INVESTIGATIONS IN FISH CONTROL

68. Development and Evaluation of On-site Toxicity Test 

Procedures for Fishery Investigations

United States Department of the Interior

Fish and Wildlife Service
Investigations in Fish Control, published by the Fish and Wildlife Service, include reports on the 

results of work at the Service's Fish Control Laboratories at La Crosse, Wis., and Warm Springs, 

Ga., and reports of other studies related to that work. Though each report is regarded as a separate 

publication, several may be issued under a single cover, for economy. [See Investigations in Fish 

Control 47-50 (in one cover) for list of issues published prior to 1970.]

35. Toxicology of Thiodan in Several Fish and Aquatic Invertebrates, by Richard A. Schoettger. 

1970. 31 pp.

(Reports 36 through 38 are in one cover.)

36. A Method for Rating Chemicals for Potency Against Fish and Other Organisms, by Leif L. 

Marking. 1970. 8 pp.

37. Comparative Toxicity of 29 Nitrosalicylanilides and Related Compounds to Eight Species of 

Fish, by Leif L. Marking and Wayne A. Willford. 1970. 11 pp.

38. Toxicity of 33NCS (3'-chloro-3-nitrosalicylanilide) to Freshwater Fish and Sea Lampreys, by 

Leif L. Marking, Everett L. King, Charles R. Walker, and John H. Howell. 1970. 16 pp.

(Reports 39 and 40 are in one cover.)

39. Effects of Antimycin A on Tissue Respiration of Rainbow Trout and Channel Catfish, by 

Richard A. Schoettger and Gerald E. Svendsen. 1970. 10 pp.

40. A Resume on Field Applications of Antimycin A to Control Fish, by Robert E. Lennon and 

Bernard L. Berger. 1970. 19 pp.

(Reports 41 through 43 are in one cover.)

41. Identification of MS-222 Residues in Selected Fish Tissues by Thin Layer Chromatography, by 

John L. Alien, Charles W. Luhning, and Paul D. Harman. 1970. 7 pp.

42. Dynamics of MS-222 in the Blood and Brain of Freshwater Fishes During Anesthesia, by 

Joseph B. Hunn. 1970. 8 pp.

43. Effect of MS-222 on Electrolyte and Water Content in the Brain of Rainbow Trout, by Wayne A. 

Willford. 1970. 7 pp.

44. A Review of Literature on TFM (3-trifluormethyl-4-nitrophenol) as a Lamprey Larvicide, by 

Rosalie A. Schnick. 1972. 31 pp.

(Reports 45 and 46 are in one cover.)

45. Residues of MS-222 in Northern Pike, Muskellunge, and Walleye, by John L. Alien, Charles W. 

Luhning, and Paul D. Harman. 1972. 8 pp

46. Methods of Estimating the Half-Life of Biological Activity of Toxic Chemicals in Water, by Leif 

L. Marking. 1972. 9 pp.

(Reports 47 through 50 are in one cover.)

47. Preparation and Properties of Quinaldine Sulfate, an Improved Fish Anesthetic, by John L. 

Alien and Joe B. Sills. 1973. 7 pp.

48. Toxicity of Quinaldine Sulfate to Fish, by Leif L. Marking and Verdel K. Dawson. 1973. 8 pp.

49. The Efficacy of Quinaldine Sulfate as an Anesthetic for Freshwater Fish, by Philip A. 

Gilderhus, Bernard L. Berger, Joe B. Sills, and Paul D. Harman. 1973. 9 pp.

50. Residue of Quinaldine in Ten Species of Fish Following Anesthesia with Quinaldine Sulfate, by 

Joe B. Sills, John L. Alien, Paul D. Harman, and Charles W. Luhning. 1973. 9 pp.

(Reports 51 and 52 are in one cover.)

51. Methods for Simultaneous Determination and Identification of MS-222 and Metabolites in Fish 

Tissues, by Charles W. Luhning. 1973. 10 pp.

52. Residues of MS-222, Benzocaine, and Their Metabolites in Striped Bass Following Anesthesia, 

by Charles W. Luhning. 1973. 11 pp.
INVESTIGATIONS IN FISH CONTROL

68. Development and Evaluation of On-site Toxicity Test 

Procedures for Fishery Investigations

u

United States Department of the Interior

Fish and Wildlife Service

INVESTIGATIONS IN FISH CONTROL

68. Development and Evaluation of On-site Toxicity Test 

Procedures for Fishery Investigations

By Ralph M. Burress

United States Department of the Interior

Fish and Wildlife Service 

Washington, B.C. 1975

CONTENTS

Page

Abstract.................................................................... 1

Introduction ................................................................ 1

Materials and Methods...................................................... 2

Experimental Fish ........................................................ 2

Preparation of Stock Solutions ............................................ 2

On-site Toxicity Test Arrangement........................................ 3

Filling Bags .............................................................. 3

Observations ............................................................. 3

Monitoring Water Quality................................................. 3

Efficacy of Polyethylene Bags for Use in On-site Toxicity Test............... 3

Recommendations for Conducting On-site Tests ............................. 4

Selecting Test Concentrations ............................................. 4

Test Animals ............................................................. 5

Loading Rates ............................................................ 5

Duration of Toxicity Test ................................................. 5

Selecting Treatment Concentrations....................................... 5

Literature Cited............................................................. 6

DEVELOPMENT AND EVALUATION OF ON-SITE TOXICITY TEST 

PROCEDURES FOR FISHERY INVESTIGATIONS

by

Ralph M. Burress

Southeastern Fish Control Laboratory 

Warm Springs, Georgia

ABSTRACT

A simple, inexpensive procedure was developed for conducting on-site tests 

(bioassays) of the toxicity of various concentrations of antimycin on target and 

nontarget fishes in waters to be treated. After preliminary laboratory experiments 

with seven types of containers showed that large polyethylene bags were best for use 

in field toxicity tests, year-round field experiments were performed in ponds. 

Measurements of water quality in large bags containing 284 liters of water were 

closely similar to water quality measurements in ponds throughout the test periods. 

The maximum safe loading level for 96-h tests at temperatures above 18.3 C was 

about 852 g of fish per bag, but this amount could be increased in colder water or in 

shorter tests. Final modifications and evaluations of the method were made on the 

basis of treatments of ponds with antimycin in accordance with data derived from 

on-site tests. The procedure described is adaptable for tests of many other chemical 

compounds commonly used by fishery workers.

INTRODUCTION

With the development of more potent and 

more selective fish toxicants, fishery managers 

have an increasing need to determine the 

concentration of toxicant which is most effective for a specific purpose in a body of water. The 

efficacy of chemicals used by fishery workers is 

affected by water quality, physical and 

biological conditions in the aquatic habitat, and 

the relative susceptibility of different kinds and 

sizes of target organisms. The lack of specific 

information on which to base application rates 

has often resulted in inadvertent underdosing, 

deliberate overdosing, and repeated dosing, all 

of which were detrimental to the success of the 

treatments or to the habitats treated.

The on-site toxicity test (bioassay) procedure 

is one of the more promising methods for 

determining both the concentration and the 

length of exposure required to produce the 

desired level of control of target organisms. 

Despite the pronounced need for information of 

this kind, the on-site test method apparently has 

not been widely used; relatively few investigators who have applied toxicants to 

manipulate fish populations in streams have 

reported using fish and water from the streams 

to be treated to conduct preliminary on-site 

tests. The facilities employed have varied from a 

simple arrangement of fish hatchery troughs 

beside a stream (Lennon and Parker 1959) to a
fully equipped mobile laboratory (Howell and 

Marquette 1962). On-site tests conducted in 

lakes before the application of fish toxicants 

have been performed in large wastebaskets 

(Berger et al. 1967), in 91-liter plastic garbage 

cans (Pfeiffer 1968), in open-ended, 208-liter 

barrels that were pushed into the lake bottom a 

few inches (Vaughn et al. 1974), and in 76-liter 

capacity plastic bags that were included in kits 

provided by Ay erst Laboratory Inc., for on-site 

tests of antimycin (G. C. Radonski, personal 

communication).

The utility and economy of using small plastic 

bags as bioassay vessels are well documented. 

Davis and Hardcastle (1959) used 25-liter bags 

enclosed in cardboard cartons to conduct 

herbicide toxicity tests in the laboratory; Falk 

(1972) conducted acute toxicity bioassays of 

pollutants by placing 20-liter plastic bags 

inside larger nylon mesh bags supported by 

rigid aluminum frames situated in shallow lake 

waters. In both these studies, the authors placed 

10 fish in each bag and supplied continuous

aeration to maintain adequate concentrations 

of dissolved oxygen for 48 or 96 h.

Although dissolved oxygen diffuses through 

polyethylene film (Fremling and Evans 1963), 

the rate of diffusion is so slow that, if small 

plastic bags are used as bioassay containers, 

either accessory aeration must be supplied or 

the loading levels (grams of fish per liter of 

water) must be light and tests must be of short 

duration. In the present study, the objective was 

to use more and larger fish in 96-h tests of 

antimycin without employing either aeration 

devices or bulky supports for rigid vessels. 

Consequently, the study centered on determining the feasibility of employing plastic bags 

large enough to contain 284 liters of water and 

on developing simple experimental procedures 

for their use. During the period July 1968 to June 

1970, numerous laboratory and field experiments were conducted at the Southeastern 

Fish Control Laboratory, Warm Springs, 

Georgia and in four nearby ponds. Bioassay 

procedures developed were tested in field trials 

in Illinois and Arkansas in 1972.

MATERIALS AND METHODS

Laboratory experiments included comparisons of the utility, efficacy, cost, and safety 

of seven types of potential on-site test 

vessels: polyethylene wastebaskets, fiber glass 

containers, metal lard cans, aluminum pails, 

stainless steel pails, large glass jars, and 

polyethylene bags. No single container was 

superior to the others in every respect, but 

polyethylene bags appeared to be the most 

useful, primarily because they are readily 

portable and are available in large sizes from 

numerous firms that manufacture plastic film 

or plastic containers. Consequently, large 

polyethylene bags (0.96 m * 1.65 m, made of 

material 0.076 mm [3 mils] thick) were used in 

conducting 19 on-site toxicity tests with antimycin in four ponds during the period April 

1969 to June 1970. The ponds varied in type from 

a clear, infertile pond to a highly eutrophicpond 

polluted with dairy wastes. Loading rates 

ranged from 0.11 to 2.76 g/1, but were less than 1 

g/1 in 13 of the tests. Certain procedures were 

modified in the exploratory stage of the study, 

but the methods outlined here were followed 

consistently thereafter.

Experimental Fish

In most experiments, antimycin was tested 

against bluegills (Lepomis macrochirus}\ in 

others, a few subadult largemouth bass 

(Micropterus salmoides) or channel catfish 

(Ictalurus punctatus) were included. Most of the 

fish were seined from the ponds in which the 

tests were conducted. These fish, usually captured a day or two before the tests, were held in 

uncrowded live-cages in the ponds while they 

recovered from the stress of collection, and only 

vigorous specimens were selected for testing. 

The few fish from the laboratory that were used 

in field tests also were held at least 24 h in livecages for acclimation to pond conditions. If the 

supply offish was adequate, 10 fish were placed 

in each vessel except when the use of large fish 

made it necessary to reduce the number to avoid 

exceeding the loading capacity.

Preparation of Stock Solutions

Stock solutions were prepared by using a 1-ml 

glass pipette graduated in hundredths to
measure 1.42 ml of Ayerst's undiluted 20% stock 

solution of antimycin [Fintrol-Concentrate(R)] 

into a 1-liter volumetric flask and adding 

acetone to bring the volume up to 1 liter. The 

addition of 1 ml of this stock solution to 284 liters 

of water yielded a l-fJig/l (1-ppb) concentration 

of antimycin (active ingredient), a procedure 

which simplified operations and reduced the 

likelihood of error. The small amounts of 

toxicant needed for determining the concentrations required for selective kills were also 

measured with a 1-ml glass pipette.

On-site Toxicity Test Arrangement

In selecting the site, the crew avoided areas 

where rocks, roots, or other objects might 

puncture the bags. Where no protective 

enclosure for the bags was needed, two steel 

fence posts were driven into the bottom of the 

pond about 7 m apart in waist-deep water, a rope 

was stretched between the posts, and the bags 

were suspended from the rope with stout twine 

(Fig. 1). When necessary, bags were protected 

from sources of mechanical damage (e.g., 

turtles, boats, and flotsam) by making a simple 

exclosure of netting material supported by four 

steel fence posts. The ropes used to support the 

bags were tied diagonally across the exclosure, 

providing support for a cover made of netting 

and eliminating bird depredations that 

sometimes occurred when treated fish surfaced 

in distress.

Filling Bags

In the laboratory, 284 liters of water were 

pumped into a bag and the water level was 

marked. The bag was then emptied, and about 

50 more bags were marked at the same level for 

use in the field. Two men standing in waist-deep 

water were able to fill and handle bags without

difficulty. Water of this depth permits crewmen 

in chest waders to handle full bags without 

dragging them over the pond bottom. The 

preferred method for filling bags involved the 

use of a large plastic waste can from which the 

bottom had been removed. The can, when fully 

inserted into the mouth of the bag, held the bag 

open and gave good support as the bag was 

filled. When water reached the desired level, the 

can was removed, the bag neck was twisted shut 

and secured with a stout rubber band, and the 

bag was suspended from the rope with twine. 

After all bags were filled, each was reopened; the 

toxicant was added and mixed into the water 

thoroughly with a dip net; fish were added; the 

bag was resealed, returned to its place, and 

numbered. In each experiment, two controls 

(untreated) were used; one contained fish and 

the other only water.

Observations

In clear ponds dead fish could be seen at the 

bottom of the bags and removed easily with a 

dip net when observations were made. If the 

water was not clear enough to permit direct 

observation, all fish were dipped out into a 

container. Live fish then were returned to the 

bags; dead fish were discarded. Use of a 

rectangular dip net expedited removal of dead 

fish from the corners of bags.

.Monitoring water quality

Most tests were conducted for 96 h. 

Measurements of pH and dissolved oxygen in 

the pond, in the control bag containing fish, and 

in the control bag without fish, generally were 

made in early morning and late afternoon. A 

two-man crew could set up and conduct a test 

without undue difficulty if work schedules were 

efficiently arranged.

EFFICACY OF POLYETHYLENE BAGS FOR USE IN 

ON-SITE TOXICITY TESTS

The results of the brief initial experiments 

with large polyethylene bags were encouraging. 

Bags of 0.076-mm wall thickness proved to be 

sufficiently strong. Furthermore, struggling 

fish in their death throes invariably retracted 

their fins when they came in contact with the 

bag wall and thus did not puncture the bags. If

seams leaked or the bag was punctured, the 

medium was not diluted because the water flow 

always was outward.

Results of routine static tests conducted to 

compare the toxicity of antimycin to fingerling 

bluegills in glass jars and in plastic bags were 

closely comparable; this indicates that no
appreciable loss of toxicant resulted from 

absorption, adsorption, or reaction with the 

plastic. Before pond tests were begun, other tests 

were performed in outdoor plastic pools containing phytoplankton. Water temperatures in 

the bags were virtually identical with those at 

corresponding depths outside the bags, and the 

pH of water in the bags corresponded more 

closely with that in the surrounding water than 

did the pH in other types of containers. Concentrations of dissolved oxygen remained higher in 

clear bags than in opaque bags, presumably 

because photosynthesis continued at a higher 

rate in clear bags.

The influence of 2-, 3-, 4-, and 5-g/l loading 

rates on pH and dissolved oxygen concentrations were determined in a summer field test. 

When surface temperatures were high (28.0-31.9 

C), the 3-g/l loading with bluegills of intermediate size was the highest that could be 

used for a 96-h test. A 4-g/l load could have been 

used for as long as 48 h, but a 5-g/l load was too 

great for even a 24-h test.

Comparing pH values and dissolved oxygen 

concentrations in bags and various types of 

ponds during different seasons was a primary 

concern during the development and evaluation 

of toxicity test procedures. Water quality data 

collected during the 19 field tests indicated that 

the highest and lowest pH values measured in 

the control bags containing fish were neither 

consistently higher nor lower than those 

measured at the pond surface (Table 1). Thus, 

test results were not biased by differences in pH

attributable to the bags. Comparisons of dissolved oxygen concentrations showed that 

average readings in the control bags containing 

fish tended to be slightly lower than those in the 

pond, whereas the readings in the control bags 

without fish generally were somewhat higher. 

However, no test results were invalidated 

because of oxygen depletion in the bags. If bags 

made from different grades of raw materials 

have different physical and chemical 

characteristics, oxygen may diffuse less readily 

through some bags than others. Preliminary 

tests with each new lot of bags would help 

determine safe loading rates.

After the 19 field trials were completed and 

basic test methods were established, on-site 

toxicity tests were conducted in one pond in 

Illinois and four in Arkansas. The antimycin 

treatments were successful, and were reported 

by Gumming and Gilderhus (in press) and by 

Gumming, Burress, and Gilderhus (1975). 

Properly secured bags withstood heavy rains, 

winds, and even strong currents induced by 

outboard motor operation. There is good reason 

to believe that such tests also could be conducted 

in streams with a slow or moderate current, if 

bags were protected from floating objects. 

Because of the essentially neutral buoyancy of 

the bags, it is possible to anchor them and 

conduct tests below the surface. G. C. Radonski 

(personal communication) stated that he has 

conducted toxicity tests in plastic bags under ice 

and in the hypolimnion at depths as great as 

9.1 m.

RECOMMENDATIONS FOR CONDUCTING ON-SITE TESTS

The methods outlined above provide the basic 

guidelines for conducting an on-site toxicity 

test. The first day's preparations include securing the experimental fish; setting up the posts, 

ropes, and protective netting; and determining 

pH and other water quality factors. On the day 

of the test, the bags are filled, the toxicant is 

added and thoroughly mixed, and the fish are 

added in rapid sequence at the time of day when 

the full-scale treatment will be applied. This 

ensures that water quality conditions during the 

test and the treatment will be as similar as 

possible.

Selecting Test Concentrations

The range of toxicant concentrations to be 

tested depends largely on the size and relative 

susceptibility of the target fishes, the amount of 

population reduction desired, and the water 

quality characteristics (particularly temperature and pH). Not less than five test concentrations should be used, and a sixth bag, with 

fish, should be used as a control. The following 

concentrations of antimycin (A«g/l) are 

suggested when total kills of susceptible species 

are desired: 2.5, 4.0, 5.5, 7.0, and 8.5. If the
target fishes include resistant species such as 

goldfish (Carassius auratus), gar (Lepisosteus 

spp.), or bowfin (Amia calva), concentrations of 

12,15, 18, 21, and 24/ug/l are more appropriate. 

Less antimycin is required when the temperature is above 15.6 C or the pH is below 8.5.

Selective reduction of target species by complete treatment of the aquatic system requires 

much lower concentrations of antimycin. When 

the temperature exceeds 15.6 C and there are 

marked diurnal fluctuations in pH, the following test concentrations of antimycin (jug/1) are 

suggested: 0.20, 0.35, 0.50, 0.75, and 1.0. If pH 

levels are so high that the 1.0-/^g/l concentration is not adequate, the range of concentrations 

should be shifted upward or the treatment 

postponed until water conditions become more 

favorable. The following concentrations (/Ltg/1) 

are more appropriate for use at temperatures 

lower than 15.6 C: 0.4, 0.8,1.2,1.6, and 2.0. The 

duration of toxicity tests should be extended by 

1 to 3 days when water temperatures are low.

Test Animals

In tests where the biologist must determine 

the ability of fish to withstand exposure to a 

toxicant in order to ensure selection of a safe 

concentration for treatment (e.g., removal of 

scalefish from a catfish pond), use of fish from 

the pond to be treated is mandatory. In other 

situations as well, test animals from the waters 

to be treated should be used if they can be 

collected in good condition and in adequate 

numbers. If this is not possible, target and 

nontarget fishes of appropriate sizes must be 

brought in from other sources and allowed to 

acclimate for 24 h in live-cages before the test. 

Sizes and numbers needed depend on the 

loading levels that can be used under conditions 

existing at the time of the test.

Loading Rates

The following information provides broad 

guidelines regarding loading rates. At temperatures of 25 to 30 C, up to a 3-g/l load (852 g per 

bag) of fish can be used for 96-h tests. For each 

5-degree reduction in temperature below 25 C, it 

may be possible to increase the loading by 1 g/1 

(284 g per bag). For 24-h tests, these amounts 

can be increased, depending upon the circumstances. If excessive loading rates are used, 

results of the test are likely to be too biased to be

usable. Temperature, dissolved oxygen, and pH 

should be measured each morning and afternoon throughout the test period.

Duration of Toxicity Tests

Test results are apparent much sooner at high 

than at low temperatures, and the concentration 

of antimycin required for a complete kill in 

warm weather can safely be selected on the 

basis of a 24-h test. In cold weather such tests 

should be conducted for at least 48 h to avoid the 

use of unnecessarily high concentrations of 

toxicant.

Selection of concentrations to effect partial 

kills can be based on 24-h tests only if temperature and pH are high enough to ensure complete 

detoxification of the antimycin within that 

time. When temperatures are low and the pH is 

neutral or acid, tests for partial kills should be 

carried out for 96 h or until the full effects of the 

various concentrations are discernible.

Selecting Treatment 

Concentrations

The results of on-site toxicity tests can be used 

in selecting treatment concentrations if two 

conditions are met: (1) mortality patterns must 

be consistent, i.e., mortalities must increase as 

toxicant concentrations increase, and (2) mortality of the control fish must not exceed 10%. 

Inconsistent patterns of mortality often are a 

result of using poor quality test animals. If test 

results are good, the lowest concentration at 

which all target fish are killed should be used 

when the entire volume of water is treated to 

effect a selective kill. The concentration of 

toxicant used in each bag can be calculated 

precisely because the volume of water is known. 

Success in applying test results to actual 

treatments is directly proportional to the accuracy with which the volume of water to be 

treated is determined.

When a complete kill is intended and large 

specimens of the target species are used in the 

toxicity test, the lowest completely effective 

concentration tested should be increased by at 

least 1.0 j-cg/1 to ensure effectiveness of the 

treatment. If large specimens are not used, a 

safety factor of at least 1.5 to 3.0 //g/1 should be 

added to the lowest completely effective 

bioassay concentration, depending on the 

susceptibility of the target species.
LITERATURE CITED

Berger, B. L., P. A. Gilderhus, and R. E. Lennon. 1967. 

Attempted reclamation of Whitewater Lake, Valentine 

National Wildlife Refuge, Nebraska. Fish Control 

Laboratory, U.S. Bur. Sport Fish. Wildl., La Crosse, 

Wisconsin. 12 pp. (Mimeo)

Gumming, K. B., R. M. Burress, and P. A. Gilderhus. 1975. 

Controlling grass carp(Ctenopharyngodon idellaVal.) by 

chemical and physical means. Prog. Fish-Cult. 

37(2):81-84.

Gumming, K. B., and P. A. Gilderhus. (In press). Posttreatment effects on water quality and nontarget 

organisms following piscicidal use of antimycin. Trans. 

Am. Fish. Soc.

Davis. J. T., and W. S. Hardcastle. 1959. Biological assay of 

herbicides for fish toxicity. Weeds 7(4): 397-404.

Falk, M. R. 1972. Simple apparatus for conducting acute 

toxicity bioassays under field conditions. Water Res. 

7:821-822.

Fremling, C. R., and J. J. Evans. 1963. A method for

determining the dissolved oxygen concentration near the

mud-water interface. Limnol. Oceanogr. 8(3): 363-364. 

Howell, J. H., and W. M. Marquette. 1962. Use of mobile

bioassay equipment in the chemical control of sea

lamprey. U.S. Fish Wildl. Serv., Spec. Sci. Rep. Fish. 418.

9pp. 

Lennon, R. E., and P. S. Parker. 1959. The reclamation of

Indian and Abrams Creeks in Great Smoky Mountains

National Park. U.S. Fish Wildl. Serv., Spec. Sci. Rep. Fish.

306. 22 pp. 

Pfeiffer, P. W. 1968. Small lake studies. Ky. Dep. Fish Wildl.,

Fed. Aid in Fish Restoration, Job Prog. Rep. F-30-R-1,

Job No. 1-F: 44-47. 

Vaughan, T. L., M. Quick, K. Denson, and D. Parrish. 1974.

Suwannee Lake (Rep. 2), pp. 1-22 in 1973 Fish Manage.

Rep. NE Region. Fla. Game Fish Comm. Lake City, Fla.

Feb., 1974.
Table 1. Temperature, pH, and dissolved oxygen ranges in ponds where on-site 

bioassay experiments were conducted.

Date

4/69

5/69

5/69

5/69

5/69

6/69

7/69

8/69

11/69

11/69

11/69

11/69

11/69

12/69

1/70

3/70

4/70

6/70

6/70

Test 

period

(h)

99

70

70

107

83

72

72

72

96

96

120

120

72

72

81

96

96

96

96

Loading 

level

(g/D

0.75

0.93

0.68

0.45

0.55

0.31

a

1.41

0.46

1.23

0.62

1.24

0.99

0.89

2.76

0.85

0.11

0.49

0.55

Surface 

temperature 

range

(°C)

17.8-23.4

21.7-30.6

18.9-27.2

22.2-30.0

23.4-30.6

29.5-35.0

26.1-31.7

26.7-31.1

13.7-16.2

13.7-16.2

11.2-14.5

11.2-14.5

8.9-12.2

6.8- 8.9

8.9-15.6

12.2-15.6

17.8-21.1

23.4-26.1

25.0-31.7

pH range Dissolved oxygen range (mg/l)

Plastic bags

Pond

6.7- 8.2

6.7- 8.2

6.8- 8.4

6.7- 8.9

6.6- 9.3

7.3- 8.9

7.6-10.3

6.6- 8.3

5.9- 6.4

5.9- 6.4

5.8- 6.3

5.8- 6.3

7.0- 7.3

7.3- 7.6

7.3- 8.8

5.3- 6.4

6.8- 7.5

7.5- 8.9

7.0- 9.3

Fish

6.8- 8.0

6.3- 6.8

6.9- 8.5

6.6- 7.6

7.2- 9.4

7.2- 9.0

7.9-10.4

6.6- 8.5

5.1- 6.6

5.5- 6.1

6.3- 6.8

6.0- 6.2

6.6- 7.0

7.3- 7.4

6.8- 8.6

5.9- 6.1

6.8- 7.5

7.4- 8.7

7.0- 8.6

No fish

7.2- 8.1

6.7- 7.3

7.2- 8.7

6.6- 8.9

7.8- 9.4

7.8- 9.1

7.9-10.3

6.6- 8.3

6.1- 6.5

6.1- 6.5

6.3- 6.8

6.3- 6.8

7.2- 7.4

7.3- 7.4

6.0- 6.1

6.8- 7.2

7.7- 8.8

7.0- 8.3

Pond

6.6-10.0

5.4- 9.6

6.3-10.5

6.1-13.0

5.4-13.5

6.8-10.7

3.0-14.0

6.1- 9.0

8.8- 9.8

8.8- 9.8

7.4-10.0

7.4-10.0

10.2-13.3

10.0-14.0

11.4-14.5

5.0- 8.2

6.6-10.4

8.6-12.2

6.2-11.2

Plastic bags

Fish

6,0- 9.5

0.5- 6.5

5.7- 9.2

3.3-10.4

8.2-14.1

7.5-10.3

6.2-24.0

5.2- 8.7

9.2-10.0

5.0-10.6

7.8- 9.0

4.7- 6.3

10.4-11.2

12.6-13.2

8.6-13.8

8.6-10.4

8.0-12.4

8.5- 9.8

No fish

7.5- 9.5

4.9- 8.2

5.9-10.3

5.8-14.3

7.2-15.4

7.7-10.8

8.6-19.9

6.5- 9.0

9.6-10.0

9.6-10.0

8.6-10.4

8.6-10.4

10.8-12.8

13.5-13.6

8.4-10.5

9.4-12.2

8.1-11.2

a Turtles damaged bag, allowing fish to escape.
Figure 1. Sample apparatus for suspending large plastic bags used to conduct on-site toxicity tests.


(Reports 53 through 55 are in one cover.)

53. Toxicity of Mixtures of Quinaldine Sulf ate and MS-222 to Fish, by Verdel K. Dawson and Leif L. 

Marking. 1973. 11 pp.

54. The Efficacy of Quinaldine Sulfate:MS-222 Mixtures for the Anesthetization of Freshwater 

Fish, by Philip A. Gilderhus, Bernard L. Berger, Joe B. Sills, and Paul D. Harman. 1973. 9 pp.

55. Residues of Quinaldine and MS-222 in Fish Following Anesthesia with Mixtures of Quinaldine 

Sulfate:MS-222, by Joe B. Sills, John L. Alien, Paul D. Harman, and Charles W. Luhning. 1973. 

12pp.

(Reports 56 through 59 are in one cover.)

56. Toxicity of the Lampricide 3-trifluoromethyl-4-nitrophenol (TFM) to 10 Species of Algae, by A. 

A. Maki, L. D. Geissel, and H. E. Johnson. 1975. 17 pp.

57. Actue Toxicities of 3-trifluoromethyl-4-nitrophenol (TFM) and 2',5-dichloro-4'-nitrosalicylanilide (Bayer 73) to Larvae of the Midge Chironomus tentans, by J. A. Kawatski, M. M. 

Ledvina, and C. R. Hansen, 1975. 7 pp.

58. Acute Toxicity of the Lampricide 3-trifluoromethyl-4-nitrophenol (TFM) to Nymphs of 

Mayflies (Hexagenia sp.), by C. R. Fremling. 1975. 8 pp.

59. Toxicity and Residue Dynamics of the Lampricide 3-trifluoromethyl-4-nitrophenol (TFM) in 

Aquatic Invertebrates, by H. O. Sanders and D. F. Walsh. 1975. 9 pp.

(Reports 60 through 62 are in one cover.)

60. Toxicity of the Lampricide 3-Trifluoromethyl-4-nitrophenol (TFM) to Nontarget Fish in Static 

Tests, by L. L. Marking and L. E. Olson. 1975. 27 pp.

61. Toxicity of the Lampricide 3-Trifluoromethyl-4-nitrophenol (TFM) to Nontarget Fish in FlowThrough Tests, by L. L. Marking, T. D. Bills, and J. H. Chandler. 1975. 9 pp.

62. Toxicity of the Lampricide 3-Trifluoromethyl-4-nitrophenol (TFM) to Selected Aquatic 

Invertebrates and Frog Larvae, by J. H. Chandler and L. L. Marking. 1975. 7 pp.

(Reports 63 through 66 are in one cover.)

63. Laboratory Efficacy of 3-Trifluoromethyl-4-nitrophenol (TFM) as a Lampricide, by V. K. 

Dawson, K. B. Gumming, and P. A. Gilderhus. 1975. 7 pp.

64. Effects of 3-Trifluoromethyl-4-nitrophenol (TFM) on Developmental Stages of the Sea 

Lamprey, by G. W. Piavis and J. H. Howell. 1975. 4 pp.

65. Accumulation and Loss of Residues of 3-Trifluoromethyl-4-nitrophenol (TFM) in Fish Muscle 

Tissue: Laboratory Studies, by J. B. Sills and J. L. Alien. 1975. 5 pp.

66. Residues of 3-Trifluoromethyl-4-nitrophenol (TFM) in a Stream Ecosystem after Treatment for 

Control of Sea Lampreys, by P. A. Gilderhus, J. B. Sills, and J. L. Alien. 1975. 5 pp.

67. Method for Assessment of Toxicity or Efficacy of Mixtures of Chemicals, by L. L. Marking and 

V. K. Dawson. 1975. 7 pp.

Fish Control Laboratories

Fish and Wildlife Service

U.S. Department of the Interior

P.O. Box 862 

La Crosse, Wisconsin 54601

GPO 832- 110
As the Nation's principal conservation agency, the Department of the 

Interior has responsibility for most of our nationally owned public 

lands and natural resources. This includes fostering the wisest use of 

our land and water resources, protecting our fish and wildlife, preserving the environmental and cultural values of our national parks and 

historical places, and providing for the enjoyment of life through outdoor recreation. The Department assesses our energy and mineral 

resources and works to assure that their development is in the best 

interests of all our people. The Department also has a major responsibility for American Indian reservation communities and for people who 

live in island territories under U.S. administration.

UNITED STATES

DEPARTMENT OF THE INTERIOR

FISH AND WILDLIFE SERVICE

WASHINGTON. D. C. 2O24O

POSTAGE AND FEES PAID 

U.S. DEPARTMENT OF THE INTERIOR

INT 423