A STUDY OF NUTREENT ACCRUAL, UPTAKE,
AND REGENERATRON A5 RELATED TO

PRIMARY FRODUC‘FION IN A
WARM-WAKE SffiEAM

Thesis for H19 Degree of Dh. D.
MICHIGAN STATE UNIVERSITY

Morris Leroy Brehmer
1958

WWW m1mnmmmrrmmm

3 1293 00658 5636

This is to certify that the 4- '23
0' ' ‘-

thesis entitled ",

A STUDY OF NUTRIENT ACCRUAL, UPTAKE, AND REGENERATION
AS RELATED TO PRIMARY PRODUCTION IN A WARM-WATER STREAM

presented by

MORRIS LEROY BREWER

has been accepted towards fulfillment

of the requirements for

PhoDo degree in FiSheI'iQS 8C Wildlife

(Mm Q Ml

Major professor

Date N0 b 0

 

 

  
      

  

L [B R A R Y
Michigan State
University

 

A STUDY OF NUTRIENT ACCRUAL, UPTAKE, AND REGENERATION

AS RELATED TO PRIMARY PRODUCTION IN A NARM¥WATER STREAM

HORRIS LEROY BREHMER

AN’ABSTRACT

submitted to the School for Advanced Graduate Studies of
Michigan State University of Agriculture and
Applied Science in partial fulfillment of

the requirements for the degree of
DOCTOR OF PHILOSOPHY

Department of Fisheries and Wildlife
1958

. Approved Rikki} § (8%»ng

Morris Leroy Brehmer

.The Red Cedar River, a'warmdwater stream which drains a portion
of the south-central part of the lower peninsula of Michigan, was in-
vestigated to determine the major nutrient sources and the relationShip
between primary production and nutrient levels.

The results of a study of the entire stream indicated that the
major changes in nutrient levels occurred after the introduction of
effluents from municipal drains and sewage treatment plant outfalls. The
nutrient contributions from the tributary streams were of the greatest
magnitude during periods of high run-off from the watershed area and
had little effect on the nutrient budget of the main stream.

An intensive study was made on a 5.3 mile area of the river to
determine the rate of nutrient accrual, uptake, and regeneration as
related to the seasonal primary production patterns. Upstream from
this area a reservoir served as a silt basin, just within the upper
limits a turbid tributary stream emptied into the river, and within
the first 0.5 mile the outfall from a sewage treatment plant provided
a continuous source of nutrients without producing septic conditions.

The dissolved, sestonic, acidssoluble sestonic, and total phosphorus,
the ammonia and nitrite plus nitrate nitrogen, and the periphyton
production were determined at nine stations within the 5.3 mile area
during a 13-month period. The periphyton measurements involved the
use of plexiglass artificial substrata. The Optical absorbency of
the ethanol-extracted phytopigments from the periphyton accumulation

on the substrata was used as an index of production.

 

Morris Leroy Brehmer

The data indicated that during average water-level conditions the
stream was enriched by more than 100 ug I“ of phosphorus and 0.5 mg I'
of inorganic nitrogen by the effluent from.the sewage treatment plant.
The flora of the stream removed nearly all of the added nutrients from
solution within the first 0.6 mile downstream from the outfall during
all periods of the year except when ice cover was present. This occurred
even during the summer months when the periphyton growth appeared to be
inhibited by unidentified agents introduced into the stream with the
effluent.‘ The organic phosphorus:phytopigment density ratio in the
periphyton was found to be more than four times greater at the station
0.3 mile downstream from the outfall than in the areas upstream from
the outfall.

During the summer months the dissolved phosphorus content of the
water decreased rapidly within the first 0.6 mile downstream from the
outfall and then increased toward the downstream stations. The area of
phosphorus regeneration was dependent upon stream flow and water
temperature.

The data indicate that periphyton production increased rapidly
after the spring thaw and reached a maximum in April or early May.

The production then decreased gradually until an extraneous nitrogen
supply produced a second maximum in June. After the June peak the
production decreased gradually until the period of ice cover. The
production levels appear to be more limited hy the inorganic nitrOgen
content of the water than by'the dissolved phosphorus content.

The standing crOp of periphyton in the stream was almost completely
dhstroyed by high water and associated high turbidities which existed

for only a short period of time.

A STUDY OF NUTRIENT ACCRUAL, UPTAKE, AND REGENERATION

AS RELATED TO PRIMARY PRODUCTION INtA WARMFWATER STREAM

by

MORRIS LEROY BRENNER

A THESIS

Submitted to the School for Advanced Graduate Studies of
Michigan State University of Agriculture and
Applied Science in partial fulfillment of

the requirements for the degree of

DOCTOR OF PHILOSOPHY

Department of Fisheries and Wildlife
1958

 

ACKNOWLEDGMENTS

The writer wishes to extend his sincere thanks to Dr. Robert C. Ball
for supervising this work. His suggestions and criticisms concerning
the field program were most helpful, and the time he spent reviewing
and editing the manuscript is genuinely appreciated.

He is also grateful for the suggestions and aid given by Drs.

P. I. Tack, E. W; Roelofs, D. W. Hayne, and.H. L. King.

The writer is indebted to his fellow graduate students, eSpecially
to Messrs. Alfred R. Grzenda, David Correll, John F. Carr, and Robin
VanNote, for their aid in the collection of data and for the many
stimulating discussions on stream biodynamics.

He wishes to thank his wife Jean for her patienée and understanding
during the data collection period and for typing the manuscript.

He is grateful for the financial assistance in the formrof a
research fellowahip extended by the Institute for Fisheries Research,

Michigan Department of Conservation.

 

iii

TABLE OF CONTENTS
PAGE
INmODUCTION . C O O O O O O O O O O C O O O O O O O O O O O O 1

METHODS AND TECHNICS

Temperature . . . . . . . . . . . . . . . . . . . . . . . 10
pHeeoeeeeeooooeeeooooooooooeee 1.0
condUCtiVityeeooeeeoeeeeeeeeoooeoee 11
T‘J-rbidityOOOOOOOOOOOOOOO000...... ll
AlkalinityOOOOOOOOOOOOOOO0.0.0.... 11
Carbon Dioxide. . . . . . . . . . . . . . . . . . . . . . ll
Dissolved Oxygen. . . . . . . . . . . . . . . . . . . . . 12
PMEmmnm... ... ... ... ... ... ... ... 12

TotaJ- Phosphoms O O O O O O O O O O O O O O O O O 12

Total Dissolved Phosphorus . . . . . . . . . . . . . l2

sestonicphosphomSoeeeeoeeeeeeeoooo 13

Acid-Soluble Sestonic Phosphorus . . . . . . . . . . 13
ArmoniaNitrOgen..................... l6
Nitrogen as Nitrite plus Nitrate. . . . . . . . . . . . . 16
Periphyton Measurements . . . . . . . . . . . . . . . 16
Spectrophotometric Analyses of the PhytOpigments. . . . . 19

DESCRIPTION OF THE AREA. . . . . . . . . .

Physiography....................... 23
Physical and Chemical Characteristics . . . . . . . . . 25
RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . 38
-NutrientMetabolism................... 39
. Phosphorus . . . . . . . . . . . . . . . . . . . . . 39
Abiotic phosphorus removal. . . . . . . . . . . hl

Biotic phosphorus removal . . . . . . . . . . . hi;
Phosphorus regeneration. . . . . . . . . . . . 146
AmmoniaNitrogen.................. 51
NitrOgen as Nitrite plus Nitrate . . . . . . . . . . 5h
PeriphytonPrOdIICtioneoeeoeeeeoooeeeoee 58
SeasonalVariationsoo............... 59
Variations between Stations. . . . . . . . . . . . . 62
NutrientResponse.................. 68
Effects of Turbidity on Stream Periphyton. . . . . . 77

SpectrOphotometric Analyses of the Phytopigments.
Projected Intercornmunity Relationships. . . . . .

SW. C C O I O O 0 O O O O O O O O O O 0

LITERATURE CITED

APPENDIX A
APPENDIX B
APPENDIX C
APPENDIX D

APPENDIX E

iv

PAGE

82
8h

89
93

LIST OF TABLES

PAGE
TABLE 1 Phosphorus values (pg I') obtained initially
and after 30 days storage with and without
aCidificatione O O O O O O O O O O O O O 0 O O O O 0 1h

TABLE 2 Location of the sampling stations used to determine
the physical and chemical characteristics of the
RedCedarRiver......o............ 30

TABLE 3 Dissolved and sestonic phosphorus values of water
samples taken August 20 and August 25, 1957 from
stations located above and below the sewage
tmatmentplantmltfan............... 142

TABLE h Hicrograms of acidrsoluble sestonic phOSphorus in
100 ml water samples from 9 stations on the Red
cedarRivereoeoeeeeeeeeeeeoeeooe 143

TABLE 5 Micrograms of organic phosphoruszrelative peri-
phyton growth ratio as detennined at 7 stations on -
theRedCedarRiveroo............... ITS

TABLE 6 Sestonic phosphorus content of the water at two
stations on the Red Cedar River as related.to water
temeratureeeeeeeeeoeeoeooeeeeee 119

LIST OF FIGURES
PAGE

FIGURE 1 Plexiglass artificial substrata and supporting
blocks used for the collection of periphyton
from the Red Cedar River . . . . . . . . . . . . . 17

FIGURE 2 Correction graph for adjusting measured phyto-
pigment absorbancy values to units related to
concentration. . . . . . . . . . . . . . . . . . . 20

FIGURE 3 Map of Red Cedar River and principal tributaries.
Overlay shows location of the 12 sampling stations
used in 1956 to determdne the physical and
chemical characteristics of the river. . . . . . . 2h

FIGURE b Red Cedar River in the vicinity of'Williamston,
IfiChiganeeoeeeoeeeeeeooeeeeeo 26

FIGURE 5 Medianrmean monthly discharge or the Red Cedar
River for l9h7-l958 and mean monthly discharge
for the August 1957 through July 1958 study
period. (Data from U. S. Geological Survey) . . . 28

FIGURE 6 Total phosphorus values for two sampling dates from
12 stations on the Red Cedar River . . . . . . . . 31
\
FIGURE 7 Total phosphorus values for two sampling dates from
12 stations on the Red Cedar River . . . . . . . . 32

FIGURE 8 Total phosphorus values for two sampling dates from
12 stations on the Red Cedar River . . . . . . . . 33

FIGURE 9 Total phosphorus values for two sampling dates from
12 stations on the Red Cedar River . . . . . . . . 3h

FIGURE 10 Total inorganic nitrOgen values for two sampling dates
from 12 stations on the Red Cedar River. . . . . . 35

FIGURE 11 Red Cedar River and the sampling stations which'
are numbered according to their distance (in miles)
from the sewage treatment plant outfall. . . . . . 37

 

FIGURE 12

FIGURE 13

FIGURE 1h

FIGURE 15

FIGURE 16

FIGURE 17
FIGURE 18

FIGURE 19

FIGURE 20

FIGURE 21 '

Dissolved phosphorus content of the water and the
phosphoruszphytopigment ratios of the collected
periphyton from stations on the Red Cedar River
for the JUly 29 to August 5, 1958 period . . . . .

‘Micrograms of dissolved phosphorus per liter of
water as determined at three stations on the Red
cedar. River. 0 O O O O O O O O O O O O O O O O O 0

Relationship between precipitation, stream flow,
and inorganic nitrOgen in the Red Cedar River. . .

.Mean daily phytOpigment production per square
dbcimeter measured at nine stations on the Red
Cedar River from August 1957 through August 1958 .

Mean phytOpigment units per square decimeter per
day for'five collection periods from stations
located above and below the sewage treatment
plantoutfalleooooeoooooeooeoooo

Dissolved and sestonic phosphorus content of the
water at eight stations on the Red Cedar River . .

Inorganic nitrogen content of the water at eight
stations on the Red Cedar River. . . . . . . . . .

Total accumulation of phytopigment units per square
decimater of substratum and the'mean available
nutrient values recorded for the period from

March 20, 1958 to August 15, 1958 at seven sampling
stations on the Red Cedar River. . . . . . . . . .

Comparison of the inorganic nitrogen content of the
‘water and.the phytOpigment production measured at
eight stations on the Red Cedar River for the
peI'iOd from April 18-25, 1958. o o o e o o o c o 0

Calculated and measured effects of high water and
turbidity on primary production and on the standing
crop of periphyton growing on plexiglass substrata
in the Red Cedar River . . . . . . . . . . . . . .

vii

PAGE

h?

50

56

60

6h

70

71

73

78

81

 

viii

PAGE

FIGURE 22 Absorbancy spectra of the phytopigments from two
stations on the Red Cedar River. . . . . . . . . . 83

INTRODUCTION

 

INTRODUCTION

One of the most difficult problems facing the aquatic ec010gist
is the finding of a method for determining and measuring quantita-
tively the physiochemical and biological processes operating in a
lotic environment. A stream is a more open ecosystem than a lake
and the factors controlling the biological activity may be more closely
related to the watershed than to the stream bed. Also, the unidirec-
tional flow continually carries the seston away from the area in which
it was produced so that the organically combined nutrients are lost
from the biotope and the develoxxuent of an autochthonic plankton
population is virtually impossible.

The wide variations in velocity, turbidity, and water level in
a stream tend to alter the substratum. The existing flora and' fauna
can be destroyed by the exclusion of light or by the abrasive action
of the bed load associated with high-water conditions. Therefore,
it is possible for the entire biological system of a stream to be
disrupted by the run-off from a single rain. One can surely agree
with Purdy (1923) who said "A large lake represents stability of
environment, but a flowing stream is the fullest expression of a
condition of instability. " '

In view of the increasing demands for aquatic recreational areas,
the need for a better understanding of stream metabolism and stress

DrOduction becomes more apparent. Paradoxically, at a time when the

 

streams are destined to carry a heavier recreational load, they are
also receiving more wastes from human activities. Nearly all strewns,
with the exception of those remote from human habitation, serve to a
greater or lesser extent as a dumping ground for biological and
industrial wastes produced by man. It is well established that streams
have a capacity for self-purification and that the physical, chemical,
and biological forces involved in the stabilization of wastes are
interrelated and mutually dependent.

Organic material added to a stream undergoes carbonization,
nitrification, dephosphorulation, etc. , and releases the basic nutrient
constituents to solution. If the amount of organic material added to
a stream exceeds its capacity for self-purification and stabilization
of the putrescible fraction, the dissolved oxygen supply in the water
is depleted. this results in the accumulation of unstabilized waste
material on the bottom of the strewn and the fomation of toxic
anaerobic decomposition products . The composition of the flora and
fauna is then limited to those species that can tolerate highly
adverse enviromnental conditions . ‘

The modern sewage treatment plant concentrates the physical,
chemical, and biological forces involved in the stabilisation of
Organic matter into a system of settling basins, aeration tanks or
filters, and digestors, and therefore prevents the formation of a
septic zone in the both of water receiving the effluent.

The nutrient content of the effluent varies with the type of \ A
treatment and efficiency of the sewage treatment plant. Also, the \
rate of enrichment of the receiving waters varies with the dilution I!

 

factor as influenced by the volume of effluent and with the precip-
itation and runroff in the watershed area of the stream. The rate
of biological response to the introduced nutrients is also dependent
upon a.myriad of adverse physical conditions which are inherent to
flowing‘waters.

The dynamics of the biological phase in enriched streams can be
studied.by measuring the production at any one or several of the
trophic levels. The presence of a delayed increase in production
may indicate that toxic or inhibitory agents are present in the effluent
which prevent the utilization of the introduced.nutrients by the
primary producers. The organisms composing any trOphic level can be
used to study the biological effects of the introduced nutrients
and associated organic and inorganic substances found in sewage
treatment plant effluent since they are either directly or indirectly
dependent upon the primary producers for food.

The difficulties involved with production measurements at a
specific level increase with the position of the organisms in the food
chain. Generally speaking, the organisms of the higher levels are
characterized by changes in food habits during their life history
whereas the members of the lower levels usually can be classified as
Primary producers or herbivoresthroughout their entire life history.

FiSh are mObile and individuals of many species tend to migrate
from.areas providing protection and cover to areas of food production
or food concentration. Also, some primary consumers such as the white

sudker (Catostomus commersonnii) which are independent of many of the

 

"Side food chains" so characteristic of the carnivores have a wine

range of movement associated with specific phases of their life
history. It is also very difficult to obtain a quantitative swnple
of a fish population. For these reasons the fish population does not
lend itself well to a study of production in streams. I

The standing crop and growth rate of the benthic fauna population
are often used in productivity studies in a lentic environment. Al-
though there may be significant differences in the species composition
and number of organisms found in different bottom types or between the
littoral or profundal zones of a lake, the number of microhabitats in
a stream bottom increases the sampling problems. On the basis of a
single uniform riffle, Neecmam and Usinger (1956) found that 191;
samples would be required to give significant figures on total wet
weights at the 95 percent confidence level and that total numbers
would require 73 samples. J

Since productivity involves the unit of time, the measurements
are complicated by variations in water level between sampling periods.
The detrimental effects of high water on the benthic fauna are well
recognized. Allen (1951) reports that a flood on the Horokiwi Stream
destroyed 85 percent of the number and 88 percent of the weight of the
benthic fauna.

The length of time period required to accm'ately detect differ-
ences in growth of certain organisms can subject the method to the
hazard of high water. Trans (personal communication) reports that 33
days were required for Stenonema gulchellum to grow one millimeter in
length when reared in the laboratory under optimum conditions.

The phytoplankton population is frequently used in limn010gi cal

and oceanographical work to study the productivity of standing waters.

 

V

Plankton can usually (be found in streams; but, as Ruttner (1952) points
out, it is impossible to distinguish between eupotam0plankton and
tychoplankton doomed to death in the lotic enviroment. Also, a sudden
rain will often flush the plankton from a strewn, or conversely, if
the strewn channel is characterized by swwnps and ozbows, may cause a
sudden increase in plankton. Butcher (1932) found that a large portion
of the plankton present in streams was detached sessile algae in the
process of decomposition. For these reasons the phytoplankton pepu-
lation of a stream is not necessarily indicative of strewn conditions.

The periphyton (-Aufzbchs) consists of the community of organisms
which grows on the stream bed and on submerged objects in the water.
Although benthic fauna are frequently found in the mat, they are not
considered as a part of this connunity.

The periphyton plays an important role in the lotic environment
because it is virtually the only primary producer in the ecosystem.
Also, because of its perpetuity and rapid turnover period, the volume
of this material produced annually in a given area is enormous. Even
thong: the organisms of this group are subjected to the some adverse
conditions as the benthic fauna or the fish, they are characterized
by a very rapid recovery. The production within this community may
also be used to study the nutrient levels of the water mass flowing
by since these organisms are not equipped with a means of procuring
the essential elements from the stream bed. Considering that the
Primary consumers in a stream are almost entirely dependent upon this
commmity, the production in the higher trophic levels can be estimated
by relating the production of the autotrophic organisms in this group
(Lindeman, 191:2).

Although Hentschel (1916) was apparently the first to employ
artificial substrate to stucb' the accumulation of sessile organisms,
the method has been virtually overlooked in this country. Butcher
(1932, 191:7) mounted glassslides in frames and submerged them in
English streams to collect sessile algae for both qualitative and
quantitative studies. Kore recently Patrick (19510 devised the well-
knoun "diatometer' for holding glass slides for the collection of
diatoms for evaluating stream conditions . A comprehensive review of
the literature concerning the use of artificial substrata for the
collection of all types of aquatic and terrestrial microorganisms is
given by Cooke (1956).

Hooper, Ball, and Hayne (ms) were the first to combine the phyto-
pigment extract method as used by Kreps and Verbinskaya (1930), Harvey
(1931.), Manning and Juday (191.1), and others with the artificial sub-
strata method for estimating periphyton production. This method has
been used and refined by several of their students in studies of
fundamental productivity in streans , the data of which are given in
the Master's theses of Grzenda (1955) and Alexander (1956).

In this method artificial substrata are exposed in a stress for a
given period of time, removed, and the phytopiment from the periphyton
M extracted with 95 percent ethanol. ‘lhe absorbency of this
solution is then determined with a photoelectric calorimeter. The
Primary production for the period can then be expressed in terms of
net phytopigment density per unit area of substratum to compare areas
Within a given strean or to compare exposure periods, or converted to
units of weight (Grzendal) by use of an experimentally determined value

‘
A

lboctorate thesis in preparation. Method reported at the meeting
or the American Institute of Biological Sciences, Bloomington,

Indiana, 1958 .

 

representing the relationship between phytOpignent density and weight
of organic material.

After a preliminary stuck of the entire water course of the Red
Cedar River to determine the physical and chaical characteristics of
the water, a 5.3 mile area located imediately downstream from the
City of Williauston was chosen for extensive study. Upstream from
this area a reservoir served as a silt basin, Just within the upper
limits a turbid tributary strewn emptied into the river, and within
the first 0.5 mile the outfall from a sewage treatment plant provided
a continuous source of nutrients without producing septic conditions.

he intensive stuw in the relatively short area was made to
determine the fate of nutrients introduced into a natural stream and
to measure the biological response as indicated by periphyton pro-
duction to different nutrient levels. During the course of the study
the seasonal changes in periphyton production as well as the effects
of adverse physical conditions such as high-water levels and associated
high turbidities were determined. It is the aim of this study to
obtain a better understanding of nutrient metabolism and the production

of organic material in a lotic environment.

METHODS AND TECHNICS

METHODS AND TECHNICS

'lhe year-around sampling program required that the water samples
for chemical analyses. be cooled in manner and prevented from freezing
in winter to preserve the chemical and biological equilibria. This
was accomplished by placing the samples in ice water in a portable ice
chest innediately after collection in polyethylene bottles. The
analyses were either completed the same day or worked to a point where
storage would not result in either a gain or loss of constituents.

All dimensional units are given according to the algebraic

exponential system. For example, 1 - a" and g/dm‘ - g dm“ .
a’
Water Temperature

Water temperatures reported for individual stations were taken
with a pocket thermometer held approximately three inches under the
water surface. The temperatures reported in Appendix A were recorded
on a Taylor recording thermometer permanently located ten miles upstream

from the mouth of the river and ten miles downstream from Williamston.

21‘.

The pH values were determined on a Beckman Model N portable pH
meter. All measurements were made in the field during the sunsner

Period or immediately after returning to the laboratory during the
"inter months.

 

Conductivity

The electrical resistance of the water was determined with an
Industrial Instrument Company Model RC-7 portable conductivity meter.
The resistance readings were corrected to 18,0 C. and converted to
ohms" cm“ x 10“ . All measurements were made at the swnpling location
during the stunner period and immediately after returning to the labo-
ratory during the winter.

Turbidity

'Ihe turbidity measurements were made imediately after returning
to the laboratory on a nett-Summerson photoelectric colorimeter which
had been calibrated with the Jackson Candle Turbidimeter. A correction
for the intrinsic color of the water was made by adjusting the instru-
ment to zero with a filtered river water sample in the light path. All
readings were taken using the blue filter having an approximate spectral
range of 1400 to h65 millimicmns.

Alkalinity

C

The alkalinity determinations were made in the laboratory using
methods described in "Standard Methods for the Examination of Water,

 

 

 

Sewage, and Industrial Wastes" (mm, mm, FSIWA, 1955).
Carbon Dioxide

The free carbon dioxide was determined from the pH and alkalinity

readings using the monograph proposed by Moore (1939) .

12

Dissolved m

The dissolved oxygen was measured by the unmodified Winkler method.
The reagents were added in the field but the final titration was car-
ried out in the laboratory. With few exceptions the‘samples were taken
between 8 a.m. and 11 a.m. Thus the values represent a period when the

levels are consistently low due to plant respiration.
mosphorus

Four physical states of phosphorus were determined during the
course of this investigation. In all instances the samples were di-
gested and the phosphorus converted to the POh form, treated with
acidified wnmonium molybdate, and the density of the blue color re-
sulting from the reduction of the phosphoruolybdate with stannous
chloride read on a nett-Slmmerson colorimeter. The method was
modified slightly from that described in Ellis, Westfall, and Ellis
09er) in that the final 100 ml solution was divided and neutralized
with saturated Nam before the final color-producing reagents were
added (Taylor, 1937).

Total Phosphorus
The total phosphorus values were obtained after the digestion and
treatment of a 100 ml sample of river water.
Total Dissolved Phosphorus

The total dissolved phosphorus was determined from a 100 ml water

8a“Pile. that had been filtered through a Hillipore Filter. The HA type

nearbrane having a pore size of 0.16 micron was used for all filtra-
tiom.

13

Sestonic Phosphorus

'me sestonic phosphorus values were obtained by difference between

the total phosphorus and the dissolved phosphorus.
Acid-Soluble Sestonic Phosphorus

The acid-soluble sestonic phosphorus values were obtained after
the digestion and treatment of a 100 ml sample of 0.01 N H2501; which
had been filtered through the pad which retained the seston from the
dissolved phosphorus determination.

Early in the program, while becoming familiar with the methods, it
was found that the values obtained from samples that had been stored
for several days were sigxificantly lower than those obtained from
duplicate samples that were analyzed imauediately after collection. An
attempt was made to determine the mechanism by which the phosphorus
was lost from solution and to establish methods of prevention of the
loss during storage.

Five liters of water were collected from the Red Cedar River at
Williamston, Michigan in a "pyrex" Florence flask. The sample was
brought into the laboratory and mixed for three hours on a “Mag-Mix“
stirrer to allow for temperature adjustment and to insure the withdrawal
of representative sub-samples. The characteristics of the water were

as follows :

pH 7.80

Methyl orange alkalinity 256 mg 1"

Carbon dioxide content 7.8 mg 1"
Conductivity 612 x 10" ohms" cm"

Turbidity 17 units.

Ten 100 ml samples were siphoned off and transferred to Erlenmeyer
flasks for immediate phosphorus determinations. Ten u oz. polyethylene
bottles were filled for storage without acid, and ten 1; oz. polyethylene
bottles were filled for storage after the addition of 0.3 ml of concen-
trated sulfuric acid. The samples were stored in the dark at room
temperature. The total phosphorus determinations were made according
to the stannous chloride-molybdate method previously described.

The results obtained from the phosphorus determinations on the
unstored water sample and those stored for 30 days with and without the
addition of the acid are given in Table 1.

TABLE 1. Phosphorus values (pg 1" ) obtained initially and
' after 30 days storage with and.without acidification.

Sample No. "0" Days 30 Days, Acid Added 30 Days, No Acid

 

 

1 55 55 us
2 Sh 62 22
3 55 56 20
h 5h 65 28
S 53 5h 39
6 58 SS 31
7 5h 58 ho
8 Sb 58 < 32
9 57 5h 33
.12 .514. .22. .22
mean Value - Sh.8 ng 1" 57.0 pg 1" 31.6 11g 1"

The results of the determinations indicate a significant loss of

Ph°8phorus from solution in the untreated samples. In order to determine

 

15

if the phosphorus remaining in the storage bottles could be returned

to solution or ”stripped" from the walls, 100 ml of distilled water

and 0.3 ml of concentrated sulfuric acid were added to each of the
“no-acid" storage bottles and allowed to stand for 21; hours. Phosphorus
determinations on the acidified solutions yielded 22, 12, 23, 18, 28,
20, 17, 16, and 18 pg 1 . One sample was lost during handling. The
mean recovery value was 19.3 118 14..

The mean recovery value, when added to the mean value for the
samples stored without acid, indicates that phosphorus escaping from
solution can be recovered with the «addition of acid.

In order to determine if the loss of phosphorus from solution
might be due to adsorption on the walls of the bottles under alkaline
conditions, two 50 ml samples of a standard phosphate solution were made
basic (pH 11.8) by the addition of one drop of NaOH and stored. Deter-
minations made 28 days later indicated that no phosphorus was lost from
solution.

At a later date the procedure was repeated and an additional set
of samples was stored after 1.0 ml of reagent grade chloroform was
added to each bottle according to methods described by Dobie and Hoyle
(1956). Total phosphorus determinations made 15 days later indicated
an average of 12 percent of the phosphorus was lost from solution during
the storage while the values obtained from those preserved with 0.3 ml
01‘ concentrated sulfuric acid agreed with the original values to within
the precision of the method.

Later, Hepher (1958) used phosphorus fortified tap water and found

no chemical changes occurred in a sealed jar on storage, but in an

l6

unsealed jar the pH increased, bicarbonate alkalinity decreased, and

the carbonate alkalinity increased as carbon dioxide was lost from
solution. He also found a loss of phosphorus from solution that roughly
corresponded to the calculated solubilities of phosphate as related to
pH and calcium ion concentration. Therefore, the 0.3 ml of concentrated
sulfuric acid {added to the sample bottles increases the solubility of
the phosphorus by lowering the pH and preventing the formation of car-

bonate ions 0

Armenia Nitrogen

The nitrogen present in the form of ammonia was determined by the
distillation method as described in "Standard Methods" (APHA, AWA,

FSIWA, 1955) .

Nitmgen as Nitrite plus Nitrate
The nitrogen in the oxidized forms was determined by the reduction
method as described in ”Standard Methods" (AH-IA, MA, FSIWA, 1955).

Periphyton Measurements

The biological sampling pregram consisted of measuring the peri-
phyton pigments that accmulated on artificial substrata suspended in
the stream over a given exposure period. The length of time the sub-
strata were exposed was dependent upon the rate of accumulation as
8°Ver'ned by the physical and chemical conditions of the strean.

The substrata employed in this investigation consisted of plexi-
81888 plates, 7 mm in thickness, having an exposed area of 1.11 dmz
when attached to a horizontal crossbar (Fig. l) . The basic adaptation

Fig. l. Plexiglass artificial substrata and supporting blocks

used for the collection of periphyton from the

Red Cedar River.

 

 

18

and method of attachment was devised with a fellow graduate student,
Hr. Alfred R. Grzenda.

The plenglass substrata were collected after the periphyton growth
was plainly visible but before a dense mat had formed that would be
subject to slougring due to a layer of dead cells adjacent to the plastic
or to the formation of gas bubbles under the mat. The substrata were
removed from the stream, placed in individual plastic bags, and frozen
to aid in the release of the biological growth from the plastic and to
rupture the plant cells to facilitate the phytOpigment extraction. The
periphyton growth was scraped from the substrate and allowed to stand
in 95 percent ethanol for a minimum of 118 hours while stored in total
darkness. Tests indicate that samples can be stored in this manner for
as long as 30 days without a loss of phytopigments due to decomposition.
The samples were filtered through glass wool and the volume of filtrate
adjusted to 50 ml by either dilution or evaporation. The color density
of the ethanol-soluble phytopigment solution was read on a nett-Summerson
calorimeter using the red filter (610-700 ml).

Experiments dealing with the opticochemical characteristics of 95
percent ethanol phytopigment extracts show the absorbency of broad
Spectrum light (6h0—700 rap) is not lineally related to the concentration
of the pigments except at very low values. The deviation becomes apparent

at approximately 100 units when read on a Klett-Summerson calorimeter
and increases prOportionately with higher concentrations. This deviation
from the Lambert-Beer Law may be due to an interaction between the
solvent and the solute or to changes within or among the molecules.
This flattening of the curve when absorbency is plotted against phyto-

Pigment concentration destroys the correlation between measured pigment

l9

density and the weight of the organic material from which the pigments
were extracted (Grsenda, unpublished).

The measured pigment density may be corrected to correspond with
the theoretical absorbancy as related to concentration, and the corre-
lation with the weight of organic material restored by determining the
deviation from the Lambert-Beer Law. A correction graph was constructed
by plotting absorbancy versus concentration as determined by dilution
and by concentration of a phytOpigment solution (Fig. 2). In making
the correction the measured density is found on the ordinate, followed
across to the intercept with the experimentally determined line, and
read vertically to the intercept with the extrapolated straight line.
The absorbancy unit Opposite this intercept represents the corrected
reading.

In order to avoid confusion between the measured and the corrected
absorbancy values, the unit of adjusted absorbancy, designated as (M),
was adopted for the latter values. The values were then multiplied by
10" to avoid the use of the decimal point. Therefore, the corrected
absorbancy units of the extracted phytopigments are given as AA 1: 10' .
Since these units are lineally related to the weight of periphyton
PI'Oduced, the term phytopignent unit (optical density or absorbancy of
One AA x 10’) is used as the index of organic material production.

Spectrpphotometric Analyses 93: the Mopignents

Periodically, qualitative spectmphotometric analyses were made
°f the phytopigment extracts from stations located above and below the

sewage treatment outfall. The pigments from a portion of the periphyton

Fig. 2. Correction graph for adjusting measured.phytopigment

absorbancy values to units related to concentration.

 

 

ABSORBANCY

5.0

'o

u
T1111:

 

 

 
   

lllllJL l llllll

 

 

I0
RELATIVE CONCENTRATION

I00

21

from a substratum were extracted with 90 percent redistilled acetone
(Richards with Thompson, 1952) and the absorbancy spectrum plotted
from 1400 to 700 millimicrons.

DESCRIPTION OF THE AREA

DESCRIPTION OF THE AREA

2112225192121

The Red Cedar River, a warm-water stream, is a tributary of the
Grand River system and drains the south-central portion of the Lower
Peninsula of Michigan. The stream originates as the outflow from
Cedar Lake located in Marion Township, Livingston County in Sections 28
and 29, Township 1 North, Range 3 East of the Michigan Meridian and
flows through or near the communities of Fowlerville, Webberville,
Williamston, and Olcemos before entering the East Lansing and Lansing
areas and the confluence with the Grand River (Fig. 3). The total
length of the main channel is approximately 1:9 miles and the drainage
area is approximately 1:75 square miles. The river has a mean gradient
of 2.5 feet per mile with one-half of the fall occurring in the upper
third of the channel.
There are three dams which form artificial impoundments on the
Red Cedar River. The largest, located within the City Limits of Wil-
liamston, was originally constructed in 18h0 to provide. water power
for a smill. The present structure creates a 13 foot workingbhead
01' water to Operate a constant-flow generator to provide electrical
Pointer for a private refrigeration and frozen food plant. The pool
created by the Willimton Dam is approximately two miles in length,
but for the most part is contained within a narrow belt along the main
channel.

335”.» o5 Ho congressman". Hoods—ego
one Hooaassa one oceanosoe op emaa ca cons nsosuoao assesses «H one do

coaosooa arose asshoso .aoshaesnaha ”resonate one hoses assoc can no as: .m .maa

 

.0.

can pp 3 v

34.2. seen.
a e i

. N

x

\>

\I.

~

oat-......» Sluice v...

I
wIIIVTV)

 

0

u; ...) woo u!
.

”a

1‘3”)

”'0.

"0

s’.‘ 50.00 ‘03 la Gain.

.'0

...-....

[Orgd

m H:
n

H».

iv 0
O

. ....
HHH 3136' a e ..

......1!
e H

:1; :14

V

0232‘;

...
.‘

 

 

H
H
2 a «H /
/ /E
N NH/ HHH4/ HH</ /
c /
HQ
.ao>flh cap no mowpmwhopomhmno HmoflEozo

can wanna on» mewsaopoo op omma cw eons mdoaumvn wadHQEMm NH one Mo

coMpJUOH mzonm zwaam>0 .mefihwpsnwnp HdmfioGALQ cam po>fim umeoo com me not .m .mwh

ith is. '0. so... 3.00 Cells;

 

 

 

Ou‘
o
I
.
0 O
1 . I
s v N z
0 1 V
.I V I
0 I
. I Q Q
no ~ . I S.
1. n a. a a
I
. ‘ I. . ‘
I 1
I .0 C I
0
0 I 0 1 . 0
. . U ' I
I
A...) 4
u a so... 34; menu; a i a
. I
’3" a . a
. Ir. .
I s C E. I
89;.ng 0 it
_ I a o h m
. 0 O
1 n 4 a a
v . “8'3 0 H
I, .
O .
0838(4 u \
.U H T Q. Q
. ? ...
ga HICJ 02_fl2(4

 

 

25

The original dam in Okemos was constructed to provide water power
to operate a gristmill. The original structure has been replaced by a
small stone ballast type that creates a small pool for recreational and
aesthetic purposes. The East Lansing dan, located within the campus of
Michigan State University, is constructed of concrete and maintains a
constant depth of water for recreational purposes and for a cooling
water supply for the steam-generated power plants on campus.

he bottom material of the Red Cedar River consists of fine sand
in the upper regions to rocks and gravel with small areas containing
silt deposition in the middle and lower stretches. In general, the
bottom gradient is uniform with long pools or runs rarely exceeding
four feet in depth divided by short riffle areas. The shoreline slaps
is gradual and covered with vegetation, resulting in very little erosion
or cutting above the normal water line (Fig. )4).

he topOgraphy of the watershed area is nearly level to rolling
as a result of the Cary phase of the Wisconsin glacier. The soils of
the area are classified by Whiteside 9.1.5. 51. (1956) as being derived
from liny loam glacial till. The primary soil series are of the Miami
and Conover types having good to intermediate drainage. A large pro-
portion of the watershed area is used for dairy cattle grazing and small

grain farming.
Physical and Chemical Characteristics

Two permanent stations are maintained on the Red Cedar River to
record the physical characteristics of the stream. The first, a gauging

station five miles upstream from the mouth, is maintained by the United

Fig. be

Red Cedar River in the vicinity of Nilliamston, Michigan.

 

____1
M _- bad—s..—

 

 

 

tr: e

 

27

States Geological Survey to record the rum-off for the 355 square mile
area exclusive of the Sycamore Creek Drainage.

The flow is usually the greatest during March or April men the
combination of melting snow, frozen ground, and spring rains occasion-
ally result, in floods that cause considerable property damage. The
maximnn discharge recorded at this station during the 1931-1958 period
occurred in April 19117 when a flow of 5510 cubic feet per second was
recorded. The minimum recorded during the above period was 3 cubic
feet per second on July 31, 1931.

'me median-mean monthly discharge for the 19147-1957 period and
the rather atypical data for August 1957 through July 1958 are given
in Figure 5.

The graduate students in Limnology, Department of Fisheries and
Wildlife at Michigan State University maintain a temperature recording
station at Dobie Road, 10 miles upstream from the mouth of the river.
The daily high and low water temperatures for the period from July 1,
1957 to July 31, 1958 are given in Appendix A.

Twelve sampling stations were chosen along the main channel of the
Red Cedar River within the area from h.5 to 311.7 miles from the mouth
to stuch' the general physical and chemical characteristics of the water.
Twelve series of samples were taken during June, July, August, and
58ptember 1956 and the following data were collected:

1. Air temperature 7. Dissolved oxygen

2. Water temperature 8. Total phosphorus

3. Specific conductivity 9. Nitrogen as ammonia

h. Alkalinity 10. NitrOgen as nitrite plus nitrate

5. pH a 11. Gauge height at Mile 5

6- Turbidity 12. Discharge at Mile 5.

Reina Hmowmoaooc
6522.“ See sonata seen 33 .22. sweeten $3 enemas 2: .8.“ emfifiefl
at Ahflnenos_auos and mmmanwzmfi non am>wm Havoc pom one no omawnomap hanwsos quoeusswnoz .m .mwm
3x. 95—. an: :34 .3 .nem .5... 68 .5. $8 .38 .mnu

‘ 1‘ '

 

‘ ‘

 

 

1emhunonwd . .
Same use: 3.32.. .. to...

usage: 3:33 a ...II .

 

§. § §

puooeg Jed qseg 0]:an tr; aflnqasm

§

 

 

 

29

The values for this series of samples are given in Appendix B.

The specific locations of the twelve stations are given in
Table 2.

It will be noted from the overlay for Figure 1 that the station
sites were selected to make possible the measuring of nutrients from
either tributary streams or municipal sewer outfalls.

The waters of the Red Cedar River are highly buffered and show
little variation in pH. The bicarbonate ion content (expressed as
CaCOB) ranges from 250 to 300 mg l" and fluctuates inversely with
stream flow. hep}! values ranged from 7.1; to 8.3 during the initial
survey period.

The high bicarbonate ion content contributes to the high specific
conductivity of the stream. Values ranging from 372 to 590 x 10" ohms"
cm" were measured during the 1956 period with the wide range due to
variations in water levels. An increase in the specific conductivity
was noted toward the downstream areas. In view of the uniformity of
the methyl orange alkalinity values between stations, the increase in
specific conductivity as the water mass moves downstream can be at-
tributed to the addition of other soluble salts.

his greatest change in chemical characteristics was detected in
the basic nutrient elements, phosphorus and nitrogen (Figs. 6—10).

The data indicate that municipal effluents are responsible for the
greater portion of the nutrients introduced into the stream and that
the tributary streams contribute very little to the nutrient budget.
Sinbe Lund (1950) found that the aquatic flora can utilize phosphorus

in concentrations as low as one microgram per liter, this element might

30

095.5 poem oHHEeHsom 50.5 5293: Tam HHM
ewefim seem nenem 5» and 3333: «in HM
ownaum neon mnemoao seam escaped: m.Hm N

empflm seem 353 son.“ sensed: 13 an

tweets S 3&3 sameness seem antenna mam n:
5a savanna; and 2333.8 eds m6 92” Hg
8353.3: 5 spoke sentence: to 63 E
omens seem neg Eon.“ sensed: 93 >

omens 3 353m .m .a spa steeped: 0.2 E
omens teem 398 seen steeped: _ QB HHH

8286 5 sepia SHE 5%.: not.“ sensed: m.» HH
. pea minted 533m

3.3250 passe 5320535 33m sewage; one. so @333 m4. H

 

 

 

5.333 5502 saw neg nofispm
. £an .808 com on» no
333933.35 defiance p5 deowwba 23 05.53% on pens e533» wag—an «o dogwood .m an.

335m 58 cw: 23 co mcoflmpm ma SOC 89% mcwamfimm 25 you 025d» ashosnmona H.309 .0 .mfim

 

 

 

 

1
3 nova o 5.5: 89G 3.3:
mm on 3 cm ma 2 m o
/
v / /o/ . oUd‘hUl‘loljoA . CM
’0 \ 0 [III \\\\
. ///l \\\o// I’ll‘sxs L 00
w ///o\\\\\ A om
.m
. ... ! ONH
M
. .. . omfi
m m
n J
v . n A om.” I
n -
n
V n . OHN
.
W
. <. 35
v 4 05m
. . 8m
33 .mw 82. .. ---
omfi .3 82. n II . omm

 

 

 

 

 

32

 

 

 

 

 

835m .3an nmm 93 so mcofimpm NH 20.5 mafia mfiaafim $5 How macaw» mgofimozm H309 J. .mfim
umbwm Ho 5.32 sou.“ 3.3:
mm on mm on ma 3 m o
v L
a A
v .
v A
v A
v A
. k
32 .2 22. . --....
. om? .m 32. .. I... .

 

 

on

00

02
03 m
8H 1.
3m
3m
08

com

0mm

3'3

383m Havoc Umx 9.3 no msodtwpm NH 50.5 89% mafiamsmm 25 pom mmsdmh mgofimofi Haven. .0 .mfih
89,2 Ho 5902 Son.“ mag
mm on mm ow ma 3 m o

 

 

owH

03

L

OCH .1.

A

0.3

, 04m

 

4 2w

QmQH «hm 09,.qu I --....a

..T $2 .5 £35 - I. A 02

 

 

 

3h

..Hmbwm .2250 cam 23 no mcofimpm NH £95 3me manage» 0.5 you $3”: agonanofi H309 .m .35“

mm on

35.x Ho 58: £95 mag
mm Cu m.” 0H m o

 

 

E .om Hanamfiom .. 1--
3 6m noneSQom - 1|

 

. ' ‘

“““I‘

O\. 01
o \\ \ I
\\
\\
/ \\

 

 

A

A

A

 

 

 

 

 

on

00

RH
oma m
8H J
05
SN
03

com

0mm

 

 

.ho>wx pmnmo vmm on» :0 macapmpm

«H sogm mmpmv mcwaaswm 039.50% mmzfiw> cowoupwc

 

 

aficawpocfl Hmpoa .oa .mfim
a.» hmbfim mo 5:0: Son.“ 3::
mm on ma ON ma 0H m o
. L
. //// ./\\Jl\r\.uu\!.uV1|luuuuuu.ulnaucu|I .\\I / . .
{I \\s\ E.
hm II.\ o. \\\\ \\\\\\\\ 0" 'I01
w. \
.. x .
f m . .u
. .
u .
.
v . moo

 

mad .om nonampnom u -za.
omma .NH ouamq< . In:

 

"-~---------—---- --¢—-— -..---- ----

-".
---.'
--"_—'- rC—-—’---’-
—’---
'—'

otn

 

 

”I x 3m

always be considered as being present in the Red Cedar River at above
the minimal level. The available nitrogen concentration was found to
be relatively low in certain areas and may be a limiting nutrient for
primary production in the Red Cedar River.

The greatest accrual of nutrients occurs within the Nilliamston
City limits. During the initial survey period in the summer of 1956,
before the sewage treatment plant was put into operation, nine tiles
with an estimated total discharge of 100 gallons per minute fed waste
water, raw sewage, and septic tank effluents into the stream. As the
interceptor system was completed, the flow was diverted from these tiles
to the sewage treatment plant. me effect of the diversion was apparent
in the total phosphorus values a. plotted in Figures 6 through 9.

In order to stuck the bio- and abiodynamics of the stream in the
vicinity of the Willianston sewage treatment plant outfall, seven
permanent and two temporary stations were established within the area
from one-half mile above to 11.9 miles below the outfall. These stations,
described by the distance in miles above or below the outfall‘, were
designated as -O.h, -0.2, +0.1, +0.3, +0.6, +1.3, +1.8, +3.7, and +b.9
(Fig. 11). During the period from July 1, 1957 to August 15, 1958, the
study was confined to this 5 .3 mile area above and below the source of
nutrients. Two stations were maintained upstream from the outfall.

This was necessary to determine the chemical and biological effects of
the Deer Creek drainage on the Red Cedar River. I Thirty sets of water
sanples were taken for chemical analyses during the above period to study
the. relative enrichment, uptake, and regeneration of the nutrient elements
and the periphyton production was measured for 2h periods by the phyto—

pigment method previously described.

flue-1

37

cousamwp Manna op mcwphoooa coaches: was news: msoapwpo msaaaesm esp was ao>fim sauce com .HH .wwm .

.Hmevso pssaa unusuaoAp amazon ans Bonn Aucaas :wv

 

 

m.H+

 

 

odd a . 38m

 

if
\\ /5\\\ H X/

(I,
I

Juneau 58 com i,

 

 

 

 

 

RESULTS AND DISCUSSION

 

RESULTS AND DISCUSSION
Nutrient Metabolism

The mode of fertilization in this study differs from the artificial
enrichment of a stream as reported by Huntsman (191:8) in that the
nutrient supply is in the form of sewage treatment plant effluent and
is contirmously supplied to the stream. This may result in the flora
immediately downstream from the outfall becoming physiologically adjust-
ed to high nutrient levels. The absence of toxic industrial wastes in
the effluent and the inexistence of a septic zone in the stream should
prevent the modification of the biocoenose and limitation of species as
reported by Butcher (19h?) or Blum (1957). Theoretically, the flora
in the stream immediately downstream from the outfall would remain bio-
logically "healthy" and capable of carrying on the normal metabolic
functions. A

Phosphorus

The study of the physical and chemical characteristics of the Red
Cedar River indicated that municipal drains and sewage treatment plant
outfalls are the most important sources of phosphorus. Although the
tributary stream introduced water of high phosphorus content during
periods of high flow, the adverse physical conditions which accompany

the influx of nutrients prevent utilization and biological response

by the flora of the stream.

to

The Williamston sewage treatment plant has a mean base effluent
output of approximately 150,000 gallons per day. Since the interceptor
system is a combination storm and sanitary type, the output and dilution
vary with the local precipitation and runroff. Total phosphorus deter-
minations were made on effluent samples collected by plant personnel on
October 7 and.10, 1957 to determine the total phosphorus content of the
effluent after it had passed through the primary treatment plant. No
precipitation occurred during this period and the effluent discharge
was not diluted.by storm.water. The results indicate a high variation
in phosphorus content depending upon the time of the day and the day
of the week. The 8 a.m. samples contained 7.1 and 7.8 mg P I”
respectively on the two dates. The highest value (23 mg P I” ) was
found in the sample taken at 2 p.m. on.Monday, October 7. The frequency
of occurrence of each phosphorus level in the effluent was estimated
for the entire week. The daily, weekly, and annual phosphorus dis-
charge with the effluent into the Red Cedar River was then calculated
from.the mean phosphorus value.

The mean total phosphorus output per day was computed to be approx-
imately 6,800,000 milligrams. This is equivalent to 2.h8 metric tons
per year or an annual per capita phosphorus_output of 8.5 x 104metric
tons. Although Rudolfs (19h?) gives the phosphorus output per capita
in sewage effluent after primary treatment as 2.21 x 10"metric tons
per year, the calculated value for‘Williamston does not seem unreason-
able since the sewage is more concentrated due to the absence of heavy
industrial water users in the city and the increased use of household

detergents which are high in phosphorus.

The level of phosphorus acquired by the Red Cedar River as the
result of receiving the sewage treatment plant effluent varies with
stream.flow; but when the flow at'Williamston is less than 50 cubic
feet per second, it exceeds 100 pg 1" .. Approximately two-thirds of
the total phosphorus discharged into the river is in the dissolved
form, but it may be combined in many complex inorganic and organic
compounds. Although many complex forms of phosphorus are not immedi-
ately available to the aquatic flora, they may undergo hydrolysis or
dephosphorulation as the result of bacterial action and.be converted
to an available form (Harvey, l9h0).

The results of the chemical analyses indicate that nearly all of
the dissolved phosphorus introduced by the effluent of the sewage
treatment plant are removed from solution within the first 0.6 mile
below the outfall. In order to determine the rate of removal, midstream
water samples were taken during a low water period on August 25, 1957
at locations 50, 100, 200, 300, um, 500, 600, and 700 yards downstream
from the outfall and the total dissolved.and total sestonic phosphorus
content detenmined. The results, along with those obtained August 20,
1957 for upstream stations -0.2 and -0.h, are given in Table 3.

Even though the effluent is not completely mixed with the river
water after flowing 50 yards downstream from the outfall and.after pas-
sing through a abort riffle area, the reduction in the dissolved phos-
phorus far exceeds the theoretical reduction calculated for dilution.
This indicates that phosphorus is being removed from solution either
'ny chemical precipitation or biological uptake.

Abiotic phosphorus removal. In a highly buffered stream having a high

pH one might theorize that the soluble phosphorus combines with the

TABLE 3 .

 

 

Dissolved and sestonic phosphorus values of water samples
taken August 20 and August 25, 1957 from stations located
above and below the sewage treatment plant outfall.

Sampling location
0.1; mile upstream1

0.2 mile upstreaml

50 yards
100 yards
200 yards
300 yards
1400 yards
500 yards
600 yards

700 yards

Samples taken August 20, 1957.

downstream
downstream
downstream
downstream
downstream
downstream
downstream

downstream

Dissolved phosphorus

51 HST

h?
130
1:5
145
SO
39
50
St
146

Sestonic phosphorus

. 72 n8 1"
149
101

63

62

73

66

6h

73

61

calcium to form one of the insoluble complexes and be precipitated

from solution.

The relationship between phosphorus loss and alkalinity

of the water in a lentic environment has been described by Barrett (1952).

In a stream characterized by continual agitation the inorganic '

crystals of a calcium-phosphate complex would remain in suspension

for some distance downstream from the point of formation. Upon filtra-

tion of a water sanple, these crystals would be retained on a Millipore

filter pad and, after being removed by a dilute acid solution, could be

measured quantitatively by the method previously described.

The acid-soluble sestonic phosphorus was determined for seven

periods during the period from July 1, 1957 to July 31, 1958. The

results are given in Table 1;.

 

h3

 

 

m.H o.H
o.m 4.H
m.a ~.H
N.H N.H
m.a m.H
~.H ~.H
m.4+ m.n+

m.H
ed
m.~
m.m
«d
«d

~.H

 

m.H+

:.m.

~.m
0.H
CH+

o.m
m.H
m.m

N.N

H.m

o.a

 

0.0+

 

833m

o.~
e4
m.m
at,

:.m
o.~

 

m.o+

w.m
:.m
o.~
a.m

a.»
~.m

 

H.o+

m.H
m.H
m.m
m.H
H.m

H.m an

;.m mm

ed H has
H.m , ma

on m seas
o.m em

m.4 mmma .ma an:
mam” mam

.aopam edema com on» so
neowpmpm one: scam endgame hope: as 00H ma asaonnnonu escapees camsaomnpaom mo msmawoaoaz_ .4 mgm<a

.| lull-itlllltllu i.I «VI.

 

 

 

The results of the analyses indicate an increase in the acid-
soluble fraction at Station +0.1, but the values at Station +0.3 were
comparable to those found for the stations located upstream from the
outfall. The data would appear to indicate that although a small amount
of acid-soluble inorganic phosphorus is in suspension immediately below
the outfall the reduction in soluble phosphorus cannot be attributed to
precipitation.

A large amount of euplankton flows over the dam from the reservoir
upstrean from Station -0.h during the summer months and the amount of
plant material in the water as indicated by the green color of the fil-
ter pads shows a gradient from the upstream to the downstream stations.
With the exception of the slight increase at Station +0.1, the reduction
of acid-soluble phosphorus appears to be closely related to the decrease
in plankton in the water. Apparently a portion of the phosphorus in
the decomposing limmplankton is acid-labile after the cells fail to
survivethe rigors of the lotic environment.

W phosgzorus removal. Having failed to demonstrate an abiotic
mechanism responsible for the reduction in phosphorus in the area im-
mediately downstream from the outfall, the flora of the stream was in-
vestigated.

Although the rooted aquatic plants and trees along the shore un-
doubtedly obtain a portion of their nutrients from the river, the peri-
phyton of the stream is in contact with a greater part of the water mass
and would be more likely to be responsible for phosphorus removal.

The quantitative periphyton study described later indicated that

even when the increase in periphyton production occurred in the areas

b5

several miles from the outfall, the decrease in dissolved phosphorus
between Stations +0.1 and +0.6 remained relatively constant. If the
periphyton in the enriched area was storing phosphorus to levels above
its normal requirements as found by Einsele (19h1), the mechanism for
the removal of the element from solution even during periods when the
periphyton production was low could be detected.by determining the
phosphoruszweight ratio of the growth on the substrata from.the different
stations.

In order to determine the phosphorusrweight ratio of the peri-
phyton, total phosphorus determinations were made on the growth on one
substratum from eadh set at each station except +0.1. The latter was
eliminated from this portion of the study because the results might be
erroneous due to nutrient-rich particulate material filtered from the
water by the periphyton growth. The periphyton on the remainder of the
substrate from.each station was treated.to determine the relative amount
of growth. The results are given in Table 5.

TABLE 5. Micrograms of organic phosphorus:re1ative periphyton

growth ratio as determined at seven stations on
the Red Cedar River.

+h.9

§222£22. P h i ent unit
-0.h 0.72
-o.2 0.5?
+0.3 2.50
+0.6 1.62
+1.8 0.9?
+3.8 , 0.75

0.6h

h6

The ratio of phosphorus to organic material in periphyton increases
greatly after the introduction of the nutrient-rich effluent and then
decreases toward Station +h.9. It is interesting to note that the de-
crease in the phosphoruszorganic material ratio between Stations -O.h and
-0.2 follows a decrease in the dissolved phosphorus in the water (Fig. 12),
but after 0.6 mile below the outfall the dissolved.phosphorus in the
.water increases but the phosphoruszorganic matter ratio continues to de-
crease. Although the data for the stations downstream from Station +0.6
may appear anomalous due to the increase in dissolved phosphorus and de-
crease in the phosphorus content of the periphyton, it should be pointed
out that the values derived from a chemical analyses of the water are a
measurement of the elements not utilized by the flora of the stream. Also,
a dynamic population of periphyton does not tend to accumulate phosphorus
above the needs of the organisms, but instead utilizes the element in
the production of new cells (Grzenda, unpublished). It may be assumed
then that the greater portion of the phosphorus introduced with the ef-
fluent into the Red Cedar River is biologically removed from solution
within a very short distance from the sewage treatment plant outfall.
Phosphorus regeneration. The increase in dissolved phosphorus down-
stream from Station+0.6 (Fig. 12) raises a question concerning the
mechanism of phosphorus transport from the area of "high" concentration
through a distinct area of "low" concentration where the dissolved phos-
phorus values may at times be nearly equal to those upstream from the
effluent discharge to the downstream areas where the values increase

with distance from the outfall. One might assume that the downstream

b7

.eoaaoa

32 .m unease 3 am one. one see scram emcee com one. he shouted some consented ooeoefloo

oping quamfitdoifiugzsndoquoqg

....

no heaven pcmswaaophmaunshonanoea one one hope: on» no essence manomAnona popaouman
Haeupso scam scam:

m a m

N

bl

0H

m

.NH .wwh

 

f

 

‘ 1‘

owaaa
pacemaaophsanosmoennomm t .....

32 Am 3:. .. ..II
nanoseconm cohaommen

 

 

 

 

 

OH

ON

on

o:

om

00

0»

0w

om

OOH

sumoquoqg pantosqu .1 8d

LB

increase was due to the mineralization of particulate organic waste
released in the effluent and that the area of stabilization would move
upstream as the water temperature increased and downstream as the water
temperature decreased. Although the zone of phosphorus regeneration
tends to shift with water temperature, the method of transport cannot

be associated with suspended solids. The total sestonic phosphorus
values for the station 0.3 mile below the outfall were usually comparable
to those upstream from the outfall. Therefore, it must be assumed that
the phosphorus is transported biologically from the areas of "high"
concentration to the zone of regeneration.

It was demonstrated that the periphyton 0.3 mile downstream from
the source of nutrients contained over four times as much organic phos-
phorus per unit of growth as the periphyton.upstream.from the outfall.
One can also assume that this ratio increases between Station +0.3 and
the outfall. The quantitative periphyton data indicate that the net
amount of organic material produced may exceed 20 mg dm“ day" in
the more productive areas of the Red Cedar River (Grzenda, unpublished).
If it were not for a relatively short turnover period, i.e. sloughing
and repOpulation of an area, the stream bed would soon be covered with
a thick mat of periphyton. The sloughing process results when new
growth excludes the light from.the basal cells causing them to die and
‘become detached from.the substratum. The disintegration of the basal
cells may be accelerated by the nascent oxygen produced by the outer
photosynthetic layer whose growth in turn may be stimulated by the
carbon dioxide and nutrients released from the basal layer.

The downstream increase in dissolved phosphorus results when the

periphyton containing the high phosphorus :weight ratio becomes detached

149

from the substratum, is carried downstream by the current, disintegrates
and undergoes decomposition, and releases the nutrients to solution.

The area of nutrient regeneration is governed by the stream velocity
and the water temperature.

The role of water temperature in the decomposition rate of the de-
tached.periphyton is well demonstrated in Table 6. On August 19, 1957
and June 19, 1958 when the daily high for the water temperature approached
20° 0., the sestonic phosphorus content of the water was much lower
at Station +3.7 than at Station +0.6. On these dates the regeneration
was most apparent as shown in Figure 13. On January 1h, 1958 and
April 15, 1958 when the water temperature was below the Optimum for
bacterial activity, the sestonic phosphorus values were equal for the
two stations.

TABLE 6. Sestonic phosphorus content of the water at two stations
on the Red Cedar River. as related to water temperature.

 

 

23355. Hater tempe raturg Station
E...“ M :93} 3.3.21
August 19, 1957 17.9° c . 20.60 c 59 11g 1" 26 11g 1"
January 11;, 1958 0.0° C 0.0° C 26 pg 1" 26 pg 1‘
April 15, 1958 - 8.80 c 13.u° c 33 n8 1" 33 pg 1"
June 19, 1958 16.80 c 19.h° c 52 pg 1" 28 pg 1"

During the winter months when bacterial activity is greatly reduced,
the area of greatest phosphorus regeneration was downstream from the
study area. Mr. Alfred R. Grzenda, a fellow graduate student who was
studying the biodynamics of the stream at an area 9.9 miles downstream

from the outfall, consistently obtained higher values than were obtained

SO

.eeeem tween eem esp

 

 

so mcoapoom conga pm nonfismopow mm swam: we hoped hog msaosmmosa pcpaommwv mo nsmamoaoaz .MH .mfim
sync:
wmma pmma
.w:< hand muse no: .ua4 .amz .nom .nww .ooa .>oz .poo .pmom .m54 haze
: . OH
: ._ ON
./ .
A. o /o\0 . om
. l .
. . 01...: . . OJ 3
. h T»
u. o... m no I X/ o .... ._ 0m a
A “W; ... x . ..‘e ... fl.
. M ax ’/ ... n W
m. l x m co "a
m .7... o. .. m.
. K. I ”a Max... M 1 ON.
x xx W W“
. ... /, ... . Ow 7%
,, ... m. w.
l 5’. e— xcl; ..u . s cm W
x m
. / ... . 8H
’/ W.
xxx. . cad
e
. I :o a |.II
. m o «9 pm . ONH
0.0+ coflampm I ......
l e.m. eoaeeem -.:-- . oma

 

 

 

51

at Station +h.9. After the end of the period of ice cover (March 3)
the periphyton growth was stimulated by the additional light reaching
the substratum and the dissolved phosphorus content of the water was
reduced in the downstream areas. After the water temperature had
increased sufficiently to stimulate bacterial activity, the zone of
regeneration again shifted upstream to within the 5' .3 mile study area
and the. phosphorus accrual exceeded biOIOgical utilization. This re-
sulted in an accumulation of the element in the dissolved form and pro-
duced the downstream regeneration curve as shown in Figure 12.

Ammonia Nitrogen

Although ammonia nitrOgen is seldom a constituent of streams, it
was of great importance in this study because ammonia is the first
inorganic nitrogen form produced in the nitrification of organic mate-
rial and is usually present in sewage treatment plant effluent. Also,
the reservoir located above Station -0.h contained large amounts of
plant material and organic depositions which proved to be a source of
amonia for the section of stream under investigation.

In streams that do not receive organic wastes the ammonia produced
is almost imnediately utilized by the aquatic flora or converted to
nitrites and nitrates. The latter is accomplished through bacterial
action. The ammonia present in water is. mainly in the form of NH;
and undissociated NaOH. At the normal summer pH values of the Red
Cedar River the ratio between Mi; and New would be approximately 30:1.

the amount of ammonia produced in the Williamston sewage treatment
plant and emptied into the Red Cedar River with the effluent is high
for this type of installation. This might be explained in part by

the plant's operation at only 60 percent of designed capacity resulting
in a longer retention period of the liquid fraction in the primary set-
tling tank. Also, the sludge must be allowed to accunmlate for a longer
period of time to obtain an adequate amount to justify withdrawal and
transfer to the composting unit. The ammonia nitrogen values obtained
at Station +0.1 ranged from trace amounts during the July 1957 flood
period to 1.2 mg 1" during the low water period in July of 1958. The
effluent is not completely mixed with the river water at Station +0.1,
and the ammonia nitrogen content of the water is reduced by dilution,
conversion to nitrites and nitrates, and plant utilization within the
next 0.2 mile so that the highest value received at Station +0.3 was
0.1.;2 mg 1" . -

During the summer months when bacterial and periphyton activities
were high, significant ammonia nitrogen values were recorded only at
Stations +0.1, +0.3, and occasionally at +0.6. One exception to the
above was noted during the study period and this was attributed to an
industrial accident approximately 15 miles upstream from the area.

On May 21, 1958, a fire destroyed a portion of a metal plating
factory located at Fowlerville, Michigan. In the course of fighting
the fire, water flooded the cyanide tanks and this chemical was diverted
into a waste treatment lagoon. The plant manager quickly ordered the
lagoon outlet closed and initiated treatment of the lagoon contents
with sodium hypochlorite to oxidize the cyanide to the relatively non-
toxic cyanate (Eldridge, 1933; Dobson, 19M) and thus prevented what
might have been a major catastrophe to the stream. When the reaction

was complete, the contents of the lagoon were fed into the Red Cedar

53

River. Since cyanates are hydrolyzed to ammonia compounds, the rate

of which is a function of the pH of the receiving water, it is theorized
that this reaction accounted for the presence of ammonia at both the
upstream and downstream stations within the study area during June 1958.
Although Resnick gt;§l. (1958) found that the cyanates were relatively
stable under aerobic conditions, it is theorized that the formation of
ammonia may result from.microbial action in the complex natural environ-
ment.

During the fall and winter months when the biolOgical activity in
the area was low, ammonia nitrOgen was found at all stations. The
periphyton data indicate that when the phytopigment production fell be-
low approximately five units per square decimater per day the ammonia
production exceeded utilization and conversion.

The nitrogen introduced into the Red Cedar River in the form of
ammonia cannot be followed through the uptake and regeneration phases
that were associated with the phosphorus. Even though the quantity
of ammonia received by the stream is high during the summer period, it
is unlikely that it would be lost to the atmosphere in the form of gas
since 800 volumes can be absorbed in one volume of water at 20° 0. Also,
since the amount of nitrogen in the oxidized foms does not increase at
Station +0.6, it must be assumed that during the stunner period the
ammonia nitrogen is assimilated into organic material within.a short dis-
tance downstream from the outfall.

'nle nitrogen cycle in the study area will be covered more fully

in the section discussing nitrates.

 

I‘ll, ll! KNIIII W

.111 I

5h

Nitrogen as Nitrite plus Nitrate

The nitrate ion, unlike the phosphate ion, is not firmly held by
soil particles and is easily leached from the topsoil and eventually
finds its way into lakes and streams. Schmidt (1956) in his work in
Minnesota reports finding 6.31 mg l" of nitrates in spring water from
Nobles County and 18 mg II in water from a field tile draining culti-
vated land. In view of the above data one might expect a stream re-
ceiving not only drainage water from agricultural areas, but also
sewage treatment plant effluent, to contain large amounts of nitrates.
However, as Whipple (19148) points out, the nitrogen measured in a stream
represents only that which has not been utilized by the aquatic flora.
In a small stream system such as the Red Cedar River the flow in marry
of the tributary streams is less than one cubic foot of water per second
except during periods of rainfall and the periphyton community and
rooted vegetation of the tributary removes the excess nutrients from
the water before it reaches the main stream. The relationship between
the tributary waters and the nutrient content of the water in the study
area could be detected only during periods of heavy run-off and during
the winter months when the biological activity was low. The nutrient-
rich water received from (the tributary streams during periods of heavy
rainfall also carries a‘ heavy sediment load which, as will be demonstrated
later, destroys the standing crap of periphyton. Therefore, the bulk
of these nutrients cannot be utilized by the aquatic flora and are car-
ried downstream without producing a biologcal response.

The highest N-N02 + N03 value, b.12 mg 1" , was recorded on July 5,

1957 when the Red Cedar River was above flood stage. When the water

\J'k
\J'l

level receded, the nitrogen value dropped to 0.2 mg l". .

The month of July 1958 provided an excellent period to study the
relationship between precipitation, stream flow, and oxidized nitrogen
in solution in the river (Fig. lb). The stream flow and precipitation
data were recorded at the East Lansing Stations by the U. S. Geological
Survey and the U. 5. Weather Bureau while the nitrogen values were de-
termined on water sanples taken at Station -0.2 of the study area.

The river was nearly at base flow prior to the rains on July 3,

h, and S which caused the water level to rise slightly over four feet
at Station -0.2. Since the watershed area was very dry prior to the
rains, the surface run-off period was very short and the river level
dropped 23 inches between July 5 and July 7, and 13 inches between
July 7 and July 9. t

The N'N02 + N03 in the river increased from 0.11 mg 1" before the _
rainfall period to 2.70 mg 1" during the peak run-off period. It then
decreased rapidly as the run-off declined. The nitrogen value was only
0.29 mg l" on July 23 and 0.06 mg 1" on July 31.

During the stuch' period the N-N02 + N03 content of the Red Cedar
River did not show a significant increase after the introduction of the
Williamston sewage treatment plant effluent even though as much as 1.2
mg 1" of nitrogen in the form of ammonia was measured at Station +0.1
in June 1958. This would appear to indicate that even thong: nitrifica-
tion may occur, the utilization rate of ammonia by the aquatic flora of
the stream is sufficiently high to prevent accumulation of the oxidized
forms.

One might question why nitrification and an accumulation of nitrOgen

in the oxidized forms do not occur during the winter months when the

 

 

Inches
Precipitation

Discharge at East Lansing in c.f.s.

’9'

mg N°N03 1"

N

H

600

500

fi‘
8

‘3’

§

00

2.0

1.0

 

fl

 

 

 

 

 

 

l
A
L
t
i 4A....
\.
i
i
9 16 F3 26 2% 50
July 1958

Fig. 114. Relationship between precipitation, stream flow, and inorganic

nitrogen in the Red Cedar River.

 

 

 

57

periphyton production was low. First, the rate of mineralization of
nitrOgeneous waste in the settling basin of the sewage treatment plant
proceeds much more Slowly during cold weather and the production and
release of ammonia was less than 50 percent of the summer values.
Second, the activity of the organisms responsible for the nitrification
of ammonia would be reduced by the subOptimal temperatures so that the
increase would occur further downstream from the outfall. And third,

in a lotic environment the ammonia introduced is carried downstream as
NHHOH or as Nfii either as the free ion or as the ion adsorbed on organic
particles so it does not accumulate in a given area and undergo nitrifica-
tion.

During the winter period the N°N02 + N03 values were slightly higher
at the downstream stations than at those located upstream from the sewage
treatment plant. The buildsup occurred when the downstream stations
were covered with ice and the photosynthetic activity was low. The
N'N02 + N03 values at Station +3.7 were greater than 1.0 mg I' on the
sampling dates between Nbvember 19, 1957 and March 26, 1958, but values
of less than 0.5 mg I“ were found after April 22, 1958.

The data indicate that the N'N02 + N03 values in the Red Cedar
River are highest during the winter months when the stream is covered
‘with ice and periphyton production is low. After the spring thaw the
periphyton production increases and the nitrOgen content of the water
decreases. During the summer months the available nitrogen content of
the water may increase temporarily as the result of precipitation and
runpoff from the watershed area, but concentration in solution decreases

Sharply after the peak run-off period. During the periods of high

U1
CU

periphyton production the total inorganic nitroEen content of the

water was frequently less than 0.1 mg I”.

Periphyton Production

 

The periphyton occupies a very important position in the energy
transfer systems operating within a lotic environment. Because euplank-
ton is virtually absent from all but the more sluggish streams, the
role of converting solar energy to organic material is carried out al-
most entirely by these forms. Occasionally rooted plants such as

Wiflisneria s . or Potamogeton s2. are found anchored to the stream bed,

 

but these forms are usually of minor importance because of their season-
al occurrence, limited distribution, and inability to recover quickly
from adverse conditions. The periphyton community is ubiquitous, and
although seasonal variations in composition and productivity do occur,
the quantities produced are sizeable at all times of the year because

of its short turnover period. Besides its position as the most impor-
tant primary producer in a stream, the periphyton mat also provides
shelter for numerous forms of benthic organisms. The herbivores could
be expected to be found in this "green pasture", but the fauna also
includes various filter feeding forms and carnivores.

Although the l.h square decimater plexiglass substrata used to
measure the periphyton production in the Red Cedar River presented a
smoother growing surface for the organisms than is naturally found in
a stream, the growth patterns on the substrata appeared to be comparable
to that on the stream bed. In other words the phytOpigment density on

the substrata was high when the periphyton pOpulation on natural submerged

 

59

objects appeared to be adding new growth. Likewise, when the flora on
the natural substratum appeared to be in a static state, the production
as measured on the plexiglass plates was low.

Qualitatively, the periphyton community of the Red Cedar River
consisted primarily of members of the Bacillariaceae (diatoms). These
organisms are characterized by their boxlike silica shells and yellow
pignent which tends to mask the green chlorophyll. These diatoms be-
come attached to the substratum by the whole of one surface of the cell
or by means of a mucilaginous stalk or extrusion. Taxonomically, this
is a very difficult group and since this study was undertaken to deter-
mine the relative production of organic material in the different areas,
identification was not attempted.

Seasonal Variations

The periphyton production in the Red Cedar River demonstrated dis-
tinct seasonal variations (Fig. 15) which were difficult to correlate
with the physical and chemical conditions of the stream.

The periphyton measurements were initiated August 6, 1957. The
periphyton production during the first study period from August 6 to
August 15, 1957 was relatively high with a mean phytOpigment accretion
of 78 AA 1: 10’ units per square decimater per day at Station +3.7.

The phytopigment production dropped approximately 50 percent during the
next stucbr period from August 20 to September 3, 1957 and continued to
decrease until the period of ice cover in December. This indicates a
decrease in the production of organic material through the fall period
and the absence of the characteristic autumn maxima frequently recorded

for a lotic enviromnent.

Fig. 15. .mean daily phytopigment production per square
decimater-measured at nine stations on the Red

Cedar River from August 1957 through August 1958.

 

 

   

 

 
 
   
   

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

300 r-
Mile + 4.9

200 —

lOO ~
> o
at Mile + 3.7
E5 200 -
D. '00 _ 5 Lljeftss Then
0: O - nn 5 per day
LU Mile + I .8
l— 200 —
LLJ
2 I00 _ Less than
B O % ml-S Units per dcy-l
Q Mile+ l .3

200 r
it
< lOO ~—
8 0

Mile + 0.6
”7 zoo —
85 IOO
* Less than

0‘ O _l‘5 Units per day-l
fl Mile + 0.5
E 200 *—
2'00 new 4
Z O _ fit n: 5 per oy
LLJ Mile + O.l
E 200 ~—
O
a: '00 H _}‘5 LLess lhorcij 4
B O I _ __ m 5 per oy
>— Mile - 0.2
I 200 r
O.

'00— Less than
<Zt O %_ [-—5 Units per day—al
L5] 200 FMile — 0.4

'00 ’— Less lhon ;' “ _

|O2O IOZO |O20l—IO20l lOZOl |O2OTIOZO IO2OTO2OllO2Ol|O2OlIOZO lO2O IO 20

JULY AUG SEPT OCT NOV DEC JAN FEB MAR APR MAY JUNE JULY AUG
| 9 57 I 9 5 8

61

During the winter period the ice cover on the Red Cedar River
reached a maximum depth of 11 inches at Station +3.7 and was frequently
covered with up to 6 inches of snow. This excluded a large portion of
the available light from the organisms on the substrata and reduced
primary production to the lower limits of detection possible by the
phytopigment method of measurement. It should be pointed out that the
depth of ice cover increased toward the downstream stations and this
was reflected in a decrease in phytopignent production. The increase
in the depth of the ice cover toward the downstream stations could not
be correlated with differences in water temperature or stream flow.

The periphyton production increased very rapidly following the
spring thaw and reached a maximum in April and early May. The phyto-
pignent production at the downstream station exceeded 130 units per
square decimater per day during this period. A slight decrease in
production was measured during the last half of May and then a sharp
increase occurred during the month of June. A measurable quantity of
ammonia nitrogen was found in the river at all stations during the June
surge in production and, since it had been determined that during periods
of high periphyton production the utilization rate of ammonia nitrogen
was equal to or exceeded the production of ammonia, it was concluded
that the source of this nutrient form was not indigeneous to the stream.
Although Resnick gt _a_l_. (1958) found that cyanates were relatively
stable under aerobic conditions, no other possible source of amnonia
could be found. It is therefore assumed that the cyanates released
into the river from the industrial plant lagoon at Fowlerville were

carried downstream by the current and slowly underwent hydrolysis.

62

The ammonia produced by this reaction stimulated the periphyton growth.
The greatest biological response was measured in the zone of phosphorus
regeneration.

The periphyton production was receding as the extra available nit-
rogen supply was being utilized, when, for the second straight year, a
heavy rainfall during the first week of July resulted in high water
levels and high stream turbidities. As will be described later, the
flood water conditions which existed for only a short period of time
almost completely destroyed the standing crop of periphyton that was
present immediately prior to the period of rainfall. After the turbid
conditions had subsided, the periphyton production was comparable to
the values measured during the late summer and early fall of 1957.

In summarizing the seasonal variations in periphyton production
in the Red Cedar River, the data indicate that growth and reproduction
increase rapidly following the period of ice cover and reach a maximum
in April or early May. If the period when the extraneous nitrOgen sup~
ply is ignored, it would appear that the production would slowly decrease
through the summer and early fall until the period of ice cover when
it is sharply reduced.

variations between Stations

The phytOpigment data indicate a discrete difference in the peri-
gflryton production between the stations located above and below the Wil-
ILiamston sewage treatment plant outfall, but the area of greatest bio-
10gical response to the nutrients acquired from the sewage treatment
plant effluent tends to shift with water temperature, stream velocity,

auui'basic productivity levels (Fig. 15; Appendix C).

63

During the August to December period when the productivity was
decreasing with each periphyton measurement period, the measured
differences in production between stations became less pronounced.

During the period from November 26 to December 10 the difference in
organic material accumulation on the artificial substrata at the upstream
and downstream stations showed very little difference. After the ice
had formed on the river, no attempt was made to compare periphyton pro-
duction and nutrient levels at the different stations. The data given
in Appendix C illustrate the low level of growth during the winter
months.

During the first periphyton collecting period after the ice period
from March 7 to March 20, 1958, the artificial substrata were exposed
only at Stations -O.h, -0.2, and +1.8 because of problems Vof accessibility.
The mean phytOpigment accumulation for each station was 631, M5, and
1,222 units, respectively. The decrease in production at the second
station was attributed to interference by turbid waters entering from
Deer Creek whose confluence with the river is 0.1 mile upstream. This
effect was noted during several periods throughout the course of the
stuchr and will be discussed in a later section.

The seasonal variations in periphyton production at the different
stations are illustrated in Figure 16. In the spring when the water
temperatures are relatively low, a significant response in periphyton
growth was noted at the station 0.3 mile dox-mstream from the outfall.
Although the production level decreased towards Station +0.6, it fre-
quently showed an increase further downstream. As the water temperatures

increased, the area of greatest bi010gica1 response to the acquired

. .34....«950 pndap pzogmwgp mwmzom 23 BOHGQ Ucw 9»an Ulcpmcoa wcoflympm
30,5 wedges ceflooaaoo wig Ed new. («on acaccficmv 98.2% had mpwcu pacifiwgofnna cwnz .3“ .wfia
HHdeSO SCAM @932
m s m N a c

 

 

I 11 ‘1 4 1] ‘ 4 d ‘ 4 ...‘.......u...wl.... ..M....

8
' 1

. DON

 

Q
(J
(*3

83 .8 .fis - a ..aé -l..

mama. «0,. us: I ma an, I .....-

mmma .NH awe . m ans -.-;.
mmma .NH Henge u m Hates - ---

ammfi .OH naposoo . om hangspasw -.::. . cos
cashew magmcmxm mpmapmnzn

 

 

pioi‘lx : we at;

if?

v

SlIUn aue‘

H.

.1";

'9
¢

1-

.flhp

65

nutrients tended to shift downstream and it appeared that the sewage
treatment plant effluent had an antagonistic effect upon the periphyton
at Stations +0.1, +0.3, and +0.6. This is difficult to explain since
no toxic industrial wastes are released into the river. and heterotrOphic
organisms are not present in great enough Immbers to compete for space
with the periphyton on the substrata at the stations just downstream
from the outfall. Also, Bartsch and Allen (1957‘) and others have re-
ported that tremendous plankton pOpulations are present in waste stabi—
lization ponds where the medium is pure sewage in various stages of
carbonization and nitrification. Although Brinely (191:2) and Butcher
(1924?) reported a suppression of stream flora by sewage, the effects
can be attributed to industrial wastes. Lackey (1956) found a sharp
reduction in the plankton population of Lytle Creek after the intro-
duction of sewage treatment plant effluent, but in this study the volume
of effluent exceeded the stream flow and septic conditions resulted.

At Station +0.3 of the Red Cedar River none of the above conditions
exist, but the periphyton production showed a decrease during the sum-
xmer months.

H The relationship between the point of enrichment and the area of
maximum biological response appears to be the' result of interactions
between several physical and chemical factors. The shift in the zone
of highest periphyton production from Station +0.3 to the areas further
downstream might be considered to be related to the water temperature.
If this were true, the acne would move back upstream in the fall as the
days became shorter and the water temperature decreased. This did not

occur during the study period; in fact, the differences in the mean

66

phytOpigment density measured at the different stations became less
apparent as the fall progressed. This is demonstrated in Figure 16
when the mean water temperatures for the September 26—0ctober 10, 1957
and the May 5-12 periods were approximately the same.

The decrease in periphyton production in the area between the
sewage treatment plant outfall and Station *0.6 is not related to the
phosphorus and nitrogen levels in the stream because the source is
constant and the level of enrichment usually increases during the sum-
mer as the dilution factor decreases. Also, the nutrients in solution
are utilized within the first 0.6 mile downstream from the outfall even
during periods of low production. This indicates the periphyton com-
munity of the area is active and that the downstream response is not
due to non-utilized nutrients originating from the sewage treatment
plant outfall and flowing directly to the lower sections.

011 the basis of the available data it is presumed that a substance
or substances formed in the primary settling basin of the sewage treat-
ment plant during the warm summer months inhibits plant reproduction
in the stream down through Station *0.6. Although the periphyton growth
and reproduction in this area is inhibited, the organisms present con-
tinue to remove the nutrient elements from the water and store them _,
at levels far above their normal requirements. After death and detach-
went from the substratum, the cells are carried downstream by the cur-
rent and undergo decomposition. The nutrient constituents are released
.to solution and produce an increase in periphyton production at the
downstream stations. The nutrient supply during the early part of the

year is also augnented by nitrates introduced with seepage and ground

water.

67

During the fall period when the water temperatures are decreasing
the antagonistic or inhibitory effect of the sewage treatment plant efflu-
ent on the periphyton pOpulation should also be reduced. This would allow
a maximmn periphyton growth in the areas justdownstream from the outfall.
A comparison of the spring and fall periphyton production periods having
nearly equal water temperatures indicates that the fall production is much
less than those measured in the spring even though the nutrient levels were
comparable. A study of the "normal" solar radiation per square centimeter
received by the area indicates that the fall values are only 75 percent of
the spring values (computed from Crabb, 1950). Therefore, it may be as-
sumed that even though other conditions may be equal, the flora of the
stream cannot respond to the additional nutrients received from the sewage
treatment plant under the lower light conditions.

The combination of the physical and chemical factors which produced
the differences in periphyton production at the individual stations was
the most obvious during June 1958 when the extraneous nitrogen supply
became available to the flora of the study area. ,

The results of the chemical analyses of the water samples taken on
May 27, 1958 indicated that the dissolved phosphorus concentration at Sta-
tions +1.8, +3.7, and +h.9 were 59, 68, and 71 pg 1"I , respectively. The
value from Station -0.h on this date was 23 pg 1'". The total inorganic ni-
trogen content of the water at all the above stations was less than 0.1 mg l".

The increase in available nitrOgen during June produced a biological
response that roughly paralled the increase of regenerated dissolved phos-
phorus in solution toward the downstream stations (Fig. 15). During this
period both nutrients were present in conCentration above the minimal levels

and the physical conditions were such that the periphyton could respond

accordingly.

 

Nutrient response

It is usually difficult to detect limiting nutrient factors in a
natural body of water and the Red Cedar River proved to be no exception.
First, it is impossible to measure quantitatively only those nutrient
compounds which are available to the plants. This is especially true
in the case of a stream receiving nutrients in the form of sewage
treatment plant effluent. The aquatic flora can utilize complex in-
organic nutrient compounds as well as many organic forms. Chu (l9h6)
reported that although perphosphate is not as good a phosphorus source
as orthophosphate, it will promote growth. In the same study he found
that the calcium or magnesium salt of inositol hexaphosphate (phytin)
'will support algae as well as orthOphosphate. No data could be found
concerning the status of the immediate availability to plants of the
phosphorus in household detergents. In the routine method for phos-
‘phorus analysis either the orthophosphate only is measured or the sample
is digested so that all of the phosphorus in phosphorus-bearing compounds
is converted to the ortho- form for colorimetric detection. In a lake
all of the forms of phosphorus are either immediately or potentially
available to the flora but in a stream the potentially available forms
should not be considered in a study area because they are carried down-
streantbw the current before they are converted to an available form
by biological or chemical action .

The aquatic flora can utilize also many forms of organic nitIOgen
whixfli are not detected in the usual ammonia, nitrite, and nitrate-
nitrOgen analysis. Ludwig (1938) demonstrated that the green alga,

ChlorellgzJ can utilize organic nitrogen in the form of many of the

69

amino acids but cannot assimilate various azo compounds. Since it

is known that the organic nitrogen content of sewage treatment plant
effluent is very high and that nitrogen is a constituent of many com-
plex compounds, the measurement of only that nitrogen which is immedi-
ately available to the flora of the stream is impossible.

Second, as pointed out by lflxipple (19118), we are measuring only
those nutrients which have not been used by the flora of the stream.
The data of Kofoid (1903) indicate that the maximum plankton production
in the Illinois River followed the highest nitrate values. In addition,
it is well established that the inorganic nutrient content of a lake
may approach the lower limits of detection at the time of a plankton
bloom as a result of the available phosphorus and nitrogen being in-
corporated into living plants. Therefore, with a "cause and effect"
phenomena controlling the nutrient levels, the chemical data must be
interpreted very carefully before assuming that a particular element
is a limiting factor in aquatic production.

The chemical data indicate that the amount of soluble phosphorus
present at all but two stations in the study area was less than the
minimal levels for maximum algal growth (Chu, 191.113) (Fig. 17).
Assmning that antagonistic factors were present at Mile +0.1 and +0.3
during the summer period, one might conclude that phosphorus was the
limiting factor for periphyton growth in the Red Cedar River. The pro-
duction data for the entire study period tend to indicate that," Other
factors are involved and although the soluble phosphorus levels are
lower than the minimums given by Chu (op. cit.) , they still exceed the
concentrations in most unpolluted waters (Juday and Birge, 1931).

The inorganic nitrogen levels in the Red Cedar River for the July.

1957 through July 1958 study period are given in Figure 18.

Fig. 17. Dissolved and sestonic phosphorus content of the

water at eight stations on the Red Cedar River.

_ Totol Phosphorus
Dissolved Phosphorus

Sestonic Phosphorus

 

 

200_ Mile+4.9

 

 

    

 

 

220 3 3 5 l
JULY AUG SEPT OCT NOV DEC JAN FEB MAZR APR MAZY7 JZUNE JULY

I958
I957 SAMPLING DATES

Fig. 18. Inorganic nitrogen content of the water at eight

stations on the Red Cedar River.

 

Nitrogen os Nitrite + Nitrate

    
 
       

Nitrogen os Ammonia

E"

MILLIGRAMS PER LITER
— ro
' b

5 I2 27 2 E3:
JULY AUG SEPT OCT NOV DEC JAN TEES MAR APR MAY JUNE JULY

957 SAMPLTNG DATES 958

 

 

72

The analyses of the periphyton production-nutrient level relation-
ship for the individual study periods (Appendix C) indicate that avail-
able nitrogen may be of more importance as a limiting nutrient factor
than the phosphorus. During the summer months when the water temper-
atures exceeded 200 C., the zone of phosphorus regeneration shifted up-
stream into the study area. This resulted in an increase in the dis~
solved phosphorus concentration in the water at Stations +3.7 and +h.9.
Although the periphyton production usually responded to the higher
nutrient levels, the increase in productivity was not sufficient to
utilize the extra phosphorus that was available. Only during June 1958

when an extraneous nitrogen supply was introduced into the river did
the periphyton produced at the downstream stations show a response
adequate to utilize the regenerated phosphorus.

During several of the study periods an increase in periphyton pro-
duction was accompanied by a decrease in the inorganic nitrOgen content
of the water. Furthermore, it was frequently noted that an increase
in the available nitrOgen supply was accompanied by a decrease in the
dissolved phosphorus. The latter phenomenon was especially apparent
at Station -0.2, downstream from the confluence with Deer Creek.

. A comparison of the total phytopigment production and the mean
nutrient levels for the 1958 portion of the study are given in Figure 19.
The data indicate the necessity for considering all of the abiotic
factors influencing the biodynamics of a stream.

Although the total phytopigment production more closely follows
the phosphorus values than the nitrogen values, the rate of phosphorus

uptake is not lineally related to the weight of periphyton produced.

73

.cogosooam oophnmflhoo Ho Hgoa opoooo mama
mafia-Be coho» an as .md «nomad 3 mmmfi .8 none: noun voice 23 won pounce?“ menace goodness
candies 59: 23. use Sausage Ho nope-:63 encode woo nag poo-Raga no coda—ages Hence

33:0 393 med“:

£33 .3000 com 23 so 3033»

.ma .mae

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

m u m o
e
i r
ooooal “man u
.a men a
.... "minim
m A m 4 am
W. I a?
n OOONH . it B
.............. x m
a ....... , .,
\\\ A
9 . ... .....ML“..4.W ......................................................... m o;
m \Vtt ,, n . ........
a 89:. \\ ,, w.
\\\\\\ I
i om
083 .
A..." was nanosecond cordon»? H38. one: u --- i 8
80m: Aha H .H was demos“: 35935” .953 532 .. .

 

 

 

cam art-wane

7h

11:. organic phosphorus :phytopigment ratio data given previously indica-
ted that the flora in the areas innediately downstream from the sewage
treatment plant outfall were removing more phosphorus per unit of organic
matter than the flora either upstream from the outfall or in the down-
stream areas. Lund (1950) found that the phosphorus content of algal
cells may vary by as much as a factor of 70. Harvey (1937) points out
that large quantities of ironFand phosphorus become adsorbed on the
surface of diatoms. A portion of this adsorbed phosphorus is taken in-
to the cell by the acidic protOplasm. Goldberg gt _a_l_. (1950) found
that as high as 50 percent of the phosphorus adsorbed on the surface

of diatoms could be washed off with sea water and termed this phosphorus
fraction as water-labile. Therefore, the quantity of phosphorus removed
from the water by chem-adsorption and intracellular metabolism of peri-
phyton might be very great in preportion to the amount of organic matter
produced. Likewise, Gerloff and Skoog (1951;) demonstrated that the
nitrogen content of a blue-green alga could be more than doubled by
increasing the nitrogen content of the media.

It follows then that a change in environmental conditions that
retard growth might not necessarily result in an appreciable decrease
in the rate of nutrient removal from the stream. Also, since the
nutrient measurements involve only the excess elements that are not
utilized by the aquatic flora, the element present in solution in the
lowest relative concentration might be the most important as a limiting
factor for periphyton production.

The decrease in phytopigment production from Station -0.h to -0.2

is assisted to result from adverse physical conditions which frequently

75

occurred at the latter station due to the Deer Creek waters which

enter the stream midway between the two areas. During periods of
rainfall the tributary stream is quite turbid and, as will be discussed
later, suspended solids proved to be very destructive to the standing
crop of periphyton.

The decrease in mean phosphorus between the two areas may have been
the result of the dilution effects by the tributary waters which were
very low in phosphorus during normal flow periods.

Station +0.1 was not considered in Figure 19 because the effluent
is not completely mixed with the river water in this area and the
chemical data are not truly representative of the water to which the
substrata were exposed. During the summer months there was also some

interference by the sewage bacterium Sphaerotilus s2.

 

The periphyton at Station +0.3 did not respond to the increased
available nutrients during the summer months. The rate of nutrient
removal from the water apparently continued to remain high even though
the periphyton growth was inhibited. This is indicated by the decrease
in the quantity of the two essential elements between Stations *0.3 and
+0.6. This may be explained in part by the previous discussion on
nutrient storage and adsorption and the data on the organic phosphorus:
phytopigment ratio at the different areas within the study section.

The decrease in the phytopigment production at Station +0.6 parallels
the decrease in the available nutrients of the station. Since both the
phosphorus and nitrogen decrease so sharply, it is difficult to deter-
mine which might be termed the limiting factor. hiring two of the study

‘periods, May 5-12 and.May 12-19, the highest phytOpigment production

76

was measured at this station. An examination of the chemical data from
samples taken April 30, May 12, and May 19 indicate the dissolved
phosphorus content of the water was abnormally high on May 12 (814 pg 1")
but on the other sampling dates it was ”normal" for the area. No dif-
ferences were detected in the inorganic nitrogen concentration during
this period.

After May 19, 1958, the production and available nutrient values
were low at Station +0.6 and the greatest chemical changes and peri-
phyton production were measured in the wees further downstream.

In the area downstream from Station +0.6 the phosphorus and phyto-
pignent values increase and the nitrogen values show a slight decrease.
If the biological response at the downstream stations was entirely due
to the increase in available phosphorus, it appears that the increase
in production would be sufficiently great to prevent the increase in
the quantity of dissolved phosphorus in solution. This would seem to
indicate that the decrease in the mean inorganic nitrogen values towards
Station +h.9 results not the increase in plant growth, but that the
quantity of available nitrogen is not sufficient to support a periphyton
population of sufficient magnitude to utilize the excess phosphorus.

The fate of the nitrogeneous compounds in the decomposing, detached
organic material from the upstream areas is unknown. The inorganic
nitrogen values did not increase in proportion to the phosphorus regen-
eration. In a previous discussion the storage and adsorption of phos-
phorus by algae were mentioned as being responsible for the atypical
organic phosphorus :organic matter ratio at Station +0. 3. It is assumed
that the quantity of nitrogeneous material undergoing nitrification in

the decomposing periphyton was not great enough to exceed the biological

77

uptake in the downstream areas and produce a measurable excess.

The maximum biological response in the downstream areas chiefly
responsible fer the increase in production (Figure 19) occurred during
two periods when the available nitrOgen was high. During one of the
periods (April 1958) the nitrogen in the stream was being replenished
by runroff water from.the drainage area and in the other (June 1958)
the available nitrogen was originating from an extraneous source. The
available nitrogensphytopigment relationship for the April 18-25 period
is given in Figure 20. The total dissolved phosphorus values for
April 22, 1958 only increased 10 pg 1"| between Stations +0.6 and +h.9.

In conclusion the data indicated a greater periphyton growth re-
sponse to high nitrogen levels than to high phosphorus levels. During
the summer months when a large amount of nitrOgen is organically come
bined in terrestrial plants in the watershed area, the stream.deve10ped
a nitrogen deficiency which inhibited periphyton growth and resulted
in an increase in the dissolved phosphorus content of the water.

Effects of Turbidity on Stream.Peripnyton

Turbid water conditions were considered as a factor which limited
periphyton production on the Red Cedar River.- The two stations were
established upstream from.the sewage treatment outfall to measure the
effects of turbidity introduced into the stream by the Deer Creek water
on the standing crap and growth rate of periphyton. The area upstream
from Deer Creek (Station -0.h) was subjected to adverse conditions only
after relatively heavy rains whereas the area downstream from the COD?
fluence (Station ~0.2) was subjected to adverse conditions following

each rainfall of sufficient quantity to produce run-off. The seasonal

78

amp 92mm anemtdoqxng ween

.cowhooooam cophnaweom mo Ho>oH epocmo mhmm
.mmma .mwuma an: new weapon on» now sebum Mateo mom on» :0 30.33» games no nonsense
330393 psesmfiooabfi one. one have: 05 we possess demand. 0.35.35 23 «o 33.39.80 .8 .wwh
damage 89C and“:

’ L ’

 

> r

o ... ta ) .1...

- 5.3.1. ‘ Mn.

m a m a H o

 

 

 

 

 

 

 

 

.....

.....
.....

 

 

 

 

 

 

 

 

 

 

ooom . somehfluz I ll

 

m
e

,1 Son a» 301.1 4 Wm.“ 3H

 

 

79

reduction in periphyton “production at the station downstream from Deer
Creek has been discussed in a previous section and illustrated in
Figure 19.

The heavy rainfall of July 3, h, and S, 1958 resulted in a four-
foot increase in water level and turbidities which exceeded 150 units.
Because of the extremely dry watershed, these adverse conditions
existed on the stream for less than a week.

By chance the artificial substrata used to measure the periphyton
production had been exposed for seven days prior to the rainfall period
and were scheduled for removal on July 3. The substrate at Station “1.9
were removed from the stream on July 3 Just prior to the rains and those
from the remaining eight stations were allowed to remain in the stream
for an additional seven days. This resulted in the units from Station
-0.h to +3.7 being exposed to stable water conditions for seven days
which allowed for the establishment of a standing crop of periphyton on
substrata. They were then exposed to approximately four days of severe
adverse physical conditions which consisted of high stream velocity and
high turbidity. This was followed by approximately three days when
conditions were returning to "nomal" (Fig. 11;). This sequence of con-
ditions provided data that clearly demonstrated the effects of turbid
water conditions on the standing crop and growth of the periphyton within
the study area of the Red Cedar River.

The measured phytopigment accumulation per square. decimater of
substratum from the units removed after seven days exposure to stable
water conditions at Station +h.9 were used to calculate the standing

crap of periphyton present at the remaining eight stations prior to

80

high-water conditions. The production relationship between the stations
was based upon the data obtained for the June 18-25 periphyton period.
The calculated density for the entire lh-day period if normal stream
conditions had persisted was estimated.by multiplying the one-week
densities by two. No information concerning the periphyton growth
curve is available, but the values are thought to be within reason.

The substrate that were allowed to remain at the eight stations
for the 1b days which encompassed the adverse physical conditions were
removed from the stream on July 10, 1958. The phytopigments were exp
tracted from the periphyton and the mean densities determined for each
station. The measured values for the 7-day period for Station th.9 and
the measured values for the lit-day period from the remaining eight
stations are plotted with the calculated.values expected if stable
water conditions had persisted in Figure 21.

The data indicate that the high water level conditions and asso-
ciated turbidities which were present during July h to July 8, 1958
almost completely destroyed the standing crop of periphyton of the
study area. The phytOpigment density of the periphyton extract from
the artificial substrate that were allowed to remain in the stream for
the entire 1h-day period can be considered to be from.growth that took
place between the time the turbidity had.subsided and the substrate
were removed from the river.

From this observation it is concluded that highdwater levels and
associated adverse conditions which may exist for only a short period
of time can, through the abrasive action of the particles in suspension
.and exclusion of light, effectively destroy the standing crop of peri-

phyton in a stream.

81

.nosam havoc pom one :H masseuse» mmmawfixmad so mcwzosm sopznmauoa we done
meanness one so one sodeospond zHoEHAQ so hpwoanasp one hopes sue: me museums consumes one enemasoamo .Hm .mwh
Hasupso scan mode:
m a m N H m

- ’ ’ )

 

‘IIOIOIO'O'CIO'O’

ammo 4H

 

 

\ when: someone: u

x 32: eeeeaeeeeo - i... use ---

 

H
z-um Satan anwfitdom Imam

é

.. 80m

 

 

Spectrophotometric Analyses of the Phytopigments

The pigments extracted from periphyton with 95 percent ethanol
are complex in nature and contain not only the chlorOphyll group but
also the xanthophylls, carotinoids, etc., and the alcohol soluble
fats and oils. Having noticed a change in apparent color of'the pigment
complex from the different stations in the study area, it was theorized
that the flora in the enriched areas might contain one of the minor
pigments in sufficient quantities to produce a characteristic absorption
spectrum and thus serve as a criterion for detecting organic enrichment
or stream pollution.

A qualitative periphyton sample was collected from Stations -O.h
and +0.1 during the winter, spring and summer periods, the phytopigments
extracted with 90 percent redistilled acetone, and the absorption spectra
determined on a Beckman Model "B" spectrOphotometer. The data indicate
that a qualitative difference in the absorbancy spectra of phytOpigz'wnt
extracts from the different stations could not be detected until a

growth of Sphaerotilus 32. was evident at Station +0.1. This sewage

 

bacteriumvms first observed on July 3, 1958 and was thought to be as-
sociated.with the high carbohydrate content of the sewage treatment
plant effluent due to the home canning season. It was never observed
at any of the other downstream stations during the study period.

The absorbancy peaks for a phytopigment extract complex from a
periphyton community from the stream section unmodified by sewage
treatment effluents are at h32 and 660 millimicrons. This is demonstrated
in the spectrum of phytOpigments from Station -O.h (Fig. 22). The

spectrum having absorbancy maxima at h10 and 665 millimicrons was

Absorbancy

 

------ - Station -O.h
_- Station "0.1

07’

.6 o

05‘

o
32'

.31

02‘

.1

 

 

 

 

uoo 500 600 700
Millimicrons

Fig. 22. Absorbancy spectra of the phytopigments from two
stations on the Red Cedar River.

83

obtained from phytOpigments extracted from periphyton collected at
Station +0.1 on July 23, 1958. During this period a dense growth of
heterotrOphic organisms was noted on the artificial substrata in this
area. The shift in the absorbancy maxima is attributed to the presence
of decomposition products of chlorophyll (Holt and Jacobs, 195h).

These products were formed after the heterotrOphic growth destroyed
the periphyton accumulation on the substrata.

In view of the above data it was concluded that a spectrophoto-
metric analyses of phytOpigment extracts could not be used to detect
areas of organic enrichment or pollution in a stream until the pOpu-
lation of heterotrophic organisms became sufficiently high to destroy

the autotrOphic community.
Proggpted Intercommunity Relationships

A complete study of the biodynamics of a lotic environment would
involve'uueanalyses of the physical and chemical factors which control
the biological production plus a quantitative measurement of the bio-
logical production of each trophic level. Because of time limitations,
the chemical analyses were confined to determining the phosphorus and
nitrogen content of the water and the biological study involved only
those organisms most closely dependent upon these nutrient elements.

Since Charles Elton first prOposed the pyramid of numbers, the
relationship between the higher biotic units and the primary producers
has been studied and substantiated in both terrestrial and aquatic en-
'vironments. Lindeman (19b2) proposed the trOphic-dynamic concept of

ecology and stated that the caloric content of the organisms composing

85

each trophic level was homologous to the pyramid of numbers. Therefore,
in view of the intensive and extensive work indicating a biological
response of the higher organisms to increases in primary production,

the scope of this investigation was limited to the basic producing units
in the environment and the data used as an index to the production within
the higher trophic levels.

The relatively short section of stream involved in the study above
the sewage treatment plant outfall (0.h mile) presented two distinct
types of habitat. In the area below the Deer Creek confluence, the
organisms were subjected to periodic, highly adverse conditions during
highdwater stages. The flood waters from Deer Creek arise in part from
cultivated land and are highly turbid and carry a heavy bed load of
sediments. The fine particles in suspension were detrimental to the
flora and fauna in that they exclude light from.the former, and the
larger particles (chiefly fine grains of sand) mechanically destroy
the biota by their abrasive action. The primary producers are able to
recover rapidly after conditions return to normal but the primary
consumer papulation would probably remain low until recolonisation
was affected by drift from.upstream areas or ovaposition by adults
which had emerged from other areas. Therefore, the reduction in the
measured primary production at Station -O.2 might well be indicative
of a reduced papulation of benthic forms, not only as the result of
less available food, but also as the result of adverse physical con-
ditions.

Although the flora at Station -0.h was subjected.to periods of

turbid water conditions, most of the coarser silt particles and sand

86

settle out in the upstream reservoir. It has been demonstrated that
the periphyton in this area recovered more rapidly than at other areas
following high-water conditions and it can be assumed that the benthic
forms here were subjected to less adverse conditions.

It would be impossible to project the periphyton data to the
fish population in the stretch of river between the sewage treatment
plant outfall and the Williamston Dam because of the short distance
involved and the ability of fish to migrate from areas of less favorable
environmental conditions to areas of more favorable conditions. In
addition, the dam acts as a barrier to upstream migrants and undoubtedly
served to concentrate those species attempting to move upstream in the
Station -0.h portion of the area.

The flora in the immediate area downstream from the sewage treat-
ment plant outfall tends to store the nutrient elements in excess of
their basic requirements and this produces an intermediate area where,
during certain periods of the year, the measurable nutrients in solution
are comparable to or less than the levels upstream from the outfall.
This raises the question as to whether the production of organisms in
the higher trophic levels would increase at Station +0.3 and decrease
again at Station +0.6. In reviewing the data it can be seen that the
zone of maximum periphyton response shifted from the Station +0.3 mile
area in early spring to’the downstream areas during May and June. Since
the benthic forms could not migrate with the shift in maximum peri-
phyton production, it is assumed that the population would be controlled
by the minimal conditions in the habitat. This was confirmed in an

earlier study (Brehmer, 1956) in which the benthic association in an

87

area downstream.from a point of organic enrichment was composed of
fewer taxonomic groups than at other areas in the stream even though
recognized adverse conditions did not exist.

The increase in total periphyton production in the downstream
areas is significant in many respects. Butcher (19h?) noted a sharp
decrease in sessile organisms immediately below the point of pollution
but the pollutants included tar acids, gas liquors, and organic Chemical
wastes which would suppress the pOpulation. In his study the first
population maximum was measured eight miles downstream from.the sewage
outfalls. 0n the Red Cedar River the maximum production was measured
at Stations +3.7 or +h.9 only during 8 of the 2b study periods from
August 6, 1957 to August 15, 1958. The point most important in consider-
ing the relationship of periphyton production to the higher biotic
forms is that the production in the downstream part of the study area
'was always relatively high during the summer months when the activity
of the poikilothermic herbivors is the greatest. There were no periods
when the daily periphyton production dropped to almost zero as was noted
in the areas within a mile of the outfall. Also, the maximum periphyton
response was noted during April and May, a period of the year when the
'benthic forms are in their final instars prior to emergence and.the
grazing rate is high. The biological characteristics of the Station
+h.9 area were reported in a previous study (Brehmer, 22. £22.). The
‘benthic association was not only more complex in that a larger number
of taxonomic groups were represented.but also more of the forms present
'were in the groups considered.to be more suitable and.available as fish

food.

88

In view of the extensive literature relating benthic fauna to
fish production (Ball and Hayne, 1952), it would appear valid to assume
that the downstream areas are capable of supporting a greater fish pop-
ulation than either the area upstream from or immediately downstream
from the sewage treatment plant outfall. Katz and Gaufin (1952) found
that not only the total number of species and individuals of fish in-
creased with distance from the sewage outfall, but that the fish failed
to move upstream towards the outfall during the winter months when the
dissolved oxygen content of the water became tolerable. The data from
this study indicate that even though septic conditions did not exist
at Stations +0.1, +0.3, and +0.6, the fish food organisms might be
limited by periods of very low periphyton production.

Therefore, if the fish and bottom fauna production of an area
are related to the minimal periods of periphyton production, a study
of this type using artificial substrata and the phytOpigment density
of the accumulated.periphyton as an index might well serve to classify

a stream or an area thereof as to its recreational potential.

SUMMARY

w \le!!!)fx ‘\1l{/\!I\\/)\ ‘

SUMMARY

1. The mean dissolved phosphorus content of the Red Cedar River ‘
at Station -0.h was found to be approximately 30 pg 1“ (range - 13 to 53).
The mean inorganic nitrogen content of the water in this area was 0.7
mg 1" (range - trace to 1.86). The quantity of nutrients in solution
varies with stream flow, water temperature, and primary production
levels.

2. The greatest influx of nutrients from the tributary streams
occurs shortly after the start of a rainstorm and the concentration de-
creases rapidly during which time the activity of the aquatic flora is
suppressed by adverse physical conditions. Thus the greatest proportion
of those nutrient elements leached and eroded from the watershed are
lost from the ecosystem without producing a biological response.

3. A major portion of the phosphorus and nitrogen available to
the aquatic flora is introduced into the Red Cedar River from municipal
drains and sewage treatment plant effluents. The phosphorus accrual
from the Williamston sewage treatment plant effluent exceeds 100 pg 1
during periods of normal stream flow. The annual phosphorus accrual
from this source is approximately 2.5 metric tons.

h. The inorganic nitrogen accrual from the Williamston sewage
treatment plant effluent exceeds 0.5 mg 1" during periods of normal

stream flow. The organic nitrogen content was not determined.

91

5. The accrued nutrients are removed from solution by biological
uptake and/ or chemical adsorption or precipitation within 0.6 mile
downstream from the outfall. During periods of high stream temperatures
and normal flow the biologically combined phosphorus is again released
to solution within the 14.9 mile study area by the decomposition of
periphyton which becomes detached from the area below the outfall.

The accrued nitrogen did not reappear in solution in the inorganic form
within the sturh' area.

6. The ratio of organic phosphorus to phytOpigment in the peri-
phyton was more than four times greater at Stations +0.3 than at -0.2.
The ratio decreased with distance from the outfall. This indicates
that the aquatic flora in the enriched area is storing phosphorus in
amounts over and above their normal requirements.

7. The data indicate that under normal conditions the periphyton
production would increase rapidly after the period of ice cover until
a maximum was reached about May 1. The rate of productivity would then
decrease gradually until the winter ice period which sharply curtails
growth. During the stuchr period an extraneous available nitrogen source
produced a second productivity peak during June. This growth pattern
might be altered from year to year by changes in the distribution of
the annual rainfall.

8. The biological response to the introduced nutrients was great-
est during periods of high production and least during periods of low
production. Variations in the depth of ice cover at the different areas
made comparisons impossible during the winter period. The point of

greatest biological response tended to shift downstream with increasing

92

water temperatures. This was also associated with an apparent antago-
nistic action of the effluent towards the periphyton at Stations +0.1
and +0.3. The measured net phytopigments for the five-month ice-free
period of 1958 indicated that the production decreased immediately be-
low the point of introduction of the effluent, increased at Station +0.3,
decreased at Station +0.6, and then increased rapidly toward Station +b.9.
The total production at Station +h.9 was more than double that measured
at Station -0.2, the first station upstream from the outfall.

9. The increase in production at Station +h.9 was accompanied by
an increase in the mean total dissolved phosphorus in solution and a
decrease in the mean inorganic nitrogen in solution. This is inter-
preted as indicating that nitrogen is the nutrient that limits peri-
phyton.production in the Red Cedar River and results in the accumulation
of dissolved phosphorus in the downstream areas. This interpretation
‘was supported during the June production peak, when, as the extraneous
nitrogen was made available to the flora, the quantity of regenerated
phosphorus in.solution decreased.

10. The data indicate that adverse physical conditions in the form of
high stream flows and accompanying high turbidities, even though they
'may last for only a short period of time, can completely destroy the
standing crop of periphyton in a river. The flora quickly becomes re-

established as stream conditions return to normal.

LITERATURE CITED

LITERATURE CITED

Allen, K. R. 1951. The Horokiwi Stream. New Zealand Marine Dept.
Fisheries Bull. No. 10, 231 pp.

Alexander, G. R. 1956. The fertilization of a.mar1 lake. (Unpub.
'Master's thesis, Michigan State Univ. Library).

APHA, AHWA, FSIWA. 1955. Standard.methods for the examination of
water, sewage, and industrial wastes. 10th Ed. Baltimore,
waverly Press Inc. 522 pp. '

Ball, R. C., and D.'H; Kayne. 1952. Effects of the removal of the
fish population on the fish-food organisms of a lake. Ecology,

3hl‘h80

Barrett, P. H. 1953. Relationships between alkalinity and adsorption
and regeneration of added phosphorus in fertilized.trout lakes.
Trans. Amer. Fish. Soc., 82: 78-90.

Bartsch, A. F., and M. 0. Allum. 1957. Biological factors in treatment
of raw sewage in artificial ponds. Limnol. and Oceanogr., 2:

77'8he

Blum, J. L. 1957. An ecological study of the algae of the Saline
River, Michigan. HydrobiOIOgia, 9: 361-h08.

Brinely, F. J. l9h2. The effects of pollution.upon the plankton
population of the White River, Indiana. Invest. Ind. Lakes and
Stremns, 2 : 137-1143.

Brehmer, M; L. 1956. A biological and chemical survey of the Red
Cedar River in the vicinity of Williamston, Michigan. (Unpub.
Master' 8 Thesis, Midhigan State Univ. Library).

Butcher, R.'W. 1932. Studies in the ecology of rivers. II. The micro-
flora of rivers with special reference to the algae on the river
bed. Anne Bets , ’46: 813‘8610

. l9h7. Studies in the ecology of rivers. VII. The algae of
organically enriched waters. J. Ec., 35: 186-191.

Chu, S. P. l9h3. The influence of the mineral composition of the medium
on the growth of planktonic algae. Part II. The influence of the
concentration of inorganic nitrogen and phosphate phosphorus.

Jo ECe, 31: 109'1’48.

. 19146. The utilization of organic phosphorus by phytOplankton.
Jo Hare B1010 ASSOC. Us Ko, 26: 285-2950

Cooke, w. B. 1956. Colonization of artificial bare areas by micro-
organisms. Bot. Rev” 22: 613-638.

Crabb, G. A. 1950. Solar radiation investigations in Michigan. Mich.
State Agr. Exp. Sta. Tech. Bull. 222, 153 pp.

Dobie, J. , and J. Moyle. 1956. Methods used for investigating pro-
ductivity of fish-rearing ponds in Minnesota. Minn. Dept. of
Cons., Div. of Fish and Game. Spl. Pub. No. 5, 51: pp.

DobsoB, J. G. 191:7. Disposal of cyanide wastes. Metal Finishing,
5: 78-810

Einsele, W. 191.1. Die umsetzung von zugeffirtem anorganischer Phosphat
in eutrophen See und ihre Rflckwirkung auf seinen Gesamthaushalt.
Z. Fisch., 39: hO7-h88. (Cited from Biol. Abs., 23: 10136).

Eldridge, E. F. 1933. Removal of cyanide from plating room wastes.
Mich. State College Eng. Exp. Sta. Bull. 52, 20 pp.

Ellis, M. M., B. A. Westfall, and M. D. Ellis. 19118. Determination of
water quality. U. S. Dept. Inter. , Fish and Wildlife Serv., Res.

Rept. No. 9, 122 pp.

Gerloff, G. C., and F. SkOOg. 1951:. Cell contents of nitrogen and
phosphorus as a measure of their availability for growth of
Microcystis aerugcinosa. Ecology, 35: 3h8-353.

Goldberg, E. G. , T. J. Walker, and A. Whisenand. 1951. Phosphate
utilization by diatoms. Biol. Bull. (Wood's Hole), 101: 2724-2814.

Grzenda, A. R. 1955. The bi010gical response of a trout stream to
headwater fertilisation. (Unpub. Master's thesis, Michigan
State Univ. Library).

Harvey, W. H. 193h. Measurement of phytOplankton papulation. J. Mar.
Biol. Assoc. U. K., 19: 761-773.

. 1937. The supply of iron to diatoms. J. Mar. Biol. Assoc.
U. K., 22: 205-219.

. 1910. Nitrogen and phosphorus required for the growth of
phytOplankton. J. Mar. Biol. Assoc. U. K., 21;: 115-123.

Hentschel, E. 1916. Biologische Untersuchungen fiber den tierschen
und pflansenlichen Bewuchs in Hamburger Hafen. Mitt. Zool. Mus.

Hamburg, 33: 1-172.

96

~

Hepher, B. 1958. On the dynamics of phosphorus added to fishponds in
Israel. Limnol. and Oceanogr., 3: 811-100.-

Holt, A. S., and E. E. Jacobs. 195B. Spectrosc0py of plant pigments.
Am. J. Bot., b1: 710-722.

HOOper, F. F., R. C. Ball, and D.‘W. Hayne. (ms). Effects of ferti-
lization of a marl lake.

Huntsman, A. G. 19h8. Fertility and fertilization of streams. J. Fish
Res. Board of Canada., 7: 2h8-253.

Juday, C., and Birge, E. A. 1931. A second report on the phosphorus
content of Wisconsin lake waters. Trans. Wis. Acad. Sci. Arts
Lett., 26: 353-382.

Kata, M., and.A. R. Gaufin. 1952. The effects of sewage pollution on
the fish population of a midwestern stream. Trans. Am. Fish.
Soc., 82: 156-165.

Kofoid, C. A. 1903. The plankton of the Illinois River 189h-1899,
with introductory notes upon the hydrography of the Illinois
River and its basins. Part 1. Quantitative investigations and
general results. Bull. I11. State Lab. Nat. His., Vol. 6,
Article II: 95-629.

Kreps, E., and N. Verbinskaya. 1930. Seasonal changes in the Barents
Sea. J. de Conseil., 5: 327-3h5.

Lackey, J. B. 1956. Stream enrichment and microbiota. Pub. Health
Rep., 71: 708-718 .

Lindeman, R. L. 19h2. The trophic-dynamic aspect of ecology. Ecology,
‘ 23: 39941-180

Ludwig, c. A. 1938. The availability of different forms of nitrogen
to a green alga. Am. J. Bot., 25: hh8-h58.

Lund, J. W. G. 1950. Studies on Asterionella formosa, Hass.
II. Nutrient depletion and the spring maximum. 3. Ec., 38: 15-35.

 

Manning, W. M., and R. E. Juday. 19h1. The chlorOphyll content and
productivity of some lakes in northeastern Wisconsin. Trans.
Wise Acad. SCie Arts Lett., 33: 363-393.

Moore, E. w. 1939. Graphic detemination of carbon dioxide and the
three forms of alkalinity. J. Amer. Water Works Assoc., 31: 51-66.

Needham, P. R., and R. L. Usinger. 1956. Variability in the macro-
fauna of a single riffle in Presser Creek, California, as indicated
by the Surber sampler. Hilgardia, 2h: 383-h09.

 

97

Patrich, R., M. H. Hohn, and J. H. Wallace. 1951;. A new method for
determining the pattern of the diatom flora. Notulae Natures,
259: 1-12.

Purdy, W. C. 1923. A stuchr of the pollution and natural purification
of the Ohio River. I. The plankton and related organisms. U. 5.
Put). Health 31111., NO. 13, 78 pp.

Resnick, J. 13., W. A. Moore, and M. B. Ettinger. 1958. Behavior of
cyanates in polluted water. Ind. and Eng. Chem. , 50: 71-72.

Richards, F. A., with T. G. Thompson. 1952. The estimation and
characterization of plankton populations by pigment analyses.
II. A spectrOphotometric method for the estimation of plankton
pigments. J. Mar. Res., 11: 156-172.

Rudolfs, W. 19147. PhOSphates in sewage and sludge treatment. I.
Quantities of phosphates. Sewage Works J. , 19: 143-50.

Ruttner, F. 1952. Fundamentals of limnolog. Univ. Toronto Press.
2142 pp. Translated by D. G. Frey and F. E. J. Fry.

Schmidt, E. L. 1956. Soil nitrification and nitrates in waters. Pub.
Health Rep., 71: 1497-503-

Taylor, D. M. 1937. The determination of phosphates in natural waters.
J. Am. Water Works Assoc. , 29: 1983-1991.

Whipple, G. S. 19148. The microscopy of drinking water. 14th Ed.
New York, John Wiley and Sons. 585 pp.

Whiteside, E. P., I. F. Schneider, and R. L. Cook. 1956. Soils of
MiChigane Midis State Agr. Ego-Sta. Spec. 311110 1102‘. 52 pp.

 

APPENDIX A

3O

.20

10

 

I

A L

 

‘ A A A; k A A A A - A A - g - A A A

 

 

5 10 15 2O 25

Date
Daily high and low ‘water temperatures recorded during JULY 1957 at
a Station ten miles upstream from the mouth of the Red Cedar River.

°c.

30

20

10

 

H

A A A A A A A A A A A A

 

A A A A A A A A A A

 

v

S 10 15 20 25 30
Date .

Daily high and low water temperatures recorded during AUGUST 1957 at

a Station ten miles upstream from the mouth of the Red Cedar River.

 

30

.20

°c

10

 

 

A A A A A

 

 

15 20 25 30
Date
Daily high and low water temperatures recorded during SEPTEMBER 1957 at
a Station ten miles upstream from the mouth of the Red Cedar River.

5 9' ' "1b '

OC.

n-b

 

 

 

 

H 1
1 <
41 4
3o~ ' .
1’ 1
1 4
1 t
H t
20" u
.. .
‘ d
‘1 1
10 w ' .
n .
IL
OLkAI #
S 10 15 20 25 30
Date

Daily high and low water temperatures recorded during OCTOBER 1957 at
a Station ten miles upstrean from the mouth of the Red Cedar River.

C.

A-S

 

 

 

 

p
w
30 ~
20 c
b
1v
1’
10 ..
1r
«1
d
0 u
""5 ' 1'0 '15" 20 25 3o
‘ Date

Daily high and low water temperatures recorded during NOVEMBER 1957 at
a Station ten miles upstream from the mouth of the Red Cedar River.

30

20

C.

10

A-6

 

v
A

 

 

E AA A \AA - A
'—_‘
V—v v A, A
A A A A A A A A A AA A A A A A A AA 9
A A
v v

5 10 15 20 25

Date
Daily high and low water temperatures recorded during DECEMBER 1957 at
a Station ten miles upstream from.the mouth of the Red Cedar River.

 

CC.

30

20

10

A-T

 

PERIOD OF CONTI$UOUS ICE COVER

 

 

 

 

January February

Daily high and low water temperatures recorded during JANUARY and FEBRUARY
1958 at a Station ten miles upstream from the mouth of the Red Cedar River.

0C.

 

A A A

 

 

 

15 20 25 I)
Date
Daily high and low water temperatures recorded during MARCH 1958 at
a Station ten miles upstream from the mouth of the Red Cedar River.

5 16

°c.

 

304

10*

v

 

A A A A A A A A A A A A A A A A A A A A A

 

S 10 15 20 25
Date
Daily high and low water temperatures recorded during APRIL 1958 at
a Station ten miles upstream from the mouth of the Red Cedar River.

 

A-lO

 

 

 

1}

1P

30 ’
H

D

1)

1

20 h

o'

o 4
J»

‘1

1
101
1

1

1

‘r

04

5 m ' fi' fifi'f"fi'vvvfi
Date

Daily high and low water temperatures recorded during MAY 1958 at
a Station ten miles upstream from the mouth of the Red Cedar River.

 

A-ll

 

 

0 0‘

 

A AA
v

S 10 15 20 25 30
Date

Daily high and low water temperatures recorded during JUNE 1958 at

a Station ten miles upstream from the mouth of the Red Cedar River.

°c.

A-lZ

 

 

 

 

1

1D
0
4r
30 1
4
U

4
‘T
201
1

b

1.

1D
10 1
d
0H

5 j 1'0' '1'5' ivzb VESVHf
Date

Daily high and low water temperatures recorded during JULY 1958 at
a Station ten miles upstream from the mouth of the Red Cedar River.

7

APPENDIX B

B-l

ma
hm

a.”
8m

Wad

m.

hm

mum

ON

4
M

338 53:3: «mfimfia page 3855 3&8: .ms 55 33m

oowmmo 5%.“:on .mcflmcmq atheism HGOfimoaooc .mS Scam span."

 

A ..H 93 moz + NozESoafiz

A .H m5 amzéowouflz

 

 

mm 3 an N: 8 on On mm 8 mm A 7H m5 mfiofimofi H33
m.m hm hm ad «.0 a6 «a m; we. N.m TH wishes 833mg
A333 333.5
5.5 a; ad. 0.“ mg. m.» m; mé 05 mg. ma
NE SN m3 Rm EN :3 2m 08 m5 New A .H was spandex:
A ,s ..mgmimfis
o.mm mém m.m~ 9mm m.mm m.mm mspm 9mm o.m~ o.mm Toov wépmaogoa amps:
No.3 .. 30A otnm. I swam Toov manpmuoasoa .34
mm A.u..«.ov m ed“: awhmnomg
ABA 38.3 m «is Emma owes

mumml mum... .mm mummy mammal mam. mam: magi Ami m... 585 28H 3%:

RS .3 82. .. 35m .58 e

um 05 no 3339309330 H3255 use H335

B-2

 

mH mm
0.0 «.0

o m
a.a m.~

saw gem
o.aH o.Hm
F.4m n.4m

ooneo camHaOHz .mchcma puma .saoasm umgpamz..m .s scam gamma

8.33 53332 .6:me Qw>§m Hmowmoaooc .m .3 Scan memo."

 

A .H mEV moz ... wozéowonpwz

A ..H may 432.comonpwz

so 45 mm NQH mm ma om as mm mOH A .H may mahoganosa Hasoa
m.a o.m 4.m a.m m.m m.o a.m 0.0 o.m o.o A ,H msv sewage em>HommHn
m «H mH 4H NH NH NH 0H NH 4H AmpHcsv auHertsa
m.~ m.» Q.» Q.“ a.~ a.a a.a o.w H.m o.m ma
cam mew mew gem mam cam mam mmm mmm gum A .H mew apHcHmeHa

A ..Eo 1230 .70..“ as
asH>Hsosecoo
m.HN m.om o.HN m.Hm m.Hm m.HN o.mm m.wm m.mm o.mm A.oov manpwamassa you»:
«.1: .. 20A 4.0m .- nMHm 700v 233353. .34

wow A.m.m.oV m oHHz mmaaaomHn
Hmm.m Apooav m mHHz_sawme amuse
m.Hm Hém m.m~ EH m.mH 0.3 «.9 ”#3 m; m3 583 gas 8:2

 

 

 

omma .0“ 056 .. .Hmbfim goo com 05 mo mofimwnopomamno Howe—one use .33.th

B-3

ooammo nmmwnuaznqmcfimcmq pmmm .smmpsm hogpmmz..m .: Scum mpmn

m

moammo :mmanowz .mcwmcmq «hm>hsm HmOfionomw .m .D Scum mpmna

on m: Hm am am 04m mm H: Hm Ho Hm mmH
e.m o.m 4.4 m.a m.m a.a o.m N.m o.m m.m e.m e.m
o m a m mH mH MH m m a mH MH
m.~ m.~ m.~ m.~ m.~ «.5 m.~ m.~ m.~ m.~ m.~ m.~
Now wow new mom com com gem wow wow wmm 4mm «mm
o.aH m.~H .m.mH m.wH m.aH m.mN o.mm o.- m.Hm m.Hm m.Hw m.~m
mm.mH u 30H m.mm u :me
maH
Hae.m

5.4m n.4m m.Hm H.a~ m.mm m.mH m.mH o.oH e.mH .oH m.~ m.a

 

 

A .H may moz + Noz.cwmousz
A .H wEv amz.:owoup«z

A .H may ushonmmonm dance

A .H wsv cowhxo cm>aommwn
Gaga 3483.2.

an

A .H may awwcwflmxdd

120 amaze 10H xv
apH>Hsosecou

A.oov ohsponogsoe hoom3.
A.oov wasponQan afi<
A.m.m.ov m oHHz.omaanuaHn

“gooey m 0HHz pngmm amuse

 

nvsoE_80am sea“:

omma .mw case n Ho>fim amuse com map mo moflpmwAmpomndno Heedsono use Hmowmhgm

 

0H
0.0

m
o.m

mam

0.3H

5.4m

0H
o.o

m
H.w

00m

m.mH

3...

ma

0.5

H.©

com

o.mH

m.am

mm
«.0

com

com

0.0N

H.5m

m:
0.0

H.m
mum

Ooow

m.mm

SH
m.m

oow

mom

O.Hm

w.ma

833 53842 asses swam .323 353: .m .a 22a 38m

ooamuo camanowz..mcwmcmq «ho>asm awoawoaooc .m .9 Bonn sauna

 

A ..H m5 moz + «02%mesz

A 2H wsv gmz.cmm0Asz

Ha mm aw Hm 3 me A ..H m3 mfiofimofi H38.
:6 N; do o6 o; a: A ..H m5 8906 35238
m o m m a w Ampgv .3333
o.m H.© H.w N.w ~.m H.w an
mom 48 mm“ new 08 4mm A ..H 95 .323er

A 150 wheno 70H NV
hpabwposccoo
o.Hm an un o.mm 0.4m m.4m A.oov caspanoasms noon:
~w.mH u son 4.0m u ame A.oov oasaaumaawa 9H4
mm A.m.u.ov m saw: omhmnomwn

HBA 33.: m mHHz EmHmm «mama

 

m.mH o.pH o.MH m.oa m.~ m.:

 

58: see nods

emmH .om 8% .. 35m 58 Ham 23 .3 832398.85 H3220 new 1335

_,_. ‘ AHA-A ‘.——._... .

 

oonmo :mMHao z .wcHacua swam .saossm posses: .m .: Esau «swam

003.8 H3933: «msHmcmA QQEdm Hmofimoaomo .m .3 Song. .334

 

A .H may moz + N02.53.”sz

A ...... way Jazéomgfiz

mH 0H mm 00 00 OHH amH 00 H0 mm m0 0a A .H may mauoaauogm Hence
5.0 0.0 H4. a; mé «.0 m.0 0.0 m.0 0.0 m4. 0.0 A .H was sembno 025332—
m a a m a 0 0 a m a 0 0 323 333.39
mo.m mo.m oo.w mm.~ mo.m oa.~ om.~ oa.a ca.a mo.w OH.® mo.m ma
«mm 0mm 00N 00m 00m cam cam 00m w0m 00m 00m 00m A .H mev auHcHHaxHa

A .-Eo 1250 .70.... RV
anH>Hpo=ecoo
m.0H m.0H m.0H 0.0H m.0H o.o~ m.aH m.mH o.aH m.mH o.mH o.H~ “.000 mpspathEoe topaz
n.4H -_aoH H.~H - amHm A.oov massagoasoa uHa

a» A.m.a.00 m oHHz mmnmnoan
Hoa.m Hpmmuv m wHHz pamHmm amuse
5.4m min m.Hm H.~.~ m.m~ 0.3 m.mH 0.0H 0.2 m.0H 0;. m.m 58:50am umHHz

0mmH am 32. n .823 9330 com on... go megawampomamgo H3305 use 30de

 

 

83.00 535.“: «ufinzmq puma .smmhsm 5.5303 .m .D Eon...“ wpmnm

oofiumo :mwanoflz .mcfimqmq .ampusw Haoamoaomo .m .p 20g“ mpmna

5—6

 

A 2H msv moz + moz.cmwoapfiz

A .H may aszmwonpaz

ma 0H mm 3 on 50H mma pm ..3 3 mm 00 A ..H m3 waonamosm Hmpoa
A ..H may «8906 02503.3

4 a 4 a m m m 4 a m 0 m Ampflqsv huauanasa
A ow.m mm.m m~.m mo.w mH.w mm.” mm.~ 00.x mo.m mo.w oo.m oo.m an
8“ com 8m. 08 wow OR. mom wow .03 8m 8m 08 A ..H 95 33¢an

A .-Eo 1950 ..-OH xv
agfipaposcaoo
m.ma m.om o.am m.mH m.om m.o~ . A.oov mgfipMQQQEma noamz
«0.3 .. Boa mém u swam Toov wuspmhmnsoa HQ

mp “.m.m.00 m maflz omnmgomfin

Hmm.m Apmmmv m ma“: pnmfimm mmsmo

 

 

55m min mtnm. Héw momm 0.2” méH 0.0a 0.mH m.OH 04. mu... 5:02 50am anH:

0mma .oa hash . nm>am “acme cam mg» no mafipmfluoaoupmno Hmoasugo on» Hmofimhgm

 

ooammo :mmwsoflz «wcawcdq pmmm «smwhdm hospmozv.m .D saga mawnm

mowwmo :mmwnofl: nmcamcwq .hm>nsm Hmoamoaowo .m .2 scam momma

 

A ...n MEV moz + Noz.nmmo.5.wz

A .H m5 :mz.cmwo.np.wz

gm 00 pm Hm «NH mad mma oHH am 0ma AMH now A ,H mav mauonamogm Haves

H.m ~.m 4.m «.3 . H.m .m.m m.m 0.0 m.m ~.m m.m m.m A 2H wev :mwhwo vmpaommwn

ma AH 0H Hm mm m; .om mm mm mm mm mm Ampaasv mpficfinase

mm.h om.A om.~ o:.~ om.~ oo.» 00.» m0.» m0.» 0”.” m~.~ mm.~ ma

New mam Jam wow wow oma OmH amfl cam OmH «ma 05H A AH may hpficflfluxaq
on: m0: me: am: mo: no: mm: on; Q04 mm: mas «pm A :50 uneno _voa xv

. huflpaposuaoo

o.mH o.o~ o.o~ o.mm m.o~ m.- o.m~ m.A~ o.mm o.4~ o.4~ m.m~ A.oov waspmgmqsms away:

H.0H s seq m.w~ . nMfim A.oov mgppmpmgema ““4

m0m A.m.m.ov m mHHz mmumnonwa

Ham.4 Apommv m 0H“: pnwfiom mmsmo

 

 

 

b.4nl. m.4m M.Hm H.5N m.m~ m.wH m.mH 0.0a 0.mH n.0H m.w m.: 2950: Scum mmHH:

 

 

0mma .oH pmswa< . gmpflm nuumo com on» mo,m0fipmfinopomaunu Haofiewno cam HmOfiwhgm

23-8

30.0.00 §mEo§ £00.3ch 0000M 0:00.03 .3533 .m. .2 29C. Sana

003.3 8032on «magma £05.05 Quawoaooo .m .2 50.0.0 mama—H

 

00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 A 0H 000 002 + 002.00000002
00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 A .H 000 002.00000002
00 00 00 00 00 000 000 00 H0 00 000 A .H 0:0 0000000000 00000
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 .0.0 A 10 000 0000x0 000000000
0 0 0 0 0 0 0 0 0 0 0 A000000 000000000
00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00.0 00
00m 000 000 00m 00m 000 000 000 000 000 000 A 1H 000 0000000000
20 000 000 000 000 000 000 000 000 000 000 A .000 1050 .-0H 03
. . 000000000000
m.ma 04m 0.3 mém m.m.o. mSm 0.0m o.mm 0.3 M43 0.0m 700V gpmummsms .3003
00.00 u 300 0.0m . 0000 A.000 00000000000 000

00 A.0.0.0v 0 0002 000000000
H00.0 A00000 0 000: 000000 00000

5.4m min mtnm H.5N m.m~ 0.3 M60“ 0.0a 00m...” M5.” 0.0. m3 $.90: ERG. no.3“:

 

 

0000 .00 000004 . 00000 00000 000.000 00 000000000000000 00000000 000 H0000000

 

00.0
0.0
0
0.0
0
00.0
000
000

0.00

0. m

3.0.0.00 00.09.050.000 «M00359 000m 00009005 0020.003 .m .2 08.0.0 3.009

m

30.0.00 50300002 09000ch 0.020.020. 000.000.0000 .m .2 08.0.0 000.90

00.0 00.0 0m.o 0m.o 00.0 mm.o
0.0 0.0 0.0 0.0 00.0 00.0
00 00 00 00 000 000
0.0 0.0 0.0 0.0 0.0 0.0
0 0 0 0 0 0
00.0 00.0 00.0 00.0 00.0 00.0
000 000 000 000 000 000
000 000 000 000 000 000
0.00 0.00 0.00 0.00 0.00 0.00
.mummu _mumm. _mqmm. _wqmm: 0.00. 0.00

Hm.o wmé mN.o om.o

000.0 0.0 0.0 03.0
00 00 00 00

0.0 0.0 0.0 0.0

0 m .0

00.0 00.0 00.0. 00.0
0mm 0mm mpm mum
mmm 000 000 000
0.00 0.00 0.00 0.00
00.00 .. .50 0.00 - 0000

m0

. 000.0

0.00 0.00 _mummu_.umnm. 0.0

 

 

A .0 000 000 . 002.00000002

A .H may :Fzéomonvaz

A .H wé 00000000000200 H.308
A ..H m5 :mwbS 00903003
A00000v 000000000

mm

A .0 000 0000000000

A .-00 10.000 000 00

000000000000

A . Dov 00000000000089 .0303
Toov 0.00—0.00.0323. .004
0.0.0.3 m 0.5% 09300003

A0000V 0 00000000000 00000

 

0000: 060.0 0000:

omma .3 0.008004 .. 020.30 8300 com 003 Mm 000.000.033.330 100.0200 EH .30me

B- 10

mound “Humane“: £9353 0.9% £50.55 .33me .m ..2 Soy.“ dawn

HH.0 mH.0 mm.0 0H.0 0H.0 «0.0 40.0 00.0 00.0 m~.0 m~.0 00.0
0.0 0.0 0.0 0.0 0.0 00.0 00.0 00.0 0.0 0.0 0.0 ma.0
.aa ma 04H 0m 0m 000 mg: 00 H0 H0 m: 0HH
H.m 0.0 m.0 0.0 0.0a 0.0 m.” 0.0 0.0H H.0H 0.HH 0.0
c m a a o o o a m m m. o
ma.~ mH.~ ma.” 04.0 ma.~ 0m.” mm.~ 00.0 m0.0 m0.~ 00.» mp.“
mam 400 000 400 000 000 400 400 000 000 000 000
mma m0; H00 000 fiam omm 000 mum 00m «mm 00m pmm
m.~ m.» m.0H 0.0a 0.0a m.mH m.ma 0.0H 0.0a m.~a 0.0a m.mH
NH.H -.zoq «.ma . swam
00
Ham.m
gfimflawflflaflflflflumflflgflfll m...

 

N

3.33 damaged: «mcwmcwq £05m Qaawoaooo .m .3 80.8. wanna

 

A :H may moz + «02.:mmonaaz
A ..H wev amzéwwonpaz

A .H m5 agonamonm H.309

A ...n mev cmmbno 09363.3
$0.23 $003.39

:9

A 2H may hufiaaawxaq

A ..08 wmano 0-3” 03

hnflpaponccoo

A . oov 0.33.3253 .333

A . oov 23.3359 .34

A. 99.3 m 0.5.: mmhwnoma

Apmmmv m mafia pnmfimm mm0.00

 

9:8: 55 and:

omma .om nonempamm a Hobwm .5“..qu com 2.3 no mofimfihmpoaudzo Hdodnmno new 3".me

00.3.3 :mgowz £50ch pawm namgsm 92.3003 .m .2 E95 3.09m

00.3.3 damage“: £030qu 0000556 Hmoawoaomu .m .3 08km dawn:

B-ll

 

40.0 00.0 00.0 0H.0 0H.0 mfi.0 0H.0 4H.0 mH.0 mfi.0 m~.0 ma.0 A 1H may 002 + «02.00000000
m0.0 00.0 0H.0 0.0 0.0 00.0 mm.0 00.0 0.0 0.0 0.0 0H.0 A 1H 000 002.0000000z
«H 0H 00H 00 H0 mma mmfl H0 00 0: mm mHH A 1H 000 0000000000 00000
m.0 0.0 m.4 H.0 0.0 0.0 0.0 0.0 0.0 H.0H m.0H 0.0 A :H 000 0000x0 000H00000
- A000000 000000000

00.0 m0.0 m0.0 m0.0 m0.0 0m.» 00.0 00.0 m0.0 00.0 mm.0 m0.0 , mm
000 000 000 000 000 000 000 0mm 000 0mm 0mm 0mm A 1H may 00000H00H<
m2 03 0mm 00m fim 00m 00m fim 00m 4% 3m 0am ..00 .050 0.3 5

. 000000000000
m.ma 0.0a 0.4a m.0a m.4H 0.0a 0.0a 0.mH 0.0a m.mH 0.ma 0.0a A.0ov 00000000000 0000:
No.0 .. 33 0.3 .. swam A.oov 0.3.5.8989 HQ

om A.m..w.ov m 3H: 09850.3
00.0 A0000V m 0002 000000 00000

H

543 min mJH .15 momm 06H m.m.n 06H 00mH nooa wow mo: 5.902 50.0...“ 9.0.52

 

 

0m0H .00 000000000 0 00000 00000 000 000 0o 000000000000000 H0002000 000 00000000

APPENDIX C

C-l

mpdm

m .0

0A 7th $5

0 0

p P

.Nmma umsws< mafipsb 00>Hx Adamo 00m 0:» no coauosv00m pcmsww
3390 89G A003”: :3 0050me

0A

A

OH x <5 03:: namfimfiaogm 809: 00.0ch
3th ES 395..“ 9:00.39: mo 0830509050

 

 

00‘

 

 

.‘O‘.

\‘O|Q‘ol
'.|-‘ol
‘0."

pm? .mauo 0.0593. I 000098 3.900.096
Nmma .NH 92803 I 0093an H0355

A03 8

I;

 

 

‘ I
‘0 '

.‘o'olo'o‘

j

 

..H 05 000003: 00000005 00000 .. .ii

A ..H 03 0009008000 00500000 H0000. .. ..-:

 

 

 

/
r

'
§

 

 

. 00m

 

 

 

 

A .mec «.5 OH x 45 3.23 meEmwaofEQ SEE 09056
925 .Nmma pmsmga mfihfi hofim .9600 vmm 23 no 20305ng pcmsmamofinna v5 waged 3333. no comahmasoo

 

H3330 sou.“ 33H: :3 3ng
m 4 m N M m :
A I u w I < _I— - _l— O
\---- .
"'--""--""I” .
\“\\ I! ”
\\\ I —
\\ i -
\\ o
\‘ o c
s\\\ a ..
IIII .. . A. 03
\\\\ a .
\\\ - u
_ u
.. .
.
. .
fl “ If
.
_ n
. .
u .
. .
. .
. .
., . : cow
. .
. .
. .
.. u
. .
M .
’~ 5
$3 .m spam-awn 3 0m pmamaé I @3098. «pang-mafia
wmma .ON pndmné I mom-madam Hmofismgo
. on
A L“ m5 mgonamoza 3:0me H309 I III-I o

 

 

C-3

A 753 «-5 anH 8 <3 and? pcoSMHao-rhzm Sues 30:3
Pam .wmmH avenue-awn ”EH-:6 995m gmo 6mm 93 so manage-Hm pcmEMH .0

£93 98 mHo>mH .3355: .«o couHHmQEoo
HHampao scum AmoHHx HQ 3:335

 

 

 

H o
m #— - M W - u C C C 4 E o
\s s -’\'Q'.|lo’! \.\ A
o‘o‘o\9 'I'lo"/ .‘h'fi'l’
I‘Io'g‘c‘.‘ .--. ,s’\ a A
v o‘.‘.|n|o \\\‘\‘I- ..‘l"' ’ 'M
A .\\\\ III! .I m.
“I “ . [III / w“
I||I I a.-
|||\ II (n
‘\‘| I .
\\\“ ’0 u L
o . OOH.
: o .
. .
. .
. .
— .
a .
.. ..
_ u 9
. .
.v . .
. .
. .
n .
. n
u -
. .
.. n . 8m
. .
.r a u
. .
. .
.. n
4 $3 .nmIMH Amnsmpaom I @3098 «pm-33am
wmmH .MH $92393 I non-338 HmoHsmno
30H K ..H was among-H: ogmuofi Hmpoa I I.I.I
A ..H m3 ogosmnona chomch Hmpoa I 3.3- L com
1

 

 

 

«H-

.A .136 ..Ec OH N 53 mpg 305.3353 :3: 30:06

nnwm .pmmH nopouoo w:H::v yo>Hm havoc we: as» :o coa¢ozvonm p:memeo¢hnm ucm nHmpoH pcoHsps: mo :omHumnsoo
HHprso :95 $032 :3 3:3.an

m a . n N H

1L0

 

 

- c 4 1 . :EECO

‘
\‘\ I
\ I
\

~
_
.
_
_

wmmH .OH :mnopoo I cm nogampmmm I camomxo uprpmnsm
$2 .m “Bo-+8 - ”mm-nis- H3230

A ..H 93 gonamoafl Um>HomnHU Hdaon. I IIIII

 

 

C-S

A 1mg “.5 .3” x «.3 mad; ”Ememwmophnm name Song
953 .umma .Hmnopoo mafigc .825 .868 Umm 23 no coapofivopm pamfimwmophgg 98 ImHm>wH pcowafiuc mo comemQEou
H.335 50.8 “no.3”: c3 00:3qu

m a I m N

 

 

 

. I 4 I w I - I I n.
c : a c c a o
‘I|'III-I'III"II'|‘|I"|IIII ....... lull. IIIII I'll .‘||l|\
Va .. .\
Ivoluc'ollolloI-oluo' a I. .'o\
''''' 0-0‘O|\-o'0'0'0 0’. ‘ ’l a“
’0 9 ’c/ .-
I. z
I. \ a. .-
I.’ s. 96—»
0’. o . 8H
I.I .\
~...\
A
I 8m
umma 3.938252 op Hm nwnopoo I ummoakm ganpmnsw 4
$3 “an honopoo I 3thQO H3205
A .3” K .H may ammonpwd ownwmhofi Hanan. I I.I.I
A ...n m5 3&0:ngan cobaommwc H.309 I IIII m
. oo

 

 

C-6

 

 

.A .-hmu $5 go.” x 53 mpg ammemwmogg 59: 3056
2mm .53.. 38832 mafia 33mm .830 gm map so 53030.3 “Beam...” egg and magma pagans: no nomaumaéoo
H3330 60.5 “mod”: :3 35335
m 4 m m H o
4 1 ‘IE 4 I E 1 q E E 1 j- o
IIIIIIIvIu-l'l-"l'I'nl"||I|-II‘.I-II" \II'".
......I' “
--.l'lll —
I _ .
,, ..
.I.’.I.I. .I/ III» \
I.Io ’ .
I I I . x
./.,.l I .\
.p .\ . 8H
. .\..I
_, _
fl _
._ _
, ._ I
I\
A com
nmma «HNINI .HmQSoboz I 60838 upwaumobm .
R2” .3” umnsmboz I 39338 dwodnmno
Auoa N .H was cmmeficdfiqmmaocw H.309 I III #
A ..H m3 mgozamona ngommflu H4309 I IIII I 00m

 

 

 

 

hmma «OH nopaoomn I om pmnegoz I @3098 upmnpmnam
wmma «m nmnamomn I 39323 $0.255

$3 an .H was names? 398989” ona I

A ..H m3 mfiofimofi 35°38 H38 I 2...:

 

.A .men ~26 “med N 43 n32: pcméfiaophna cums 30:3

98m .Nmma $8.803 wafihfi 35m .3..an «Ex .33 no 5326th pcmemw 81an «Em 396.” €339: Ho comaamQEoo
H3320 Boa.“ A33: :3 @28me o
a
m w c m M r.- » {mu Ed . JIII'J o
"."
. II|I|||IIII"AIIII-II"-I-'|'I'\-‘|‘--‘-"” “
I . .
¢ .
I o u
. .
. .
. .
. .
. .
. .
. . . OOH
n O Q
. .
.
o _
{k
35'. .\.
'0'0'0'0|0'0|0|\'0'0'\l9' lfilOIQ \I‘D\
I .
.I. _
[.1 _
.Io 0). s
I.’.\I\ If“
_ cow

 

“Sm

A 15 $5 «OH M iv
mafia: pnmewamophzm name muocmn mumm .hmma umpemoon mcwudv ho>wm “memo nwm map so soaponcoam acmamwaophrm
HHmMpso echm.Ammez Gav monwpnfia

m a m m a o

 

 

I I ”flu, 4 I rt, I, “unllllnullnuIIIJI1luu1,o

18H

mmma .5 humaqu op wmma «kw hmpfimuon I vmmogxo upmupmnam

 

8m

 

.A 13 .-...6 OH R <5 manna; gcmfiwfiaofmna 838 3.0va

9:5 .33 Sand» mcwhzu .8pr Eco com 23 no 532693 pcmcafimophna v5 naobma 95.3.3: no 483.8930

358 EC 33%: :3 8538

 

 

 

 

m 4 m N H o
1 4 in « I [I1 4 C q HuIIIUIflJ o
' ‘-"““¢II\“‘ """""’ “
"‘--"'l'-'|“"‘ I D
I .
( A
J OOH
.\
o'o'n'o' -\
o‘.‘ ~'.'.IIQII.'.II.II. IIIIII I. \
o\ Iolo’ .
\.\ ..I.I./ .\
o‘c‘ lulclol \\ '0‘ A
.\.\ .1 \.
.\ .(.
\
A 00m
;
mmma .wmIfl “$355 I @8098 32395
mmma 3H 5.35th I 3thQO Hmugono
Auoa N ...n m5 nowoafin 35305" H309 I ii
A ...n was Eggnog nobaommflu H38. I III: I 00m

 

C-lO

A .186 V5 CA N 43 3g pcmfiflnofinna :mme macaw
2mm .umma 339mm mcfihfi 35m 888 vmm 93 no 5302qu pnmaflmogm and magma p523: .«o «53.39.50
H3320 29C Amoduz :3 09.832—
Lm a m a 0

Jo.

 

 

 

4H? J J“ 1 : ii '30

|
I
I
I
I
I

I
-..u”--“
A

)

4/

OOH

.-.-.l

-‘-o

‘.
‘.
\

(

00m

mmma «ma humanmh I mu 555w I @3093 mpwnpmnzm
mmma «w Egnoh I own? Havana

93 H .H was ammonia 35305. 309 I

 

A .H 9.3 agonamofi umpaommflu H33. I
oom

 

 

C-ll

A 153 15 OH x 53 mpg pcméwwnuophna game 3023
2mm .mmma £0.83 wcwhfi 35m .880 vmm 93 no "8.32693 pcmewfimsIEQ 98 3954 9:335: no comaaameoo
.3330 son.“ “mad“: SC 3:339 .
m 4 m N Hg 0

 

 

L1... 'III‘IIIO"'I- I II I -.-II".III!I" Illllllllll"--ll \v lCl— —

 

 

I.
I.
O
I\
'.

\.....

I.
I.
'.
l.
’.
I~
l.-.'.'.'ol.|o'n'.‘-Il.lcllol\

mmmfl .OmIN. scams I @0398 wumupmnsw
wmma Jun :93: I nowhflwnm Hmodnmno

Ago.” N ..H may some»? 35305” H309 I Iii
A .H m3 mgosmmona 62503.3 H.309 I ...:

A

 

 

OOH

OON

oom

C-l2

2mm .33 no.3: 9356 .833 gwo vmm 23 no 20.33605“ ”Ema—w.“

m

A .103 «.5 mod K 43 mpg newsmamofinnm cams 30:3
0

H333 saw 33? a: magma
a , n N H

‘0

En v5. 393..” £33.32 mo conflmeOu

 

 

4

4 1 AW n

..l "|"||||"'|'l‘ r|'|-"' ' |""'|-II'|"--. VJ w..-----"'-'-'k
\ .

IL

 

 

 

o
‘0 '\'\ l ¢
‘0‘. 'c

 

 

Clutr

lo'.'o|o'o|olc'o’.

 

 

 

 

 

 

 

32 .m 3.54 .. cm 33: u conga 38925
32” .3 53 u 393:3 3026.6

:04“ x ..H m5 some»? oguuonfi H309 I Eli

. A ..H m3 mahoganona 350mm? H30“. I 3......

 

 

\I: an

 

 

\- ~-—-—----‘

I

“"1
!
!

 

‘\

 

l_'

 

.8“

 

Tom

 

C-l3

A .53 16

OH x «dv «was; pcmEMfimopth cams mpOCmn

 

mama .mmma awnm< wcahsv uo>flm hmvwo nmm mag co cowooscoua psoemwmophna and nam>oa pcofinpac no soughwgeoo
Hawmpso 80am Ammafiz.:wv ooampnfla .
m I 4 I m N H o
J AW fl 4 I.
r — GIII:IF
0

 

 

 

 

IIIIIIII'IIIIIII

 

 

II- 'I.."Il"'|l"'l'I-"l

mmma .NHIM Hang< I umuogxm wumupmnbm
wmma «ma Hwagq I mwmhamcm Hmofiswno

 

'-'-'-..-".""

|I
‘o‘o‘clls
.‘0

 

Auoa K :H wsv ammonpfi: afiawmaomfl Hmpoa I I;I;I

A 2H mmv mahonamong ampaommflc Hmpoe I I...-

Q‘.

 

at
I

 

I
’0--.-.

.
u
.
o
c
c
c
c
.
o
.
4
I
z. r.
1..

IR

 

r.

 

 

r.

0.0 '0'.

 

 

 

OOH

com

com

A 13 «.5 OH on <5 and? #553335” :38 3056
8.43 .33 3.5.4. wfihfi pofim goo 6mm 23 co cowaosvona £85“ 3193 and magma 302.5: Ho ”Swanson
H.335 30.5 Amman”: 2.3 35me
o

C-lh

' "-"|-"'--"

.-.‘C'. -.-.

\'~' '
l I

OOH

00w

awn .mNIE 3.34 .. 3893 «5535
wmma «mm 3.34 I mmmmgcm 39.828

Auoa x ..._.. may cmwafid 0.23393” H309 I 1!!

A 1H mzv mahonmmonm ©0>HOMMHU Hmvos I IIIIO 00m

 

c-lS

 

A . A ...hmn «.5 mod“ x <3 mug acmemamoffm 52: 30:3
”Hum .mmma Huang mafia 35m .8qu Ex 0

on» no £33695 95$“ fang 9am magma 333% mo comHAmQEoo
H3350 50AM Amwduz 2.3 85.939
m a m w a - o
G 1 N u d o

 

‘|AI‘_-‘—
r'-"||l"l.'l|ll

....rl ""’IIII|IIII II!
‘-‘.-".'...‘-
‘1" ‘ 'I I-‘

r.
'11-
’

L

T.|l.|o'.'o'.'olo'ol 0L

'olo'o '0 Io'c‘o'Q|o 's'ol.-. :l.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

4 00m

 

mmma .m has”- mm Hagg< I omnogxo «paupmaum
mmma .0m Hfiaqq I momhamqm Hmofl5¢no
Auoa x ....” 95 names? 0.28985 309 I Iii

A .H m3 agonaaonm 330me dmpoe I IIII com

I;

C-16

 

A .anc .15 H x 45 3% 952389139 508 30:3
2mm .33 mm: mafia .823 .8va Ex «5 co 2030:ng £83.“ ofiEa can. magma ...—Swans: Ho :omfiBQEoo
H.335 Sop.“ 30.3”: 93 00503:

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

m w H o
L A 'l’. m I“ 'I‘W b‘b ‘ I o
0'0'0'0 4'10“" l0|0|o“olo|b'0'o'|-0 v"h|’ul A ,o'oIoIOl 0“.“ nuqh
\1 1.1:? N
sl' |I \\\\ / A
11 ill!" ----‘LTJt|""-nl'-".."l.lll III I, _I ~ —
T I; w i
x ,. m,
I; a m.
I u
I ’1 “Ar lukn r
. 8H
if . J
m
r n
F
F r t r m
_ , 4
a
a ._
.. "
fl . L 8w
1 n u
.. w
_ u
. _
. .
. .
w . d
T mmma .mHnm hm: .. @3093 «paupnnsm
3m." .3 am: .. $955 €3.35 .
A ..S x L 9: comps? ogmuofi H38. . i...
A ..H was agonmmonm ©230me H.309 I 10::

con

 

C-17

.A _. OH x <5 and? pcoemfimoga 5mg 38%
2mm .mmmH mm: mghfi .823“ .8an com 05 no couposoogm Epcwpcmfimoifi 98 3.254. naming“ mo nomahngoo

admmpso Scam 33%: 2.3 vacuumed

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

m a m N H o
‘ C '1. 7! £0 “W I x ‘N’ o
o‘t'o|oLIlvo'o-olvo[o'o'o'o o
1 i. .. 3...... m... fix
I Isl. \o “o I. I -------: -
lclt'o’olrnlro“\u -- I““" c’ "
-|'I|'|II-.|III.IIIIIIIIIIIU . u
T I, """" 'iI|IUUII'.L ‘1 I.‘ .“‘ m L
I" u
l a n
’ a
fi E (x f
+ c r 2 §
. i
L i 08
,
r
P 33 .24.; am: .. 3398 33933 +
wmma «ma ads I mmmhawnd awofismno

 

A .H m3 maogamonm c0503? H309 .. null

 

dam

C-18

 

 

Ema
o poswoha new

m 95% Ao>am Hdvmo cam mna no a H

. hm: ca

mhmm mmma

 

L ‘|"' I

 

can
mpfica pcmswaaophha cmwwawwmsoo
.A wzwu .Eu kowhmmdwww mHo>oH vamfluvsc mo

m
HAMHpso saum Ammafiz adv cocwa fin

 

 

 

 

 

 

o
.. A
..- ,. 1..
IIL-‘II""" “ mom- EOOH
. mu”
.
. .
n u
m u
. u 4
. .
. .
. .
.
.
. .
n .
fl . t 8m
. n
. .
.
. a
. n
o -
Rx i
can
I uumonxo_maanpm
mmfimmmmum Mm .. @835 fiofimno
w 08 ' o'c'
N H msv ammonpfic oacmwnoqa amp
Auoa 1

 

com
0 I. 'III B
A .H mav mphonamona vm>aommav Hun a
.I

 

[Hi
H

 

 

C-l9

mpmm

m

.A Thaw #26 %OH N «dv mpflns pcosmwmoemnm cams prCmv
.wmma mash mcfihsw hm>wm Macmo vmm man no cowposthm pqmewa ophnm cam mHm>mH vcmflhpsn no confinwasoo
Hawmpso 39C Ammdwz :3 028me
: m m H o

mmma quI4 82. I @3098 dpmfipmpsm
wmma aw." mash. I uwmhdmcm H3230

:0." H ...n was comgpfi: 03935” .....309 II.I.I

A .H m5 agonmnoza vozommflu 309 I IIII

 

C-20

.A .anu «-5 OH H 45 mpg: acmemeofEm 5.9: 305%
98m .33 05:. waist .823 .3ch gm 9.3 no 20.30395 pcmfimHmofEm EB mHm>wH pcmHHflE Ho somflwano

HHdepo Eon.“ AmmHH: 2.3 39325

 

 

 

 

 

 

 

 

 

 

m a m a H o
l «I! L If - I [4 ET 0
‘o . .|.|.l. lg:
.‘o't'g|o-o‘. ‘0 u ‘.
ko'o'o|o|o-.'.‘6'. | .‘~‘~l.'o'.' I"I‘- -..-“---- L1"Ol. “*Ilil i
““|"‘|\I|n I ”—0 _u A
l t|l|llOI'IlIII|"I.|IFII|I-I.'I|‘I.|“|‘ ——I 0“
... a ..
2 .n
I a.— — .
n. .. A SH
ark“
— .
H .
. .
n .
. .
. .- u A
a
.
I Has-
I
J oom
mmmH HomImH 92:. I @3098 «$335 4
mmmH .2” 93A. I mmmAHdcm HmoHEmno
A N0H K ..H was «Swap? OHQmeocH H309 I ....I.I
Q A ..H m3 maofimonm @050me Haven. IIIIII

 

 

 

 

com

 

 

E6 .5 n2 x is
mpflqd acmemflaophzm cams mpocmv wamm .mmma mash mcwhnt hw>wm hmvmo mom mnp co coaa.o:v0hmwpcmewfioophnm

HHSS 58.“ 33H: :3 Snags
m 4 m m H o

|F ¢ E 1 4d d R! :flir-Eo

 

 

C-21
T

 

Hp>mH amp“: cw omdn poem av
mmmH «CA a op om ocaw ummOQXm apahpmnfim

 

00m

C-22

. .A 7>wv $26 ea N <<v mews: pcmEMflmophcm cmms wpocmn
mhwm mmma hash mcfihsv Ambfim hmvmo vmm . .hopmsa cam wHw>¢H pcmwnps: mo zomwamqsco

 

 

 

 

 

 

 

 

 

 

 

m a H338 29¢ 3on: :3 8533
m
.allcllc sllullsllg'.lu.lloll.'.l!oll~l{ICI-Iisll.-\'.llo|lol \“nlufllnv .
‘c'o"r.’.il nl.|unl.nl.'c|\l L III-"II"('_L W. [I

. ‘-------- / . a +
C \\\...\\\.. /. fl _—
‘\1\ I. ..
III-“II"IIII‘II‘II“ 0. o—

.Ilu‘ll ’o m. *Lkv 8.”

...-J-
383-5
C..-
--

fa

33 .312“ 3.3. n 3398 32925 . Ra
mMmH «mm hHSfi I mmmhdmcm HmowEmso

A «OH x .H may comps? ogmaofi H309 n Iii

 

A ..H m3 maofimofi 3:0me H309 u ...---

‘1
1“
lll‘l

 

 

mhwm .mmmH 92%: E
4 $5.28 .33“ . In <4 H5 5w
. . .m gm m A - an I a I
. o m mmofi co cofloHfiVa paumfiwame “a v If. pew .3319?” came m
Hm azo Boga A mmHHz c3 mozmpanE PB mHoboH 30.29:: Ho comHeHmemv
. . o

 

C-23

 

 

 

 

m
I m. , , m N
O O'Olo‘olo|o'o 0| 11 II
o'o'o'olo'o'o‘o‘o‘o|o‘1o' 0‘0‘0‘o'o|opn<mlg . I I I I m
. o o‘ o I I O
E I. A
I. A
L ,. T:
a .. (I
I|“Il‘\I-"“I--' o v I
||'I'I'II‘|‘\I|I|““||.|||| I" h A

III‘.-|II-I'|'I \“|III."‘ do 0‘
'I' 0‘ a
... .u
e. _“
... _n

c. a L 03
fl. ..n
.t.
u u
.. m

. '0‘. A

4 Dow

a
mmmH m pmsmgmwmofmm .HHHHH. I 95098 mamhpwob
a H H .32. .. «mafia HSEBM

o K
Au H ..H m5 ammonia UHSmeHoHHH H38. I

A ..H m3 mgofimona cm>HommHv H309 I
. 00m

 

 

.A as «-5 "a x s:
mpflcd aswawwaophzm cums mvocmw mhwm .mmma pmsmq< mnwgdu hm>fim udvmo Umm wnp co soapostopa pcwewwnoahnm

Hammuso EOLH Amoawz may moamamwa

 

0-224

 

 

 

 

 

 

 

 

 

 

a m w H o

4 1 J« «H 4, —I— ~I— 1 Fl— 0

ll .

f

L r
.02

A
ASN

mmma .mano pndm:<.cmmogxm upmhpmnsm

 

oom

 

 

APPENDIX D

Nutrient levels of the Red Cedar River at Station -O.b

Date
July 5, 1957
27
August 6
12
20
September 13
October 3
31
November 19
December 5
January 114, 1958
February 6
March 11
26
April 8
' 15
22
30
May 12
15
27
June 2
10
19
July 1
5
7
9
23
31

Phosphorus (28_I“ )

 

Dissolved Sestonic Total AmmofiIh

28

51
h6

28
29
2h

31 .

22
18

13
16

13

15
23
37

35
32

26
53
h?

109
117
88
106
113
79
88

107

000 OOOOOOOOPPOOOOOOOOOOO
OHU OOOOOOOO l-‘l-‘HO 0000000

.00
CO

1.86

D-l

)
otal

Nitro en (mg I“
2+

1 .86

0.28
0.31

0.61
0.72
1.3h
1.0h
0.99
1.59
1.06

D-2

Nutrient levels of the Red Cedar River at Station -O.2

 

 

_T__Phospgprus (pg I“ ) Nitrogen (m I“ )
Date Dissolved Sestonic Total Ammonia N02 + 803 Total
July 5, 1957 230 0.0 8.12 8.12
27 33 88 77 0.0
August 6 26 52 78 0.0
12 bl Sh 95 0.0 0.21 0.21
20 h? 89 96 0.0
September 13 bl 30 71 0.0 0.hl 0.Ll
October 3 hh 21 an 0.11
31 3h 8 L2 0.22 0.50 0.72
Nevember 19 27 28 55 0.10 0.96 1.06
December 5 2h 28 52 0.11 1.h1 1.52
January 1h, 1958 19 33 52 0.1h 1.27 1.hl
February 6 23 33 56 0.16 0.72 0.88
March 11 19 20 39 0.0h 1.72 1.76
26 18 21 39 0.0 1.00 1.00
April 8 15 bl 56 0.06 1.09 1.15
15 13 29 82 0.0 0.86 0.86
22 18 66 80 0.0 0.59 0.59
30 1h 88 62 0.0 0.23 0.23
May 12 17 37 Sh 0.0 0.11 0.11
15 17 37 5h 0.0
27 22 so 72 0.0 0.0
June 2 29 69 98 0.29
10
19 30 L2 72 0.18 0.28 0.82
July 1 25 63 88 0.0 0.11 0.11
5 hb 86 130 2.70
7 ho 88 88 1.73
9 28 62 90 1.80
23 27 h? 78 0.0 0.29 0.29
31 37 60 97 0.0 0.06 0.06

Nutrient levels of the Red Cedar River at Station *0.1

Date
Jfl)’ 59 1957
27
August 6
12
20
September 13
October 3
31
Nevember 19
December 5
January 1h, 1958
February 6
Mbrch 11
26
April 8
15
22
30
May 12
15
27
June 2
10
19
July 1
S
7
9
23
31

Phosphorus (

'Dissolved Sestonlc

99
208
2h8
280

85

98

133

126
59
88

150
93

252
82
280

162
222

1&8
156

63
103
59
90
88
117
69
22
62
65
90
83
St
110
63
15h
12h
288
26
152

h8
158

I” )

Nitrogen (mg 1|."| )

D-3

otal 'Immonia N02 +NO3 Total

230
115
202
267
338
328
202
167

97
195
109
216
102
1&2
260
156
230
272
500
108
392

210
380

220

0.0

0.31
0.37
0.70
0.87
0.51
0.32
0.53
0.32
0.60
0.28
0.65
0.19
0.31
0.57
0.25
0.37
0.50
0.69
0.32
0.78

h.12

0.18

0.21
0.05

8.12

Nutrient levels of the Red Cedar River at Station 40.3

Date

October 3, 1957

31

November 19

December 5

February 6

March 11

26

April 8

15

22

30

May 12

15

27

June 2

10

19

July 1

5

7

9

23

31

Phosph

 

Dissolved

78
68
88
82
35
86
26
31
25
30
35
88
87
53
57

86
85

83
51

orus ( 13 ) Nitro en (mg I“ )

Sestonic Total Ammonia §5E+ N03 :Tbtif
82 116 0.27
33 97 0.17 0.86 0.63
23 67 0.16 0.58 0.70
36 78 0.20 1.51 1.71'
83 78 0.22 1.39 1.61
39 85 0.23 0.96 1.16
26 52 0.07 2.00 2.07
27 58 0.05 1.13 1.18
89 78 0.18 1.13 1.27
33 63 0.06 0.86 0.92
72 107 0.10 0.75 0.85
60 108 0.08 0.36 0.80
57 188 0.15 0.13 0.28
58 111 0.16 0.12 0.28
61 118 0.05 0.16 0.31
50 96 0.25 0.28 0.89
78 119 0.0 0.22 0.22
60 103 0.83 0.28 0.67
60 111 0.35 0.03 0.38

0-8

 

Nutrient levels of the Red Cedar River at Station +0.6

 

 

Phosphorus (pggl'I ) Nitrogen (mg 1"I )
Date Diesélved SestonicA:T3t51’ Ammonia N02 # N03 STEtal
July 5, 1957
29 86 57 103 0.0
August 6 36 60 96 0.0
12 86 56 102 0.0 0.18 0.18
20 58 59 113 0.0
September 13 78 87 121 0.11 0.31 0.82
October 3 50 35 85 0.18
31 38 28 62 0.16 0.97 1.13
November 19
December 5 31 29 60 0.15 1.67 1.82
January 18, 1958 30 26 56 0.19 1.27 1.86
February 6 33 22 65 0.17 0.82 0.99
March 11
26 22 22 88 0.0 1.00 1.00
April 8 26 38 68 0.10 1.20 1.30
15 17 33 50 0.0 0.89 0.89
22 15 .70 85 0.0 0.61 0.61
30 23 88 71 0.0 0.86 0.86
May 12 88 60 188 0.08 0.18 0.22
15 26 85 71 0.0 0.12 0.12
27 81 57 98 0.0 0.08 0.08
June 2
10
19 82 52 98 0.16 0.10 0.26
July 3 37 75 112 0.0 0.12 0.12
7
9
23 82 58 100 0.06 0.25 0.31
31 52 66 118 0.0 0.06 0.06

May

June

Nutrient levels of the Red Cedar River at Station +1.3

Date

12, 1958
15
27
2
10
19

Phosphorus_£pg I'| )

Dissolved Sestonic

26
30
88

88

86
88
63

67

O

72
78
111

111

Nitrogen (Efi 1"| )
onia 2 9‘ 3 013

0.00
000

0.15

0.10
0.12
0.11

0.11

0.10
0.12
0.11

0.26

Nutrient levels of the Red Cedar River at Station +1.8

Date

July 5: 1957
27
August 6
12
20
September 13
October 3
31
Nevember 19
December 5
January 18, 1958
February 6
March 11
26
April 8
15
22
30
May 12
15
27
June 2
10
19
July 1
5
7
9
23

31

qummus(mif')
‘DEESOIved Sestonicfi_Tbt§1 Ammonia N 2 +

320
77
86

101

113
97

115
52
62
60
57
65
82

122

ee
00

e
5’

090090000
HHOOO
O

O
\O

O .00
00000080

0.00 00000000
0
OHM
O\\O

0?
08

e
O

2.13

0.18
0.31

0.68
0.90
1.33
1.07
0.96
1.92
1.00
1.28
0.59
0.51
0.80
0.10
0.08
0.06

0.26
0.31
0.17

0.25
0.08

3

D-7

Nitro en ( I“ )
0 80 Total

2.13

0.18
0.31

0.78
0.99
1.85
1.21
1.12
1.96
1.00
1.30
0.59
0.51
0.80
0.10
0.08
0.06
0.28
0.55
0.37
0.17
2.80
1.60
1.80
0.31
0.08

F”

D-8

Nutrient levels of the Red Cedar River at Station *3.7

Phosphorus Lug;1“ ) Nitrogen (mg IJ )

 

 

Date Dissolved Sestonic ‘Tibal Ammonia N02 + N03 T3721
July 5, 1957
27 81 35 76 0.0
August 6 78 28 102
12 108 23 127 0.0 0.25 0.25
20 118 26 180 0.0
September 13 9O 19 109 0.0 0.51 0.51
October 3 73 28 97 0.06
31 86 18 60 0.11 0.65 0.76
November 19 32 27 59 0.09 0.60 0.69
December 5 39 26 65 0.12 1.33 1.85
January 18, 1958 38 26 68 0.16 1.00 1.16
February 6 8O 28 68 0.20 1.27 1.87
March 11 22 26 88 0.08 1.62 1.66
26 18 25 83 0.0 0.67 0.67
April 8 17 39 56 0.0 1.28 1.28
15 17 33 50 0.0 0.68 0.68
22 27 89 76 0.0 0.82 0.82
30 28 38 62 0.0 0.80 0.80
may 12 37 38 75 0.0 0.10 0.10
15 88 37 85 0.16 0.16
27 68 38 102 0.0 0.08- 0.08
June 2
10
19 65 28 93 0.11 0.30 0.81
July 3 91 88 135 0.0 0.28 0.28
7
9
23 93 33 126 0.0 0.39 0.39
31 88 37 121 0.0 0.13 0.13

Nutrient levels of the Red Cedar River at Station +8.9

Date

January
February

March

April

May

June

July

18, 1958
6

11
26
8
15
22
30
12
15
27
2
10
19 .
1
5
7
9
23
31

Phosphorus (ug_I‘ )

 

Tfigsolved Sestonic Total

35
35

19
2O
20
27
26
37
50
71

65
97

100
85

30
38

20

80
39

0.17
0.20

0
00000000

000 00900000
H\O

e
OHM

CO
e

1.50
2.08

0.89
1.08
0.57
0.37
0.80
0.03
0.16
0.07

0.35
0.33
0.27

D-9

Nitro en (m I“ )
Ammonia 562 v 563 Total

1.67
2.88

0.89
1.08
0.57
0.37
0.80
0.03
0.16
0.07

0.68
0.88
0.27

0.35
0.13

APPENDIX E

Physical and chemical characteristics of the Red Cedar River

July 5, 1957

27
August 6

12

20
September 13
October 3

31

November 19
December 5

January 18, 1958

February 6
March 11
26
April 8
22

as determined at Station 03.7.

Water Tgmperature
( C.)

20.0

22.0

20.0
21.0
21.0
20.0
11.5
6.5
5.0
0.5
0.0

0.0

8.0
7.0
18.5

4 C171")

Conductivity
(x 10“ ohms

638

588
519
562
860
866
580
621
573
610
515
528

510

Y

Alkalinit
(mg I )

192
296
286
276
266

‘ 260

270
270
228
272
276
288
220
288
226
266

pH

7.9
7.9
7.9
8.1
8.2
8.1
8.0
7.8
7.7
8.0
7.9
8.0
8.1
8.8
8.1

8.3

“833188”

52
68
38
17
26

w

19
12

15
22
23
15

25
22

M USE ONLY
p “881 USE 8381

 

 

[fl-1. ‘D

7m

£59.23); ‘3', a! :-

x.‘ cm ”XWCM‘ -‘ ‘

... <vh-‘

 

 

.«e.
\.
O
1‘.t ...)“ n
f V I. U ' ‘ u. 4n
6 .. x r . .
b , ..7. ., a .
a). I .Q!‘ at“! e .. ’1.
h (I-
“'o ,0 0"
.. 7.‘
D
‘ ‘» lv
d e. .
I ' a
0 .. p
7 “A . . a?
p ‘03.,
‘9’. 12‘.
' :5? e
I 2.)...
s... 7'0.
7.... a ..
a,
vi . we
0 And 101.
‘. It a
Q‘
a . .. , 4..
I .I \J ’ x Q L ' ’65! .\ .4 l.. ’e L .‘ 1
1 .. .5730?!
... .
d
..n
1a..
..s.‘
a . .e.
. a ... \rm .1
e s.
. e ..a ‘ .
.‘g \

nICHIan STATE UNIV. LIB'RRRIES
VIHIHWIW“WWWWWIHNUHIIMHIWI
31293006585636