121212 9
12 12 12 14 9
12 13 14 13
14 14 13
13
14
Antennaria
rosea
B
8
8
9
9
9
13
13
13
14
C
8
8
9
9
9
13
13
13
14
D
8
8
9
9
9
13
13
13
14
Tragopogon
dubius
A
5
7
7
8
8
10
12
12
12
13
14
B
5
7
8
g
9
10
12
12
12
13
14
C
5
7
7
9
9
10
12
12
13
13
14
D
5
7
8
9
9
10
12
12
13
14
Achillea
mi 1 1 ifol ium
A
9
9
10
10
10
10
11
13
14
B
g
9
10
10
10
10
11
13
14
C
9
9
10
10
10
10
11
13
14
D
9
10
10
10
10
11
13
14
Psoralea
Sphaeralcea
argophyl la
A
3
3
3
7
9
10
10
10
12
12
12
12
12
12
14
B
2
3
3
4
7
9
10
10
10
12
12
12
12
13
14
C
2
3
4
4
7
9
10
10
10
12
12
12
12
13
14
D
3
3
9
9
10
10
10
12
12
12
12
13
14
A
4
4
7
9
9
10
10
12
13
13
13
13
13
13
13
coccinea
B
4
4
4
9
9
10
10
12
13
13
13
13
13
13
13
C
7
7
7
9
9
10
10
12
13
13
13
13
13
13
13
D
4
7
7
9
9
10
10
12
13
13
13
13
A
5
5
5
6
6
6
7
7
8
8
9
9
10
Artemisia
fri
B
4
5
5
5
6
6
6
8
9
9
10
gida
C
4
5
5
5
6
6
6
6
7
8
8
9
9
10
D
5
5
5
5
6
6
6
6
7
8
8
8
9
10
-------
SECTION 11
MONITORING PLANT COMMUNITY CHANGES DUE TO S02 EXPOSURE
by
J. E. Taylor and W. C. Leininger
INTRODUCTION
This section describes how the plant community monitoring procedures
discussed in Section 2 have been used to monitor changes occurring on the Zonal
Air Pollution System (ZAPS) where known levels of sulfur dioxide (S02) exposure
have occurred.
DISCUSSION
Earlier studies (Taylor et aJL , 1975, 1976) have shown that Shannon-Weaver
and Simpson's indices worked best in the Col strip project situation. Other
indices (Redundancy, Probability of Inter- and Intra-Specific Encounters) were
used in 1975, but appeared to be insensitive or inconsistent in response to
conditions in this northern Great Plains ecosystem. The Shannon-Weaver indices
from ZAPS I were tested for statistical significance. Results are shown in
Table 11.1 and Figure 11.1.
The growing season conditions in 1975 were reflected in the diversity
indices. The spring was unusually cold, which delayed plant growth. This is
reflected in the relatively low indices observed in June and in the generally
lower values in 1975 compared with 1974. By July, diversity indices tended to
be higher, reflecting the abundant and diverse plant growth once growing
conditions became favorable. (There was good summer moisture). This was not
the case in the control plots. There was no significant difference among
Shannon-Weaver or Simpson's Indices for the control plots among sampling
periods. However, the indices associated with S02 application rates all
significantly increased in July and decreased in September. At the latter date,
all were significantly (P g .001) lower than those of the control.
At the July sampling period, the S02 plots showed diversity indices which
increased with their rates of gas application. This may be due to an
enhancement of plant growth in the period of early fumigation. By the September
date, however, the exact opposite was observed. All three S02 plots were
significantly (P ^ .001) less diverse than the control. Both indices showed
this effect. This appears consistent with the observations reported in the
remote sensing section (Section 7).
376
-------
0.9 i
X
LU
Q
CC
-LI
HI
0.8 -
0.7 -
0.6 H
0.5 -
0.51
-——- JUNE
~ - -JULY
— SEPTEMBER
0.70
0.56
HIGH
MEDIUM
LOW
CONTROL
Figure 11.1.
Shannon-Weaver index of diversity values for four S02
levels over three sampling periods, 1975.
377
-------
TABLE 11.1. STATISTICAL SIGNIFICANCE OF SHANNON-WEAVER INDICES1 AMONG ZAPS
PLOTS, 1975. (Values are averages of 2 lines of plots per
observation)
SAMPLING PERIODS
June
July
September
S02 Levels
CONTROL
LOW
MEDIUM
HIGH
0.6724
0.6059
0.6761
0.7689
MC
i I n fifinn
, ** , 0.6984
1 1
*
i i n 7.171
| | U . / 1 / \
***
i i n pcoq
| 1 U . Ou J o
NS
1 i
***
i i
I i
***
i i
1 I
***
i i
I i
0.6972
0.5560
0.5856
0.5116
Significance of t-test comparing values connected by line.
NS = no significant difference
* = P ^ .05
** = p g ,oi
*** = p g .001
1 = Calculated with Log base 10.
The underlying causes of the diversity differences still are unclear be-
cause of the confounding influences of pre-treatment plant community pattern and
density, coupled with treatment-induced changes in both species and numbers of
individuals.
Diversity indices for the ZAPS sites are shown in Figures 11.2 and 11.3 for
1975 and 1976, respectively.
In 1975, the growing season was delayed by late storms and cold tempera-
tures. The cool-season species were not yet at peak development at the June
sampling date. This could explain the higher diversities observed in July at
this location. (This was not seen at Colstrip, where elevations and precipi-
tation are lower). The September decrease in diversity follows the usual pat-
tern. There are no obvious diversity differences among S02 levels, with the
possible exception of an enhancement on the high rate in July. If this is a
real difference, it may reflect a transitory enhancement of plant growth with
low levels of exposure to S02.
A more typical growing season occurred in 1976.
in diversity followed the expected trend.
Thus, the seasonal changes
It is obvious that the ZAPS II site is inherently less diverse than ZAPS I.
It has lower canopy cover values and fewer species. This is illustrated in the
diversity values between the ZAPS locations (Figure 11.3). Also, ZAPS I re-
ceived more precipitation than ZAPS II in 1976, which could contribute to the
difference.
378
-------
3.0
2.5 -
2.0 -
1.5 -
JUNE
JULY
SEPTEMBER
HIGH
MEDIUM
LOW
CONTROL
LEVELS OF SO2
Figure 11.2. Shannon-Weaver function (HP) for ZAPS I, three sampling
dates, 1975.
3-0 -i ZAPS I
ZAPS n
2.5 -
2.0 -
1.5
HIGH
LOW
CONTROL
MEDIUM
LEVELS OF SO2
Figure 11.3. Shannon-Weaver function (HP) for ZAPS I and II, two
sampling dates each, 1976.
379
-------
The depression of diversity observed on the low S02 plots on ZAPS II is
similar to the response of ZAPS I during its first year of fumigation (1975).
Whether the second year of ZAPS II will show a corresponding recovery remains to
be seen.
ZAPS SITE SIMILARITY
Canopy coverage data (means of 40 frames per plot) were used to calculate
Similarity Coefficients among the ZAPS plots, using the procedure of Sokal and
Sneath (1963). This was done for June and August, 1976. Similarity dendrograms
are presented in Figures 11.4 and 11.5.
In June, there was a clear separation of the two ZAPS sites. Also, in both
cases the vegetation on the CONTROL and LOW treatment plots was similar (0.89
and 0.99), as was that on the MEDIUM and HIGH S02 level plots (0.90 and 0.89).
Overall similarity within sites was 0.78 in ZAPS I and 0.71 in ZAPS II.
In August, the same trends are evident. Both CONTROL and LOW plots
remained similar. The MEDIUM and HIGH were similar on ZAPS I, and somewhat less
so on ZAPS II. The sites were considerably more similar in August than in June,
probably due to the lower overall species number observed at the later date.
PHENOLOGY STUDIES
Phenological stages occurring on the ZAPS plots are shown graphically in
Figures 11.6 and 11.7.
In 1975 no consistent phenological differences were observed among ZAPS
plots for any species. This may be an artifact of sampling, since the aerial
monitoring did show a definite tonal gradation across the plots as the season
progressed. This inconsistency may be due to inadequate sampling frequency,
relative insensitivity of field procedures, or the avoidance of damaged plants
in the immediate vicinity of the gas delivery pipes in an attempt to minimize
"bias" in the sample. The investigators feel that there may in fact have been no
important phenological differences, but that the reflected differences were due
to bleaching of the stressed vegetation as it matured.
SUMMARY
The Shannon-Weaver Index based on both number and on cover *s sensitive to
different S02 levels and seasonal plant composition. The early effect of S02
fumigation appears to be an enhancement of plant species diversity. By late
season, 1975, this tendency was reversed, and the diversity based on numbers for
all three S02 levels were significantly (P ^ .001) less diverse than the
control.
Cover-based diversity shows similar trends for seasons and sites, and will
be utilized further in future work.
Species similarity coefficients show a tendency to group the CONTROL and
LOW versus the MEDIUM and HIGH ZAPS treatments on both sites and both dates.
380
-------
1.0
.9
UJ .7
O
O
5.5
CO .4
0
i i i IJIJIUE
CONTROL LOW MEDIUM HIGH CONTROL LOW MEDIUM HIGH
12345678
L .9869 |
.89O4
.7762
.8933
.7140
.4654
Figure 11.4. Dendrogram of similarity coefficients for the ZAPS sites,
based on canopy coverage, August 1976.
1.0
2.9
I-
•z.
y s
O
£.7
O
O
> .6
E
< .5
CO -4
0
i i n n n n i i
CONTROL LOW HIGH CONTROL LOW MEDIUM MEDIUM HIGH
2345678
.8169
7924
.9IO4
.8256
.7851
.8577
.7819
Figure 11.5.
Dendrogram of similarity coefficients for the ZAPS sites,
based on canopy coverage, June 1976.
381
-------
DEAD
WINTER DORMANCY
FALL GREENUP
MATURITY
SEED SHATTER
FRUIT FORMED
LATE FLOWERING
FLOWERING
FLOWER BUDS OPENING
SHOOTING SEED STALK
FLOWER BUDS APPEARING
BOOT STAGE
VEGETATIVE GROWTH
EARLY GREENUP
BASAL ROSETTE
SEEDLING
COTYLEDON
Agropyron smithii
Aristida longiseta
Koelfria ens fata
Poa sandbergii
Stipa comata
ANNUAL BROMES
MAY
15
JUNE JULY
34 12
AUG.
28
NOV.
3
Figure 11.6. Phenological profile of selected species on Taylor Creek
control plot.
382
-------
DEAD
WINTER DORMANCY
FALL GREENUP
MATURITY
SEED SHATTER
FRUIT FORMED
LATE FLOWERING
FLOWERING
FLOWER BUDS OPENING
SHOOTING SEED STALK
FLOWER BUDS APPEARING
BOOT STAGE
VEGETATIVE GROWTH
EARLY GREENUP
BASAL ROSETTE
SEEDLING
COTYLEDON
,„.—_ Artemisia frigida
— Plantago patogonica
^—— Sphaeralcea coccinea
Taraxacum officinale
Tragopogon dubius
(NEW GROWTH,TRDU)
MAY
JUNE JULY
24 12
AUG.
28
NOV.
3
Figure 11.7. Phenological profile of selected species on Taylor Creek
control plot.
383
-------
The phenology scoring system has proved very satisfactory in a variety of
vegetation types. However, phenology changes attributable to SCL have not been
quantified because of the many confounding influences. The frequency and
intensity of sampling is being increased to construct a more complete data base
so that confounding influences can be identified and separated.
REFERENCES
Sokal, R. R. and P. H. A. Sneath. 1963. Principles of numerical taxonomy. W.
H. Freeman, San Francisco.
Taylor, J. E. , W. C. Leininger, and R. J. Fuchs. 1975. Baseline vegetational
studies near Colstrip, Proc. Ft. Union Coal Field Symp. , Mont. Acad. Sci.
pp. 537-551.
Taylor, J. E. , W. C. Leininger, and R. J. Fuchs. 1976. Monitoring plant
community changes due to emissions from fossil fuel power plants in eastern
Montana. Section II of the bioenvironmental impact of a coal-fired power
plant, second interim report. USEPA Ecological Research Series EPA-600/3-
76-013. pp. 14-40.
384
-------
SECTION 12
EFFECTS OF LOW-LEVEL S02 STRESS
ON TWO LICHEN SPECIES
by
S. Eversman
INTRODUCTION
As part of the development of western energy resources, coal-fired power
plants are proliferating in the Great Plains region. Two 350-megawatt
generating plants began operation in autumn, 1975, and spring, 1976, in
Colstrip in southeast Montana. Two more are in the planning stages.
Sulfur dioxide (S02), one of the emissions from burning coal, has been
repeatedly demonstrated to cause adverse effects in plants, especially
lichens. Many laboratory experiments have shown chlorophyll bleaching,
decreases in respiration and photosynthetic rates, and plasmolysis with S02
exposure (LeBlanc and Rao, 1975). The lowest, experimental S02 concentration
has been at 0.50 ppm for 12 hours (Nash, 1975).
LeBlanc and Rao (1975) cite experimental works that draw these
conclusions. Transplanting lichen species into areas already polluted results
in lichens exhibiting the same internal and external pathological symptoms
demonstrated in laboratory tests. Most of the studies have been in humid areas
where S02 is thought to be more harmful than in semi-arid climates, They
estimate that, in humid climates, long-range average concentrations of S02
below 0.002 ppm would cause no damage to lichen species; acute damage occurs
above 0.03 ppm, and intermediate amounts of S02 would cause chronic damage to
lichens.
This report presents results of observations on the effects of chronic
low-level S02 stress on two lichens native to semi-arid grassland and ponderosa
pine vegetation types. The objectives of the study were to:
1) Identify and time the anatomical and physiological changes that
occurred with exposure to various low levels of S02, and
2) to make simultaneous observations on associated vascular
vegetation for comparison purposes.
Usnea hi rta is an epiphyte on trunks and branches of ponderosa pine, the
major tree of this region It is a fruticose lichen, i.e., stringy in
appearance; this type of lichen is considered particularly susceptible and
sensitive to air pollutants.
385
-------
Parmelia chlorochroa is a foliose (leafy, flat) lichen that lives on bare
soil between grass clumps and shrubs on the grasslands. Foliose lichens are
also considered sensitive to air pollution.
A fumigation system was constructed on a grassland site in southeast
Montana. The system was composed of 4 plots, each about 0.654 hectare. Each
plot had a network of aluminum pipes that delivered different amounts of S02
through small holes in the bottoms of the pipes. Lee and Lewis (1976) fully
describe the system. One system of four plots was in operation in 1975 and
1976, and a duplicate system operated in 1976 about 1 km from the first site.
This report refers to the system operating in both 1975 and 1976 as ZAPS I (I =
Site 1, ZAPS = Zonal Air Pollution System). The fumigation or treatment plots,
within each site or system, are referred to as A (CONTROL), B (LOW), C (MEDIUM)
and D (HIGH) (Lee and Lewis, 1976). ZAPS II refers to the second system, with
similar treatment plots A, B, C and D, operating in 1976.
By specifically identifying the characteristics of the two lichen species
exposed to the S02 stress on the ZAPS sites, these species can be used as
biological systems to monitor increasing S02 content of the air in the Colstrip
area. Simultaneous observations made by other researchers in the region will
be combined with the lichen observations to identify and interrelate ecosystem
components affected by decreasing air quality.
Usnea hirta and Parmelia chlorochroa from several ponderosa pine and
grassland field study sites have been collected and observed for baseline
conditions. They have been compared with the stressed lichens from the ZAPS
sites. The sites are mapped and described in a previous chapter (Eversman,
1976).
METHODS
Five metal fenceposts were placed nonrandomly in each of the ZAPS
fumigation plots. They were placed toward each corner and in the center of
each plot where there was minimum grass cover so that Parmelia chlorochroa
could be transplanted to the soil. There was a minimum of 2 meters between a
fencepost and any fumigation pipe.
Transplants of both Usnea hirta and Parmelia chlorochroa were made 1 May
1975 and 23 March 1976; fumigations began 24 May 1975 and 1£ April 1976.
Ponderosa pine branches containing U. hirta were wired onto the northeast side
of each fencepost so the branches extended about 0.25 - 1.0 meters vertically
on the post. P. chlorochroa was moved from the soil in a nearby field (Site P7)
to the fumigation sites to augment the existing populations. A second set of
P. chlorochroa transplants was made 17 July 1976 from Site G5 (Kluver East) to
compare vertical position effects of the S02 stress. P_. chlorochroa samples
were tied on bare ponderosa pine branches and wired to the fenceposts on Site
1, parallel to the branches containing U. hirta. Duplicate samples were placed
on the soil at the base of each fencepost.
Samples were collected about once a month through September of each year.
They were observed for general thallus condition and photographed. Respiration
rates were determined manometrically (Eversman, 1975). Samples were sent to
386
-------
the Montana State University Soil Testing Laboratory for sulfur analysis. The
details of analytical methods are presented in the Appendix.
Plasmolysis counts were made by preparing 3 wet-mount slides of 3
different plants and counting 100 algal cells on each slide, recording the
number of plasmolyzed cells.
The same set of observations was made on lichen samples collected at about
the same times, 20-97 km from the fumigation sites.
Stems and leaves of Agropyron smithi i and Koeleria cristata, two grasses,
were collected near each fencepost on Sites 1 and 2 for 3 months in 1976, to
determine respiration rates and sulfur content.
RESULTS
Respiration rates and results of the completed plasmolysis counts for
Usnea hirta appear in Table 12.1. Samples from the A (CONTROL) plot on both
Sites 1 and 2 did not exhibit respiration rate and plasmolysis percentages
different from those of U. hirta samples collected simultaneously from their
natural habitats.
The respiration rates of samples from Plot B were consistently below those
of samples from the control site, but within the range of samples from all
unpolluted sites. The plasmolysis level reached about 76% of the control
levels in 100 days, and leveled off throughout the 156 days of S02 treatment.
Plasmolyzed algal cells are also bleached.
The respiration rates of U. hi rta samples from plot C decreased to 57% of
those of control plot A samples, and remained at less than 50% of control rates
for the remainder of the test period. Plasmolysis was nearly 100% in 31-33
days, and the algae never recovered.
Samples from plot D exhibited a respiration rate that was 6% of that of
control samples in 1975, but in 1976 the rates fell to 33-46% of control plot A
samples. They remained 19-36% of those of control samples for the test period.
Plasmolysis and bleaching reached 100% in 31-33 days, with no recovery.
P_. chlorochroa samples on the ground did not exhibit changes in
respiration rates in any of the treatment plots (Table 12.2). The results of
plasmolysis counts are still very incomplete, but some representative counts
showed a slight increase of plasmolysis with S02 stress. When samples were
elevated to the fenceposts with the L). hirta, however, plasmolysis increased to
96% in 23 days on plot D, and the mean respiration rate dropped to 22% of that
of control (A) samples. Respiration rates of elevated P. chlorochroa decreased
on plots B, C, and D; the rates of control samples, A, and those on the ground
on all plots, did not decrease.
Grass samples exhibited a generally decreasing respiration rate on all
plots from June to August (Table 12.3). This decrease does not superficially
appear to be related to S02 dosage.
387
-------
TABLE 12.1. RESPIRATION RATES AND PERCENTAGE OF PLASMOLYSIS FOR Usnea hirta
ON ZAPS SITES AND ON SOME COLSTRIP FIELD SITES, 1975-76. Respiration rates
are given as the mean of 3-10 samples ± 1 standard deviation, expressed in ul
02 consumed/g dry weight/hour. Plasmolysis is expressed as mean percentage of
3 samples ± 1 standard deviation.
Collection Site
P10, East Otter Creek
P9, transplant source
PI 5, Fort Howes natives
PI 5, Fort Howes natives
P16, Poker Jim Butte
ZAPS: IA
IIA
IB
IIB
1C
IIC
ID
IID
P6, Kluver West tr.
ZAPS: IA
IB
1C
ID
P16, Poker Jim Butte
P13, Home Creek Butte
Pll, SEAM 1
ZAPS: IA
IB
1C
ID
P10, East Otter Creek
ZAPS: IA
IIA
IB
IIB
1C
IIC
ID
IID
P9, Road
Number of
S02 Treatment
Days, Date
0
0
0
0
0
31
M
ii
H
M
M
n
ii
0
33
M
n
n
0
M
M
47
n
M
n
0
72
n
n
n
n
M
M
II
0
3-23-76
5-01-75
M
5-13-76
n
n
n
n
n
n
n
n
n
6-25-75
n
n
M
n
7-16-75
n
n
7-10-75
n
M
n
6-23-76
n
n
n
n
n
n
n
n
8-11-75
Resp.
Rate
X
709
563
511
576
694
711
800
718
755
290
438
325
264
864
821
746
472
53
630
462
545
686
555
333
0
744
704
555
359
476
236
263
115
169
508
1
s.d.
63
109
213
113
183
72
97
141
74
155
175
199
99
86
37
98
204
45
77
55
35
115
121
307
0
45
71
34
78
77
83
48
67
38
50
Plasm.
% x 1 s.d.
3
6
4
35
37
94
99
94
100
6
54
99
100
11
4
4
63
58
100
99
100
100
3
1
3
25
39
9
2
9
0
4
20
1
0
2
2
2
18
21
1
1
1
1
(continued)
388
-------
TABLE 12.1 (continued)
Number of
S02 Treatment
Collection Site Days, Date
ZAPS
P10,
P16,
Pll,
ZAPS
P10,
P8,
P3,
ZAPS
P10,
P14,
P14,
ZAPS
P10,
P15,
P15,
ZAPS
: IA
IB
1C
ID
East Otter Creek
Poker Jim Butte
SEAM 1
: IA
IIA
IB
IIB
1C
IIC
ID
IID
East Otter Creek
Morning Star View
Kluver NE1
: IA
IB
1C
ID
East Otter Creek
3-Mile top
3-Mile bottom
: IA
IIA
IB
IIB
1C
IIC
ID
IID
East Otter Creek
Fort Howes transplants
Fort Howes natives
: IA
IIA
IB
IIB
1C
IIC
ID
IID
79
11
n
ii
0
n
n
96
ii
ii
ii
ii
M
ii
n
0
n
n
110
II
II
II
0
II
II
119
n
n
M
n
n
M
n
0
II
II
156
n
M
M
n
n
n
M
n
n
M
n
7-15-76
II
II
7-17-76
n
n
M
n
n
n
n
9-25-75
M
M
9-11-75
n
n
M
8-09-75
n
M
n
n
n
n
n
n
n
n
9-15-76
n
n
9-14-76
n
n
n
n
M
n
n
Resp.
Rate
X
738
435
316
0
463
610
453
670
643
473
479
323
264
244
98
640
828
736
709
499
355
0
668
667
547
721
618
556
509
298
282
150
118
633
597
470
631
506
487
457
340
289
142
112
1
s.d.
66
87
133
0
57
31
19
103
38
39
86
35
49
92
20
40
135
72
55
128
137
0
41
76
40
18
50
84
66
73
84
29
16
53
43
50
21
61
43
37
40
85
68
46
Plasm.
% x 1 s.d.
9
5
5
69
66
99
99
100
100
15
77
98
100
5
24
10
20
4
82
71
100
99
100
100
3
2
2
17
18
2
2
0
0
3
16
3
0
2
10
1
5
2
13
18
1
1
0
0
389
-------
TABLE 12.2. RESPIRATION RATES AND PERCENTAGE OF PLASMOLYSIS FOR Parmelia
chlorochroa ON SOME COLSTRIP FIELD SITES, 1975-1976. Respiration rates are
given as the mean of 3-10 samples ± 1 standard deviation, expressed in yl 02
consumed/g dry weight/hour. Plasmolysis is expressed as mean percentage of
3 samples ± 1 s.d.
Collection Site
Field near ZAPS sites
G7, Pasture
Gl , Hay Coulee
G7, Pasture
ZAPS: IB
1C
G5, Kluver East
Gl , Hay Coulee
ZAPS: IA
IB
1C
ID
G4, Kluver North
G3, Kluver West
ZAPS: IA
IB
1C
ID
Field near ZAPS site
ZAPS: IA
IB
1C
ID
Gl , Hay Coulee
G3, Kluver West
G4, Kluver North
G5, Kluver East
ZAPS: IA (native)
IIA (native)
IB
IIB
IIC
IID
Number of
S02 Treatment
Days , Date
0
"
0
11
31
I!
0
n
33
II
1!
II
0
II
47
II
II
II
0
79
II
II
ii
0
n
n
n
96
"
M
11
n
M
5-01-75
II
5-14-76
n
"
"
6-26-75
11
n
11
11
11
7-07-75
n
7-10-75
II
II
II
8-11-75
II
II
II
M
7-14-76
n
n
M
7-17-76
II
"
M
II
II
Resp.
Rate
X
223
337
458
371
276
290
316
215
319
330
345
296
349
293
345
404
317
294
217
356
357
336
309
299
272
242
322
231
293
299
262
301
335
1
s.d.
39
132
43
54
50
15
40
26
44
36
36
54
21
29
80
54
50
38
28
48
36
47
37
22
25
4
26
41
36
22
31
30
22
Plasm.
% x 1 s.d.
2 2
8 2
10 7
4 4
43 34
22 3
(continued)
390
-------
TABLE 12.2. (continued)
ZAPS
ZAPS
Collection Site
: IA
IB
1C
ID
: IA (native)
IID
After new transplants
ZAPS
ZAPS
ZAPS
ZAPS
Gl,
G3,
G4,
G5,
ZAPS
ZAPS
ZAPS
ZAPS
: IA
IIA
IA*
: IB
IIB
IB*
: 1C
IIC
1C*.
: ID
IID
ID*
Hay Coulee
Kluver West
Kluver North
Kluver East
: IA (native)
IA
IIA
I A*
: IB
IIB
IB*
: 1C
IIC
1C*
: ID
IID
ID*
Number of Resp.
S02 Treatments Rate
Days , Date x
110 9-11-75
n n
i n
M n
119 8-09-76
n M
were made 7-17-76 (* =
23 8-09-76
n n
n n
n n
n n
n n
n n
n n
n n
ii
M M
M n
0 9-14-76
n n
n n
n n
60 9-15-76
n n
n n
n n
N n
n n
n n
n n
M II
II II
II II
II II
II II
362
271
353
340
350
293
elevated
298
297
366
314
315
311
309
368
197
396
437
79
274
301
262
239
315
305
326
345
270
338
174
322
403
45
396
386
43
1 Plasm.
s.d. % x 1 s.d.
69
20
52
21
20
50
samples):
8
28
26
25
40
33
26
30
54
24 156
38
15 96 3
22
15
34
15
31
44
31
9
33
28
47
38
31
10
13
77
34
391
-------
TABLE 12.3
RESPIRATION RATES
OF TWO GRASSES, Koeleria cristata AND Agropyron
smithi i, FROM THE ZAPS
SITES, 1976. Respiration rates are given
as the mean of 3 samples ± 1 standard deviation, expressed in yl
02 consumed/g dry weight/hour.
Koeleria
cristata
Col lection
Plot
IA
IIA
IB
IIB
1C
IIC
ID
IID
IA
IIA
IB
IIB
1C
IIC
ID
IID
IA
IIA
IB
IIB
1C
IIC
ID
IID
Number of
S02 Treatment
Days, Date
72 6-23-76
n M
n M
n n
n n
n n
M n
n n
96 7-17-76
n n
n M
n n
n M
n M
n n
M n
119 8-09-76
n n
M n
M
n M
M n
n n
n N
Resp.
Rate
X
702
706
706
617
893
564
1045
759
506
247
587
411
560
430
827
525
128
147
164
226
146
375
108
332
1
s.d.
51
103
102
134
104
120
42
114
4
8
130
75
72
28
113
103
35
25
19
45
29
41
31
46
Agropyron
smithi i
Resp.
Rate
X
646
579
736
699
516
551
610
349
355
335
231
328
197
366
212
249
145
312
177
107
239
394
297
530
1
s.d.
189
30
163
70
106
34
235
34
64
38
90
120
64
163
no
55
13
38
16
32
29
50
68
85
392
-------
The data from the S04 content analysis from 1975 (Table 12.4) suggest
increasing sulfur content as S02 exposure continued, but available results are
inconclusive. The 1976 analyses are not yet complete.
Microscopic examination revealed extensive bacteria populations
associated with the lichens, especially with ?_. chlorochroa. As 562 stress
increased in dosage and time, the bacteria appeared to be increasingly attached
to the fungal hyphae, rather than in the water medium (on a microscope slide).
The bacteria were associated less with algae than with the fungal hyphae.
DISCUSSION
A reasonable question arises in connection with both the Usnea hirta and
Parmelia chlorochroa transplants on the control (A) plots.TFe IIflirta
specimens have been transplanted from ponderosa ,pine sites that are shadier
than the fumigation sites and control plot A. However, U. hirta is not a plant
of deep shade; it grows on partially exposed branches and trunks of ponderosa
pine. Every effort was made to compare the measurements made on U. hirta from
control plots A with those of native samples collected directly from their
native habitats. The respiration rates, plasmolysis counts, and visual
appearance of the U. hirta exhibit no harmful effects from the transplanting.
P. chlorochroa is a native of exposed bare soil and was transplanted in
most cases to_a similar habitat. Some adverse effects might have been expected
when it was tied to pine branches and elevated above the soil, but this was not
the case. A common observation when any specimens are transplanted is a slight
temporary increase in respiration rate over that of natives or duplicate
samples, but the increase is not permanent and no visual effects can be
detected.
Decreasing respiration rates, algal bleaching and plasmolysis, and
yellowing of the plant U. hirta reflect increasing S02 exposure.
The respj ration rates of P. chlorochroa samples on the ground did not
decrease with the S02 treatments"nor did the plants change color. P.
chlorochroa specimens were then placed on the branches 0.75 m above the ground
to test if: 1) vertical position in the grassland stratification had any
effects and 2) associated bacteria decreased when the thalli were off the
ground. The bacterial populations remained visually the same but the
respiration rates decreased dramatically in proportion to S02 dosage, and the
percentage of plasmolysis increased to nearly 97% in 23 days on plot D.
This seems to indicate that the S02 levels on the sites vary appreciably
from ground level to 0.5-1.0 m above the ground, although compounding stress
effects could be a factor in the observed reactions. Further work must include
using some monitoring method, possibly sulfation plates, to detect the vertical
distribution of the S02 on the treatment plots.
Conclusions reached before £. chlorochroa samples were placed on the
branches on the test site fenceposts were that U. hirta is a species more
sensitive than P. chlorochroa to S02 stress. Observations of elevated P.
chlorochroa sampTes indicate that both species are sensitive to S02- However,
393
-------
TABLE 12.4. SULFATE SULFUR CONTENTS OF Usnea hirta AND Parmelia chlorochroa
SAMPLES COLLECTED IN 1975. Expressed as mean percentage ± 1
standard deviation of 3 - 5 samples. Determined by the MSU
Soils Testing Laboratory.
Usnea hirta
Number of
S02 Treatment %
Collection Site
P9, Road
P15, Fort Howes tr.
PI 5, Fort Howes natives
P9, Road
P3, Kluver NE1
P8, Morning Star View
P13, Home Creek Butte
P10, East Otter Creek
ii n
ZAPS: IA
IB
IB
IB
1C
1C
1C
ID
ID
Parmelia chlorochroa
Pasture near ZAPS sites
G7, transplant source
ZAPS: IA
IB
IB
IB
1C
1C
ID
ID
Days
0
n
n
0
0
0
0
II
0
47
47
79
110
47
79
no
47
79
0
n
47
47
79
110
47
no
47
79
, Date
5-01-75
n
ii
8-11-75
9-25-75
M
7-16-75
M
9-25-75
7-10-75
7-10-75
8-11-75
9-11-75
7-10-75
8-11-75
9-11-75
7-10-75
8-11-75
5-01-75
M
7-10-75
7-10-75
8-11-75
9-11-75
7-10-75
9-11-75
7-10-75
8-11-75
Sul fur
0.25
0.25
0.19
0.26
0.03
0.19
0.15
0.08
0.22
0.29
0.30
0.17
0.29
0.33
0.19
0.37
0.32
0.41
0.17
0.19
0.23
0.19
0.18
0.20
0.23
0.18
0.21
0.22
1 s.d.
0.04
0.03
0.04
0.04
0.01
0.08
0.16
0.01
0.11
0.01
0.03
0.15
0.11
0.03
0.11
0.07
0.05
0.01
0.02
0.03
0.04
0.02
0.02
0.03
0.03
0.03
0.03
0.01
394
-------
U. hirta may be more useful as an area-wide indicator of decreasing air quality
because of its more exposed locations on tree trunks and branches. Wind
conditions or filtering by grasses and forbs may prevent a great deal of $62
from reaching ground level, except in bare areas, where it can demonstrably
affect P. chlorochroa.
Simultaneous visual observations were made on the associated vascular
vegetation on the fumigation plots. While unmistakable bleaching was evident
in U. hirta on all test plots (ZAPS I and II, B, C, and D plots) 31-33 days
after the fumigation began, no visual effects appeared on the grasses during
the test periods. The height of the grass plants was about equal to the
vertical placement of the transplanted lichens. The generally decreasing
respiration rates of Agropyron smithii and Koeleria cristata are probably a
function of their phenology.The rate of decrease is similar on nearly all
plots, both sites, from June to August. Exceptions (samples from 2D and 1C,
August, 1976), however, might reflect S02 stress, which has been demonstrated
to raise respiration rates when present in very small amounts (LeBlanc and Rao,
1975).
Other personal observations were leaf-tip curling and browning in
Achillea lanulosa after 60 days of stress on plot ID in 1975. The grasses and
forbs oh stress plots B, C, and D, Site 1, cured a much yellower color
overwinter (1975-1976) than did adjacent non-stressed vegetation.
395
-------
REFERENCES
Eversman, Sharon. Lichens as Predictors and Indicators of Air Pollution from
Coal-Fired Power Plant Emissions. The Bioenvironmental Impact of a Coal-
Fired Power Plant, Second Interim Report, Colstrip, Montana. Corvallis
Environmental Research Laboratory, Office of Research and Development,
U.S. Enviromental Protection Agency, Corvallis, 1975.
Eversman, Sharon. Soil and Epiphytic Lichen Communities of the Colstrip,
Montana Area. The Bioenvironmental Impact of a Coal-Fired Power Plant,
Third Interim Report, Colstrip, Montana. Corvallis Environmental
Research Laboratory, Office of Research and Development, U.S.
Environmental Protection Agency, Corvallis, 1975.
LeBlanc, Fabius and D.N. Rao. Effects of Air Pollutants on Lichens and
Bryophytes. Iji Mudd, J.B. and T.T. Kozlowski (Eds). Responses of Plants
to Air Pollution, Academic Press, New York, 1975.
Lee, Jeffrey, J. and R.A. Lewis. An Experimental System for Testing the
Effects of Sulfur Dioxide on Grassland Systems. In press. 1976.
Nash, Thomas H. Influence of Effluents from a Zinc Factory on Lichens.
Ecological Monographs 45 (2): 183-198, 1975.
Stoltenberg, Larry E. Availability of Sulfur to Alfalfa (Medicago sativa L.)
and Orchardgrass (Dactyl is glomerata L.) as Influenced by Source and Time
of Sulfur Fertilizer Applications. Unpublished MS Thesis. Plant and
Soils Department, Montana State University, 1969.
396
-------
APPENDIX
The method used by the Montana State University Soils Testing Laboratory
for analysis of plant sulfur content is outlined below. The method is
described in Stoltenberg (1969).
This procedure is a modification of one that originally appeared in:
Bardsley, D.E. and J.D. Lancaster. Determination of Reserve Sulfur and Soluble
Sulfates in Soils. Soil Science Society of America Proceedings 24: 265-268.
July-August, 1960.
1. Grind dry plant material in a Wiley mill, with 20-mesh screen.
2. Mix 0.250 g plant + 1 g ashing powder in 50 ml beaker.
3. Add 0.5 g ashing powder as topdressing to minimize loss of volatiles.
a. Ashing powder: 27 g Na2C03
3 g NaHC03
1.8 g NaN03
0.06 g KMn04
b. These are ground in power mortar and pestle until no greater color
intensity is observed from KMn04 (about 5 minutes).
4. Beakers are set in a muffle furnace for 1 hour at 260°C, then 2 hours at
480°C.
5. Ash is treated with 25 ml acid solution.
Acid solution: a. 114.2 ml cone HC1
125.6 ml cone CH3COOH
8.3 g NaH2P04 -H20
b. Make to 2 liters with distilled water.
6. Ash and acid are shaken for 30 minutes.
7. 5 drops of H202 are added to each flask for clearing.
8. Mixture is filtered through Whatman #40 double acid washed filter paper.
9. 10 ml filtrate is pipetted into 125 ml Erlenmeyer flask, and 1 ml acid
seed solution is added.
397
-------
Acid seed solution: 0.5434 g K2S04 in 1 liter 6N HC1 until 20 ppm S.
10. 0.5 g BaCl2 is added to filtrate.
11. Flasks rest for 1 minute, then swirled until BaCl2 crystals dissolve.
12. Solution is read in Bausch & Lomb colorimeter within 2-8 min. at 420 nm.
398
-------
SECTION 13
EFFECTS OF LOW-LEVEL S02 EXPOSURE ON SULFUR ACCUMULATION AND VARIOUS PLANT
LIFE RESPONSES OF SOME MAJOR GRASSLAND SPECIES UNDER FIELD CONDITIONS
by
C. C. Gordon, P M. Rice, and P. C. Tourangeau
INTRODUCTION
During the 1975 growing season, S02 fumigations of a cool season-short
grass ecosystem were initiated on ZAPS I (Zonal Air Pollution System) from May
28 to September 28. Because of the visible changes across the four plots (0, 2,
5, and 10 pphm ) on ZAPS I in October of 1975 which were not accompanied by any
significantly measurable changes in biomass or species diversity (Dodd et al.,
1976), a substantial portion of the previous efforts and studies on the
ponderosa pine-skunkbush sites were redirected to the ZAPS studies during 1976.
The original 1975 hypothesis for the ZAPS fumigation studies was that most plant
species fumigated continuously with MEDIUM and HIGH S02 concentrations on these
plots during the growing season would be severely damaged or killed outright.
This hypothesis was based on the S02-damaged forests studied by Linzon (1971,
1973a) and Dreisinger (1965, 1970) in the vicinity of Sudbury and Biersdorf,
Ontario, Canada, where the highest S02 annual concentration was .044 ppm (4.4
pphm). The investigators also considered studies done in Anaconda, Montana,
where a large stationary source of S02 emissions is located and where severe air
pollution damage to vegetation occurs each year; those sites which record the
highest S02 readings average only .02 ppm or less annually. Thus the hypothesis
that the cool season-short grass ecosystem on the ZAPS plots would be severely
damaged or killed outright on the higher S02 delivery plots was based on air
pollution studies in these other two polluted areas.
Thus far the 1975 hypothesis has not proved to be valid, so a new hypoth-
esis was formulated before the 1976 growing season began to test in the 1976
field studies on the ZAPS sites. This new hypothesis was that the plant species
of cool season-short grass ecosystems which are fumigated continuously with S02
during the growing season will manifest early senescence, loss of viable seed
production, and excessive quantities of sulfur throughout all plant parts
(roots, stems and leaves).
To test this hypothesis, the investigators established several study
objectives which encompass the following: (1) Bimonthly collections of five to
six species of plant foliage during the growing season; (2) collection of roots
of two grass species (one rhizomatous and one bunchgrass species) at mid and
late growing season periods; (3) analysis of all plant parts for sulfur concen-
399
-------
trations; (4) data gathering on leaf emergence, growth, and death of western
wheatgrass (Agropyron smithii) on all plots at both the ZAPS I and II sites
during the growing season to ascertain any difference in phenology by S02
concentrations, and (5) collection of mature seeds from various species of
grasses, fumigated with the different S02 concentrations, at the end of the
growing season for seed germination tests ,under laboratory conditions.
MATERIALS AND METHODS
1975 VEGETATION COLLECTIONS
ZAPS I first was sampled on June 28. On September 28, both ZAPS I and the
area where ZAPS II was to be built were sampled. Each of the four treatment
plots on ZAPS I was subdivided into six subplots. A composite grab sample of
each species from each treatment was made by collecting material from each
subplot. The June sample was composed predominantly of live vegetation while
the September sample consisted of cured vegetation, except the Gutierrezia
sarothrae and Artemisia tridentata samples which included both green and dead
tissue. Samples included flowering parts as well as vegetative portions.
Chemical analysis was performed for both sulfur and fluoride.
1976 VEGETATION COLLECTIONS
Species Studied for Sulfur Accumulation
In April, ZAPS I was surveyed visually for sampling purposes to determine
species present on the plots and their relative abundance and frequency. This
survey was used to select the species to be studied. Those finally chosen were:
SCIENTIFIC NAME
Agropyron smithii
Koeleria cristata
Aristida longiseta
Poa sandbergii (secunda)
Achillea millefolium
Artemisia frigida
Tragopogon dubius
Stipa viridula
COMMON NAME
Western Wheatgrass
Prairie Junegrass
Red Threeawn
Sandberg's Bluegrass
Yarrow
Fringed Sage
Yellow Salsify
Green Needlegrass
T. dubius was collected in early July. ZAPS II was not surveyed so intensively.
A. longiseta was less frequently found so S. viridula was collected instead.
Collection Methods at ZAPS Sites
Pipes delivering the sulfur dioxide and ambient air mixture to the
individual treatment plots were laid out in a pattern used to form the basis for
400
-------
a sampling grid (Figure 13.1). The pipe junctions were used as 25 sampling
points. Each junction was divided further into four quadrants:
NW NE
SW
SE
Two digit random numbers were drawn (00 to 99) and divided by four. The whole
number obtained gave the junction number and the remainder gave the quadrant (1
= NE, 2 = SE, 3 = SW, 0 = NW) to be sampled.
random number -f 4 = junction + quadrant
37 -r 4 = 9 + 1 (9-NE)
88 -r 4 = 22 + 0 (22-NW)
etc.
Quadrants falling north of junctions 1, 11, and 21 were considered outside
the treatment plot, and their respective south quadrants were used for sampling
(i.e., 1-NW, 11-NW, and 21-NW became 1-SW, 11-SW, and 21-SW; 1-NE, 11-NE, and
21-NE became 1-SE, 11-SE, and 21-SE, respectively). Quadrants falling south of
junctions 5, 6, 15, 16, and 25, were similarly rotated to their respective north
quadrants to place them in the treatment plot.
This procedure divided each treatment plot into 86 subplots for vegetation
sampling. A central sampling point was established in each "junction-quadrant"
(subplot) by running a line at a 45° angle from each junction into the appro-
priate quadrant. The line was of a fixed length randomly determined for each
collection period within the restraints of each research team's utilization of
the treatment plots. Appropriate species closest to this point were then col-
lected. No samples were collected within one meter of the pipe.
Ten subplots were chosen for each treatment plot during each collection
period. Material was collected from three separate plants for most species in
each subplot, resulting in one combined sample of each species derived from 30
individual plants. Aristida longiseta was collected from two plants per sub-
plot, giving a sample derived from 20 individual plants. Artemisia frigida was
less frequently found, and the sample was derived from 6 to 29 individual plants
depending on the sample period.
Samples were separated by subplots during early April for Agropyron smithii
and early July and mid-September for both A. smithi i and K. cristata. This
provided ten samples for each type of material (below-ground parts, dead and
live tops)- per species on each treatment plot. Each sample was composed of
material derived from a minimum of three plants in each subplot.
401
-------
un
CO
ro
T
£
ll
^^^^H
••VM
1-SE
5-NE
9-NE
7-NE
12-NE
•Bla^a
^^^
14-NE
19-NE
taHM
17-NE
25-NW
21 -SW
•••^•^
MIXING SHED
MIXING SHED
Figure 13.1A. ZAPS plot sampling junction
numbers.
Figure 13. IB.
Agropyron smithii phenology
subplot locations.
-------
Tops consisted of vegetative culms only for A. smithii, K. cristata, and S.
viridula; both vegetative and reproductive culms for £. sandbergii and Aristida
longiseta, and leaves only for Artemisia frigida and Achillea millefolium. T.
dubius samples were split into leaves, stems, and taproots. Top material was
classified as live or dead based on color (green = live, non-green = dead).
Parts of partially necrotic leaf blades were placed in appropriate live or dead
categories. Cocking (1973) in a field S02 fumigation study used a similar
classification of live and dead.
Below-ground parts for A. smithii consisted of roots, rhizomes, and crowns
to a depth of 15 cm. Weaver (1958) reported 55 percent of the root weight of A.
smithii was found in the top 15 cm of soil. Jorgensen (1970) observed similar
root patterns in central Montana. Below-ground parts of K. cristata consisted
of roots and crowns also collected to a depth of 15 cm. Below-ground parts will
be referred to as roots. Roots were washed with tap water to remove soil prior
to preparation for sulfur analyses.
There were ten sample periods on ZAPS I from early April through mid-
October and nine sample periods on ZAPS II from early April through mid-
September. Sampling was done at approximately two-week intervals. Sulfur
analysis was performed with the Leco induction furnace.
Collection Methods at Drift Sites
Forty marked subplots were located in the area adjacent to the treatment
plots on ZAPS I (Figure 13.2). These will be referred to as the drift plots. A.
smithii was collected in mid-April, mid-July, and mid-September. The mid-April
A. smithii collections included dead tops only. K. cristata was collected in
mid-July and mid-September.
Phenology of Agropyron smithii
In mid-May, 50 individual culms of A. smithii were tagged and numbered on
each treatment plot of ZAPS I and ZAPS II. The culms were arranged in ten groups
of five on each treatment plot. These groups were located in the following
subplots (Figure 13.IB):
1-SE 14-NE
5-NE 17-NE
7-NE 19-NE
9-NE 21-SW
12-NE 25-NW
The culms were marked with a numbered staking flag two inches north of the
respective culms. Each culm was encircled by a twist-tie which led to the base
of the flag. It is felt that neither the staking flag nor the twist-tie
interfered with the growth of the culm. Jorgensen (1970) utilized a similar
403
-------
100 METERS
o
-p.
Figure 13.2. ZAPS I drift plot locati-
ons.
-------
method of marking individual A. smithii culms and also concluded the effect was
negligible.
The specific culms were selected by measuring five feet north from the
appropriate junction along the pipe and then five feet east (west on subplots 21
and 25). The A. smithi i culm closest to this point was #3 for that subplot.
The culm closest to a point six inches west of this was #2, and 12 inches west
was #1. Similarly #4 and #5 were selected on the east side of #3. This was
repeated ten times on each treatment plot.
Data collected for the first period (mid-May) were the total number of
emergent leaves, excluding totally necrotic basal leaves. The twist-tie was
placed on the adaxial side of the highest totally necrotic blade if any were
present. Subsequent inspections involved recording the emergent leaf-blade
number and the number of total necrotic leaf-blades on each culm at approxi-
mately two-week intervals. Sampling was discontinued in mid-September. There
were eight observation periods on ZAPS I and seven observations on ZAPS II.
Seed Collection and Viability Studies
Seeds of A. smithii, K. cristata, £. sandbergii, T. dubius, and S. viridula
were collected from each treatment plot on ZAPS I and II. Seeds of A. smithii
and T. dubius were also collected from a similar vegetation community
approximately 1.6 km southwest of the ZAPS sites.
Each seed collection was pre-treated according to the technique utilized by
Eddleman (1977) for each species to enhance germination. Tetrazolium staining
was conducted on the seed collections to provide seed viability information
according to the rules of the Association of Official Seed Analysts (1970).
Germination tests were conducted to determine the percent germination per
gram weight of seeds collected. Weights of 100 seeds were calculated to adjust
the gram weights in the event that seed size is reduced as a result of the S02
treatments. Two millimeters of hypocotyl growth were considered successful
germination. Seed germination was checked on a daily basis for 30 days. From
this information, an "energy peak" and percent germination was determined for
each seed sample. Analysis of variance and Duncan's (1955) multiple range test
are being conducted on the results.
Seed germination and viability investigations are not far enough along at
this time to present results or discussion on this portion of the 1976 ZAPS
studies.
RESULTS
1975 SULFUR AND FLUORIDE ACCUMULATION AT ZAPS I
On Table 13.1 are the sulfur and fluoride concentrations in foliage of ten
species of grasses, forbs, and shrubs collected from the ZAPS I plots during
1975. Only two grass species (i.e. , A. smithii and B. japonicus) were collected
during both the June and September collection periods. Sulfur levels in these
two grass species increased four times in A. smithii and three times in B.
405
-------
TABLE 13.1. ZAPS PPM SULFUR (S) AND PPM FLUORIDE (F) LEVELS FOR 1975.
ZAPS I
Agropyron smithii
Koeleria cristata
Achillea millefolium
Bromus japonicus
Poa pratensis
-pa
• — >
CTl
ZAPS I
Agropyron smithii
Bromus japonicus
Aristida longiseta
Artemisia tridentata
Bromus tectorum
Stipa comata
Gutierrezia sarothrae
28 June 1975 (31
A (CONTROL) B (LOW)
S F S
1200
800
600
1175
600
3.6
4.6
3.2
2.7
1.4
28 September
A (CONTROL]
S F
850
700
700
1300
400
900
1700
1.0
2.5
0.8
1.6
0.5
0.9
2.4
1500
1050
900
1200
1100
1975 (1
B (LOW)
S
1800
1600
1700
1200
1500
1800
1500
Days Fumigation)
C (MEDIUM)
F S F
2.
3.
3.
4.
2.
23
1.
1.
1.
2.
0.
1.
1.
6
9
3
2
8
Days
F"
4
0
6
4
5
0
9
1600
1200
1200
2200
1200
2.7
2.0
2.3
2.9
2.7
Fumigation)
C (MEDIUM)
S F"
3400
2400
1300
1400
2100
2800
2950
0.8
0.4
1.7
1.8
1.7
1.4
1.0
D (HIGH)
S
2000
800
2200
2550
1400
D (HIGH)
S
5250
3400
3300
2750
4150
2800
5300
1
1
3
2
1
0
1
0
1
0
0
1
F"
.8
.3
.5
.8
.9
F~
.8
.1
.1
.2
.8
.8
.3
ZAPS II Pre-Treatment
S F
900 0.8
600 1.3
800 1.2
1200 2.1
400 1.5
800 0.1
2200 1.8
-------
japonicus, while the fluoride levels decreased during this 90-day period.
Sulfur levels in foliage of all nine species collected during the latter part
of September at the four fumigation concentrations demonstrated that sulfur
accumulation correlated with fumigation concentration and duration of exposure.
Because the vegetation species on the ZAPS I plot survived remarkably well
during the 1975 growing season after continuous fumigation with S02, a large
portion of the 1976 studies was redirected from the ponderosa pine-skunkbush
sites to the ZAPS I and II sites.
1976 SULFUR ACCUMULATION AT ZAPS I AND II
Data analysis of sulfur accumulation in above-ground vegetation from the
ZAPS sites does not include the ninth (mid-September) and tenth (mid-October)
1976 collection periods. However, the analysis covers the major portion of the
growing season. This is confirmed by the 1976 phenology studies on A. smithii
and the 1975 phenology studies conducted by personnel of Colorado State
University (Dodd et aj. , 1976) for P_. sandbergii, K. cristata, Aristida
longiseta, A. smithi i, T. dubius, and Achillea mi 1lefolium. Dormancy is not
induced in Artemisia frigida until later in the season, as is typical of the
sages, because this species is able to extract water from greater depths. In
the presence of adequate fall rains, many prairie species resume growth.
Sulfur levels in live and dead tops for the six species collected throughout
the study period are presented in Appendix A.
The changes in sulfur levels through time were analyzed by fitting a
linear regression to the data by least squares. An initial peak in the sulfur
concept of live tops from the first collection period was observed. This
spring green-up peak was noted in all species. To allow preliminary use of a
simple linear model, this initial collection was excluded from the calculation
of subsequent live regression lines. The coefficients for the regression lines
are presented in Table 13.2. Of the 80 regression coefficients examined, 38
were found to be significant at p ^.10 (Table 13.3).
The number of significant regressions increased with higher levels of
fumigation. Although only two of the regressions on the A plots were signif-
icant, 18 of the 20 regressions showed a positive slope (p <.001), suggesting a
trend of increasing sulfur levels in vegetation on the control plots from the
beginning to the end of the growing season. There was a substantial increase
in the average value of the Coefficient of Determination (R2) from the A plots
(.349) to the B plots (.635). The increase from the B's (.635) to the C's
(.683) and subsequently from the C's to the D's (.743) was less. This
degeneration of the increase in the goodness of fit with higher fumigation
levels was due to the fact that many species reached a marked plateau of sulfur
accumulation in late June through July. This phenomenon will be examined when
individual species are considered.
The regression lines for live tops for six species are presented in
Figures 13.3 through 13.8 (regression was not computed for P. sandbergii
because it was collected during only three sample periods). The trends in
sulfur accumulation can be quickly summarized from these graphs. Sulfur levels
in live vegetation increased with exposure time. The rate of increase rose
407
-------
TABLE 13.2. REGRESSION COEFFICIENTS FOR SULFUR LEVELS THROUGH TIME.
Plot
Z1A
Z2A
Z1B
Z2B
Z1C
Z2C
Z1D
Z2D
Plot
Z1A
Z2A
Z1B
Z2B
Z1C
Z2C
Z1D
Z2D
* =
t = .
t = p
b
o
288
924
250
992
102
120
-792
1410
bo
479
660
-218
307
5
219
-19
-1110
05 < p
01 < p
' < .01
bl
3.27
-0.13
6.00
2.06
9.96
10.20
18.53
11.19
bl
1.37
0.25
8.38
4.38
10.86
10.08
16.58
22.12
< .10
< .05
Live
F
3
0
44
1
13
13
775
1
Live
F
1
0
18
3
22
9
5
7
Agropyron smithii
s
.943
.011
.500;j;
.501
.633t
.839t
.724t
.813
s
.242
.030
.022!
.927
.64?!
.562t
.394*
.257t
R2
.441
.002
.889
.231
.731
.735
.994
.266
Koel
R2
.199
.006
.783
.440
.819
.657
.519
.592
n
7
7
7
7
7
7
7
7
en" a
n
7
7
7
7
7
7
7
7
b
o
17
-15
-433
-287
-917
-1638
-1479
-2346
cristata
b
0
530
382
-288
12
27
-509
-8
-1246
bl
3.90
3.69
8.88
8.97
15.47
20.76
22.50
30.27
bl
0.31
1.10
7.73
5.56
8.17
11.19
11.32
19.66
Dead
F
3.
141.
13.
16.
11.
67.
5.
7.
Dead
F
0.
3.
136.
30.
8.
19.
20.
11.
s
891
767t
503t
006*
231t
929t
731*
909
s
109
315
236t
128t
187*
730t
310*
659*
R2 n
.565 5
.986 4
.818 5
.889 4
.789 5
.971 4
.656 5
.798 4
R2 n
.035 5
.624 4
.978 5
.938 4
.732 5
.908 4
.871 5
.854 4
(continued)
408
-------
TABLE 13.2. (Continued)
Achillea mi lief oil urn
Live
Plot
Z1A
Z2A
Z1B
Z2B
Z1C
Z2C
Z1D
Z2D
Plot
Z1A
Z2A
Z1B
Z2B
Z1C
Z2C
Z1D
Z2D
bo
339
907
542
1223
230
852
-1883
-662
bo
1188
1074
2043
832
1087
-1116
971
-2
bl
1.78
-0.75
2.26
-0.10
8.14
5.75
26.68
25.37
Li
bl
0.91
1.19
-1.70
4.13
7.63
19.41
18.29
23.01
F
1
0
1
0
16
1
33
12
ve
F
0
0
0
1
3
58
6
26
s
.496
.351
.208
.001
.306t
.468
.4901
.81 Of
5
.291
.260
.672
.841
.428
.2771
.890*
.140t
R2
.230
.066
.195
<.001
.765
.227
.870
.719
Artemi
R2
.068
.049
.199
.274
.461
.936
.633
.867
n
7
7
7
7
7
7
7
7
sia
n
6
7
7
7
6
6
6
6
bo
634
494
456
298
410
-831
-451
-2228
frigida
bo
964
680
700
758
1087
-745
1860
-1989
bl
0.13
0.75
3.65
4.91
9.28
16.25
26.64
32.57
bl
1.41
0.89
4.45
2.83
7.63
16.00
9.05
29.78
Dead
F
3.138
2.194
1.035
8.253*
4.173
4.921
6.753*
14.6191
Dead
Fs
102.084*
0.198
10.197
20.120
12.228
9. 125
8.253
4.979
R2
.007
.422
.341
.733
.582
.621'
.692
.830
R2
.990
.163
.911
.953
.461
.901
.892
.833
n
5
5
4
5
5
5
5
5
n
3
3
3
3
3
3
3
3
* = .05 < p < .10
1 = .01 < p < .05
1 = p < .01 (continued)
409
-------
TABLE 13.2. (Continued)
Aristida longiseta
Plot
Z1A
Z1B
Z1C
Z1D
Plot
Z2A
Z2B
Z2C
Z2D
b —
0
R2 -
H :
0
b bn
o 1
-238 4.81
-654 7.99
1473 -0.21
112 9.44
b b,
0 1
577 2.20
673 3.21
479 9.03
1669 11.33
Y-Intercept,
Live
F R2
s
0.926 .481
33.624 .971
<.001 <.001
4.928 .831
Sti
Live
Fs R2
2.379 .373
0.914 .186
3.869 .492
0.812 .169
b-| Slope, FS
Coefficient of Determination,
p = 0, F
n-2]'
n b
0
3 207
3 455
3 191
3 -41
pa viridula
n b
0
6 348
6 -920
6 -1110
6 -1938
Mean Square
Unexpl
b
2
2
5
8
b
2
9
15
24
Dead
F R2
1 s
.19 3.151 .612
.74 132.9941 .985
.82 19.1061 .905
.32 118.4711 .983
Dead
F R2
1 s
.16 4.357 .685
.87 75.000* .987
.75 66.3941 -971
.11 116.062 1 .983
n
4
4
4
4
n
4
3
4
4
due to Regression
ained
n = number of col
Error '
lection periods
* = .05 < p < .10
t = 01 < p < .05
| = p < .01
TABLE 13.3. SUMMATION OF NUMBER OF SIGNIFICANT REGRESSION COEFFICIENTS.
Treatment
Live
Dead
A
0
2
B
2
7
C
6
7
D
7
7
410
-------
6000 -r
5000 -
4000
a
oo
s:
Q.
3000 ~
2000 -
1000
ZAPS I
ZAPS II
90
120
HIGH
T
150
I 1
180 210
JULIAN DATE
I
240
T
270
I
300
Figure 13.3. Least squares, Agropyron smithii, live collections 2 thru 8, ZAPS I and II.
-------
ro
6000 -
5000 -
4000 -
3000 H
Q:
Q
rp
oo
g- 2000
1000 -
90
ZAPS I
ZAPS II
I
120
I
150
I I
180 210
JULIAN DATE
I
240
270
I
300
Figure 13.4. Least squares, Koeleria cn'stata. live collections 2 thru 8, ZAPS I and II.
-------
oo
6000 -I
5000 -
4000 -
g 3000
CL,
Q.
2000 -
1000 H
HIGH
X
X
ZAPS I
ZAPS II
90
I
120
150
I
180
I
210
I
240
I
270
I
300
JULIAN DATE
Figure 13.5. Least squares, Achi1 lea millefolium, live collections 2 thru 8, ZAPS I and II.
-------
6000 -,
5000 -
cc
4000 ~
3000 -
2000 -
1000 -
90
120
150
1 T
180 210
JULIAN DATE
r
240
T
270
1
300
Figure 13.6. Least squares, Artemisia frigida, live collections 2 thru 8, ZAPS I and II.
-------
-pa
cn
6000 -i
5000 -
4000 -
a 3000 -|
co
Q-
QL.
2000 -
1000 -
90
ZAPS I
120
I
150
HIGH
! i
180 210
JULIAN DATE
240
I
270
\
300
Figure 13.7. Least squares, Artisti'da longiseta, live collections 6 thru 8, ZAPS I.
-------
O1
6000 -.
5000 -
4000 —
o;
3000 -
2000 -
1000 -
ZAPS II
HIGH
MEDIUM
LOW
CONTROL
90
120
150
—i r
180 210
JULIAN DATE
—r
240
270
—I
300
Figure 13.8. Least squares, Stipa vin'dula, live collections 3 thru 8, ZAPS II.
-------
with increasing levels of fumigation. Rate of accumulation and total sulfur
were approximately similar on ZAPS I and ZAPS II. The rate of accumulation on
ZAPS II treatment plots D and C tended to be higher than on their respective
ZAPS I counterparts. The rate of accumulation on ZAPS II treatment plots B and
A tended to be lower than on their respective ZAPS I counterparts.
Agropyron smithii
The observed sulfur contents and their regression lines for A. smithii,
live and dead, at the various treatments are plotted in Figures 13.9 through
13.12. At treatment plot A fumigation levels the sulfur content of dead plant
material remained below that of live tissue throughout most of the growing
season. Sulfur levels in the dead material slowly approached the live material
levels and can be expected to exceed these only in late summer as the grass
comes to a full cure. As the intensity of fumigation increased, the sulfur
level in dead tissue exceeded that in live tissue at an earlier date. This was
indicated by both the measured sulfur content and their fitted regression.
Initial sulfur levels in dead tissue were higher on ZAPS I than on ZAPS II; the
sulfur content of live tissue was also higher on ZAPS I, except in vegetation
from the control plots.
A plateau in the sulfur accumulation by live tissue was reached by late
July, except on ZAPS I, plot D, where the rate of accumulation remained linear.
This plateau occurred at a maximum sulfur level of approximately 4,000 ppm.
Sulfur levels in dead tissue peaked as high as 5,675 ppm on the D fumigation
plots. On May 31 A. smithi i was collected from small areas immediately
adjacent to the gas outlets on ZAPS II, plot D. At this time these burn areas
were between 45 and 60 cm in diameter. The A. smithi i collected here displayed
bifacial necrotic lesions coalescing to form interveinal streaks; the streaks
were ivory to light tan. Tingey et aj. (1975) has described these symptoms in
A. smithii treated with 150 pphm S02 in four-hour chamber fumigations. The A.
smithii from these burn areas on ZAPS II, plot D contained 3,350 ppm sulfur.
Similar symptoms were observed on ZAPS II, plot C but the sulfur level in the
tissue was not determined specifically for A. smithi i because individual
species were not separated in this sample.
Koeleria cristata
The responses of K. cristata to the treatments are graphed in Figures
13.13 through 13.16. The relationship between sulfur levels in live and dead
foliage was markedly different from that which occurred for A. smithii. With
J(. cristata, the sulfur content of dead tissue did not exceed that of live
tissue based on the regression analysis; when individual data points were
considered, sulfur levels in live material were exceeded only rarely by levels
in dead tissue. Sulfur levels in dead tissue can be expected to approach those
in live tissue as K. cristata comes to a full cure. Sulfur levels for all
tissue were similar in K. cristata and A. smithii, with a maximum of
approximately 5,000 ppm and a plateau developing later in July.
A comparison of initial sulfur levels in dead K. cristata showed a
residual effect similar to that of A. smithi i. Sulfur levels in live K.
cristata were higher at ZAPS I, plots A and B than at ZAPS II, plots A and B.
417
-------
a;
CL
Q.
6000 -H
5000 -
4000 -
3000 -
2000 -
1000 -
ZAPS I
LIVE •_
DEAD D
90 120 150 180 210 240 270 300
o
cc.
Q
6000 •
5000 .
4000 -
£ 3000 -
00
CL.
CL.
2000 ~
1000
0
LIVE •_
DEAD a
ZAPS II
D
90
~T—
120
—I"
150
T—
180
—I—
210
1
240
1
270
—I
300
Figure 13.9.
JULIAN DATE
Linear regressions of sulfur levels through time in
Agropyron smithii at CONTROL fumigation.
418
-------
C£
6000 -
5000 -
4000 -
3000 -
n_
o_
00 2000
1000
0
LIVE •_
DEAD D
ZAPS I
D
•
90
120 150 180 210 240 270 300
Q
Q-
d.
6000 -.
5000 -
4000 _
3000 -
2000 -
1000
0
ZAPS II
LIVE
DEAD
1 1 1 1— 1 1 1
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.10. Linear regressions of sulfur levels through time in
Agropyron smithii at LOW fumigation.
419
-------
6000 -•
5000 _
E 4000 -
UJ
QL
Q
DC
n_
Q-
3000 -
2000 -
1000 -
0
ZAPS I
LIVE ^_
DEAD n
i r i i i i •
90 120 150 180 210 240 270 300
Q-
Q.
6000
5000 _
4000 -
3000 -
2000 -
1000
0
LIVE
DEAD
ZAPS II
Figure 13.11
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Aqropyron smithii at MEDIUM fumigation.
420
-------
CtL
CL.
Q.
6000 __.
5000 _
4000 -
3000 -
2000 -
1000 -
ZAPS I
LIVE •_
DEAD a
90 120 150 180 210 240 270 300
6000
5000 -
4000 —
a 3000
00
a.
Q-
2000
1000
0
ZAPS II
LIVE •
DEAD n
Figure 13.12.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Agropyron smithii at HIGH fumigation.
421
-------
6000 -i
5000 -
t—
zc —
5 4000 —
Q
CO
CX
3000 _
2000 _
1000 _
ZAPS I
LIVE
DEAD
• M.
90 120 150 180 210 240 270 300
6000 -,
5000 „
\—
re
3 4000 _
Q
CCL
O-
Q-
3000 _
2000
1000
0
LIVE
DEAD
ZAPS II
—r——i 1 1 1 1 1
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.13. Linear regressions of sulfur levels through time in
Koeleria cristata at CONTROL fumigation.
422
-------
ce.
Q
o:
u_
CO
o_
o_
6000 -I
5000 -
4000 -
3000 -
2000 -
1000 -
ZAPS I
LIVE •
•••
DEAD D
D
90
I
120
I
150
180
T
210
i
240
270 300
6000 -i
5000
CD
Hi 4000
a:
a
CsL
GO
D_
Q_
3000
2000
1000 -
LIVE •_
DEAD a
ZAPS II
Figure 13.14.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Koeleria cristata at LOW fumigation.
423
-------
CtL
n.
o.
6000 -,
5000 _
4000 -
3000 -
2000 -
1000 -
0
6000
5000
ZAPS I
LIVE •_
DEAD D
90 120 150 180 210 240 270 300
CD
uj 4000 -
cc.
Q
o:
oo
0.
CL.
3000 -
2000
1000
0
ZAPS II
LIVE •_
DEAD n
Figure 13.15.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Koeleria cristata at MEDIUM fumigation.
424
-------
ex.
D_
6000 -,
5000 -
4000 -
3000 -
2000 -
1000 -
0
ZAPS I
LIVE •_
DEAD a
1 1 1 r 1 1 1
90 120 150 180 210 240 270 300
CD
CXL
Q.
D-
6000 -H
5000
4000 -
3000 -
2000
1000
0
ZAPS II
LIVE •_
DEAD a
Figure 13.16.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Koeleria cristata at HIGH fumigation.
425
-------
Levels in K. cristata were lower initially at ZAPS I, plots C and D than at the
same ZAPS ~II plots, and the green-up sulfur levels at these ZAPS I sites were
among the lowest observed in K. cristata.
Achillea millefolium
Initial sulfur levels for A. millefolium, live and dead, were higher on
ZAPS I, plots B, C, and D than on the pre-treatment ZAPS II plots (Figures
13.17 through 13.20). On ZAPS I, plots B, C, and D, the initial dead levels
exceeded the live. This relationship continued throughout the sample period.
On ZAPS II the sulfur levels in dead leaves did not exceed the live leaves
until later in the season. A July plateau was observed, but the rate of
accumulation on the D plots again began to increase. Maximum values in excess
of 6,000 ppm were observed in both live and dead tissues.
Artemisia frigida
The observed levels and regressions for A. frigida are presented in
Figures 13.21 through 13.24. Sulfur levels in the live leaf material of this
species exceeded that of the dead material, and this relationship continued
throughout the period covered. A residual S02 effect was evident on ZAPS I and
very pronounced on the C and D plots. The plateau was less distinct for A.
frigida; the process of accumulation continued until later in the growing
season on the higher fumigation plots.
Baseline levels for A. frigida were the highest of any of the species that
have been monitored continuously on the ZAPS plots. The baseline level
(treatment A) was in excess of 1,000 ppm and slowly increased throughout the
season with peaks of as high as 1,550 ppm. This high baseline made a
substantial contribution to the higher levels observed throughout the year.
Maximum peaks for live and dead material appeared to be in the 5,000 to 6,000
ppm range and were reached by early July on the D treatment plots. The rate of
accumulation was higher on the ZAPS II plots.
Aristida longiseta
A. longiseta was available in sufficient quantities for sampling only on
ZAPS I. These data are portrayed in Figures 13.25 and 13.26. The
relationship between live and dead was typical of the other perennial
bunchgrasses studied (K. cristata and S. viridula). Live levels exceeded those
of the dead throughout the study period. A residual S02 effect was seen in the
dead samples collected before the 1976 fumigation was started. Live material
could not be collected until early July because A. longiseta is a warm season
grass.
Stipa viridula
S. viridula was collected only on ZAPS II. Live sulfur levels were
generally in excess of those in the dead tissue (Figures 13.27 and 13.28), but
the dead approached the live at an increasing rate as the fumigation level
increased. Live tissue reached a mid-July plateau on the plots under
intentional fumigation. This was followed by a decline in sulfur until mid-
426
-------
6000 _
5000 -
:n
CD
LU 4000 -
C£.
Q
CO
Q_
CL.
3000 -
2000 ~
1000 -
LIVE •_
DEAD n
i i »i i i i
90 120 150 180 210 240 270 300
6000 -,
5000 -
4000 -
3000 -
2000 -
1000 __
ZAPS II
LIVE •_
DEAD n
i i i i 1 n 1
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.17. Linear regressions of sulfur levels through time in
Achillea millefolium at CONTROL fumigation.
427
-------
6000 -i
5000
i—
CD
§ 4000 -I
>-
Q
^ 3000 J
CO
o_
(X
2000 _
1000
0
LIVE •_
DEAD a
ZAPS I
90 120 150 180 210 240 270 300
6000 -,
5000 -
oz
CD
4000 ~
o: 3000 -
o;
Q
Q-
D.
2000 _
1000 „
0
LIVE ^_
DEAD a
ZAPS II
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.18. Linear regressions of sulfur levels through time in
Achillea millefolium at LOW fumigation.
428
-------
CD
D_
Q.
6000 _
5000 _.
4000 _
3000 -
2000 -
1000 -
LIVE •
DEAD a
ZAPS I
D
•
90 120 150 180 210 240 270 300
6000 _
5000 _
~ 4000 J
a:
O
o.
3000 -
2000 -
1000 -
ZAPS II
LIVE
DEAD
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.19. Linear regressions of sulfur levels through time in
Achillea millefpliurn at MEDIUM fumigation.
429
-------
Q.
Q-
6000 -,
5000 —
4000
3000 •
2000
1000
0
D
90 120 150 180 210 240 270 300
ID
OO
D.
CL.
6000 -,
5000 -
4000 -
3000 -
2000 -
1000 _
ZAPS II
LIVE
DEAD D
i i i i i i I
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.20. Linear regressions of sulfur levels through time in
Achillea mi liefoil urn at HIGH fumigation.
430
-------
CD
Qi
Q-
CL.
6000 —
5000 _
4000 -
3000 _
2000 -
1000 -
0
LIVE _•_
DEAD n
ZAPS I
•
"a
a-
90 120 150 180 210 240 270 300
6000 -,
5000 .
i—
m
S 4000 _
o:
Q_
Q-
3000 -
2000 -
1000 -
ZAPS II
LIVE •_
DEAD D
Figure 13.21.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Artemisia frigida at CONTROL fumigation.
431
-------
6000 —
5000 -
£2 4000 _
UJ
o:
CSL
D.
Q_
3000 —
2000 _
1000
0
ZAPS I
LIVE
DEAD
90 120 150
—I T~
180 210
-T 1 1
240 270 300
LU
OL
Q
Q_
Q.
6000 -
5000 -
4000 ~
3000 -
2000 -
1000 —
LIVE
DEAD
ZAPS II
• a
a-
Figure 13.22.
1 1 1 1 1 1 1
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Artemisia frigida at LOW fumigation.
432
-------
6000 -r
5000
o
5 4000
C£.
Q
00
D-
D-
3000
2000
1000
0
LIVE •_
DEAD D
ZAPS I
I I i I i i i
90 120 150 180 210 240 270 300
6000 ~
5000 -
o
£ 4000 _
Q
DS -3000
oo
CL.
Q.
2000
1000
0
ZAPS II
LIVE
DEAD D
Figure 13.23.
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Artemisia frigida at MEDIUM fumigation.
433
-------
CZ3
o:
C/O
D-
Q.
6000 -i
5000 -
4000 -
3000 -
2000 _
1000 -
ZAPS I
90 120 150 180 210 240 270 300
IS)
D-
Q-
6000
5000 _
4000 -
3000 -
2000
1000
0
ZAPS II
LIVE
DEAD
D
Figure 13.24.
1 1 r- 1 1 —T r
90 120 150 180 210 240 270 300
JULIAN DATE
Linear regressions of sulfur levels through time in
Artemisia frigida at HIGH fumigation.
434
-------
Q
C£
OO
D-
Q.
6000 -,
5000 _
4000 -
3000 -
2000
1000
0
ZAPS I — CONTROL FUMIGATION
LIVE •
DEAD a
90 120 150 180 210
240
270
300
o;
Q
oo
a.
a.
6000 -i
5000 -
4000 -
3000 -
2000 -
1000 -
0
ZAPS II —LOW FUMIGATION
LIVE ^_
DEAD n
270
300
Figure 13.25.
90 120 150 180 210 240
JULIAN DATE
Linear regressions of sulfur levels through time in
Aristida longiseta at CONTROL and LOW fumigations.
435
-------
6000 _
5000 _
i— ~
:DC
2 4000 -
UJ
3
>-
° 3000 -
a:
^>
u_
_i
^ 2000
1000
0
D-
O.
ZAPS I—MEDIUM FUMIGATION
LIVE •_
DEAD a
n
90 120 150 180 210 240
270 300
6000
ZAPS II--HIGH FUMIGATION
a:
a
Q-
Q-
5000 _
4000 -
3000 -
2000 -
1000 -
0
LIVE •
DEAD a
--a
i
1
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.26. Linear regressions of sulfur levels through time in
Aristida longiseta at MEDIUM and HIGH fumigations.
436
-------
O-
o.
6000 .
5000
4000 _
3000 -
2000 -
1000 -
ZAPS II--CONTROL FUMIGATION
LIVE
DEAD
90 120 150 180 210 240 270 300
OL
Q
6000 -,
5000 -
4000 -
3000 -
2000 -
1000 -
ZAPS II —LOW FUMIGATION
LIVE •_
DEAD a
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.27. Linear regressions of sulfur levels through time in
Stipa viridula at CONTROL and LOW fumigations.
437
-------
01
Q
ZD
in
6000 -r
5000 _
4000 -
3000 -
2000
1000 -
0
ZAPS II—MEDIUM FUMIGATION
LIVE
DEAD
90 120 150 180 210 240 270 300
6000 -T
5000
i—
E 4000
oo
3000
2000
1000
ZAPS II —HIGH FUMIGATION
LIVE •
DEAD n
T~
90 120
—i r—
150 180
-i 1—
210 240
~I 1
270 300
Figure 13.28.
JULIAN DATE
Linear regressions of sulfur levels through time in
Stipa viridula at MEDIUM and HIGH fumigations.
438
-------
August when the levels rose a second time. Maximum observed level in live
foliage was 4,850 ppm at the D fumigation levels.
Poa sandbergii
The measured sulfur levels for P_. sandbergii are graphed in Figures 13.29
through 13.32. Observed levels in both live and dead tissue increased with the
fumigation level and tended to increase with time. The maximum level in both
live and dead tissue was approximately 2,400 ppm. A limited number of data
points precluded regression computation and interpretation of trends.
Tragopogon dubius
T. dubius was collected only in early July. The measured sulfur levels in
the leaves, taproots, and stems are presented in Table 13.4. The small sample
size precluded testing for significance. T. dubius showed increasing sulfur
levels in all plant parts with higher levels of fumigation. Sulfur levels
decreased in the following order: leaves, stems, taproots. The sulfur levels
in this plant exceeded those in any others that have been studied by the
investigators.
Roots r
A. smithii roots collected in /early April were improperly stored prior to
preparation for sulfur analysis. Molding caused a large reduction in
measurable sulfur levels. These data are not presented. Root collections of
A. smithii and K. cristata made in early July were first tested for homogeneity
of variances (Table 13.5). As the variances could be assumed equal, an anova
was performed (Table 13.6). A significant difference in root sulfur levels
across the treatment plots could not be detected for either species during
early July.
A. smithii and K. cristata roots (ZAPS I and ZAPS II) from mid-September
(September 17, 18, 19; Collection Period 9) displayed equal variances (Table
13.5). Anova indicated significant (p S.05) differences between means for
individual species subjected to the different fumigation levels (Table 13.7).
The observed differences in mean sulfur levels were inspected for significance
(p ^ .05) by the Least Significant Difference (LSD) method (Table 13.8). A.
smithii roots from ZAPS I, plot D had significantly more sulfur than roots from
the other three treatments. On ZAPS II, plot D, sulfur in A. smithii roots
also exceeded levels in roots from the other three treatments. Additionally,
the sulfur content of roots from plot C was significantly greater than those
from plot A. In the case of K. cristata, roots from ZAPS I, plot D, were
significantly higher in sulfur content than roots from the other three
treatment plots. Roots from treatment C also had significantly more sulfur
than those from treatment plot B. K. cristata roots from ZAPS II, plot D, also
had significantly higher sulfur content than those from plots A, B, and C.
Drift Areas
The results of the drift plot collections (ZAPS I) (including
mid-September) are presented in Table 13.9. Several significant differences
439
-------
>_
CD
i— i
UJ
3
>-
OL
Q
ZD
U-
s:
fe
CD
H-4
li 1
3
o;
Q
Qi
§
00
s:
O-
Q_
6000 — j
5000 _
_
4000 _
3000 -
2000 _
1000 _
0 -
9
ZAPS I
LIVE •
DEAD n
• ' • D
! 1 1 1 1 II
0 120 150 180 210 240 270 300
JULIAN DATE
6000 -n
5000 —
4000 -
3000 —
_
2000 _
1000 _
0
c
LIVE • ZAPS II
DEAD n
a
I 1 I 1 1 1 1
10 120 150 180 210 240 270 300
Figure 13.29.
JULIAN DATE
Sulfur levels through time in Poa sandbergii at CONTROL
fumigation.
440
-------
o
UJ
OL
C£
00
Q_
6000 -,
5000 —
4000 —
3000 -
g: 2000 -
1000 —
0
LIVE •
DEAD D
90
1 1
120 150
1
180
1
210
1 1
240 270
300
6000 -
" ^
| 5000 -
i__<
UJ —
3
£ 4000 -
Q
D;
— i
j 3000 -
oo
SI
2: 2000 -
^F—
1000 -
0
ZAPS II
LIVE •
DEAD D
' . • D
i r i ' i i i
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.30. Sulfur levels through time in Poa sandbergi i at LOW
fumigation.
441
-------
UJ
3
a:
a:
rs
LJ_
_i
ZD
00
6000 —i
5000 —
4000 -
3000 -
2000 -
1000 _
ZAPS I
LIVE*
DEADD
I i i i i i '
90 120 150 180 210 240 270 300
6000 —,
H- 5000 _
CD
t—i
UJ
Q
a:
4000 _
3000
2000 -
1000
ZAPS II
LIVE •
DEAD D
Figure 13.31.
i 1 1 1 , ,
90 120 150 180 210 240 270
JULIAN DATE
Sulfur levels through time in Poa sandbergii at MEDIUM
fumigation.
442
-------
6000 —i
5000 —
CJ
I—I
LU
4000 -
£ 3000 _
OL
Q
£ 2000
1000 -
0
LIVE •
DEAD a
ZAPS I
T I I I I I I
90 120 150 180 210 240 270 300
6000 —r
5000 —
4000 _
3000
2000 —I
1000 -
ZAPS II
LIVE •
DEAD a
90 120 150 180 210 240 270 300
JULIAN DATE
Figure 13.32. Sulfur levels through time in Poa sandbergi i at HIGH
fumigation.
443
-------
TABLE 13.4. SULFUR (ppm) IN Tragopogon dubius FROM EARLY JULY
Treatment
Z1A
X
Z1B
X
Z1C
X
Z1D
(col lection
Leaves
4775
4450
3100
4108
6675
5750
7850
7125
8625
5750
7850
7408
5700
9450
period 5)
Taproot
1950
1550
1150
1550
1550
1900
1400
1617
1550
1450
1750
1583
1150
2450
Stem
-
-
850
-
-
-
1825
-
-
2000
-
3150
7575
1800
444
-------
TABLE 13.5. MEAN SULFUR CONTENT AND BARTLETT'S TEST OF HOMOGENEITY OF
VARIANCES FOR ROOTS FROM A. Smithii AND K. Cristata.
Collection Period 5, Early July.
A. smithii
Treatment
Z1A
Z1B
Z1C
Z1D
x ppm
780
720
790
811
df=n-l
9
9
9
8
X2 = 5
Collection Period
A. smithii
Treatment
Z1A
Z1B
Z1C
Z1D
Z2A
Z2B
Z2C
Z2D
x ppm
730
738
720
830
720
775
790
870
df=n-l
9
9
9
9
X2 - 2
9
9
9
9
X2 = 4
S2
9,565
7,327
6,006
1 ,739
.28
x ppm
792
789
805
725
9, Mid-September,
S2 x ppm
3,481
6,889
2,916
6,241
.27
1 ,764
3,969
6,561
7,921
.92
765
725
835
970
738
775
825
935
1976
K. cristata
df=n-l
9
8
9
9
X2 =
1976
K. cristata
df=n-l
9
9
9
9
x2 -
8
8
8
9
X2 =
S2
6,115
11,109
6,368
7,921
0.94
S2
14,400
6,241
13,917
7,921
2. 15
14,884
5,041
6,400
13,924
3.13
Critical Chi-square x2 -05 [3] = 7.82
445
-------
TABLE 13.6. ANOVA OF SULFUR CONTENT OF ROOTS FOR Agropyron smithii AND
Koeleria cristata FROM ZAPS I IN EARLY JULY.
(collection period 5)
Agropyron smithii
Source of Variation Sum of Squares Degrees of Freedom
Treatments 44,470 3
Residual 219,889 35
Total 264,359 38
Critical F Q5[3>35] = 2.875
Koeleria cristata
Source of Variation Sum of Squares Degrees of Freedom
Treatments 38,542 3
Residual 272,451 35
Total 310,993 38
Critical F_05[3)35]= 2.875
Mean Square
14,823
6,283
FS = 2.359
Mean Square
12,847
7,784
F =1.650
446
-------
TABLE 13.7. ANOVA OF SULFUR CONTENT OF ROOTS FOR A. smithi1 AND K. cristata
FROM ZAPS I AND ZAPS II IN MID-SEPTEMBER.
(collection period 9)
Source of Variation
Treatments
Residual
Total
Critical F Q5[3
Source of Variation
Treatments
Residual
Total-.
Critical F;05[3
Source of- Variation
Treatments
Residual
Total
Critical F_05[3
Source of Variation
Treatments
Residual :
Total
Critical F Q5[3
Agropyron smithii ZAPS
Sum of Squares Degrees
77,797
174,562
252,359
,36] - 2-88
Koeleria cristata ZAPS
Sum of Squares Degrees
382,750
347,188
729,938
.36]. = 2'88 :
Agropyron smithii ZAPS
Sum of Squares Degrees
115,188
182,250
297,438
,36] = 2'88
Koeleria cristata ZAPS
Sum of Squares Degrees
202,250
309,000
511,250
= 2 92
,30] ^'^
I
of Freedom
3
36
39
I
of Freedom
3
36
39
II
of Freedom
3
36
39
II
of Freedom
3
30
33
Mean Square
25,932
4,849
F = 5.348
Mean Square
127,583
9,644
F = 10.885
s
Mean Square
38,396
5,063
F = 7.584
Mean Square
67,417
10,300
F = 6.545
447
-------
TABLE 13.8. DIFFERENCES IN SULFUR CONTENT OF ROOTS COLLECTED IN MID-
SEPTEMBER COMPARED BY LEAST SIGNIFICANT DIFFERENCES.
Agropyron smithii ZAPS I
LSD
63
.05
720
C
730
A
738
B
830
D
720
C
10
18
no
730
A
100
738
B
92
830
D
Koeleria cristata ZAPS I
Agropyron smithii ZAPS II
LSD
65
.05
720
A
775
B
790
C
870
D
720
A
55
70
150
775
B
15
95
790
C
*
80
870
D
Koeleria cristata ZAPS II
LSD
104
.05
738
A
775
B
825
C
935
D
738
A
47
87
*
197
775
B
50
160
825
C
110
935
D
''Signif icant at p£. 05
TABLE 13.9. SULFUR LEVELS IN DRIFT PLOT COLLECTION.
Col lection
Date
13 April
16 July
Julian
Date
104
198
Species Material
A. smithii
Dead
Live
Dead
Roots
K. cristata Live
Dead
" Roots
X
472
1004
528
600
802
612
752
Sx
12
36
32
18
44
31
22
40
34
18
18
20
19
20
95%
Con/idence
Limits
L1 Lo
449
933
459
562
711
546
705
496
1075
596
638
894
678
800
448
-------
(p ^ .05) in partitioning of sulfur within the plants and between A. smithii
and K. cristata were observed in the mid-July collection. A. smithii live
tissue had more sulfur than either the dead tissue or the roots. K. cristata
had more sulfur in the live tissue than in the dead. The level of sulfur in
the dead tissue was less than that in the roots. Finally, live A. smithii
contained more sulfur than K. cristata, while the dead tissue and roots of A.
smithii contained less than did K. cristata.
A cluster pattern of differing sulfur levels within the drift areas was
not established by the sampling grid used (Figure 13.5, Materials and Methods).
The levels of sulfur by species in live culms for the entire mid-July drift
collection were compared to the levels predicted by the A and B treatments on
ZAPS I. Regression for both species at the LOW treatment level was significant
and used to establish 95% prediction belts for sample sizes of 34 in the case
of A. smithi and 20 in the case of K. cristata. The linear model for the A
treatment (0 pphm) was rejected, and a prediction belt was constructed on 7 =
Y. The 95% confidence interval for the grasses from the drift plot area was
then compared to the prediction belts for the appropriate point in time (July
16, i.e., day 198). This is portrayed graphically in Figure 13.33. The drift
collections showed significantly lower levels in both species than in grasses
fumigated on the B treatment plot. Although the means were higher than those
from the plot A collections, they were not significantly so. No clusters on
the drift areas were recognized as significantly higher than the plot A
controls.
PHENOLOGY
The results of the phenological observations of the vegetative stages of
A. smithii are summarized in Tables 13.10 through 13.12. Total leaf number
increased rapidly through early July and leveled off by the next observation
period in early August (Tables 13.10 and 13.11). No significant differences (p
^ .05) in total leaf number were observed between the treatment plots at any
observation date (Table 13.12).
The ratio of necrotic leaves to live leaves was examined to determine
differences between treatments on each ZAPS plot during each observation
period. The last row of Table 13.10 places the treatment plots on ZAPS I in
order from that with the highest ratio of necrotic leaves to that with the
lowest ratio of necrotic leaves for each observation period. The only
significant (p i .05) difference observed was the September 2 collection in
which vegetation from ZAPS I, plot D (HIGH) displayed a higher ratio of
necrotic leaves than vegetation from ZAPS I, plot B (LOW). However, the trend
from early July through mid-September was consistent, treatment plot D showing
the highest proportion of necrotic leaves, followed by A (CONTROL), then C
(MEDIUM), and finally B showing the smallest proportion of necrotic leaves.
This relationship is portrayed graphically in Figures 13.34 through 13.36,
where necrotic leaves (expressed as a percent of the total leaf number) of plot
D are compared to plots A, B, and C. The exact probabilities (as opposed to a
^ .05) of obtaining the observed differences on each date are presented in the
next to the last row. The low probabilities from July on are indicative of the
strength of the trend DACB. Only the HIGH treatment showed a higher level of
necrosis than A (CONTROL).
449
-------
2000 -r
CD
Q
co
Q_
O-
1500 4-
1000
500
LIVE AGROPYRON SMITHII
ZAPS
130
2000
1500 --
1000
Q-
O-
500
130
Figure 13.33.
CONTROL
150 170
190
210
230 250
LIVE KOELERIA CRISTATA
ZAPS I ( K=20 )
150 170 190 210
JULIAN DATE
230 250
95% prediction belts for treatment plots and
95% confidence interval ( J ) for drift plot
col lection.
450
-------
TOO -,
80 -
oo
LU
60 .
O
LU
LU
O
LU
Q.
40 -
20 -
ZAPS I D (HIGH)
ZAPS I A (CONTROL)
INDICATES OBSERVATIONS WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
X
X
120
i
140
160
I
180
200
JULIAN DATE
220
240
260
Figure 13.34. Percent necrotic leaves of Agropyron smithii.
-------
en
ro
oo
LU
o
o
o;
o
UJ
a.
100 -,
80 •
60 -
40 •
20-
ZAPS I D (HIGH)
ZAPS I B (LOW)
INDICATES OBSERVATIONS WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
120
140
•
i
160
180 200
JULIAN DATE
220
240
260
Figure 13.35. Percent necrotic leaves of Agropyron smithi i.
-------
100-t
80
oo
LU
ZAPS I D (HIGH) -
ZAPS I C (MEDIUM)-
INDICATES OBSERVATIONS WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
o
^—t
I—
O
60
on
CO
O
LU
Q_
40 -
20-
120
140
i
160
i i
180 200
JULIAN DATE
1
220
240
260
Figure 13.36. Percent necrotic leaves of Agropyron smithii.
-------
TABLE 13.10. Agropyron smithii ZAPS I PHENOLOGY STUDIES AND TEST FOR INDEPENDENCE OF LEAF NECROSIS.
Observation #
Date 17,
7TA
ZIB
ZIC
/1U
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
%Necrotic
RXC Test of
Independence*
1
18 May
147
147
0
0
157
157
0
0
154
154
0
0
151
151
0
0
G
I-P(G)
Maximal Nonsignificant
ae us u v . u j
2
1 Jun
193
161
32
16.6
201
161
40
19.9
198
159
39
19.7
200
162
38
19.0
0.908
.823
B C D A
3
16, 18 Jun
216
175
41
19.0
246
186
60
24.4
234
175
59
25.2
222
169
53
23.9
3.031
.387
C B D A
4
4, 5, 6 Jul
288
196
92
31.9
292
213
79
27.1
259
178
81
31.3
237
146
91
38.4
7.757
.051
DACB
5
2 Aug
310
180
130
41.9
328
204
124
37.8
322
197
125
38.8
301
172
129
42.9
2.305
.512
DACB
6
16 Aug
311
137
174
55.9
331
171
160
48.3
327
167
160
48.9
312
136
176
56.4
7.352
.061
DACB
7
2 Sept
312
112
200
64.1
332
148
184
55.4
327
128
199
60.9
313
100
213
68.1
11.736
.008
DACB
8
17 Sept
312
83
229
73.4
333
112
221
66.4
329
89
240
72.9
313
82
231
73.8
5.894
.117
DACB
*Source: Sokal and Rohlf, 1969.
-------
TABLE 13.11. Agropyron smithi i ZAPS II PHENOLOGY STUDIES AND TEST FOR INDEPENDENCE OF LEAF NECROSIS.
Observation.*
Date
Z2A
Z2D
23,
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
% Necrotic
Total
Live
Necrotic
% Necrotic
RXC Test of
Independence*
1 2
24 May No Sample
159
159
0
0
158
158
0
0
164
164
0
0
145
145
0
0
G
l-P(G)
Maximal Nonsignificant
Sets a < .05
3
18, 21 Jun
201
153
48
23.9
230
184
46
20.0
230
174
56
24.3
220
150
76
31.8
8.578
.035
DCAB
4
6, 7 Jul
275
201
74
26.9
280
213
67
23.9
294
201
93
31.6
289
170
119
41.2
22.444
.001
DCAB
5
3 Aug 1
281
151
130
46.3
292
158
134
45.9
304
161
143
47.0
316
127
189
59.8
16.855
.001
DCAB
6
5 Aug 2
281
137
144
51.2
296
143
153
51.7
307
148
159
51.8
319
124
195
61.1
8.686
.034
D C B A
7
Sept
285
90
195
68.4
302
104
198
65.6
307
102
205
66.8
321
88
233
72.6
4.149
.246
D A C
8
19 Sept
286
45
241
84.3
303
64
239
78.9
308
68
240
11. B
323
57
266
82.4
5.130
.163
B A D B C
^Source: Sokal and Rohlf, 1969.
-------
TABLE 13.12. CHI-SQUARE TEST OF TOTAL LEAF NUMBER AT EACH OBSERVATION.
Col lection
Number
1
2
3
4
5
6
7
8
ZAPS
X2
0.360
0.192
2.314
7.487
1.392
0.982
0.941
1.090
I
E
152
198
230
269
315
320
321
322
ZAPS II
X2
1.259
No
2.546
0.777
2.296
2.609
2.182
2.289
E
156
Sample
220
285
298
301
304
305
X2.05[3] - 7.815; See Tables 13.10 and 13.11 for Observed Total Leaf Values.
On ZAPS II (Table 13.10) the D treatment showed significantly (p S .05)
higher necrosis than the A treatment from early July through mid-August. Plot
D was also significantly higher in necrosis than plot B from late June through
early August. Finally, the D treatment showed significantly more necrosis than
the C treatment in early July. The trend was DCAB until mid-August when it
became less consistent. These data are portrayed graphically in Figures 13.37
through 13.39, where plot D is compared to plots A, B, and C.
DISCUSSION AND SUMMARY
SULFUR
Sulfur levels in live plant foliage of the seven species of grasses, forbs,
and semi-shrubs collected at ZAPS plots during 1976 adequately demonstrate that
elevated concentrations are directly related to the duration of fumigation and
the S02 concentrations delivered at the four treatment levels. The foliage of
the four species (Agropyron smithii, K. cristata, Achillea millefolium, and
Artemisia frigida) which started to grow before fumigations began at ZAPS sites
I and II in April of 1976 showed higher sulfur levels than are normally found
during the rest of the growing season (i.e., average 1,370 vs. 950 ± 75 ppm).
Evidence of residual sulfur concentrations in live tissue resulting from the
1975 fumigation on ZAPS I was demonstrated clearly only in one of the species
(A. millefolium) on the B, C, and D treatment plots, while residual sulfur was
evident in live foliage of A. frigida from the C and D treatment plots. Neither
of the two early season grasses (A. smithii and K. cristata) collected in 1976
demonstrated evidence of elevated residual sulfur concentrations due to 1975
456
-------
100 „
80 -
60 -
-p.
en
O
i—i
o
o
Di
UJ
40 -
20
120
ZAPS ii D (HIGH;)
ZAPS II A (CONTROL)
INDICATES OBSERVATIONS WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
i
140
i
160
220
i
240
180 200
JULIAN DATE
Figure 13.37. Percent necrotic leaves of Agropyron smithii
260
-------
TOO
80
60
§
-fs.
en
00
40
20
ZAPS II D (HIGH)
ZAPS II B (LOW)
INDICATES OBSERVATIONS WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
x
X
X
/•
s
X
X
x
X
x
X
X
120 140
I
160
I
180
I
200
r
220
I
240
JULIAN DATE
Figure 13.38. Percent necrotic leaves of Agropyron smithii.
I
260
-------
cn
100^
80-
S 60-
§
o
o
LU
Q,
40-
20-
o-.
IfO
ZAPS II D (HIGH)
ZAPS II C (MEDIUM)
INDICATES OBSERVATION WHEN THE
DIFFERENCE WAS SIGNIFICANT AT P<.05
/
/
/
I
140
I
160
I
180
JULIAN DATE
I
200
220
I
240
r
260
Figure 13.39. Percent necrotic leaves of Agropyron smithii
-------
fumigation. However, because of the elevated sulfur levels in both of these
grasses during the early flush, any residual sulfur in the roots which could be
translocated up to the foliage could be masked (see Table 13.8, Differences in
Sulfur Content of Roots Collected in Mid-September Compared by Least Signif-
icant Differences). More detailed studies of sulfur levels in the root
tissues of these two species will be undertaken during the 1977 growing season
because it is necessary to understand the role of residual sulfur concentra-
tions if one is to understand and correlate sulfur levels in the foliage being
fumigated and leaf necrosis and/or early senescence.
The need for more detailed studies on the role of residual sulfur also was
made apparent by the sulfur levels found in remnant populations of A. smithii
collected in the winter of 1976 at two air monitoring stations in Anaconda,
Montana. These concentrations are presented in Appendix B, Table B-l, as are
the sulfur contents of A. smithii foliage collected in mid-February (1977) from
the D treatment plot (ZAPS I) one week after collections were made from the
Anaconda sites (Appendix B, Table B-2).
Figure B (Appendix B) relates the sulfur data in this appendix to ambient
S02 concentrations and sulfur content of A. smithi i foliage. The total monthly
dosage of S02 delivered during a 14-hour day length period (6 a.m. to 8 p.m.)
from April 1 to September 30, 1976, at the two Anaconda monitoring sites is
compared to the total monthly dosage for similar day length periods on ZAPS I
at the four treatment levels (basic data for ZAPS sites is from 1975
fumigation, unpublished data, Lewis and Lee, EPA). Dosage was computed from
hourly geometric means of the ambient concentrations. Figure B also shows the
95% confidence intervals for the sulfur content of A. smithii foliage for the
February (1977) collections from ZAPS I, plot D, and the Anaconda sites.
The data on Figure B suggest three explanations for the discrepancies
between S02 air concentrations and sulfur accumulation in the foliage from the
Anaconda sites and the ZAPS I, D treatment plot: (1) The sulfur levels in
foliage from the Anaconda sites are due to residual sulfur in the roots and
rhizosphere of these plants; (2) the remnant population of A. smithii at the
Anaconda sites consists of ecotypes which are S02 tolerant and have adapted to
accumulating excessive levels of sulfur, or (3) the ambient S02 concentrations
measured at either the Anaconda or ZAPS monitoring sites are not revealing the
actual amounts of S02 impacting the foliage of A. smithii. Until the
investigators finish the 1977 ZAPS studies and grow A. smithTi in Anaconda
soils (from Highway Monitoring Site) in the University of Montana Department of
Botany gardens this year, it will not be known whether the forementioned
factors are collectively or singly responsible for the difference in sulfur
accumulation between the ZAPS I, D treatment plot and the Anaconda sites.
PHENOLOGY
The 1976 phenology studies on A. smithii across the ZAPS I treatment plots
demonstrate that this grass species from the control plot (plot A) is possibly
influenced by abiotic factors other than S02. The rate of leaf necrosis on
plot A is more similar to that which occurred on the most heavily-fumigated
plot (plot D) than to either the B or C treatment plots. This similarity could
be caused or partially caused by the fact that the A treatment plot has lower
460
-------
soil moisture (1976 soil moisture data from Dodd and Fort Collins, Colorado,
study group) in the top 30 cm of soil than the D treatment plot. Differences
between soil moisture in the top 30 cm of soil on plots A and D occurred in
late June (June 24) and continued on through the remaining growing season.
While the overall difference was only 19 percent (i.e., 4.39 g/cm2) from June
24 to September 6 (74 days), this measured reduction of soil moisture on
treatment plot A might explain why leaf necrosis at this location is most
similar to the leaf necrosis on treatment plot D (see Table 13.10, Phenology of
ZAPS I).
The soil moisture profile of all four treatment plots on ZAPS I should be
examined thoroughly during the 1977 growing season to ascertain whether there
are significant differences between the plots which could be influencing the
results of phenology, biomass, and species diversity studies being carried out
on ZAPS I.
Phenology studies of A. smithii on ZAPS II (Table 13.11) during the period
of mid-June (Julian Day 170) to mid-August (Julian Day 228) demonstrated that
S02 concentrations on that D plot caused significantly more leaf necrosis than
that occurring on the other three treatment plots. However, the S02
concentrations delivered to the B and C treatment plots did not cause a
significant difference in leaf necrosis from that occurring on the control
plot. This would indicate that the gradient for these concentrations, while
causing increases in sulfur accumulation in foliage over baseline, are not
adequate to cause differences in visible damage to the foliage of A. smithi i.
Phenology studies of A. smithii and one other grass species during the
1977 growing season (possibly K. cristata) will be carried out to compare and
verify the results of the 1976 phenology studies.
REFERENCES
Association of Official Seed Analysts. 1970. Rules for Testing Seeds, 1970.
Proc. Assoc. Official Seed Analysts. 60(2):116.
Cocking, W. D. 1973. Plant Community Damage and Repairabi1ity Following
Sulfur Dioxide Stress on an Old-Field Ecosystem. Ph.D. Thesis, Rutgers
University. 133 pp.
Dodd, J. L. et. aj.. 1976. Effects of S02 and Other Coal-Fired Plant Emissions
on Producers, Invertebrate Consumers, and Decomposer Structure and
Function in an Eastern Montana Grassland. Quarterly Progress Report,
Grant No. A-803176-01. Natural Resources Ecology Laboratory, Colorado
State University, Fort Collins.
Dreisinger, B. R. 1965. Sulfur Dioxide Levels and Effects of the Gas on
Vegetation Near Sudbury, Ontario. Presented at 58th Annual Meeting of the
Air Pollution Control Association. Paper No. 65-121.
Dreisinger, B. R. and P. C. McGovern. 1970. Monitoring atmospheric sulfur
dioxide and correlating its effects on crops and forests in the Sudbury
461
-------
area. Proceedings of the Impact of Air Pollution on Vegetation Confer-
ence, (S.N. Linzon, ed.), Toronto, Ontario.
Duncan, D. B. 1955. Multiple range and multiple F tests. Biometrics. 11:1-
41.
Eddleman, L. E. 1977. Personal Communication. (Rangeland Ecologist, School of
Forestry, University of Montana, Missoula, Montana).
Jorgensen, H. E. 1970. A Life History Study of Agropyron smithii Ryalb in
Central Montana with Related Effects of Selected Herbicide Treatment of
Rangeland. Ecological Effects of Chemical and Mechanical Sagebrush
Control. Progress Report for period ending 6/30/70. Montana Department of
Fish and Game, Game Management Research Bureau, Bozeman, Montana. VM05-R-
3,4,5.
Lee, J. J. and R. A. Lewis. 1976. Unpublished Report. U. S. Environmental
Protection Agency, Corvallis Environmental Research Laboratory.
Linzon, S. N. 1971. Economic effects of sulfur dioxide on forest growth. J.
Air Poll. Control Assoc. 21:81-86.
Tingey, D. T. , R. W. Field, and L. Bard. 1975. Physiological responses of
Vegetation to coal-fired power plant emissions. In The Bioenvironmental
Impact of a Coal-Fired Power Plant, Second Interim Report, Col strip,
Montana—June 1975 (R. A. Lewis, N. R. Glass, and A. S. Lefohn, eds.).
Environmental Research Laboratory, Office of Research and Development,
Corvallis, Oregon. EPA-600/3-76-013.
Weaver, J. E. 1958. Underground development in natural grassland communities.
Ecological Monograph. 28:55-77.
462
-------
APPENDIX A
en
TABLE A-l. ZAPS I - TREATMENT A (CONTROL)
1976 COLLECTION
Sulfur Levels (ppm)
Collection
Date
6 April
13 May
1 June
18 June
4 July
2 August
16 August
1 September
Jul ian
Date
97
134
153
170
186
215
229
245
A. smithii
Live
1300
700
850
1000
700
1000
850
1275
Dead
500
NS
NS
NS
500
900
750
1225
K. cristata
Live
1675
700
550
900
650
800
700
875
Dead
600
NS
NS
NS
550
500
550
750
P. sandbergii
Live
NS
500
550
550
NS
NS
NS
NS
Dead
NS
NS
NS
NS
400
NS
NS
NS
A. longiseta
Live
NS
NS
NS
NS
1000
750
950
900
Dead
400
NS
NS
NS
NS
700
850
600
A. mill
Live
800
600
550
850
550
550
750
900
efolium
Dead
700
NS
NS
NS
550
650
600
800
A. frigida
Live
1100
1250
-
1550
1300
1250
1300
1550
Dead
1100
NS
NS
NS
NS
NS
1300
1300
NS^not sampled
-=missing sample
-------
APPENDIX A
en
TABLE A-2. ZAPS II - TREATMENT A (CONTROL)
1976 COLLECTION
Sulfur Levels (ppm)
Col lection
Date
10 April
8 May
31 May
18 June
6 July
3 August
15 August
1 September
Julian
Date
101
128
152
170
188
216
228
245
A. smithii
Live
1716
800
850
1050
1050
800
950
800
Dead
350
NS
NS
NS
NS
800
850
850
K. cristata
Live
1575
650
750
900
500
650
650
850
Dead
500
NS
NS
NS
NS
550
700
650
P. sandbergii
Live
NS
650
600
700
NS
NS
NS
NS
Dead
NS
NS
NS
NS
450
NS
NS
NS
S. viridula A. mill
Live
NS
800
800
1100
950
1150
1000
1100
Dead
550
NS
NS
NS
NS
950
750
850
Live
1175
850
700
950
650
600
850
750
efolium
Dead
550
NS
NS
NS
650
700
700
600
A. frigida
Live
1600
1550
913
1350
1133
1250
1350
1550
Dead
750
NS
NS
NS
NS
NS
1050
750
-------
APPENDIX A
TABLE A-3. ZAPS I - TREATMENT B (LOW)
1976 COLLECTION
Sulfur Levels (ppm)
Col lection
Date
7 April
13 May
1 June
16 June
4=.
171 5 July
2 August
16 August
2 September
Jul ian
Date
98
134
153
168
187
215
229
246
A. smithii
Live
1400
1050
1050
1350
1450
1500
1700
1650
Dead
600
NS
NS
NS
850
1400
1650
2000
K. cristata
Live
1575
900
900
1425
1200
1850
1550
1800
Dead
500
NS
NS
NS
1050
1400
1550
1600
P. sandbergii
Live
NS
950
783
850
NS
NS
NS
NS
Dead
NS
NS
NS
NS
500
NS
NS
NS
A. longiseta
Live
NS
NS
NS
NS
850
1050
1200
1300
Dead
725
NS
NS
NS
NS
1050
1050
1150
A. mil lefol ium
Live
1275
800
800
1200
700
1200
1150
950
Dead
-
NS
NS
NS
1050
1350
1400
1225
A. frigida
Live
1700
1950
1450
1850
1750
1933
1600
1500
Dead
1150
NS
NS
NS
NS
NS
1600
1900
-------
APPENDIX A
TABLE A-4. ZAPS II - TREATMENT B (LOW)
1976 COLLECTION
Sulfur Levels (ppm)
en
en
Col lection
Date
10 April
11 May
1 June
21 June
7 July
3 August
15 August
2 September
Julian
Date
101
132
153
173
189
216
228
246
A. smith ii
Live
1250
1100
1300
1450
1600
1550
1250
1450
Dead
600
NS
NS
NS
NS
1900
1500
1950
K. cristata
Live
1050
800
1100
850
1300
1550
1050
1350
Dead
550
NS
NS
NS
NS
1350
1250
1300
P. sandbergii
Live
NS
975
800
950
NS
NS
NS
NS
Dead
NS
NS
NS
NS
850
NS
NS
NS
S. viridula
Live
NS
1275
950
1350
1600
1300
1100
1600
Dead
-
NS
NS
NS
NS
1200
1350
1500
A. mi lief oli urn
Live
850
1050
1050
1800
1100
1300
775
1350
Dead
700
NS
NS
NS
1400
1550
1300
1350
A. frigida
Live
2125
1350
1050
1900
1950
1750
1550
1800
Dead
1050
NS
NS
NS
NS
NS
1350
1500
-------
APPENDIX A
TABLE A-5. ZAPS I - TREATMENT C (MEDIUM)
1976 COLLECTION
Sulfur Levels (ppm)
Col lection
Date
7 April
13 May
1 June
16 June
5 July
2 August
16 August
2 September
Jul ian
Date
98
134
153
168
187
215
229
246
A. smithii
Live
1300
1350
1500
2050
1750
2550
2550
2225
Dead
800
NS
NS
NS
1200
2750
2950
2800
K. cristata
Live
1200
1200
1600
2250
2100
2350
2400
2600
Dead
850
NS
NS
NS
1250
2200
2000
1800
P. sandbergii
Live
NS
1150
1450
988
NS
NS
NS
NS
Dead
NS
NS
NS
NS
900
NS
NS
NS
A. longiseta
Live
NS
NS
NS
NS
1450
1250
1750
1275
Dead
750
NS
NS
NS
NS
1400
1700
1500
A. mill
Live
900
1150
1450
1650
2000
2200
1850
2150
efol ium
Dead
1050
NS
NS
NS
2700
2850
2150
2350
A. frigida
Live
2700
2050
2050
-
3100
2700
2350
3150
Dead
1100
NS
NS
NS
NS
NS
2325
2100
-------
APPENDIX A
TABLE A-6. ZAPS II - TREATMENT C (MEDIUM)
1976 COLLECTION
Sulfur Levels (ppm)
-P-
en
co
Col lection
Date
9 April
11 May
31 May
21 June
7 July
3 August
15 August
2 September
Jul ian
Date
100
132
152
173
189
216
228
246
A. smithi i
Live
1100
1150
1700
2200
2150
2616
2200
2450
Dead
400
NS
NS
NS
NS
2850
3400
3200
K. cristata
Live
1675
1550
1300
2250
2450
2650
2250
2550
Dead
550
NS
NS
NS
NS
2250
1950
2050
P. sandbergii
Live
NS
1450
1250
1250
NS
NS
NS
NS
Dead
NS
NS
NS
NS
1100
NS
NS
NS
S. viridula
Live
NS
2500
1600
2250
2600
2100
2250
2950
Dead
450
NS
NS
NS
NS
2500
2250
2800
A. millefolium
Live
550
1400
1200
2550
2200
2350
2150
1800
Dead
500
NS
NS
NS
2450
3750
2800
2250
A. frigida
Live
1250
1650
1750
-
2325
3150
3050
3950
Dead
800
NS
NS
NS
NS
NS
3350
2800
-------
APPENDIX A
-P-
en
TABLE A-7. ZAPS I - TREATMENT D (HIGH)
1976 COLLECTION
Sulfur Levels (ppm)
Collection
Date
8 April
13 May
1 June
16 June
4 July
2 August
16 August
2 September
Jul ian
Date
99
134
153
168
186
215
229
246
A. smithii
Live
1500
1750
1950
2400
2600
3175
3450
3800
Dead
1000
NS
NS
NS
1519
4725
3200
4100
K. cristata
Live
1200
1650
2017
3350
3850
4275
3400
3400
Dead
1050
NS
NS
NS
2100
2850
2350
2650
P. sandbergii
Live
NS
1550
1500
1900
NS
NS
NS
NS
Dead
NS
NS
NS
NS
2350
NS
NS
NS
A. longiseta
Live
NS
NS
NS
NS
1850
2100
2350
2400
Dead
800
NS
NS
NS
NS
1700
1800
2100
A. mill
Live
1350
1300
2750
3600
3725
3900
4400
5325
efol ium
Dead
1600
NS
NS
NS
5750
5250
6367
4750
A. frigida
Live
2650
3300
3400
-
5150
5450
4250
5550
Dead
2725
NS
NS
NS
NS
NS
4200
3850
-------
APPENDIX A
TABLE A-8. ZAPS II - TREATMENT D (HIGH)
1976 COLLECTION
Sulfur Levels (ppm)
Col lection
Date
9 April
13 May
31 May
21 June
7 July
3 August
15 August
2 September
Jul ian
Date
100
134
152
173
189
216
228
246
A. smithii
Live
1400
1850
3250
4350
4200
4150
3100
3950
Dead
450
NS
NS
NS
NS
5675
3950
4450
K. cristata
Live
1625
1300
1550
3750
4183
3550
3550
3950
Dead
600
NS
NS
NS
NS
3250
3800
2900
P. sandbergii
Live
NS
1400
1900
2400
NS
NS
NS
NS
Dead
NS
NS
NS
NS
1800
NS
NS
NS
S. viridula
Live
NS
3250
2150
4650
4850
3600
4150
4250
Dead
500
NS
NS
NS
NS
3000
3800
4000
A. mill
Live
500
2600
2800
4983
3875
4100
4950
6000
e f o 1 i urn
Dead
550
NS
NS
NS
4750
5400
5500
4550
A. frigida
Live
1450
2700
3550
-
4850
5150
5450
5100
Dead
850
NS
NS
NS
NS
NS
5925
4350
-------
APPENDIX B
TABLE B-l. COLLECTION OF Agropyron smithii AT TWO ANACONDA MONITORING
SITES, 1976.
Site
Sulfur Concentrations (ppm)
Highway Junction
Monitoring Site
3500
3550
4025
3750
3700
3000
Average 3587
Mill Creek
Monitoring Site
2400
2150
1950
1950
2050
1950
2650
2950
3000
1950
Average 2300
Arithmetic Average 0.02
Maximum 1 hr. Ave. 1.52
Maximum 3 hr. Ave. 1.25
Maximum 24 hr. Ave. 0.48
Number of Monitoring Days 329
Total 1 hr. Readings Taken 7665
Arithmetic Average 0.014
Maximum 1 hr. Ave. 0.79
Maximum 3 hr. Ave. 0.48
Maximum 24 hr. Ave. 0.13
Number of Monitoring Days 317
Total 1 hr.Readings Taken 7202
TABLE B-2. PPM SULFUR IN Agropyron smithii FROM ZAPS I, TREATMENT D
collected February 16, 1977.
1450
1550
1300
1350
1100
1500
1250
1200
1450
1500
x = 1365
= 149
s- = 47
x
n = 10
471
-------
APPENDIX B
UJ
CO
CM
O
CO
s
a.
a.
UJ
to
o
h-
o
10-
0
AT 0,10 PPM
FORiHR. READINGS
CONTINUOUSLY
AT 0.05 PPM
CONTINUOUSLY
AT 0.02 PPM
CONTINUOUSLY
HWY JCT(I976)
MlLLCR.(l976
AT 0.005 PPM CONTINUOUSLY
HWY JCT
MILL CR
ZAP ID (1975)
ZAP I D
ZAP 1C (1975)
ZAP IB (1975)
ZAP 1 A (1975)
_j
UJ
3600 >
o:
u_ —
-J x
-27OOS S|
co10
-I 2
< O
&>
-1800.H- o
2 Q;
o Q
2 <
Ul UJ
-900 ?Q
L
u.
o
o
10
I I I • •
APR. MAY JUNE JULY AUG.
SEPT. OCT.
Figure B. Dosage and accumulation of sulfur in Agropyron smithli.
-------
SECTION 14
INVESTIGATION OF THE IMPACT OF COAL-FIRED POWER PLANT
EMISSIONS UPON INSECTS: ENTOMOLOGICAL STUDIES AT THE
ZONAL AIR POLLUTION SYSTEM
by
J.J. Bromenshenk
INTRODUCTION
This portion of the report concentrates on the entomological investiga-
tions at the EPA sulfur dioxide fumigation plots (Zonal Air Pollution
System-ZAPS) at Taylor Creek, near Fort Howes, Montana, for the periods of
June 15 through October 20, 1975, and May 1 through November 15, 1976.
During 1975, attention was given primarily to collecting baseline data in
the vicinity of Colstrip, Montana, since baseline conditions would no longer
prevail there once the Colstrip power plants began operation. The first
plant began to burn coal in early September 1975. With baseline data banks
established (see sections on animal and vegetation studies in the vicinity
of Colstrip, Montana, by Gordon, Tourangeau, and Rice and the entomological
investigations by Bromenshenk), attention was redirected to the ZAPS.
Investigations were initiated in the late summer of 1975 and efforts
were intensified in 1976. The reason for the intensification of the work at
ZAPS was twoTold: (1) The baseline data collections were completed and the
study had entered a monitoring phase; and (2) we were convinced that the
Zonal Air Pollution System could be useful in testing the effects of sulfur
dioxide (S02) upon plant and animal (arthropod) species or populations.
This belief was based on first season evaluations of field experiments, the
system design, and testing of the gas delivery. These indicated that meas-
urable changes in response to the S02 had occurred (unpublished reports to
EPA).
Experiments were conducted to study the behavioral and physiological
responses of honeybees to jjn situ S02 fumigation in 1975, and in 1976,
studies were initiated of the inter- and intra-plot distributions of other
insect species: Ground-dwelling beetles, airborne or flying insects, and
native pollinators.
As noted previously, this research is one aspect of the Colstrip
project to develop methods to predict the bioenvironmental impacts of
emissions from coal-fired power plants before any damage actually occurs.
The most prevalent toxic, gaseous materials released by the combustion of
473
-------
coal are sulfur oxides (DHES, 1975; WES, 1973). Because of their abundance,
consequences to human and vegetative health, and deteriorative effects,
oxides of sulfur are significant air pollutants (Smith, 1975). They may
impact ecosystems as gaseous oxides, as acids in precipitation (Likens,
1974), or as synergists with other pollutants.
Known interactions of insects and sulfur compounds include: (1) The
use of S02 as a fumigant against stored product and household pests (Kanaga,
1956; Swisher, 1944; Negherbon, 1966); (2) changes in the population dynam-
ics of forest insects induced by S02 air pollution stress (Linzon, 1966;
Bosener, 1969; Donaubauer, 1968; Scheffer and Hedgcock, 1955; Kudela, 1962;
Sierpinski, 1966, 1967; Templin, 1962); (3) decreases in populations of
social bees and parasitic wasps and increases in aphids linked to S02
stress from a 615 MW coal-fired power plant in Pennsylvania (Hillmann and
Benton, 1972); (4) decreases in populations of ground-dwelling beetles,
mostly Carabids, near a Kraft mill in Canada (Frietag et al_. , 1973), and (5)
reductions in brood rearing, reduced pollen collection, depressed flight
activity, and uncertain mortality effects produced by fumigation for 9- and
14- week periods using 0 to 5 ppm S02 on 20 honeybee colonies (Hillmann and
Benton, 1972). They noted that reduced flight activity contributed to
decreased honey production.
These literature reports suggest: (1) Trees or plants which are
physiologically weakened by air pollutants such as S02 become more suscep-
tible to attack by those insects which seek out weak plants for feeding and
reproduction; (2) host-predator and host-parasite imbalances may occur,
probably as a result of a concentration of toxic substances in food-chains,
and (3) pollinators, namely honeybees and social bees, demonstrate a variety
of physiological and behavioral responses to S02. In his review of air*
pollution and apiculture (1972), Debackere discussed the chemical reactions
in the atmosphere which convert S02 to sulfurous acid (H2S03) and sulfuric
acid (H2S04). He reported that it is well known that these sulfur compounds
have an irritating effect on the gastrointestinal tract of mammals. Although
almost nothing is known about the effects on bees as indicated by literature
reports, it is assumed that these acids have a deleterious effect on the
intestinal tract of bees. The ground beetle studies of Frietag et al.
(1973) demonstrated significant decreases in the numbers of Carabids associ-
ated with increased fall-out of S042 in the form of sodium sulfate (Na2S04);
these beetles were all predatory species. Kulman (1974) Deported that
Carabids are sensitive to most insecticides and pollutants and that it
appeared that seed-eating beetles contain fewer ppm of insecticides than
predatory beetles. A bibliography of literature pertaining to air pollution
and insects is included as Appendix A in section 5; a more in-depth litera-
ture review appears in the section on the entomological studies near Col-
strip.
In the introduction to a problem analysis of air pollution in urban
areas which divides air pollution influences on temperate forest systems
into three classes based on pollution load, Smith (1975) presented the known
responses of trees to these loads and indicated associated ecosystem impacts.
This breakdown provided a generalized conceptual model for other ecosystems;
his model is presented in Table 14.1.
474
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TABLE 14.1. INFLUENCE OF AIR POLLUTION ON TEMPERATE FOREST ECOSYSTEMS1
Designation
Air Pollution
Load
Response of Trees
Impact on Ecosystems
Class I
Low
Class II
Intermediate
en
Class III
High
1
2.
1 .
Act as a sink
contaminants
for
No or minimal physiological
alteration
Reduced growth
a. detruded nutrient
avail abi 1 ity
1) depressed litter
decomposition
2) acid rain leaching
b. suppressed photosynthesis
enhanced respiration
Reduced reproduction
a. pollinator interference
b. abnormal pollen, flower,
seed, or seedling
development
3. Increased morbidity
a. predisposition to
entomological or
microbial stress
b. direct disease induction
1 . Acute morbidity
2. Mortality
1. Pollutants shifted from
atmospheric to organic or
available nutrient com-
partment
2.
Undetectable influence or
fertilizing effect
Reduced productivity,
lessened biomass
Alteration of
composition
species
Increased insect outbreaks,
microbial epidemics
1. Simplification: increased
erodibility, nutrient
attrition, altered micro-
climate and hydrology
2. Reduced stability
Smith, 1975
-------
Because the Colorado investigative team (CNREL) conducted tests to
determine the effects of sulfur dioxide fumigation on the density, biomass,
and trophic composition of above-ground and below-ground arthropods, this
research concentrated on a few insect species which, according to literature
reports, were sensitive or susceptible to perturbations, easy to monitor and
measure, and reliable predictors of bionomic and economic impacts. In
addition, these species could be related to the studies in the Colstrip area
and to the needs of the overall project. Honeybees were used as detectors
and accumulators of air pollutants at apiary locations throughout south-
eastern Montana. These bees have an estimated economic value of $6 to $33
million as pollinators and $300 to $725,000 as honey producers annually
(Montana Agricultural Reports, 1977; McGregor, 1976), have bionomic value as
pollinators of rangelands and croplands, and have proved to be extremely
sensitive and useful indicators of a variety of atmospheric pollutants (see
section on Colstrip vicinity studies). Smith (1975) listed pollinator
interference and reduced reproduction, which alters plant species composi-
tion, as an important effect of intermediate air pollution stress on eco-
systems. Therefore, high priority was placed on studying the effects of S02
fumigation on honeybees.
Data collected by the Colorado team in 1975 (unpublished reports)
indicated that during the first year of fumigation no order of arthropods
showed any treatment response except the Lepidopterans, which are plant
tissue and nectar feeders. They were lowest in the highest fumigation (10
pphm) plot; however, low population numbers made the results inconclusive.
The Colorado investigators noted that scavengers, represented primarily by
Collembolan species, decreased in numbers in the high plot and that numbers
of Carabid beetles also may have changed in response to the highest fumi-
gation levels. Again, the population numbers were too low to make any
definitive statement. Finally, more grasshopper eggs were found on the
control plot, an area that from observations had less cover and somewhat
different soil characteristics than the other plots. The Colorado group
added investigations of grasshoppers in 1976; the investigators of this
research added studies of ground-dwelling beetles (predators and scavengers)
and of native nectar feeders (Lepidopterans, and pollinating Dipterans and
Hymenopterans, namely native bees).
The main objectives of the entomological studies at ZAPS were: (1) To
establish honeybee colonies for behavioral and physiological exgerimentation;
(2) to conduct preliminary tests of experimental procedures and equipment to
maximize the potential usefulness of the ZAPS system; (3) to evaluate the
initial responses of honeybees and other insects to j_n situ S02 fumigation
so that further research directions (e.g., process and mechanism studies)
could be determined, and (4) to evaluate intra- and inter-plot distribution
of the selected species of insects via grid trapping and gradient analysis.
We were interested in characterizing species of insects and temporal
(season) differences in susceptabi1ity to S02 stress in the cool season
grassland. The hypotheses were that: (1) Insects of the higher trophic
levels such as predators, scavengers, and pollinators would be more suscep-
tible to damage than foliage feeders; (2) the effects of S02 fumigation
would be reflected by behavioral and physiological responses of the target
476
-------
organism; (3) these behavioral and physiological responses could be observed
and measured (e.g., distributions, immigration, emigration, mortality,
shortened life span, changes in fecundity and fertility, etc.); and (4) that
long-term chronic exposure to S02 would be reflected by increased suscepti-
bility, increased resistance, elimination, or no discernible effects.
MATERIALS AND METHODS
To reduce physiological and/or behavioral responses due to genetic
variability, in May 1975, the USDA Bee Stock Center (Baton Rouge, Louisiana)
provided eight dihybrid sister queens which had been artificially insemina-
ted with sperm from drones of a single inbred line (see Appendix B for a
description). These queens were introduced into eight colonies and used to
convert the colonies to the "same" genetic composition. Four colonies were
moved to the Taylor Creek site; the other four were left at Broadus, Montana,
to serve as controls and to provide replacements. By late August 1975, all
of the original queens had died or disappeared, although daughter queens
continued three colonies at Taylor Creek and one colony at Broadus. Dr.
Harbo of the Bee Stock Center believed that the problem of the supercedure
of the queens was due to inadequate sperm reserves and/or to injury caused
during artificial insemination. The Bee Stock Center provided replacements
in May 1976. This line of bees has continued in use because they have
proved to be docile in temperament and as such caused a minimum of disturb-
ance to investigators at the ZAPS sites. Due to the lag time of reestab-
lishing colonies from introduced queens, four colonies of "local" bees,
provided by a commercial keeper, were utilized at ZAPS I in 1976,
The glass observation hives described in the Second Interim Report were
not colonized in 1975 because of the supercedure problem.
During the 1975 and 1976 growing seasons, a beehive was set 70 m above
(north) each of the ZAPS I treatment plots. The hives were not set on the
plots because of potential encounters between bees and the crews conducting
the biological sampling. It was requested that the gas delivery line be
extended from each of the treatment plots to the respective hive, but the
EPA design engineers concluded that this might alter the delivery flow
patterns of the entire system. Thus, we could perform only those experi-
ments which involved the foraging of the bees on the plots, but could not
control the exposure, if any, of the bees to the S02.
Because of uncertainty about the dosages of S02 to which the honeybees
were being exposed and because we were interested in the effects of S02 on
native pollinators, a trapping series was begun in 1976 designed to capture
samples of several species of solitary and social bees (Hymenoptera) and
flies (Dipterans) on each of the treatment plots. Drop-traps were based on
a design by Olsen, Elliot, and Associates (1976) which was economical,
practical, and well-suited for this purpose. They reported that these traps
appear to act as artificial flowers. Each trap was composed of a 14-ounce
disposable white, plastic beverage container. A white, plastic, conical
disposable coffee cup from which the bottom had been removed was inserted
into the mouth of each beverage cup. The coffee cup was held in place by
477
-------
a piece of strapping tape. Thus, each trap was a simple funnel-trap. Each
cup was set into the ground so that the upper two-thirds extended above the
surface of the ground. Approximately one inch of water was placed in the
bottom of each beverage cup to drown any captured insects. The cups were
inspected and insects removed at approximately ten-day intervals. Twenty of
these traps were installed on each of the eight treatment plots for a total
of 160 traps (Figure 14.1).
The data from the CNREL group suggested that ground-dwelling beetles
were affected by the S02; literature reports suggested the same. Therefore,
a network (Figure 14.1) of pit-fall traps was set on each treatment plot.
The traps consisted of the nested cup design utilized to obtain pollinators.
Olsen, Elliott, and Associates (1976) found that sinking these cups deeper
into the ground decreased their attractiveness to pollinators but greatly
increased the catch of beetles (personal communication, D. Schmidt, ento-
mologist). In each treatment plot, 24 of these traps were arranged into a
grid pattern and sunk into the ground so that the mouth was level with the
surface of the soil. In addition, ten extra cups were placed on ZAPS II
below the S02 gas delivery line of the high (10 pphm) concentration fumiga-
tion plot in areas of severe vegetation burn, and ten extra cups were placed
on the control plot in the same relative position as the cups on the high
concentration plot. Thus, each network consisted of 212 collection points
(Figure 14.1) utilized for approximately ten-day intervals during the first
half of each month from May through September. An inch of water in the
bottom of each trap drowned the beetles and prevented the predacious species
from devouring each other and the other captured insects.
The number of beetles obtained in this manner declined rapidly after
mid-June and introduced a problem of random-baiting effect because a trap
containing a few beetles became much more attractive than one without any
beetles. To minimize this effect and to increase trapping success, all
traps were baited with decayed meat. This appeared to resolve the immediate
problem and greatly improved the trapping yield of Scarabaeids and Silphids
(Figure 14.2).
We recognize the limitations of pit-fall trapping and of the possible
sources of error resultant from baiting. However, we agree with the state-
ment of Kulman in his 1974 review of the ecology of the North American
Carabidae in which he remarked that the pit-fall method persists in studies
of ground-dwelling beetles because "...other sampling methods (mark capture,
total population over unit area, etc.) also have problems and are more
difficult to use."
Finally, to study the distribution of pollinators and of other airborne
or flying insects over the plots, which the D-Vac sampling systems utilized
by the Colorado entomological investigative team did not adequately sample,
a series of cylindrical, sticky traps were employed. Each trap consisted of
a 14-ounce disposable plastic beverage cup, coated with Tack Trap® and
taped above each junction of the delivery pipeline (Figure 14.2). The
sticky traps were replaced every two weeks. Since color may affect capture,
both white and yellow cups were used. Reportedly, native pollinators are
attracted to white (Olson, Elliott, and Associates, 1976), while aphids
478
-------
73m
5 •
4 •
T /
0 3 •
L
2
•
• 15 •
•-— A " '•
o
•
•
7
*
— z
8 •
\
c
14 •
• (
i3~~"'
• 24"
• ft
1~A
D
'
' 0-S
17
^_
o
>.— °
(
«MH«B^
12
^
o — /:
18"
D
/
i
2~«
\ —
22?
^—
— ^
21
V.
19 *•
o
m •
\ — i
•
\ *«HB
•
1
"
10
20
Mixing shed
A Sticky-traps
• Pit-traps
• Water-traps
o Pit-traps in vegetation burn areas
Figure 14.1. Distribution of insect traps on ZAPS plots - Summer 1976
479
-------
Figure 14.2.
A. Sticky-trap^tapBd to S02 delivery line. B. Coating
sticky-trap cup. C. Pit-fall trap and captured beetles.
Cup was raised slightly for photographic purposes.
480
-------
are attacted to yellow (Broadbent, 1948). Hi 11 man reported increased aphid
numbers in areas subjected to elevated levels of sulfur from a Pennsylvania
coal-fired power plant.
Presumably, color has the greatest effect at low wind speeds (Southwood,
1975). Taylor (1962) demonstrated that a white sticky trap caught insects
almost as if they were inert particles, and he constructed tables relating
the efficiency of cylindrical traps for different insects at different wind
speeds. The relation of catch to size of sticky traps was investigated by
Heathcote (1957) and by Staples and Allington (1959). They found that catch
usually increased with trap size, but this increase was not proportional to
trap size; the smallest trap caught the largest number of insects per unit
area. Based on these findings, we decided to test the effects of color
During each six-week trap period, ten white and ten yellow stock-cups were
used on each treatment plot for two weeks, then 20 yellow cups for two
weeks, followed by 20 white cups for two weeks, at the end of which time the
cycle was repeated. When two colors were used, the colors were alternated
in each row of traps.
Captured insects were frozen until they could be identified and counted.
The number of insects captured per cup per trap day was calculated for
purpose of analyses. Destroyed (hail, mice, wind) or upset cups were not
included in the analyses. All traps (sticky, drop, and pit-fall) were laid
out in a grid system based on the pattern established by the gas delivery
line. The pipe junctions were used as sampling points. Each cross-shaped
junction visually marked out four quadrants. Quadrants falling outside the
north and south perimeter of the delivery system were considered to fall
outside of the treatment plot. The traps were arranged in a systematic
manner so that each junction and quadrant was utilized, each quadrant being
replicated an equal number of times within a plot. Trap locations from
treatment plot to plot were identical. Within each "junction quadrant" the
pit-fall or drop trap location was determined by measuring 125 cm from the
horizontal gas pipeline diagonally to the ground at the first orifice away
from the junction. This placed each trap just over one meter from the line.
A systematic grid trapping network was used to study dispersion patterns as
well as changes in population size.
During 1975, dead-bee traps, pollen traps, magnetic labels, and feeding
stations were used in tests on honeybee responses to the fumigations. The
dead-bee traps consisted of plastic traps below the entrance of each hive,
enclosed by a 2-cm mesh to the inch screen. Living bees could easily fly or
crawl through the screen, but housecleaning bees could not carry the dead
bees out. Dead bees in the traps were counted daily during those periods
when investigators were at the ZAPS plots. Numbers of bees that died away
from the hive were not included in the analyses.
The original design for a pollen trap, which is part of the dead bee
trap (EPA, 1976), was based on a design by Hillmann (1972). However, this
design restricted the entrance to the hive, and the bees piled up at the
entrance o'n days when they were actively foraging. Therefore, a pollen trap
based on the design by Henderson (1962) was used. This consisted of a deep
honey super (box) with a 2-cm mesh screen over its top, which then became
481
-------
the new entrance. The pollen trap was set under the rest of the hive struc-
ture. Bees entered via a normal entrance, then crawled through the screen
which formed a false bottom in the hive. The screen brushed pollen off the
bees' legs. A pull-out drawer lined with plastic sheeting allowed for the
removal of the pollen.
We used a ferrous metal tag capture-recapture technique (Gary, 1971)
when appropriate for distribution, movement, and flight tests. Coded metal
tags were glued to the abdomen of bees captured in the field. These tags
were retrieved at the entrance to the hives by a strip of ceramic magnets.
The magnets either pulled the tags off the bees as they entered, or else
both tags and bees stuck to the magnet but the bees were able to wiggle free
within a short period leaving the tag behind. The ferrous metal technique
demonstrated a 78 percent retrieval success, which is consistent with the
success ratio reported by Gary (1971). Gary discussed reasons for less than
100 percent retrieval in his paper.
For some experiments, bees were marked with dots of enamel paint
(color-coded to each hive). Direct counts of bee visitations and foraging
on each plot or at feeding stations were made by walking transects across
the plots.
Behavioral tests using feeding stations (glass-watch glasses on which
jars were inverted and filled with a one molar sucrose solution as described
by von Frisch, 1967) were conducted in late August and early September of
1975. At that time very few flowering plants were in blossom, and honeybees
were easily and rapidly trained to feeding stations. During this same 1976
period, honeybees were attracted to the water-containing drop traps and
captured, although at other times honeybees were seen only infrequently in
or around these traps. Apparently, the shortage of water in nearby reser-
voirs forced the bees to the traps.
Chi square analyses of variance were used to test whether number of
honeybees or other insects on the fumigation plots fit an equal distribution
ratio or whether numbers fit some other ratio such as dependency on fumiga-
tion level. Regression and co-variance analyses were applied where appro-
priate, but for the 1976 period the tests relied on the theoretical levels
of mean S02 concentration because the actual data were not available. These
analyses are useful approximations but the actual data are needed before
more intensive and accurate mathematical methods can be applied, such as
within plot distribution.
RESULTS
Chi square analyses of variance were used to test whether numbers of
honeybees on the fumigation plots fit an equal distribution ratio or whether
numbers of bees were dependent on the concentration of S02- The following
observations were made at the ZAPS sites during 1975:
482
-------
(1) Honeybees fed at all feeding stations, even when a feeding station
was placed directly under a point source emitting concentrated S02.
(2) The number of honeybees at each feeding station increased from the
time each experiment was initiated, indicating either recruitment or dis-
covery by other bees.
(3) The number of bees at a feeder was greatest just after the sucrose
solution was depleted and then decreased rapidly if the feeder was not re-
filled.
(4) The bees generally were not successful in locating a feeding
station that was hidden from sight, even when fragrance (an essential oil
such as peppermint) was added.
(5) Marked and tagged bees from each of the colonies (located 75 m
north of the fumigation areas) were observed on the plots; no preferential
response to S02 as indicated by the number of bees visiting the feeding
stations or different plots could be discerned (Table 14.2).
TABLE 14.2. VISITS BY ADULT WORKER HONEYBEES TO FEEDING DISHES
(ONE MOLAR SUCROSE), ZAPS I, AUGUST-SEPTEMBER, 1975.
Experiments 1-6. Total Number of Bees/Plot Versus Fumigation Level
(Theoretical Levels)
Number of Bees
CONTROL LOW MEDIUM HIGH X2 P
91 118 127 109 6.38 >0.05
Experiments 3 and 4. Total Number of Bees/Plot at Dishes a) Hidden in Grass
with No Fragrance; b) Hidden with Essential Oil Fragrance; c) Not Hidden,
e.g., white 20 cm squares under dishes. (30 minute tests).
CONTROL
LOW
MEDIUM
HIGH
Number of Bees
a)
b)
c)
0
1
172
0
3
124
0
0
169
0
0
120
During both 1975 and 1976, mortality of the bees at the ZAPS site
appeared to be normal; no obvious lethal effects could be discerned. It
should be noted that the hives were not placed directly on the fumigated
plots primarily because of concern expressed about possible problems asso-
ciated with honeybees and researchers in close proximity.
The bees did not appear to avoid different concentrations of S02 gas
nor did they exhibit obvious signs of intoxication, either lethal or sub-
lethal, such as interference with foraging activity. But in 1975, the bees
did not successfully locate feeding stations hidden from view, even if a
fragrance was added. Odor may be more important than vision as a guidance
system to bees, and the addition of small amounts of an essential oil should
483
-------
be a strong attractant (von Frisch, 1967). It is possible that the S02
interfered with the bees' olfactory sensory systems or that other odors are
more attractive than peppermint.
The long-term effects of continued exposure to S02 are still not known,
but it appeared that during the summers of 1975 and 1976, a large portion of
the bees from each colony did not forage on the control or fumigated plots
and as such, may have had little contact with the S02.
Acetylcholinesterase (Ache) enzyme analyses on heads or brains from
bees (both .foraging and dying bees), which were presented in the portion of
this report describing the chemical analyses of bee tissue from sites
throughout southeastern Montana, were performed by R. Barker, Bee Research
Laboratory, Tucson, Arizona. He found that the rates of enzyme activity of
bees from the ZAPS I site and controls from apiaries near Broadus, Montana,
were as high or higher than the normal activity rates of bees from the
Tucson area and considerably higher than the rates of enzyme-activity in
bees poisoned by organo-phosphate pesticides. Failure to detect an S02-
induced decrease in the Ache activity of bees at the ZAPS sites may be due
to avoidance of the S0? since the hives were not placed on the treatment
plots and the bees were free to forage elsewhere. Foraging outside of the
ZAPS plots may have been related to a lack of good forage materials on the
plots rather than avoidance of the sulfur, which from the feeding tests
appeared to be the case.
During 1976, the queens from each hive, except the one above the LOW
plot, superceded, leaving daughter queens to take their places. However,
the hive above the MEDIUM plot was left in a queenless state. This colony
was replaced in late June with a colony from the Broadus area and an extra
colony was set above the control plot to use in case of additional super-
cedures. Because of the supercedures, in the fall the strength of the
colonies and the amount of stored honey ranked from the largest to smallest
was: LOW, CONTROL, MEDIUM, HIGH.
In late September of 1976, honeybees began to seek out the drop-traps
for water, as indicated by their lapping of water from the sides of the
trap. The result was that bees were captured in these traps at this time of
the year when they had not been captured before. For example, by walking a
transect across the plots, the numbers of bees on the plots near these traps
(September 21, 1976) was 31, 25, 2, and 2 for the CONTROL, LOW, MEDIUM, and
HIGH treatments, respectively. However, because of the supercedures, these
numbers approximated the difference in the strength and size of the colonies
nearest each plot.
Bees from the ZAPS I site in 1975 had slightly lower sulfur content (x
= 4,000, SD = 436) than those obtained in the previous year near Col strip
(x = 4,392, SD = 286). In 1976, bees from ZAPS I had lower sulfur (x =
2,508, SD = 504) in late June than in August (x = 3,990, SD = 942). However,
in August, the bees near Colstrip contained more sulfur (x = 4,933, SD =
506) than bees from ZAPS. Hillmann (1972) found lower sulfur in larval bees
than in adult bees. In June, bee colonies would have contained a greater
proportion of young bees than the same colonies in August.
484
-------
Currently, the drop-trap captures are being processed, and those data
will be presented in the next report. The traps worked well and were very
specific to the collection of bees and flies. The pollen they were carrying
verified them as pollinators.
We also are in the process of completing the sticky trap evaluations.
So far, we have not found any demonstrable trends in numbers across the
plots. Yellow sulfur butterflies (Colias eurytheme Bdv.) were attracted to
yellow sticky traps over white traps. The numbers were significantly
different for some plots (Table 14.3) but did not correlate to the S02
treatment levels.
The deposition of small insects on the sticky traps may be useful for
the interpretation of prevailing wind patterns on the plots at the level of
the pipeline, assuming that the insects are deposited more or less as inert
particles as suggested by Taylor (1962). Wind roses, based on insect depo-
sition for each plot, are presented in Appendix C. This information has
been included because the instrumentated wind data are not yet available
from EPA, and because there is only a single wind monitor at each of the
ZAPS sites, and it is located approximately three meters above the surround-
ing terrain and may not produce readings representative of wind patterns at
the level of the pipes. The insects counted on the traps for this purpose
were small, weak fliers such as aphids and thrips, which would most likely
be active during the daylight hours.
To date, the identifications and counts of beetles in the pit-fall
traps have been completed for about 80 percent of the traps. Figure 14.3
presents linear regression lines (based on theoretical fumigation means)
which depict the population changes of beetles of the genus Canthon (mostly
Canthon laevis), a decomposer dependent on fecal and dead organic materials
(Ritcher, 1958). The trapping data (Appendix D) for May at ZAPS I versus
ZAPS II shows that more beetles were present on each of the ZAPS II plots.
The number of beetles captured on ZAPS I in May was low compared to ZAPS II,
but the population numbers increased until they were only slightly lower
than those at ZAPS II in August. Also, the number of beetles captured on
the LOW plot at ZAPS I in May was unusually low (21) compared to other plots
and dates, which produced a positive regression slope; but the number of the
beetles on the CONTROL plot exceeded those at any other ZAPS I plot. Com-
puting a regression line for the number of beetles on the CONTROL, MEDIUM
and HIGH plots gave a negative slope, which is still less steep than that
for ZAPS II in May.
The relatively low, beetle populations on ZAPS I in May could be due to
a residual effect from the previous year's fumigations, to S02 levels above
the desired delivery concentrations on some of the plots during the spring
equipment adjustments, to grazing influences (ZAPS II was fenced in 1976 and
ZAPS I in 1975) or to attraction of beetles to rodents caught in traps which
had inadvertantly been left on ZAPS I during the first part of May.
485
-------
TABLE 14.3. STICKY-TRAP CAPTURE OF COLZAS EURYTHEME (SULFUR BUTTERFLIES)
FOR 1520 TRAP DAYS, FIVE SAMPLE PERIODS, N = 985.
Period Ending
Mean number
Total Capture
SD
SE
Goodness of Fit
Total Capture
Plot by Plot
ZAPS I
S02
ZAPS II
S02
CONTROL LOW MEDIUM HIGH CONTROL LOW MEDIUM HIGH
July
Aug.
Aug.
Sept.
Sept.
19, 1976
5, 1976
16, 1976
2, 1976
19, 1976
35
50
18
10
14
18
55
26
9
3
28
27
29
7
12
24
35
22
19
10
26
26
43
19
27
28
25
44
13
13
21
41
29
10
8
54
36
38
13
20
25.4 22.2 20.6 20.0
16.7 20.3 10.3 9.0
7.5 9.1 4.6 4.0
X2 = 2.7, df = 3, P >0.4
28.2 24.6 21.8 32.2
8.9 12.8 13.7 16.1
4.0 5.7 6.1 7.2
X2n = 9.6, df = 3, P <0.05
X2n = 109.9, df = 1, P <0.001 X2n = 5.0, df = 1, P <0.05
for HIGH vs MEDIUM
X2n = 5.1, df = 1, P <0.05
for MEDIUM vs CONTROL
for HIGH vs LOW
X2n = 1.3, df = 1, P >0.3
for CONTROL vs HIGH
By August, the Canthon beetle populations on the control plots of both
ZAPS I and II were very similar, and the regression slopes describing the
population decreases across the plots at both sites were almost identical,
indicating that the populations of this beetle were influenced by the in-
creasing S02 concentrations. We have not presented significance levels for
regression because at the time of this report data concerning the measured
levels of S02 in the ambient air were not available.
The results of chi square analyses to test significance of the differ-
ent total catches for each plot were highly significant (Appendix D). This
conclusion is consistent with that of Frietag et al_. (1973), who reported a
negative correlation between the numbers of ground beetles (primarily preda-
tory Carabids and one carrion-feeding Silphid) and sulfate fallout from a
Kraft mill.
486
-------
700 -r
600 --
500 ..
oo
tt 400 --
55
CO
200
TOO
MAY 1976 (NOT BAITED)
AUGUST 1976 (BAITED)
ZAP I
CONTROL LOW MEDIUM
SULFUR DIOXIDE
HIGH
Figure 14. 3.
Number of beetles (Canthon sp.) in pit-fall traps/S02 treat-
ment plot, plotted against theoretical fumigation levels.
In May, significant differences were documented (X2 analysis) (Appendix
E) at both sites in the numbers of five species of beetles representing
three families (Carabids, Silphids, and Scarabaeids). In most cases, the
number of beetles tended to decline sharply (Figure 14.4) as the S02 con-
centration increased. Later the capture numbers of all but the Canthon
beetles and of the Silphids (Nicrophorus spp.) decreased to very low levels
on all plots, including the controls, and the numbers of Silphids no longer
showed any demonstrable relation to S02 level (X2 analyses).
487
-------
00
00
oo
>-
Q
Q-
T
<
Q
I 30-
o
o
ro
to
!- 20-
111 t-u
LiJ
CO
U_
O
CQ
s:
ID
10-
•Trox sonorae
v Pasimachus elongatus
CONTROL
MEDIUM
S02 CONCENTRATIONS
HIGH
Figure 14.4A. Pit-fall captures of beetles on ZAPS I treatment plots, May 1976.
-------
-P=>
00
540 -
oo
-------
DISCUSSION
The honeybee work at ZAPS I has been limited by the need to place the
hives off the treatment plots. They may receive some drift from the plots,
but this has not been measured. Extension of the delivery lines to the
hives appears to be impractical. Moving of the hives onto the plots would
insure exposure but could result in interference with the sampling by other
investigators, at least one of whom is known to be allergic to bee stings.
Bees forage on the plots; one colony swarmed onto the HIGH plot in June 1976
and settled under a small shelter from which they had to be removed. A
greater number of bees in the drop traps in September 1976 on the CONTROL
and LOW pphm plots more likely was related to the fact that there were
smaller, weaker colonies above the higher plots than to an avoidance of the
sulfur, since in 1975 the bees showed no differential avoidance.
The supercedure rate seems rather high and exceeds that of similar
colonies in Broadus. In 1975, we thought this problem was a consequence of
artificial insemination. In 1976, queens were used that had been insemi-
nated during a nuptial flight, and were selected to insure healthy colonies
and queens. Since the queen may live for several years, while workers only
live for a few weeks or months at the longest, the queen is usually the
oldest member of the colony and as such would be exposed to any environ-
mental contaminants for a longer period than the workers. The inability to
detect significant differences in total sulfur between bees at the ZAPS
plots and bees in other areas may be due to a masking effect from the high
natural levels of sulfur of animal tissues bound in proteins. Hillmann
(1972) reasoned that this explained the lack of significant increases in the
sulfur content of adult worker honeybees fumigated with as much as 5 pphm of
S02 for 9 and 14 weeks as compared to non-fumigated controls. He found that
S02 produced significant reductions in brood rearing which led to correlated
declines in pollen collection. Both brood rearing and pollen collection
showed an inverse linear relationship to increasing S02 concentrations.
Hillmann thought that reduced brood production may reflect an effect of S02
on the queens. Also, he found that control colonies exhibited significantly
more flight activity than fumigated colonies but saw no significant differ-
ences in the degree of flight suppression among the three fumigation treat-
ments. Reduced flight activity contributed to lessened honey and colony
weight. The toxic effect of S02 in Hillmann's studies was unclear: fumi-
gated bees showed significant mortality increases in the firs£ year of the
study but not in the second year. He did not observe behavioral abnormali-
ties induced by S02 fumigation based on his observations of bees in a glass-
observation hive. Acute dosages of approximately 1,000 ppm S02 indicated
that adults were more sensitive than larvae, eggs, and pupae, respectively.
However, we believe that it is unlikely that these extreme levels of S02
would be encountered in the ambient air
Available and preferred forage is often better off of the ZAPS plots
than on because there are nearby alfalfa and clover hay fields. Thus, it is
likely, as observations suggested, that the majority of the bees in each
colony did not forage the ZAPS treatment areas, including the control,
unless something attracted them onto the plots such as the feeding stations
or the water in the drop-traps.
490
-------
Since significant increases in fluorides in bees were found near Col-
strip and a probable toxic effect was identified at one site (see Colstrip
vicinity studies, this report), it is believed that honeybees are particu-
larily efficient and sensitive bioindicators of some major and trace ele-
ments. Sulfur and sulfur compound impacts may be more difficult to identify
and measure, but Hillmann's data (1972) indicate that sulfur dioxide does
have significant impacts on honeybees. In 1977, we intend to move the bees
into areas where the bees may have a greater probability of exposure to S02,
such as in the buffer zones, and the levels to which they are exposed using
sulfation plates will be monitored at each hive. Behavioral responses such
as flight activity and physiological responses such as mortality and brood
rearing influenced by S02 will be examined. For comparison purposes, tech-
niques will be similar to those used by Hillmann.
Hillmann (1972) found that tissues of larval or immature bees had
significantly less sulfur than those of adult bees. The bees at the ZAPS
plots contained less sulfur in June than in August. June is a period of
rapid population increase; whereas August is a period of a relatively stable
or declining population size. Thus, the bee populations in June would have
had a greater proportion of young adult bees, which probably accounted for
the lower sulfur content. Bees at the ZAPS plots consistently demonstrated
lower sulfur levels than bees at the other sites. Also, there seemed to be
a tendency towards lower sulfur in both bees and beetles on the HIGH treat-
ment plot. Lower sulfur content in insects on plots fumigated with S02
could occur as a consequence of rapid population turnover or shortened life
span. This would increase the proportion of young to old members of the
population. Assuming that sulfur increases in older insects, this could
explain the observed trend.
Data from the beetle trapping are important as regards soil cycles.
The Scarabaeidae and the Silphidae gather, bury, and break down organic
material. This in turn releases nutrients to the soil systems. There are
several literature reports that support the hypothesis that air pollutants,
particularly heavy metals, accumulate in litter and the upper soil horizons
and interfere with nutrient cycling. In a review on depressed litter decom-
position and reduced nutrient availability, Smith (1975) stressed the need
for studies on the effects of contaminants on these processes and their
ultimate effect on plant growth. Also, many contaminants are toxic to
microorganisms, particularly fungi, which are primarily responsible for the
decomposition of organic materials in the soil (Horsfall, 1956). A number
of insects, including the dung beetles such as Canthon, disperse spores of
saprophagic or coprophilous fungi (Lodha, 1974). These insects act both as
agents of mechanical breakdown and movement of organic materials and as
disseminators of primary microbial decomposers. Thus, any reduction in the
populations of these beetles could have effects on the decomposition and
nutrient cycles.
Based on the data, the numbers of Canthon beetles on ZAPS I built up
through the summer to the level of the beetles on the ZAPS II plots. This
indicates that what may be occurring is a depressed immigration or popula-
tion growth on the fumigated plots rather than an elimination or migration
491
-------
of the beetles from these plots. Waldow (1973) discovered an antennal cell,
a coelosphaerica, in carrion beetles (Nicrophorus vespilloides) which
responds to the odor of carrion, H2S, and a few cyclic compounds. This
receptor may be responsible for a coding of the odor of rotten meat. Sulfur
dioxide could act in a somewhat similar manner, interacting on the insects'
ability to orient to food material by olfaction. This can be tested in a
simple y-tube olfactometer patterned on the design by Burkholder (1970).
Data from the other sampling procedures at ZAPS as well as within-plot
distributions of insects, will be presented in the next interim report.
SUMMARY AND CONCLUSIONS
We have focused our studies on ground-dwelling beetles and on honey-
bees—insects which our studies and those of the Colorado investigators
indicate are most susceptible to S02.
Both the types of experiments and the results of experiments using
honeybees at the ZAPS plots were limited by the placement of the beehives 75
m above each of the treatment plots. This averted possible problems for
investigators from bees in close proximity. However, it introduced a new
problem since only bees foraging on the plots would be expected to receive a
full strength exposure of short duration to the S02 gas. The bees did not
demonstrate measurable avoidance of the sulfur gas on any of the plots, nor
did they exhibit obvious signs of sublethal effects such as intoxication or
of lethal effects as indicated by mortality. However, bees did not success-
fully locate feeding stations hidden from view, even if an attractive fra-
grance was added to the sugar solution. Odor may be more important than
vision as a guidance system to bees, and the addition of small amounts of
essential oils should be a strong attractant (von Frisch, 1967). It is
possible that the S02 interfered with the bees' olfactory sensory systems or
that odors other than those tested are more attractive.
In May, several species of ground-dwelling beetles, mostly Scarabaeids
and Silphids (carrion beetles), demonstrated significant inverse relation-
ships in numbers compared to S02 fumigation concentration. By late summer,
tumblebugs (Canthon spp.) continued to demonstrate this relationship. The
response of carrion feeders was variable, and the capture of other beetle
species was too low for meaningful comparisons. The inverse 'relationships
of numbers to S02 levels appeared to be related to movement of beetles onto
the plots, since the beetle numbers on the ZAPS I plots increased during the
season. The increase was greatest on the control and the low fumigation
treatments. Hydrogen sulfide apparently is an attractant to carrion beetles
(Waldow, 1973). Since hydrogen sulfide attracts carrion beetles, it was
hypothesized that other sulfur compounds may act in a similar manner, and
the release of S02 gas over large areas could mask the ability of beetles to
detect and to orient towards food sources of decaying organic materials.
The sulfur content in honeybees reared at the ZAPS plots during 1975
and 1976 tended to be somewhat lower than the sulfur content of bees at
sites in other areas of southeastern Montana. Also, the sulfur content of
492
-------
bees at ZAPS was relatively low in June and increased by August 1976. In
both bees and beetles, there appeared to be a trend towards decreased sulfur
in insects on the HIGH treatment plots. Since Hillmann (1972) reported that
immature bees had significantly less sulfur than older bees, it was suggested
that decreased sulfur in insects exposed to S02 could be explained by high
turnovers or shortened life spans of the members of a population, which
would result in a higher proportion of younger to older individuals.
The data processing and analyses of insects other than beetles captured
by the trapping systems (pit-fall, drop-trap, and sticky-trap) in 1976 were
not complete at the time of this report, but this information should be
available for the next interim report.
Emphasis on sampling programs (population estimates and population
dispersal) will be continued but will be restricted to selected species and
refined to maximize trapping or capture precision and success, applicability
to statistical analyses, and efficiency in terms of man-hours. Trapping
regimens will be limited to three or four periods coinciding with seasonal
population cycles, replacing more or less continuous sampling at considerable
effort and expense. The intent is to de-emphasize repetitive sampling and
redirect efforts toward process- and mechanism-oriented experiments to
determine the basis for observed responses to S02.
At present, beetles seem to offer the greatest potential for studies at
the ZAPS plots. Not only do they appear to respond to S02 but they include
important predators, saprophages, and necrophages. The latter two groups
undoubtedly play a role in litter decomposition and nutrient cycling.
Rigorous investigations of one or two major species of beetles and of the
major ecological events and processes interacting with these populations are
needed. Also, the role of these species in the grassland ecosystem and soil
ecosystem must be appraised. This requires both quantitative and qualitative
information regarding behavior, seasonal cycles, and population dynamics
relative to -changes in biotic and abiotic components of the ecosystems in
order to evaluate the chronic effects of S02 stress. This work would
contribute both to the goals of the overall project as regards the impact of
S02 and would provide valuable information concerning soil systems and the
ecology of soil animals. It is particularly well-suited to the current
needs of the project in view of a lack of information on soil systems and of
the cessation of above-ground arthropod sampling and re-directed emphasis by
the Colorado investigators on soil arthropods.
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Broadbent, L. 1948. Aphis Migration and the Efficiency of the Trapping
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493
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Burkholder, W. E. 1970. Pheromone Research with Stored-Product Coleoptera.
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Debackere, M. 1972. Industriele luchtvervuiling en bijenteelt. (Industrial
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DHES (Montana Department of Health and Environmental Sciences), Bureau of
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Donaubauer, E. 1968. Sekundarschaden in Osterreichischen Rauchschadens-
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Kanaga, E. E. 1956. An Evaluation of the Use of Sulfur Dioxide in Fumigant
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496
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APPENDIX A
See Appendix A in the section 5 of this report by Bromenshenk entitled:
INVESTIGATION OF THE IMPACT OF COAL-FIRED POWER PLANT
EMISSIONS UPON INSECTS: ENTOMOLOGICAL STUDIES IN THE
VICINITY OF COLSTRIP, MONTANA
497
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APPENDIX B
DESCRIPTIONS OF THE INBRED LINES OF HONEYBEES
UTILIZED IN THE ARTIFICIALLY INSEMINATED
QUEENS OBTAINED FROM THE USDA BEE STOCK CENTER.
Queens were a dihybrid YDGk.
Drones were a Pa line.
YD is a yellow line. Queens are stocky. Brood rearing is initiated early
in the season and continues late into the fall. They winter poorly, con-
suming lots of honey and often producing inadequate stores.
Gk is an intermediate color between black and yellow. The workers are
small. The queens do not lay eggs well. Honey production is low. Win-
tering is poor. Workers are marked by 1 or 2 yellow bands.
YDGk hybrids produce a good line.
^a is a yellow line. The queens are large and are the best layers of the 18
lines at the Bee Stock Center. Brood rearing begins early and ends late in
the season. Population size is good during the foraging season. The bees
store little honey and are very susceptible to American Foul Brood.
YDGkxPa makes a good cross.
498
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APPENDIX C
Wind roses derived from the deposition of wind-carried insects on
sticky traps. The length of each line indicates the relative frequency of
insects captured and as such the prevailing wind pattern during the daylight
hours. The angle indicates wind direction relative to the layout of the
plots, thus a horizontal line indicates wind current perpendicular to the N-
S axis of the plots. In all cases the lines indicate wind directed towards
the point of intersection.
499
-------
ZAPS I WIND ROSES, JULY 15, 1976
Control Plot
Low Plot
\
Medium Plot
High Plot
en
O
O
Figure C-l
Control Plot
ZAPS I WIND ROSES, AUGUST 5, 1976
Low Plot
Medium Plot
High Plot
Figure C-2.
-------
ZAPS I WIND ROSES, AUGUST 16, 1976
Control Plot
Low Plot
Medium Plot
Hiqh Plot
en
o
Figure C-3.
ZAPS I WIND ROSES, SEPTEMBER 2, 1976
Control Plot
Low Plot
Medium Plot
High Plot
Figure C-4.
-------
ZAPS I WIND ROSES, SEPTEMBER 19, 1976
Control Plot
Low Plot Medium Plot
Figure C-5.
High Plot
en
o
ro
-------
ZAPS II WIND ROSES, JULY 15, 1976
en
O
CO
Control Plot
Control Plot
Low Plot Medium Plot
Figure C-6
ZAPS II WIND ROSES, AUGUST 5, 1976
Low Plot Medium Plot
Figure C-7.
High Plot
High Plot
-------
Control Plot
en
O
ZAPS II WIND ROSES, AUGUST 16, 1976
Low Plot Medium Plot
Figure C-8.
ZAPS II WIND ROSES, SEPTEMBER 2, 1976
Control Plot
Low Plot
Medium Plot
High Plot
High Plot
Figure C-9.
-------
Control Plot
ZAPS II WIND ROSES, SEPTEMBER 19, 1976
Low Plot Medium Plot
Figure C-10.
High Plot
en
O
en
-------
APPENDIX D
NUMBER OF BEETLES (CANTHON SP=) IN PIT-FALL TRAPS/S02 TREATMENT PLOT.
May 1976--300 trap days/piot--not baited
Site Number of Beetles Computations
ZAPS I CONTROL 240 Y = 141.908 + 1.198X
LOW 21 r = .0565
MEDIUM 149 without LOW data
HIGH 148 Y = 220.000 - 16.200X
All plots 558 r = .667
X2 = 174.2, df=3, P<0.005
ZAPS II CONTROL 589
LOW 374 Y = 527.216 - 35.426X
MEDIUM 358 r = .932
HIGH 185
All plots 1506
X2 = 218.3, df=3, P<0.005
August 1976 - 120 trap days/plot - Baited
Site Number of Beetles Computations
ZAPS I CONTROL 767
LOW 476 Y = 658.405 - 44.154
MEDIUM 371 r = .894
HIGH 269
All plots 1883
X2 = 294.1, df=3, P<0.005
ZAPS II CONTROL 864
LOW 443 Y = 683.934 - 43.573X
MEDIUM 352 r = .764
HIGH 336
All plots 1995
X2 = 370.0, df=3, P<0.005
9
August 1976 - 50 trap days/plot - Baited
(Vegetation Burn Areas)
CONTROL 426
HIGH 66
X2 = 263.4, df=3, P<0.005
^Regression computations are based on theoretical rather than measured S02
delivery concentrations.
506
-------
APPENDIX E
X2 EVALUATION (EXPECTED VALUES EQUAL) BEETLE CAPTURES
ON ZAPS I AND II TREATMENT PLOTS IN MAY 1976.
Species
Canthon sp.
Nicrophorus sp.
Qnthophagus sp.
Trox sp.
Pasimachus sp.
Canthon sp.
Nicrophorus sp.
Qnthophagus sp.
Trox sp.
Pasimachus sp.
CONTROL
589
153
136
43
39
240
38
315
12
8
LOW
374
100
131
19
13
21
9
22
0
5
Pic
MEDIUM
ZAPS I
358
26
92
12
20
ZAPS II
149
32
189
17
7
)t
HIGH
185
21
49
6
11
178
20
109
1
8
X2
218.3
160.4
48.1
39.5
23.6
173.4
20.15
292.9
27.9
.86
P
P<0.005
P<0.005
P<0.005
P<0.005
P<0.005
P<0.005
P<0.005
P<0.005
P<0.005
P<0.5
507
-------
SECTION 15
THE RELATIVE SENSITIVITY OF SELECTED PLANT SPECIES
TO SEVERAL POLLUTANTS SINGLY AND IN COMBINATION
by
D. T. Tingey, L. Bard, and R. W. Field
INTRODUCTION
With the introduction of coal-fired power plants into grassland eco-
systems it becomes important to recognize and determine the possible effects of
power plant emissions on the ecosystems. There have been only limited studies
on native prairie plant species either describing injury symptoms or providing
relative sensitivity data (Davis et al_. , 1966; Hill et ah , 1974).
The objectives of this study were to describe the air pollution-induced
symptoms and to determine the relative sensitivity of selected plant species to
common air pollutants, singly and in combination.
METHODS AND MATERIALS
PLANT GROWTH
Plant growth and exposures to air pollutants were conducted in greenhouse
facilities in Corvallis, Oregon. The plant material used in the study (Table
15.1) was collected in the Otter Creek Valley, Montana, near the Fort Howes
Ranger Station during August, 1974. Since then, the individual species were
propagated vegetatively; the grasses by division and Fringed Sage Wort by
cuttings. The plants were grown in 225 ml styrofoam cups containing a 2:1
(v:v) mixture of perlite:Jiffy Mix. Plants for all studies were grown in
greenhouses; watered daily with North Carolina State University phytotron
nutrient solution (Downs and Hellmers, 1975) and periodically leached with
water. The plants were grown at day/night temperatures of 26-32/18^0; the
sunlight was supplemented and the light period extended to 16 hr/day with light
from HID sodium vapor lamps. Maximum light intensities were 450-650
microeinstenins M-2 sec-1.
PLANT EXPOSURES
Pollutant exposures were conducted in single pass exposure chambers
located in a greenhouse (Heck, Dunning, and Johnson, 1968). Sulfur dioxide and
nitrogen dioxide diluted in nitrogen were metered into the exposure chambers at
a rate sufficient to maintain the desired gas phase concentrations. Sulfur
508
-------
TABLE 15.1. PERCENT FOLIAR INJURY OF PLANTS EXPOSED TO SULFUR DIOXIDE FOR 4
HOURS1
Plant Species
0.5
Agropyron smith ii Rydb
WESTERN WHEAT GRASS 0
Bouteloua gracilis Lag.
BLUE GRAMMA 0
Koeleria cristata (L.) Pers.
PRAIRIE JUNE GRASS 0
Stipa comata Trin. and Rupr.
NEEDLE & THREAD GRASS 0
Artemisia frigida Willd.
FRINGED SAGE WORT 0
Triticum aestivium L. em Thell.
WHEAT, HYSLOP 0
S02 Cone.
1.0
0
3
0
0
0
!
0
(ppm)
1.5 2.0
0 3
3 6
1 6
0 2
6 13
4 5
1 Plants were exposed for 4 hr and foliar injury was assessed 96 hr after
exposure. The injury was measured as the percentage of the leaf area
showing S02 injury. Each mean was the average of 9 observations:
Sx=1-
TABLE 15.2. PERCENT FOLIAR INJURY OF PLANTS EXPOSED TO NITROGEN DIOXIDE FOR
4 HR1
„
Plant Species
WESTERN WHEAT GRASS
BLUE GRAMMA
PRAIRIE JUNE GRASS
NEEDLE & THREAD GRASS
FRINGED SAGE WORT
WHEAT, HYSLOP
0.5
0
0
0
0
0
0
N02 Cone.
1.0
0
3
0
0
0
0
(ppm)
2.0
1
4
1
1
0
0
4.0
1
8
4
1
0
1
1 Plants were exposed for 4 hr and foliar injury was assessed 96 hr after
exposure. The injury was measured as the percentage of the leaf area
showing N02 injury. Each mean was the average of 9 observations:
509
-------
dioxide was measured with a flame photometric sulfur dioxide analyzer; nitrogen
dioxide and ozone were monitored with chemiluminescence analyzers. Prior to
each exposure the analyzers were calibrated using known concentrations of each
pollutant. The plants were exposed to the pollutant 4 to 6 weeks after
vegetative propogation. Plant injury was visually assessed in 5% increments 96
hours following exposure as the percentage of the plant leaf area exhibiting
visual pollutant effects.
RESULTS
PLANT INJURY DESCRIPTIONS
Plant injury resulting from sulfur dioxide, nitrogen dioxide and ozone was
generally similar to previous reports (Jacobson and Hill, 1970). Injury from
sulfur dioxide and nitrogen dioxide was similar on the monocots. On young
leaves, injury developed at the leaf tip, on older leaves injury usually
occurred at the bend of the leaf. Injury usually occurred as small bifacial
lesions, and as severity increased, the lesions coalesced and spread up and
down the leaf. Interveinal streaks of necrotic tissue were frequent. Lesion
color ranged from light tan to ivory. On Fringed Sage Wort the injury occurred
on the middleaged leaf tissue as a bifacial collapse of the tissue, killing
both veins and intervenial areas. The ozone-induced injury on Fringed Sage
Wort was similar to that caused by sulfur dioxide. Ozone injury to monocots
appeared as tip necrosis and small necrotic lesions between the veins; as the
interveinal lesions coalesced, necrotic streaking resulted. Injury tended to
be concentrated between the leaf tip and bend in the leaf. Injury that
resulted from the mixtures of sulfur dioxide + nitrogen dioxide or sulfur
dioxide + ozone were similar to sulfur dioxide injury.
RELATIVE SENSITIVITY
To determine the relative sensitivity of each of the plant species, they
were exposed to either single pollutants or mixtures of pollutants and their
injury rated. Wheat was included in all studies as an example of an agri-
cultural crop. All species were injured following a four-hour exposure to 2
ppm S02, with Fringe Sage Wort exhibiting the most injury and Needle and Thread
Grass the least (Table 15.1). Only Blue Gramma was injured at a concentration
of less than 1.5 ppm.
In exposures to nitrogen dioxide (Table 15.2) only Prairie June Grass and
Blue Gramma developed significant injury with Blue Gramma being more sensitive.
All plants except Needle and Thread Grass were injured by the lowest ozone
concentration (0.4 ppm) (Table 15.3). Blue Gramma and wheat were the most
sensitive, Western Wheat and Needle and Thread Grass the least sensitive.
Plants were exposed to either mixtures of sulfur dioxide + nitrogen
dioxide (Table 15.4) or sulfur dioxide + ozone (Table 15.5) to determine if the
gas mixtures interacted to alter the injury response of the plants. Injury was
very similar on plants exposed to sulfur dioxide with or without nitrogen
dioxide. When plants were exposed to sulfur dioxide with or without ozone,
510
-------
TABLE 15.3. PERCENT FOLIAR INJURY OF PLANTS EXPOSED TO OZONE FOR 4 HR1-
Plant Species
WESTERN WHEAT GRASS
BLUE GRAMMA
PRAIRIE JUNE GRASS
NEEDLE & THREAD GRASS
FRINGED SAGE WORT
WHEAT, HYSLOP
0.4
1
6
5
0
2
9
03
0.6
2
12
5
0
4
11
Cone, (ppm)
0.8
5
15
11
2
11
17
0.95
7
25
12
2
12
23
1 Plants were exposed for 4 hr and foliar injury was assessed 96 hr after
exposure. The injury was measured as the percentage of the leaf area
showing 03 injury. Each mean was the average of 9 observations, S- = 2.
TABLE 15.4. PERCENT FOLIAR INJURY OR PLANTS EXPOSED TO SULFUR DIOXIDE OR
SULFUR DIOXIDE PLUS NITROGEN DIOXIDE1.
Plant Species S02 Cone, (ppm) S02 Cone, (ppm) + 0.1 ppm N02
073 O O T72 O 076O T772~~
WESTERN WHEAT GRASS
BLUE GRAMMA
PRAIRIE JUNE GRASS
NEEDLE & THREAD GRASS
FRINGED SAGE WORT
WHEAT, HYSLOP
0
0
0
0
0
0
0
1
0
0
0
0
2
3
1
0
1
1
4
4
3
0
4
1
0
0
0
0
0
0
3
1
1
0
0
0
4
3
1
0
0
1
5
4
2
0
0
1
Plants were exposed for 4 hr and foliar injury was assessed 96 hr after
exposure. The injury was measured as the percentage of the leaf area
showing injury. Each mean was the average of 8 observations: S- = 2.
511
-------
injury was similar except in Fringed Sage Wort where the gas mixture caused
less injury than the sulfur dioxide alone.
TABLE 15.5. PERCENT FOLIAR INJURY OF PLANTS EXPOSED TO SULFUR DIOXIDE OR
SULFUR DIOXIDE + OZONE1.
Plant Species
S02 Cone. (PPM)
075O ^
SO
1.5 2.0
I2 Cone. (PPM) + 0.1 ppm 03
7T5 O T75 27o~
WESTERN WHEAT GRASS
BLUE GRAMMA
PRAIRIE JUNE GRASS
NEEDLE & THREAD GRASS
FRINGED SAGE WORT
WHEAT, HYSLOP
0
1
0
0
0
0
1
2
1
0
9
4
3
2
3
1
24
4
3
2
4
2
33
6
2
1
0
0
0
1
2
3
0
0
0
1
3
2
2
0
4
2
4
4
3
0
6
3
Plants were exposed for 4 hr and foliar injury was assessed 96 hr after
exposure. The injury was measured as the percentage of the leaf area
showing injury. Each mean was the average of 8 observations:
DISCUSSION
Sx=1'
The injury observed on the native species was similar to that previously
described (Jacobson and Hill, 1970; Hill et al_. , 1974). Injury thresholds
suggested for the species tested in these experiments do not take into account
the effect of soil water potential on plant sensitivity. Low soil water poten-
tial could reduce plant sensitivity. The levels of pollutants used in this
study are higher than would usually be expected in the field. They were used
only to provide concentrations that would readily cause visual injury to aid in
injury description and relative sensitivity ranking.
Davis et ah (1966) reported that levels of sulfur dioxide that might be
expected around a Phelps Dodge smelter in Arizona, and which would defoliate
cocklebur, did not injure Blue Gramma. Hill et al_. (1974), working with
established native plants in the field, showed that Agropyron caniun and A.
desertorum required between 6 and 10 ppm S02 for two hours to cause visual
injury. This suggests that these two species of Wheat Grass are more tolerant
of S02 than Western Wheat Grass. Hill watered the plants for a mtinth prior to
exposure to ensure that they were in a stage of rapid growth and thus highly
sensitive. He also reported that Stipa occidental is required 10 ppm S02 for
induce visual injury. This suggests that Stipa occidental is is
than Stipa comata which showed injury at 2 ppm S02 for four
two hours to
more tolerant
hours.
Previous reports (Tingey et al_. , 1971; Tingey et al_. , 1973) of green-
house studies suggested that mixtures of sulfur dioxide + nitrogen dioxide or
sulfur dioxide + ozone interacted to cause more foliar injury than the effects
of the single gases. These observations were not substantiated in this study.
The gas mixtures did not increase the injury over that caused by sulfur dioxide
alone. The lack of synergism from sulfur dioxide + nitrogen has previously
512
-------
been reported for native plants (Hill et al., 1974) and in growth chamber
studies (Bennet et al., 1975).
REFERENCES
Bennet, J. H., A. C. Hill, A. Soleimani and W. H. Edwards. 1975. Acute effects
of combination of sulphur dioxide and nitrogen dioxide on plants.
Environmental Pollution 9:127-132.
Davis, C. R. , D. R. Howell and G. W. Morgan. 1966. Sulphur dioxide fumigations
of range grasses native to southeastern Arizona. Journal of Range
Management 19:60-64.
Downs, R. J. and H. Hellmers. 1975. Environment and Experimental Control of
Plant Growth. Academic Press. New York, New York.
Heck, W. W. , J. A. Dunning and H. Johnson. 1968. Design of a simple plant
exposure chamber. DHEW, National Center for Air Pollution Control,
Publication APTD-68-6. Cincinnati, OH. 24pp.
Hill, A. C. , S. Hill, C. Lamb and T. W. Barrett. 1974. Sensitivity of native
desert vegetation to S02 and to S02 and N02 combined. Journal Air
Pollution Control Association 24:153-157.
Jacobson, J. S. and A. C. Hill. 1970. Recognition of Air Pollution Injury to
Vegetation. A Pictorial Atlas. Air Pollution Control Association.
Pittsburg, PA.
Tingey; D. T. , R. A. Reinert, J. A. Dunning and W. W. Heck. 1971. Vegetation
injury from the interaction of nitrogen dioxide and sulfur dioxide.
Phytopathology 61:1506-1511.
Tingey, D.. T. , R. A. Reinert, J. A. Dunning and W. W. Heck. 1973. Foliar
injury responses of eleven plant species to ozone/sulphur dioxide
mixtures. Atmospheric Environment 7:201-208.
513
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SECTION 16
PROGRESS IN MODELING THE EFFECTS OF S02 FUMIGATION
ON AN EASTERN MONTANA GRASSLAND
by
J. L. Dodd, M. Coughenour, and W. K. Lauenroth
INTRODUCTION
The modeling effort is charged with (1) adapting ELM (the grassland
ecosystem simulation model) to the Montana sites; (2) developing and implem-
enting simulated direct effects of atmospheric S02 on ecosystem components; and
(3) developing a sulfur submodel to examine the sulfur cycle. Work is
proceeding ahead of schedule in the cases of (2) and (3) and is slightly behind
schedule in the case of (1).
ADAPTATION OF ELM TO LOCAL SITES
Adaptation of ELM to the mixed-grass prairie ecosystem has proceeded
through completion of the application and validation of the abiotic submodel.
Initial conditions, parameter values, and validation information for the
producer, decomposer, nitrogen, and phosphorus submodels are completed.
The investigations are currently examining procedures for model
adaptation. Information developed by the Natural Resources Ecology Laboratory
(NREL) modeling team and supported by new findings of Dave Tingey (personal
communication) suggest that modification of the structure of the producer
submodel will lead to considerable conceptual strengthening of the whole model,
especially in regard to the effects of S02 on the ecosystem. The dilemma,
which is influenced by limited funding, is whether to proceed wit|j adaptation
of the currently implemented model and then make the new changes in that
version, or wait a few months until the new and more conceptually sound version
of ELM is available and then modify that version. The best representation of
the effects of S02 will be incorporated into the new version. In either case,
the data-based information needed for adaptation will be essentially the same
as that now being completed.
DEVELOPMENT AND IMPLEMENTATION OF SIMULATED DIRECT EFFECTS OF ATMOSPHERIC S02
Currently the NREL Grassland Biome staff is modifying the structure of the
primary producer submodel to more accurately represent the biomass dynamics of
grassland ecosystems. A current draft of the new structure is shown in Figure
16.1. The most important changes to be incorporated will be (a) the
514
-------
Old
Standing Dead
Recent
Standing Dead
en
en
Photosynthetic
Pool
Mature
Leaves
Juvenile
Leaves
Annual Stems
and Flowers
Perennial Stems
Live Roots
Figure 16.1, Structure of the new version of the primary production submodel now being implemented by the
NREL modeling staff. The significant changes in the structure are the addition of an
explicit photosynthetic pool compartment and the splitting of the existing leaf compartment
into two compartments, mature and juvenile leaves.
-------
photosynthetic production of a pool of translocatable carbohydrates and (b) the
partitioning of this photosynthate among various portions of the plant. The
investigators feel that the new treatment of these two processes in plant pro-
duction will be especially well suited for showing the direct effects on plants
of inorganic sulfur taken up as sulfur dioxide. It is hypothesized that a
close interrelationship exists between the process of photosynthesis and
partitioning of photosynthate. Field experiments for summer of 1976 are being
formulated to test these hypotheses.
A new feature of modeling mature leaves separate from juvenile leaves will
also be amenable for showing the effects of sulfur taken up as sulfur.dioxide,
since these two leaf categories are expected to exhibit differential effects.
Related to this concept is the phenology of the plant and the varying effects
with the age of the plant. The plant phenology model will also be improved
before the final coupling to the sulfur cycling mode.
Other effects on primary producers are expected to be shown on the
processes of (a) transpiration, as reflected by changes in stomatal resistance,
(b) uptake of sulfate from the soil as reflected by changes in the sulfur
status of the plant, and (c) plant death, as reflected by chlorosis and
necrosis.
If significant differences are shown to exist among major species of the
mixed-grass prairie with regard to sulfur dioxide uptake rates and related
effects on the above-mentioned processes, the primary producer model will be
able to reflect these changes as altered community composition. Modeled
functional groups would be selected with regard to similarities of fundamental
production processes, and similarity of responses to sulfur dioxide.
DEVELOPMENT OF SULFUR SUBMODEL TO EXAMINE SULFUR CYCLE
The direct effects of sulfur dioxide pollution on primary production will
depend not only on the amount of sulfur dioxide present in the atmosphere, but
also on its distribution throughout the plant system, and the closely related
soil system. A sulfur cycling model is ideally suited to demonstrate these
relationships, and a current draft of its structure is shown in Figure 16.2.
The state variables and processes are selected to maintain a level of resolu-
tion comparible with ELM, and similarity exists with the existing nitrogen and
phosphorus models. Not only will the effects of pollution be shown on primary
producers, but also on the entire sulfur cycling system, a capability set in a
more entire ecology perspective.
Atmospheric sulfur inputs will be in the form of sulfur dioxide, and the
acid-rain sulfate. Rainfall inputs will be simply a function of rainfall,
allowing coupling to atmospheric models which facilitate prediction of
concentrations in rainfall. Gaseous inputs to plants might be described by the
fol lowing scheme:
V - 1 1 1 F
516
-------
01
Primary Producer
Af Abiotic
'-"\
(BIO) Soil Blotlc
Figure 16.2. Structure of the sulfur submodel for ELM now being developed to represent the dynamics of
sulfur cycling in grassland ecosystems.
-------
Where V~ = deposition velocity
(j
r = stomatal resistance
r = atmospheric resistance
a
r. . = internal plant resistance
int. p
F = flux into plant
C(Z) = gas concentration at height Z
Where F = atmospheric flux
cl
K(Z) = eddy diffusivity of air at height Z
K(Z) is a function of wind speed and plant height;
r will be a function of light intensity, humidity, soil water, plant
s species, plant condition, temperature;
r. . will be a function of metabolic activity of plant, sulfur
in ' concentration in plant, and plant species.
Gaseous uptake or deposition to the soil and to surface litter will be
treated in analogous manner, with an atmospheric resistance and a surface
resistance. The surface resistance of these two components will be functions
of water content, relative humidity, temperature, and amount of sulfur already
deposited.
Such processes as decomposition of litter may then be altered according to
sulfur concentrations resulting from fluxes. Although the soil pH is expected
to be well buffered in this geographic area, the pH of surface litter may be
altered. This, in conjunction with the altered C:N:S ratio of the litter, may
then affect decomposition. The complexities of the soil sulfur transformations
are demonstrated in Figures 16.3 and 16.4. Although the resolution of the
model will not permit a detailed treatment of these relationships, the major
processes could be represented and subsequently be reflected in the cycling as
shown in Figure 16.1. The great predominance of aerobic versus* anaerobic
processes in this particular system will eliminate much of the complexity
associated with Figure 16.3. A detailed soil chemistry model is not within the
scope of this effort, however, the major processes of Figure 16.4 might be
represented in a rather coarse way.
Another major reason for modeling the soil components, as well as plant,
is that the relationship between plants and soils is an established factor
governing plant distribution and abundance. Any alteration in soil properties
or mineral distributions in the soil may then be expected to play a role in
plant competitive relationships and the related community structure. No effort
is expected to model direct toxic effects of sulfur dioxide upon consumers in
this model, only the consumer relationships to the system by way of plant
consumption.
518
-------
Microbe
Death
Aeroebic
Heterotrophic
Aeroebic \
Reduction J
Anaeroebic
Oxidation
Aeroebic
Oxidation
f Anaeroebic
( Reduction
Aeroebic
Oxidation
Figure 16.3.
Conceptual high resolution of sulfur transformations that poten-
tially occur in soils.
519
-------
so2s
Adsorbed
Bases
Metal
co2
°2
N, P
Mineral
i
r
—
—
Organic
S03
•A
$2°*,
H2S
s
Figure 16.4.
Schematic to represent the complexity of chemical reactions that
can occur in soils.
520
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-78-021
2.
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
The Bioenvironmental Impact of a Coal-fired Power Plant
Third Interim Report, Col strip, Montana
December 1977
5. REPORT DATE
February 1978
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Edited by Eric M, Preston and Robert A. Lewis
8. PERFORMING ORGANIZATION REPORT NO
9. PERFORMING ORGANIZATION NAME AND ADDRESS
U. S. EPA
Con/all is Environmental Research Laboratory
Corvallis, Oregon 97330
10. PROGRAM ELEMENT NO.
EHA541/E-AP-77ACV
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
13. TYPE OF REPORT AND PERIOD COVERED
Interim April - Nnvpmhpr 1975
14. SPONSORING AGENCY CODE
Same
EPA/600/02
15. SUPPLEMENTARY NOTES
In this series of reports, the First Interim Report is EPA number
and the Second Interim Report is EPA number EPA-600/3-76-013.
EPA-600/3-76-002
16. ABSTRACT
The EPA has recongnized the need for a rational approach to the incorporation of
ecological impact information into power facility sitinq decisions in the northern
great plains. Research funded by the Col strip, Coal-fired Power Plant Pro.iect is a
first attempt to generate methods to predict the bioenvironmental effects of air
pollution before damage is sustained. Pre-construction documentation of the en-
vironmental characteristics of the grassland ecosystem in the vicinity of Colstrip,
Montana began in the summer of 1974. Since then, key characteristics of the eco-
system have been monitored regularly to detect possible pollution impacts upon plant
and animal community structure.
In the summer of 1975, field stressing experiments were begun to provide the
data necessary to develop dose-response models for SO^ stress on a grassland eco-
system. These experiments involve continuous stressing of one acre grassland plots
with measured doses of SO^ during the growing season (usually Aoril through October),
Results of the 1975 field season's investigations are summarized in this
publication. The six-year project will terminate in 1980 and a final report will
be published after data analyses are complete.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
plant and animal response to pollution
coal-fired power plant
air pollutants
grassland ecosystems
mathematical modeling
remote sensing
micrometeorological investigation
coal-fired power plant
emissions
air quality monitorina
aerosol characterization
51
8. DISTRIBUTION STATEMENT
release to public
19. SECURITY CLASS (This Report)
Unclassified
21. NO. OF PAGES
561
20. SECURITY CLASS (This page)
Unclassified
22. PRICE
EPA Form 2220-1 (Rev. 4-77) PREVIOUS EDITION is OBSOLETE
H U S. GOVERNMENT PRINTING OFFICE' 1978-799-936/85 REGION 10
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