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ABSTRACT

EFFECTS OF A MUNICIPAL DISCHARGE
ON A MICHIGAN STREAM

BY

Richard L. Mikula

A survey was conducted on a southern Michigan
stream during the summer and fall of 1973 to determine the
changes in water quality caused by sewage effluent.

Changes in water quality were detected by measuring changes
in heavy metal concentrations in bottom sediments, primary

productivity, phytoplankton communities, fish communities,

and macroinvertebrate communities.

Macroinvertebrate and fish communities were dras-
tically altered for over 2.5 miles below the wastewater
treatment plant (WWTP). Fish were virtually eliminated
and the macroinvertebrate community consisted of large
numbers of individuals that were pollution tolerant. Along
this portion of stream, turbid and septic smelling water
covered a black anaerobic substrate of sludge.

PhytOplankton communities were relatively uniform
throughout the stream and were dominated by diatoms tolerant

to organic enrichment. Diatoms which are less favored by

‘w‘dE

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.nu

hue

Richard L. Mikula

sewage than green and blue—green algae comprised slightly
higher concentrations of the algal community above the
wastewater treatment plant than below.

Mean primary production, based on chlorophyll a
production, experienced a 4-fold increase below the WWTP
for at least four miles. Primary production remained
approximately 2.5 times larger than the control station for
the remaining portion of the stream.

Concentrations of all heavy metals (arsenic, cad-
mium, chromium, copper, mercury, nickel, zinc) in the
organic sediment increased below the WWTP and remained
higher than the levels above the WWTP for the remaining

portion of the creek.

EFFECTS OF A MUNICIPAL DISCHARGE

ON A MICHIGAN STREAM

BY

Richard L. Mikula

A THESIS

Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Fisheries and Wildlife

1975

ACKNOWLEDGMENTS

I would like to express my appreciation to Dr.
Eugene W. Roelofs for his guidance and advice during this
study and to Dr. C. D. McNabb and Dr. C. R. Humphreys for
reviewing this manuscript.

I also wish to thank the Michigan Water Resources
Commission for making it possible for me to conduct this
study and for providing the staff to analyze the sediment
and chlorophyll 3 samples. Special thanks go to Ronald
Willson, Dr. E. Evans, George Jackson, and Richard Lundgren,
biologists in the Stream Survey Section, for their help in
algal and macroinvertebrate identifications and for,
reviewing this manuscript.

Lastly, I would like to express my sincere
appreciation to my friend, Dr. Tracy S. Carter, who pro—
vided considerable help and encouragement throughout this

study.

ii

TABLE OF CONTENTS

Page
LIST OF TABLES O O O O O O O O O O O O O V

LIST OF FIGURES. . . . . . . . . . . . . viii

INTRODUCTION. . . . . . . . . . . . . . 1
BACKGROUND . . . . . . . .' . . . . . . 2
METHODS O O O O O O O 0 O O O O O O O 4
Phytoplankton. . . . . . . . . . . . . 4
Periphyton. . . . . . . . . . . . . . 8
Sediments . . . . . . . . . . . . . . 12
Fish Populations. . . . . . . . . . . . 13
Macroinvertebrates . . . . . . . . . . . 15
RESULTS . . . . . . . . . . . . . . . 22
Habitat. O O O O O O O O O I O O O O 22
Phytoplankton. . . . . . . . . . . . . 22
Periphyton. . . . . . . . . . . . . . 26
Sediments . . . . . . . . . . . . . . 27
Fish Populations. . . . . . . . . . . . 29
Macroinvertebrates . . . . . . . . . . . 29
DISCUSSION 0 O O O O O O O O O O 0 O O 39
LITERATURE CITED . . . . . . . . . . . . 46

iii

Page

APPENDICES

Appendix
A. PHYTOPLANKTON DATA . . . . . . . . . . 49
B. PERIPHYTON DATA . . . . . . . . . . . 52
C. ORGANIC SEDIMENT CONCENTRATION . . . . . . 54
D. FISH AND MACROINVERTEBRATE DATA. . . . . . 56

iv

Table

1.

Al.

A2.

A3.

B1.

B2.

B3.

LIST OF TABLES

Station location and habitat descriptions for
sampling sites on Sycamore, Willow, and Mud
Creeks and Vevay Drain in the vicinity of
Mason and Lansing, Michigan, July 25 to
November 8, 1973 . . . . . . . . .

Phytoplankton densities in Sycamore Creek and
Vevay Drain in the vicinity of Mason and
Lansing, Ingham County, Michigan, July 2,
1973. Data expressed in organisms/ml .

PhytOplankton densities in Sycamore Creek,
Willow Creek, Mud Creek, and Vevay Drain in
the vicinity of Mason and Lansing, Ingham
County, Michigan, September 5, 1973. Data
expressed in organisms/ml . . . . . .

Phytoplankton densities in Sycamore Creek,
Willow Creek, Mud Creek, and Vevay Drain in
the vicinity of Mason and Lansing, Ingham
County, Michigan, November 8, 1973. Data
expressed in organisms/ml . . . . . .

Periphyton standing crop collected every two
weeks in Sycamore Creek, Ingham County,
Michigan, July 25-Septemb3r 5, 1973. All
values expressed as ug/cm of chlorophyll a

Two-way analysis of variance for'adjusted
periphyton primary production from Sycamore
Creek, July 25-Sept. 5, 1973 . . . . .

Comparisons of differences between mean
standing crOp from Sycamore Creek, July 25-
September 5, 1973. Data expressed in
ug/cm2 of chlorophyll a. . . . . . .

Page

23

49

50

51

52

53

53

Table Paqo

C1. Heavy metal concentrations found in bottom
sediments of Sycamore Creek and Vevay Drain,
Ingham County, Michigan, July 25, 1973.
All values expressed in mg/kg, dry weight . . 54

C2. Concentrations of chlorinated hydrocarbon
pesticides, polychlorinated biphenyls,
phthalates, and oil found in the bottom
sediments of Sycamore Creek and Vevay Drain,
Ingham Co., Michigan, July 25, 1973. All
values expressed in mg/kg. pH values
expressed in standard units. . . . . . . 55

D1. Summary of fish data collected by electro-
fishing at selected stations from Sycamore
Creek, Ingham Co., Michigan, Sept. 7, 1973. . 56

DZ. Aquatic macroinvertebrates taken in quali-
tative collections from Sycamore Creek,
Willow Creek, Mud Creek, and Vevay Drain,
Ingham Co., Michigan, September 4-5, 1973 . . 57

D3. Estimated macroinvertebrate numbers per
square meter in Sycamore, Willow, and Mud
Creeks, Ingham Co., Michigan, based upon
colonization of Hester-Dendy artificial
substrate samplers, July 25-September 5,
1973 . . . . . . . . . . . . . . 59

D4. Summary of macroinvertebrate data from
Hester-Dendy artificial substrate samplers
placed in Sycamore, Willow, and Mud
Creeks, Ingham Co., Michigan, July 25-
September 5, 1973 . . . . . . . . . . 60

D5. Number of species collected on circular
Hester-Dendy artificial substrates in
Sycamore, Willow, and Mud Creeks in the
vicinity of Mason and Lansing, Ingham
County, Michigan, July 25-September 5,
1973 . . . . . . . . . . . . . . 61

D6. Species diversity (5) of macroinvertebrates
collected on circular Hester-Dendy
artificial substrates in Sycamore, Willow,
and Mud Creeks in the vicinity of Mason
and Lansing, Ingham County, Michigan,
July 25-September 5, 1973 . . . . . . . 61

vi

Table Page

D7. Comparisons of differences between mean number
of macroinvertebrate species collected on
Hester-Dendy artificial substrates in
Sycamore, Willow, and Mud Creeks in the
vicinity of Mason and Lansing, Ingham County,
Michigan, July 25-September 5, 1973 . . . . 62

D8. Comparisons of differences between species
diversities of macroinvertebrates collected
on Hester-Dendy artificial substrates in
Sycamore, Willow, and Mud Creeks in the
vicinity of Mason and Lansing, Ingham County,
Michigan, July 25-September 5, 1973 . . . . 62

vii

LIST OF FIGURES

Figure Page

1. Selected station locations in Sycamore, Mud,
and Willow Creeks and Vevay Drain in the
vicinity of Mason and Lansing, Ingham
County, Michigan, July 2-November 8, 1973 . . 5

2. Artificial substrate utilized to sample
periphytic algae at selected locations in
Sycamore Creek in the vicinity of Mason
and Lansing, Ingham County, Michigan,
July 25-September 5, 1973 . . . . . . . 10

3. Circular Hester-Dendy artificial substrate
sampler colonized by macroinvertebrates
in Sycamore, Mud, and Willow Creeks,
vicinity of Mason and Lansing, Ingham
County, Michigan. July 25-September 5,
1973 . . . . . . . . . . . . . . l7

4. Total number of species, proportion of each
tolerance group, and biotic index deter-
mined from qualitative samples collected
in Sycamore Creek, Willow Creek, Mud
Creek, and Vevay Drain in the vicinity
of Mason and Lansing, Ingham County,
Michigan, Sept. 4-5, 1973 . . . . . . . 30

5. Estimated number of organisms/m2, proportion
of each tolerant group, and the number of
species collected on circular Hester-
Dendy artificial substrate samplers
placed in Sycamore, Willow, and Mud
Creeks in the vicinity of Mason and
Lansing, Ingham County, Michigan,
July 25-September 5, 1973 . . . . . . . 34

viii

Figure Page
6. Species diversity, equitability, and biotic
index values determined from colonization
of organisms on circular Hester-Dendy
artificial substrate samplers placed in
Sycamore, Willow, and Mud Creeks in the
vicinity of Mason and Lansing, Ingham
County, Michigan, July 25-September 5,
1973 . . . . . . . . . . . . . . 36

ix

INTRODUCTION

A biological and sediment chemistry survey was
conducted on Sycamore Creek and three of its tributaries
(Willow Creek, Mud Creek, and Vevay Drain) in Ingham
County, Michigan, between July 2 and November 8, 1973.

This survey was conducted to assess the water quality and
habitat of the stream from above Mason to Lansing, Michigan,
a distance of approximately twelve miles. There are three
known discharges within the study area: Wyeth Laboratory
Company in north Mason which discharges cooling water into
a storm drain, Mason Wastewater Treatment Plant (WWTP), and

Dart Container Company which discharges into Vevay Drain.

BAC KGROUND

Sycamore Creek originates approximately 4.5 milesf
south of Mason and empties into the Red Cedar River in
Lansing, Michigan. Total length of Sycamore Creek is
approximately 16.5 miles with a drainage basin of 111
square miles. The 7-day, 10—year low flow at the mouth of
Sycamore Creek is 3.5 cfs (Knutilla, 1968). Flows are
normally much larger than this, exceeding 6.4 cfs 95 per-
cent of the time and 14 cfs 70 percent of the time.

The Michigan Water Resources Commission (MWRC) has
conducted two previous surveys in this same area (Basch
et a1., 1971; Riley, 1972). In March and April, 1971, an
i§_§i£g bioassay was conducted by Basch et a1. (1971) to
determine the effects on fish of the chlorination operation
of the Mason WWTP. Results of this survey showed that
chlorinated compounds discharged by the Mason WWTP were

toxic to rainbow trout (Salmo gairdneri) for at least

 

 

0.8 miles downstream and to fathead minnows (Pimephales

promelas) for at least 250 yards downstream from the WWTP.

 

Riley (1972) conducted a continuous-flow bioassay

to determine the toxic effects of the Dart Container

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Company's effluent to fathead minnows. Bioassay results
showed fathead minnows subjected to Dart's effluent could
not survive in concentrations greater than 50 percent
effluent. The effluent contained oil, high concentrations
of total solids (highest level was 3720 mg/l) and pH values

ranging from 8.8 to 12.0.

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METHODS

To assess habitat and water quality in Sycamore
Creek, nine sampling sites were selected (Figure 1). In
addition, one site was selected on both Willow and Mud
Creeks, and two sites on Vevay Drain to determine the
effects of these tributaries on Sycamore Creek. On Vevay
Drain sites were selected both above and below the Dart
Container Company discharge to determine any water quality
degradation in the drain and ultimately in Sycamore Creek.
Phytoplankton, periphyton, sediment, fish pOpulations, and
macroinvertebrate communities were used to assess the water

quality in this survey.

Phytgplankton

 

Plankton algae in standing water are a reflection
of the toxic or nutrient status of the water which they
inhabit. This is not true of river algae however, because
algae present in river water samples may have come from a
considerable distance upstream. Plankton in a stream
develops only where the current is reduced, such as along
stream margins or in pool areas (Patrick, 1948). Benthic

and periphytic algae are often dislodged from their natural

Figure 1. Selected station locations in Sycamore, Mud,
and Willow Creeks and Vevay Drain in the
vicinity of Mason and Lansing, Ingham County,
Michigan. July 2-November 8, 1973.

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habitats by river current scouring and appear as "plankton."
Therefore, planktonic algae collected at various stations
reflect upstream water quality. However, a change in water
quality will be reflected by a change in planktonic algae
between stations.

One-liter surface water grab samples were taken for
plankton analysis at all stations (Figure 1) on July 2
(except stations MC-l and WC-l); September 6, and
November 8, 1973. Samples were preserved with 75 ml of
6:3:1 water—ethyl alcohol-formalin preservative and
returned to the MWRC biological laboratory in Lansing,
Michigan, for qualitative and quantitative algal analyses.
Samples were allowed to settle for 48 hours and then they
were drawn down so only the bottom 100 ml remained. This
100 ml concentrate was thoroughly mixed and subsampled.
Samples were analyzed utilizing Sedgewick Rafter strip
counts. Permanent slides were made for diatom determina-
tions.

After the algae samples were identified and tabu-
lated, a pollution index value (Palmer, 1969) was determined
for each station utilizing algae that occurred with a

frequency of 50 or more cells per m1.

P.I. = X]. + X2 + X3 +0....O...+ X20

where P.I. = the pollution index value and X through X

1

equals the numerical values assigned to the twenty most

20

pollution tolerant genera (Palmer, 1969). An index value

of 20 or more is evidence of high organic pollution, while
a value of 15 to 19 is considered to be probable evidence
of high organic pollution (Palmer, 1969). Lower figures

indicate organic pollution is not high.

Periphyton

 

Periphyton is the total assemblage of plants
growing on the surfaces of objects submerged in water
(Young, 1945). Since periphyton, along with rooted aquatic
plants, are considered the most important primary producers
in a stream system, changes in periphyton production
reflect changes in water quality.

Primary production can be calculated by determining
the amount of chlorOphyll a present in the periphyton
growing on the substrate (Hynes, 1970). ChlorOphyll a
is the photosynthetic pigment in fresh water plants and is
directly related to the amount of green plant material
present. By comparing the amount of chlorophyll 3 produced
in a given time period per unit area, the production rates
in a stream can be determined.

Major factors influencing the amount of production
are nutrients, light, turbidity, velocity, temperature, and
toxicants. By selecting areas along the stream with simi-
lar physical conditions, major changes in productivity can
be attributed to changes in nutrient levels within the

water and/or presence or absence of toxic substances.

Artificial substrate samplers (Figure 2) used to
measure primary production in this survey were constructed
by bending a 22-inch length of 3/8-inch threaded rod at a
right angle, making one arm about 15 inches long and the
other about 7 inches long. Four blocks (3 x 3 x 4 inches)
of high density styrofoam were connected to the rod, three
on the long arm and one on the short arm. A one-pound lead
weight was attached to the rod below the single block of
styrofoam. The three blocks on the long side caused the
sampler to float while the lead weight pulled the single
block down below the water surface, thus providing substrate
for periphyton.

After selecting appropriate locations, steel posts
were driven into the stream bed for sampler attachment.
Six-foot lengths of 1/4-inch chain were used for attachment
allowing the samplers to move up and down with water level
fluctuation.

Periphyton growing on the submerged block was
collected three times at 14-day intervals as recommended
by King and Ball (1966). Collections were made August 8,
August 22, and September 5, 1973. Thin slices of the
3 x 3 inch ends of the submerged block were cut with a
knife and placed into a 250-ml black bottle with 50 ml of
90 percent acetone. The black bottle was required to
prevent the breakdown and decay of the chlorophyll a while
enroute to the laboratory for analysis. Acetone dissolved

the styrofoam and served as a chlorophyll extract. The

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acetone-chlorophyll extracts were sonified and spectro-
photometrically analyzed for chlorophyll a (Standard
Methods, 1971) by the Michigan Water Resources Commission
pesticide laboratory in Lansing, Michigan.

Two-way analysis of variance was performed on the
data to determine if significant differences between sta—
tions and sampling dates had occurred. Tukey's w-procedure,
described by Steel and Torrie (1960), was used to determine

which stations differed.

Sediments

 

Organic sediments are a major repository of heavy
metals, chlorinated hydrocarbons, and oil in the aquatic
environment. These materials, if discharged into water
quickly become attached to organic particles and settle out
in slow moving or still water. Organic sediments therefore
serve as a useful monitor of previous discharges of these
substances.

Samples of the organic deposits were taken at each
station (Figure 1) except stations MC-l and WC-l, on
July 25, 1973. Samples were carefully collected by taking
the upper layer of organic sediment by hand and placing it
in a sterile Whirl-Pak plastic bag. Samples were kept cool
until they were returned to the MWRC wastewater laboratory
in Lansing, where they were frozen until analyzed. A
portion of each sample was analyzed by atomic absorption

techniques for the following heavy metals: arsenic, cadmium,

l3

chromium, copper, lead, mercury, nickel, and zinc. The
remainder of each sample was analyzed for chlorinated
hydrocarbon pesticides, phthalates, polychlorinated
biphenyls (PCB's) and oil by using standard gas chroma-

tography procedures. Sediment pH was also determined.

Fish Populations

On September 7, 1973, resident fish pOpulations
were examined at six selected locations (SC-1, SC-2, SC-S,
SC-6, SC-7, and SC-8) on Sycamore Creek (Figure 1). Fish
collections were made with a backpack electrofishing unit
utilizing a 12—volt battery. Stunned fish were collected
with nets and placed in a bucket until shocking was com-
pleted. Each area of stream, approximately 2000 square
feet, was shocked for 45 minutes. Species identifications
and tabulations were made on site with the exception of a
few species which were labeled, preserved with 150 ml of a
formalin solution (37% formaldehyde, 63% water and
methanol), and returned to the MWRC biology laboratory in
Lansing, Michigan, for identification.

The collected fish were categorized into the
following trophic levels defined by Willson (1972):

Class 1 These fish are nondifferentiating
bottom feeders. They live, grow, and
multiply in clean water areas and will
also thrive in waters of degraded
quality. In low water quality areas
the variety of macroinvertebrates is low

but total numbers are very high. The
straining of food organisms from sludge

l4

deposits is easy and large quantities
of food are available at little expense
in energy.

Group 1: Fish in this group are moderate to large
in size. They suck or rasp off bottom
animals such as midges and aquatic worms
from rocks, sand and bottom ooze, expel-
ling substrate materials and consuming
the animals. Examples of fish in this
group are carp, suckers, sheepshead, and
bullheads.

Class II These fish species are differentiating
predators and are generally associated
with clean water environments. They
live, grow, and multiply best when a
diverse macroinvertebrate food supply
is available.

Group 2: Fish in this group are small in size and
as a result are restricted to feeding on
smaller macroinvertebrates. They are
generally classed as forage species.
Examples of fish in this group are
minnows, darters, shad, shiners, and
chubs.

Group 3: Fish in this group are moderate to large
in size and are capable of utilizing the
entire macroinvertebrate community in
addition to fish in group 2 and fry of
groups 3 and 4. Examples of fish in
this group are crappies, bluegills, sun—
fish, perch, and catfish.

Group 4: Fish in this group are large in size and
utilize fish from groups 1, 2, 3, and 4
as their principal food source. They
may also utilize larger macroinverte-
brates from the benthic community, but
fish comprise the major portion of food
intake. Examples of fish in this group
are pike, bass, bowfin, and burbot.

By categorizing fish into the above discrete trophic
levels it is possible to utilize fish community structures
in evaluating existing water quality conditions (Willson,

1972). A weighed trophic index (T.I.) is used in this

15

evaluation. T.I. = x(l) + x(2) + x(3) + x(4), where x =
the number of fish species in a trophic group, and l, 2, 3,
and 4 = the trophic group numbers. Thus, species most
dependent upon higher water quality conditions and a stable
balanced community are given greater weight than those
capable of withstanding degraded water quality and the
resulting community imbalance. Higher trophic index values
will therefore reflect more stable community structures and
higher water quality conditions with lower trophic index

values reflecting reduced water quality conditions.

Macroinvertebrates

 

Aquatic macroinvertebrates and their community
structures are useful in evaluating water quality. Since
these animals spend their entire life or parts of their
life cycle (eggs, larvae, and/or pupae) in the water, they
can reflect long term water quality conditions to which
they have been exposed.

Generally, a natural unpolluted stream will support
many different species but only a few individuals per
species due to competition for food and living space
(Gaufin and Tarzwell, 1956). In polluted streams many taxa
are eliminated, but those that remain usually occur in high
numbers due to lack of competition.

Qualitative macroinvertebrate samples were taken
at all stations (Figure l) on September 4-5, 1973. Samples

were collected with triangular dip nets and by hand picking

16

from all habitats found at the station site. Time of
collecting was approximately 30 minutes or until no new
species were found with additional sampling. Samples were
placed in quart jars, labeled, preserved with 100 ml of

a formalin solution (37% formaldehyde, 63% water and
methanol), and returned to the MWRC biology laboratory in
Lansing, Michigan.

Quantitative samples were taken with modified
Hester-Dendy artificial substrate samplers at all stations
(Figure 1) except stations V-A and V—B. On July 25, 1973,
two samplers were suspended with wire in the water column,
approximately 6-8 inches from the bottom. These samplers
consisted of a five-inch eyebolt and 8 circular plates of
1/8 inch tempered hardboard with a diameter of 2 3/4 inches
(Figure 3). Small hardboard spacers (3/4" x 3/4" x 1/8")
were positioned so that 1/8, 1/4, and 3/8 inch spaces
existed between the plates. Plates and spacers were held
in place by a washer and hex nut. Each sampler had 0.061
square meters of surface area available for colonization by
macroinvertebrates.

On September 5, 1973, following six weeks of eXpos-
ure, the substrates were carefully removed from the water
by placing a #10 sized can with a U.S. Standard #30 mesh
screened bottom below the sampler and slowly bringing it
up around the sampler. The wire suspending the sampler
was cut and the sampler and material in the bottom of the

can were placed in a quart jar, labeled, and preserved by

Figure 3.

17

Circular Hester-Dendy artificial substrate samp-
ler colonized by macroinvertebrates in Sycamore,
Mud, and Willow Creeks, vicinity of Mason and
Lansing, Ingham County, Michigan, July 25-
September 5, 1973.

19

covering the sampler with 95 percent methyl alcohol. Sam-
plers were returned to the MWRC biology laboratory in
Lansing, disassembled, and the individual sampling com-
ponents scraped with a putty knife into a container with a
U.S. Standard #30 mesh screen bottom. After sieving and
washing, the remaining material was placed in labeled vials
containing 75 percent ethyl alcohol.

Macroinvertebrates from both the qualitative and
quantitative samples were identified and tabulated. Each
identified taxon was assigned a tolerance status suggested
by the United States Environmental Protection Agency
(Anon., 1973).

Tolerance status refers to the animal's relative
ability to withstand and/or respond to adverse environ-
mental conditions. Individual tolerances are generally
derived from an animal's reaction to organic wastes and
attendant oxygen depletion or modification of bottom
deposits.

Tolerance status is generally defined by the
Michigan Water Resources Commission as:

Intolerant--organisms whose growth and development are
dependent upon a narrow range of environmental con—
ditions. They are rarely found in areas of organic
enrichment. They cannot adapt to adverse situations

and are replaced by less sensitive organisms if the
quality of their environment is degraded.

 

Facultative--organisms with the ability to survive over
a wide range of environmental conditions. They possess
"medium" tolerance and often respond positively to
moderate organic enrichment but cannot tolerate severe
environmental stresses.

 

20

Tolerant--organisms that can grow and develOp within a
wide range of environmental conditions. They are often
found in water of poor quality. These species are
generally insensitive to a variety of environmental
stresses.

Benthic communities can be compared at the various
stations by calculating a biotic index (Beck, 1955) which
weights the number of species according to their tolerance.
The biotic index (BI) is calculated by doubling the number
of intolerant species and adding this to the number of
facultative species (BI = 21 + F). Values for the biotic
index normally range between zero and 40, with polluted
streams usually having values less than 10 (Beck, 1955).
Biotic indices were calculated for both the qualitative
and quantitative samples.

In addition to the tolerance status, the diversity
of the animals present in a given benthic community is
significant. Species diversity was determined for the
quantitative samples using Shannon's formula as described
by Patten (1962):

_ s

d = lilni/N log2 ni/N
where "N" is the total number of organisms, "ni" is the
number of individuals per species, and "s" is the number of
species. The value of d ranges from zero to any positive
number. Wilhm (1970) states that d rarely exceeds nine

and is generally between three and four in clean water and

less than one in polluted stream areas.

21

When degradation is at slight to moderate levels,
5 lacks the sensitivity to demonstrate differences. Dif-
ferences can often be detected by calculating equitability
at each station. The equitability formula (Lloyd and

Ghelardi, 1964) used was:

where s = number of taxa in the sample, and s' = the number
of species expected from a community that conforms to the

MacArthur's model in Lloyd and Ghelardi (1964). Streams

unaffected by oxygen demanding wastes generally have e
values ranging from 0.6 to 0.8 while streams with even
slight levels of degradation have "e" values generally

below 0.5 (Anon., 1973).

RESULTS

Habitat
Station habitat descriptions, locations, and obser—
vations are shown in Table 1. From these data the immediate
deleterious effects of the Mason WWTP are apparent. Sand \
and gravel, the usual substrate occurring in Sycamore
Creek, waskcoveredwith black anaerobic sludge_for a dis-
tance of greater than 2.5 miles below the WWTP. Clear

water above the WWTP became turbid andwhad a septic smell

for a distance greater than 2.5 miles below the WWTP.

Phytoplankton

 

Data from the three sampling runs are presented in
Appendix A, Tables 1, 2, and 3. July algal abundances
ranged from 680 (SC-5) to l4l9/ml (SC-6). Organisms per
ml decreased from 1206 above the Mason WWTP to 904 immed—
iately below the WWTP at station SC-3. Decreases in algal
counts continued downstream to station SC-6, where the
highest counts occurred (1419/m1). Diatoms comprised over
93 percent of the algal communities at each station except
station SC-9 where diatoms made up 87 percent of the

community. Green (Chlorophyta) and blue-green (Cyanophyta)

22

23

usmpcsnm couzzmfiuom
ucoEepom as can woum3 so

mxcmn Bowman
co unsunm can mmmum

unmeflpmm pcm

acmqaou
uuuo 3o~wn

 

Hwo «unwao no: noun: “mmuznmouome ofiuusvm oz .Hm>mum .pcmm m.H o.e o.H pnom puma mu>
ucmpcsnm cu3oum owuzcm hammaoo
ufiumd “ponusumwp was mxcmn Emouum uocfinucoo
ucmEHowm cwnz oomwmamu so unsuzn can mwuum ucoEwpmm can undo o>onu
Hwo nummao mc3 noun: “mmuznaouomfi oHumsqa oz .Hm>uum .ocwm o.H o.v o.H boom xomnmmom ¢n>
mopsHm Amazz
mono oHumwm Us: mmpsHm mxcmn Emmuum no mo muwmomop coma: konn
can umum3 «uoHoo c“ mnsunm can mmuum “mnmxa was: nus: nudge h.ov
ouazsnnmazmum mma nouns can mommnum vacuumem oouo>oo team o.m o.mH m.o puom Haozom vnum
pm>uwmno who: mzoGCAE mapsan
“Loco owumwm Um: oopsam nxcmn Eowuum so no muwmommv @933 count
can umum3 «uoHoo a“ mmouu can .mnsunn .mmmum mos: "Ho>ouo 30H0n
mufinzunmflzuum mu: umumz “mommmum ucmmuwem mmummm can pawn unaudm m.~ o.~H o.an.o noon ooH mnum
pm>u0mno @933
mum3 zwmpwsoumoumnuv mxcnn Emmuum :0 «won» noun: o>ono
mumxozu can n3occaE can .mnsunm .mnmuw xoou new oucuuuco
“unmao was nouns «nouznmouooa caumsvo oz .Ho>oum .Ucom m.a o.oa o.~no.H zuouosoo .m «tum
mxcmn Eomuuu co Ho>mum hm .m.D
madman can ammum Hana no mucsosd Eoum
unmau ma: noun: “mouznmouumfi vaumsqw oz Hanan :ufi3 pcmm o.H o.n o.Hno.o Emmuumma Hi0:
pm>uomno
mu03 Ammpflcaumzov axcmn Emmuum co vamp sown:
msoccfle usouofisc Issac woman can mnsunm no nusom
«Human mm: nouns “mommmum ucomumso omuMQm pawn new aw>muu o.H o.m o.H pmom mafia Alum
unowuu>uumno codumuwom> oumuumndm «any sumoo Auuv nupwz oom\um noduuooq umhfidz
Honuo omnuo>< muoum>m 3on noduoum newumum

 

.mnma .m umnEm>oz 0» mm zHSH .cwmflnoaz .mcwmcnq can comm: no zuficflow> onu aw
cacao >m>0> can mxmouu on: use .3oHHw3 .ouosmozm co mmuam mcflHQEMm you anewumwuomop Havana; can cofluuooH Godumumnu.a mqmda

241

 

unaccsna
zusoum owuznmfiumm .ucmE
lupmm mnu cw Hwo “vwnusu

xeun
Edmuuu so nndunm pan

 

usuawpmu can

Am833
noun: 30H0n

nmdda o.HHv

 

zflunouau uw3 nouns manna “uwmmnum unmoumsm .xoou .pcum o.v o.o~ w.o .cm «mom .uz atom
guano Amazz
mnu macaw unwed GOunz sodon
pm>uwmno mum: mxcun Edwuum wnu :0 Boum loom «axoou can modes m.mv
macaque "human was umuoz mmmum “uwmmmum ucmmuwem .Hm>mum .pcmm m.N o.om m.H Uuom added alum
nmmpm
mson ucme Amezz count
mxcmn co upon no muumoamp soaon mouse
ownusu 30um momuu can unaunm Hausa “axoou can m.hv pnoz
zaunmuam mm3 noun: umwuzcmouoms Ouumsqm oz .Ho>num .Ucwm o.m o.m~ o.~uo.H omua Guam snow
Amazz
Emcu ou uooo vuumou uxcun Edmuum :0 3oum mxoou 30w sown: 30Hon
uzmflau a on: uawm can mumuo can unsunm «mwuznm a pan .uawn amuse o.vv
uwun3 “human um3 umuo3 nouomE vuuusqn oz .Ho>mum .Ucmm o.v o.m~ o.auo.o boom uaoz olom
Eoouun
wzu no ummuo ca ucmmoum
ov>ummno mum) Amaocfiowuuo Esuuusummzv Odom
m3occua “unmao an: nouns mmouuumumz mmoH new pcmw m.H o.oa o.~um.o mafiaaasm duo:
was:
ucwfiuoon onu uxcmn wmpsau Gown: Spawn
cw duo no noouuu «uoHoo Edmuum co mmoum :uu3 pmuo>oo awaas m.~.
cw uuuszlnmuzoum uouuz “mommmuo ucomuoem Hm>wum paw wcmw o.v o.om m.o coon Magnum mnum
ncOwuu>uwmno :Owumuwmm> oumuumusm zuuv sumac Auuv sup“: omu\uw cOwuuooq nonadz
uozuo wmmuw>¢ momum>¢ 30am noduoum cOuuoum

 

.Umscfiucounl.a mamas

25

algae combined, comprised less than 5.0 percent of the algal
communities at all stations except station SC-9, where
these groups comprised 10 percent of the community.
Flagellates comprised less than 3.0 percent of the algal
communities. Palmer's pollution index ranged from 6 to 9
(Appendix A, Table 1).

September algal abundances ranged from 961 (SC-l)
to 2792/m1 (SC-6) in the main stream. Diatoms comprised
over 90 percent of the algae except for stations SC-4
(70%) and MC-l (84%). Green and blue-green algae combined,
comprised less than 6.0 percent of all samples. Flagellates
comprised less than 4.0 percent of the algal communities
except at stations SC-4 (25%) and MC-l (15%). Palmer's
index values ranged from 6 (SC-5) to 11 (SC-4) in the
mainstream. Vevay Drain had an index value of 3 below Dart
Container Company.

November algal abundances ranged from 95 (SC-l) to
432/ml (SC-4). Diatoms comprised over 90 percent of the
algal community at all stations, except stations SC-3 (33%),
SC-4 (79%), and SC-8 (88%). Green and blue-green algae
comprised less than 5.0 percent of the algal communities,
except at stations SC-2 (9%) and SC-8 (8%). Flagellates
comprised less than 6.0 percent of the algal communities
except at stations SC-3 (62%) and SC—4 (16%). Palmer's
pollution index ranged from 3 to 6.

Nitzchia and Navicula, ranked by Palmer (1969) as

the sixth and seventh genera most tolerant to pollution,

26

were dominant at all stations on all sampling dates.
Navicula was generally more abundant than Nitzchia at most
stations and sampling dates except in September when
Nitzchia was more abundant than Navicula at stations SC-4
and SC-S.

Euglena, the most tolerant genus to organic pol-
lution (Palmer, 1969) occurred in low numbers (0-36 per ml)
at all stations and sampling dates except at station SC-4
where there were 260 per ml in September. In November,
Euglena was virtually absent. It was observed at stations
SC-4 and V-B in the qualitative scan, but was only found

at station SC-8 in the quantitative count (1 per ml).

Periphyton

 

Periphyton data are presented in Appendix B,
Table l. The over-all mean standing crops of periphyton
at stations 1, 5, 6, 7, and 8 were 1.70, 6.75, 6.72, 3.90,
and 4.61 ug/cm2 of chlorophyll a, respectively. Periphyton
standing crop was approximately 4 times greater at stations
SC-S and SC-6 below the Mason WWTP than above Mason at
station SC-l.

Two-way analysis of variance showed a significant
(95%) difference of periphyton standing crop between the
stations (Appendix B, Table 2). Data analysis using
Tukey's w-procedure showed periphyton standing crops at

stations SC-S and SC-6, below the Mason WWTP, were

27

significantly (90%) greater than the periphyton standing

crOp at station SC-l above Mason (Appendix B, Table 3).

Sediments

 

Heavy metal concentrations found in organic sedi-
ments from Sycamore Creek and Vevay Drain are given in
Appendix C, Table 1. Mean concentrations of all metals
except cadmium were higher than Michigan background levels
(Hesse and Evans, 1972). Cadmium was below the limits of
detectability (0.2 mg/kg) at all stations except stations
SC-3 and SC-9 which had respective wet weight values of
0.8 and 0.6 mg/kg.

Mean arsenic (3.3 mg/kg) and mercury (3.2 mg/kg)
concentrations were 1.7 and 11.0 times greater than the
respective mean background levels plus two standard
derivations. Other metal concentrations exceeded back-
ground levels by factors ranging between the above extremes.

Chromium, c0pper, lead, mercury, nickel, and zinc
concentrations increased between stations SC-l and SC-2.
Lead was the only metal which increased substantially
between these stations (29.7 to 102.8 mg/kg). All metal
concentrations increased between station SC-2 above the
Mason WWTP and station SC-3 below the WWTP. The extremes
were arsenic and nickel which increased 1.9 times (2.1 to
4.1 mg/kg and 19.3 to 36.0 mg/kg, respectively) and Copper
which increased 5.5 times (27.8 to 153.2 mg/kg). Concen-

trations of all metals, with the exception of copper,

28

which remained about the same, decreased between stations
SC-3 and SC-4, but were still higher than levels found at
stations SC-l and SC-2 above the Mason WWTP. Metal concen-
trations generally declined in a downstream direction to
Pine Tree Road (SC-7). Below this station all metal con—
centrations increased. Substantial increases in lead

(50.4 to 360.4 mg/kg) and zinc (137.6 to 303.3 mg/kg)
occurred between stations SC-8 and SC-9.

With the exception of cadmium which was below the
limit of detectability, metal concentrations in Vevay Drain
below Dart Container Company were approximately double
those found above Dart Container Company. These levels
were substantially lower than the levels found in Sycamore
Creek (SC-4) above the confluence of the two streams.

No unusually high concentrations of chlorinated
hydrocarbon pesticides, polychlorinated biphenyls (PCB's)
and phthalates (DEHP) were found in any of the organic
sediment samples (Appendix C, Table 2). Sixty-two percent
of the concentrations of the above substances were below
the limits of detectability. The phthalate concentration
(5.2 ppm) below the Mason WWTP at station SC-3 was high
for streams but was average for values detected below
Michigan WWTPs (Hesse, 1973).

The highest oil concentration (841 ppm) was found
below Mason at station SC-2 above the Mason WWTP. Oil
concentrations increased from 412 to 648 ppm below Dart

Container Company in Vevay Drain.

29

Sediment pH in Sycamore Creek and Vevay Drain

ranged from 6.8 to 7.4 (Appendix C, Table 2).

Fish Populations

Data from electofishing are presented in Appendix D,
Table 1. Fish populations were adversely effected below the
Mason WWTP discharge. The number of species decreased
from 10 at station SC-2 immediately above the WWTP to l at
station SC-S, 2.5 miles below the WWTP. Fish numbers
corresponding to the above stations also decreased from
78 to 6, respectively. Fish populations partially
recovered at station SC-6 where 6 species and 18 individuals
were collected. Station SC-7 had the highest quality popu-
lation of fish with 58 individuals and 13 species,
including 2 species in group 4. Station SC-7 had the
highest trOphic index value (28) while station SC-5, 2.5
miles below the Mason WWTP had the lowest trophic index

value (2).

Macroinvertebrates
Qualitative sample data are presented in Appendix D,

Table 2. As illustrated in Figure 4, macroinvertebrates
decreased in number of species from 38, with 4 intolerant
forms, above the Mason WWTP (SC-2) to 9 species, with no
intolerant forms, immediately below the WWTP (SC-3).

Biotic indices for these stations decreased from 36 to 6,
respectively. Species numbers increased at station SC-4 to
23, with 5 tolerants and no intolerants. Two tolerant taxa

(Oligochaeta, Chironomus sp) made up over 85 percent of the

30

.mnma
.muv .umom .cmmflzofiz .zucsou EmnmcH .mcflmcmq paw :Ommz mo zuflcflofl>

wnu cfl gamma zm>m> cam .xwmuu U52 .xwmuu 30HHH3 .xwwuu mHOEmozm
CH Umuomaaoo mmHmEmm 0>Humufiamsq Eouu pmcwauwump xmpcfl oeuofln cam
.msoum mocmumaou 30mm mo cofiuuomoum .mmfiommm mo Hones: Hmuoa

.v musmflm

31

w

To!

.00 F200
.5... <0
n mu) 4..)

xmoz. 02.03

m>....<.530<u

._.Z (uni—O...

hfiKNJOfiZ.

c

a... 3 3
20m<2

n

N

703

 

mzorrdhm
O

O.

0.

ON

mm

on

mm

cc

0?

on

SBIDBdS :IO UBSWflN

32

individuals collected at station SC-4. Seventeen species
with one intolerant, were collected at station SC-S, 2.5
miles below the WWTP.

Number of species and biotic index increased sub-
stantially between stations SC-5 and SC-6 (Figure 4).
Thirty-six species were collected at station SC-6, resulting
in a biotic index of 36. Forty-two species were collected
in Mud Creek (MC-l) which enters Sycamore Creek between
stations SC-S and SC-6. Stations SC-7 and SC-8 continued
to show improvement with station SC-8 being the highest
quality water and habitat found (47 species, BI=46).
Species dropped substantially between stations SC-8 and
SC-9. Thirty-one species were collected at station SC-9,
giving a biotic index of 29.

Quantitative data from Hester-Dendy artificial
substrates are given in Appendix D, Table 3 and summarized
in Appendix D, Table 4. Total number of macroinvertebrate
species decreased from 23 at station SC-2 above the WWTP
to 5 species at stations SC-3 and SC-4 below the WWTP.
Species numbers increased to 13 at station SC-5 and to 28
at SC-6. The highest number of species were collected in
the tributaries at stations WC-l (29 species) and MC-l
(30 species).

Mean numbers of macroinvertebrate species were
significantly (95%) lower at stations SC-3 and SC-4
immediately below the Mason WWTP (Appendix D, Table 7).

Mean number of macroinvertebrate species was 16.5 at

33

station SC-2 immediately above the WWTP and decreased to
4.5 and 4.0 at stations SC-3 and SC-4 respectively
(Appendix D, Table 5). Midges (Chironomidae) comprised

100 percent of the individuals at station SC-3 (Appendix D,

Table 4). One midge species (Chironomus sp) comprised over

 

95 percent of the individuals at station SC-4.

As illustrated in Figure 5, estimated number of
macroinvertebrates per square meter increased substantially
between stations SC-l (983 individuals) and station SC-4
(4713 individuals). Estimated number of macroinvertebrates
per square meter decreased substantially at station SC-S
(2058 individuals) but the number of species increased
from 5 to 13 between stations SC-4 and SC-S, respectively.
Number of macroinvertebrates increased in a downstream
direction below station SC-S and SC-8 where the highest
numbers were collected. At station SC-8, estimated number
per square meter was 7036. Of the total number of indi-
viduals at station SC-8, 40 percent were sensitive mayflies
(Ephemeroptera) and 48 percent were caddisflies (Trichop-
tera). Estimated number of macroinvertebrates per square
meter substantially decreased at station SC-9 to 1386 with
all mayflies and nearly all caddisflies eliminated
(Appendix D, Table 4). Midges comprised 96.5 percent of the
organisms (Appendix D, Table 4).

Species diversity values were lowest at stations
SC-3, SC-4, and SC-S below the Mason WWTP (Figure 6).

These stations had respective species diversity values of

Figure 5.

34

Estimated number of organisms/m2, proportion
of each tolerant group, and the number of
species collected on circular Hester-Dendy
artificial substrate samplers placed in
Sycamore, Willow, and Mud Creeks in the
vicinity of Mason and Lansing, Ingham County,
Michigan, July 25—September 5, 1973.

ESTIMATED MACROINVERTEBRATES lm2_

35

 

eooc ’

1000 I INTOLERANT

TOLERANT

5000
[:l FACULTATIVE
4000

NO. OF SPECIES
( ) COLLECTED

)-

3000 -

I000 __ (20)
900 -

:300L
700-
600»-

   

500 -

400 -

 

 

W

 

 

 

 

 

 

 

 

300

(5)

A

2
9'.

W

 

 

 

 

(28)

 

 

 

(20)

 

 

 

 

 

 

A
(II
V

 

 

 

I
STATIONS l WC'I 2
M

I

4

(3%"

I

Y

I

5

RAIN
C NT. CO.)

MC‘I

6

.1

d

1

 

Figure 6.

36

Species diversity, equitability, and biotic
index values determined from colonization of
organisms on circular Hester—Dendy artificial
substrate samplers placed in Sycamore, Willow,
and Mud Creeks in the vicinity of Mason and
Lansing, Ingham County, Michigan, July 25-
September 5, 1973.

37

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 l I— I I I l I F l I ‘1 i5
).
I: I“) r
g 3 "' ..._.. r—( J3
r—1 7"“
5'! I" --
5 I— 2
2 C
8
'6
g " H I I ‘ I
0 . l—l .
I.O ’ 3.0
" 03 03
:- r—i .
Ii
5 0.6 0-6
g Q 0.4
a
m 0, , , . _ , s 7 7 , 7 ,0
35 35
{—1
3° - «I130
F
25 - J25
("—1
f, 20 - —1 _ -20
o
E
2
I-é I5 r- IS
a:
IO - IO
5 - :55
o H F] o
1 l I I 1 I T I 1 I
STATIONS I W0! 2 I 3 4 I 5 MCI 6 7 8 9

MASON VEVAY DRAIN
WWTP (DART CONT. CO)

38

1.4, 0.3, and 1.5 (Appendix D, Table 6) and were signifi-
cantly (95%) lower than all other stations (Appendix D,
Table 8). Station SC-4 was significantly (95%) lower
than stations SC-3 and SC-S. All other species diversity
values ranged from 2.6 to 3.9.

High equitability values (>0.75) were found above
Mason at stations SC-l and WC-l. Equitability values
decreased to 0.45 at station SC-2 immediately above the
Mason WWTP. Stations SC-4 and SC-5 below the Mason WWTP
had equitability values of 0.28, the lowest values found.
Stations MC-l, SC-6, SC-7, and SC-8 had equitability values
ranging from 0.38 to 0.47. Station SC-9 had an equita-
bility value of 0.69, which is a substantial increase from
station SC-8 (0.41).

Biotic index values (Figure 6) were highest in the
tributaries. Willow Creek had a value of 32 and Mud Creek
a value of 34. Station SC-2 immediately above the Mason
WWTP had a biotic index value of 24 while stations SC-3
and SC-4 below the WWTP had values of 4 and 2, respectively.
Station SC-6 below the confluence of Mud Creek had a biotic
index value of 28, due to insect drift from Mud Creek and
an increase in water quality as a result of higher quality
dilution water entering via Mud Creek. Biotic index
values decreased from 21 at station SC-8 to 12 at station

SC-9.

DISCUSSION

Phytoplankton data did not decisively show changes
in water quality in relation to the Mason WWTP. Although
algal densities fluctuated, community structure was
generally uniform throughout and dominated by diatoms
tolerant of organic enrichment. Diatoms, which are less
favored by sewage than green and blue-green algae (Wager
and Schumacher, 1970) comprised slightly higher concen-
trations of the algal community above the Mason WWTP than
below the WWTP in July and September. In November, sub-
stantial decreases in diatoms at stations SC—3 and SC-4
below the WWTP occurred.

Increased primary production below the Mason WWTP
indicated increased nutrients in the stream. For at least
4 miles below the WWTP production was 4 times greater than
above. Below this 4-mile stretch, production was at least
2 times greater than above the WWTP. When there is an
excessive amount of primary production, fish and macro-
invertebrates mortality may occur because of severe
oxygen depletion and excess carbon dioxide (Hite, 1973).

As can be seen in Figure 5, this did not occur in Sycamore

39

40

Creek. The number of macroinvertebrates per unit area
generally increased in a downstream direction to station
SC-8, where the highest numbers occurred.

All metal concentrations increased between station
SC—2 above the Mason WWTP and station SC-3 directly below
the WWTP. Metal concentrations generally declined in a
downstream direction below SC-3 to SC-7, a distance of
7.5 miles. Concentrations at this point were still higher
than those found above Mason at station SC-l. Below sta-
tion SC-7, all metal concentrations increased from unknown
sources.

With the exception of cadmium, mean metal concen-
trations in the organic sediments were higher than Michigan
background levels (Hesse and Evans, 1972). Sediment metal
concentrations above the Mason WWTP were higher than those
found in nonindustrial streams in Illinois and metal con-
centrations below the WWTP were substantially higher than
those found in the Illinois River (Mathis and Cummings,
1973).

Lethal metal concentrations are extremely difficult
to determine because of limited work in this area and
because lethal concentrations vary with pH, temperature,
water hardness, and other chemical parameters (Doudoroff
and Katz, 1953). Synergistic effects with other metals
also vary the lethal levels of metals (Doudoroff, 1952).

Mathis and Cummings (1973) found that metal con-

centrations in the sediment were three to four orders of

41

magnitude greater than concentrations in the water.
Assuming this same relationship for Sycamore Creek, the
nickel concentration at station SC-3 and SC-4 would be
approximately equal to the concentrations found by Garton
(1968) to be detrimental to aquatic macroinvertebrates.
Also, assuming this same relationship, concentrations of
arsenic, cadmium, COpper, and zinc at stations SC-3 and
SC-4 range from one to two orders of magnitude lower than
concentrations permitted in drinking water (McKee and
Wolf, 1963). Chromium and mercury concentrations at
stations SC-3 and SC-4 were equal to those concentrations
permitted in drinking water, while lead was approximately
one order of magnitude higher than permitted (McKee and
Wolf, 1973). When lead concentrations decreased to per-
missible drinking water standards (SC-6, SC-7, and SC-8)
the macroinvertebrate communities improved (Appendix D,
Tables 2 and 3). When the lead concentrations again exceed
the permissible drinking limits at station SC-9, the
macroinvertebrate community declines again. Lead may not
be the major factor causing the macroinvertebrate responses,
but it is worth considering.

Fish data indicated reduced water quality below the
Mason WWTP. Fish were virtually eliminated for 2.5 miles

below the WWTP. Six fish of one species (Umbra limi) were

 

collected at station SC-S. Fish habitat was ideal in the

vicinity of station SC-S, characterized by fallen logs,

42

deep holes, and isolation from human populations, but high
quality fish communities were absent.

Chlorine was probably the primary waste product of
the Mason WWTP which eliminated the fish. Zillich (1972)
indicates that free chlorine concentrations greater than
0.05 mg/l are lethal to many fish species. Basch and
Truchan (1974) found that continuous exposure to chlorine
concentrations greater than 0.02 mg/l could be detrimental
to intolerant warmwater fish. Basch et a1. (1971) found
the average chlorine residual of the Mason WWTP's effluent
to be 2.64 mg/l. They also found chlorine concentrations
of 0.046 and 0.013 mg/l at stations SC-4 and SC-S respec-
tively. No chlorine concentrations were measured during
this survey. However, there are four reasons why chlorine
might have been the primary factor for the absence of fish:
(1) The chlorine concentrations measured by Basch et al.
(1971) were probably diluted by high stream flows while
concentrations during this survey were probably less diluted
because of low stream flows. (2) Fish in warmer water could
be effected by lower chlorine concentrations than fish in
cooler waters (Gordon, 1974) (Basch et a1. conducted survey
in the spring). (3) Prior to the time of fish sampling,
mechanical failures occurred within the WWTP (Marquardt,
1974)1 so possibly more chlorine was added to the effluent

due to the increased volume of raw sewage. (4) Possibly

 

1James Marquardt, Operator of Mason WWTP, Personal
Communication, Feb. 7, 1974.

43

the chlorine concentrations were not lethal to the fish,
but stressed them so they avoided the four mile stretch of
stream below the Mason WWTP.

Other possible factors effecting the fish pOpula-
tion are: (1) Low oxygen concentrations. (2) Concentra-
tions of heavy metals in the water and sediment. (3) Little
food was available to fish that hunt by sight because the

dominant macroinvertebrates were Oligochaetes and Chironomus

 

which burrow into the sediment.

Macroinvertebrate data indicated reduced water
quality below the Mason WWTP. The number of species and
the number of intolerant individuals were greatly reduced
or eliminated for a distance of at least 2.5 miles. Sub-
stantial improvements occurred in the macroinvertebrate
community at station SC-6, a distance of 4 miles below the
WWTP. At this point sludge was no longer present and Mud
Creek's high quality water had diluted the poor quality
water of Sycamore Creek.

Sludge was probably the dominant factor involved in
altering the macroinvertebrate community. Ellis (1936)
states that silt alters aquatic communities through
screening out light, changing heat radiation, and retaining
organic materials and other substances which create unfav-
orable conditions. Gaufin (1958) also found that the
settling of fine solids form a blanket over the stream

bottom, thus reducing the number of available habitats.

44

Factors discussed in eliminating the fish population could
also have been involved in altering the macroinvertebrate
community.

The immediate effects of the macroinvertebrate
community caused by the Mason WWTP were similar to the
effects of other wastewater treatment plants (Mackenthum,
1969; Olive and Dambach, 1973; Gaufin and Tarzwell, 1956).
Mackenthum (1969) found that organic pollutants in the
absence of toxic materials cause dramatic increases in the
densities of Oligochaetes and chironomids. Olive and
Dambach (1973) found that these organisms (Oligochaetes
and chironomids) accounted for over 90 percent of the
invertebrate community. These two groups made up over
90 percent of the invertebrates collected at stations
SC-3 and SC-4 in both the qualitative and quantitative
samples (Appendix D, Tables 2 and 3). Gaufin and Tarzwell
(1956) found that there were usually one-third to one-sixth
as many species below a sewage plant than above and that
these organisms (Hemiptera, Diptera, Coleoptera) usually
have special respiratory modifications that permit them to
survive in poorly oxygenated waters. The organisms found
below the Mason WWTP were chiefly in these insect orders
(Appendix D, Tables 2 and 3). The dominant organism at

stations SC-3, SC-4, SC-S was Chironomus which possess

 

hemoglobin that acts in both the transportation and storage

of oxygen (Walshe, 1950).

45

This survey showed reduced water quality below the
Mason WWTP which had substantial degrading effects on the

stream biota and general stream aesthetics for at least

2.5 miles. Lesser effects occurred for an additional

1.5 miles.

LITERATURE C ITED

LITERATURE CITED

Anon. 1973. Biological field and laboratory methods for
measuring the quality of surface waters and
effluents. U.S. Environ. Protection Agency,
Cincinnati, Ohio. 181 pp.

Basch, R., and J. Truchan. 1974. Calculated residual
chlorine concentrations safe for fish. Mich.
Water Res. Comm., Dept. of Natural Res. Tech.
Bull. 74-2. 29 pp.

Basch, R.; M. Newton; J. Truchan; and C. Fetterolf. 1971.
Chlorinated municipal waste toxicities to rainbow

trout and fathead minnows. Environ. Protection
Agency Water Poll. Contr. Res. Series 18050 GZZ.
49 pp.

Beck, W. 1955. Suggested method for reporting biotic data.
Sew. and Ind. Wastes 27:1193-96.

Doudoroff, P. 1952. Some recent developments in the study
of toxic industrial wastes. Proc. 4th Annual
Pacific N.W. Ind. Waste Conf., State College
(Pullman, Wash.). In Garton, R. 1968. Ph.D.
Thesis, Mich. St. Univ., E. Lansing.

Doudoroff, P., and M. Katz. 1953. Critical review of
literature on the toxicity of industrial wastes and
their components of fish. Sew. And Ind. Wastes
25:802-39.

Ellis, M. 1936. Erosion silt as a factor in aquatic
environments. Ecology 17:29-42.

Gaufin, A. 1958. The effects of pollution on a midwestern
stream. Ohio J. Sci. 58:197-208.

Gaufin, A., and C. Tarzwell. 1956. Aquatic macroinverte—

brate communities as indicators of organic pollution
in Lytle Creek. Sew. and Ind. Wastes 28:906-24.

46

47

Gordon, M. 1972. Animal physiology: principles and
adaptations. Macmillan Publ. Co. Inc., New York.
592 pp.

Hesse, J. 1973. Phthalate surveillance in Michigan waters.
Mich. Water Res. Comm., Dept. of Natural Res.
(Mimeographed) 9 pp.

Hesse, J., and E. Evans. 1972. Heavy metals in surface
waters, sediments and fish in Michigan. Mich.
Water Res. Comm., Dept. of Natural Res. 58 pp.

Hite, R. 1973. Michigan salmonid hatchery water quality
evaluation 1972. Mich. Water Res. Comm., Dept. of
Natural Res. 234 pp.

Hynes, H. 1970. The ecology of running waters. Univ. of
Toronto Press. 555 pp.

King, D., and R. Ball. 1966. A quantitative and quali-
tative measure of awfuachs production. Trans.
Amer. Micros. Soc. 85:232—40.

Knutilla, R. 1968. Regional draft-storage relationships
for the Grand River Basin. U.S. Dept. Int. Geol.
Survey. (Mimeographed) 1 p.

Lloyd, M., and R. Ghelardi. 1964. A table for calculating
the "equitability" component of species diversity.
J. Anim. Ecol. 33:217-25.

Mackenthum, K. 1969. The practice of water pollution
biology. U.S. Dept. Int., Fed. Water Poll. Contr.
Admin. 281 pp.

Mathis, J., and T. Cummings. 1973. Selected metals in
sediments, water and biota in the Illinois River.
J. Water Poll. Contr. Fed. 45:1573-83.

McKee, J., and H. Wolf. 1963. Water quality criteria.
2nd edition. Calif. State Water Quality Contr. Bd.,
Pub. No. 3-A. 548 pp.

Olive, J., and C. Dambach. 1973. Benthic macroinverte-
brates as indices of water quality in Whetstone
Creek, Morrow County, Ohio. Ohio J. Sci.
73(3):124-49.

Palmer, C. 1969. A composite rating of algae tolerating
organic pollution. J. of Phycol. 5:78-82.

Patrick, R. 1948. Factors effecting the distribution of
diatoms. Bot. Rev. 14:473-524.

48

Patten, B. 1962. Species diversity in net phytOplankton
of Raritan Bay. J. Mar. Res. 20:57-75.

Riley, C. 1972. A continuous-flow bioassy of Dart Con-
tainer Company. Mich. Water Res. Comm., Dept. of
Natural Res. (Mimeographed). 7 pp.

Sokal, R., and F. Rohlf. 1969. Biometry. W. H. Freeman
and Co., San Francisco. 776 pp.

Standard methods for the examination of water and waste-
water. 1971. 13th edition. Am. Public Health
Assoc., Inc., New York. 874 pp.

Steel, R., and J. Torrie. 1960. Principles and procedures
of statistics. McGraw-Hill Book Co., Inc., New
York. 481 pp.

Wager, D., and G. Schumacher. 1970. Phytoplankton of the
Susquehanna River near Bringhamton, New York:
seasonal variations: effects of sewage effluents.
J. of Phycol. 6:110-17.

Walshe, B. 1950. Hemoglobin function, Chironomus (Dip—
tera). J. Exp. Biol. 27:73.

 

Wilhm, J. 1970. Range of diversity index in benthic
macroinvertebrate pOpulations. J. Water Poll.
Contr. Fed. 42:221-24.

Willson, R. 1972. Biological investigations of Hughes and
Lake Drains and Brent Run, Genesee County, Michigan.
Mich. Water Res. Comm., Dept. of Natural Res.
(Mimeographed). 14 pp.

Young, 0. 1945. A limnological investigation of peri-
phyton in Douglas Lake, Michigan. Trans. Am.
Micros. Soc. 64:1-20.

Zillich, J. 1972. Toxicity of combined chlorine residuals
to freshwater fish. J. Water Poll. Contr. Fed.
44(22):212-19.

APPENDICES

APPENDIX A

PHYTOPLANKTON DATA

49

 

J

  

 

 

 

 

 

 

 

 

A m A o o o m N m o a nose. couus_—oa
o~.n F~.~ —c.m me.m o~.m mm.~ am.~ ~n.n mm.~ o~.~ oo.~ Amv mu.uto>.a mo.uoam
co.~ on.~ oF.~ m~.~ -._ mo.o em.o o no.o o o—.F nou~——ooapu ucoutoe
3.." e e e e e e e as e Re 2.232.; 352:
up.“ ~o.n a~.~ mo.~ om.v on.— cm.o Ne.~ 0 mm.— o meoogw accuses
no.o¢ ~..no oo.mo _~.mm mo.mm ~m.~a mm.mm mm.so _o.¢o m¢.wm mm.wo «noua.o ucousoe
5o 8: 2: 2: 8m 32 -= Sm «cm 82 ...8 33596 me Log: ~36.—
.N a n n E
m up mp up n m m .a» mace-opwmmmnp
m n m .an nan-mu
. n .am e3_c to
n o. a o— n m .am ecu :
n n .nm : nonwo
«cu-p. o—u
mp n m «seasons—us voeustuuuoca
ououuou vac ou
a a. amok
Aucoogu-o:—mv ouzgaocazu
m n m maoucaaep_c cuclecouuocs
a m o_ooooo oocmsLouwmcD
we o~ nu —~ nu w FN n .am macho oco
n .n« mmuu coo
m .a» ancwuootup:
n n n .3 £5 roan
9 .am Earuuu_00m
n n .am ssmsouuo_p
m 3 m m m n m .3 3382325
Ancovcuv ouxzoogo—cu
nu o. mm ~m ow pm m~ mu .n «cu-econ voc_sgou-s:=
a. moutucou anew-Lou c
a o— o— as o~ sap . no c— as on o .om aim” m
o— cu m 9— mp a _— I.an owWogmuaw
up —~ nu .e» noun co
. n— a .nm a acetaou
a... 8 2 3 2 n. 3 e. .3 :5: Soles.
asp onw «a. cop nc— so. RN. no» no. ohu am. .am o.mumum.2
n n n .3 SEE.
non mum own men can «on can mmu ace new «on .o« c—surraz
a o_ on as w a. nw ow a o~ .am cormwtm:
n— MN on up _~ o n
m n n~ o— o~
an um as «w n— we pm mm oN on mu
0. mg as aw o~ o— n— o
a m n
m up o— m a. o
mm a m a n . : F a
a 3 2 2 3 3 3 8 a .3 e so u
2 8 an 8 S e 2 2 2 a... 2 .3 3E
m m m m Mg m .au 3 one a
e 2 a an 8 e. e 2 a on 1.5%
on co. co mm on mo_ _m_ mm am No we .em mucus-:gu<
Amsououov acouxzaoutup—uu-m
use: coca coca .mwmw coca Neoasou urea mesmeou urea coax dun: come: as): com-z com-z o>on< ”couuouou on.»
8e: a... .52. 8.; 2.: :or .5321 :23 26$ :25: :28 28.. 8535 3. 8:. 553m .32
centowxo>o> ozone >a>o> um 00— ztoquOU .m
a e m o m m-> <.> « n N — ”acouuuum

._E\uEm.cooLo cu commutaxo sumo

.m~a_ .N x—sw .comuzuuz .zucsou canoe. .mcuman uco coma: Co xu.c_u_> ecu cu ozone za>o> use guano «Losuuzm c. nouuumcov nausea—nouzga .p< «pas»

50

.wrsoouoto acuucsou ~osuuo acutav uoc use coon v>.u~uu_o:u :. vo>tonno ~a_<o

 

~m.~
¢o.o
o

po.~
os.oo

c-p

~——
FN

FN
we

«o.~
w~.o
oz.o
pm.—
v5.5m
anew

mm

an

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wow
ow

aw

Np—

om—

ON—
an—

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op

own

nmo

cc
mm

m~
wc~

+

o~—
or

mn.~
cc.—

NV
oo—

mm
No

vm.~
n~.mp

”MM

mp

own

com
um

mp

amp

Ne

«n.~
no.m
o—.m
ca.—
mo.oo
mad

”to!”

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@—
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oa—

@—
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um.—
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pn.no
men

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up

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0mm

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no.0
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pwm—

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mm.mo
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mm.
m
mu
ow
n—
on
vv

pm
we

:3

"Km mun

.ov
n—

m—

 

seem
odor .L:

._e\mEmuc~0Lo cu commutoxo auoo

ease
asses

smug
ow...— as:

seed
ope:

uoox

mes_13ra

Fig:

 

voox
Lento:

m

xcoqsou urea

zo_om

crush xc>v>

nn>

acooEou Leno

m>on<

cuatp za>m>l

<->

.mso— .m tonewuawm coo_zuuz .xucsou sococz .mcumcoL uca cemoz Co xuwcuuu> acu

ease
P P030:

v

cu cuoto xa>o> was .xowtu on: .uuoto zo—_uz .xowtu usosouzm c. mouuumcou couxcoLQOuzze

as): came:
o>oaa wucatucm

xtmuw£Wpl.wll

N

nwz-m: sou.
sowtumos

p-91

 

cams: o>ob<

.ea ea_x

uses. eo.us__ea

Amy su.nras.e sarcasm

nsmueamso mo Loans: pouch

mouoppoOozu ucootoa
ua—m ucootoa
scootu ucougoa
”caucus ueoutoe

«canto-

vocLELuuouc:

 

.am mucosa crust
.am msoomn

.nm u:_toccoa

an o:u_m:u
nuuo—_uou_u

msoucwsop_u DacrsLouooc:
cwoouou moc.ELou~mc=

.nu ea

touo__womo
.am muum o~c<

Amcooguoo:_mv sunroocozu

naoucuso_uc voe_srouuo:=

vuoouoo uocLELououca

.am mssmomocaum

.nm satumsuoo
.nm m_um wmn

.nm Eswtoumo_p
.am usemuoorumzxc<

Aucootwv

cusseoropcu

“SESQEEESSS
muutucou vacaecouo c:

.nu qumo m

.am
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a 9.7:.
asses; sow

 
 

.nm aucormmoo_omw

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.n«

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.nm oppocourmum
.am ago

   

.om mocucuczu<
Ameououov uauuxzao.L-_p.uom

 

"co_uouo4
cozuoum

 

uncouuoum

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51

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APPENDIX B

PERIPHYTON DATA

52

.ovmunmm .mm .u.nom can

 

 

 

 

 

 

 

meom an Amomav muumEoflm ca mvocuwe an Umcflmuno mmSHm> .oocmflum> mo mflmhamcm umzuozu uou nwma maam> HafiOHuwuH¢.¢
some cw vmuaoflm uoz.

Hm.qnm om.me mn.mmm mn.mum o>.HuAMV some ocmuu

««mm.v u I H~.~ ~H.~ om.~ vm.m mn.v ~m.m no.0 Hm.m mm.oa «h.H ma.a mm.~ mbma .m .ummm
mo.v Nm.~ om.m Hv.~ mm.~ ~v.~ mm.n mm.» ~m.m wn.v vm.~ hm.o hm.a mN.H Hm.H mnma .NN .msm
mH.m cw.m mm.v no.5 om.h mm.m No.5 Hm.h -.o mo.o mo.o cmo.hm om.d mo.~ sm.a mhma .m .ma¢
m m < m m < m m < m m < m m < "mmamemm mung

:oHuomHHoo
m h o m a "codumum

.M HaunQOHOacu
1mm aasb .cwmwnowz .>ucsoo EmnmcH .xwmuo whoamumm

I

uo ~Eo\ma mm ommmoumxw mmsHm> HH< .mhma .m Hanewumom
cfl mxom3 o3» wuo>o nouooHHoo mono mcflvcmum couxnmwuwmuu.am mamde

53

TABLE B2.--Two-way analysis of variance for adjusted*
periphyton primary production in Sycamore
Creek, Ingham County, Michigan, July 25-
September 5, 1973.

 

 

Source D.F. Sum of Squares Mean Squares F
Station 4 53.6952 13.4237 4.303**
Dates 2 4.9169 2.4584 0.778
Error 7 24.9589 3.1198

Total 13 83.5710 19.4477

 

*Table adjusted for missing value for station SC-8
according to methods in Biometry (1969) by Sokal and Rohlf,
pp. 337-340.

**Significant at 5% level

TABLE B3.--Comparisons of differences between mean stand-
ing crop from Sycamore Creek, July 25-
September 5, 1973. Data expressed in ug/cm2
of chlorophyll a.

 

Station 1 7 8 6 5

 

Station Mean* 1.70 3.90 4.61 6.72 6.75

 

 

 

*Means not connected by the same line are signifi—
cantly different at 10% level.

APPENDIX c

ORGANIC SEDIMENT CONCENTRATIONS

54

.ucmwmz pm: mx\me cv vmmmmgnxm saweumu :

 

 

 

o.mm o.mm u m~.o u m.m o.p~ o.~ .>mo .uum N + cam:
o.o--o.¢ o.mm-o.p u o.mpump.o . o.muo.o o.ppuo.o o.~-o.o magma
m.Hm m.mm u m_.o . o.~ ~.v «.0 com:
mpm>m4
uczogmxumm
cam_;uwz mep
¢.mm¢ m.mmm ¢.~m m.o N._u~ o.mm - u.m .>mo .upm N + com:
m.om¢-m.uo m.onm-m.¢~ o.mm-~.m m.m-o.p ¢.Fofium.mu m.Hm-¢.m m.o-~.ov m.¢-~.p mmcom
H.Hmm m.NNH o.- N.m. ~.Fo 5.5F . m.m cam:
m.mom ¢.oom o.- .~.~ ".mm m.¢~ m.o o.m m - um
m.nmp ¢.om m.NN o.m o.mm ~.om N.ov m.¢ m . om
m.~m m.¢m m.¢~ ~.~ m.wF m.o~ N.ov m.~ s - um
m.mm~ m.mm m.m~ m.m N.N¢ 0.x, ~.ov m.¢ o . um
~.oom N.Pmp m.o~ ~.m “.mpp ~.m~ m.ov n.¢ m - um
e.~ofl «.me m.n— ~.~ o.mm m.op ~.ov ~.~ m u >
m.~m m.m~ ~.m o.~ o.- v.m ~.ov N.— < u >
m.mo¢ ~.mm~ m.mm m.m ¢.Pma m.- N.ov o.¢ v . um
m.ome m.o- o.mm m.m ~.mm~ m.Hm m.o _.¢ m u um
m.Pm_ m.~op m.mF w.P m.m~ m._~ N.ov _.~ N n om
e.—o— n.m~ ¢.ep s.F 0.“, m.m N.ov _.N F . um
uch now; meuPz mgzogmz Landau ESVEogcu assweomu owcmmg< "mco_unum
- .m—muwz

.«ucmwmz zgv .m¥\oe cw ummmmgaxm mmapm> PF< .mump .mm span .cwmwzuwz

.xucaou EmzmcH .:_mro mm>o> can xmmgu mgogauxm mo mucmevvmm souuoa cw venom mcowuugucmocou pupae xsmo: .pu «Fan»

55

 

 

 

 

~.n v.5 m.m m.n m.w ~.h H.n «.5 m.h H.h m.w an
odd mm «a mma mom mvw «av cow hum Hem oov Hwo
m.~ m.o m.ov o.o h.a m.oV m.ov m.m «.m m.ov m.ov name
”no moumanucm
H.ov H.ov H.ov H.ov H.ov H.ov H.ov H.ov H.ov H.ov H.ov vm.~a no mom
maxcwnmam
oouocwuoHnohaom
cam.o Hoo.ov Hoo.ov Hoo.ov mmo.o Hoo.ov Hoo.ov oma.o omm.o ov~.c aoo.ov ocopou0anu
ooo.o Hoo.ov Hoo.ov Hoo.ov aoo.o Hoo.ov Hoo.ov Hoo.ov voo.o Hoo.ov Hoo.ov cwuoaoaa
ooH.o aoo.ov Hoo.ov hoo.o mao.o Hoo.ov Hoo.0v mmo.o nmo.o muo.c Hoo.ov aoaumm
Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov Hoo.ov accumo
who.o Hoo.ov Hoo.ov moo.o mmo.o Hoo.oV Hoo.ov hmo.o nmo.o vmo.o Hoo.ov mos
cam.o Hoo.ov Hoo.ov moo.o «Ho.o Hoo.0v Hoo.ov NHo.o m~o.o mao.o Hoo.ov moo
mmm.o Hoo.ov Hoo.ov omo.o Hmo.o Hoo.ov Hoo.ov moo.o aaa.o moo.o Hoo.ov Boo Houoa
novwowunom conuoo
noun»: wouucwuoHnu
.om .vm .vm .um .om .vm .um .om m933 coon! mononucm .ux mmwx an umumEduum
moo: aaaon owns ado: Hound: uuco xonnmmom uHom zoaom auouosoo sown! «newuoooq
.u: we: .3 o3 .m «>34
m m h o m mn> ¢I> v n ~ A ”noduoum
.muacd undocuuu cw vomnoumxo aoaaa> mm .mx\ma

cw vouuoumxu nos~o> HH<

.mnaa .mN wash .cMOAcoaz ..oo EdnocH .cacuo >m>m> can xuouu oucEdOhm mo mucosauon EOuuon
0:» cw UGSOu Hao can .moumHmcucm .mH>cwnmfin omumcfiuoHcowaom .moofiowumom conuououv%n wouncau0aao uo acowuuuucoocoolu.~o mqmda

APPENDIX D

FISH AND MACROINVERTEBRATE DATA

 

56

HN ma nosau> ROUGH oacmona
av mm ma ms cm mausow>nocu no names: Houoa
Ha ma m 0H m mmwoomu no noses: ch09
N Anamosa xomuuv oxwm cnwnunoz
N Ammonosanm mznmummnonzv anon susoeomnmq

v moonm

om mN vH

HON

 

 

n «a Amnnuuomdn mounamownadv ummnxoom
n n A Amsmonnnm .3 oommcnxm§m
m NH Amsaaocowo mNEomunv gunman» coonu
m muono

 

5H N

 

m m handmaonm mwamamUEmmw soccua onwnuom

N Ammosuaonwno macowaemuoz. nmcwnn cwuaoo

Annuscnoo .m nmcwnu GOEEOU

n.@@ on onuozv mnocwnm

N Ausasuonuu mwmumoacwnmv moon omocxooam

N AHENH unQEDV soGGMEUsE Hunucoo

mN Ausauasouaonum moanuosomv £550 xwono
N
N

 

 

 

 

 

Assuasnwmo .mv nounou toncnmm

«adnmmm «souwoonuu. nounuv ancnon

Aouoaaoua canonom. nuance moan on m
N moono

 

HIDVNHIn 5

HH IQGHU

xlhnnkumc .HV voocaaan koaaou
nomads manSHmvo N vuosaasn xuoam
N NH ma \hwcomnoeaoo msEoumououv nwxosm mung:
Aucaownuwc Eunwoucumwwv noxosm wmocmom
H A.mn naouwoxoz.,n¢xosu canon mm
H msonu
H mmmNU

(Dav
H

 

HHM V

 

.um xaaon .um cone dawn .vm anon .um nouns: oocmnucm .om mung Andean onunuconou can coeeoov
Benn Bonn Eonu Eonm xnmuwfimu .m Bonn “cenumooq monowmm swam
Edonumm: swmnumma Emmnummn Emonumaa Eonm Emmnummo coNumum
Emmnumma

 

m h w m N a uncowuuum

 

. . .23 .n nonemumom
savanna: .oo EdcmnH .xuono onOEdo>w Eonu weenumum Uouomaom um manzmwuonuooaw >2 cwuouaaoo undo scan «0 unaEEdmln.Hn wqmda

s7

 

 

N
N
n
N
o—
n N
o
N
_
_
_
ON 2 G o: S
p v o. —
p
N Z N n.
v —
oN _ m 8 N
a N N o
—N ON 3 Na 1
p
—
_
on R o N
o o n
N N N
N —
s—A o— cm 8» mm
o m — ON
: p v m p
n — —N
n
m
m N n n
— —
_ N
o. p
N
N_ N
n p m
N
p
_
N N N N
o. N
.3. .2. ll? 3 two.“ -
3o: 3.2. 3: :o: 35::
,E 8:
2:1-Nt1 w I “Illldll|4.f

.22 .m-v 59.02am .5625:

.

. :53 .559: .Efo :3) 0:. £35 on: 5.35 .62; .395 9.9559 £0... «83323 2.23.2.3 c. 5.3 3:233:33? :22...

 

N
a
o — n
_
N N m
— N
N n o _
— — o p
n
_
m n o
_
K a N _ p
n
2 c n—
. n
_
—
_
N
n o. N o
n ON
_
3 N N
3
n n 3 Z N
N. v o.
o. p m 2
m.
n. o
—
N
N
—
N N n 8.. S
o
.2 .3 .8 .3 3:... 1.2::
.393: 53:28 LOSSES Zulu: 10?: :03: good
{so too .u.‘ 8. 3.3.5:“.
lo—vm use: .N..o...¢..uu.
o. a; £43.. a q - .

NS . m:
:6:
£3.53:

N.

87

N—

I... mam.

:ti. 2...;

.Huummmnu..-mqmaqum

NH—Hflwwa, .. oem flwfimuflzedm
33:5: .35....3: 528:8:

- wmmtnpmm

.nn vaueOOufiEchm

3 33%.

— .55: 7.3.3.

 

 

.o..«....oHn.
ouo'uwov:3 swinger
.3 New amt/3H1
kwummmwwm.

“gnaw 3F. or NINSVI

Warm—Huffltwo

navi—Lo

_ «3.3 28333
:25 aztwfini

.mm «Viewek

.am wahmgfo

—N Sue—aim”... “hmwwdm—m

_ .3 de§

0325.34

.3 .mvcmwfl
...._._..a-u ..~_,.comm.uw .s.xsvo

.3 EbE...E

.ou cEmccgNm

gbwwmsmm aucuflwwllm
Sheaves.“ couW'rflfluMMn
39...“...3.
flcfimmvmfl
«a SEEM
.3 fldwzflu
ON .3 2:3
33:33 Couaotflfiiu
.3 «73:18.3
ASNCoconwlstwuuoum—a
m.— TJ> .
a a mwwclbss
.3 3.3956
833

 

 

 

 

mN

 

 

 

 

87 .3 “3.35

ml. u “wanna UL
253.: «33> .5
00:0. uLooPSuK
.3 ca ET
.3 coast.
«395:. IsHBImLH
«:33»: “330ch
.3 337:0
I333. 58 flu
32: .K
mun-2: 18:!
30.! .u E: .mREmaJ
i? I. one?!
— alga 3:0: 33
N 32223.: nZQoo Lu
2.? _ 2309.“
33m «SwooozufuoN
7059...: 3:632.
Ailing-o 9226: 3358.8
vast-:33
49:95an sic—pooh...

 

 

 

 

 

 

N-NS
I
l

 

 

    

— ILIL_'——lt——— hummus; uuukuumnhmmuu “but.

_|L this

fi-hfi-fi-D— “It“.ILLI—ILO—ILIL I‘m“. .—

.—

 

.comax ”comoluob was:
2.84 utzcflum
.3. 938

_ “c0230

magnum
cucoto_on

u... 30..»

58

 

2

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l-NUD

 

 

one:
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I.

 

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out
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on

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n. m—
c

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NooL-x soc.~ucou
“Lao

110m

fl 9,

N—'

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58.3.8...
“Lao
26.2

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.3
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an

3...:
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2: 8—

a-

331

count 209.
3:755
P333 .N

N

N, . m:
8.;
5833

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coast
u,oa<
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«co—unifies woman 5 uulvqui “3 .
:3 59: :2;

$38» 2313 No 53.52

“03!; 3.33.3: No L095:

«03!: “coho—3c. No .3952

$38.. No .305: ~33

 

.3 £43:

gone 23825,.de
.nn N:m€3»..5.cbch
.3 3m»:u«...1npq,.l.\élfi
.nm an.wa.uogu

.3 M1.,vwm..w..m.
$4.1... 5.5mm 2..
:Echg

.0a auaoruwrmh

.ou MflDMaglufea
.3 Eff“...

.n» alvdvflccowmbdd
.3 EUquoguE
.3 Ztnflafifl-
:93... .c agavmflx okra
ow n,n-.c_u

.Pl..%.+..my9m
b

 

 

 

 

 

.n» «gc8_630
.aa mmmywflu_mw
33:33 .3..«..fi..f.6

0.. 35. N05 3
.3 “uranium“.ab
An:.o..a. thistnb

 

numhumuuuumumukumuuubr—uuuu

:83: 83899.6
.3 :33
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.3 waif;
26...._g
.9 3..th
.am 393an
.nm giggle
.2. “£3qu
.3 383%
,lé. are
“‘03 :a 233:1
7.0;: .2306
.3 wackurmmt

 

 

 

 

khkhhkhkhulfi

 

2593:“ 3mm c9nuleu
.cn SZKdS-N
.n» «a. N

 

.ou «nu->OLo x
.3 S 31......
.9. «:c.Amu

.n. -. ounhh a
.o« c—gqumuuu
commuu.uxn
.c .L_mmb

.nu ’auaoc.n

.3 3335MB
.3 «388

Ana—uooe fiance—cu

¢ a 3 wmu .MK
.3 “ammucaufio
vet-6 - www.mwlm

.aa 05 «mm; x

 

 

I‘M-KKKKKEKKKKKKM-Kkh

 

 

 

.sfimmm. m

L5: .,..

99.0 attic wriswwnfl x

.3. XWN¢Q935cfl

.ou TIN-wad
32:33.35 30305....»

 

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2.8.533 aria! «33...
3:28.“ 3:310»

 

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232.383 .8 .21.—

59

23:39:“. 3.003 c. 33.2.. yo: .
982 “4.8. 9...... 932 m 38 $8 «.2: «.32 m 8.. $2 8. S..3:.u\i:$eso .o teal... .33 3.1....
m. 2 8 S 2 n. m a 2 S 8 5:33....33 .o .85.. .3...

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1.. an 2. 2.. o. a o. 3 03 m: 3 2. .58.. la: Lafimwfi

o. 2 o. ~n no a: o. . ~n 9 1.53193}.

0 a».

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S o. 3. S 2 arsoxwa...

a N. 3 a. o. o. 3 flamflgmflw:

o. 2 3 3 .33 m1!

8 3 .o S 9 ~. 8. 2 2 2 3 2 .3 yum:

.9 3. z 3 2 2. o: m: o. 8... z... o; 2. 3. a. a: n: =2: 5:354}...

2 2 .o a 2 o. .2 L35“.

9 0- .An arduovfibuflmmndoa

up 3 a. o. .3 «Hammawgflfdm

o. .3 aufl..;,._..mwm

o— u. 7.13:.

an gflaumflflm

.3 3...|.,,......_a4flm

a. 3 .2 $5593

9 .3 wake“... find...

3 6’ 33.3133

o— 3 o .v ..vu..u....d.u

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a 8: .8 .2 8. 2 ,2 3.9m...

man can 3 “M .nu'aQGcZmela

2 «audio. 3 El

~u .. .. .n a. o. o. n.. Q. ~. ._ ”_“w“.._ «w

9 3 o. 2 a. .3 an C

o: 2 a. nn 3 my. .3 .Hcmqgfl.

2 o. . ,

3 2 .8. o. a. 3. at o.
.2 a. o. 8 8 8 8. 3. 39 3.. .8. .t .c a o. o.

 

 

 

 

 

 

5’32;

 

 

 

KKKFBK‘KKKKLKKbKKKKK&&&&KKMD-hb‘LI-&KK

2 2 3 a o. no so no 0. '9 3n 3 9 3 o. .33
303. osElo 8.
Cu :— 3 p. 3 A . 09v. . .caGgu
«3.33
3 3 3 2 03.0.3.
:0. 3.0.39
2 .3 3:5 .Imwm
8' a no a: 3 on :— 3 no manage.“ Mac 1.34ch
no 2 NH 3 .3 23.51.30
2 goalie?» “Ncazmri.
7338. ZZEELS
o— .3 at. 349.50
2 3 < .38 mag
.L. . a
2 gag.“
.3 an as N: 3: Z: 2 .3 a:
an Na 3140:... .m
3 nu .3 no 5 co— .cu_...u.,. .m
a 8. 2 a .. 2 2. o. 3 .3... add 53
. o. .9. a;
scan as 32 an: 82 can 2: co. 3 a: Na .0. o. A .3 Ev a .!95
.0 a v3 3.3093 L»
2 gin. an...”
32.2.: $5.52: :3331
o. .3 3.25.5...

92

 

I‘KKK “Kh-

 

 

Khhhlh‘l-ILK—

Kb

3 2r: 3...... I3.
2 o. 5.. 9 R $.13” - 3
o. .3 12$.”
3 no 2 2 o. :35;

30.2.... 30:28.: 3253
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«a an o. no a. Iaflg... o:- m

a. 823.. 8
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2 Na '2 an 3 o— '0— ;w .3 2C;

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a: 7.3.3 %
no: u .—

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2 83.53:.

2.83.... 35.8....

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. < a < . ¢ . c. u < n « a < . ‘ n < . ¢ o < 0.93

 

 

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3.8 .3 oo— nsuulu .u Satan: cow-i552 3.3.0.0..

 

. o . o .9. m u n ~ 3.. . $83.: -.

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60

 

 

 

OH HO ON mm «m NH N O «N NO ON coHuuumxHHm. xwocH oHuon
O0.0 H0.0 O0.0 N0.0 Om.O O~.O O~.O O0.0 m«.O O0.0 O0.0 :oHuuum\Ho. HuHHHnmuHsvm
O.~ O.~ O.~ H.m O.m m.H m.O O.H O.~ O.m m.m coHumum\HmO muHmum>Hu uoHoomm
O.~ m.O m.m O.v O.m 0.0 O.H O H.mH m.OH 0.0~ uHmncH>chH
umnuo mo umnfisa unwoumm
m.OO H.O 0.0H O.mm O.H~ 0.00 m.OO 0.00H O.mO m.HO 0.0m mwvae «0 Hones: ucooumm
0.0 0.00 m.mv m.mm O.~O O O O 0.0 0.0H O.H moHmeHvomo mo amnesa unwoumm
O 0.0m ~.Hv 0.0 O.HH 0.0 O O O.” 0.0H 0.0H mmHHuaua no Hogans uswouom
Ow HOm m.OOH m.~HH m.OO~ OH m.OO O m.Om~ OOH m.OO~ ~e\numzuo no umnsaz
m.OmmH m.OO~ m.oom HOO m.VOvH «MON Once m.~omm NONH OOO m.OOm exnomuHs no Honenz
O OOmm m.OO- m.OO~H m.~OHv O O O ONH m.mOH OH ~a\momHumHouuo no nonesz
O «OO~ mmON m.OO~ MOO O O O OO m.mvH mOH ~5\moHHuaue no umnesz
m.mOmH m.mmOO m.HHom m.OOm~ m.ovwo OmON m.~HOv m.~omm m.OOOH «mmH MOO e\m5uH:mmuo
mo am a wouueHuuu
0.0~ O.mn O.m~ 0.0~ O.o~ O.mH 0.0v O ¢.OO ~.OH O.m~ nuHoomm uosuo ucoouum
n.nO O.mn O.mv 0.0m n.mv 0.00 0.00 0.00H m.mv o.om O.mm uwHoomu oOcHe unmouom
0.0 0.0N 0.0~ m.vH 0.0~ O O O O.mH n.OH 0.0H uoHoomu aHuchvou unmouom
O O.OH O.OH H.O O.OH O; O O O.mH O.OH 0.0H «33% 3:3 £588
O O n O O N O O O m m uoHoomm uosuo no nonsaz
HH O O OH OH OH O O OH OH HH muHooaa omcHa «0 nonaaz
H v v v w o o o m n w mmfioomm mHmmwmooo mo uonasz
O N ~ w m H O O n O H 253% its. no 333
mH on on mm on nH m m MN aw om ouHooam no nonanc H6909
.um .vm .nm .nm .um .cm .vm max: mocmuucm O~H .m.o .um mme uncoHumooq
omen >HH0H mwua uHom umHHchm humus: HHmaom :Omoz mumumewo Eouu an sown: coHuoum
.u: och 30Hmn .m adwuummo o>on<
.uu OOH
O O O o Hnuz m O m w Huuz H "acoHuoum

 

.man .m umnfiuumwmumm hHsh .comwnowz ..ou adnwaH .mxoouo

us: van .30HHH3 .0uoeuo>m ca voUMHm muonEdm oumuuunau HnHUHuHuuu choouuovmmm scum «and wumunwuuo>caouuua mo muueaswul.vo manta

61

coflumum\awv mm mEMm msu uou mH mO£BO

 

 

mO.N mm.m mo.m mm.m OO.N mv.H mm.o NO.H HO.N mv.m Om.m «meQEmm

mo va mmmum><
mm.m mv.m oO.m HO.N mO.m mm.H mv.o 0O.H Hm.m mm.m mm.m m mHmEmm mo fimv
oo.m oo.m om.m Om.m mO.m mo.H mH.o OO.H mm.m Hm.m mH.m ¢ mHmEmm mo fimv
m m O o 02 m v m N 03 H "cowumum

 

.mOOH .O ”mnemummmumw OHOO .OOOOnon .Ouasou
EmsmcH .mcflmcmq can gown: mo huwcwow> 0:» CH mxmwuu was new .30HHH3 .oHOEmomm ca mmumuumnnm
Hmwowmwuum wvcwolumummm unasouwo co cmuomHHou mmumuamuum>cflouoms «0 Hey >uwmum>wv mmflommmll.oo mqm¢e

 

 

O.NH 0.0H 0.0H O.NN O.Hm O.O 0.0 0.0 0.0H 0.0H O.OH cum:
OH OH OH OH Om O O O OH OH OH O mHmamm OH mmHummO
OH OH OH ON OH OH O O OH OH OH O mHmsmO :H mmHommm
O O O O o: O O m m o: H "coHumum

 

.OOOH ~O “answummmumm OHOO
.cmvwnoflz .uucaoo EdnmaH .mcflmcmq can cemmz mo wuflcH0H> onu cw mxmmuo on: cam .30HHO3

.muoemomm GO mmumuquSO HMHOOMHuum Occmonumummm ustouHo co omuowHHoo mwfiowmm mo umnEsznl.mo mqm<9

62

.Hm>ma wm um uCOHGMMHU >HucmoflmwcmHm mum mcHH mEmm may >3 Uwuomccoo uoc mammZO

 

 

 

mv.m vm.m mm.N wh.m hh.N HB.N o®.N om.N mv.H Nv.a mm.o AUV mmmum><

03 H o m 02 N h m m m v coflumum

.MOmH .m umnawummmnmm aHsh .cmmwnowz .>HCUOU EmsmcH .OCHmcmn can sown: mo huwcwow>
on» GO mxmmuu as: wad .30HHH3 .muosmomm ca mmumuumnsm HmHowmwuum mvcwouumummm co
cmuomHHoo mmumunouum>cwonome mo mmwuwmum>wn mmfiowmm cmmzumn mmocmummuwv mo mcomaummsboun.mo manta

.Hw>wa wm um ucmumMMOw aduCOUOMOcuHm mum mcflH OEMm ms» an omuomccoo uoc mcmwz«

 

 

 

o.mm m.am o.mH m.wH m.ma m.mH m.mH O.NH m.m m.v o.v mmflowmm mo Hmnfidz cmmz

O 02 03 O N O H O O O O coHumum

.MOmH .m umnswumwmnmm OHSh .cmmflnowz .wucsoo EmnwcH .mcwmcmq cam comm: mo muwcwoa>
0:» CH mxmwuu on: can .30HHH3 .muoemowm GH mmumuumnam Haaowmauum zocmaluoummm co
nmuomHHoo mmHommm mumunmuum>cflouome mo Hmnsdc came cmmzuwn mmocmuwMMOv mo mnemfiummeoonu.Oa mqmde

"“‘i'fliflifllflmjlwfl‘lmflWET