EPA-600/3-77-045
May 1977
Ecological Research Series
Office of Research and
U.S. Environmental Protection Agency
Duluth, Minnesota
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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-600/3-77-045
May 1977
ALGAL NUTRIENT AVAILABILITY AND LIMITATION
IN LAKE ONTARIO DURING IFYGL
Part II. Nitrogen Available in Lake Ontario Tributary
Water Samples and Urban Runoff from Madison, Wisconsin
by
William F. Cowen
Kannikar Sirisinha
and
G. Fred Lee
University of Texas at Dallas
Richardson, Texas 75080
Contract No. R-800537-02
Project Officer
Nelson Thomas
Large Lakes Research Station
Environmental Research Laboratory--Duluth
Grosse lie, MI 48138
ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
DULUTH, MINNESOTA 55804
U.S. Environmental Protection Agency
Region 5, Library (5PL-16)
230 S. Dearborn St-eet, Boom 1670
Chicago, IL 60604
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DISCLAIMER
This report has been reviewed by the Environmental Research
Laboratory-Duluth, U.S. Environmental Protection Agency, and approved
for publication. Approval does not signify that the contents necessarily
reflect the views and policies of the U.S. Environmental Protection
Agency, nor does mention of trade names or commercial products consti-
tute endorsement or recommendation for use.
ii
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FOREWORD
Our nation's freshwaters are vital for all animals and plants,
yet our diverse uses of watei for recreation, food, energy,
transportation, and industry—physically and chemically alter lakes,
rivers, and streams. Such alterations threaten terrestrial organisms,
as well as those living in water. The Environmental Research Laboratory
in Duluth, Minnesota develops methods, conducts laboratory and field
studies, and extrapolates research findings
--to determine how physical and chemical pollution affects
aquatic life
--to assess the effects of ecosystems on pollutants
--to predict effects of pollutants on large lakes through
use of models
--to measure bioaccumulation of pollutants in aquatic
organisms that are consumed by other animals, including
man
This report provides information on the nitrogen availability
in Lake Ontario tributary waters. The information is required in the
understanding of algal nutrient growth in Lake Ontario.
Donald I. Mount, Ph.D.
Di rector
Environmental Research Laboratory
Duluth, Minnesota
iii
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PREFACE
This project was conducted as part of the International Field Year
for Great Lakes Research and consisted of three separate parts, all direc-
ted toward providing information needed to assess the factors limiting algal
growth in Lake Ontario and the amounts of nitrogen and phosphorus in tribu-
taries and drainage which would likely become available in the lake. Part I
is concerned with a comprehensive study of the amounts of phosphorus enter-
ing Lake Ontario from U.S. tributaries which will likely become available in
the lake. Particular attention is given to the particulate and organic
forms of phosphorus in the major U.S. tributaries to the lake. Part II is
concerned with a study of the amounts of available nitrogen entering Lake
Ontario from the U.S. tributaries. Part III is concerned with the factors
limiting algal growth in Lake Ontario and in the major U.S. tributaries.
This report presents Part II of this study. Parts I and III are published
as separate reports by the U.S. Environmental Protection Agency and entitled,
Algal Nutrient Availability and Limitation In Lake Ontario During IFYGL,
with the following subtitles:
Part I: Available Phosphorus in Urban Runoff and Lake Ontario
Tributary Waters
Part III: Algal Nutrient Limitation in Lake Ontario During IFYGL
iv
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ABSTRACT
Samples of water from the Niagara, Genesee, Oswego,
and Black Rivers were collected from March to June, 1973.
The samples were analyzed for nitrogen forms and were
incubated in darkness under aerobic conditions to promote
mineralization of soluble inorganic nitrogen from the
organic nitrogen in the samples. The amounts of ammonia
and nitrate were determined as a function of the time of
incubation. Generally, over 50 percent of total nitrogen
present in these river samples was immediately available
for algal growth or potentially available after minerali-
zation by bacteria. The results were highly variable from
each tributary and no single value could be selected from
the data obtained to describe the availability of total
nitrogen in a given river.
This report was submitted in fulfillment of Contract
No. R-800537-02 under the sponsorship of the Environmental
Protection Agency. Work was completed as of June, 1975.
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CONTENTS
Foreword ill
Preface iv
Abstract v
List of Figures viii
List of Tables xi
Acknowledgments xii
I Introduction 1
II Conclusions 3
III Recommendations 4
IV Literature Review 5
V Methods and Procedures 11
VI Results 22
VII Discussion 51
References 65
Appendices
A. Nitrogen Mineralization of New York River Waters 68
B. Nitrogen Mineralization from Madison, Wisconsin,
Urban Runoff Samples 68
vii
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LIST OF FIGURES
No. Page
1 Main Biological Process Involving Nitrogen 6
2 Lake Ontario and Major Tributaries 12
3 Percent of Total N As Ammonia-N + Nitrate-N 25
During Dark Incubation of Niagara River
Samples
4 Percent of Total N As Ammonia-N + Nitrate-N 29
During Dark Incubation of Genesee River
Samples
5 Percent of Total N As Ammonia-N + Nitrate-N 30
During Dark Incubation of Oswego River
Samples
6 Percent of Total N As Ammonia-N +• Nitrate-N 31
During Dark Incubation of Oswego River
Samples
7 Percent of Total N As Ammonia-N + Nitrate-N 32
During Dark Incubation of Black River Samples
8 Nitrate-N Production from Genesee R (42) 34
Particles in Lake Ontario (10) Water
9 Nitrate-N Production from Oswego R (43) 35
Particles in Lake Ontario (10) Water
10 Nitrate-N Production from Genesee R. (58) 38
Particles in Lake Ontario (96) Water
11 Nitrate-N Production from Oswego R. (59) 39
Particles in Lake Ontario (96) Water
12 The Mineralization of Runoff Water from 40
Whitney Way Station Collected on October
20, 1972, Incubated Under Aerobic Conditions
13 The Mineralization of Runoff Water from 40
Whitney Way Station Collected on December 30,
1972, Incubated Under Aerobic Conditions
viii
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FIGURES (continued)
No. Page
14 The Mineralization of Runoff Water from 41
Whitney Way Station Collected on January
17, 1973, Incubated Under Aerobic Condi-
tions
15 The Mineralization of Runoff Water from 41
Whitney Way Station Collected on March 5,
1973, Incubated Under Aerobic Conditions
16 The Mineralization of Runoff Water from 43
Manitau Way Station Collected on October
20, 1972, Incubated Under Aerobic Condi-
tions
17 The Mineralization of Runoff Water from 43
Manitau Way Station Collected on December
30, 1972, Incubated Under Aerobic Condi-
tions
18 The Mineralization of Runoff Water from 44
Manitau Way Station Collected on January 17,
1973, Incubated Under Aerobic Conditions
19 The Mineralization of Runoff Water from 44
Manitau Way Station Collected on March 5,
1973, Incubated Under Aerobic Conditions
20 The Mineralization of Runoff Water from 45
Water Chemistry Lab Station Collected on
October 20, 1972, Incubated Under Aerobic
Conditions
21 The Mineralization of Runoff Water from 45
Water Chemistry Lab Station Collected on
December 30, 1972, Incubated Under Aerobic
Conditions
22 The Mineralization of Runoff Water from 46
Water Chemistry Lab Station Collected on
March 5, 1973, Incubated Under Aerobic
Conditions
23 The Mineralization of Runoff Water from 46
Water Chemistry Lab Station Collected on
March 5, 1973, Incubated Under Aerobic
Conditions
ix
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FIGURES (continued)
24- The Mineralization of Runoff Water from 47
Stone Ridge Apartment Station Collected
on January 17, 1973, Incubated Under
Aerobic Conditions
25 The Growth Curve of Selenastrum capri- 50
cornutum Using Nitrate as Nitrogen Source
26 Release of Ammonia and Nitrate in Urban 60
Runoff in Madison, Wisconsin, Station A
(Whitney Way)
27 Release of Ammonia and Nitrate in Urban 60
Runoff in Madison, Wisconsin, Station B
(Manitau Way)
28 Release of Ammonia and Nitrate in Urban 60
Runoff in Madison, Wisconsin, Station C
(Water Chemistry Laboratory)
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LIST OF TABLES
No. Page
1 Urban Runoff Sampling Stations in Madison, 14
Wisconsin
2 Initial N Availability 22
3 Distribution of Organic Nitrogen Forms in 24
River Water Samples
4 Mineralization of Total N to Ammonia and 27
Nitrate in 35-50 Days
5 Mineralization of Organic-N to Ammonia and 28
Nitrate in 35-50 Days
6 Nitrogen Mass Balances 33
7 Initial Concentrations of Selected Nitrogen 37
Species in the Samples Used in this
Investigation
8 The Nitrogen Availability from Particles 49
Using Bioassay Test at 10 Days
9 The Average Concentrations of NHU-N, NO--N 57
and Total Kjeldahl-N in Urban Runoff of
Madison Areas
10 Comparison of Nitrogen Availability from 62
Chemical and Bioassay Method
11 Summarization of the Percent N-Availability 64
in Short-Term and Long-Term Studies of
Urban Runoff
xi
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ACKNOWLEDGMENTS
This investigation was conducted primarily at the
University of Wisconsin. In addition, support was given
the investigation by the U.S. Environmental Protection
Agency. Support was given by the Department of Civil and
Environmental Engineering at the University of Wisconsin
and the Institute for Environmental Sciences at the
University- of Texas at Dallas. The authors wish to
acknowledge the assistance of Nancy Halverson in performing
the analysis of some of the samples. Collection of samples
from the Oswego River was performed under the direction of
Richard B. Moore of the Lake Ontario Environmental Lab-
oratory, State University College, Oswego, New York.
Collection of rain gage water was performed by the staff
of the U.S. Environmental Protection Agency Laboratory,
Rochester, New York, under the direction of D. Casey.
Genesee River basin samples were sent to Madison by the
personnel of New York State Department of Environmental
Conservation, Albany, New York, under the supervision of
Patricia Boulton. We wish to acknowledge the assistance
of Nelson Thomas, project officer for the U.S. EPA for his
assistance in sample procurement.
xii
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SECTION I
INTRODUCTION
Nitrogen is recognized as one of the most important
constituents of living matter. It has been known to be a
probable limiting algal nutrient in many waters. The in-
crease in the loads of nitrogen into surface water in the
presence of other nutrients, leads to the eutrophication
process resulting in the excessive growth of algae and
aquatic weeds and also depletion of oxygen in the hypo-
limnion of lakes that undergo thermal stratification.
There are many sources of nutrients that get into receiving
bodies of water,one of them being urban runoff. Current
trend in the U.S. indicates that more and more of the po-
pulation is being concentrated into urban regions. Al-
though considerable amounts of time and money are being
consumed to handle the sanitary wastes from the urban envi-
ronment, little information is available on the significance
of the stormwater runoff from urban areas.
Previous evidence indicates that the dominant form of
nitrogen compound in urban runoff is organic nitrogen.
These nitrogen compounds come from precipitation, dustfall,
street litter and vegetation. They are generally not avail-
able to the algae for growth. However, after entering the
lakes or rivers, the organic nitrogen compounds can be con-
verted into ammonia by biochemical and physico-chemical ac-
tivities. Under aerobic conditions, the ajnmonia form of ni-
trogen will be oxidized to nitrate. Since nitrogen as nitrate
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and ammonia is readily available for algal growth, urban
runoff represents an important source of nitrogen in the
lakes and rivers.
Samples of water from the Niagara, Genesee, Oswego,
and Black Rivers were collected from March to June, 1973.
Also, samples in the Madison, Wisconsin urban runoff were
collected. The samples were analyzed for nitrogen (N)
forms, then were incubated in darkness under aerobic con-
ditions to promote the regeneration of soluble, inorganic-N
from the organic-N in the samples, by the following series
of reactions:
Organic-N -»• Ammonia-N -»- Nitrite-N ->• Nitrate-N
The progress of the nutrient regeneration process was fol-
lowed by means of ammonia and nitrate analyses made during
the incubations. The amount of nitrogen in a form which
would be available for algae was estimated by the sum of
ammonia-N and nitrate-N and was expressed as a percent of
the total N or of the organic-N in the samples.
A limited series of experiments was also performed to
predict the nitrogen mineralization of particulate matter
from river water in Lake Ontario. Particulate matter was
suspended in Lake Ontario water and incubated under the
same conditions used in the nutrient regeneration studies
with whole river samples. The amount of algal-available
nitrogen produced by the particles was estimated by the
nitrate production and was expressed as a percent of the
organic-N in the particulate matter. Algal bioassay tests
were used to determine the fraction of organic nitrogen
available for algal growth.
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SECTION II
CONCLUSIONS
1. Samples of water from the Niagara, Genesee, Os-
wego, and Black Rivers generally showed over 50 percent of
their total N to be either immediately available for algal
growth or potentially available after mineralization by
bacteria.
2. Because of the variation in total N availability
and small number of samples tested from each tributary, no
single value could be selected from the data to describe
the availability of total N in a given river.
3. Efforts to estimate organic N availability showed
a wide range of values. Low organic N availability values
were found in samples with the high organic N concentrations.
U. Direct estimation of particulate organic-N avail-
ability by dark incubation of membrane-filterable particles
yielded values which conflicted with the availability data
from incubations of the unfiltered river waters.
5. Samples of urban stormwater drainage obtained
from Madison, Wisconsin, showed that over 80 percent of the
total nitrogen in the sample may be converted to an avail-
able form for algal growth in a period of approximately
100 days.
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SECTION III
RECOMMENDATIONS
It is recommended that in those situations where ni-
trogen is thought to be the limiting element controlling
algal growth, studies should be initiated which utilize the
techniques developed in this investigation to determine
what part of the total nitrogen entering the water body
will likely become available to support algal growth in
receiving waters. Additional large-scale studies are need-
ed to determine the factors influencing the results of
available nitrogen from various types of organic and par-
ticulate matter present in natural waters. Particular
emphasis should be given to investigating such factors as
the temperature of the solution, effects of mixing, trace
element composition, and other factors which may influence
the available nitrogen in a particular sample.
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SECTION IV
LITERATURE REVIEW
Mineralization is the process of conversion of or-
ganic matter to inorganic form. This process can be
brought about by changes in solution due to physico-
chemical as well as bacterial action. Bacteria are the
principal agents of conversion of nitrogen from one form
to another in the nitrogen cycle, and the steps of the
cycle of most concern to water quality are those invol-
ving water-soluble nitrogen, namely ammonification, ni-
trification, and denitrification. Decomposition of or-
ganic matter starts with autolysis, which leads to an
increase of permeability, enabling several compounds al-
ready present in dissolved state, to leave the cell. More-
over, compounds in undissolved state may dissolve. The
processes of liberation and mineralization of elements
play an important part in the chemical cycle in fresh water
because it makes nutrients available for use by aquatic
micro-organisms. The main biological processes involving
inorganic nitrogen are shown diagramatically in Figure 1.
Nitrogen in urban runoff is in the forms of nitrite,
nitrate, ammonium and organic nitrogen. Weibel et al_. (1964)
have studied urban land runoff in Cincinnati, Ohio as a fac-
tor in stream pollution and found that most of the nitrogen
is in organic form in the range of 0.2 to 4.8 mgN/1. He
also compared stormwater runoff loads and sanitary sewage
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loads and reported that in stormwater runoff the suspended
loading was 1.4- times that of sewage where COD was 25 per-
cent, BOD 6 percent, phosphorus 9 percent, and nitrogen 11
percent of raw sewage.
Lee et_ al. (1966) and Sonzogni and Lee (1972) had es-
timated the amount of nutrients entering Lake Mendota an-
nually per acre of urban drainage. The results showed that
most of nitrogen in urban runoff was in the organic form as
was found in the Cincinnati study (Weibel et al., 1964).
Sonzogni and Lee also noted that three times more nitrogen
was entering Lake Mendota from urban runoff than the pre-
vious estimation by Lee. It is obvious from this inform-
ation that the urban stormwater runoff cannot be neglected
in considering waste loadings from urban areas.
Several recent reports have suggested that nitrogen
more often might be the limiting factor in aquatic biomass
production than heretofore believed. Gerloff and Skoog
(1957) compared the growth of Microcystis aeruginosa in
sterilized Lake Mendota water enriched with nitrogen, phos-
phorus and other essential elements singly and in various
combinations. The addition of only phosphate or iron pro-
duced no increase in growth while the addition of nitrate
alone resulted in approximately 64 percent of the growth
obtained with all three elements added. They suggested
nitrogen as the important algal growth-limiting nutrient in
eutrophic Lake Mendota. However, conclusions from experi-
ments of this type are limited in their application as the
samples of water represent conditions only in very limited
areas of the lake and at a specific time.
Lund (1965) stated that the low summer nitrate levels
commonly observed in surface water of eutrophic lakes might
suggest nitrogen limitation at that time. Recently, Lueschow
et al. (1970) found indication that the mean monthly nitrogen
-------
(organic or inorganic) and not phosphorus contents of sur-
face waters of Southern Wisconsin lakes provides an index
of their trophic status. This information would suggest
that nitrogen might often be the limiting nutrient in
eutrophic lakes, especially those high in available phos-
phorus .
Golterman (1960) studied the liberation and minerali-
zation of nitrogen during sterile autolysis of Scenedesmus
quadricauda. Autolysis was induced by U.V. irradiation or
by chloroform treatment. Only 20 - 30 percent of nitrogen
compound was liberated. The rest of nitrogen compounds were
in the form of protein and nucleic acids, and remeiined as a
slag. He tried to digest the nitrogen-slag-remairiing by
means of bacteria. When the slag was suspended in lake
water, one half of the nitrogen was converted to ammonia in
5 days, while the other half was divided up into bacterial
and Scenedesmus nitrogen, the mutual ratio of which could
not be determined. This experiment showed that baicteria is
the primary agent responsible for nutrients cycling in water,.
Vaccaro (1965) reported on an experiment in which a
mixed planktonic culture was allowed to decompose. In this ex-
periment, phosphorus regeneration was more rapid than was
nitrogen regeneration. Similar results have been obtained
for nitrogen and phosphorus regeneration from aquatic macro-
phytes (Nichols and Keeney, 1973). Most of nitrogen is in
ammonium form between day 7 to 30 and in nitrate between
day 72 to 80. Temperature and oxygen were found to have
influence in the regeneration process.
Chen, Keeney and Konrad (1972) investigated nitrification
rates in lake sediment-water systems. Sediment samples from
Wisconsin eutrophic and oligotrophic hard and soft-water
lakes were incubated in deionized water at 10 C or 25 C in
the dark in mixed and non-mixed systems, open to the
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atmosphere. The amount of nitrate and ammonium nitrogen were
determined routinely. Nitrate was not formed in the acid
sediments from soft-water lakes. However, in calcareous
sediment from hard-water lakes, nitrification proceeded
readily when the system was stirred to increase oxygen
diffusion. A one to three day lag phase occurred before
nitrification commenced, and in all cases onset of nitri-
fication was accompanied by a corresponding decline in
soluble and exchangeable ammonium nitrogen, the latter
value approaching zero by the end of the incubation period.
Nitrate did not accumulate when the samples were not stirred.
This study indicated that sediments did not add appreciable
amounts of nitrate to water except in a well oxidized, mixed
situation such as might be occurring in shallow areas or
at lake turnover. Keeney (1973) stated that the organic
material in sediments is more stable towards decomposition
than that present in dead plant and animal remains, prob-
ably due to protection by reaction with organic compounds
and clay minerals, formation of resistant heterocyclic
material, and physical inaccessibility. The rapid nitri-
fication rate (270 ygN/1 per hour at 25°C in Lake Mendota),
however, indicated that this process could add appreciable
nitrate to lake waters in turnover time. Temperature also
had effect on nitrification rate, the higher the temperature,
the more rapid nitrification.
Similar results were obtained by Austin (1970). She
studied the release of nitrogenous compounds from lake sedi-
ments. Sediments from hard-water lake (Mendota) released
large concentrations of dissolved inorganic nitrogen under
aerobic conditions. The predominant form of inorganic nitro-
gen leached from the sediment was nitrate. In contrast, am-
monium and soluble Kjeldahl nitrogen were the predominant
species of nitrogen in the anaerobic studies. No detectable
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amounts of ammonia or nitrite were released throughout
the study. The release of nitrogen from hard-water lake
sediment was greater than that of soft-water lake sediment.
She also found that the release of nitrogen from lake
sediment was greatly dependent upon currents and mixing
but not the total nitrogen concentration of a lake
sediment.
Lopez and Galvez (1958) studied the mineralization of
the organic matter to determine the availability of ten
Phillipine soils to release nitrogen in available form under
submerged condition. They found that regardless of the
nature of the soil organic matter and the microflora, the
amount of mineralized nitrogen was significantly and posi-
tively correlated with their contents of organic matter,
total nitrogen, and the nitrogen uptake by rice plants.
Grain yields in greenhouse culture were significantly cor-
related with mineralized nitrogen.
While there have been numerous studies on the conver-
sion of organic and particulate forms of nitrogen present
in soils, lake sediments and algae, there have been no
reported studies on this conversion in urban storm water
drainage.
10
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SECTION V
METHODS AND PROCEDURES
WATER COLLECTION
The Niagara, Genesee, Oswego, and Black Rivers were
sampled during the spring of 1973, at points close to
their discharge into Lake Ontario (Figure 2).
The samples were collected from the 0 to one meter
depth in the rivers and were transported to Madison, Wis-
consin, in plastic containers. No preservatives were ad-
ded to the samples, which were refrigerated at U C upon
reception at the University of Wisconsin Water Chemistry
Laboratory at Madison. The average time between sampling
and initial nitrogen analyses was approximately five days,
of which one to two days were required for (unrefrigerated)
transportation.
URBAN RUNOFF SAMPLES
The main classes of sites selected were; residential
and exposed land under construction. The residential clas-
ses were further divided according to Madison zoning codes
as follows:
R, - A single family residential district, with some
low density multiple family dwellings.
R? - A single family residential district with some
low density multiple family dwellings. This zone differed
from R, in that less usable open space per dwelling unit
was allowed in R_ as compared to R, .
11
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3
.a
.2.
'o*
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C
o
o
C
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-------
Rc - A medium density residential area located in
o
inlying urban parts of the city. The area being sampled
was part of the University of Wisconsin campus.
The urban construction site was an area of Western
Madison undergoing rapid development, with exposed terraces
and severe erosion.
The specific location of each site is given in Table
1, All sites could be sampled within about 45 minutes -
1.5 hours, depending upon traffic conditions. Samples
were collected during rainstorms which appeared to be in-
tense enough for reasonable overland flows. Although
samples taken at various times during a runoff could vary
in concentration of organic nitrogen (Kluesener, 1972),
for qualitative studies of availability, the concentration
was not an important factor. Samples at each site were
taken in one-gallon acid washed plastic cubitainers. All
samples were brought to the laboratory and immediately
stored at 4°C until analyzed.
All chemicals used in these studies were reagent
grade, with the following exceptions: Sodium hypochlorite
was purchased commercially as Clorox (5.25 percent NaCIO),
Selenium oxychloride (SeOCl2> was USP.
EXPERIMENTAL METHODS
Nitrogen in runoff or rivers was considered as soluble
or particles, based on filtration through 0.45 micron
pore-size membrane filters. In this study, available
nitrogen is defined as the nitrogen measured chemically
as nitrate or ammonia or which can be shown to be used
by algae in a growth bioassay. The various nitrogen forms
to be analyzed are:
13
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Table 1. URBAN RUNOFF SAMPLING STATIONS
IN MADISON, WISCONSIN
Drainage Area
Zoning Code
Rl
R2
R5
Urban
Construction
Station
A
B
D
Location
Inlet of the large storm sewer
in the median strip of Whitney.,
near the Montauk place inter-
section.
Inlet of the large storm sewer
in the median strip of Manitau
Way, near the Tumalo Trail
intersection.
Outlet of storm sewer pipe, near
the U.W. Water Chemistry Labo-
ratory, which carries drainage
from the Bascom Hill campus area,
Street gutter flow from the
construction of apartment houses
near the corner of Island Drive
and Masthead Streets, one block
from the Mineral Point Road-
Island Drive intersection.
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Ammonium Nitrogen (NH^-N)
Analysis of a filtered sample, using alkaline phenol
procedure adapted for the Auto Analyzer (Kluesener, 1969).
Total Kjeldahl Nitrogen
Sulfuric acid digestion of a sample, followed by
measurement of ammonium salt formed using alkaline phenol
procedure or Orion ammonia electrode procedure (Model 95-10)
(Orion, 1971).
Soluble Kjeldahl Nitrogen
Sulfuric acid digestion of a filtered sample, followed
by measurement of ammonium salt formed using alkaline
phenol procedure or Orion ammonia electrode procedure.
Nitrate Nitrogen
Direct analysis of a filtered sample, using the modi-
fied Brucine Method (Jenkins and Medsker, 1964) either
manually or on the Technicon Auto-Analyzer.
Nitrite-N was not determined, as the relative contri-
bution of this species to the total initial N of the
river waters was assumed to be negligible. Hence, the con-
centration of Total N in the samples was computed from the
sum of the initial nitrate-N plus the initial TKN. Detailed
procedures have been described by Sirisinha (1973).
In tests of nutrient regeneration from river water
particles, the river water was filtered through a membrane
filter, and the particles retained on the filters were
scraped into Lake Ontario water, using a metal spatula.
The initial particulate organic-N concentration due to
15
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the river water particles was found by TKN analysis of the
lake water-particle suspension, minus the TKN found in
lake water controls with no river water particles.
NUTRIENT REGENERATION INCUBATIONS
Triplicate 400 ml volumes of whole river water
samples or suspensions of river water particles in lake
water were placed in glass one-liter bottles stoppered
with foam or cotton plugs. Triplicate bottles of lake
water served as controls for the incubations of river
water particles in lake water. All test bottles were
incubated at 20 1 1°C under black plastic sheets in a
walk-in algal culture incubator room. The bottles were
shaken daily and whenever sampled. After various periods
of incubation, aliquots were removed from the bottles
for analyses of ammonia-N and nitrate-N. In some cases,
the ammonia determinations were omitted if the preceding
ammonia concentration was negligible. In the experiments
with suspensions of particles, only the nitrate-N concentra-
tions were determined in the sample particle suspensions
and in the controls. The total incubation period used
in the nutrient regeneration experiments ranged from 35 to
100 days.
A BIOASSAY TEST FOR NITROGEN AVAILABILITY FROM PARTICLES
This algal assay is based on Liebig's law of the
minimum which states that "growth is limited by the
substance that is present in minimal quantity in respect
to the needs of the organism." Increase in absorbance
as a measure of the growth of algae was not used in the
present experiment due to the interference of particles
scattering light. Instead, the growth of algae was
measured by direct algal counting method. Selenastrum
capricornutum was chosen as test organism because it has
16
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several superior qualities as a laboratory organism,
it is solitary and easy to identify, it grows easily in
culture with little variation in form when nutrient con-
ditions are varied, it seems to have low temperature
requirements, it can tolerate both strongly acidic and
alkaline waters (Forsberg, 1972), and it has a rapid
growth (National Eutrophication Research Program, 1971).
According to Murray (1971), light intensity of 500 ft-c
reduced the growth of S_. capricornutum. There was also
evidence of a slight growth rate limitation of Selenastrum
at 200 ft-c in the present experiment. Continuous "cool-
white" fluorescent lighting 400 ft-c - 10 percent was used.
Intensity was measured adjacent to the flask at the liquid
level.
Reagents
Stock and standard nitration solutions: the same as
described in nitrate determination except the range of
standards were from 13.5 gN/ml to 81 ygN/ml.
Synthetic Algal Nutrient Medium (National Eutrophi-
cation Research Program, 1971).
Final concentration of nutrients.
Macro-nutrients - The following salts, reagent grade,
in milligrams per liter of ammonia-free water.
Compound Concentration (mg/1) Element Concentration (mg/1)
K2HP04 1.044 P 0.186
MgCl2 5.700 Mg 2.904
MgS04.7H20 14.700 S 1.911
CaCl2.2H20 4.410 Ca 2.143
NaHC03 15.000 C 1.202
NaN03 25.500 N 4.200
Na 11.001
K 0.469
17
-------
Micronutrients - The following salts, reagent grade,
in micrograms per liter of ammonia-free water.
(Ug/l) (yg/1)
H3B03 185.520 B 32.460
MnCl2 264.264 Mn 115.374
ZnCl2 32.709 Zn 15.691
CoCl2 0.780 Co 0.354
CuCl2 0.009 Cu 0.004
NaMo04.2H20 7.260 Mo 2.878
FeCl3 96.000 Fe 33.051
Na2EDTA.2H20 300.000
Stock solutions.
Macronutrients - Stock solutions of individual salts
might be made up in 1000 times the final concentration.
Micronutrients - The trace metals, FeCl- and EDTA
were combined in a single stock mix at 1000 times final
concentration.
Preparation of medium.
Combination of stock solutions - 1ml of each of the
stock solutions (macronutrients and micronutrients were
added to ammonia-free water to give a final volume of
1 liter. The trace metal -FeClgEDTA mixture (Micronutrients)
was added after filtration.
All media should be stored in dark to avoid any
photochemical changes.
Cell suspension - A 50 ml of fresh culture of S_.
capriconutum in AAP medium with 3x N and P concentration
was used. The tube was covered with parafilm and centri-
fuged at 1200 rpm for 20 minutes. The packed cells were
then washed twice with 10 ml of fresh AAP (-N) medium in
order to get rid of excess nitrogen materials in the
18
-------
medium. These cells were then suspended in 30 ml of new
AAP (-N) medium and counted with a haemocytometer. The
following formula was used to make enough inoculum for
100 flasks, with 104 cells/flask.
U
2700 x 10 cells ml of washed cul-
cell count(cells/ml) of washed culture ~ ture diluted to
100 ml with AAP-N
medium
The concentration of cells after dilution of washed
u
culture to 100 ml should be 27 x 10 cells/ml or 27 x
14
the desired initial population level of 1 x 10 cells/ml.
ij.
Use 1 ml of 27 x 10 cells/ml suspension per bioassay
flask.
Bioassay Test
A 200 ml sample from urban runoff or Lake Ontario
tributaries were filtered through 0.45 micron Millipore
filter, and particles were scraped off into 200 ml of AAP
(-N) medium. A 10 or 20 ml of suspension was removed for
total Kjeldahl-N determination. Volume corrections were
made to get the results of organic-N concentration in
each bioassay flask.
Three sets of five replicates of the following were
performed and incubated at 24 1 1°C, 400 ft-c light
intensity.
Set 1 Blank flasks contained 25 ml AAP (-N) medium
1 ml ammonia-free water
1 ml cell suspension
Set 2 Standard flasks contained 25 ml AAP (-N) medium
1 ml N0~ -N spike
1 ml cell suspension
19
-------
Set 3 Sample flasks contained 25 ml particulate sus-
pension in AAP (-N)
medium
1 ml ammonia-free water
1 ml cell suspension
The shape of the growth curve was found by plotting
the absorbance at 750 nm for standard culture vs time. The
growth plateaus of all standards were obtained from the above
plots. The cells in standards and samples were counted on
the day the plateau was reached. A haemocytometer was used
for cell counting. The concentration of nitrogen vs average
cell counts for the standard was plotted, and the available-N
content of the samples was then calculated.
The initial experiments using the above bioassay method
failed to give satisfactory results, even though the experi-
ments were repeated many times. This failure was due to
contamination by Anabaena, which is the nitrogen-fixing
organism, from the studied particles. Hence, a more refined
bioassay technique was used which gave satisfactory results.
The procedure is described below.
The samples of both urban runoff and Lake Ontario
tributaries after 100 days of incubation were filtered
through a 0.45 micron Millipore filter. Then 20 ml of the
filtrate was used as the nitrogen source instead of the particles.
The experiments were set up as follows:
Set 1 Blank flasks contained 21 ml of deionized water
5 ml of 5 x AAP (-N)
medium
1 ml of cell suspension
20
-------
Set 2 Standard flasks contained 20 ml of deionized water
5 ml of 5 x AAP (-N)
medium
1 ml of nitrate stand-
ards
1 ml of cell suspension
Set 3 Sample flasks contained 20 ml of filtrate
5 ml of 5 x AAP (-N)
medium
1 ml of deionized water
1 ml of cell suspension
Filtration made the sample sterile; neither
Anabaena nor bacteria were present in the filtrate. After
ten days of incubation, the samples were measured at 750 nm.
The growth of algae are shown by comparing with the standard
curves.
21
-------
SECTION VI
RESULTS
INITIAL NITROGEN AVAILABILITY
The analyses of New York rivers (Table A.I, Appendix A)
for initial nitrogen forms showed the following ranges for
"algal-available" N (ammonia-N + nitrate-N) and organic-N,
expressed as a percent of the total N in the samples.
Table 2. INITIAL N AVAILABILITY
Percent of
River Available N
Niagara
Genesee
Oswego
Black
30-67
51-55
44-69
27-58
Total N as:
Organic-N
33-70
45-49
31-56
42-73
No. of Samples
3
4
8
4
Thus, in some of the samples, the extent of nitrogen
mineralization was relatively low, 30 percent or less,
while in other samples, over 60 percent of the total N was
already in an algal-available form by the time of the initial
analyses.
The fraction of total N not present as ammonia-N or
nitrate-N was assumed to be organic-N. Table 3 shows the
initial organic-N concentrations in the samples, as well
as a breakdown of the organic-N into soluble and particulate
fractions, computed by the equations:
22
-------
Soluble organic-N = SKN - Ammonia-N
Particulate organic-N = TKN - SKN
The concentrations of organic N in samples of a given river
were quite variable, as were the relative amounts of
soluble or particulate organic-N. In the Oswego R. samples,
for example, the particulate fraction of organic-N accounted
for 13 to 78 percent of the organic-N.
INCUBATIONS OF RIVER WATERS
The data collected from the individual bottles of
river water during dark incubations are given in Table A.I
of Appendix A. The mean values for ammonia-N and
nitrate-N from each triplicate set of bottles are listed
in Table A.2, Appendix A. The sum of these mean values
estimated the algal-available N as a function of time
during the incubations; these sums are given in Table A.2,
expressed in concentration units and as percents of the
total N in the sample.
Ammonia-N levels generally dropped to the analytically
detectable limit of 0.05 mgN/1 in 25 to 50 days. Values
of ammonia-N of less than 0.05 mgN/1 were considered as
equal to zero in all calculations of percentage avail-
ability. Formation of nitrate from the ammonia appeared
to be completed by 25 to 50 days in most samples.
Table 4 presents the results of the dark incubations
in terms of the maximum algal available N values
observed after 35 to 50 days of incubation.
The Niagara R. samples showed 54 to 91 percent of
their total N available as ammonia plus nitrate after the
35 or 50 days of dark incubation. Figure 3 illustrates
the changes in available N as a function of time. Samples
No. 41 and50 showed very similar behavior, while Sample Kb..56
23
-------
Table 3. DISTRIBUTION OF ORGANIC NITROGEN FORMS IN
RIVER WATER SAMPLES
Soluble Organic-N Particulate Organic-N
Organic-N
Sample No. (mgN/1) (mgN/1) (%Organic-N) (mgN/1) (%Organic-N)
Niagara R.
50
56
Genesee R.
34
42
51
58
0.15
0.31
0.79
1.08
0.68
0.59
1.02
0.06
0.18
0.59
0.37
0.17
0.15
0.44
40
58
75
34
25
25
43
0.09
0.13
0.20
0.71
0.51
0.44
0.58
60
42
25
66
75
75
57
Oswego R.
28
31
35
43
47
48
52
59
Black R.
36
44
53
60
0.55
0.39
0.39
0.68
0.45
0.47
0.83
1.04
0.42
0.25
0.48
0.91
0.45
0.34
0.33
0.15
0.30
0.20
0.24
0.59
0.29
0.10
0.30
0.56
82
87
85
22
67
43
29
57
69
40
62
62
0.10
0.05
0.06
0.53
0.15
0.27
0.59
0.45
0.13
0.15
0.18
0.35
18
13
15
78
33
57
71
43
31
60
38
38
appeared to be significantly lower than the other two samples
with regard to its nitrogen mineralization, and relatively
higher with regard to total N concentration (Table 4).
The Genesee R. samples had maximum available N values of 60
to 75 percent of total N. The samples with the lowest values
were Nos. 34 and 58, both of which had relatively high total N
values compared to Samples No. 42 and 51 (Table 4). Figure
24
-------
lOOr
SAMPLE NO.
20 40
TIME, DAYS
60
Figure
3. Percent of total Nas ammonia-N +
nitrate-N during dark incubation of
Niagara R samples
25
-------
4 shows the close agreement between Samples No. 42 and
51 and between Nos. 34 and 58.
Oswego R. waters (Figures 5 and 6 ) showed maximum
availability values of 58 to 91 percent of total N, although
most of the samples had values between 58 and 82 percent.
The lowest availability was seen in Sample No. 59 which also
had the highest total N concentration (Table 4 ). The
rather sharp increases in available N seen after 64 days in
Samples No. 31 and 35 (Figure 5 ) were probably analytical
errors. The curves had essentially reached a plateau by day
64, after which no further changes would be expected.
The Black R. samples (Figure 7 ) demonstrated consistent
maximum availability values of 67 to 75 percent of total N
in three samples, while in the third sample (No. 60), the
availability was low, only 36 percent at its maximum value.
This sample also had the highest total N content of the sample
set (Table 4).
In an attempt to determine whether a constant fraction
of the organic forms from a given river were being mineralized
in the dark incubations, the maximum values of ammonia-N
plus nitrate-N given in Table 4 were expressed as a percent
of the organic-N in the sample and tabulated in Table 5.
The most outstanding feature of the data in this table is the
relationship of percent availability to the initial concentra-
tion of organic-N. Those samples high in total N con-
centrations and low relative total N availability appeared to
have high initial organic-N concentrations and low organic-M
availability. With the exception of Sample No. 34, all of
the samples with high organic-N concentrations were collected
on the June 16-17 sampling trip.
As a check on the nitrogen mass balance of the samples
during the incubations, the initial total N values of some
samples were compared to the total N value in those samples
26
-------
Table 4. MINERALIZATION OF TOTAL N TO AMMONIA
AND NITRATE IN 35-50 DAYS
Sample No.
Niagara R.
41
50
56
Genesee R.
34
42
51
58
Oswego R.
28
31
35
43
47
48
52
59
Black R.
36
44
53
60
Initial
mgN/1 of
total N
0.45
0.82
1.13
2.21
1.52
1.26
2.26
1.34
1.27
1.16
1.42
1.15
1.01
1.49
2.30
0.89
0.59
0.75
1.25
Maximum
NH3-N +
mgN/1
0.41
0.74
0.61
1.33
1.11
0.95
1.35
1.04
1.00
1.06
0.98
0.87
0.83
1.04
1.32
0.62
0.44
0.50
0.45
observed
NO" -N
% total N
91
90
54
60
73
75
60
78
79
91
69
76
82
70
58
70
75~"
67
36
Total time
of incubation,
days
35
50
50
50
35
50
50
50
50
50
35
50
50
50
50
50
35
50
50
27
-------
Table 5. MINERALIZATION OF ORGANIC-N TO
AMMONIA AND NITRATE IN 35-50 DAYS
Sample No.
Niagara R.
41
50
56
Genesee R.
34
42
51
58
Oswego R.
28
31
35
43
47
48
52
59
Black R.
36
44
53
60
Initial mgN/1 as:
NH3-N+
Organic-N NOg-N
0.15
0.31
0.79
1.08
0.68
0.59
1.02
0.55
0.39
0.39
0.68
0.45
0.47
0.83
1.04
0.42
0.25
0.48
0.91
0.30
0.51
0.34
1.13
0.84
0.67
1.24
0.79
0.88
0.77
0.74
0.70
0.54
0.66
1.26
0.47
0.34
0.27
0.34
Maximum Increase in
NH3-N + Available N
N03-N
0.41
0.74
0.61
1.33
1.11
0.95
1.35
1.04
1.00
1.06
0.98
0.87
0.83
1.04
1.34
0.62
0.44
0.50
0.45
mgN/1
0.11
0.23
0.27
0.20
0.27
0.28
0.11
0.25
0.12
0.29
0.24
0.17
0.29
0.38
0.08
0.15
0.10
0.23
0.11
% of Organic-N
73
74
34
18
40
48
11
46
31
74
35
38
62
46
8
36
40
48
12
28
-------
lOOr
20
40 60
TIME .DAYS
80
100
Figure 4.
Percent of total N as ammonia-N + nitrate-N during
dark incubation of Genesee R. samples
29
-------
100 r
SAMPLE NO.
Figure 5.
20 40
TIME, DAYS
Percent of total Nas ammonia-N +
nitrate-N during dark incubation
of Oswego R. samples
30
-------
SAMPLE NO.
lOOr
20
40 60
TIME,DAYS
80
100
Figure
6. Percent of total N as ammonia-N + nitrate-N during
dark incubation of Oswego R. samples
31
-------
100 r
SAMPLE NO.
40 60
TIME, DAYS
80
100
Figure 7. Percent of total Nas ammonia-N + nitrate-N during
dark incubation of Black R. samples
3?
-------
after incubation. In both cases, the total N was computed
from the sum of TKN plus nitrate-N, neglecting Nitrite-N.
Table 6 shows the results of the comparisons. Ex-
cept for Samples No. 31 and 35, the final and initial
values appeared to agree as closely as could be expected
from analytical considerations. The final values from
Samples No. 31 and 35 appeared to be higher than the
initial values by a significant amount.
Table 6. NITROGEN MASS BALANCES
Sample
28
31
35
43
42
41
44
Initial total N
(mgN/1)
1.34
1.27
1.16
1.42
1.52
0.45
0.59
Final total N
(mgN/1)
1.28
1.49
1.59
1.33
1.44
0.52
0.65
Incubation
(days)
82
64
64
35
35
35
35
MINERALIZATION OF PARTICULATE ORGANIC N
Tables A.3 through A.4 of Appendix A give the detailed
data from the individual incubations of river water particles
in lake water. Figure 8 shows the average production of
nitrate-N from particles of Genesee R. (No. 42) water incu-
bated in lake water from Station No. 10 . Only 26
percent of the organic-N in the particles was re-
leased to solution as nitrate-N. The mineralization appeared
to be complete after about 14 days. A similar test with
Oswego R. (No. 43) particles in the same lake water showed
about 75 percent conversion of organic-N to nitrate-N after
about 21 days (Figure 9).
33
-------
0.5r
0.4
0.3
i
UJ
0.2
O.I
O— O NET MEAN NITRATE-N
FROM PARTICLES{mgN/l)
NET MEAN NITRATE-N
FROM PARTICLES ( % OF
PARTICULATE ORGANIC N)
10 20
TIME,DAYS
30
-i 100%
o
z
80
60 I
u.
o
40 £
UJ
o
a:
UJ
a.
20 '
UJ
£T
H
Z
Figure 8. Nitrate-N production from Genesee R. (42 )
particles in Lake Ontario (10) water
-------
0.5
0.4
o»
£0.3
Z
I
UJ
h-
0.2
O—O NET MEAN NITRATE-N
FROM PARTICLES (mgN/1)
NET MEAN NITRATE-N
FROM PARTICLES ( % OF
PARTICULATE ORGANIC N)
0 I
100%
z
o
z
o
80 o:
o
UJ
i-
40
20
u_
o
z
UJ
o
(T
UJ
Q.
UJ
<
tr
10 20
TIME, DAYS
30
Figure 9.
Nitrate-N production from Oswego R.(43)
particles in Lake Ontario (10) water
35
-------
In lake water from Station No. 96 samples of
Genesee River (No. 58) and Oswego River (No. 59) par- •
tidies released 67 and 108 percent, respectively, of their
organic-N as nitrate-N (Figures 10 and 11) after 25 days.
MINERALIZATION OF URBAN RUNOFF SAMPLES
The chemical characteristics for nitrogen species
analyses are given in Table 7. Thirteen urban runoff
samples collected from Madison, Wisconsin areas were per-
formed for the long-term study of mineralization. A period
of 82-100 days was selected for this study. All of the sam-
ples for mineralization study were run in triplicate and
were performed at 21- 1 C under aerobic conditions in the
dark. The range obtained in the analysis of incubated trip-
licate bottles were shown in the figures. Assuming no occur-
rence of photosynthesis and bacterial nitrogen fixation, the
total Kjeldahl nitrogen value from day zero was used to cal-
culate total-N. Percent availability was calculated from
NHL -N and N07 -N each day that a sample was taken. Figures
12 to 15 demonstrated the mineralization of total
Kjeldahl-N to NH^ -N and N0~ -N of runoff water from the
Whitney Way station at different days of rainfall. Although
different concentrations of nitrogen forms were found at
day zero, all the curves showed the same trend of nitrogen
forms changing. In Figures 12, 14, and 15 there was an
immediate increase in the NH^ -N concentration at day 10 of
incubation while in Figure 13 this happened at day 25. After
that, the NHU -N in solution decreased to 0.05 mgN/1 by the end
of the study. The NO, -N increased steadily in all samples.
However, a sharp increase in concentration was observed at dciy
25 in Figures 12 and 14, at day 50 in Figure 13 and at day
82 in Figure 15. The concentration of both soluble and
36
-------
Table 7. INITIAL CONCENTRATIONS OF SELECTED NITROGEN
SPECIES IN THE SAMPLES USED IN THIS
INVESTIGATION
Samples
Madison
A- 6
A- 8
A- 9
A-12
B-6
B-8
B-9
B-12
D-6
D-8
D-10
D-12
F-9
NH* -N
Date Collected (mgN/1)
Urban Runoff
10/20/72
12/30/72
1/17/73
3/5/73
10/20/72
12/30/72
1/17/73
3/5/73
10/20/72
12/30/72
1/18/73
3/5/73
1/17/73
Samples
0.13
0.59
0.80
1.12
0.14
0.75
0.82
1.32
0.08
0.64
0.51
1.07
0.85
N0~ -N
(mgN/1)
0.02
0.45
0.21
0.71
0.005
0.62
0.31
0.30
0.24
0.80
1.56
0.335
0.63
Total
Kjeldahl-N
(mgN/1) pH
1.85
1.60
2.40
4.28
3.60
1.50
2.30
4.24
2.45
1.70
2.70
4.60
1.60
37
-------
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o
N
o
00
o
04
JD
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in
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o.
00
in
o>
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to
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ro OJ
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JO lN30d3d'N-31V«llN
>-
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a.
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a:
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39
-------
2.0
O
Z
UJ
o
O—O NH^-N
A—A NOj-N
D—•D PERCENT AVAILABILITY
0
Figure 12. The mineralization of runoff water from Whitney Way
station collected on October 20,1972 incubated under
aerobic conditions
2.0
z
o
oc
H
z
u
8z i.o
o>
o
cr
O—O NH^-N
A—A NOj-N
D—D PERCENT AVAILABILITY
100
80
60
40
m
<
0 10
25
50
DAYS
100
Figure 13. The mineralization of runoff water from Whitney Way
station collected on December 30, 1972 incubated
under aerobic conditions
-------
4.0
<
cc
I-
z
UJ
o —
z ^
o z
O 01
zl
UJ
o
o
3.0
1.0
O O NH+-N
A—A NOg-N
D—D PERCENT AVAILABILITY
100
80
S=!
U ffi
„ o: <
60 uj _j
a. 5
40 ^
10
25
50
DAYS
100
Figure 14. The mineralization of runoff water from Whitney Way
station collected on January 17, 1973 incubated under
aerobic conditions
O—O NH+-N
4.0r ^—A
z
o
3.0
z
UJ
o —
z ^
oz 2.0
VJ w*
z£
Ul
2
1.0
0Li
-N
D — D PERCENT AVAILABILITY
Figure
1 80
60
40
20
O GO
10
25
50
DAYS
100
15. The mineralization of runoff water from Whitney Way
station collected on March 5, 1973 incubated under
aerobic conditions
-------
total Kjeldahl-N decreased at the end of the experiment.
The percent availability of all figures were found to
increase steadily throughout the incubation period.
Figures 16 to- 19 showed the mineralization of total
Kjeldahl-N to NH^ -N and N0~ -N of runoff water from the
Manitau Way station on different days of rainfall. The
soluble and total Kjeldahl-N decreased at the end of the
-f
study. A sharp increase in NH^ -N was observed in Figures
16 and 19 at day 10. It then decreased to 1.0 mgN/1 at
day 10 and declined to 0,10 mgN/1 at the end of the study.
The NH^ -N in Figures 17 and 18 gradually declined over
the period of 100 days. There was a slow increase in N0~ -N
of all samples throughout the experiment. The percent
availability increased until the end of the experiment.
The results of the study of runoff water from the
Water Chemistry Station are shown in Figures 20 and 23.
There was an increase in NH^ -N at day 10 in Figure 20 and
23; after that there was a decrease throughout the experi-
ment. No change occurred in N0~ -N in Figure 20 at day 10
while in Figure 23, a sharp increase was observed. However,
after day 10 both figures showed the increase in NOQ -N until
+ •=
the end. There was a decrease in NEL -N in Figure 21 and
22 from the beginning to the end of the experiment. An
increase in NO^ -N in Figure 22 was observed from day 0
to day 100 while in Figure 21 a decrease was found at
day 25. The soluble and total Kjeldahl-N also decreased at
the end of the study. The percent availability increased
throughout the incubation time.
The results obtained from the Stone Ridge Apartment
station were demonstrated in Figure 24. The NH^ -N in-
creased at day 10 and after that decreased. The NO, -N had
-------
4.0p o—O NH+-N
3.0
Z
LU
O C
Z v.
LU
O
O
(T
1.0
A—A NOj -N
D—D PERCENT AVAILABILITY
100
80
60
ir <
LJ _l
20°-<
Figure 16. The mineralization of runoff water from Manitau Way
station collected on October 20,1972 incubated under
aerobic conditions
z
O
<
cr
2.0
1.5
LU
O C
O
1.0
LU
O
O
a:
0.5
O O NH^-N
A—A NOs-N
D—D PERCENT AVAILABILITY
10
25
50
DAYS
100 Kt
Z -J
80 o
60
10
Figure 17. The mineralization of runoff water from Manitau Way
station collected on December 30,1972 incubated
under aerobic conditions
-------
4.0
ft 3.0
UJ
o ---
§!2-°
Z E
ui —
ft
h-
1.0
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A—A NOj -N
D—D PERCENT AVAILABILITY
10
25
50
DAYS
100
100
80 h£
60 o ffi
UJ-J
40 a-5
Figure 18. The mineralization of runoff water from Manitqu Way
station collected on January 17,1973 incubated under
aerobic conditions
4.0
cc
z
UJ _
o —
z "
3.0
LU
o
O
cr
2 O
<:-u
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A—A NOj -N
D—D PERCENT AVAILABILITY
OL i
10
25
50
DAYS
100
90
80 i
70
60
50
82
Figure 19. The mineralization of runoff water from Manitqu Way
station collected on March 5,1973 incubated under
aerobic conditions
-------
4.0
3.0
z
UJ
oc
2 >»
Oz
Ul
o
o
ac.
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A—A N03~-N
D—D PERCENT AVAILABILITY
80
60 Ht
Z_j
40 o"
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iud
20 °-^
<
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10
100
Figure 20. The mineralization of runoff water from WaterChemistry
lab station collected on October 20,1972 incubated
under aerobic conditions
o—o NH£-N
A—A NO, -N
2.0r- D—n PERCENT AVAILABILITY
<
a:
ui ^.
•z. ~
SSi-o
= t
UI
o
o
(T
0L
80
60 UOD
(T <
UJ-I
40 o-
10
25
50
DAYS
100
Figure 21. The mineralization of runoff water from WaterChemistry
lab station collected on December 30,1972 incubated
under aerobic conditions
-------
z
o
(T
t-
z
LJ
LU
O
O
(T
4.0
3.0
2.0
1.0
O—O NH4-N
A—A N03~-N
D—D PERCENT AVAILABILITY
QL I
10
25
50
DAYS
60
UJ-J
40
100
Figure 22. The mineralization of runoff water from Water Chemistry
lab station collected on March 5, 1973 incubated under
aerobic conditions
<
cr
4.0
3.0
LJ
O C
I I2'0
LU
(5
O
ft
t-
1.0
O—O NH^-N
A—A N0~ -N
D—D PERCENT AVAILABILITY
10
25
50
DAYS
80
6O
40
20
O CD
100
Figure 23. The mineralization of runoff water from Water Chemistry
lab station collected on March 5, 1973 incubated under
aerobic conditions.
-------
z
o
a:
K
Z
UJ
3.0
2.0
oz
o o»
E
UJ
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O ONH^-N
O—O N03" -N
-------
a continuous increase until the end. The soluble and total
Kjeldahl-N decreased as the percent availability increased.
The results of the triplicate run of each sample were
very similar in their mineralization pattern.
BIOASSAY TEST FOR AVAILABILITY TO ALGAL GROWTH FROM PARTICLES
The algal bioassay test was set up to directly deter-
mine the fraction of organic-N available for algal growth.
The particles from both urban runoff and Lake Ontario tri-
butaries were suspended in AAP (-N) medium and incubated at
24 ± 1 C, 400 ft-c light intensity. Selenastrum capricornutum
was used as the test organism. Standards and blanks were
also run. Figure 25 shows the growth curve of Selenastrum
using various nitrate concentrations as the source of nitro-
gen. The growth of algae was measured at 750 nm and was
plotted versus day of incubation. The cells in both the
samples and the standards were counted on day 21. There was:
not much growth of cells in sample flasks; however, the nitro-
gen fixing blue-green algae, Anabaena, were found in large
amounts compared to Selenastrum. The experiments were re-
peated. The incubation time was changed to 7 days hoping
that the Selenastrum would grow faster than the Anabaena.
On day 7, the samples were counted, no Anabaena were present
but there was also no growth of Selenastrum. The samples
were counted every three days. Finally Anabaena was found
to be present in the samples on day 14 and no growth of
Selenastrum was observed. Another experiment was then
performed to solve the problem of Anabaena contamination
from particles. The filtrate from 100 days incubation samples
was used as a nitrogen source instead of the particles. The;
results of this study are presented in Table 8. The in-
crease of absorbance which is related to the growth of
Selenastrum was observed in all samples.
48
-------
Table 8. THE NITROGEN AVAILABILITY FROM PARTICLES
USING BIOASSAY TEST AT 10 DAYS
Absorbance Concentration
Samples at 750 nm mgN/1
A-12-1 0.25 2.15
2 0.25 2.15
3 0.26 2.20
B-12-1 0.33 >4.00
2 0.32 >4.00
3 0.33 >4.00
D-12-1 0.32 >4.00
2 0.33 >4.00
3 0.32 >4.00
28-1 0.10 0.75
2 0.11 0.85
3 0.11 0.85
41-1 0.045 0.30
2 0.045 0.30
3 0.045 0.30
42-1 0.11 0.85
2 0.11 0.85
3 0.11 0.85
43-1 0.10 0.75
2 0.095 0.73
3 0.10 0.75
44-1 0.05 0.35
2 0.05 0.35
3 0.05 0.35
49
-------
4000/xg/l
12 15 18 21
Blank
24
Figure 25. The growth curve of Selenastrum capricornutum using
nitrate as nitrogen source
50
-------
SECTION VII
DISCUSSION
LAKE ONTARIO SAMPLES
River water samples containing organic N forms from
runoff, domestic sewage, industrial wastes, and aquatic
organisms were incubated in darkness to promote nitrogen
mineralization by the bacteria in the samples. Under
such conditions, bacterial populations have been shown
to rise in two to three days to levels 103 to 10 times
the populations in natural waters (Renn, 1937). Conse-
quently, the rate of inorganic N production from organic
N sources in such tests was related to the rate of bacte-
rial growth, the length of the viable period, and the
rate of autolysis after death. (Renn, 1937). This rate
may not be the same as the rate in natural waters because
of the different bacterial population densities. However,
the objective of the tests was to estimate the long-term
susceptibility of the organic N in the rivers to mineral-
ization, with the assumption that any organic N which
was mineralized in the dark incubation could eventually
produce available N for algal growth in Lake Ontario.
The results of these tests are intended to be the upper
bounds on the nitrogen mineralization in each sample.
Although the nitrite-N was assumed negligible in
computation of the final reported results, the error from
this assumption was probably small. Nitrate values
51
-------
generally showed little change after 25 to 50 days, indi-
cating that all nitrite had been converted to nitrate.
Since only the maximum value of the sum of nitrate plus
ammonia was selected for the final reported result of an
incubation, the error from neglecting nitrite-N was mini-
mized. The reported values of (ammonia-N + nitrate-N) cit
incubation times of less than 25 days (Figures 3-7)
may have been lower than the true "available -N", however,
because of nitrite production from ammonia during the
initial stages of the incubations.
The apparent increases in total N during the incu-
bations of Samples 31 and 35 for 64 days (Table 6) were
probably the result of neglecting nitrite-N-in the calcu-
lation of initial total N. Since any nitrite initially
present would be converted to nitrate during the incu-
bation, the final total N value (NOs-N + TKN) would in-
clude the contribution from nitrite and be larger than
the initial total N value. The values given in Table 4
should probably be based on the final total N values, in
the case of Samples 31 and 35. Recalculation of the
available N percentages yields the value of 67 percent
for both samples instead of the values of 79 and 91
percent shown in Table 4.
Other workers have noted an increase in total N
during regeneration studies of nitrogenous organic N (Von
Brand et al. , 1937, Sawyer et_ al. , 1944). Such an in-
crease could be the result of nitrogen fixation by bac-
teria. A sample of Oswego R. water (59) which had been
incubated for 33 days was tested for N-fixation by an
acetylene reduction procedure and found to have a fix-
ation rate of 2.6 ygN/1 per week (Lonergan, 1973), or
0.024 mgN/1 for a 64 day incubation period. This amount
of nitrogen is much less than the increases shown by
52
-------
Samples 31 and 35 in Table 6 (0.22 to 0.43 mgN/1). A
similar test on Sample 58 from the Genesee R. indicated
N-fixation of 0.039 mgN/1, predicted for a 64-day incu-
bation. Black R. Sample 60 showed no N-fixation.
The incubations of unfiltered river water demon-
strated that a large percentage of the total N in the
rivers should be considered eventually available for
algal growth in Lake Ontario. In many samples the high
availability was related to the high concentrations of
ammonia and nitrate initially present. These "readily
available" N forms were the result of (1) direct inputs
from runoff and wastewaters, (2) mineralization of
organic N between the point of discharge to the river
and the sampling station near Lake Ontario, or (3) min-
eralization in the samples prior to their initial analysis
in Madison. With the exception of Sample 60 (Black R.),
all samples showed over 50 percent of their total N to
be subject to mineralization or already mineralized. The
wide range of values and the small number of samples test-
ed from each river preclude any one estimate of nitrogen
availability for a given river. Rather, the data (Table
4), should be considered as temporary upper and lower
bounds on N availability until more detailed data are
available. Thus, the data from Table 4 can be summa-
rized as follows:
River Range of Availability (% of Total N)
Niagara 54-91
Genesee 60-75
Oswego 58-91
Black 36-75
Use of these percentages in conjunction with a nutrient
budget based on total N loadings from each river would
provide estimates of the available N loadings from the
rivers.
53
-------
Another approach to the correction of total N load-
ing data to an available N basis would use the following
equation:
Available N = Anunonia-N + Nitrite-N + Nitrate-N +(Org.-N)
X (% Org.-N Avail.)
where the nitrogen forms are measured in samples collect-
ed near the mouth of the river. The data in Table 5
indicated, however, that the percent of organic-N which
could be considered available (% Org.-N Avail.) may be
extremely variable and related to the concentration of
organic matter in the sample. Those samples with the
highest organic-N in each set of samples appeared to have
the lowest percent of organic N available. As boundary
conditions on organic-N availability, the data from
Table 5 can be summarized as follows:
River Range of Availability (% of Org. -N)
Niagara 34-74
Genesee 11-48
Oswego 8-74
Black 12-48
It is important to note that these data were collected
during the spring flow period only, and not during other
seasons of the year, when the quantity and quality of
the organic-N inputs may be different, particularly with
respect to agricultural runoff sources. Consequently,
these estimates are only valid for the spring flow period
and the N loadings carried by the rivers to Lake Ontario
during this period.
As much as 78 percent of the organic-N in some of
the samples was present as particulate organic-N. This
operationally defined class may have also included some
ammonia bound to the surface of soil particles. The
eventual fate of particulate N forms in Lake Ontario is
difficult to predict, since it depends upon physical
54
-------
factors such as particle settling rate and rate of sub-
sequent deposition of covering layers of sediment. The
tests with unfiltered water attempted to reduce these
physical factors to negligible levels by providing fre-
quent swirling of the test bottles. Biological factors
were investigated directly by incubating membrane-filter-
able particles in lake water. The two Genesee R. samples
tested showed different particulate organic-N availabili-
ty than was shown by the total organic-N (soluble plus
particulate) in each sample (from Table 5) :
Percent availability of:
Sample No. Org.-N Part. Org.-N
42 40 26
58 11 67
The same analysis of the Oswego R. samples showed:
Percent availability of:
Sample No. Org.-N Part. Org.-N
43 35 75
59 8 108
If it is assumed that soluble organic compounds are more
readily decomposed than are particulate forms, then the
availability of organic-N should always be greater than
the availability of particulate organic-N, since organic-
N includes both soluble and particulate forms. The data
above show the opposite for all but Sample 42. In the
case of Sample 59, if 100 percent of the particulate
organic N in the sample (0.45 mgN/1) were available, as
indicated above, the available percent of organic-N would
be at least 0.45/1.04 X 100 = 43 percent. The value
determined from incubation of unfiltered water was only
eight percent, however. The same type of disparity was
seen with Samples 58 and 43.
55
-------
Since some of the particulate organic-N in these
samples may have been in the form of planktonic organ-
isms, some comparisons are possible with the data from
other workers who have investigated the decomposition of
aquatic organisms. Waksman et al. (1931) found that
marine zooplankton residues yielded over 50 percent of
their total N as ammonia during bacterial decomposition.
Golterman (1960) performed sterile lysis of Scenedesmus
quadricauda by ultraviolet radiation or chloroform treat-
ment and reported only 20-30 percent release of the N
compounds. The remaining N forms, present as insoluble
nucleic acids and proteins, were suspended in lake water
and allowed to decompose. About half of the N in the
residue was converted to ammonia in five days, for an
overall mineralization percentage of 55-70 percent. These
data are similar to the percentages of organic N availa-
bility found for Samples 58 and 43, of 67 and 75 percent,
respectively. Somewhat lower figures were reported by
Foree and Barrow (1970), who found about 54 percent of
the initial particulate N remaining in mixed cultures of
algae, indigenous bacteria, and zooplankton after 200
days of aerobic decomposition at 20°C. This would rep-
resent a percent availability of 46 percent of the
particulate N.
URBAN RUNOFF SAMPLES
The average concentration of nitrogen species from
the triplicate bottles of urban runoff from different
stations in Madison areas were summarized in Table 9.
The highest increase in NH^-N on day 10 of Samples
A-6, A-9, B-6, B-12, D-6, and D-12 and on day 25 of
Samples A-8, A-12 and B-8 indicated the gradual conversion
of the organic-N to NHiJ-N. After that under aerobic
56
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— —
condition NH^-N will be oxidized to NO^-N and NO~-N, re-
spectively. This agreed with the increase in NO~-N while
+ d
NH^-N decreased after day 10 in the samples. As expected,
NOQ-N accumulated in all the experiments and reached the
. d +
highest value on the final day of incubation while NEL-N
approached zero by the end of the study. At day 100, the
decrease in total Kjeldahl-N in all runoff samples in-
cubated at 21°C under aerobic condition supported the
evidence that organic-N mineralization was occurring in
the studied system.
The same results were also obtained from the experi-
ment on the sediments of Wisconsin lakes (Chen et al.,
1972). Nitrification occurred after a 3-day lag phase
in Chen's experiment instead of 10 days in this experi-
ment. However, in all cases, onset of nitrification was
accompanied by a corresponding decline in soluble and ex-
changeable NEL-N. He also observed an effect of tempera-
ture on nitrification rate. The rate of nitrification
was increasing 2.7 times on a 15°C temperature increase.
Thus, an even higher rate of nitrification would be ex-
pected to occur in shallow water such as the rivers and
the epilimnion of the lakes than in deep water where
higher water temperatures exist.
No change in NHU-N on Samples B-9, D-8, and D-10 was
observed on day 10 or day 25. This may be due to the
equal rate of mineralization and nitrification in these
samples. The increase in NOl-N in these three samples
O
also agreed with this explanation.
The average values of NH^-N, NOl-N and total
Kjeldahl-N at each station were also calculated and are
presented in Table 9.
The average NH^-N of all three stations increased
at day 10 and decreased after that. The NOl-N gradually
58
-------
increased until the end of incubation. The total
Kjeldahl-N of all three stations (A,B, and D) decreased
at the end. The patterns of mineralization at different
stations (Figures 26 through 28 ) were the same and in-
dicated no effect of land usage on mineralization patterns.
No difference of mineralization patterns between the
urban runoff samples and Lake Ontario tributary samples
were noticed in this study. However, the total Kjeldahl-
N in Lake Ontario tributary waters were very low compared
to the urban runoff values. This showed that the initial
total Kjeldahl-N in each sample had no effect on miner-
alization patterns.
The mineralization patterns of particles were ob-
served to be the same as of water from urban runoff and
Lake Ontario tributaries. This indicated that both solu-
ble organic-N and particulate organic-N could be mineral-
ized to NH^-N and followed by nitrification under aerobic
conditions.
The concentration of NOl-N, NEL-N and organic-N in
each sample at day zero and day 100 were summed up to get
the total-N. The increase in total nitrogen at day 100
was observed in most of the samples. Theoretically,
in the closed system study, the total-N content should
be constant. Only transformation of nitrogen occurred.
The increase in total-N in the studied system could be
explained by the fact that some bacteria (green and
purple bacteria, the azotobacter group, and some
anaerobic spore formers) could convert dissolved gaseous
nitrogen to NH3 (Kluesener, 1972). This agreed with
Sawyer et al. (1944) who found a increase of 11.2 mg
59
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total-N in 188 days in an incubated sample of 90 percent Lake
Mendota water mixed with 10 percent sewage effluent. This
amount yielded a production of 0.06 mgN/day. The wide dis-
tribution of N-fixing organisms in Lake Mendota was also found
by Tew (1959).
Von Brand et_ al. (1937) investigated the regeneration of
nitrogenous organic matter in sea water at room temperature
in the dark. From measurements of the disappearance of ni-
trogen in the decomposing plankton material and the appear-
ance of nitrogen in the form of NH^ -N, NOl -N and N03 -N in
the water, he also found a continuous increase in total-N
throughout the experiment.
BIOASSAY TEST FOR AVAILABILITY
The particles from Lake Ontario tributaries and urban
runoff were incubated with Selenastrum in AAP(-N) medium to
study the availability of nitrogen. The negative results
with the growth of Selenastrum in this experiment were ob-
served. However, it cannot be concluded that the tested par-
ticles did not serve as sources of nitrogen because it was
a short-term study. The failure might be due to the fact that
the rate of mineralization was not rapid enough to supply ni-
trogen to the tested algae in a medium with no nitrogen.
Anabaena, N-fixing blue green algae, was observed in the cul-
ture after day 14 of incubation. This blue-green algae was
thought to be originally present in the particles. Under the
conditioned study, every necessary nutrient for algae growth
was present except nitrogen; however, these blue-green algae
could fix nitrogen from the atmosphere resulting in the mas-
sive growth of them in the tested sample while the Selenastrum
could not grow.
The later experiment performed on the filtrate after the
end of mineralization gave the successful results in the growth
61
-------
of tested algae as shown in Table 8. This showed that
4. _
the NH^ -N and N03 -N resulting from the mineralization
and nitrification could be available for algal growth.
The average concentration of nitrogen in the filtrate per
studied flask was shown in Table 10. The growth of algae
which related with the absorbance was correlated well with
the nitrogen concentration in the filtrate. However, the
average availability of nitrogen measure by chemical test
was shown to be less than the result obtained from bioassay
test. It is likely that the algae is able to use soluble
organic-N besides NEL -N and NOl -N in the filtrate for
growth.
Table 10.
COMPARISON OF NITROGEN AVAILABILITY FROM CHEMICAL
AND BIOASSAY METHOD
Samples Chemical N-availability Absorbance Bioassay N-avail-
(mgN/1) ability (mgN/1)
per flask per flask
A-12
B-12
D-12
28
41
42
43
2.85
3.33
2.73
0.81
0.27
0.76
0.69
0.31
0.25
0.33
0.33
0.11
0.045
0.11
0.10
0.05
2.15
> 4.00
> 4.00
0.85
0.30
0.85
0.75
0.35
62
-------
AVAILABILITY STUDIES
The short-term availability was considered to be more
important than the long-term availability in the lakes. In
this stady, short-term availability was defined as the
amount of NH^ -N and N0~ -N that was found to be present
in the interstitial water of samples in zero to 10 days of
incubation. The long-term availability referred to the
incubation time of 50-100 days.
In Table n, the average percent nitrogen availabi-
lity in short-term (10 days) and long-term (100 days) stu-
dies of urban runoff from different stations in the Madison
areas are summarized. The average percent availability
from residential area station A, B, and D was found to be
very close together in the range of 47-55 percent for short-
term study and in the range of 72-97 percent for long-term
study. If the estimated annual amount of nutrients enter-
ing Lake Mendota from urban area (4.6 Ibs/acre/yr) (Son-
zogni and Lee, 1972) was applied, the amount of nitrogen
to be available for plants after entering the lake in 10
days would be in the range of 2.2-2.5 Ibs/acre/yr.
63
-------
TABLE 11. SUMMARIZATION OF THE PERCENT N-AVAILABILITY IN SHORT-TERM
AND LONG-TERM STUDIES OF URBAN RUNOFF
Samples Zoning Code Short-term
A-6-1 Rl
2
3
A-8-1 Rl
2
3
A- 9-1 Rl
2
3
Overall Average of
B-6-1 R2
2
3
B-8-1 R2
2
3
B-9-1 R2
2
3
Overall Average of
D-6-1 R5
2
3
D-8-1 R5
2
3
D-10-1 R5
2
3
Percent
Availability
40
42
40
54
55
54
67
76
69
Station A
30
31
33
67
66
67
70
60
44
Station B
37
31
29
61
54
58
51
55
50
Average
41
54
71
55
31
66
58
52
32
58
52
Long-term
Percent
Availability
95
92
86
90
89
97
110
110
107
80
82
61
95
93
95
105
105
103
83
80
51
76
74
74
72
73
Average
91
92
109
97
74
94
104
91
71
75
72
Overall Average of Station D 47 70 72
-------
REFERENCES
Austin, E.R. 1970. The Release of Nitrogenous Compounds
from Lake Sediments. M.S. Thesis. University of
Wisconsin, Madison. 61 p.
Chen, R.L., D.R. Keeney, and J.G. Konrad. 1972. Nitri-
fication in Lake Sediments of Selected Wisconsin
Lakes. J. Environ. Quality. 1. (2): 151-154.
Foree, E.G. and R.L. Barrow. 1970. Algal Growth and
Decomposition: Effects on Water Quality, Phase II.
Nutrient Regeneration, Composition, and Decomposition
of Algae in Batch Culture. Res. Rep. No. 31. Univ.
of Kentucky Water Resources Inst., 90 p.
Forsberg, C. G. 1972. Algal Assay Procedure. Jour. WPCF.
44_ (8): 1623-1628.
Gerloff, G.C. and F. Skoog. 1957. Nitrogen As a Limiting
Factor for Productivity for the Growth of Microceptis
aeruginosa in Southern Wisconsin Lakes. Ecology. 38:
556-561.
Golterman, H.L. 1960. Studies on the Cycle of Elements in
Fresh Water. Acta Botanica Neerlandica. 9_: 1-58.
Jenkins, D. and L.L. Medsker. 1964. Brucine Method for
Determination of Nitrate in Ocean, Estuarine, and Fresh
Waters. Anal. Chem. 36: 610-612.
Keeney, D.R. 1973. Nitrogen Cycle in Sediment - Water System.
J. Environ. Qual. 2_ (1): 15 p.
Kluesener, J.W. 1969. Oxygen and Color Relationship in
Petenwell Reservoir, Wisconsin River. M.S. Thesis.
University of Wisconsin, Madison. 116 p.
Kluesener, J. W. 1972. Nutrient Transport and Transforma-
tion in Lake Wingra, Wisconsin. Ph.D. Thesis. Univer-
sity of Wisconsin, Madison. 242 p.
Lee, G.F., M.T. Beatty, R.B. Corey, E.G. Fruh, C.L.R. Holt,
Jr., W.Hunter, G.W. Lawton, A.E. Peterson, F.H.
Schraufnagel, and K.B. Young. 1966. Report on the
Nutrient Source for Lake Mendota. Rep. to Lake Mendota
Problems Committee, Madison, Wisconsin. 56 p.
65
-------
Lonergan, D. Personal Communication. July, 1973.
Lopez, A.B. and N.L. Galvez. 1958. The Mineralization of
Organic Matter of Some Phillipine Soils Under Sub-
merged Conditions. Phillipine Agr. 42: 281-291.
Lueschow, L.A., J.M. Helm, D.R. Winter, and G.W. Karl.
Trophic Nature of Selected Wisconsin Lakes. Wise.
Acad. Sc., Arts and Letters. 58: 237-264.
Lund, J.W.G. 1965. The Ecology of Freshwater Phyto-
plankton. Cambridge Philosophical Society Biol.
Review. 40: 231-293.
Murray, S. 1971. Evaluation of Algal Assay Procedure--
PAAP Batch Test. Jour. WPCF. 4_3 (10): 1991-2003.
National Eutrophication Research Program. 1971. Algal
Assay Procedure: Bottle Test. Environmental Pro-
tection Agency, Corvallis, Oregon.
National Oceanographic and Atmospheric Administration.
1971. National Ocean Survey, Lake Survey Center:
Lake Ontario Chart No. 2. Ann Arbor, Michigan.
Nichols, D.S. and D.R. Keeney. 1973. Nitrogen and Phos-
phorus Release from Decaying Water Milfoil. Hydro-
biologia. 42: 509-525.
Orion Research, Inc. 1971. Instruction Manual for
Ammonia Electrode Model 95-10. Cambridge, Mass.
Painter, H.A. 1970. A Review of Literature on Inorganic
Nitrogen Metabolism in Microorganisms. Water
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Renn, C.E. 1937. Bacteria and the Phosphorus Cycle in the
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Sawyer, C.N., J.B. Lackey, and A.T. Lenz. 1944. Investi-
gation of the Odor Nuisance Occurring in the Madison
Lakes, Particularly Lakes Monona, Wanbesa, and
Kesonga from July 1943 to July 1944. Report to the
Governor's Commission, State of Wis. 92 p.
Sirisinha, K. 1973. Algal Available Nitrogen in Madison
Urban Storm Water Drainage and Selected Tributaries
of Lake Ontario. M.S. Thesis. University of Wis-
consin, Madison. 84 p.
Sonzogni, W. and G.F. Lee. 1972. Nutrient Sources for
Lake Mendota. Report to the Water Chemistry Program.
Univ. of Wise., Madison. 49 p.
66
-------
Tew, R.W. 1959. Laboratory Studies of Nitrogen Fixation
under Conditions Simulating Lake Environment. Ph.D.
Thesis. University of Wisconsin, Madison.
Vaccaro, R.F. 1965. Inorganic Nitrogen in Sea Water. Chem,
Oceanogr. !_: 365-408.
Von Brand, T., N.W. Rakestraw, and C.E. Renn. 1937. The
Experimental Decomposition and Regeneration of Nitro-
genous Organic Matter in Sea Water. Biol. Bull. 72:
165-175.
Waksman, S.A., C.L. Carey, and H.W. Reuszer. 1931. Marine
Bacteria and Their Role in the Cycle of Life in the
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Jour. WPCF. 36: 914-924.
67
-------
APPENDICES
Appendix Page
A Nitrogen Mineralization of New York River
Waters 591
Table A.I. - Nitrogen Mineralization of
New York River Waters 69
Table A.2. - Summary Data Table-Nitrogen
Mineralization of New York River Waters 76
Table A.3. - Mineralization Begun May 7,
1973-Genesee R. (42) and Oswego R. (4-3)
Particles in L. Ontario'Water from
Station 10 79
Table A.4. - Summary of Net Mean Nitrate-N
Released from Particles into No. 10 Lake
Water 80
Table A.5. - Mineralization Begun June 21,
1973-Genesee R. (58) and Oswego R. (59)
Particles in Lake Ontario Water from
Station 96 81
Table A. 6. - Summary of Net Mean Nitrate-N
Released from Particles into No. 96 Lake
Water 82
B Nitrogen Mineralization from Madison, Wis-
consin, Urban Runoff Samples 83
68
-------
APPENDIX A
Table A.I. NITROGEN MINERALIZATION OF NEW YORK RIVER WATERS
Sample No.
28
Incubation
Days
0
mgN/1 as:
Ammonia-N Nitrate-N
0.18 0.61
TKN
0.73
SKN
0.63
Oswego R.
March 2, 1973 10
25
50
82
31 0
Oswego R.
March 28, 1973 10
25
50
64
100
0.05
0.05
0.05
0.06
0.06
0.06
0.13
0.13
0.12
0.05
0.05
0.05
0.20
0.25
0.24
0.32
0.08
0.13
0.11
0.05
0.05
0.05
< 0.05
< 0.05
< 0.05
0.05
< 0.05
< 0.05
0.74
0.76
0.74
0.86
0.84
0.84
0.92
0.92
0.90
1.07
1.07
1.05
0.68
0.68
0.68
0.68
0.90
0.67
0.70
0.95
0.95
0.95
1.00
0.99
1.05
1.23
1.23
1.26
0.59 0.54
0.45
0.50
0.50
69
-------
Incubation mgN/1 as:
Sample No. Days Ammonia-N Nitrate-N TKN SKN
35 0 0.21 0.56 0.60 0.54
Oswego R.
April 7, 1973 10 0.35 0.68
0.32 0.68
0.32 0.68
25 0.10 0.92
0.10 0.92
0.10 0.85
50 0.05 0.92
0.05 1.05
0.05 1.05
64 0.05 1.05 0.47
0.06 1.05 0.60
0.06 1.05 0.55
100 < 0.05 1.20
< 0.05 1.65
0.06 1.26
43 0 0.28 0.46 0.96 0.43
Oswego R.
May 1, 1973 7
14
21
28
35
47 0 0.10 0.60 0.55 0.40
Oswego R.
May 14, 1973 10 0.05 0.60
0.05 0.68
0.07 0.60
0.24
0.25
0.28
0.10
0.11
0.10
0.10
0.12
0.11
0.06
0.07
0.06
0.08
0.07
0.07
0.46
0.46
0.46
0.80
0.80
0.82
0.82
0.82
0.82
0.95
0.90
0.92
0.87
0.90
0.87
0.85
0.71
1.13
0.57
0.65
—
0.61
0.61
0.48
0.36
0.36
0.45
0.50
0.35
0.50
0.41
0.41
0.45
70
-------
Sample No.
47
(continued)
48
Oswego R.
May 18, 1973
52
Oswego R.
May 28, 1973
59
Oswego R.
June 17, 1973
Incubation
Days
25
50
0
10
25
50
0
10
25
50
0
10
25
50
mgN/1
as:
Arnmonia-N Nitrate-N
0.09
0.06
0.06
—
—
—
0.29
0.23
0.19
0.21
0.20
0.17
0.20
< 0.05
< 0.05
0.05
0.31
0.37
0.22
0.30
0.22
0.17
0.09
< 0.05
0.06
< 0.05
0.58
0.34
0.31
0.25
< 0.05
< 0.05
< 0.05
—
—
—
0.88
0.72
—
0.82
0.79
0.80
0.25
0.68
0.62
0.57
—
—
—
0.74
0.77
0.79
0.35
0.62
0.68
0.66
0.88
0.88
0.88
0.90
1.01
0.92
0.68
0.75
0.74
0.82
1.37
1.29
1.36
1.25
1.22
1.24
TKN
SKN
0.76 0.49
1.14 0.55
1.62 1.17
71
-------
Incubation mgN/1 as:
Sample No. Days Ammonia-N Nitrate-N TKN SKN
34 0 0.15 0.98 1.23 0.52
Genesee R.
April 7, 1973 10 0.25 0.95
0.14 1.00
0.29 0.95
25 0.07 1.15
< 0.05 1.25
< 0.05 1.20
50 0.05 1.27
0.05 1.30
0.05 1.27
64 0.07 1.27 1.10
0.05 1.35 1.20
0.05 1.35 1.20
100 0.05 1.32
0.06 1.32
0.05 1.08
42 0 0.28 0.56 0.96 0.45
Genesee R.
May 1, 1973 7
14
21
28
35
51 0 0.32 0.35 0.91 0.47
Genesee R.
May 28, 1973 10 0.22 0.74
0.22 0.72
0.27 0.66
0.24
0.24
0.22
0.12
0.12
0.11
0.11
0.10
0.12
0.07
0.06
0.06
0.07
0.08
0.07
0.59
0.61
0.61
0.86
0.88
0.88
0.90
0.92
0.90
1.05
1.05
1.05
0.95
0.95
0.97
1.27
1.13
1.12
0.65
0.65
0.65
0.80
0.68
0.69
0.55
0.60
0.60
0.47
0.47
0.50
0.43
0.36
0.34
72
-------
Incubation mgN/1 as:
Sample No. Days Ammonia-N Nitrate-N TKN SKN
51 25 0.20 0.86
(continued) 0.05 0.85
< 0.05
50 0.06 0.92
< 0.05 0.88
< 0.05 0.91
58 0 0.60 0.64 1.62 1.04
Genesee R.
June 17, 1973 10 < 0.05 0.98
< 0.05 0.99
< 0.05 1.20
25 < 0.05 1.29
< 0.05 1.34
< 0.05
50 — 1.40
1.24
1.42
41 0 0.16 0.14 0.31 0.22
Niagara R.
April 30, 1973 7
14
21
28
35
50 0 0.15 0.36 0.46 0.33
Niagara R.
May 27, 1973 10 0.12 0.70
< 0.05 0.76
0.05 0.76
0.15
0.18
0.14
0.12
0.14
0.14
0.13
0.11
0.11
0.08
0.09
0.07
0.07
0.07
0.07
0.17
0.17
0.17
0.19
0.21
0.21
0.28
0.29
0.29
0.32
0.32
0.32
0.30
0.30
0.30
0.43
0.39
0.45
0.25
0.34
0.34
0.35
0.35
0.35
0.65
0.26
0.26
0.26
0.20
0.20
0.40
0.32
0.32
73
-------
Sample No.
50
(continued)
56
Niagara R.
June 16, 1973
36
Black R.
April 7, 1973
Incubation
Days
25
50
0
10
25
50
0
10
25
50
64
100
44
Black R.
May 1, 1973
mgN/1 as
:
Ammonia-N Nitrate-N
< 0.05
< 0.05
< 0.05
—
—
—
0.07
0.15
0.18
0.20
< 0.05
< 0.05
< 0.05
—
—
—
< 0.05
0.13
0.08
0.08
0.10
0.13
0.16
< 0.05
< 0.05
< 0.05
0.05
0.05
0.05
0.06
0.06
0.05
0.10
0.13
0.14
0.12
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.
0.
0.
0.
0.
0.
56
72
-
72
66
76
27
35
28
36
64
34
64
61
60
61
47
50
50
50
50
48
50
54
53
53
56
56
56
52
48
62
24
27
26
26
TKN
SKN
0.86 0.66
0.42 0.29
0.30
0.30
0.30
0.35 0.20
0.53
0.47
0.39
-------
Sample No.
44
(continued)
53
Black R.
May 28, 1973
60
Black R.
June 17, 1973
Incubation
Days
14
21
28
35
0
10
25
50
0
10
25
50
mgN/1 as:
Ammonia-N Nitrate-N
0.16
0.15
0.13
0.16
0.18
0.12
0.07
0.06
0.07
0.07
0.07
0.07
0.08
0.07
< 0.05
< 0.05
< 0.05
< 0.05
< 0.05
< 0.05
0.29
0.28
0.28
0.29
0.25
0.29
0.38
0.36
0.36
0.37
0.36
0.36
0.19
< 0
< 0
0
0
< 0
0
0
< 0
< 0
.05
.05
.07
.05
.05
.06
.05
.05
.05
0.28
0.64
0.28
0.56
0.39
0.56
0.40
0.35
0.35
0.27
0.22
0.16
0.16
0.42
0.45
0.38
0.36
0.49
0.50
TKN SKN
0.51
0.43
0.43
0.49
0.55
0.45
0.36
0.31
0.31
0.30
0.30
0.28
0.56 0.38
0.98 0.63
75
-------
Table A.2. SUMMARY DATA TABLE
NITROGEN MINERALIZATION OF NEW YORK RIVER WATERS
Sample Incubation NH
No.
28
Oswego
River
31
Oswego
River
35
Oswego
River
43
Oswego
River
47
Oswego
River
48
Oswego
River
Days
0
10
25
50
82
0
10
25
50
64
100
0
10
25
50
64
100
0
7
14
21
28
35
0
10
25
50
0
10
25
50
N
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.
3"
18
05
06
13
05
20
27
11
05
05
05
21
33
10
05
06
05
28
26
10
11
06
07
10
06
07
*
0.
0.
0.
< 0.
29
21
19
05
All Data: mgN/1
NO -
N
0
0
0
0
1
0
0
0
0
1
1
0
0
0
1
1
1
0
0
0
0
0
0
0
0
0
0
0
0
-
0
.61
.75
.85
.91
.06
.68
.68
.76
.95
.01
.24
.56
.68
.90
.01
.05
.37
.46
.46
.81
.82
.92
.88
.60
.63
.80
.80
.25
.62
.77
TKN SKN
0
0
0
0
0
0
0
0
0
0
0
0
0
0
.73 0.63
.22 0.15
.59 0.54
.48
.60 0.54
.54
.96 0.43
.90
.61
.57
.39
.45 0.42
.55 0.40
.76 0.49
NH
NO:
0
0
0
1
1
0
0
0
1
1
1
0
1
1
1
1
1
0
0
0
0
0
0
0
0
0
0
0
0
-
0
(A)
-N +
-N
.79
.80
.91
.04
.11
.88
.95
.87
.00
.01
.24
.77
.01
.00
.06
.11
.37
.74
.72
.91
.93
.98
.95
.70
.69
.87
.80
.54
.83
.77
(B)
Total (A)/(B)
N %
1.34 59
60
68
78
83
1.27 69
75
68
79
80
98
1.16 66
87
86
91
96
118
1.42 52
51
64
66
69
67
1.15 61
60
76
70
1.01 53
82
—
76
* Not determined; assumed equal to zero for sum of (nitrate+ammonia)-N
76
-------
Sample Incubation NH -
No.
52
Oswego
River
59
Oswego
River
34
Genesee
River
42
Genesee
River
51
Genesee
River
58
Genesee
River
41
Niagara
River
50
Niagara
River
Days
0
10
25
50
0
10
25
50
0
10
25
50
64
100
0
7
14
21
28
35
0
10
25
50
0
10
25
50
0
7
14
21
28
35
0
10
25
50
N
0.31
0.30
0.16
< 0.05
0.58
0.30
< 0.05
— *
0.15
0.23
< 0.05
0.05
0.06
0.05
0.28
0.23
0.12
0.11
0.06
0.07
0.32
0.24
0.08
< 0.05
0.60
< 0.05
< 0.05
__*
0.16
0.16
0.13
0.12
0.08
0.07
0.15
< 0.05
< 0.05
— *
NO -
N
0.35
0.65
0.88
0.94
0.68
0.77
1.34
1.24
0.98
0.97
1.20
1.28
1.32
1.24
0.56
0.60
0.87
0.91
1.05
0.96
0.35
0.71
0.86
0.90
0.64
1.06
1.32
1.35
0.14
0.17
0.20
0.29
0.32
0.30
0.36
0.74
0.64
0.71
TKN
1.14
1.62
1.23
1.17
0.96
1.17
0.65
0.72
0.58
0.48
0.91
1.62
0.31
0.42
0.31
0.35
0.26
0.22
0.46
(A)
NH -N +
SKN NO^-N
3
0.55 0.66
0.95
1.04
0.94
1.17 1.26
1.07
1.34
1.24
0.52 1.13
1.20
1.20
1.33
1.38
1.29
0.45 0.84
0.83
0.99
1.02
1.11
0.38 1.03
0.47 0.67
0.95
0.94
0.90
1.04 1.24
1.06
1.32
1.35
0.22 0.30
0.33
0.33
0.41
0.40
0.35 0.37
0.33 0.51
0.74
0.64
0.71
(B)
Total (A)/(B)
N %
1.49 44
64
70
63
2.30 55
46
58
54
2.21 51
55
54
60
63
59
1.52 55
55
65
67
73
68
1.26 53
75
75
71
2.26 55
47
58
60
0.45 67
73
73
91
89
82
0.82 62
90
78
87
-------
Sample Incubation NH -
No.
56
Niagara
River
36
Black
River
44
Black
River
53
Black
River
60
Black
River
Days
0
10
25
50
0
10
25
50
64
100
0
7
14
21
28
35
0
10
25
50
0
10
25
50
N
0.07
0.18
< 0.05
*
< 0.05
0.10
0.13
< 0.05
0.05
0.06
0.10
0.13
0.15
0.15
0.07
0.07
0.08
< 0.05
< 0.05
< 0.05
0.07
< 0.05
< 0.05
M_*
NO -
N
0.27
0.33
0.54
0.61
0.47
0.50
0.49
0.53
0.56
0.54
0.24
0.26
0.28
0.28
0.37
0.36
0.19
0.40
0.50
0.37
0.27
0.18
0.42
0.45
TKN
0.86
0.42
0.30
0.35
0.46
0.46
0.50
0.33
0.29
0.56
0.98
(A)
NH -N +
SKN NCL-N
0.66 0.34
0.51
0.54
0.61
0.29 0.47
0.60
0.62
0.53
0.61
0.60
0.20 0.34
0.39
0.43
0.43
0.44
0.43
0.38 0.27
0.40
0.50
0.37
0.63 0.34
0.18
0.42
0.45
(B)
Total (A)/(B)
N %
1.13 30
45
48
54
0.89 53
67
70
60
68
67
0.59 58
66
73
73
75
73
0.75 36
53
67
56
1.25 27
14
34
36
78
-------
Table A.3. MINERALIZATION BEGUN MAY 7, 1973
GENESEE R. (42) AND OSWEGO R. (43) PARTICLES IN L. ONTARIO
WATER FROM STATION 10
A. Initial Data
Sample No.
L. Ontario
water (10)
mean values
400 ml (10) +
Particles from
400 ml (42)
mean values
400 ml (10) +
Particles from
400 ml (43)
TKN (mgN/1)
0.24
0.24
0.24
0.25
0.54
0.54
0.68
0.59
0.36
0.36
0.50
NO~-N
j
0.
0.
0.
0.
0.
(mgN/1)
26
26
26
26
26
mean values 0.41 0.26
B. Dark Incubation (all values mgN/1 as NO -N)*
Time (days) 0_ 1 M. li !§.
Sample
L. Ontario
water (10)
mean values
400 ml (10) +
Particles from
400 ml (42)
mean values
400 ml (1.0) +
Particles from
400 ml (43)
mean values 0.26 0.32 0.38 0.39 0.37
0.26
0.26
0.26
0.26
__
—
—
0.26
__
—
—
0.26
0.26
0.26
0.26
0.29
0.29
0.29
0.29
0.32
0.32
0.33
0.27
0.27
0.27
0.27
0.35
0.34
0.35
0.35
0.38
0.38
0.38
0.27
0.27
0.27
0.27
0.34
0.34
0.34
0.34
0.40
0.38
0.40
0.25
0.27
0.26
0.26
0.35
0.35
0.35
0.35
0.36
0.36
0.38
* Each value shown represents the nitrate concentration of one test
bottle.
79
-------
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80
-------
Table A.S MINERALIZATION BEGUN JUNE 21, 1973
GENESEE R. (58) AND OSWEGO R. (59) PARTICLES IN LAKE ONTARIO
WATER FROM STATION 96
A. Initial Data
Sample No. TKN (mgN/1) NO^-N (mgN/1)
L. Ontario 0.49 0.0
water (96) 0.49 0.0
0.54 0.0
mean values 0.51 0.0
400 ml (96) + 1.10
Particles from 1.10
800 ml (58) 1.08
mean values 1.09 0.0
400 ml (96) + 0.96
Particles from 0.88
800 ml (59) 0.87
mean values 0.90 0.0
B. Dark Incubation (all values mgN/1 as NO -N)*
Time (days) 0_ 10 25 50
Sample No.
L. Ontario 0.0 0.0 0.0 0.18
water (96) 0.0 0.0 0.14 0.17
0.0 0.0 0.0 0.12
mean values 0.0 0.0 0.0 0.16
400 ml (96) + — 0.22 0.43 0.55
Particles from — 0.16 0.45 0.47
800 ml (58) — 0.16 0.40 0.51
mean values 0.0 0.18 0.43 0.51
400 ml (96) + — 0.0 0.43 0.52
Particles from -- 0.0 0.45 0.52
800 ml (59) — 0.0 0.51 0.56
mean values 0.0 0.0 0.46 0.53
* Each value shown represents the nitrate concentration of one test
bottle.
81
-------
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82
-------
APPENDIX B
NITROGEN MINERALIZATION FROM MADISON, WISCONSIN, URBAN RUNOFF SAMPLES
Sample
No.
A-6
A- 8
A- 9
Incubation
Day
0
10
25
50
100
0
10
25
50
100
0
10
NH+-N
(mgN/1)
0.13
0.67
0.74
0.69
0.15
0.20
0.20
0.05
0.05
0.05
0.16
0.16
0.04
0.59
0.59
0.59
0.61
0.91
0.80
0.87
0.21
0.21
0.16
0.08
0.08
0.08
0.80
1.35
1.58
1.35
NO~-N
(mgN/1)
0.02
0.08
0.05
0.05
0.93
1.06
0.98
1.15
1.20
1.20
1.62
1.56
1.56
0.45
0.52
0.53
0.50
0.48
0.50
0.49
1.50
1.50
1.47
1.75
1.75
1.90
0.21
0.40
0.40
0.45
Total
Kjeldahl-N
(mgN/1)
1.85
0.71
0.75
0.75
1.60
0.98
1.08
1.04
2.40
Soluble
Kjeldahl-N
(mgN/1)
1.00
0.32
0.32
0.32
0.67
0.45
0.45
0.38
1.70
83
-------
Sample
No.
A- 9
(cont'd)
A-12
B-6
Incubation
Day
25
50
100
0
10
25
50
82
0
10
25
. 50
100
NH'-N
4
(mgN/1)
0.26
0.30
0.29
0.30
0.34
0.26
0.07
0.08
0.05
1.12
0.43
1,78
1.98
1.71
1.62
1.71
1.74
1.44
1.72
0.10
0.12
0.10
0.14
1.09
1.14
1.21
0.21
0.10
0.13
0.05
0.05
0.22
0.14
0.23
0.19
NO -N
(mgN/1)
2.40
2.30
2.32
,60
,60
,50
,80
,80
2.75
0.71
0.93
0.97
0.97
,10
,12
,10
,32
,37
,32
3.75
3.75
3.75
0.005
0.02
0.03
0.02
48
48
41
1.95
1.75
1.85
2.85
2.85
2.10
Total
Kjeldahl-N
(mgN/1)
1.02
1.12
1.30
4.28
1.70
1.02
1.02
3.60
1.40
1.35
1.60
Soluble
Kjeldahl-N
(mgN/1)
0.66
0.66
0.66
2.40
0.52
0.52
0.20
1.20
0.63
0.63
0.55
B-8
0.75
0.62
1.50
0.75
-------
NH+-N NO'-N T°tal Soluble
Sample Incubation 4 3 Kjeldahl-N Kjeldahl-N
No. Day (mgN/1) (mgN/1) (mgN/1) (mgN/1)
B-8 10 0.70 0.72
(cont'd) 0.67 0.72
0.68 0.73
25 0.87 0.71
0.85 0.74
0.80 0.63
50 0.16 1.70
0.11 1.75
0.11 1.75
100 0.11 1.90 0.98 0.38
0.08 1.90 0.84 0.35
0.06 1.95 0.98 0.32
B-9 0 0.82 0.31 2.30 1.50
10
25
50
100
B-12 0 1.32 0.30 4.24 2.52
'lO 1.83 1.67
2.15 1.67
2.04 1.62
25 1.08 2.50
0.93 2.80
0.99 2.45
50 0.28 3.40
0.29 3.50
2.90
0.92
0.89
0.54
0.21
0.26
0.30
0.26
0.32
0.30
0.14
0.14
0.10
0.90
0.67
0.60
2.10
2.10
2.20
2.25
2.27
2.35
2.60 1.14
2.60 0.90
2.60 1.06
0.50
0.60
0.60
85
-------
Sample
No.
B-12
(cont'd)
D-6
D-8
D-10
Incubation
Day
82
0
10
25
50
100
0
10
25
50
100
0
10
25
4~N
(mgN/1)
0.10
0.11
0.10
0.08
0.78
0.65
0.64
0.05
0.18
0.05
0.05
0.12
0.15
0.16
0.12
0.12
0.64
0.59
0.43
0.49
0.76
0.60
0.51
0.11
0.21
0.21
0.06
0.06
0.06
0.51
0.46
0.46
0.39
0.21
0.30
0.26
NO -N
(mgN/1)
4.40
4.50
4.40
0.24
0.20
0.19
0.15
1.00
1.03
0.88
1.50
1.65
1.15
2.07
2.02
1.25
0.80
0.94
0.92
0.96
0.63
0.68
0.64
1.80
1.75
1.75
1.85
1.78
1.78
1.56
1.70
1.90
1.75
2.50
2.50
2.50
Total
Kjeldahl-N
(mgN/1)
1.40
1.60
1.80
2.45
1.70
1.55
1.50
1.70
1.08
1.22
1.16
2.70
Soluble
Kj eldahl-N
(mgN/1)
0.20
0.30
0.20
0.77
0.34
0.34
0.34
1.50
0.32
0.35
0.32
0.89
86
-------
NH+-N NO'-N T°tal S°1Uble
Sample Incubation 4 3 Kjeldahl-N Kjeldahl-N
No. Day (mgN/1) (mgN/1) (mgN/1) (mgN/1)
D-10 50 0.22 2.75
(cont'd) 0.26 2.68
0.24 2.68
100 0.08 3.00 2.24 0.50
0.10 3.00 2.30 0.56
0.08 2.90 2.30 0.56
D-12 0 1.07 0.33 4.60 2.52
10
25
50
100
F-9 0 0.85 0.63 1.60 1.20
10 0.92 0.80
1.16 0.75
1.20 0.70
25 0.21 2.00
0.30 1.98
0.32 1.80
50 0.15 2.22
0.17 2.22
0.20 1.95
100 0.10 2.10 0.62 0.56
0.08 2.35 0.56 0.50
0.13 2.40 0.74 0.50
2.00
1.25
1.04
1.05
-
1.10
0.54
0.50
0.49
0.08
0.10
0.10
1.90
1.95
1.90
2.05
2.05
2.02
2.90
3.00
3.00
3.60 2.00
3.60 2.08
3.70 2.08
0.20
0.40
0.20
87
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO. 2.
EPA-600/3-77-045
4. TITLE AND SUBTITLE ALGAL NunUM! AVAILABILITY AND LIMITA-
TION IN LAKE ONTARIO DURING IFYGL, PART II. Nitrogen
Available in Lake Ontario Tributary Water Samples and
Urban Runoff from Madison, Wisconsin
7. AUTHOR(S)
William F. Cowen*, Kannikar Sirisinha**, and
G. Fred Lee
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Center for Environmental Studies
University of Texas at Dallas
Richardson TX 75080
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Research Laboratory-Duluth MN
Office of Research and Development
U.S. Environmental Protection Agency
Duluth MN 55804
3. RECIPIENT'S ACCESSION-NO.
5. REPORT DATE
May 1977 issuing date
6. PERFORMING ORGANIZATION CODE
8. PERFORMING ORGANIZATION REPORT NO.
10. PROGRAM ELEMENT NO.
1BA608
11. CONTRACT/GRANT NO.
Contract R-800537-02
13. TYPE OF REPORT AND PERIOD COVERED
Final
14. SPONSORING AGENCY CODE
EPA/600/03
15. SUPPLEMENTARY NOTES *Medical-TechnicaHBioengineering Div., Edgewood Arsenal, Edgewood
MD 21040; **Faculuty of Public Health, Mahidol University, Rajvidhee Road, Bangkok,
Thailand
16. ABSTRACT
Samples of water from the Niagara, Genesee, Oswego and Black Rivers were collected
from March to June 1973. The samples were analyzed for nitrogen forms and were incu-
bated in darkness under aerobic conditions to promote mineralization of soluble
inorganic nitrogen from the organic nitrogen in the samples. The amounts of ammonia
and nitrate were determined as a function of the time of incubation. Generally over
50 percent of total nitrogen present in these river samples was immediately avail-
able for algal growth or potentially available after mineralization by bacteria.
The results were highly variable from each tributary, and no single value could be
selected from the data obtained to describe the availability of total nitrogen in
a given river.
U.S. Envirnri-nental Protection Agency
Region 5, Library (5K.-16)
230 3. Dearborn St-eet, Room 1670
Chicago, IL 6UC04
17. KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS
Nitrogen
Ammonia
Bacteria
Nutrients
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
b.lDENTIFIERS/OPEN ENDED TERMS
Lake Ontario
19. SECURITY CLASS (This Report)
TT^r.T.ASSTFTF.n
20. SECURITY CLASS (This page)
UNCLASSIFIED
c. COSATI Field/Group
06C, 06F
08H
21. NO. OP PAGES
100
22. PRICE
EPA Form 2220-1 (9-73)
U S. GOVERNMENT PRINTING OFFICE. 1977-757-056/6l|01 Region No. 5-II
------- |