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THE EIFFECTS OF LAMIPREY LARVICIDE ON THE EOTTOM
FAUNA AND PERIPHYTON OF THE CHOCOLAY RIVER,
MARQUETTE COUNTY, MICHIGAN
By
Robert C. Haas
A thesis submitted in partial fulfillment
of the requirements for the degree
of Master of Science
in Fisheries
May, 1970
School of Natural Resources
Department of Wildlife and Fisheries
The University of Michigan
Committee members:
Dr. Frank F. Hooper, Chairman
Dr. W. C. Latta
Dr. Justin W. LeOnard
ACKNOWLEDGMENTS
This study was supported by the Institute for Fisheries
Research, Michigan Department of Natural Resources. I wish to
express deep appreciation to Dr. Frank F. Hooper, chairman of
my committee. Thanks also go to the other committee members,
Dr. W. Carl Latta and Dr. Justin w. Leonard for their very
helpful advice and suggestions. I am indebted to J ames Ryckman
for help with the statistical analysis and to James Merna, Burt
Wagner and Martin Hansen (now deceased) for the field collections.
Mr. Robert Braem, fishery supervisor, and the staff of the Bureau
of Commercial Fisheries were most helpful in providing the
larvicide treatments. Finally, I wish to thank my wife, Pam, for
her assistance with the first draft.
ii
TABLE OF CONTENTS
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . .
Study Area
Sampling Areas
METHODS . . . . . . . . . . . . . . . . . . . . . . . . . .
Bioassays and Application of Larvicide
Bottom Fauna Sampling
Periphyton Sampling
RESULTS AND DISCUSSION . . . . . . . . . . . . . . . .
Bioassays
BOttom Fauna
Periphyton Results
SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . . .
iii
iv.
Vi
3
13
14
15
19
21
25
27
50
62
64
Table
LIST OF TABLES
Some physical and chemical properties of the
water of the East Branch of the Chocolay River
on September 20, 1965, at the TFM treatment
site . . . . . . . . . . . . . . . . . . . . . . . . . .
Temperature, water stage and current velocity
for the study section of the East Branch of the
Chocolay River in 1965 . . . . . . . . . . . . . . .
Some physical and chemical properties of the East
Branch of the Chocolay River, and the biological
activity of the larvicide in this water as determined
from pre-treatment bioassays . . . . . tº º º º tº º
TFM concentrations with variations and tempera-
ture in the East Branch of the Chocolay River
during treatments . . . . . . . . . . . . . . . . . .
List of macrobenthos taxa in the East Branch of
the Chocolay River in the fall of 1965 . . . . . . . .
Genera and relative abundance of macrobenthos
selected for statistical comparison from before
treatment samples . . . . . . . . . . . . . . . . . .
Changes in abundance for each genera of bottom
fauna from riffles over the specified time interval
calculated as the percent of the former abundance .
Changes in abundance for each genera of bottom
fauna from pools over the specified time interval
calculated as the percent of the former abundance .
F-values for sources of variation from three-way
analysis of variance comparing riffle bottom
fauna samples
iV
Page
22
24
26
28
34
36
39
42
45
Table
10
11
12
13
F-values for sources of variation from three-way
analysis of variance comparing pool bottom fauna
samples . . . . . . . . © Q & © tº º O Go O Q © e e º O e
Current velocities at periphyton areas in feet
per second . . . . . . . . . . . . . . . . . . . . . .
F-values for sources of variation from four-way
analysis of variance comparing periphyton
sampling areas . . . . . . . . . . . . . . . . . . .
Mean periphyton weights in grams per square
meter from two sampling periods innediately
before and after each treatment with TFM . . . . .
Page
46
51
59
60
Figure
LIST OF ILLUSTRATIONS
Page
Study section of the East Branch of the
Chocolay River, Marquette County,
Michigan . . . . . . . . © tº 0 tº e º e C o e º O e o O e 6
Experimental riffle bottom fauna area (A) showing
how string was used to delineate sampling stations . 9
Experimental pool bottom fauna area (C) . . . . . . . 9
Lampricide treatment site showing turbulence
created by current deflector used to mix
chemical . . . . . . . . . . . . . . . . . . . . © e o e 12
Grid method of selecting one-square-foot random
bottom fauna stations at stream sampling areas . . . 18
Temporal arrangement of riffle bottom fauna sampling
schedule with mean number of benthic organisms per
square foot in parentheses . . . . . . . . . . . . . . 30
Temporal arrangement of pool bottom fauna sampling
schedule with mean number of benthic organisms per
square foot in parentheses . . . . . . . . . . . . . e 32
Graph of periphyton dry weights over the entire study
period for the pool areas. . . . . . . . . . . . © C gº 54
Graph of periphyton dry weights over the entire study
period for the riffle areas . . . . . . . . . . . . . . 55
Vi
INTRODUCTION
The sea lamprey (Petromyzon marinus) gained entry to the
upper Great Lakes in 1829 upon completion of the Welland Canal.
By the 1930's the sea lamprey had established itself in Lakes Huron
and Michigan and a short time later in Lake Superior as well. This
parasite thrived and by 1950 the commercial harvest of lake trout
(Salvelinus namaycush) was down 95% from that of the 1930's
(Eschmeyer, 1957). The disastrous decline in harvest of the highly
valuable lake trout prompted cooperative research and lamprey
control programs between Canada and the United States as early as
1946 and a treaty for joint action was signed in 1954. The inter-
national collaboration resulted in life history studies (Applegate, 1950;
Applegate and Moffett, 1955; Hile, 1957; and others) which led the way
to a lamprey control program based upon the vulnerability of adults in
spawning streams. Various mechanical and electrical weirs were
used to block lamprey migrations both up and downstream; but high
costs, time required to reach desired results, and flood conditions
made weirs in practicable (Applegate, Smith and Nielsen, 1952;
Erkkila, Smith and McLain, 1956). It was found that larvae of the
sea lamprey, called ammocoetes, live in the spawning streams for
about 5 years before maturation to the parasitic, lake dwelling adult.
Hence, it was possible to treat the spawning streams with a toxic
chemical, kill the larvae and eliminate several generations before
they became parasitic.
Applegate et al. (1958) found a differential toxicity between
fishes and larval lampreys for ten halogenated mononitrophenols.
All of these compounds are more toxic to lampreys than most other
aquatic Organisms; however, one of them, 3-trifluormethyl-4-
nitrophenol (TFM), met the requirements more closely and was
selected for field use (Applegate, et al., 1961). Laboratory studies
with TFM (Applegate et al., 1957; Applegate et al., 1958; and
Applegate et al., 1961) established that this chemical is acutely
toxic to larval lampreys at low concentrations (2–3 ppm) and that at
these concentrations, it is non-toxic to other fishes.
Stream treatments with the lampricide are timed to remove
the lampreys before metamorphosis. Most of these streams contain
valuable resident fish populations and spawning habitat for Great
Lakes fishes. Although numerous trials demonstrated that concen—
trations of TFM used in stream treatments (2-4 ppm) had little or
no direct effect upon the resident fish, the possibility existed that
this chemical might eliminate some of the stream invertebrates and
algae and thus remove a part of the fishes' food supply.
In some instances, representative stream invertebrates
were included in the laboratory analyses and early reports showed
that crayfish and insects were not affected at the concentrations used
to eliminate lampreys (Applegate et al., 1961). Field Observations
of actual stream treatments likewise did not indicate gross Stream
invertebrate mortalities. However, before the present study, an
intensive investigation of the effects of TFM treatment upon stream
invertebrates and algal communities under natural conditions had not
been roade.
In the fall of 1965, the East Branch of the Chocolay River in
Michigan's Upper Peninsula was treated with TFM to determine the
chemical's effects upon stream bottom fauna and periphyton. The
purpose of this paper is to report and evaluate the short-term effects
of the larvicide on these communities in the East Branch of the
Chocolay River.
Study Area
The East Branch of the Chocolay River is located in Marquette
County, Michigan. The Chocolay system drains approximately
94, 000 acres of 1and and flows into Lake Superior at Harvey, Michigan.
The East Branch is 8 miles long. It has old beaver dams and ponds at
the headwaters. The river is surrounded by moraines covered by
mixed northern hardwoods and conifers. Bottom soils of the East
Branch consist mainly of sand and gravel with occasional rocks and
boulders. Water color varies from light to dark brown due to organic
compounds added by the beaver ponds at the headwaters.
Portions of the stream flow through rolling farm country
where bank erosion is quite noticeable. A biological and physical
inventory of the drainage system by Galbraith (1954) indicated that
temperatures in the East Branch might occasionally exceed the level
generally considered lethal for trout. These high temperatures
were attributed to lack of cover along the stream banks.
The area studied (Fig. 1) consisted of 1 1/4 miles of
stream in Sections 25 and 36 (T 46N, R 24W) of Branch Township,
Marquette County. This portion of the stream is about half shaded
by bank vegetation with a typical habitat of pools alternated with swift
running riffle areas. The width of the study area varies from 6 to
13 feet. The average current velocity was about 2 ft/sec in the
riffle areas and 1 ft/sec in the pool areas during the study period.
The substrates of the riffle areas consisted of rubble and gravel and
the pool areas were gravel and sand. Water temperatures during the
TFM treatment period varied from a maximum of 59 F on September 18
to a minimum of 37 F. On October 15.
Fish species resident in the East Branch of the Chocolay River
(Galbraith, 1954) were rainbow trout (Salmo gairdneri), brook trout
(Salvelinus fontinalis), blacknose dace (Rhinichthys atratulus), redbelly
dace (Chrosomus eos), mottled sculpin (Cottus bairdi), brook stickleback
(Eucalia inconstans), and central mudminnow (Unbra limi). Migratory
rainbow trout, brown trout (Salmo trutta) and sea lampreys are barred
from the study area by natural waterfalls downstream.
Figure 1. --Study section of the East Branch of the
Chocolay River, Marquette County, Michigan. The
experimental and control riffle bottom fauna areas are
designated A and B and the experimental and control pool
bottom fauna areas C and D. The experimental and control
riffle periphyton areas are designated Y and Z and the
experimental and control pool periphyton areas W and X.
Sec. 25
Larvicide Treatment
Sec. 26 Point
Ared Z
4’ ºr
Sec. 35 | E - - - - - - - - - -*...*
• Location of Bottom Fauna and Periphyton Sampling Areas
Figure 1

Sampling Areas
The location of the sampling areas for bottom fauna and
periphyton are shown in Figure 1. Four bottom fauna sampling
areas and four periphyton sampling areas were selected; two riffle
areas and two pool areas for both periphyton and bottom fauna. One
area of each type (riffle and pool) served as an experimental area
and the other as a control area. Control areas were located upstream
from the point of treatment with TFM and the experimental areas
were downstream. The corresponding experimental and control
areas were selected to be as similar in physical and biological
characteristics as possible. Two of the bottom fauna areas are
shown in Figures 2 and 3.
Areas A and B were the experimental and control riffle
bottom fauna areas, C and D were the pool bottom fauna areas.
Likewise, x and Z were the experimental and control riffle
periphyton areas while W and X were the pool periphyton areas.
Riffle bottom fauna sampling area A (Fig. 2) was
characterized by water depths of 3–11 inches with a substrate of
rubble and gravel overlying sand. The percentages of materials
in the substrate ranged from 50% rubble and 50% gravel to 90%
rubble and 10% gravel. The average current velocity for the
treatment period at A was 1.4 ft/sec.
Riffle bottom fauna sampling area B was very similar
in physical properties to station A. Area B was characterized
Figure 2. --Experimental riffle bottom
fauna area (A) showing how string was used to
delineate sampling stations.
Figure 3. --Experimental pool bottom fauna
area (C).
Figures 2 and 3

10
by water depths of 6 to 11 inches and also had a substrate of rubble
and gravel overlying sand. Composition of the substrate ran from
40% rubble and 60% gravel to 90% rubble and 10% gravel. Average
current velocity for the treatment period was 1.4 ft/sec.
Pool bottom fauna sampling area C (Fig. 3) had water depths
of 8 to 28 inches and a substrate of rubble overlying gravel and sand.
Rocks covered the sand and gravel in varying degrees from 50% to
100%. Average current velocity for the treatment period at area C
was 0.5 ft/sec.
Pool bottom fauna sampling area D was characterized by
water depths of 10 to 24 inches with a substrate of rubble overlying
sand and gravel, covering the latter two from 80% to 100%. Average
current velocity for the treatment period at area D was 0.7 ft/sec.
The lampricide was introduced into the stream approximately
midway between the experimental and control sampling area (Fig. 1).
This site of introduction is shown in Figure 4.
11
Figure 4. --Lampricide treatment site
showing turbulence created by current deflector
used to mix chemical.
12
Figure 4

METHODS
Current velocity was measured with a Gurley current meter.
Each velocity measurement was determined from three separate
readings of the meter for 40-second periods. Current measurements
were made within 6 inches of the stream bottom at all locations.
Near the treatment site a staff gauge was placed in the
stream bed so that the lower end was always below the minimum
water level. This gauge, graduated at intervals o: 1 foot and tenths
of feet, was used to measure the height of the stream water.
A subsurface maximum-minimum thermometer was
employed to register upper and lower stream temperatures
throughout the study period. This thermometer was located in the
middle of the stream near the treatment site.
Chemical analyses of the stream water was conducted
several days prior to the first treatment with TFM. Procedures
for these determinations were generally those set forth in the
ninth edition of "Standard Methods for the Examination of Water
and Sewage."
13
14
Bioassays and Application
of Larvicide
Applegate et al. (1961) found that the amount of TFM and the
time required to treat a given stream cannot be determined by chemical
analyses of the water, therefore, pre-treatment bioassays have to be
conducted for each stream treatment. Bioassays were made from a
mobile laboratory using Chocolay River water. The methods used
were essentially those described by Applegate et al. (1957). Test
animals were placed in containers and subjected to various levels of
TFM to determine the minimum lethal dose for lamprey larvae
(concentration killing 100% of the test lamprey larvae in 24 hours),
and the maximum allowable dose for fish (concentration killing 25%
of the test fish in 24 hours). Larval brook lampreys (Ichthyomyzon
fossor) and rainbow trout were utilized as test animals. The
bioassays were adequate to determine the minimum and maximum
allowable dosages of the lampricide. Test cages that contained
specimens of the brook lamprey and rainbow trout were also placed
in the stream well below the treatment point. Their purpose was to
determine whether or not the concentration of TFM was sufficient to
kill lamprey larvae but not trout within a 24-hour period.
The efficient application of TFM requires a highly accurate
and controllable pumping system. The system as described by
Applegate et al. (1961) was arranged to feed a concentrated stock
15
solution of TFM into a pipe containing a stream of water drawn from
the river by a centrifugal pump. Stream deflectors were positioned
at the treatment site to mix the TFM with the river water (Fig. 4).
Metering of the diluted lampricide through a perforated pipe aided in
mixing the chemical with the water.
Successful treatment with lampricide requires a precise
method of analyzing the treated stream for TFM so that the needed
concentration can be maintained. Accurate measurements of the
amounts of TFM were made by colorimetric analysis based on the
natural yellow color of the nitrophenols. This method was described
in detail by Smith, Applegate and Johnson (1960). These analyses
were made at three stations below the lampricide feeder unit. These
stations were located about 200 yards downstream from the treatment
site and arranged perpendicular to the flow with one on each side of
the stream and one directly in the middle.
Bottom Fauna Sampling
The four bottom fauna areas were sampled during September
and October of 1965, to evaluate the changes in abundance due to the
effects of TFM treatment. The riffle and pool areas were separated
upstream and down from the point of treatment by approximately
200 yards and the same areas were used throughout the sampling
period.
16
The area boundaries were first marked with permanent
stakes driven into the stream bottom. These boundaries were at
least 1 foot from the stream shore. Then each area was divided
into numbered square-foot sections by a grid as exemplified in
Figure 5. To locate each sampling station, a string was stretched
around the perimeter of each area and a tape was used to measure
the coordinates for each sample within the stream area.
Ten random square-foot samples were taken from each
area during each sampling period. Numbers for sampling stations
were selected from a table of random numbers. The square-foot
sampler was used for all collections of bottom fauna. According
to Welch (1948), it is especially suitable for collecting macroscopic
organisms in stony and gravelly stream bottoms which possess enough
current to hold the net open. Stations that fell partly or wholly on
large rocks or logs were rejected, as were those stations that had
been sampled previously.
The corresponding experimental and control areas were
sampled on the same date. The bottom fauna samples were placed
in shallow enamel pans and were picked innmediately while the
animals were alive. The rubble and gravel was returned to the
sampling stations after being picked clean. The benthic organisms
were then preserved in 95% alcohol.
17
Figure 5. --Grid method of selecting one-
square-foot random bottom fauna stations at stream
sampling areas.
18
2| feet
| 3 || |2|| || || |O 9 || 8 || 7 || 6 || 5 || 4 || 3 || 2 || |
9.8 feet | 3.2 feet
|7.5 feet
Figure 5


19
Periphyton Sampling
A study of the effects of TFM on periphyton growth was
made during the summer and fall of 1964 and 1965. Twelve plexiglass
plates 2 inches by 5 inches were installed at each periphyton area.
The plates were numbered from 1 to 12 with plate No. 1 on the west
side of the stream and plate No. 12 on the east. None of the plates
were placed in slack water and efforts were made to assure
uniformity between experimental and control areas by selecting
areas with comparable stream velocity and light intensity. The
12 plates for each area were set in a series across the stream.
They were positioned 7 inches below the surface of the water and
parallel to the current direction to eliminate collection of sediment
particles.
The plexiglass substrates were left in the stream for a
period of 14 days to insure the accumulation of a weighable amount
Of periphyton. The sampling period extended from July through
November and included both treatment periods with TFM.
When the substrate plates were picked up, they were
kept out of the sun and the macroscopic Organism S, mainly
blackflies, were carefully picked from the plates. The plates
were then packed in individual freezer bags and kept on ice until
returned to the laboratory. Fresh plates were installed at each
station immediately after the samples were collected.
20
The periphyton plates were immediately placed in a
refrigerator when returned to the laboratory. The growth was
scraped from the top and bottom of each plate, using a microscope
slide and a rubber policeman. The substrate was rinsed with
filtered water and the sample collected in individual 2-ounce bottles.
Before resetting in the stream, the plates were rinsed in 0.01 N HCl
followed by a rinse in distilled water.
IMillipore filter papers, with a disc diameter of 0.47 mm,
were used to concentrate and weigh the periphyton samples. The
experimental error in weighing periphyton papers was determined
by the following procedure: Papers were weighed on a balance,
filtered with distilled water, dried in a dessicator for 48 hours, and
then reweighed. The average error from five weighing procedures
was found to be 0.0006 g.
Each millipore filter paper was weighed on the balance and
stored in the dessicator until used. The periphyton sample was then
filtered through one of the weighed papers using a vacuum filter.
The filter paper and periphyton sample were placed in a dessicator
and dried for 48 hours. When the drying period was over, the papers
were again weighed and the difference gave the dry weight of the
periphyton sample.
When a weighable amount of periphyton was not present on
a single substrate, four plates were pooled into one sample. In these
instances, the corresponding plates in both experimental and control
areas were pooled.
RESULTS AND DISCUSSION
It has been shown in several studies that the action of the
larvicide is dependent upon physical and chemical conditions of the
treated water. According to Applegate et al. (1961) the toxicity of
TFM is strongly influenced by alkalinity and pH, but only slightly
by temperature and oxygen. The chemical is most effective in
killing lamprey larvae in soft, acid waters. Considerably higher
concentrations are required as pH, conductivity and alkalinity
increase. However, the differential toxicity of TFM to lampreys
and other fishes appears to be maintained regardless of the chemical
conditions and concentrations Of TFM encountered.
Some of the physical and chemical properties of the East
Branch of the Chocolay River on September 20, 1965, prior to the
first treatment with TFM are given in Table 1. These conditions
of pH, conductivity and alkalinity fall near the middle of the range
for streams in the state of Michigan. There was a total alkalinity
of 68.0 ppm calcium carbonate and a pH of 7.75. According to
Applegate et al. (1961) for streams with approximately this
alkalinity and pH, a minimum lethal dose of TFM would be 2 ppm
and the maximum allowable dose would be 8–9 ppm. Prediction of
21
22
Table 1. --Some physical and chemical properties of the water
of the East Branch of the Chocolay River on September 20, 1965,
at the TFM treatment site
Aluminum
Calcium
Chloride
Copper
Alkalinity
Phenolphathalein
Methyl purple
Total
IrOn
Nitrogen
Nitrate
Nitrite
pH
Conductivity
Water temperature
0.13 ppm
44.00 ppm Calcium carbonate
2.00 ppm
0.04 ppm
0.00 ppm Calcium carbonate
42.00 ppm Calcium carbonate
68.00 ppm Calcium carbonate
0.30 ppm
1.00 ppm
0.00 ppm
7.75
116 pu ohm
5.1 F
23
the precise toxicity for lampreys and fish in the East Branch could
only be obtained through bioassay techniques.
Water stage, temperature and current velocity data for the
study section of the East Branch during the sampling period in 1965
are given in Table 2. The stage recordings and current velocity
measurements indicate relatively stable water levels with only
moderate fluctuations from the end of July to the middle of October.
The conditions observed during the treatment study indicate that
fluctuations of water temperature and velocity were probably not
abnormal. However, the data collected on October 21 show a sharp
increase in flow over the previous two months' average. This
change came several days after the second treatment with TFM
and after the bottom fauna sampling was completed.
The maximum and minimum temperature data show wide
daily variation. This was probably due to the lack of cover upstream
from the study section. According to Applegate et al. (1961), the
use of TFM as a lampricide is not impaired by low water temperatures;
in fact, the differential toxicities are improved slightly. As the
temperature is lowered from 55 F to 35 F, trout mortalities are
reduced slightly, whereas lamprey mortalities remain essentially
the same.
The water temperatures of the East Branch during both
treatment operations are given in Table 4. On September 24, 1965,
the temperature remained constant at 47 F. During the second
24
Table 2. --Temperature, water stage and current velocity for the study
section of the East Branch of the Chocolay River in 1965
tºur Water Current velocity at bottom
Date Mini- Maxi- Stage fauna stations (ft/sec)
(feet) A B C D
Iſl U11 Yl Iſll II Yl
July 28 37 74 1. 3 0. 7 0. 9 0.2 0.2
July 31 56 64 1. 0 0.8 0.8 0.2 0.1
Aug. 13 50 70 0. 9 0.9 0. 7 0. 1 0. 2
Aug. 31 50 66 1. 6 1. 6 1.4 0.4 0. 9
Sept. 18 ° 44 59 1. 8 1. 8 1. 6 0. 7 1. 1
Oct. 2 37 54 1. 9 1. 9 1. 5 0.8 1.2
Oct. 14 ° 37 50 1.2 1. 1 0.9 0.3 0.4
Oct. 21 38 54 2.2 2. 3 3.4 1.4 1. 6
* First treatment with TFM on September 24, 1965.
b Second treatment with TFM on October 15, 1965.
25
treatment the stream water averaged 43 F. It is felt that the
temperatures encountered during this study period would not have
altered the action of TFM to any extent.
Bioassays
The stream treatments with TFM were made on September 24
and October 15, 1965. Since the amount of TFM needed for lamprey
control could not be determined from chemical analyses of the water,
each treatment was preceded by a bioassay. Physical and chemical
properties of the water, plus the biological activity Of TFM as
determined through these pre-treatment bioassays, are given in
Table 3. The first bioassay, performed on September 21, gave a
minimum lethal dose of 1.0 ppm TFM and a maximum allowable
dose of 4.0 ppm.
The first treatment was made at the concentration of 1.0 ppm
TFM, which wº the minimum lethal dose. The brook lampreys
that were held in the stream were not killed within the ensuing
24-hour period. Apparently the 1 ppm concentration was not
suitable and plans were immediately made for a second treatment
at a higher concentration.
The second treatment On October 15 was made using a
concentration of 4.0 ppm or 1 ppm below the maximum allowable
dose. The test lampreys in the stream were killed quickly at this
concentration. Water temperatures and the variation in concentration
26
Table 3. --Some physical and chemical properties of the East Branch
of the Chocolay River, and the biological activity of the larvicide in
this water as determined from pre-treatment bioassays
Test Minimum Maximum Conduc – is tº
e - Alkalinity
Date, tempera— lethal allowable tivity hth MO H
1965 sure dose TFM dose TEM at 20 g § m) (ppm) p
(°F) (ppm) (ppm) (unho) \PPm) (PP
Sept. 21 55 1. 0 4. 0 117 0 43 7.75
Oct. 14 52 2.0 5. 0 122 0 48 tº-
1 Concentration of TFM killing 100% of the test lamprey larvae within
24 hours.
2 Concentration of TFM killing approximately 25% of the test rainbow
trout within 24 hours.
27
of TFM used in both treatments are given in Table 4. The desired
concentration of 1.0 ppm was maintained very closely during the
first treatment; however, for the second treatment, the actual
concentration fluctuated considerably from the desired 4.0 ppm.
BOttom Fauna
Bottom fauna areas were sampled on a schedule designed
to detect any immediate effects of TFM on the benthic community.
The experimental and control bottom fauna areas were sampled
and compared to discern changes in abundance. Significant
mortalities, due to the lampricide, should been detected by this
analysis. Long-term or sub-lethal effects of TFM upon the
invertebrates would not have been detected. Torblaa (1968) found
that aquatic invertebrate abundances were not significantly
different one year after treatment with lampricide, suggesting that
there is very little, if any, long-term effect upon the stream
invertebrate communities. Smith (1967) conducted laboratory
bioassays to determine the effects Of TFM on aquatic invertebrates.
He found that it is potentially toxic to some of the invertebrates but
that mortalities would probably be low at concentrations used for
In Ost stream treatments.
The sampling and treatment schedule for the riffle and
pool areas respectively are given in Figures 6 and 7. Benthic
samples from One area on One day (10-square foot samples) are
28
Table 4. --TFM concentrations with variations and
temperature in the East Branch of the Chocolay
River during treatments
Variation from
Termpera- e
º p TFM desired
Time ture º
O (ppm) C Oncentration
(9 F) -
(ppm)
First treatment September 24, 1965
0900 47 1.0 0. 0
1000 47 1.2 +0. 2
1 100 47 1.2 +0. 2
1200 47 1. 1 +0. 1
1300 47 1.0 0.0
1400 47 1.2 +0. 2
Second treatment October 15, 1965
0.855 43 4. 1 +0. 1
09 10 43 4.8 +0, 8
09:50 43 3. 3 –0. 7
1 100 - 43 6. 1 +2. 1
1145 43 7.2 +3. 2
1230 43 6. 2 +2.2
1300 44 3. 8 –0. 2
1330 44 3. 9 –0. 1
1400 44 5. 0 +1. 0
29
Figure 6. --Temporal arrangement of riffle
bottom fauna sampling schedule with mean number of
benthic organisms per square foot in parentheses.
30
Riffle
- (194.4) (122. 6) (104.3)
Control IB1—IB2 y 3
Experimental IA1 – IA2 — IIA1 IIA2
(164.8) (164.0) (161. 6) (107.4)
Date 9–14 9 – 29 10-13 10- 18
First treatment Second treatment
9–24 10–15
Area B – Control riffle above treated section
Area A - Experimental riffle in treated section
I - Means associated with first treatment
II - Means associated with second treatment
1 – Means before treatment
2 - Means after treatment
3 - Means during treatment
Lines connecting areas represent statistical comparisons
by three-way analysis of variance
Figure 6
31
Figure 7. --Temporal arrangement of pool
bottom fauna sampling schedule with mean number of
benthic Organisms per square foot in parentheses.
32
POO1 (46.8) (21.6) (26.4)
Control ID 1 — ID2 IID3
Experimental IC 1 — IC2 — IIC 1 IIC2
(30.9) (26. 3) (47.0) (23.6)
£
Date 9 – 16 t 9 – 28 10 – 14 10 – 18
- First treatment Second treatment
9 – 24 10 – 15
Area D – Control pool above treated section
Area C – Experimental pool in treated section
I - Means associated with first treatment
II - Means associated with second treatment
- Means before treatment
Means after treatment
- Means during treatment
:
-
Lines connecting areas indicate comparison by
three-way analysis of variance
Figure 7
33
characterized by three symbols. The first, a roman numeral,
indicates which treatment period. The second, a capital letter,
tells which area was sampled; and third, an arabic numeral, tells
whether the time of sampling was before or after the treatment.
Before and after samples were collected for both treatments
except for control areas (B and D) during the second treatment.
Because of a shortage of labor, these two areas were only sampled
once; at the time of treatment. It was hoped that this would be
sufficient for making meaningful comparisons.
Seven sets of samples were taken for both the riffle and
pool areas. For these seven sample groups, nine statistical
comparisons were made for the riffle areas and nine for the pool
a £6 a S.
The bottom fauna samples were sorted and identified to
genera for most of the benthic invertebrates utilized in the analyses.
The taxonomic descriptions and keys used for identification were
those provided by Burks (1953), Frison (1935), Leonard and Leonard
(1962), Needham and Needham (1962), and Ward and Whipple (1918).
A qualitative list of all macrobenthic Organisms collected during
this study is given in Table 5. The bulk of the organisms in the
samples come from the orders Diptera and Ephemeroptera with
the orders Plecoptera and Trichoptera in much lesser abundance.
Six orders, 15 families and 18 genera were represented in the
bottom fauna samples. All instars of each taxon of the benthic
Organisms were lumped together in the counts.
34
Table 5. --List of macrobenthos taxa in the East Branch of the
Chocolay River in the fall of 1965
Order Family Genera
Ephemeroptera Heptageniidae Epe Orus
Stenonema
Baetidae Paraleptophlebia
Ephemerellidae Ephemerella
Plecoptera Perlidae Acroneuria
Perlodidae 2
Taeniopterygidae Taeniopteryx
Trichoptera Rhyacophilidae Glossosoma
Rhyacophila
Hydropsychidae Hydropsyche
Diptera Rhagionidae Atherix
Tipulidae Antocha
Simuliidae 2
Ceratopogonidae Palpomyia
Tendipedidae 2
Coleoptera Elmidae Ancyronyx
2 2
Hydracarina
35
Some of the taxa were not abundant enough for meaningful
comparisons and evaluations, so Only 13 of the 18 taxa were utilized
in this study for testing the effects of TFM. The average number of
individuals per square foot before treatment for each of the taxa of
riffle and pool bottom fauna, selected for evaluation by virtue of their
relatively high abundance and wide variety, are given in Table 6.
They were thought to effectively encompass the range of types of
stream invertebrates which might be adversely affected by the
chemical treatments.
All mayfly genera present in the samples were included
in the analysis because many studies have shown that, in general,
the Ephemeroptera are intolerant to most types o: "pollutants" and
chemicals. Applegate et al. (1961) and others suggested a low
tolerance of this order to TFM. Moyle and Luckman (1964) showed
that mayflies as a group are killed immediately by insecticides and
that populations remain depleted for several years. Surber (1953)
indicated that the mayflies were the least tolerant and first to
disappear under polluted conditions. The Ephemeroptera, because
Of their low tolerance to environmental contamination, should be
among the first taxa to exhibit any deleterious effects of the
lampricide.
Twelve taxa of benthic Organisms were selected from
the riffle fauna for evaluating the lampricide treatments and eight
taxa were selected from the pool fauna. The riffle fauna was more
36
Table 6. --Genera and relative abundance of macrobenthos
selected for statistical comparison from before treatment
samples
Area Taxa º
POOl Stenonema 10. 0
Paraleptophlebia 1. 0
Ephemerella 1. 5
Perlodidae 1. 7
Taeniopteryx 0.4
Hydropsyche 1.5
Antocha 4. 5
Tendipedidae 15. 3
Riffle Epe Orus 22.9
Paraleptophlebia 11. 1
Baetis 9. 6
Ephemerella 6.4
Perlodidae 3. 7
Taeniopteryx 3.4
Glossosoma 6. 0
Hydropsyche 73. 4
Atherix 5.9
Antocha 9. 7
Tendipedidae 22. 1
Hydracarina 5. 5
37
diverse and supported considerably larger numbers of individuals
of those taxa found in both the riffle and pool areas. Seven out of
the eight taxa selected from the pool areas were also utilized in
the riffle analysis.
The mayfly, Epeorus, the net spinning caddis fly,
Hydropsyche, and the midges, Tendipedidae, were the most
abundant members of the benthic community found at the riffle
stations. The mayfly, Stenonema, and the midges were the most
abundant members of the benthos at the pool stations. The
remainder of the thirteen taxa were found to be in relatively
low abundance.
Riffle and pool stations were analyzed separately to
eliminate a large source of variation, even though the benthic
organisms used in the pool areas were also used for the riffle
areas. Differential effects of TFM were anticipated due to
differences in water velocity, depth, and substrate type. These
physical differences and the differences in distribution and
abundance of the taxa between riffle and pool areas necessitated
the separate evaluation.
The samples collected in experimental areas before
treatments were utilized as indices of abundance for comparisons
with post-treatment samples. Pre- and post-treatment samples from
control areas were analyzed for random changes in abundance Or
those beyond the treatment effect. For general comparisons between
taxa, the change in abundance over the specified time interval was
38
computed as the percentage of the former level of abundance. Values
below 100% indicate a drop in abundance and those over 100% an increase
in abundance. These calculations were made to eliminate some of the
variation between benthos abundances and to put the data on more
comparable terms. The changes for each taxon of benthic Organisms
used in the analysis of the riffle areas are given in Table 7. Relative
abundance before the first treatment is given as the mean number of
individuals per square foot (Table 6).
For the first treatment period (IA1–IA2), eight taxa in the
experimental area showed a drop in abundance while four taxa
increased. In the control area, ten taxa decreased during the same
time interval. Only two, Hydropsyche and Antocha, decreased in
the experimental area and not in the control. For the most part,
decreases in abundance were greater in the control area than in
the treated, experimental area. Since the decreases in benthic
abundance were greater in the control area, there was probably no
change in the experimental area due to the TFM.
Changes in abundance for eachtaxon over the 14-day period
from after the first treatment to before the second treatment are
included in Table 7. There appears to have been a slight increase
in numbers in the experimental area although they may well be
within sampling variation.
For the second treatment period (IIA 1-IIA2) ten taxa
showed a decrease in abundance and Only one increased, while
39
Table 7. --Changes in abundance for each genera of bottom fauna from
riffles over the specified time interval calculated as the percent of the
former abundance
Experimental area Control area
Taxa LA1– IA2– IIA 1- IB1 - 1B2–
IA2 IIA1 ILA2 IB2 IIB3
Epe Orus 107 72 102 86 79
Paraleptophlebia 94 138 71 31 152
Baetis 84 96 71. * 34 194
Ephemerella 163 86 48 84 89
Perlodidae 67 103 100 50 106
Taemopteryx 84 185 31 158 195
Glossos Oma 55 221 56 117 106
Hydropsyche 115 70 56 54. 74
Atherix - 113 87 72 74. 100
Antocha 68 127 66 94 49
Tendipedidae 77 161 61 63 112
Pſydracarina 74 109 69 67 31
40
five taxa in the control area decreased in abundance. Since the
benthos in the experimental area decreased considerably in
abundance, more than the benth Os in the control area, the TFM
probably did cause some mortality or movement out of the area.
However the experimental and control sampling times were different.
The control samples were taken on the day of treatment, thus less
value can be placed on their comparison with the experimental.
Each taxon was checked for its reaction through the four
separate treatment periods. Seven riffle taxa declined in abundance
after successive treatments with larvicide. Two of these,
Hydracarina, and the dipteran, Antocha, also declined in the
control area during both sampling periods. Two other genera,
Ephemerella and Hydropsyche, declined during both control
periods, but Only during the second treatment in the experimental
riffle. All taxa declined in abundance during the second riffle
treatment period except for Epeorus and Perlodidae which both
remained at the pre-treatment level.
Smith (1967) found that, under lab conditions, a concentra-
tion of 4 ppm TFM would cause short-term mortalities of less than
25% for each of the groups of invertebrates represented by the
12 taxa. In this study some taxa decreased in abundance more than
25% but most of these showed considerable variations under the
control and untreated conditions.
41
The pool community had a lower density and was more
variable in abundance and had lower species diversity, than the
riffles. The changes in abundance over the specified time interval,
as the percent of the initial abundance, for each of the eight taxa
utilized in the pool comparisons are given in Table 8.
After the first experimental treatment period (IC1-IC2)
there were decreases in the abundances of five taxa and increases
in three. However, all eight taxa decreased in the untreated
area during the same time period. Hence, no harmful effects of
the lampricide could be demonstrated in the pool areas during
the first treatment.
The 16-day period between treatments was characterized
by increases in abundance for all taxa in the experimental area.
The control area showed six taxa increased and two decreased
between treatments. It appears that the taxa in the experimental
pool area were not very different from the control pool at completion
of the first treatment.
The second treatment period (IIC 1-IIC2) resulted in
declines in abundance for all taxa in the experimental area. The
control pool area showed six taxa increased and two decreased.
Here again it appears, in the experimental area, that the treatment
has resulted in substantial drops in the quantity of benthic inverte-
brates. However, when the control samples are compared, the wide
fluctuations in abundance make a judgment nearly impossible.
42
Table 8. --Changes in abundance for each genera of bottom fauna from
pools over the specified time interval calculated as the percent of the
former abundance
Experimental area Control area
Taxa IC 1 – IC2– IIC 1 - ID 1 – TD2-
IC2 IIC 1 IIC2 ID2 IID3
Stenonema 109 108 56 44 144
Paraleptophlebia 62 620 61 - 25 43.3
Ephemerella 36 183 64 7 300
Perlodidae 80 120 92 67 67
Taeniopteryx 567 1 12 21 50 50
Hydropsyche 440 132 62 29 229
Antocha 67 117 48 29 1 11
Tendipedidae 60 295 42 53 133
43
Torblaa (1968) found that most groups of invertebrates
declined in treated streams one week after treatment with TFM.
He also showed that most groups had very rapid recoveries, usually
after one week. Likewise he found a wide variability in numbers of
Organisms.
Statistical comparisons between the experimental and
control areas were made by a three-way analysis of variance with
ten replications. This analysis tested the differences between the
means for the three main effects (sources of variation): area or
treatment, stations or samples, and taxa. Three interactions were
also tested to see if any combination of effects was significant.
Significant differences, at the 95% level, were expected
between the different taxa of bottom fauna because they are
normally different in abundance and distribution. Some variation
was also expected between samples (stations) within each area but
not to a significant degree since sampling procedures were carefully
controlled to reduce sampling error. Significant differences were
expected when testing the treatment effect in the experimental areas,
and non-significant differences in the corresponding control areas
for that same effect. Such a result would show that the lampricide
had caused mortalities or m Overments of the benthic communities
within the treated stream. The control area comparisons would also
reveal any extraneous or natural changes in invertebrate abundances or
natural differences between the experimental and the control areas.
44
The F-values for the six sources of variation for nine
analyses of variance comparing riffle areas and nine analyses
comparing pool areas, respectively, are given in Tables 9 and 10.
The relative importance of these nine comparisons can be better
understood by looking at the mean number of benthic invertebrates
per square foot as shown in Figures 6 and 7. The statistical
comparisons over the time periods were made to analyze the
treatment effects. Those comparing the control and experimental
areas were made first to establish that these areas were
statistically identical and to follow the eventual changes in the
benthic communities more closely.
The 95% level was established as the level of significance
and any F-value equal to or exceeding it was considered significant.
Any general mortality or change in abundance should cause
a significant F-value for the area term. This treatment source of
variation is probably the most meaningful because it compares the
total number of invertebrates between the two areas. The mean
numbers of benthic organisms per square foot were 194. 4 in the
control riffle and 164.8 in the experimental riffle, so the control
riffle had 29.6 more individuals per square foot than the
experimental riffle area. However, the non-significant F for the
area term comparing the experimental and control areas before
treatment (IA1-IB1) establishes a statistical equality in benthic
i.
Table 9. --F-values for sources of variation from three-way analysis of variance comparing riffle
bottom fauna samples 1
Source df LA 1- IB1– IA 1- IA2– IB2– IA2– IIA 1- IIA 1- ILA2–
oure IB1 IB2 IA2 IB2 IIB3 IIA 1 IIB3 IIA2 IIB3
Area 1 0.76 3. 32 0.002 2. 36 0. 57 0.02 11. 21% 11. 65% 0.05
Stations 9 1. 62 0.91 1. 54 0.82 0. 90 5. 37% 2. 73% 3.25% 1. 71
Taxa 11 15.86%. 13.86% 22.22% 13. 64% 13. 59% 28. 56% 17. 80% 15.17%. 17. 63%
Area x
station 9 1. 71 1. 30 3.01% 1. 35 1. 22 2. 35% 2.93% 1. 91 1. 44
Area x
taxa 11 3. 19% 1. 30 0.03 0. 73 0.46 2. 07 1. 59 1. 10 3. 86%
Station X
taxa 99 0.97 0. 70 0. 90 0. 54 0.68 1. 95 1. 17 0.83 0. 70
Error 99
Significant F-values at the 0.95 level or above are indicated by *.
§
Table 10. --F-values for sources of variation from three-way analysis of variance comparing pool
bottom fauna samples Nº.
SOurce df IC 1 – ID 1 – IC 1– IC2- ID2– IC2– IIC 1– IIC 1 - IIC2-
ID1 & ID2N22 IC2 TD2 IID3 IIC 1 IID3 IIC2 IID3
Area 1 4. 27%; 20.24% 1.42 3.99% 4. 25% 17. 0.1% 20.67%. 23. 25% 0.49
Stations 9 1.42 1. 13 2.67% 1. 18 1. 37 0.62 2. 03 0.99 2. 23%
Taxa 7 31. 90%. 28.88% 32.86% 28. 56% 4.1. 15% 35. 14% 55. 19%. 38.90% 29. 1.1%
Area x
station 9 0.96 1.49 1. 48 1. 95 2.72% 1. 75 1. 36 1. 40 1. 23
Area x
taxa 7 1. 41 3. 25% 2. 53% 2. 03 1. 0.8 8. 44% 4. 17%. 65.95% 0.95
Station x
taxa 63 0.93 0.94 0.97 0. 73 1. 36 0. 71 1.42 0.86 1. 10
Err Or 63
* Significant F-values at the 0.95 level or above are indicated by *.
& Degrees of freedom are different for IC 1-ID1 and ID 1-ID2 because of a missing station.
47
abundances between the two areas. Sampling error is probably
high enough to mask any difference in abundance between control
and experimental area.
Two of the other eight F-values for the treatment term in
the riffle area were significant and indicated that the benthic
abundances were different in those areas. One occurred during
the second treatment in the experimental area and the other resulted
from a comparison between the experimental riffle area before the
second treatment and the control riffle during the second treatment.
The significant term for the experimental area during the
second treatment (IIA 1-IIA2) is important because it shows that a
change in abundances had taken place, while a non-significant F in
the control area (IB2–IIB3) would establish the treatment with
lampricide as the cause. A problem is encountered because there is
a difference in the time of sampling the control area (B) and
experimental area (A) during and after the second treatment.
Since the control area was sampled on the day Of the second
treatment, the result in (IB2–IIB3) is not as easily compared.
The non-significant F for the (ILA2–IIB3) area term is also
confusing because it implies that the experimental benthic
community had not changed from that of the control. This
situation can probably be explained by the mean benthic numbers
given in Figure 6. The control area, which had not changed
significantly between sampling periods, had continually been
48
decreasing in mean number of individuals per square foot up to the
last sampling period. A three-way analysis of variance comparing
the control riffle before the first treatment (IB1) and during the
second treatment (IIB3) was significantly different at the .995 level
(F = 9.7 ) validating the drastic and regular decline in the
*1, 99
control benthic numbers.
The mean number of Organisms per square foot in the
experimental riffle area remained nearly constant until the second
treatment. The decrease in the experimental riffle of 54.2
individuals per square foot after the second treatment was highly
significant and when compared with the extremely small changes in
that area for the previous sampling periods, gives strong indication
that the second treatment with TFM (4.0 ppm) Caused a significant
decline in the riffle benthic abundances.
The significant F-values for the station term show that
enough variation was present to make the stations different. This
was not necessarily the reason for a significant area term because
(IA2–IIA1) had a significant station term and a non-significant area
term.
Even though the larvicide probably caused a decrease in the
benthos, the conditions were satisfactory for rapid recolonization.
None of the taxa were eliminated entirely and substantial populations
probably were present upstream from the study area which could
provide enough drift to compensate for a small effect. Waters (1964)
49
found that the mayfly, Baetis, returned to 100% of its former
density 4 days after removal. He also has shown that drift is a
mechanism capable of returning disturbed populations of many
stream invertebrates to normal or capacity levels in a relatively
short time.
The wide variation in the pool benthos makes analysis of
the information very difficult. The significant F-value for the area
term comparing the experimental and control pool areas before
treatment establishes that these invertebrate communities were
statistically different (Table 10). Also nearly all of the treatment
terms were significant for both the control and experimental areas.
The mean number of Organisms per square foot dropped in both the
experimental and the control pool area during the first treatment
period. Both areas then increased to the second treatment period.
There was a very substantial decrease of 23.4 benthic organisms
per square foot in the experimental pool area during the second
treatment. This decrease indicates that the second treatment at
4 ppm TFM did significantly lower the benthic abundance in the
experimental pool area.
The control pool was also highly variable and the
non-significant area term between the experimental after the
second treatment (IIC2) versus the control during the second
treatment (IID3) is probably a circumstance of that variation. The
control pool area certainly did not provide a good measure for the
50
effects of the TFM in the experimental pool area, and little value
can be placed upon these comparisons in judging the effects of the
lampricide upon the benthic fauna.
Periphyton Results
Periphyton growth was of interest in this study because
it provides the basis of the stream's productivity and because little
work has been done on its reactions to environmental changes.
Periphyton growth was examined during the years 1964 and 1965.
This community Of Organisms is made up of those that are
attached or move upon submerged substrates. Reid (1961) says
that the periphyton, typically, is an assortment of unicellular
and filamentous algae with various attached protozoans, bryozoans
and rotifers. Sladeckova and Sladecek (1962) define the true
periphyton as those organisms which are attached, thus immobile,
and which show various adaptations for sessile life. Sladeckova
(1962a) found in a new reservoir that this group contained bacteria,
algae, fungi, and rotifers. Clifford (1959) found that the periphyton
community on artificial substrates in a Michigan stream was
composed almost entirely of diatoms.
Stream velocity is thought to be an important factor in
periphyton growth. It was measured at each area on the days of
sampling as shown in Table 11. The importance of current
velocity was made evident by Whitford (1960) when he demonstrated
51
Table 11. --Current velocities at periphyton areas in feet per
second. (Each value is an average of four measurements.)
Date Area Area Area Area
Y Z X W
1964
Aug. 13 1. 1 1.2 0.8 0. 9
Aug. 31 1.4 1. 7 0.6 0.8
Sept. 14 1.8 1. 9 1. 1 1. 1
Oct. 12 0.3 0. 1 0. 1 0.3
oct. 26 2. 6 3. 8 0. 7 1. 6
NOV. 9 3. 9 3. 8 0. 9 1. 8
1965
June 23 1.2 1. 1 G- 0.6
July 7 0.8 1. 1 0. 1 0.4
July 21 0.8 0. 7 0.2 0.3
Aug. 4 0.6 1. 3 0.2 0.4
Aug. 18 0. 9 0.8 tº- -
Sept. 1 1. 7 1. 8 0. 5 1. 7
52
that many species of attached algae grew best in a current and that
some died when placed in still water. Kevern and Ball (1965) also
demonstrated higher periphyton productions with higher current
velocities.
Periphyton standing crop in the Chocolay River was
measured by allowing growth of the communities on the submerged
plexiglass plates for a 14-day period. Sladeckova (1962b) used this
method because the quantitative removal of the periphyton is easily
accomplished.
The periphyton was collected, dried and weighed to measure
the standing crop. The weight of Organisms on a suitable, uniform
surface is a more accurate and direct measure of the productive
capacity of waters than other techniques (Cooke, 1956).
Figures 8 and 9 contain graphs of periphyton dry weights
Over the 2-year sampling period. The graph in Figure 8 shows the
two pool sampling areas (W and X). Standing crop in these two
areas remained very consistent Over the study period. The control
pool produced higher weights than the experimental pool area for
most of that time. It is quite interesting to note that standing crop
in both years showed three distinct peaks on approximately the
same dates. It is also of note that area (X) had consistently slower
current velocities than did area (W), and yet, still exhibited higher
standing crops. It is possible that the increases in periphyton
standing crop with increased current velocity occurred but were
53
Figure 8. --Graph of periphyton dry weights over
the entire study period for the pool areas.
54

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56

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8/13 8/3| 9/14 9/28 IO/I2 IO/26 II/9
1964
Figure 9
57
not measured because of a high rate of sloughing and scouring which
removed substantial amounts of periphyton from the plates. Whitford
(1960) indicated that current velocities must exceed 0.5 feet per
second to produce the steep diffusion gradient that is beneficial to
growth. Kevern and Ball (1965) showed increased production with
much lower increases in current velocity.
The graph of the periphyton weights in the pool areas does
not show any drastic effects from the treatments with lampricide.
Growth in both areas rose to a peak after the second treatment and
then dropped off sharply in mid-November, as it had done in 1964.
The graph in Figure 9 shows the periphyton standing crops
in riffle areas (Y and Z). The weights for these areas were
considerably lower than those found in the pool areas. Again the
explanation might be that the current velocities were high and might
have sloughed off enough periphyton to allow greater weights in the
pool area, not taking into consideration. Other unmeasured factors
such as light and Oxygen tensions. McIntire (1966) stated that in
laboratory tests run at high Oxygen tensions, rates of growth were
higher for the slow current periphyton communities than for the
fast current communities.
The experimental riffle area (Y) and the control riffle
area (Z) produced nearly equal weights of periphyton at each time
of sampling and they fluctuated together. Here, as with the pool
58
areas, the treatments did not appear to affect the periphyton growth
which was actually higher in the treated area after the second
application of TFM.
Statistical comparisons between the periphyton weights in
the experimental and control areas from 1964 and 1965 were made
by a four-way analysis of variance with two- and three-way
interactions. The F-values for the sources of variation are given
in Table 12. There was a significant F, at the 95% level, for all
four sources of variation. The weights were different between the
two years, they were different on the various dates sampled, and
the weights in pools and riffles were different. The weights in the
experimental areas were also different from those in the control
areas. This variation in standing crops is probably too large to
make meaningful statistical comparisons between the areas.
There is little basis in this study for measuring the effects
of the larvicide upon periphyton standing crop. The dry weights
from both 1964 and 1965 show a drastic decline in October and
November which is probably due to shortened photoperiod and cold
water temperatures. These fluctuations at the time of treatment
tend to mask any effects which might be due to the TFM.
Mean periphyton weights for the riffle and pool areas
before and after each of the treatments with larvicide are given
in Table 13. A two-way analysis of variance with 1 and 5 degrees
of freedom with Orthogonal contrasts was used to test whether the
59
Table 12. --F-values for sources of variation from four-way
analysis of variance comparing periphyton sampling areas
Source df SS MIS F
Total 53 . 0046111
Years 1 - 000 12025 - 000 12025 8. 95%;
Dates 6 . 00182944 . 00030490 22. 69***
water * 1 - 000 84.846 . 000 84.846 63. 13×2k;
Area 3 1 . 000 18793 . 00018793 13. 98***
Year x dates 6 . 0004 1359 . 00.006893 5. 13%;
Year X. Water 1 .00007731 . 00007731 5. 75%
Year X area. 1 . 00000 706 . 00000 706 0. 53
Date X Water 6 . 00032349 . 00005391 4. 0.1%
Date x area 6 . 00000000 . 00000000 0.00
Water X area 1 - 00014331 - 00014331 10. 66%;
Year x dates x water 6 . 00047.606 . 00007934 5. 90%;
Year X. Water X area 1 . 00000 844 . 00000 844 0.63
6 . 00009 138 . 0000 1523 1. 13
Date x water x area
Err Or 10 . 00013439 . 00001344 -
1 Significant F-values at the 0.95 level are indicated by *,
at the 0.975 level by **, and at the 0.995 level by ***.
Comparison between riffle and pool areas.
3 e tº
Comparison between experimental and control areas.
60
Table 13. --Mean periphyton weights in grams per square meter from
two sampling periods immediately before and after each treatment
With TFM
First treatment Second treatment
period period
Experi- Control Experi- Control
mental mental
RIFFLE AREAS
Before treatment
First mean 1. 44 2.48 . 82 0.85
Second mean 0. 82 0.85 . 19 0.39
After treatment
Third mean 0. 19 0.39 . 62 0.37
Fourth mean 0.62 0.37 . 19 0. 11
POOL AREAS
Before treatment
First mean 2. 68 2. 71 . 53 1. 60
Second mean 0. 53 1. 60 . 65 3. 57
After treatment
Third mean 0.65 3. 57 . 12 4. 17
Fourth mean 4. 12 4. 17 . 77 3. 64
61
mean standing crops were significantly different. Eight analyses
were made comparing the experimental and the control areas during
the two treatment periods. Two means before each treatment were
compared with two means after the treatment for each of the four
study areas. Eight more analyses were run comparing each of the
four experimental areas with each of the control areas to test
whether they were significantly different. None of the resulting 16
F-values were significant at the 0.95 level.
The direction of the changes of the IY) ea.ſ.l. standing crops during
the two treatments also does not indicate any adverse growth changes
due to the lampricide. Only the experimental riffle area during the
first treatment showed a decreased mean weight and the corresponding
control area also decreased. All other periphyton areas showed no
change Or increased Standing crops during the treatment periods. So
there is no evidence in this study that the periphyton standing crops
were affected by either of the larvicide treatments.
SUMIMARY
1. The Chocolay River, Marquette County, Michigan, was
selected to study the effects of lamprey larvicide on the bottom fauna
and periphyton. In 1965, two stream treatments with TFM
(3-trifluormethyl-4-nitrophenol) were made. The concentration of
larvicide was 1 ppm during the first treatment and 4 ppm during the
second. Bottom fauna and periphyton samples were collected before
and after each treatment to analyze the effects of the larvicide.
2. In order to evaluate the effect of the larvicide, twelve
taxa of bottom fauna in riffle areas and eight taxa in pool areas were
utilized. Percent change in abundance of each taxa did not reveal a
larvicide effect from the 1 ppm treatment. However, all taxa, except
two, decreased in numbers in the experimental riffle area from the
4 ppm treatment. The eight taxa in the experimental pool area also
decreased in numbers during the 4 ppm treatment. These results
indicated that the second larvicide treatment did cause a decline in
bottom fauna.
3. Numbers of benthic Organisms in the experimental and
control areas were statistically compared. In the experimental riffle
areas, the number of benthic Organisms was significantly lowered
after the second treatment. No other significant differences for the
62
63
bottom fauna in the riffles were found except between the experimental
and control number before the second treatment. This difference was
probably a function of the high variation in the control.
Pool bottom fauna areas exhibited high variation unrelated
to TFM and most statistical comparisons showed the experimental and
control to be significantly different in numbers of benthos during most
sampling periods. These differences make a judgment on larvicide
effects inpossible for the experimental pool benthos.
The only positive result of statistical analyses strongly
indicated that the 4 ppm larvicide treatment caused a decline in
rine bottom fauna abundance, but not the pool abundance.
4. During 1964 and 1965, periphyton standing crop was
measured at two experimental and two control areas to test effects
of the larvicide. A statistical analysis comparing periphyton weights
revealed that standing crops were significantly different between years,
areas, and within areas on different dates. Periphyton weights immediately
before and after each treatment were compared to further test the
effects of the larvicide treatments. No significant differences were
found suggesting that the larvicide did not affect the standing crop of
periphyton.
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