c/EPA
            United States
            Environmental Protection
            Agency
            Environmental Research
            Laboratory
            Corvallis OR 97330
EPA 600 3-79-099
     '79
            Res
                  jpment
Effects of Forest
Fertilization with
Urea on Major
Biological
Components of
Small Cascade
Streams, Oregon


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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-600/3-79-099
                                             September 1979
EFFECTS OF FOREST FERTILIZATION WITH UREA ON MAJOR BIOLOGICAL
         COMPONENTS OF SMALL CASCADE STREAMS, OREGON
                             by
             F. S.  Stay, A.  Katko, K. W.  Malueg,
         M.  R.  Crouse, S.  E. Dominguez, R.  E.  Austin

                     Freshwater Division
         Corvallis Environmental Research Laboratory
                   Corvallis, Oregon 97330
                  This study was conducted
                     in cooperation with
                     U.S. Forest Service
                     Oakridge, OR 97492
         CORVALLIS ENVIRONMENTAL  RESEARCH  LABORATORY
             OFFICE OF  RESEARCH AND  DEVELOPMENT
            U.S.  ENVIRONMENTAL PROTECTION  AGENCY
                      CORVALLIS, OR 97330

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                                  DISCLAIMER

     This  report  has  been  reviewed by  the Corvallis  Environmental  Research
Laboratory, U.S.  Environmental  Protection  Agency,  and  approved  for publica-
tion.    Mention  of  trade names  or  commercial  products  does  not  constitute
endorsement or recommendation  for use.

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                                   FOREWORD

     Effective regulatory and enforcement actions by the Environmental  Protec-
tion Agency  would be  virtually  impossible without  sound scientific  data  on
pollutants  and  their   impact  on  environmental  stability  and human  health.
Responsibility for building  this  data base has  been  assigned  to  EPA's Office
of Research and Development and its 15 major field installations,  one of which
is the  Corvallis  Environmental Research Laboratory (CERL).

     The  primary mission of  the  Corvallis Laboratory is research  on  the ef-
fects  of  environmental  pollutants  on  terrestrial,  freshwater,   and marine
ecosystems; the behavior, effects and control of pollutants in lake and stream
systems;  and  the development  of  predictive models on the  movement of pollu-
tants in the biosphere.

     This  report evaluates the  impact of  forest  fertilization with  urea  on
major chemical and biological components of stream ecosystems.
                                                Thomas A.  Murphy
                                                Director,  CERL
                                      i i i

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                                   ABSTRACT

     During April, 1976, 1.9 x 103 ha of second growth Douglas fir, located in
the  Willamette  National  Forest  of  Oregon,  were  fertilized  with  224  kg
urea-N/ha.  Unfertilized  buffer  strips  of  60 and 90 m  were  maintained along
all  second and  third  order streams,  respectively.   Sharp increases  in  urea
concentrations (maximum of 12 mg/1) during the fertilization phase were due to
the  unintentional,  direct application to  the streams.   Immediately following
fertilization  all  nitrogen species  returned to  near  background levels.   The
Second  year  following  fertilization  only  N03-N02  appeared  to  be  slightly
elevated due to  fertilization.   Two-month fish bioassays using Salmo gairdneri
showed no mortalities which could be attributed to by-products or contaminants
of urea.  Algal  assays  using Selenastrum capricornutum, and chlorophyll a and
ATP-biomass of  periphyton  from  glass  slide  samplers  showed low supporting
capacity  and   generally no  significant  increase in  biomass resulting  from
fertilization.    Decomposition  experiments  using  dialysis  chambers  suggested
little difference between  a  fertilized and unfertilized station.  Analysis of
benthic  and drifting invertebrates suggested that little  change in community
structure could  be  related to  fertilization.  A record drought the first year
following  fertilization may have  resulted  in reduced  nitrogen loss  to  the
stream system.  This report covers  a period from April,  1976 to July,  1977 and
was completed  as  of June, 1979.

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                                   CONTENTS
Foreword	»	1]1
Abstract	   iv
Figures	   v^
Tables	vii
Acknowledgment	V111

     1.   Introduction	     1
     2.   Conclusions 	     3
     3.   Materials and Methods 	     5
               Site Description 	     5
               Fertilization	     5
               Physical Chemical Methods	     8
               Biological Methods 	     9
               Sample Collection Regimen	    12
     4.   Results and Discussion	    ^
               Precipitation and Streamflow 	    ^
               Stream Chemical Composition	    ^
               Fish Bioassays	    ^
               Algal Growth Potential 	    ^
               Periphyton Growth	    ^°
               Decomposition	.•	    ^
               Stream Benthic Communities and Invertebrate Drift	    41
                                                                            Cfi
References	     n

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                                    FIGURES

Number                                                                    Page

  1.  Map of  fertilized sub-areas, buffer zones, and sample stations ....  6

  2.  Opaque  plexiglass box and dialysis chamber used to measure
     decomposition	11

  3.  Flow at experimental and control stations before, during, and after
     fertilization of Huckleberry Flats 	 16

  4.  Urea-N  concentrations at experimental and control stream stations
     before, during, and after fertilization of Huckleberry Flats 	 20

  5.  Ammonia-N concentrations at experimental and control stream stations
     before, during and after fertilization of Huckleberry Flats  	 23

  6.  Nitrate-nitrite-N concentrations at experimental  and control stream
     stations before, during and after fertilization of Huckleberry Flats . 25

 7.  Total  Kjeldahl-N concentrations at experimental and control stream
     stations before, during, and after fertilization  of Huckleberry Flats  28

 8.  Urea-N loadings following fertilization in Huckleberry,  Fifth, Sixth,
     Seventh, Eighth and Ninth Creeks 	 31

 9.  Ammonia-N loadings  following fertilization in Huckleberry,  Fifth,
     Sixth,  Seventh, Eighth and Ninth Creeks	32

10.  Nitrate-nitrite loadings following fertilization  in Huckleberry,  Fifth,
     Sixth,  Seventh, Eighth and Ninth Creeks	33

11.  Algal  assay growth  yields for untreated streamwater; 1.0 mg/1
     nitrogen addition;  0.05 mg/1 phosphorus addition; micronutrient
     addition;  and 1.0 mg/1  nitrogen plus 0.05 ml/1  phosphorus addition .  . 34

12.  Chlorophyll  a and viable biomass concentrations of periphyton  on
     glass  slides~exposed in control and experimental  streams for 4 and 8
     week periods 	 37

13.  Numbers  of benthic  organisms in predominant orders at control  and
     experimental stations	42
                                     vi

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14.   Dendrogram developed from cluster analysis  of a  Bray-Curtis
     dissimilarity matrix of benthic invertebrates collected  from  control
     and experimental  stations before and after  fertilization of
     Huckleberry Flats	46

15.   Dendrogram developed from cluster analysis  of a  Bray-Curtis
     dissimilarity matrix of drifting invertebrates before  and during
     fertilization of  Huckleberry Flats 	 51

16.   Dendrogram developed from cluster analysis  of a  Bray-Curtis
     dissimilarity matrix of drifting invertebrates before, during and
     after fertilization of Huckleberry Flats 	 52


                                    TABLES

Number                                                                    Page

 1.   Collection, location and frequency of parameters analyzed for
     evaluation of forest fertilization at Huckleberry Flats	13

 2.   Precipitation at  Huckleberry Flats and a proximate NOAA  site  	 14

 3.   Average stream chemical composition in experimental and  control
     streams before, during, and after fertilization  of Huckleberry Flats . 18

 4.   Fish Bioassay; percent survival of rainbow trout, Sal mo  gairdneri,
     for a 2 month period during and after fertilization of Huckleberry
     Flats	35

 5.   Periphyton; mean  chlorophyll a and mean biomass  concentrations at all
     stations for each sample collection date and for all sample
     collection dates  at each station following fertilization of
     Huckleberry Flats	38

 6.   Heterotrophic Decomposition; percent decomposition and processing
     coefficients of Douglas fir, £. menziesii, and red alder, A.  rubra,
     substrates in Devils Canyon Creek and Seventh Creek	40

 7.   Total number of organisms, number of taxa, and Shannon-Wiener Diversity
     Index (SWDI) for invertebrate drift before,  during, and after
     fertilization of Huckleberry Flats  	48

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                               ACKNOWLEDGEMENTS

     The authors  are grateful  to  personnel of  the Oakridge District  of the
Willamette  National  Forest  for their  cooperation during  the  study,  to the
students of  Linn-Benton Community  College  who assisted  in the  sampling,  to
Peter  Klingeman  and  his staff  at  Oregon  State  University (OSU)  for their
assistance in gaging the streams, to Thomas Dudley and Peter Lattin of OSU who
assisted in the invertebrate enumeration and identification, to  Kenneth Harris
of CERL and  his  co-workers  for the chemical analyses conducted  in this study,
to Howard Mercier  and  William Faus of CERL for their computer programming and
contributions, and to  Charles Powers of CERL, Marvin  Allum of  CERL and Duane
Moore of the Pacific Northwest Forest and Range Experimental Station for their
technical  reviews.
                                     viii

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                                   SECTION 1

                                 INTRODUCTION

     A worldwide reduction in the land base available for silviculture  coupled
with  increasing  timber  demand have  stimulated the  development of improved
management methods to increase timber growth rate (Baule, 1975;  Schultz,  1975;
Haines,  1975;  Hansen,  1975).  Forest  fertilization  is one technique  for  in-
creasing timber yield which is gaining increased  acceptance.

     Groman (1972) and Baule (1975)  have reviewed the global  extent of forest
fertilization and predict a  rapid expansion of  its  use.  Baule1s review pre-
dicts that by 1980,  0.20-0.25% of the world's forest will have been  fertilized
at least  once.  While  this  is a relatively  modest fraction,  on a world scale
it will  involve the  use  of  a  prodigious  quantity  of fertilizer. And  it will
be concentrated in a  few, highly developed nations.

     A  rapid  increase  in forest  fertilization in  the  United  States  is  ex-
pected.  Fertilizers applied  include  N,  P, and K alone or in various combina-
tions.   In  the  Southeast United  States,  about  100,000 ha of  forest had been
fertilized  by 1973;  this is  expected  to  increase  greatly  (Haines,  1975).
Schultz  (1974) has estimated that in United States about 2 million ha/yr could
be fertilized.  In the  Pacific Northwest, about 250,000 ha of forest had been
fertilized with urea by  1974, with the annual  rate  approaching 120,000 ha/yr
(Moore,  1975a).  The combined fertilization potential in Oregon  and Washington
alone is estimated to be about one million  ha/yr  by 1985 (Hansen, 1974).

     Growth of  major coniferous  timber species  in  the  Pacific Northwest is
generally limited by nitrogen (Gessel, 1969).  Growth rate increases of up to
30%  have  been sustained  for 5-7 year periods by single applications of nitro-
gen  fertilizer  (Anderson,  1969).   In a review  of  nitrogen  dynamics in forest
soils,  Wollum and  Davey (1975)  predicted that modern  management  techniques
will place much higher demands on soil nitrogen  supplies because faster growth
rates  mean  shorter rotation  times and  higher nitrogen  removal  rates.  A doubl-
ing  of nitrogen demands is conceivable for conifers which store  a high propor-
tion of  their mature nitrogen content  early in life.   Logging  practices such
as whole-tree utilization and in-woods chipping will also result in increased
nitrogen removal.

     Because  of  the rapid  spread of  this  silvicultural practice,  there is
growing  concern  over  possible  adverse  side  effects  (Atkinson and Morison,
1975;  Safford,  1973).   Laboratory and field  studies on disposition of applied
urea nitrogen within forest soils have revealed little  likelihood of signifi-
cant nitrogen transport  into  groundwater (Cole  and Gessel,  1963;  1975; Cole
et a]_.  1975;  Knowles, 1975).  These  studies  have shown  that the ammonium ion,
the  hydrolysis product of urea,  is quickly immobilized by soil  cation exchange
sites  and biota.  Significant  losses  of fertilizer nitrogen have resulted from

                                       1

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ammonia  volatilization  (Watkins  et aJL  1972; Chin and Kroontje, 1963; Bernier
et  al.  1969;  Mahendrappa,  1975)  and  leaching to surface waters.  High volatil-
ization  losses have been noted under conditions of high pH which is due to the
greater  proportion of non-ionized ammonia.  Volatilization  losses  as high as
46% have been observed from forest soils in laboratory studies (Watkins et al.
1972).

      Leaching  of  fertilizer  nitrogen within  forest  soils  has  been observed
mainly  after  oxidation  of ammonium to nitrate.  Heilman (1974) found that the
nitrification  rate  in  Pacific  Northwestern  forest  soils  was  variable,  but
generally  low before and  immediately after fertilization.  However, nitrifi-
cation  was much  more  rapid in  both untreated and  re-fertilized soils taken
from  sites which had been fertilized some  years  previously.  It appears that
initial  fertilization  has a  "priming"  effect  on  nitrifying organisms, which
may lead to much higher rates of nitrate leaching with time and with repeated
urea treatments.

     Urea  fertilization  also  increases  nitrogen leaching through mobilization
of  organic N.   Salonius  (1972)  noted that application of urea increased the
rate  of decomposition  of  forest  humus,  and  Ogner  (1972a,b)  found greatly
increased  leaching  of  organic  N to  the  40  cm  depth after surface application
of  urea.

     Cole  and Gessel  (1974)   noted  that  three-fourths  of  the  usable water
supply  in  the United States originates in the forested one-third of the coun-
try.  As a result,  forest fertilization practices  have an inherent potential
to  affect  a large proportion  of the  nation's  water supply.   For this reason,
there have been many recent studies on streamwater contamination in fertilized
watersheds.

     Nitrogen  losses  to surface waters  represent a  very  small  fraction when
expressed  as  percent of  nitrogen applied; however, relatively high concentra-
tions have been  found.  Early  losses usually  range  from 2  to  3% of applied
nitrogen for  streams not  buffered by unfertilized  zones (Moore,  1975b)  and
about 0.5% for  streams with  buffer zones.   Such  losses may  be significant
(Werner, 1973) since they  impact headwater streams which average 1 km2 water-
shed per 1.4  km  of stream (Leopold et al_.  1964).  In watersheds fertilized at
the standard  rate of 224 kg N/ha, a  leaching  loss of only  1% per year trans-
lates to a  stream loading rate of 161 kg/km/yr.  Consequences of the impact of
this enrichment  upon the biota of streams and lakes have not been determined.
The only previous work  of which we  are aware involved  two streams draining
fertilized clearcuts in Alaska (Meehan et al_. 1975), which showed that changes
in  periphyton or benthic  biomass  due to fertilization could not be disting-
uished from natural variation.   Clearly,  more work is needed to ascertain bio-
logical  effects of forest fertilization on the large variety of stream systems
which may be affected.

     The present  study was undertaken in response to this  need.  The objective
of  this  study was to determine the impact of aerial forest fertilization with
urea on  major components of the aquatic ecosystem.  These components included
streamwater  chemical  composition,   fish  (bioassay),  algal   growth  potential
(algal assay), jji situ  periphyton growth, heterotrophic decomposition, benthic
community structure,  and  invertebrate drift composition.

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                                   SECTION 2

                                  CONCLUSIONS

     The chemical parameters,  other than the forms of  nitrogen,  demonstrated
no change  which could  be  attributed to  fertilization.  The  nitrogen  species
measured in this  study  included urea, NH3, NOs-NO^,  and total Kjeldahl  nitro-
gen.   Urea increased sharply during the fertilization period when unfertilized
buffer strips along  the streams were violated.   Due to the  possible  addition
of NH3 by red alder symbionts, very little could be concluded about fertiliza-
tion-ammonia  relationships.   Nitrate-nitrite-N, however,  did not  appear  af-
fected by  this  source  and increased slightly during  and  following fertiliza-
tion.   The year  following  fertilization,  the only  nitrogen  species  which
appeared to be elevated  due to fertilization was  N03-N02.

     The results  of this study suggest  that  fertilization slightly increased
stream nitrogen concentrations.   Streams  with higher flows and those that had
45 m  buffer strips  had lower  nitrogen concentrations  than streams with lower
flows and  30  m  strips.   Also, the  lack  of buffer  strips on the smaller head-
water tributaries resulted in elevated nitrogen concentrations even though the
remainder  of  the stream  was buffered.   Nitrogen  concentrations in an unbuf-
fered stream,  fertilized at  an application  rate  of about  one-third to two-
thirds  that used for  the other  watersheds,  were the  highest of all streams
studied.

     Nitrogen loadings  calculated  for  the experimental  streams were low and
possibly negative on occasion.  Higher N03-N02  loadings the second year appear
to be the  residual  effects of fertilization.  Although  nitrogen  concentrations
were  less   in comparable streams  buffered by  45  m  strips  (Station  11) than
those buffered  by 30 m strips  (Stations  23 and 24), loading  differences were
too small to be  measured.

     Two-month  fish bioassays showed no  increased fish mortality which could
be attributed to by-products  or contaminants of the urea fertilizer.

     Algal  assays  conducted on water from the  fertilized and control  streams
suggested  low growth potential  changed only slightly, if at  all,  after  fertil-
ization.   Fertilization did  not  shift   the  limiting  nutrient  status  of the
water  to  phosphorus.   The  experiments  did indicate that  nitrogen and phos-
phorus co-limited algal growth.

      The  periphyton studies  also  revealed  low growth potential.  However, a
slight  but significant (0.05) difference  in chlorophyll  a was  noted  between
control  and selected experimental  slides exposed  for 8 weeks.  Slides  exposed
for  4 weeks showed  no  significant difference,  and biomass  in the control and
experimental  stations  was not  significantly  different  for either 4  or  8  week

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exposures.  This  indicates  that fertilization may have  increased autotrophic
growth,  although  the  combined autotrophic-heterotrophic  supporting  capacity
did not change.

     The  heterotrophic decomposition  of Douglas  fir needles  and red  alder
leaves also suggested  a  low supporting capacity for  the  heterotrophic compo-
nent of  the stream biocoenosis.  Fertilization caused no measurable difference
in decomposition between the control and experimental  streams.

     Benthos  and  invertebrate  drift components of the aquatic  ecosystem also
showed  no change which could  be attributed  to  fertilization with urea.  The
changes  which did occur  in benthic community structure and invertebrate drift
composition appeared more  dependent upon emergence,  flow, and season than any
other factors.

     Violation of  buffer  zones resulted in temporary high nitrogen concentra-
tions, and the  lack  of any buffer  zone  along streams resulted in the highest
nitrogen  concentrations.   Also,  fertilization of  unbuffered  headwater tribu-
taries  appeared  to partially  nullify benefits gained  by buffering  the main
stream  channel.   Although  nitrogen  species  were sometimes slightly  elevated
following fertilization, other chemical  and biological parameters, except for
periphyton,  showed no change which could be attributed to fertilization.

     Although fertilization  appeared to  cause  a  slight  enhancement  of auto-
trophic growth,  the autotrophic-heterotrophic biomass supported by the streams
was  not altered.   Possible  changes in  supporting  capacity which  might have
occurred  in  response   to the  slightly increased nitrogen concentrations were
probably  mediated by  co-limitation of  phosphorus.   Had phosphorus  not been
co-limiting,  or had this  additional nitrogen been transported  to a nitrogen-
limited stream,  increased growth may have resulted.

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                                   SECTION 3

                             MATERIALS AND METHODS

SITE DESCRIPTION

     The study  site (Figure 1) was  a 1920 ha stand of  second-growth  Douglas
fir (Pseudotsuga menziesii) on  Huckleberry Flats  and adjacent areas  located  on
the west  slopes of  the  Cascade Mountains  in westcentral  Oregon.   This  stand
had been harvested  by  clearcutting in the  1930s.  Ground  vegetation was  pre-
dominantly  salal  Gaultheria sp_;  vine maple, Acer circinatum; Oregon  grape,
Berberis  sp;  and  Rhododendron sp.   Cottonwood,   Populus  sp,  and  red  alder,
Alnus rubra, were found in riparian areas.

     Huckleberry Flats  is classified  primarily  as land  type 14 (Legard  and
Meyer,  1973)  with  very  deep,  slightly plastic  to plastic  soils  derived from
residual and colluvial materials.   Surface soils  are thin sandy loams and silt
loams.  Subsoils  consist  of thick  silt  loams,  silty  clay loams, and  clay
loams.  Drainage is rapid in  the surface  soils  and moderate to slow  in the
subsoils.   In general the  land type on Huckleberry Flats is typical of flats,
benches, and terraces found in the Cascade Mountains of Oregon.

     Elevation of the  project  area ranges  from 800 m  to 1100 m.  The eastern
edge  has  a mean slope  of about  30%  and  the Flats area  has  a  mean slope of
about 10%.

     Prior  to the  selection  of areas to be fertilized, University of Washing-
ton  personnel  of  the Regional  Forest  Nutrition Research  Program initiated
studies in  1969 to  determine  the growth response of  Douglas  fir to nitrogen
fertilization in western Oregon  and Washington.  With  information developed
over  four growing  seasons (University of Washington,  1972;  1976),  the United
States Forest Service (USFS) identified Huckleberry Flats and Christy Flats as
two  major  areas to  be fertilized.   Huckleberry  Flats was  selected for this
study because it was large enough to  incorporate  the major portions of several
perennial first, second,  and third order  streams draining the watershed, and
had two streams  proximate to the  fertilized  area which could be used as con-
trols.

FERTILIZATION

     The fertilization of Huckleberry Flats was conducted  by the USFS, Willam-
ette  National  Forest,   Oakridge  District  during April,  1976.   Agricultural
grade urea  prills,  small pellets  1-2 mm  diameter, were applied to the  forest
with  a  gondola-spreader  carried by a helicopter.   The application rate was 224
kg N/ha.

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                                                                        on Devils Conyon Cr
                                                                        Roowvelt Cr) 2 Mi
                                                          FOREST FERTILIZATION PROJECT

                                                                Sampling Site
                                                                Limits and Date of Subareas
                                                                Fertilized
                                                                Approximate Drainage Area
                                                                Divide
                                                           119051
                                                           ^^*~  Road
Figure!.   Map  of  fertilized   sub-areas,  dates   of  urea  application,   buffer
            zones, and sample  stations.

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     During fertilization,  sub-areas  distinguishable  from  the air were fertil-
ized individually with known  amounts  of urea to  achieve the most accurate and
uniform application to the  total  project area.  Sub-areas, dates of  urea appli-
cations, buffer strips, and sampling station locations are shown in Fig. 1.

     Unfertilized buffer strips were  maintained  along  each side of the major
streams in the  fertilized  area.  Width  of the buffer strips was 45 m for the
two largest streams,  Huckleberry Creek  and Seventh Creek,  and  30 m for Fifth,
Sixth,  Salal, Eighth and Ninth Creeks.  Fertilization was  originally scheduled
to begin 1 April  1976 but  was postponed until 14  April because of heavy snow
cover.   Fertilization  was   intermittent  during 14-28 April  because  of heavy
rain and snow  storms.  Sampling  stations were located  on the  upper and lower
reaches of each  stream  in  the fertilized area except for  Eighth Creek, Ninth
Creek,   and Seventh  Creek  where  access was  not  possible.   The   only access
points   for Stations 2 and 3  were  located 0.3 km  inside the fertilized area,
and therefore could  not be used as upstream controls.

     Control streams  originally selected  for this  study  were Devils Canyon
Creek,   Station 25;  upper  Huckleberry  Creek, Station  15;  and Fourth Creek,
Stations 1 and  4.   Devils  Canyon Creek was selected  because  it was similar  in
size to Huckleberry Creek  and its  watershed was  completely separated  from the
fertilized area,  reducing  the possibility  of  contamination during fertiliza-
tion.

     Upper  Huckleberry  Creek,  Station  15,  was  considered representative  of
Huckleberry Creek,  Stations 8 and  13.  This station was  contaminated once  by
direct  fall during  fertilization;  however, because the area affected was  very
small,   and the  effects  short-term, Station 15 continued as a control.  Fourth
Creek,   Stations  1  and 4,  adjoins the  fertilized area and was selected as a
control because  of its similarity to the  smaller tributaries:   Fifth,  Sixth,
and Salal  Creeks.   Unfortunately,  during the fertilization of a plot south of
Huckleberry Flats on 13 April, the area between Road 1905 and some point below
Station 4  was  uniformly contaminated;  consequently, Station  4 could not  be
used as a control.   On  14 April,  Station  1 was  contaminated  and  the effects
persisted  for  an extended  period  of time.   Because of uniform contamination
above  Station  4, it  was used as an  example  of  an unbuffered  stream.  Visual
observations of  prill density  in  this area suggested  an  application rate  of
one-third  to  two-thirds that used in  the fertilized  area.   Because  of  this
lower rate of application,  the magnitude of effects found was considered equal
to  or  less than  that which  might have occurred  at the rate  of  224 kg  N/ha
applied to the  fertilized area.

     Although it was  not possible to obtain extensive antecedent characteriza-
tion of the relationship  between  the  control  and  the  experimental  streams,
comprehensive  characterization the month prior to the  treatment  suggested no
major differences in streamwater chemistry.

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 PHYSICAL-CHEMICAL METHODS

 Flow and Precipitation

      Flow measurements were made with  weirs  and staff gages at  each  station.
 Weirs were calibrated by  measuring  flows with a pigmy-gurly meter at  8 widely
 different stage  heights.  With this  information,  rating  functions were devel-
 oped to calculate flow  from  observed stage heights  at times of sample collec-
 tion.

      Precipitation was measured with 3  rain gages placed  on the higher eastern
 edge of  the  project area and with  5 gages in the  lower  Flats  area.   Because
 vandals destroyed  some  of the gages during  the study period, average values
 for the Flats  area  and  higher slopes were not always based on  the same number
 of samples.

 Chemical  Methods

      Water samples were analyzed in  the  field for conductivity,  pH, dissolved
 oxygen, and  alkalinity.   Subsamples  were  preserved with  4 mg HgCl2/l, 25  ml
 concentrated  HN03/1,  or  by cooling  to 4°C  (depending  on the analytical  re-
 quirements,  APHA, 1975),  and  shipped to  the Con/all is Environmental  Research
 Laboratory for  additional  chemical  analyses.    United  States  Environmental
 Protection Agency methods  (USEPA,  1974)  were used  for all  chemical  analyses
 except  urea.   Urea was hydrolyzed to ammonia by a modification of the Lienado
 and Rechnitz  (1974) method, and the  resultant ammonia was  analyzed by conven-
 tional  methods.  The  urea concentrations  were  calculated by subtracting  the
 ammonia-N  concentration  before hydrolysis  from  the ammonia-N  concentration
 after hydrolysis.

      Periphyton samples were analyzed for chlorophyll  a, phaeophytin,  and ATP.
 Chlorophyll  a  and phaeophytin were determined using a  modified  method  of
 Strickland and  Parsons  (1965) developed  by  Lorenzen (1967).  Absorbance  was
 measured  with  a dual  beam scanning  spectrophotometer at  750  and at 663  nm
 before  and after acidification with  two drops of IN  HC1.  Absorbance values  at
 663 nm  before  (663) and after  acidification (663  a)  were corrected for absorb-
 ance at 750 nm  and then  used  to  calculate chlorophyll a and phaeophytin with
 the equations  of Lorenzen.   Because  the  chlorophyll was  from  a periphyton
 community  these  equations were modified  to  present  results  as  weight/unit
 area:

        rhiA-nnhwii a  mn/m2 - 26.73  [(663)-(663  a)]-Extract  volume (L)
        Chlorophyll a, mg/m* - slide  Area (m58)-Cuvette Path  Length (cm)


        Phaormhu+i-n  mn/m2 - 26.73 [1.7 (663 a)-(663)]• Extract volume  (L)
        Phaeophytm, mg/m* -   glide  Area  (m^)-Cuvette Path  Length  (cm)

     Interferences resulting from a  shift  in the  fucoxanthin absorption spec-
trum  upon  over-acidification  have  been noted for this method (Riemann,  1978).
However,  inspection  of our sample  absorption spectra, and the lower  solvent
molarity used  (about 8  x 10-3 M HC1),  suggests  that interference from this
carotenoid did not occur.


                                      8

-------
     Samples for ATP analysis were extracted in the field within 2 hours  after
collection.   Samples were  first  filtered  through  a 0.45 urn membrane  filter,
then immediately extracted with  10 ml  of boiling  Tris  buffer,  pH 7.75 (Cheer
et  al_.  1974;  Holm-Hansen  and Booth,  1966).  The extract was frozen with  dry
ice and stored at -25°C until analyzed.  A refined luciferin-luciferase enzyme
was used  in the analysis.   Emitted light  was integrated for 1   minute with  a
commercial ATP photometer.  Purified disodium ATP standard was used to  prepare
calibration  curves   each  day ATP  analyses were  conducted.   Standards were
analyzed every  10 to 15 samples to detect  any change  in efficiency occurring
during the  period of  analysis.   ATP  concentration  in  the extracts was con-
verted to biomass  by assuming an  ATP  content of 2.4 mg ATP/g  dry  weight  or-
ganic matter (Weber, 1973).  This method estimates the  mass of viable organism
only and  values derived  by  this  method  are usually much  lower than  biomass
estimates derived from  gravimetric methods.

BIOLOGICAL METHODS

Fish Bioassay

     Acute toxicity experiments were conducted using fingerling  rainbow trout,
Sal mo gairdneri,  from  the Metolius Fish Hatchery, Camp Sherman, Oregon.  Fish
were placed in holding boxes,  at Station 4 on  Fourth  Creek, after adjusting
tank water  to  stream  temperature.  After a 24  hr acclimation  in the holding
boxes, 20  fingerlings about  8 to  10  cm long  were  placed in  each of 2 test
chambers  located  at  each  of  the  10 stations  selected for bioassays.  The fish
were then allowed to acclimate to the  individual  streams for  two weeks prior
to  fertilization.   They  were fed  trout  pellets  periodically  throughout  the
experiment.

Hacroi nvertebrates

     Three  individual  benthic samples  of 0.09 m2 were collected with  a  Surber
sampler  and composited (Hynes, 1970;  USGS,  1977;  USEPA, 1973).  The  sampling
location  at each  station was changed  each  trip to  avoid  areas previously
sampled.  Sample material  retained on  a #30 mesh  (0.5 mm)  sieve was preserved
in  buffered 10% formalin.   Invertebrates were sorted  from  the  debris by hand
picking, then preserved in 70% ethyl alcohol for counting and identification.

     Drifting  macroinvertebrates  were  collected with  0.30 m x 0.30 m x 1 m,
363 urn  mesh Nitex  nets.   When possible  the  entire  stream volume was  passed
through the nets.   Drifting  organisms  were  collected  for 24 hrs to eliminate
the effect  of diurnal  variations  in numbers  (Allan, 1978; Waters, 1972; Elliot,
1970).  Samples removed  from the nets were processed by the  same  procedures
used  for benthic  samples.   Organisms  were enumerated and  identified to the
lowest  taxonomic unit possible,  usually genus or  species.   Organisms which
could not be identified to  this  level  were considered a unique bit of  infor-
mation during the calculation of species diversity.

Algal Assays

     Algal  assays  were conducted according  to the Algal  Assay  Bottle  Test
(USEPA,  1971).  Water  samples were collected in  4  1  plastic bottles and stored

-------
at  4°C.   Samples were  autoclaved and  filtered,  then cultured  in triplicate
sets  as  follows:   streamwater without  nutrient  addition (control);  0.05  mg
P/l;  1.0 mg N/l; 1.0 mg N/l + 0.05 mg P/l; and a micronutrient spike, IX (AAM)
medium concentration.   Selenastrum capricornutum was used as the test alga.

Periphyton

      Periphyton  samples were  collected  from glass slides held  in a floating
acrylic plastic rack  (Sladeckova, 1962; Hohn and Hellerman, 1963).  Duplicate
sets  of  racks were placed at  each  station  for  4 and 8 week exposure periods.
Four  slides  from each  duplicate rack were  collected per exposure period and
duplicate  slides were  placed  in each  of  two wide-mouth bottles containing
streamwater.   Growth  on two  of the  four  slides  from  each  rack was removed,
filtered  with 0.45  pm membrane filters and extracted in boiling Tris for ATP.
Growth on the other  two  slides was analyzed for chlorophyll a.

Decomposition

      Decomposition  of red alder leaves and Douglas fir needles was measured in
duplicate  at  Stations  6  and 25 twice  during  the study.   Dialysis chambers
(McFeters and  Stuart,  1972)  with 8 |jm membrane pore size (Figure 2) were used
to  isolate  the  substrate  from the  stream  biota.   Both  red  alder leaves and
Douglas fir needles were collected from the study area during the fall season,
dried at  35°C,  and  ground  in  a Wiley  mill to pass through a #40  screen.  Known
amounts of  substrate  were  then sealed  in each dialysis chamber.  Before being
placed  in the stream,  each chamber was  inoculated with 100  ml of streamwater
taken with  a  50 ml syringe  from interstitial  spaces of the top  2  cm of the
bottom gravel.   It  was  assumed that the heterotrophic population contained in
this water and that which naturally occurred on the leaves would be of suffic-
ient  diversity  and density  to  initiate a  decomposer population  within the
chambers.

     The chambers were  placed in flow-through opaque plexiglas  boxes to pre-
vent  algal  growth  (Figure  2).  These  boxes  were then  completely submerged,
anchored to the  stream bottom, and aligned so that the stream flow would force
water  continuously through  the  boxes.   After two  months the  chambers were
removed  from  the   stream  and the  remaining residue collected.  Within two
hours, ATP  was  extracted  from duplicate subsamples  of the  residue.  The re-
mainder of the residue was stored at 4°C  until  another set of duplicate sub-
samples could  be filtered,  dried at  105°C,  and weighed.  This information was
used to determine total weight loss of the substrate and biomass of the heter-
otrophic population at time of collection.

     Leaching  experiments were also conducted on the same stock leaf material
used  in the  decomposition  study.   Two grams of leaf  substrate were placed in
200 ml  of distilled  water and  allowed to  stand at  about 20°C  for 24 hours.
Subsamples were  then filtered, dried at 105°C for 24 hr, and weighed to deter-
mine weight  loss due  to leaching.  Weight loss due to decomposition was esti-
mated by subtracting the estimated leaching loss from the total weight loss in
each  chamber.   Because  only  initial  and final weights could be  measured in
these experiments,  it was  assumed that weight loss conformed to the following
exponential  model (Petersen and Cummins, 1974):


                                     10

-------
Figure 2.   Opaque plexiglass box and dialysis chamber used to measure decompo-
           sition.


-------
                                 W  = W  e
                                  t    o

                        W.  = Weight remaining at time (t)
                        W  - Initial  weight
                        k  = Processing coefficient

Processing coefficients were calculated with the following equation:

                                     W,
SAMPLE COLLECTION REGIMEN
                               log  (rj)/t = -k
                                  e  W
     Sample collection  frequencies  and locations for all  parameters  measured
are presented  in Table 1, A  and  B.   Each  asterisk in Section B  represents  a
sample collection event at stations listed in Section A.
                                      12

-------
      TABLE 1.   COLLECTION,  LOCATION AND FREQUENCY OF PARAMETERS ANALYZED FOR EVALUATION OF FOREST FERTILIZATION AT HUCKLEBERRY  FLATS
      A  Location of Parameters  Sampled
      Parameters

        Chemistry & Streamflow
        Algal Assay
        Periphyton
        Macroinvertebrates
        Fish Bioassay
        Decomposition
                                          Stations  Sampled

                           1, 2, 3, 4,  6,  8,  9,  10,  11, 12, 13, 14, 15, 23, 24, 25
                           1,       4,  6,  8,  9,  10,  11, 12, 13,     15,         25
                           1,          6,  8,  9,      11, 12, 13,     15,         25
                           1,       4,  6,  8,  9,  10,  11, 12, 13,     15,         25
                           1,       4,  6,  8,  9,  10,  11, 12, 13, 14
                                       6,                                       25
B  Frequency of Parameters  Sampled

                         Feb       Mar
Parameters              	,   ,
Apr
                                                             Hay
 June
-4-H
                                                                          July
                                                                                    Auq
                                                           Sept
                                                                              Oct
Nov
Dec
CO
1976
  CTTemistry &
    Streamflow

  Algal Assay

  Periphyton

  Macroinvertebrates

  Fish Bioassay

  Decomposition
                                             * •* ^
                                             * '*•/',  *
                                             * •*,*   *   * *                           *
                                   *   *   **.**'****   *   **       ***   ***       *   *****
                                   *   * * *  * *'*.  *****************************
                                                                                                                      ****
                                                                                                                      ****
                                       * * *  ^X
                                              "***,
                                       *   *** •***
                                       * * *** •*>*'
                                                           * * * *   *   *  *
                                                           *********
1977
  Chemistry &
    Streamflow

  Periphyton

  Macroinvertebrates

  Decomposition
Feb
Mar

h
                                             Apr
                                                      May

                                                     H
                                                                       June
                              ****    *     *****   ****   **   *

                              *_	*	*	*	*
                                                                     *
      Fertilization per i od, 14-28 April.  1976. is indicated by the shaded area

-------
                                   SECTION 4

                            RESULTS AND DISCUSSION

PRECIPITATION AND STREAMFLOW

     Precipitation  data  for  Huckleberry  Flats,  adjacent  slopes, and  a  NOAA
weather  station  at  the  Oakridge  Salmon  Hatchery (USDC,  1976; 1977)  13  km
southeast  are  presented  in  Table  2.  Generally,  the  rainy season  for the
Pacific  Northwest begins about  October-November and  lasts  until early June.
After  June  very little  rainfall  occurs  until  autumn.   However during  the
1976-1977 winter, there was a severe Drought and precipitation was well below
normal.
   TABLE 2.  PRECIPITATION AT HUCKLEBERRY FLATS AND A PROXIMATE NOAA SITE.

Month
Jan. 1976
Feb.
Mar.
Apr.
May
June
July
Aug.
Sept.
Oct.
Nov.
Dec.
Jan. 1977
Feb.
Mar.
Apr.
May
June
Lower
Flats Area
(cm)
ND
ND
ND
1.7
4.6
0.6
4.3
4.3
1.1
1.0
0.0
ND
NO
2.7
22.3
5.3
19.6
2.0
Upper
Slopes Area
(cm)
ND
ND
ND
3.2
6.6
1.0
4.6
5.1
1.0
1.1
0.0
ND
ND
2.2
28.7
4.8
23.0
2.3
NOAA*
Site
(cm)
20.1
13.2
13.9
9.6
3.1
1.5
3.8
11.9
1.2
4.3
3.6
3.1
5.0
5.0
14.3
4.1
13.7
0.7
% of Monthly
Mean NOAA Site
(%)
117
107
112
116
49
26
317
900
35
62
23
17
28
40
117
50
216
12

* Oakridge Salmon Hatchery Located 13 km S.E. of Huckleberry Flats.
Elevation = Lower Flats Area ~  800 m
            Upper Slope Area ^ 1100 m
            Salmon Hatchery  ~  600 m
ND = no data available.
                                      14

-------
     Although precipitation at the  NOAA  site was quite different than  that  at
Huckleberry Flats, it  is  included  in Table 2 to  compare  the  1976-1977 winter
to an  average winter  derived  from a long-term  data  base.  During  September-
February 1976, when  precipitation  is usually highest, rainfall  was  29% of the
mean at the NOAA site and 76% of the mean for the total  study period.

     Although  extensive  antecedent  precipitation information at Huckleberry
Flats  is  not available  for comparison,  trends  were  similar to those at the
NOAA  site.   Rainfall  at  Huckleberry  Flats declined  during  the spring, in-
creased during July  and  August,  and then remained very  low  to  the  end of the
study except  for short periods in March and May.

     Stream  flow  (Figure 3) reflected the  extent of the drought at all  sta-
tions measured.  Flow  declined  from the  start of the  study  through the  1976-
1977 winter,  then  increased sharply.  Surface flow at Stations  2 and  10 com-
pletely stopped from late summer through winter.   Although there was generally
a  declining  hydrograph throughout  the  first year, flows did increase during
periods  of  higher precipitation.   Flows  were higher the second year during
March to June and peak values corresponded to increased rainfall.

     These data  suggest that  the  streams evaluated  for  this study fall  into
three  general groups.   Huckleberry  Creek,  Stations  8,  13,  15,  and Devils
Canyon  Creek consistently  exhibited the  highest flows  (102 to 103  I/sec).
Huckleberry  Creek  was  unique  because a very high proportion of its  flow orig-
inated  above  the fertilized portion  of the watershed.  Seventh Creek, Stations
6,  11;  Eighth Creek,  Station  24;  and Ninth Creek, Station  23;  had lower and
more variable  flows  (10° to 102 I/sec) than did Huckleberry Creek and Devils
Canyon  Creek.  Fourth,  Fifth,  Sixth and Sal a! Creeks generally had the lowest
flows ranging from 0  to 102 I/sec.

STREAM CHEMICAL COMPOSITIOON

     Average  chemical  composition  of the  streams  before,   during and after
fertilization  is  presented in Table  3.  The  "before"  average represents about
one month prior to fertilization and comprises the smallest number  of  samples.
The  "during"  average represents all  data collected during and two  weeks after
the  last  date of fertilization  in  the upstream  watershed.  These data include
the  effect  of direct  application  of urea  to the stream surface and  riparian
area.   The  "after"  average is derived from samples  collected  during the  re-
maining period of the study.

     In  general   the streams were  typical pristine  Cascade  mountain streams
(Fredriksen  et  a_K  1975) characterized by  soft  waters with  low  concentrations
of  dissolved materials.   Although the buffering capacity of these waters  was
low  as  indicated  by  the  alkalinity,  the  pH   varied  little throughout this
study.  Values  in  Table 3  were  determined_by first converting pH  to  hydrogen
ion  concentrations,  calculating the mean  (H) then converting this to pH.  Mean
values  of pH were similar  for  all  stations and  periods,  and  did not appear to
be affected by fertilization.
                                       15

-------
     10
    o
    LU
    to
     10
     10'
     iir1
        HBMJJ«SONOJFMBMJ


              1976            1977
                                              10
o »»'
UJ
in
                                              10
                                              10'
  Id-1
     HSMJJRSONOJFMSMJ

           1976            1977
                                                                                       10 '
o
LJ
LO
                                                                                      10 '
                                                                                      10'
                                                                                                         STATION 15
     MBMJJftSONOJFMflMJ

           1976            1977
     10 •
    o 10'
    LU
     10
CT1
     10'
     If1
                        STATION  6


                           FLOW
        MOMJJRSONDJFMftMJ


              1976            1977
                                              10
o 10
UJ
in
                                              10
-I 10°
                    STATION 11


                    "—• FLOW
     MftMJJOSONDJFMftMJ

           1976            1977
o
LU
LD
  10
                                                                                      10
                                                                                      10'
           1976
                    STATION 25


                       FLOU
                                                                                                                1977
    o '<>'
    LJ
    in
     iO
     10'
     KT1
                       STATION 23


                       •—• FLOU
                                              10
o
LU
in
                                              10
                                              10
  10-'
                    STATION 24


                    •—• PLOW
        MBMJJBSCNOJFH-4HJ         MRMJJASONDJFHftHJ

              1976            1977                    1976            1977

     Figure  3.   Flow at  experimental  and  control  stations before,  during,  and after fertilization  of Huckle-

                 berry Flats.

-------
o
Ul
  10
-I 10°
  10-'
                    STATION  9


                        FLOU
     MflMJJflSONDJFMBHJ


           1976             1977
                                                            STATION  2


                                                            •—• FLOU
                                         1IT1
                                            MfiMJJBSONOJFMflHJ

                                                  1976            1977
                                                                                 o
                                                                                 u
                                                                                    ID-
                                                                                 -1 10°
if1
                   STATION  1


                   •—• FLOW
   HftMJJASONDJFMAMJ

         197B            1977
  10
o
Ul
in
  10
  10'
                    STATION 10


                    •—• FLOW
                                           10 •
     MAMJJASONOJFMAHJ


           1976             1977
                                       o

                                       in
                                           10
                                           10'
                                           If"
                                                            STATION  3


                                                               FLOW
                                            MftMJJASONOJFMftMJ

                                                  1976            1977
                                                                                       MRMJjnSONOJFMAHJ
  10
o
Ul
  id
  10'
                    STATION 12


                    —• FLOW
                                           10 •
   HAMJJASONDJFMAMJ

         1976            1977


Figure  3.   (Continued)
                                       o
                                       LJ
                                       in
                                           10 •
                                           10'
                                           10'
                                                            STATION 14


                                                            •—• FLOW
                                              MAMJJASONOvlFNAMJ

                                                    1976            1977

-------
  TABLE 3.  AVERAGE STREAM CHEMICAL COMPOSITION IN EXPERIMENTAL AND CONTROL STREAMS BEFORE, CURING, AND AFTER FERTILIZATION OF HUCKLEBERRY FLATS
Devils Huckleberry Creek Salal Fourth Fifth Sixth Seventh Eighth Ninth
Creek Creek Creek Creek Creek Creek Creek Creek
Station #
Spec. Conductance
(umnos/cm)
Total Hardness
(ng CaC03/l)
Alkalinity (mg CaC03/l)
Total Inorganic -C
(rag C/l)
Total Organic -C
(mg C/l)
Total Kjeldahl -N
(rag N/l)
Ammonia (mg N/l)
Nitrate-Nitrite
(mg N/l)
Urea (rag N/l)
PH
Total Phosphate (mg P/l)
Orthoptiosphate (mg P/l)
Calcium (mg Ca/1)
Sodium (mg Na/1)
Potassium (mg K/l)
Magnesium (mg Mg/1)
Total Cation* (mg/1)

Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
Before
During
After
25 8 13 15 12 14 4 1 9 2 10 3 6 11 24 23
27.5 27.2 32.6 31.4 15.4 16.3 19.2 20.2 18.0 19.5 28.9 29.7 32.5 18.9 18.8 16.8
26.0 25.0 28.1 31.2 14.9 18.9 20.7 21.2 18.4 19.8 27.3 30.2 21.5 19.1 18.1 16.5
28.0 36.9 37.2 37.6 28.0 25.0 29.3 28.5 27.6 25.2 36.8 34.1 30.6 27.0 23.8 22.6
10.4 9.5 11.6 12.4 5.3 6.0 5.6 5.2 5.2 6.3 10.0 11.0 7.0 5.4 6.0 6.2
9.7 11.5 11.4 11.8 5.7 6.8 5.9 5.8 5.5 6.3 10.4 10.8 6.9 6.5 6.9 6.4
13.0 15.3 16.3 16.0 12.8 8.7 9.7 8.5 10.0 8.6 14.7 10.8 10.5 9.5 10.3 10.1
12.7 11.9 13.7 15.1 7.1 7.4 6.9 8.4 7.4 8.3 13.2 13.4 8.7 7.7 7.8 6.8
12.1 14.9 15.3 6.4 7.8 8.4 8.7 8.2 8.3 13.5 13.8 9.3 9.3
14.1 16.7 16.7 18.4 13.6 12.1 12.4 11.9 12.2 12.1 17.0 14.6 13.5 12.3 15.1 12.7
1.0 1.1 1.2 1.3 .9 .9 1.0 1.2 1.0 1.0 1.4 1.0 1.1 1.0 1.0 1.0
.7 .8 .9 .8 .8 .6 .7 .7 .8 .8 .9 1.1 .8 .8 .8 .8
1.0 1.0 1.1 1.2 1.2 1.0 1.1 1.0 1.0 1.0 1.1 1.0 1.0 1,0 1.0 1.0
.2 .3 .3 .2 .2 .2 .9 .8 .5 .2 .5 .5 .6 .2 .6 .9
.6 .5 .8 .6 .7 .5 .9 1.0 .7 .3 .9 .7 .5 .5 2.1 1.7
.7 .8 1.0 .8 .8 .6 1.9 1.6 1.5 .8 1.1 1.1 1.2 1.0 1.4 1.4
.087 .077 .047 .050 .054 .161 .100 .053 .050 .075 .063 .075 .067 .050 .050 .063
.063 .095 .472 2.379 .474 .704 .167 .086 .092 .205 .299 .296 .119 .298 .696 .056
.087 .066 .064 .065 .066 .077 .120 .091 .084 .084 .072 .085 .089 .077 .146 .080
.005 .005 .005 .005 .005 .005 .007 .006 .005 .007 .006 .005 .005 .005 .005 .005
.005 .005 .006 .005 .006 .007 .007 .005 .005 .007 .006 .011 .005 .010 .005 .005
.012 .006 .006 .006 .008 .010 .011 .008 .009 .007 .008 .008 .008 .008 .011 .009
.006 .005 .005 .005 .306 .006 .011 .013 .007 .005 .006 .005 .006 .005 .005 .005
.005 .007 .013 .005 .014 .010 .026 .009 .007 .008 .010 .007 .006 .007 .011 .005
.007 .008 .010 .006 .007 .020 .032 .015 ,009 .017 .011 .016 .008 .007 .019 .019
ND ND NO ND NO NO NO ND ND NO NO NO NO NO NO ND
.020 .059 .414 1.427 .565 .940 .091 .020 .047 .129 .347 .218 .066 .361 .187 .117
.020 .020 .020 .020 .020 .020 .033 .025 .023 .022 .025 .023 .020 .020 .024 .020
6.7 6.5 6.4 6.4 6.2 6.2 6.3 6.5 6.2 6.2 6.3 6.7 6.3 5.9 6.3 5.8
6.3 6.3 6.4 6.5 6.1 6.1 6.0 6.3 6.2 6.2 6.3 6.4 6.2 6.0 6.3 6.0
6.5 6.6 6.7 6.7 6.6 6.3 6.5 6.6 6.2 6.3 6.5 6.4 6.5 6.4 6.3 6.3
.014 .010 .015 .013 .011 .010 .010 .010 .010 .010 .010 .010 .010 .014 .012 .013
.010 .011 .010 .011 .010 .010 .012 .013 .011 .012 .011 .013 .013 .010 .010 .010
.012 .013 .011 .010 .011 .010 .011 .012 .014 .012 .011 .013 .016 .012 .011 .012
.008 .005 .005 .005 .005 .005 .005 .007 .005 .005 .006 .005 .005 .005 .005 .005
.005 .005 .005 .005 .005 .005 .005 .006 .005 .005 .006 .005 .005 .005 .005 .005
.006 .005 .005 .005 .005 .005 .005 .006 .005 .005 .005 .005 .005 .005 .005 .005
3.1 2.9 3.5 4.0 1.5 1.6 1.8 1.9 1.4 1.6 2.8 3.0 1.8 1.6 1.6 1.3
2.9 2.9 3.5 3.6 1.3 1.5 1.5 1.6 1.6 1.6 3.0 3.1 1.9 1.7 1.7 2.0
3.7 4.7 5.0 5.0 4.0 2.3 2.3 2.3 2.4 2.1 4.0 2.8 2.6 2.3 2.6 2.6
1.5 1.7 1.9 1.9 1.1 1.1 1.7 2.0 1.3 1.3 1.9 1.8 1.5 1.3 1.6 1.0
1.5 1 6 1.8 1.7 1.0 1.1 1.6 1.8 1.3 1.2 1.8 1.9 1.5 1.4 1.2 1.2
2.1 2.3 2.3 2.3 2.0 1.6 2.4 2.7 1.9 2.0 2.5 2.3 2.1 2.1 1.9 1.8
.2 .3 .2 .3 .2 .2 .5 .6 .4 .4 .4 .5 .4 .3 .3 .3
.2 .2 .2 .2 .1 .2 .4 .4 .3 .2 .4 .3 .3 .3 .3 .2
.3 .2 .2 .2 .2 .2 .4 .4 .3 .3 .3 .3 .3 .3 .3 .3
.6 .7 .6 .6 .5 .6 .6 .6 .6 .7 .8 .8 .7 .6 .5 .6
.6 .7 .6 .6 .5 .6 .5 .5 .7 .7 .9 .fi .7 .5 .6 .8
.9 8 8 8 .8 .8 .9 .7 .9 .9 1.1 .9 .9 .8 .8 .9
5.6 5.8 6.4 6.9 3.4 3.7 4.7 5.2 3.9 4.1 6.0 6.1 4.6 4.1 4.1 3.3
52 55 63 62 31 3.5 4.2 4.4 4.0 3.9 6.2 6.2 4.6 4.2 3.9 4.5
7.1 8.3 8.6 8.7 7.4 5.1 6.2 6.3 5.7 5.3 8.2 6.4 6.1 5.7 5.8 5.9
* Total Cation = The  sum  of  the Ca,  Na,  K,  and Mg  concentrations.

-------
     Average total cation concentrations  were  higher in the "after fertiliza-
tion" period in  the  controls  as well  as the experimental  stations, suggesting
that the  increase was  not  due to cation  exchange  caused  by fertilization,  as
noted by  others  (Aubertin  et  al.  1973; Cole and Gessel,  1963),  but was  prob-
ably due to other factors.

     Nutrient concentrations in the streams remained low throughout the study.
Average total phosphorus for  all  streams ranged between  the  analytical  limit
(0.010 mg P/l) and 0.016 mg P/l.  Orthophosphorus remained at or  near analyti-
cal limits at all stations with average values of 0.005-0.008 mg P/l. Average
urea and Kjeldahl nitrogen  (TKN) were higher during the fertilization phase  at
all experimental  stations,  when average urea-N and TKN ranges were 0.020-1.429
and  0.056-2.370  mg  N/l, respectively.   The  high  nitrogen  concentrations  at
Station 15 during fertilization were due to direct contamination  of the stream
at the time of one sample collection.  If the results of this sample are  omit-
ted, average urea-N  and  TKN are very  near  the  analytical limits of 0.022 and
0.100 mg/1,  respectively.

     Average nitrate-nitrite  and ammonia-nitrogen were at  or near analytical
limits during all three periods at both the control and experimental stations.
In medium to  low flow streams, average nitrate-nitrite  nitrogen was slightly
higher in the "during" and "after" fertilization periods than "before."

     Detailed information  on  urea, ammonia, nitrate-nitrite,  and  TKN is pre-
sented in Figures 5, 6, 7, and  8.

     Urea concentrations (Figure  4) were below analytical  limits  at all sta-
tions just  prior to  fertilization and from November  1976  through June  1977.
During fertilization all experimental  stations had short periods of increased
urea  concentration.   These high  values were  the  result of  inadvertant urea
application to the stream proper and proximate riparian areas.

     During fertilization,  two of the  control  streams,  upper Huckleberry and
Fourth Creeks, were  contaminated.   Because the impact at Station  15 was mini-
mal,  it  was retained  as a control.  Fourth Creek was  contaminated to  a much
greater extent and  could not  be used as a control, but was used as  an example
of  a stream  without  buffer  strips  because  urea  contamination  was uniform
throughout  its  watershed.   Also,  because the  application  rate  was only one-
third to two-thirds the 224 kg N/ha applied to  the fertilized  area,  the magni-
tude  of  effects was considered  equal  to or less than those which might have
occurred at the higher  application rate.

     Following  fertilization, urea  concentrations  at  Huckleberry Creek Sta-
tions  13  and  8 generally remained  below analytical limits.  However, Seventh
Creek Station  11, also  buffered by  45 m  strips,  showed extended periods of
increased  urea  concentrations  during  the post-fertilization  period.   This
difference  was  probably due  to a  combination  of  the direct fertilization of
the headwater streams  of Seventh Creek,  which  are mainly located  in the fer-
tilized area, and the greater proportion of flow in Huckleberry Creek origin-
ating outside  the fertilized area.  Because the  number  and extent of  buffer
zone violations were similar in both streams, as shown by urea peaks during
                                      19

-------
     10
     to'
    u
    O
     l(T3
STATION 8
•— • UREA
I
ff
I
'* I
IllP- Jl P



10°
or
UJ
h—
^JUT1
o
s: if2
If3






STATION 13
•— • UREA



Jl I *


10 -
10°
ct
UJ
(-

o
3





,

STATION 15
•— • UREft






        MftMJJASONDJFMOMJ
              1976            1977
                 MFIMJJOSONDJFMAMJ
                       1976           1977
                                       MflMJJdSONDJFMflMJ
                                             1976            1977
     10 •
     10'
    o:
    UJ
     ID'1

ro
o
              197B
                       STATION  6
                       •—• UREA
1977
                                             10
                                             10
             o:
             LU

                                             l(T
                                STATION 11
                                •—• UREA
                                                  t J»*M-fc-.
                                                                                    ID
                                                                                    10
                                   (T
                                   LJ

                                                                                    if
                                                      LIT3
                                                      STATION 25
                                                      *—• UREA
MftMJJflSONOJF  M«MJ
      1976            1977
MflMJJRSONDJFMAMJ
      1976            1977
     10
    tr

     ID'
     Id"3
                       STATION 23
                       •—• UREA

                                      JL
                                             10
                                             10
                                           o:
                                           LJ
                                             id"1
                                             ID"
              If3
                                STATION 24
                                •—• UREA
        H O H J ,J O S  0  N  0  J  F  M  ft  H  J
                                               MftMJJOSONOjFMFlMJ
              1976            1977                   1976            1977
     Figure 4.   Urea-N concentrations  at experimental  and  control  stream stations before,  during,  and after
                 fertilization of Huckleberry Flats.

-------
    10
  cr
    LIT1
  o
    1(T3
                     STATION 9


                     —• UREA
                  i .
                                           10
                                           L0
                                         cr
                                         UJ
                                         * ur2:
                                STATION  2


                                •—• UREA
                                                                                  10
                                                                                  10
                                                    cc
                                                    LJ
                                                                                O'
                                                STATION  1


                                                   UREA
       MflMJJflSONDJFMftMJ
                                              MflMJJfiSONDJFMflMJ
             1976
1977
1976
1977
MflMJJ«SONDJFMP»MJ

      1976            1977
    10
    10
  cr
ro
                     STATION 10


                     •—• UREA
                                           to
                                           10".
                                         cr
                                         UJ
                                           icr1
                                         o
                                           UT2
                                STATION  3


                                •—• UREA
                   .LJlA
                                                                                  10
                                                                                  10
                             CE
                             UJ


                             ^ICT1
                                                                                0
                                                                                  id"
                                                                                  !CT
                                                STATION  4


                                                   UREA
                                                                                                                 JLA,
       MBMJJfiSONOJFMPIMO
                                              MAMJJ(!|SONOJFMflMJ
             1976
1977
1976
1977
MAMJJASONDjFMflM.J

      1976            1977
    to
    10'
  cr
  LJ
    10-2
                      STATION 12


                      •—• UREA
         JJfl
             1976            1977

    Figure 4.  (Continued)
                                           10 •
                                           10'
             cr
             LJ

             « id"1
                                         o
                                           If3
                                STATION 14


                                •—• UREA
                 MSHJJFiSONDJFMBMJ

                       1976            1977

-------
 the  fertilization  period,  it is unlikely that these violations were responsi-
 ble for differences which occurred between these two streams.

      Urea  concentrations in streams buffered by 30 m unfertilized strips were
 generally  higher for  more  extended periods  than  were  Seventh or Huckleberry
 Creeks.  The  flows  at  Eighth and Ninth Creeks were similar to those at Station
 11  and were the highest of  the  streams  buffered by 30  m  strips.  Urea concen-
 trations  in these  two streams were  elevated for more extended periods than
 those at Station 11  and  were  less than those  in Fifth and Sixth  Creeks.  Lower
 Salal  Creek Station  12 had the lowest urea concentrations of streams with 30 m
 buffer strips.   Higher urea concentrations found  in streams  buffered  by 30 m
 strips appear to  be the result  of narrower buffer zones  and lower dilution
 and,  again,  not due  to the  number and  extent  of  buffer  zone violations.
 Fourth Creek  Station 4 had higher  urea  concentrations  for more extended peri-
 ods  than  all  other stations even  though  loadings were  less  than  224 kg/ha.
 This  was  'due  to the  absence  of buffer strips  to restrict  the transport of
 nitrogen to the streams (Fredriksen  et al. 1975).

      Ammonia  N increased at all  experimental stations  except 23 and 24 during
 the  fertilization  period (Figure 5).  Ammonia concentrations  in the controls
 generally remained  low during and following fertilization; however, concentra-
 tions  in Devils  Canyon Creek were  generally  higher  than  those in Huckleberry
 Creek.  These higher  values may  have  been  due  to  decomposition  of tissue
 sloughed from  the nitrogen-fixing symbionts associated  with riparian red alder
 (Moore, 1975a, b; Franklin et al. 1968) which were common near Station 25, but
 not  as  numerous  along  Huckleberry  Creek.   Urea  contamination  is  unlikely
 because the drainage  basin  of Devils Canyon Creek is well  removed from the
 fertilized  area.  Because of  this  additional ammonia  source and  its varia-
 bility among stations, no conclusions can be drawn about  the effect of fertil-
 ization on  ammonia  concentrations  in the experimental streams.  In general,
 concentrations were greatest at  all  stations during June-November 1976,  and
 increased slightly again  during May-June, 1977.

      Nitrate-nitrite  concentrations  increased  sharply  at  all  experimental
 stations during  the fertilization period (Figure 6), as  did urea,  apparently
 as  a  result  of buffer  zone violations.  In both control  stations,  N03-N02
 generally  remained  below analytical  limits  throughout the  study.   This sug-
 gests  that nitrogen  input from  the  red alder  did not  significantly affect
 N03-N02 concentrations,  and  the  increased concentrations found in the experi-
 mental streams were the result of fertilization.

     During  the  1976  post-fertilization  period,  N03-N02   concentrations  in
 Huckleberry  Creek  were  similar  to  those  in  the  controls  and  less than  all
 other  experimental stations.  Nitrate-nitrite concentrations in Seventh Creek,
which was  also buffered by 45 m strips, were greater than those in Huckleberry
 Cr.eek  and  less than most stations with  30 m  buffer zones.   Concentrations in
 streams buffered by 30  m  strips  were elevated for extended  periods  and were
 similar to  each  other.  Nitrate-nitrite concentrations were  highest  for more
extended periods in  Fourth Creek  than  in all  other  creeks  studied.   This,
again, is  a response to the  lack of unfertilized buffer zones  to  impede  the
transport of nitrogen to the stream.
                                      22

-------
     10
     10'
     If1
    0
     if2
     LIT3
                       STATION  8


                           AMMONIA
*
                                             10
                                            cr
                                            u
                                            C5
                                             i(T
                                              if-
                                              STATION 13


                                              •—• AMMONIA
                                                                                      10
                                                                  ct
                                                                  LJ
                                                                    If1
                                                                                    0
i(Tz
                                                                    HT3
                  STATION 15


                  •—• AMMONIA
        MflMJJflSONDJFMOMJ

              1976            1977
                               MflMJJASONDJFMOMJ

                                     1976            1977
   MflMJJOSONOJFMflMJ

         1976            1977
     IB1
    cr

no
CO
     if
     icr-
                       STATION  6


                       •—• AMMONIA
                                              ID
                                              10
                                            Ct
                                            LJ


                                            ^UT1
                                            3: id"2
                                              1(T-
                                              STATION 11


                                              •—• AMMONIA
                                                                                      10 '
                                                                    10 u



                                                                  LJ


                                                                  5*
                                                                  0
                                                                  2: if2
                                                                                      If3
                  STATION 25


                     AMMONIA
        MfiMJJftSONQjFMSMJ

              197B            1977
                               HAHJJASONDJFMAMJ

                                     1976            1977
   MflMJJOSONDJF  MBMJ

         1976            1977
     10
     10
    CE
    LJ
    o
      If'
                       STATION 23


                       •—• AMMONIA
                                              10
                                              10
                                            !*•
                                              If3
                                              STATION 24


                                              •—• AMMONIA
        MANJJftSONOJFMftMJ        HAMJJA50NDJFHANJ
               1976            1977                    1976            1977
     Figure 5.   Ammonia-N concentrations  at experimental and  control stream stations before, during  and after
                 fertilization of Huckleberry Flats.

-------
  10
  10°
tr
~ if1

  Iff 2
  l(T
                      STATION  9
                      •—• AMMONIA
                                              10
                                              10
                                            BE
                                            LJ
                                            0
                                              LIT
                      STATION  2
                      •—• AMMONIA
                                                                                A.
                                                                                         to
                                                                                         10
                                            tt
                                            LJ

     MflMJJftSONDJFMOMJ
            197B              1977
                                                                                         l(T
                                                                                         1CT
                                                                 STATION  1
                                                                 •—•  AMMONIA
     M«MJJ«SONDJFHflMJ
            1976              1977
                                                MAMJJOSONOJFMOMJ
                                                       1976             1977
tr
LJ
  *
O
  LIT
                      STATION 10
                      •—• AMMONIA
                                              10
CC
LJ
(—
-! if1
                                            0
  ID'3
                      STATION  3
                      •—• AMMONIA
     MAMJJRSONDJFMBMJ
            1976             1977
     HflMJJClSONDJFMfiMJ
            1976             1977
                                                                                         10
                                            cc
                                            LJ
                                                                                         ID'1
                                                                                       0
                                                                                         l(T3
                                                                 STATION  A
                                                                 •—• AMMONIA
                                                MflMJJflSONDJFMflMJ
                                                       1976             1977
  10
  10
tr
UJ
0
  1(T3
                     STATION 12
                     •—• AMMONIA

                                     A
                                             10
cr
UJ

  tr3
                     STATION 14
                     •—• AMMONIA
     MftMJJASONOJFMCiMJ
            1976             1977
     MfiMJjnSONDJFMflHJ
            1976             1977
 Figure 5.   (Continued)

-------

10°
tr
Ul

^
^>
T ifl-2
If3
to1
10°
PC
UJ
1-
^.
no
en
l(T3
10 S
10".
cc
Ul
3 Id"1
o
if3
STATION 8
•— • NITRATE+NITRITE

1
,
IP XU 11 /!


\L
MftMJJftSONDJFMftMJ
1976 1977
STATION 6
•— • NITRATE+NITRITE

-i^JllLL-VA
MRMJJflSONOJFMftMJ
1976 1977
STATION 23
— • NITRATE+NITRITE

h I
i flMjlk /I



-

10°
or
Ul
3 if1
o
10 l
10 «
tr
Ul
(-
X.
o
if3
10 »
10°
e
Ul
5 «r*
l(T3
STATION 13
•— • NITRATE+NITRITE

|
I.I II

MBMJJOSONOJFHOM.J
1976 1977
STATION 11
•— • NITRATE+NITRITE

J-JIlL 	 1
HfiHJJASONOJFHAMJ
1976 1977
STATION 24
•— • NITRATE+NITRITE
1
IL. t
_jJiA Ik. /i


10 °
a
Ul
"if1
0
Z If2
ir3
w5
10°
cc
Ul
H
_J
^
o
2 if
if^
STATION 15
•— • NITRATE+NITRITE


.1 A. 1 f\

MAMJjnSONOJFMAMJ
1976 1977
STATION 25
•— • NITRATE+NITRITE

L_U - All

HAMJJASONOJFMHMJ
1976 1977




    MAHJJASONOJFHAHJ
                                        MAMJJASONOJFMAMJ
         1976           1977                 1976           1977
Figure 6.   Nitrate-nitrite-N concentrations at experimental  and  control  stream stations before, during
           and after fertilization of Huckleberry  Flats.

-------
       10
       10
     tr
     LJ
     C5
                          STATION  9
                          •—• NITRATE+NITRITE
      if3!  ,  ,
                                                  to
                                                  10
                                            cr
                                            Q
                                            **
                                                O
                                                z
                                                  If
                                                                 STATION  2
                                                                 •—• NITRATE+NITRITE
     MAMJJASONDJFMAMJ
            1976             1977
                                                     MBMJJBSONDJFMflMJ
                                                           1976             1977
                                                                                                            STATION  1
                                                                                                            •—• NITRATE+NITRITE
                                                                                             iir
      10
cr
ui
**•
FN3   Z ,0-2
      MT3
                          STATION 10
                          •—• NITRATE+NITRITE
        I    ^»ll  llll
                                                  10
     MAMJJASONDJFMAMJ
            1976             1977
                                                  10
                                                tr
                                                Ul

                                             if3
                                                                 STATION  3
                                                                 •—• NITRATE+NITRITE
                                                     MAMJslASONOJFMBMJ
                                                           1976
                                                                        1977
                                                                                       cr
                                                                                       Ul
                                                                                             id"1
                                                                                             id"
                                                                                                            STATION  4
                                                                                                            •—• NITRATE+NITRITE
                                                                                           MflMJJftSONDJFMflMJ
                                                                                                  1976             1977
      10
cr
ui
Stri

                          STATION 12
                          •—• NITRATE+NITRITE
      tfl-3l  .  .
         MflMJJflSONOJFMAMJ
                1976             1977
    Figure 6.   (Continued)
                                                 10
                                                 10
                                                cr
                                                U
                                                Z ,8-2
                                             to-3
                                                                 STATION 14
                                                                 •—• NITRATE+NITRITE
                                                MAMJJASONDJFHAHJ
                                                       1976             1977

-------
     During the  second year,  N03-N02  was the predominant  inorganic  nitrogen
form in the stream.   Similar results have been found by others in the Pacific
Northwest (Moore,  1975b; Fredriksen et al_.  1975).

     Total  Kjeldahl  nitrogen  concentrations   in  Seventh Creek  and  all  30  m
buffered streams were similar to one another and greater than those in Huckle-
berry Creek (Figure 7).  Highest TKN concentrations occurred during fertiliza-
tion and  were  due to the increased  urea concentrations.  Following fertiliza-
tion, TKN  decreased  at all  experimental stations.  After August, it consisted
primarily  of  organic compounds other than  urea.   Differences  between concen-
trations  upstream and  downstream stations  as well  as  between control  and
experimental stations  were  not great enough to suggest the increased leaching
of organic nitrogen which  Ogner  (1972a;  1973) observed following fertiliza-
tion.

     These  data  suggest  that fertilization did increase the nitrogen content
in the  experimental  streams, and  that the 45  m unfertilized  buffer strips
combined  with  the  high  dilution  factor were effective  in  maintaining  low
nitrogen  concentrations  in   Huckleberry Creek.  The  combination  of unbuffered
headwater  streams  and lower  dilution  reduced the  effectiveness of  the 45  m
buffer strips  at  Seventh  Creek Station 11.   The  30  m buffer strips, although
not as effective  as  was  the 45 m  zone  for  Seventh Creek, were more effective
than no buffer strips, as  demonstrated at Fourth Creek.  Similar results have
been found  by  others  (Malueg et al_.  1972; Aubertin et al. 1973;  Moore, 1975b;
Thut, 1970).

     The  relatively  large   number  of  buffer  zone  violations  suggests  that
modifications   in  the  urea  application  procedure are needed to reduce unneces-
sary nutrient  loading to the stream systems.

     Urea, NH3 and  N03-N02  loadings for seven stream sections following fer-
tilization are presented  in Figures 8, 9 and  10, respectively.   The  loadings
presented  are  the  difference between the upstream  and  downstream station  and
are represent!ve  loadings for only that portion  of the watershed between the
two  stations.   Because many  of  the concentrations  used in calculating loads
were below analytical  limits,  upper and lower  limits of the  range which con-
tained the  actual  loading  value were defined  and presented in the figures as
two  separate  curves.   Therefore  when defining the  limits,  four  possible com-
binations could occur:

     Concentrations        Station Loadings        Watershed Loading  Range

     1)   Cj^A, C2£A     LX = Q1C1    L2 = Q2C2              D = QjCi  - Q2C2

     2)   Cj^A, C2£A   O^LiiQjA       L2 = Q2C2        -Q2C2SDSQiA -  Q2C2
                                      27

-------
    10
  cr
  LJ
    11T2
    if3
                  STATION 8
                     KJELDAHL N
v|lA_Uj_Jbl
JL
                                   U

                              1(T2
                              i(T3
                                             STATION 13


                                             •— • KJELDAHL N
                                          fl
-fcJLr^^—j*JlT\
           1976
                  1977
              1976
       1977
                                                                     10
                                                                     10
                                                             a
                                                             UJ
                                                                     if1
                                                                    o
                                       If2
                                                                     icr-
                                                      STATION 15


                                                      •— • KJELDAHL N
M  ft  M J  0  «


     1976
S  0 N 0  ,.)  f M


        1977
  cr
  LJ
  o
    itr2
CO
    itr3
                  STATION 6


                  •— • KJELDAHL N
       Nb^yA-----^^
                                   UJ

                              l(T3
                                             STATION 11


                                             •— • KJELDAHL N
                                                             cr
                                                             U)

                                                                      if-
                                                      STATION 25


                                                      •— • KJELDAHL M
      MAMJJOSONDJFMfiMJ


           1976         1977
                                MflMJJflSONOJFMOMJ

                                     1976          1977
                                          MBMJJfiSONDJFMflMJ


                                               1976          1977
  a
  UJ
  o
    if2
    If-
                   STATION 23


                   •—• KJELDAHL N
                                     to
                             cr
                             UJ
                             I If2
                                     If3
                                             STATION 24


                                               KJELDAHL N
                                       MflMJJBSONDjfMBMJ
           1976          1977                1976         1977

   Figure 7.  Total  Kjeldahl-N concentrations at experimental and  control  stream stations before,  during,

             and after fertilization of Huckleberry Flats.

-------
   a:
   LJ
     ID'1

     1CT3
                        STATION  9


                        •—• KJELDAHL N
              tr
              LJ
                                             CD
                icr3
                                   STATION 2


                                   •—• KJELOAHL  N
                                                                                        LJ
                                                          If1

                                                                                          if2
                                                          STATION  1


                                                          •—• KJELDAHL
        MSMJjaSONDJFMOMJ


              1976             1977
                   MflMJJOSOIVDJFMftMJ


                         197B            1977
                                          MSMJJASONDJFMflMJ


                                                 1976             1977
   cr
   LJ
     If1
ro
10
     iff3
                        STATION 10


                        •—• KJELDftHL N
              cc
              LJ

                                               icr2
                if3
                                   STATION 3


                                   •—• KJELDAHL N
                                      cr
                                      LJ
                                                                                        o
                                                                                          ur
                                                          STATION  4


                                                          •—• KJELDAHL N
        MflMJJfiSONOJFMfiHJ
              1976
1977
MftMJJflSONOJFMBMJ

       1976             1977
MftMJJfiSONDJFMOMJ

       1976             1977
     10-
   cr
   LJ
   •-; ur1
   o

1
\
STATION 12
•— • KJELDAHL N
K tflHi »i^ . 	 ^
10°
cr
LJ
"if1
•*v
x «r2
«
STATION 14
if •— • KJELDAHL N
JiuAuLA j\ Jm

               1976             1977

    Figure 7.   (Continued)

-------
           Where:

           Q!  and Q2 =  Flow at  the  lower and upper stations, respectively
           Cj  and C2 =  Concentration at lower and upper stations, respectively
           LI  and L2 =  Loadings (flow times concentrations) at lower and upper
                       stations,  respectively
                 D  =  Loadings (Lx - L2) for watershed between stations
                 A  =  Analytical limits; urea = 0.020 mg N/l; NH3 = 0.005
                       mg  N/l;  N03-N02 = 0.005 mg N/l.

      The  ranges  resulting from these calculations are the most accurate esti-
 mate  that can be made of the actual  loading values.

      Urea  loadings for  all  sections  ranged about  the  0 g  N/ha/day loading
 level  (Figure 8)  with a wider range at  higher flows.  No discernible differ-
 ences between streams buffered by 30 m  and 45 m  strips  were noted.  During
 1976,  urea loadings in Fourth  and  Salal Creeks were slightly higher than those
 in  Fifth  and  Sixth  Creeks.   During 1977,  ranges found  at  all  stations over-
 lapped and no differences  could be defined.

      Ammonia  also  generally  ranged about the 0 g N/ha/day loading level (Fig-
 ure 9).  Again, the  wider loading ranges found  in  Huckleberry Creek sections
 bracketed  ranges   found  at  all other  stations  and  no differences  could be
 defined.

      Nitrate-nitrite loading ranges in all  stream sections were higher in 1977
 than  in  1976  except  in Fourth  Creek (Figure  10).   The  highest loading values
 in Fourth Creek occurred during the spring of  1976.

      In general, these loading data suggest that urea and ammonia losses were
 low,  generally less  than 1  g/ha/day,  and  on  some  occasions  were  negative.
 Nitrate-nitrite loadings  in 1977  were  higher and more  variable  than in 1976
 probably  representing  the residual effects  of fertilization.   Differences in
 loading levels between streams buffered by 30 m, 45 m and 0 m strips were too
 small  to  be  measured.   The  negative N loading values found  for Fourth Creek,
 which  had no  buffer  zone,  were  due  to a  strong negative  hydraulic budget
 rather than decreased nitrogen  concentrations.

 FISH BIOASSAYS

      Fish  bioassays  using rainbow  trout,  Salmo gairdneri, were  conducted to
 determine possible fish mortality  due  to fertilization effects (Perna, 1971).
 The experiment was originally designed to compare survival  in a control stream,
 Fourth Creek, with survival  in  experimental streams.  Because Fourth Creek was
 contaminated,  no reference  can be made to an  untreated control.   Comparisons
between treated stations,  however, are possible and  informative.

     Percent  survival  of  the  trout  for a  2 month  period  during  and  after
fertilization  is presented   in  Table  4.   Generally,  survival was  high at all
stations.   Survival in the smaller streams  was very high,  90 to 100%, whereas
survival  in the largest  stream—Huckleberry  Creek at Station 8 and 13—was 78
and 82% respectively.

                                      30

-------
         STATION 8-12-13
                             UREA
     -

-------
     o
CO
IN3
         STATION 8-12-1
                             AMMONIA
                                           o
                                             -5
                                               STATION 13-15
                                                                   AMMONIA
         M«MJJ«50NDJF  MflMJ
               1976           1977
  MftMJJflSONDJFMBMJ
        1976            1977
                                               STATION  9-2
                                                                   AMMONIA
                                                                                     STATION 10-13
     o
                                                                                                          AMMONIA
5
4
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-------
            STATION B-12-13
       O
                                NITRATE+NITRITE     STATION  13-15
                                                                        NITRATES-NITRITE
                                               O
            MfiMJJOSONOJFMAH.i
                  1976            1977
                   MOMJJflSONOJFMHMJ
                         1976            1977
           STATION 6-9-10-11     NITRATE+NITRITE     STATION 9-2
CO
GO
5
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-------
                  I 12 >5 14 15
-
--
          0.25
         !
          0.25
          0.50r
          0.25
      -




      *   0.25



            10
      i a
      d '
      trO
           40
           20
                  I
                     »
 12  21  13  25
MAR MAR APR APR

28
JUN   AUG  SEP
                                                      9 10 1

                                                                     0.72
                                                                      I

                                                  12  21   13  25    28     4   21
                                                  MAR MAR APR APR   JUN   AUG SEP
                                                                       I 1  «
                                                                       •  a
                                                                       iWUk
                                                                                          in
 12  21 ' 13 ' 25    28     4   21
MAR MAR APR APR   AUG   JUN SEP
Figure 11.   Algal  assay growth yields  for  untreated streamwater;   1.0  mg/1 nitrogen addition; 0.05  mg/1
             phosphorus  addition;  micronutrient  addition;  and 1.0 mg/1  nitrogen plus 0.05 ml/1  phosphorus
             addition.

-------
 TABLE 4.   FISH BIOASSAY;  PERCENT SURVIVAL OF  RAINBOW  TROUT,  Sal mo gairdneri.
           FOR A 2 MONTH PERIOD DURING AND AFTER  FERTILIZATION OF HUCKLEBERRY
           FLATS


                   Station                 Percent  Survival*

                      1                           93
                      4                          93
                      9                          93
                     10                          98
                     11                           100
                     12                          90
                     14                          95
                      6                          95
                      8                          82t
                     13                          78ft

 * Exposure period was 13 April - 13 June, 1976.

 t 12.5 % Mortality before fertilization of upstream watershed.
tt 10.0 % Mortality before fertilization of upstream watershed.


     A  large  portion of  mortalities  at these two  stations  occurred  prior  to
fertilization of  their  upstream  watershed.  The  chemical  composition  of these
two stations was  affected the  least by fertilization, suggesting that factors
other  than  fertilization were  responsible  for the  higher mortality.   One
possible explanation  is stress caused by higher flow  velocities at  these two
stations.

     The results  of  these studies suggest that  percent mortality due to fer-
tilization-related processes was too low to be isolated from nonrelated causes
such as high  flow velocity.  These results are in  agreement with the  level  of
mortality  that  would be  expected considering the low ammonia  concentrations
and near neutral pH values found during the study  (USEPA, 1976).

ALGAL GROWTH POTENTIAL

     The algal  assay results  (Figure 11) show  the algal  growth potential  of
the stream  water  using  Selenastrum capricornutum under laboratory conditions.
These  data do  not  necessarily  reflect  a response  of the indigenous  algal
populations which may have been mediated by other  environmental  factors, e.g.
light  and   temperature.   They  do,  however,  represent the comparative growth
potential  of these samples with the test alga.

     Assays  of samples  without  nutrient spikes (untreated streamwater)  all
demonstrated very low growth yields (Miller et ah  1974; Maloney et al. 1972).
They showed no increase in yield after fertilization in comparison wrth the 12
March 1976 sampling prior to fertilization, or with the unfertilized controls,
Stations 15 and  25.  Only minor variations  in  yield throughout the  sampling
                                      35

-------
period were  observed,  and  significant  differences between stations were  not
apparent.

     The  addition  of a  combined micronutrient spike produced  no  significant
growth increase  over that of the controls.  This  indicated that a  micronutri-
ent  deficiency   did  not  exist,  and  would not mask  the  effect of  increased
nitrogen by limiting growth.

     Streamwater  from  nitrogen-poor  watersheds,  such as  those in the  study
area,  theoretically  would be  nitrogen  limited.   However,  the  nitrogen  addi-
tions  did not  indicate  nitrogen limitation prior to  fertilization  due  to  the
co-limiting  effect  of   low,  ambient  phosphorus  concentrations  (Table  3).
Orthophosphorus concentrations were  generally less than 5 ug/1.

     Another indication  of  an effect of increased nitrogen would  be  a  change
in  the nutrient limiting growth.   The  lack of growth response to  phosphorus
additions  suggests that  a shift to phosphorus limitation  did  not  occur imme-
diately  after fertilization.   At Stations  9,  10,  1, and  4, slight  phosphorus
limitation occurred  later in August, but  did  not  persist into September even
though  inorganic nitrogen and orthophosphorus concentrations  were  similar to
August values (Figure  11).   However,  the higher urea concentrations occurring
at  these  stations  on  4 October (Figure  4) would not be reflected in the inor-
ganic nitrogen values and may have been  the nitrogen source causing the  slight
shift to phosphorus limitation.

     Co-limitation  is  positively  indicated by  higher yields obtained  from
samples spiked with both nitrogen and phosphorus.   This  conclusion is  consis-
tant with the generally low concentrations of nitrogen and phosphorus found in
the streams during the  study period and  the general  lack  of response to  singu-
lar  nitrogen  or phosphorus  spikes.   Although  the yields were  sufficient to
eliminate primary  limitation  by  micronutrients,  the variation in yields among
the  stations  suggests that  micronutrients may have been  secondary  limiting
factors.

     These results strongly  indicate that fertilization  did  not increase  the
algal growth potential and,  in general, did not change  the limiting nutrient
status of  the stream water.   A slight shift to phosphorus limitation occurred
only on  one date in  the  streams which  generally  had the higher nitrogen con-
centrations (Figure 4).

PERIPHYTON GROWTH

     Assessment  of periphyton  growth was used to integrate possible  low rate
or  non-steady-state nutrient  loading which may or may not have been apparent
in the periodic grab samples for chemical analyses.

     Chlorophyll  a concentrations  and biomass as estimated by ATP concentra-
tions  in  periphyton collected from glass  slides which had been exposed for 4
and 8 week periods are presented in Figure 12.   In comparison  to other studies
presently being conducted in the Pacific Northwest, periphyton biomass was low
throughout the study period with maximum chlorophyll a and biomass values  of
                                      36

-------
CO
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1 6 8 9 II 12 131529

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1>T
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6 i 9 II 12 13 K 25
2 JUN 77
  Figure  12.   Chlorophyll  a  and  viable  biomass  concentrations  of periphyton  on  glass  slides  exposed in
               control  and  experimental  streams  for  4 and 8 week periods.

-------
2.5 and  178 mg/m2, respectively, for  the  4 week exposure period  and  8.5  and
70.8 mg/m2, respectively, for the 8 week exposure period.

     From  27  October 1976  to  2 March  1977,  growth at all  stations was very
low.  After this period growth  began to increase, and most stations reached a
maximum sometime during March to May.  During May to June, growth was probably
slowed by  the reduction in light resulting from leaf-out  and  development of
the riparian canopy.

     These  changes through  time are also shown by  the  mean chlorophyll a and
biomass concentrations of all  stations for each  4  and  8  week exposure period
presented  in  Table  5A.   The  variation among  stations  can  also be  seen by
comparing  means  of  all  samples  collected  during  the  study period for each
station (Table 5B).


 TABLE 5.   PERIPHYTON; MEAN  CHLOROPHYLL A AND MEAN BIOMASS CONCENTRATIONS AT
           ALL STATIONS FOR  EACH SAMPLE COLLECTION DATE AND FOR ALL SAMPLE
           COLLECTION DATES  AT EACH  STATION FOLLOWING FERTILIZATION OF HUCKLE-
           BERRY FLATS


A.   Means of all stations  for each  sampling date.
                      4 week exposure
8 week exposure
Date
27 Oct 1976
6 Dec 1976
2 Mar 1977
31 Mar 1977
4 May 1977
2 Jun 1977
Chlorophyll a
(mg/m2)
0.14
0.20
0.38
1.03
1.33
1.20
Biomass
(mg/m2)
4.1
6.8
-
48.4
60.2
24.2
Chlorophyll a
(mg/m2)

0.38

3.40

1.82
Biomass
(mg/m2)

12.5

76.3

48.2
B.    Means of all dates for each station
Station
1
6
8
9
11
12
13
15
25
Chlorophyll a
(mg/m2)
0.81
0.66
0.96
0.34
0.62
1.03
0.77
0.65
0.43
Biomass
(mg/m2)
6.5
10.5
26.0
4.4
24.6
7.7
41.0
40.0
55.1
Chlorophyll a
(mg/m2)
1.62
2.36
2.90
0.34
3.83
2.55
1.90
0.82
0.51
Biomass
(mg/m2)
32.3
11.5
12.1
41.0
50.8
36.3
86.5
84.5
56.0
                                      38

-------
     To determine if  a  significant difference (0.05) occurred between  fertil-
ized and non-fertilized stations, the sum of squares was partitioned  (Snedecor
and Cochran,  1967)  and  F-test  evaluated for experimental Stations 6, 8,  11,
and  13 and  control   Stations  15  and  25.   These  experimental stations were
selected on  the basis  of  their physical  similarity to  the  control  streams.
Differences  between the control  and fertilized stations  were  significant  for
chlorophyll  a  for the  8 week exposure  period.  Differences  in chlorophyll  a
for the 4  week period were not  significant.  When  comparing  the  mean  chloro-
phyll  a values  for  the 8 week exposure,  fertilized stations  consistently  had
higher chlorophyll a  values  than controls.  The only exception was  Station  9
which  was  located in an area heavily shaded with  a  dense  coniferous  canopy.
Biomass, which includes both autotrophs  and  heterotrophs,  was  not  signifi-
cantly  different  in  control  and experimental  streams  for  either the 4 or  8
week exposure periods.

     These  data suggest  that although  there  was  a slight  shift to a more
autotrophic  biomass,  as shown  by  chlorophyll a, the overall  biomass  was  not
changed by  fertilization.

DECOMPOSITION

     The results  of the decomposition experiments conducted at Stations 6  and
25 are summarized in  Table 6.  With the exception of Station 6, 1  April-2 June
1977  decomposition and processing  coefficients  for  Douglas  fir were  very
similar for the two test periods at both the control (25) and experimental  (6)
stations.   Decomposition and processing coefficients for Station 6,  1 April,
were about  one-half of those for  all  other Douglas fir  experiments and total
weight  loss  was less  than  estimated leaching  losses.  Biomass for these samp-
les,  however,   was  similar  to   the  other  samples  which indicates  these  low
values cannot  be  attributed to  a  high  decomposer  biomass.   A second analysis
was conducted  on  all  samples collected  on that date after having been frozen
for several  months.   These  results showed  Station  6  to be  similar to Station
25  and weight  losses  for  all  samples collected on that date  were  15 to  20%
higher after cold storage.   These  data  suggest that  the values presented for
Station 6,  1  April-2  June, are  low, and the true values  were probably similar
to  those  of Station  25.  However, because  the samples may have  changed,  the
data can only be used  for relative comparisons among  stations.

     Decomposition  and processing coefficients  for  red  alder were generally
higher  than  those for Douglas fir, probably  due to the  higher food quality  of
red alder  substrate (Petersen and  Cummins, 1974; Triska et  a]_.  1975).  Decomp-
osition of red alder was  marginally  but  consistently  higher  at the control
(25) than at the experimental  (6) station.

     Processing coefficients  for Douglas  fir  (0.0055 to  0.0064)  and red alder
(0.0072 to  0.0089)  measured  in  these  experiments  were  similar to those found
for  leaf packs (0.0049  to 0.0057  and  0.0077 to 0.0099,  respectively)  in three
replicate  experimental  Cascade  streams  by Triska  and Sedell (1976).  These
streams  uniquely  lacked an  invertebrate  leaf-shredder population  and  thus
closely  represented  microbial   decomposition processes  as  did  the  dialysis
chamber.  The investigators also  found  no discernible difference in leaf pack
                                       39

-------
 TABLE 6.  HETEROTROPHIC DECOMPOSITION; PERCENT DECOMPOSITION AND PROCESSING
           COEFFICIENTS OF DOUGLAS FIR, P. menziesii, AND RED ALDER, A. rubra,
           SUBSTRATES IN DEVILS CANYON CREEK AND SEVENTH CREEK


                 Incubation       Processingt  Decomposition?      Biomass
 Location           Period         Coefficient        (%)        (mg/g residue)
                                      (k)
Douglas fir Substrate
Seventh Cr.
Devils
Canyon Cr.
Seventh Cr.
Devils
Canyon Cr.
22 Aug-7 Dec 1976
22 Aug-7 Dec 1976
1 April -2 June 1977
1 April -2 June 1977
.0058
.0058
.0034
.0055
.0062
.0062
.0034
.0064
4 9
4 6
-10 -10
4 0
.103
.037
.057
.066
.086
.021
.178
.106
Alder Substrate
Seventh Cr.
Devils
Canyon Cr.
Seventh Cr.
Devils
Canyon Cr.
22 Aug-7 Dec 1976
22 Aug-7 Dec 1976
1 April -2 June 1977
1 April -2 June 1977
.0072
.0074
.0074
.0082
	
.0089
.0074
.0082
14 —
15 21
12 12
15 15
.139
.273
.033
.143
—
.089
.006
— — —

 t Conformance of decomposition to an experimental model was assumed and K was
calculated from ,     ,Wtv + _  .
                Loge W * ' "k
 I Decomposition equals total weight lost minus an estimated leaching loss of
29% for Douglas fir, and 25% for Red Alder.


decomposition in these three replicate streams which differed only in mean N03
concentrations (0.037,  0.059, and 0.138 mg N/l).

     In our  study,  the maximum range of  available  nitrogen (N03-N02 and NH3)
among all stations  was 0.010-0.038,  and for Stations 6 and 25 was 0.010-0.019
mg N/l.  These values  suggest  that no discernible difference in decomposition
would be expected among both control and  experimental  stations.   The  lack of
significant  differences  in  processing  coefficients at Stations 6  and  25 con-
firmed this  assumption.  Biomass  values,  as estimated from the ATP concentra-
tion at  the end of each experiment,  were all low,  ranging between  0.006 and

                                      40

-------
0.273 mg dry weight  biomass/g residue.   Although comparable biomass  data were
not available in  the  literature,  a range of reported ATP values was  converted
to biomass units  for  comparison.   The range of substrates  and  viable  biomass
included marine sediments at 130 mg/g (Ernst, 1970),  decomposing marine angio-
sperm disks at  22.8  mg/g (Knauer and Ayers,  1977),  and  mixed liquor volatile
solids from an activated sludge unit at 330 mg/g (Patterson et aL  1970).

     The decomposing marine  angiosperm,  although quite different,  is probably
the most similar  of  the three to the substrates used in these experiments and
is approximately  a factor of 80 greater than  our highest value.   An explana-
tion for these  low  biomass values may be that both  red alder and Douglas fir
substrates were initially nutrient limited and after leaching became  even more
so  (Triska et  a_K  1975).   This  condition  forces  the decomposers  to remove
nutrients, such as nitrogen  and phosphorus, from the  water phase  in order to
continue using  leaf  substrate  as  a  carbon  source  (Triska  and Sedell,  1976;
Triska  et  a]_.  1975;   Hynes  and Kaushik,  1969).   The low  decomposer  biomass
found in these chambers is in agreement with the nutrient-poor condition found
in the  water  phase  of these streams (Table 3) and the low supporting capacity
of these waters  for periphyton and S^ capricornutum in the algal assay.

     The  results  of  these  decomposition  experiments  suggest that  both the
control  and  experimental  streams  supported  a low  heterotrophic  biomass and
exhibited  low processing coefficients as a result of their oligotrophic condi-
tion.  These data also suggest that fertilization did not  cause a detectable
change in the  decomposer biomass or processing rate.

STREAM BENTHIC COMMUNITIES AND INVERTEBRATE DRIFT

     The numbers  of  benthic  invertebrates found  at  sampling stations and the
Shannon-Wiener  diversity  index  (SWDI) are presented  in Figure 13.  The predom-
inant organisms collected in the Surber samples  from all stations  throughout
the  study  belonged  to the orders Ephemeroptera  (mayflies),  Plecoptera (stone-
flies),  and  Trichoptera  (caddisflies).  Dipterans  (true flies) were  abundant
during  the spring at  Stations 6,  8,  10, and  11;  however, their numbers de-
creased  during the  summer.    Coleoptera (beetles),  Zygoptera  (damselflies),
Megoloptera  (alderflies and  dobsonflies),  and  Oligochaeta (segmented worms)
were  represented  by  only a  few  individuals.   A  variety  of  ephemeropterans and
Plecopterans  were  present  before  and  after fertilization.   Cinygmula sp,
Paraleptophlebia  sp,   Ironodes  nitidus,  and  Baetis  bicaudatus  were the most
commonly  encountered  ephemeropterans and exemplify  the variety of  the  mayfly
community.  The plecopterans were not as numerous nor as well represented  in
number  of taxa as  the mayflies.  The more  notable  were Alloperla  sp_,  Pelto-
perla  sp,  and Nemoura sp, which occurred both before  and after fertilization.
Trichopterans  were also collected  before  and after  fertilization  and were
fewer  in both numbers and kinds  than the  plecopterans.   The pattern of  occur-
rence  of  the  trichopterans  was  similar to that of  the ephemeropterans and
plecopterans  with Wormaldia SQ,  Micrasema  SJD,  and Rhyacophila sp_  occurring
most  often.   There  appeared  to  be  no consistent  pattern  or difference  in
species  composition  of the  benthic  communities sampled that could  be attrib-
uted to forest fertilization.
                                       41

-------
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600

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12 APR 21 APR 29 APR 3 MAY 5 AUG 28 OCT
1976
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                2 APR   13 APR   3 MAY   30 JUN   5 AUG   24 SEP   I APR
                1976    1976    1976    1976     1976    1976    1977

          SUOI         3.53

              SUOI *  SHANNON-UIENER DIVERSITY  INDEX
STATION 15

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1976 1976 1976
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          3.82

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400

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STATION 24


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2 APR  13 APR  3 MAY   30JUN   5 AUG   24 SEP   I APR
1976    1976    1976    1976     1976    1976    1977
       Figure 13.  Numbers of benthic  organisms in predominant orders  at control  and experimental  stations.

-------
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                SUDI s SHANNON-UIENER DIVERSITY  INDEX
Figure  13.  (Continued)

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-------
     The Shannon-Wiener diversity index (SWDI) was calculated only for samples
identified to  the lowest taxon  possible,  usually genus.  The  lowest indices
1.25 and 0.83 occurred on 12 April  1976 before fertilization  at Stations  6 and
11  respectively.  These  low values were due  to the extreme dominance of the
blackfly, Prosimuliurn  sg,  rather  than  a reduction  in  the  number  of species
present.  Generally,  the diversity index  values  for all  stations  before and
after  fertilization  ranged  between 0.83  and  4.48 with  no  trend which  would
suggest that  fertilization affected community diversity.

     In  an  attempt  to  discriminate  between   natural,  temporal, and spatial
variation in the  benthic community and those  variations  related to fertiliza-
tion,  a clustering  method  (Swartz,  1978; Boesch,  1977; Walker,  1974)  was
applied to a Bray-Curtis dissimilarity matrix developed for all  stations and
dates.   The results are presented as a dendrogram in Figure 14.

     If  fertilization had  a  measurable  effect  on  the  benthic  community  it
would be expected that samples collected before fertilization and from control
stations would  form different  clusters  than   stations affected by fertiliza-
tion, and clusters resulting from differences  in natural  stream habitats  would
appear separately for non-fertilized and fertilized stations.

     The dendrogram  shows  no  separation of fertilized and unfertilized  samp-
les.  Cluster I contains predominantly samples from streams with higher flows
and  from  stations at  higher  elevations with steeper slopes.   The  flows for
most of these  stations  at  the time  of  collection • were around  102  I/sec.
Within  this main  cluster there are three sub-clusters with higher intrafaunal
homogeneity.   The sub-cluster  farthest  to  the left contains samples collected
from the larger streams during the low flow summer-fall period  and from smaller
streams  during  the  spring  when  flows were  relatively  high.   The  other two
sub-clusters predominantly contained samples  collected during the spring from
larger streams.

     Samples clustered in Group  II were  mostly  collected during summer-fall
from stations  with  flows of  102  I/sec  or less.  Group III contains samp.les
collected from  mostly medium  to small  streams  during  the  spring when  flows
were relatively  high,  about  102  I/sec.  Samples  clustered  in  IV  were all
collected during  early  spring  when flows  were the  highest,  generally in the
102 to  103 I/sec range, from stations located in the Flats area.

     In general,  flow and season appeared to  be the factors most common to the
samples  contained within the  major clusters.  The  presence or  absence of a
fertilized upstream  watershed was  not a  common  factor  for any cluster.  Al-
though  Group IV had only one post-fertilization  sample, the samples  contained
within this cluster were all collected during early spring when  flows were the
highest.  All other  groups  contained samples  from both  control  and fertilized
stations or  from "before"  and "after"  fertilization  periods suggesting that
fertilization had no effect separable from natural causes.

     Drifting  invertebrates were  collected  from Salal,  Huckleberry, Fifth,
Sixth,   Seventh,  and  Fourth  Creeks.  As previously mentioned, Fourth  Creek was
originally intended  as  a  control  stream  but was heavily contaminated during
fertilization.   The predominant drift organisms collected in Fourth Creek were

                                      45

-------
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                         t — tnif>OJO — flDKOOtf><» — 0>0>	(0^OOtf> —*TIO —IOlOK>lf}K>CVOJ»OO»—IOK>€M	q-^OOtOOJOlO — CVJtO— rtlrtCJ — tflrt — fOlOCDf>tf»—— IftlOlO — C^WCSJffiKOtpPO^^i^i^^f' loioroioiov ooo''*'^^' ^^'f^^o^o^oco^rvoco^'^" OOK) uico^^'foto^'ioioo''- — - ___ _._, ,_._....__ ...._ .
MUlN I I  	LO—OOOOOO—— — OOOOOOOOOOOOOOOOOOOOOOOO—OOOOOOOOOO-OOOOOOOOOOOOOOOOO —OOOOOOOOOOOOOOOOOOOO
STATIC N —C  w  ^-_—	PJCJ	    
-------
Baetis bicaudatus and  Proslmulium  sp_.   Other organisms of significance in the
drift  included  the  ephemeropterans  Paraleptophlebia  s_£,  Ameletus  sp_,  and
Ironodes sp;  and  the plecopteran Nemoura sp.

     Organisms collected from  Fifth  and Sixth Creeks were  identified  only to
order; however,  dipterans  and  ephemeropterans were also  predominant  in these
drift samples.

     Seventh Creek was  also  predominated by Prosimulium s£ and B.  bicaudatus.
The plecopteran  Peltoperla s_p_ was prominant during early spring.  Other organ-
isms captured in significant numbers from Seventh Creek were the ephemeropter-
ans Paraleptophlebia sjp and Ameletus sjr, the plecopterans Nemoura sj>,  Isoperla
sj>  and  Pteronarcella sj);  and  the trichopterans Wormaldia  sp and  Rhyacophila
S£.

     Drift  in  lower  Huckleberry Creek was  dominated by  Prosimuiurn  s£,  B.
bicaudatus and Ironodes  m'tidus  whereas upper Huckleberry Creek was dominated
by  B.  bicaudatus.   Other  organisms  which were numerous  on  occasion  in upper
Huckleberry Creek were  !_.  nitidus,  Prosimulium sp, Nemoura sp_, Peltoperla sp,
Rhyacophila sp, and Micrasema sp.

     Drift in  Salal  Creek  differed from the other streams in that the plecop-
teran Nemoura sj) rather than B. bicaudatus shared dominance with the blackfly,
Prosimulium  sp.   Because of the  frequent occurrence  and  high  numbers of Ne-
moura sj)  in  Salal  Creek, this organism appeared  to be characteristic of this
stream.

     In  general, the  predominant organisms  drifting  in all  streams except
Salal Creek were the dipteran Prosimulium sj>, and the ephemeropteran B_. bicau-
datus.

     Total  numbers  of drifting organisms,  number  of  taxa, and Shannon-Weiner
diversity index  (SWDI) are presented in Table 7.  Drift samples collected  from
Stations  9  and  10 were  analyzed  to  order only and  therefore a diversity was
not calculated.  Highest drift occurred in early spring during  the pre-fertil-
ization and  fertilization phases when  flows were high.  Following this  period,
drift  decreased  as  did flow  (Figure  3).   Peak  drift  rates  which  occurred
during the  study were  usually due  to high numbers  of  Prosimulium S£ and B_._
bicaudatus.

     Index  values  for drift  in the  larger streams,  Seventh and Huckleberry
Creeks, were higher than those in the  smaller streams, Fourth and Salal  Creeks.
The lower values in these two smaller streams  were due  to  large numbers  of  a
few  species  drifting  and  low  evenness (Table 7),  rather than a reduction in
diversity of taxa present.

     These  data  suggest that  community  structure  with respect to diversity,
richness,  and types  of organisms present  was  independent of fertilization
dates  and  resultant  nitrogen  concentrations.   Variations  which  did  occur
appeared  more dependent on  season,   flow,  and possibly  emergence  period of
Prosimulium sp_ and B.  bicaudatus.
                                      47

-------
           TABLE 7.   TOTAL NUMBER OF ORGANISMS,  NUMBER OF TAXA,
                     FERTILIZATION OF HUCKLEBERRY FLATS
AND SHANNON-WIENER DIVERSITY INDEX (SWDI) FOR INVERTEBRATE  DRIFT BEFORE, DURING, AND AFTER
00
Station
Date
26 March
13 April
14 April
15 April
16 April
17 April
18 April
19 April
20 April
21 April
22 April
23 April
24 April
03 May
14 May
01 June
29 June
05 August
23 September
28 October
Total
350
255
605
388
360
150

99
148
207
86
124
162
30
160
128
193

34
39
Taxa
38
25
43
45
35
26

25
26
30
28
30

13
26
31
40



6
SWDI
3.46
3.11
3.75
3.96
3.55
3.59

3.47
3.77
3.76
4.36
4.37

3.21
3.60
4.42
4.43



Station 11
Total
1192
100



419
296
118
633
260
117
147
262

92
50
67



Taxa
28




29
32
25
39
28
26
13
37

14
8
21



SWDI
2.43




3.77
4.12
4.12
3.62
4.04
4.11
3.14
4.37

3.05
2.13
3.72



Station
Total
103
567
271
180

167
286
329
213
194

190
102

205
305
243
228
43

Taxa
27
40
30
27

24
31
32
25
32

28
28

20
37
36
34


8
SWDI
3.82
3.04
3.12
3.36

3.25
3.58
3.39
3.50
4.02

3.80
4.27

3.55
4.52
4.48
4.49


Station 13
Total Taxa SWDI

208 33 3.51
410 35 2.79
129 26 3.94
216 23 2.65
272 30 3.08
348 29 2.69
403 29 2.52
180 26 2.85
342 37 3.43




192 38 3.95



17
34
Station 9
Total

1438
1286
919
379
653
594
250
239
246

390
123

425
62

20
5
42
Station 10
Total Taxa SWDI
73
77 16 3.18
146
90 20 3.60
88

44


15
18
15
22

366
94
41



Station 12
Total

2972

1793
4042
2051
4682

1708
1891
1049
1069
753
763
1696
140
79
13


Taxa

20

24
26

16

27
24
24
23
32
13
20
9
17
7


SWDI

1.54

1.83
1.40

1.24

2.07
1.89
2.13
2.51
2.92
0.65
1.01
2.01
3.29
2.50


                                                                                                                                               (continued)

-------
TABLE 7 (cont.)
Station
Date
26 March
13 April
14 April
15 April
16 April
17 April
18 April
19 April
20 April
21 April
jg 22 April
23 April
24 April
03 May
14 May
01 June
29 June
Total

1716
2782
629

948
1778
980
2988
1108
2232
1078

32
533
26

14
Taxa SWDI


14
17

10
10
12
13
4
33
15

10
13
7



1.18
1.23

0.99
0.69
1.10
0.76
0.72
1.31
1.21

2.55
1.20
2.38

Station 1
Total Taxa SWOI
400 40 4. 28

658 28 1 . 59
530
458 27 1.61
595 28 3.18
441 28 1 . 78
42 13 2.77
121 27 3.91
905 36 2.07

286 24 2.15
515 32 1.88


54 21 3.60
111 26 3.48
Station 4
Total Taxa SWOI

74 12
587
191
311 27
151 22

84 25
66 25
95 24
189 28
220


69 18
375 30
90 22

3.05


3.55
3.64

3.99
4.11
3.86
3.89



3.07
3.17
3.80
05 August
23 September
28 October






12
71

29



-------
     Cluster analysis was conducted on these data.  Because the data base was
greater than the capacity of the computer program (Keniston, 1978), two matri-
cies were developed.  One included data from the "before" and "during" fertil-
ization periods (Figure 15), and the other included "before" and "after" as
well as every third day during the fertilization period (Figure 16).  The
dendrogram for the fertilization period (Figure 16) appears to have about five
main collection clusters with a high degree of dissimilarity and which contain
subgroups of high intra-group fauna!  homogeneity.

     Lower Huckleberry, lower Seventh and lower Fourth Creeks generally exhib-
ited a higher faunal homogeneity.  Upper Fourth Creek appears to be similar to
upper Huckleberry Creek, and Fifth Creek collection dates are separated into
two distinct periods, an early fertilization and a late fertilization period.
Salal Creek, as mentioned earlier, was unique and predominated Group V which
was also most dissimilar to all other groups.

     Within these subgroups of high faunal homogenity are stations with fer-
tilized and unfertilized watersheds, or pre-fertilization and post-fertiliza-
tion dates for the same stations, or both.

     In general the data suggest that drift populations in the Flats area are
dissimilar to those found on the steeper slopes, and that Salal Creek was
somewhat unique.   This analysis procedure, however, does not demonstrate any
grouping of fertilized or nonfertilized dates, suggesting that application of
the fertilizer had no measurable effects.

     The dendrogram for the combined fertilization and post-fertilization
periods is presented in Figure 16.   There are five main clusters which are
very dissimilar.   The subgroups formed within these main clusters generally
separate larger streams from smaller streams for spring, summer, and fall.
Due to the contamination of Fourth Creek (a control) few comparisons can be
made to stations with no unfertilized upstream watersheds.   The main comparison
which can be made is among stations which were exposed to different nitrogen
concentrations.   Stations on Huckleberry Creek generally had the lowest nitro-
gen concentrations and, following fertilization, were similar to the controls.
Seventh Creek and its tributaries were higher; and Fourth Creek Stations 1 and
4 had the highest nitrogen concentrations (Figures 4, 5, 6, and 7).

     In general,  stations with high nitrogen concentrations, as well as those
with low concentrations, were grouped together.   Also, each cluster contains
pre-fertilization dates.  Groupings,  however, appear to include similar size
streams or flows during similar seasons.   Cluster la is composed mainly of
summer Huckleberry Creek samples.  The samples from smaller streams contained
in this group were generally collected in spring when flows were higher.
Cluster Ib generally contained samples taken from the smaller streams during
early spring.   Cluster Ha consisted mainly of spring samples collected from
Seventh Creek whereas lib contained summer samples from both Seventh and
Fourth Creeks.   Cluster III lacked a high degree of intrafaunal homogeneity
and primarily contains Fourth Creek samples collected during early spring.
Samples from both large and small streams collected mainly during the summer
and fall  periods are contained in Cluster IV.  Salal Creek again appeared to
be unique and was the predominate station in Cluster V.


                                      50

-------
        l.0r
       0.9-
    YEAR
    DAY
    MONTH—[
    STATION-[
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                                                                                     *6=I976
Figure 15.   Dendrogram  developd from  cluster analysis of a  Bray-Curtis dissimilarity matrix of  drifting
             invertebrates before and during fertilization of  Huckleberry Flats.

-------
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     It appears that stream size, flow,  and season were factors  most common to
each cluster.  Although there were pre-fertilization samples contained in each
major  cluster,  it is  not  possible to  separate  the long-term  effects  of the
various  degrees  of  contamination and  stream size  on the drift  population.
However,  because  each  cluster  of high  faunal  homogeneity contained  samples
from streams which were exposed to both low  and  high  levels  of nitrogen con-
tamination,  as  well  as  samples  collected prior to  fertilization,  it appears
unlikely  that  fertilization had  a major effect on  invertebrate  drift  during
the study period.
                                       53

-------
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     Spectrophotometric Equations.   Limnol. and Oceanogr.  12:343-346.

Mahendrappa,  M.  K.   1975.   Ammonia  Volitilization  From  Some  Forest  Floor
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Maloney,  T.  E.,  W.   E.  Miller, and  T.  Shiroyama.   1972.  Algal   Responses  to
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                                      56

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                                       59

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                                   TECHNICAL REPORT DATA
                            (Please read Instructions on the reverse before completing)
1. REPORT NO.

 EPA-60Q/3-79-099
                                                           3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
 Effects of  Forest  Fertilization with Urea on Major
Biological Components  of Small  Cascade Streams, Oregon
                                                          5. REPORT DATE
                                                            September 1979
                                                          6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
 F. S. Stay, A.  Katko,  K.  W.  Malueg, M. R. Grouse,
 S. E. Dominguez,  R.  E.  Austin
                                                           8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
 Corvallis Environmental  Research Laboratory
 U. S. EPA Office  of Research and Development
 200 S. W. 35th  St.
 Con/all is, OR   97330
                                                          10. PROGRAM ELEMENT NO.

                                                             1BA820
                                                          11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
 same
                                                           13. TYPE OF REPORT AND PERIOD COVERED
                                                             inhouse.	1976-77
                                                           14. SPONSORING AGENCY CODE
                                                             EPA/600/02
15. SUPPLEMENTARY NOTES
 This study was conducted  in  cooperation with the U. S.  Forest  Service, Oakridge
 District Office, Oakridge, OR., 97492.
16. ABSTRACT
 During April,  1976,  1.9  x  10 ha of second growth Douglas  fir,  located in the Willa-
 mette National  Forest  of Oregon, were fertilized with 224 kg urea-N/ha.   Unfertilized
 buffer strips  of  60  and  90m were maintained along all second and  third order streams,
 respectively.   Sharp increases in urea concentrations (maximum of 12 mg/1) during
 the fertilization phase  were due to the unintentional, direct  application to the
 streams.  Immediately  following fertilization all nitrogen  species returned to near
 background levels.   The  second year following fertilization only  N03-N02 appeared to
 be slightly elevated due to fertilization.  Two-month fish  bioassays using Salmo
 gairdneri showed  no  mortalities which could be attributed to by-products or contami-
 nants of urea.  Algal  assays using Selenastrum capricornutum,  and chlorophyll a_ and
 ATP-biomass of periphyton  from glass slide samplers showed  low supporting capacity
 and generally  no  significant increase in biomass resulting  from fertilization.  Decom-
 position experiments using dialysis chambers suggested little  difference between a
 fertilized and unfertilized station.  Analysis of benthic and  drifting invertebrates
 suggested that little  change in community structure could be related to  fertilization.
 A record drought  the first year following fertilization may have  resulted in reduced
 nitrogen loss  to  the stream system.  This report covers a period  from April, 1976 to
 July, 1977 and was completed as of June, 1979.
17.
                               KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.lDENTIFIERS/OPEN ENDED TERMS
                                                                        c.  COSATI Field/Croup
                                                                         Biological/Envir-
                                                                         onmental  Biology
                                                                         =6F
                                                                         Chemistry/Inorgan
                                                                         ic Chemistry =7B
Urea, Fertilizer, Nitrate,  Ammonia, Nutri-
ents, Invertebrate, Algae,  Potential,
Periphyton, Decomposition,  Benthos, Fish
18. DISTRIBUTION STATEMENT
Release Unlimited
                                             19. SECURITY CLASS (ThisReport)
                                               unclassified
                                                                         21. NO. OF PAGES
                                              20. SECURITY CLASS (Thispage)
                                               unclassified
                                                                        22. PRICE
EPA Form 2220-1 (Rev. 4-77)
                     PREVIOUS EDITION IS OBSOLETt.
                                            60

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