Biology of Freshwater Fish in a Review of the Literature of 1967 On

Home , Sevan trout

Biology of Freshwater Fish

A Review of the Literature of 1967

Wastewater and Water Pollution Control

Max Katz and Albert K. Sparks College of Fisheries University of Washington, Seattle

with

Gary L. Pederson, C. E. Woelke, James Woodey College of Fisheries University of Washington, Seattle

To be published in

Journal Water Pollution Control Federation Volume 40, No. 7, July 1968

Prepared for

The Research Committee of the Water Pollution Control Federation You will notice that this prepublication manuscript is entitled "Biology of Freshwater Fish." Previous to this year, the section of the review prepared at the College of Fisheries,

University of Washington, had included also sections on algae, fungi, radioactive wastes, and the biological aspects of marine pollution. The explosion in the literature on the biological aspects of water pollution has made it advisable to divide the review of the biological aspects of water into several smaller sections, some of which were assigned to other reviewers at different institutions and agencies.

The Editors of the Journal of the Water Pollution Control

Federation are collecting and publishing all of the sections of the Literature Review in one issue. The July issue of the

Journal will be the Literature Review issue. This can be pur- chased from the Water Pollution Control Federation, 3900 Wis- consin Avenue, Washington, D.C. 20016. In my opinion, this would be worth getting.

Max Katz Errata, 1967

P. 5, line 12: delete "fish". P. 7, lines 11 and 12: should read "decomposed" not "decomposited". P. 8, line 1: should read "LAS, mortalities". P. 9, par. 2, line 4: should read "least sensitive, goldfish". P. 10, line 3: to read "even after the fish has been washed with tap- water" etc. P. 11, line 2: delete "(same ref. as above)". P. 11, line 7: delete "(same ref. as above)". P. 14, par. 1, line 7: to read "roach, Leuciscus rutilus, were". P. 16, par. 1, line 11: change 15 ppm to "15 mg/kg". P. 19, par. 2, line 6: delete "breathing" and substitute "respiring". P. 26, line 9: should read "of" not lpf". P. 29, line 13: delete "weed" and insert "herbivorous". P. 30, par. 1, line 10: delete "effects" and insert "components". P. 30, par. 1, line 11: change "concentration" to "concentrations". P. 32, line 2: insert "the" between "for" and "mortality". P. 33, par. 2, line 6: delete "to" and insert "which will". P. 33, par. 2, line 7: delete "to" and insert "which will". P. 35, line 12: last word should read "stretch". P. 40, par. 3, line 5: insert "The effects of" between "carp" and "Introduction". P. 40, par. 3, line 7: delete "tested" and substitute "observed". P. 42, par. 1, line 4: delete "increasing" and substitute "increased". P. 49, ref. 39, line 2: should read "Turtox". P. 50, ref. 47, line 2: should read "aver" not "Over". P. 55, ref. 77, line 2: should read "Acetylcholinesterase". P, 55, ref. 78, line 2: insert "Patent" between "U.S." and "3,271,246". P. 62, ref. 128, line 2: after "Rill." insert "31st N. American Wild- life & Nat. Res. Conf., Pittsburgh, Pa. 11 p. P. 70, ref. 178, line 3: "Zambia" not "Zanbia". General Aspects of Water Pollution Biology

Perhaps the most important recent single contribution to the literature on the biological aspects of water pollution is the publication by Keup, Ingram and Mackenthun (96) of a collection of important papers on water pollution. The papers included, date as far back as Kolkwitz and Marsson's (1908) classical paper, yet include some papers published in the 1960/s. It is probable that this compilation will become a standard reference for all persons interested in the biological aspects of water pollution, and it is certain that there will be a long and continued demand for this book.

Mills et al. (124) have documented the biological changes in the Illinois River during the past seventy years that have resulted from man's activities. The changes in the biota of the river since the late 1890's have been variable; some organisms have responded favorably but, in general, there has been a drastic and undesirable reduction in the numbers and kinds of desirable plants and animals that live on or in the river.

Zhilenko (202) reported that the 1962 fish catch in the Suifan

River was 247 of its 1942 catch. The decline in fish abundance was due to domestic pollution and to overfishing.

The effects of pollution, especially municipal and industrial wastes, upon aquatic life in streams are detailed by Ingram and

Mackenthun (81). Pollution may stimulate associations of organisms that are nuisances to man. The biological responses of 2 organisms to various wastes are illustrated. Among the types of pollution and their effects upon biota that are illustrated in this review are the effects of siltation in the Bear River of

Utah; sewage and textile wastes in the Chattooga River, Georgia; pulp and paper wastes in the Menominee River in Michigan and Wiscon- sin; pulping and chemical wastes in the Pearl River in Mississippi; and acid mine discharges upon the Monongahela River in Pennsylvania.

A list of parameters that can be considered in setting water quality standards are presented.

In addition, Ingram et al. (82) has compiled a valuable publication that is a combination textbook, bibliography, and glossary of biological organisms. Ingram's publication is of value to biologists, chemists, and engineers wishing to develop a background of knowledge in the biological response of organisms to pollution.

The second edition of the "Guide to the Biology of Domestic and

Industrial Water Supplies" has been published in Germany (9). It includes chapters on biological sampling procedures, methods of biological examination, biological indicators of water quality, in addition to material on water supply problems.

Malacea (116) stated that two subject areas are important in the study of river pollution, (1) the knowledge of the toxicities of various substances and their mixtures, and (2) the knowledge of the influence of toxic substances on the processes of self- purification. 3

A fish kill involving from 300 to 500 tons of fish was observed in the Elbe during the winter of 1962/1963. The ice cover of the river prevented reoxygenation; low-water levels and high concentrations of ammonia and phenols contributed to the problem

(102). Water quality conditions leading to a fish kill in the

River Kali were discussed by George at al. (60). Sigler and co- workers (161) described the effects of uranium mill wastes on insects, fish, and algae. Nehrkorn (133) stressed that the basis of all biological investigations is the community of flora, fauna, and micro-organisms. These give an overall picture of the nature and quality of the water which has passed over a known place for a long period of time. Rothschein (153) suggests a new method for calculating an index of biological water quality which has been used successfully in the study of various rivers in Czechoslovakia.

Fish kills caused by phytoplankton blooms are discussed by Swingle

(177). Water Quality and DO Requirements of Fish

The necessity imposed upon the various states to submit water quality criteria for approval by the Federal Water Pollution Con- trol Administration has stimulated a great deal of interest in the water quality requirements of fish. The discussions of the water quality requirements of fish by Stroud (174), Wurtz (198),

Tarzwell (179), and Surber (176) are typical of the literature resulting from this stimulus. Stroud emphasized that water quality must be sufficiently high to provide for the protection and maintenance of vigorous populations of aquatic life, especially fishes. He also stressed that the exact numerical criteria for many 4 environmental factors are not known at the present time, and that adequate safety factors must be allowed to compensate for variations due to local conditions. Wurtz (198) stressed that proper waste disposal can be compatible with the use of water for aquatic life.

Surber (176) emphasized that turbidity, water temperature, and toxicants be considered in the formulation of water standards for fish.

Doudoroff and Shumway (43) feel that the complete protection of fish will not allow for the reduction of dissolved oxygen below saturation levels. Adequate growth, embryonic development and the activity of fish can be limited by a reduction of DO only slightly below the saturation limits. DO criteria should be based on considerations other than that of survival. Stewart et al. (170) studied the effects of constant dissolved oxygen concentrations and of wide diurnal fluctuations of oxygen concentration on the appetite, growth, and food conversion of largemouth bass.

At constant dissolved oxygen concentrations, food conversion efficiencies were markedly reduced only below 4 mg/1 DO. The growth of bass subjected alternately to low and high oxygen concentrations for each 24 hr day was markedly impaired.

Strelttsova (172) observed the adaptation of trout and carp to various concentrations of dissolved oxygen. At low temperatures,

1° to 2°C, trout adapted after 22 days. At higher temperatures,

16 to 17°C, both species adapted more rapidly. Galkina (58) studied the effects of mechanical agitation and oxygen deficiencies 5 on the developing eggs and larvae of lamprey. The tolerance of larvae to an oxygen deficit was highest 8 to 12 days after hatching. The tolerance to an oxygen deficity diminished with age and the transition to active feeding. Hoff et al. (75) in experiments with marine and anadromous fish showed that the lethal oxygen level was partially dependent upon temperature and stress.

Reid (151) observed a mass mortality in a Florida lake which was due to oxygen depletion to a level less than 20 percent of saturation. This low level was maintained for more than a week.

Fotiev (54) claimed that deoxygenated groundwater entering the

Rybinsk Reservoir was responsible for fish kills. Haley et al.

(65) reported a fish kill of shad that was due to the stress of a bacterial infection superimposed on a low oxygen condition.

Heath and Pritchard (69) subjected bluegill sunfish and cut- throat trout to a rapidly declining oxygen tension until they began to lose equilibrium. During recovery from stress, measurements of changes in their blood chemistry were made. The return to normal levels was delayed in the trout as compared to the bluegill. Gottwald and Kaniewski (62) observed the effects of reduced dissolved oxygen on the functioning of the blood system of trout.

Hematomas were observed to develop where the blood vessels were thin or poorly developed. These hematomas prevented the heart from circulating the blood adequately. Ryzhkav (155) studied the tolerance of three forms of the Sevan trout to temperature, reduced DO, and direct sunlight. The requirements of each of the 6 several developmental stages was different for each form.

Pauley and Nakatani (139) investigated the histopathclogy of chinook salmon fingerlings which had gas-bubble disease. The most striking changes occurred in the roof of the mouth of all fish while many of the other organs with the exception of the hearts and stomachs exhibited some changes. Schofield (158) observed that there is a relation between survival of brook trout and water quality. Heavy mortality of planted trout which had come from both hard-water and soft-water hatcheries coincided with periods of high acidity and high concentrations of heavy metals in the waters of Lake Honnedaga, N.Y. during the summer, but trout from another hard-water hatchery where the water supply contained traces of zinc, survived transfer to the lake. Ivasik

(83) found that alkaline water (pH 10 to 11) harmed fish while pH 8.5 to 9.0 prevents infectious dropsy in carp. Silvey (162) found that the high carbon dioxide content and related low pH of water caused a high mortality of salmon and steelhead eggs and fry at the Trinity River Fish Hatchery near Lewiston, Calif.

To reduce the carbon dioxide content, limestone chips were placed in the intake works of the hatchery. The maintenance of the pH between 7.5 and 8.0 has served to reduce losses.

Low DO was believed by Tait (178) to be responsible for two large fish kills observed in the Kafue River, Zambia. It was believed that local conditions of low DO forced fish into shallow waters where the oxygen was quickly depleted. 7

Domestic Wastes

Rogers (152) made an extended investigation of the inver- tebrate and fish populations and water quality in two streams which received much of the domestic wastes of Auburn, Alabama.

The biological changes resulting from the introduction of the domestic wastes were recorded. Water quality determinations clearly showed the polluted conditions of the streams and the recovery following abatement procedures. Ray and David (149) surveyed 35 km of the Ganges River to study the effect of sewage and industrial wastes upon fish populations. Complete deoxygen- ation does not occur because of the rapid currents of the river.

Fish populations are depressed below the outfalls but the decomposited wastes fertilized the river and eventually stimulated the growth of aquatic organisms. Allen and O'Brien (4) experimented with juvenile chinook salmon in saline waters mixed with effluent from oxidation ponds to determine if sewage effluent could be used to increase natural foods and increase growth. The tolerance of salmon to saline waters varied at different ages.

Detergents

Thatcher and Santner (182) determined the toxicity of a typical linear alkyl sulfonate detergent to five species of freshwater fish in a flowing water system. Lake emerald shiners were the most sensitive while the black bullheads were the most tolerant. In tests with ABS most of the mortality occurred during 8 the first two days of the test but with LAS mortalities also occurred on the 3rd and 4th days. Chwala (29) observed that the toxic effect upon fish of synthetic detergents is non-specific and results from the depression of surface tension. When the surface tension of the water is reduced to 50 dynes/cm, the epithelial cells of the fish gills become irreversibly swollen and distorted. Biodegradable detergents are less likely than branched-chain sulfonates to reach harmful concentrations in natural waters. Lang (103) made histological and cytological examinations of the injuries caused by anionic surface active agents to fish, and concluded that the lytic and hemolytic properties of these substances will kill fish. The toxicity of these syndets as well as the degree of the histological and cytological changes and the symptoms of poisoning can be influenced by the salt composition of the medium. Most histological changes are observed in the gills and epidermis and only slight injuries can be observed in liver and kidney tissues. Pfeiffer (142) detected changes in the olfactory and gustatory organs of catfish which had been exposed to a concentration of 0.5 to 10 mg/1 of detergents at a temperature of 20°C. Scheier and Cairns (157) observed that gill damage in sunfish resulting from a 24 hr exposure to 18 ppm ABS was not reversible even after the fish were kept in clean water for eight weeks. Mann (117) studied the physical and chemical properties of water as they relate to fish toxicity. He intensively studied the toxicity of dodecylbenzene- sulfonate to rainbow trout with special reference to changes in 9 temperature, calcium concentration, DO and pH, all of which may have an effect on the toxicity of the detergent to fish.

Industrial Wastes

Veszprgmi (188) investigated the damage to fish populations caused by pollution from dairies, distilleries, piggeries, and chemical industries. Fish were found to survive for 18 to 20 min in the wastes from a fertilizer plant which contained 4 mg/1

NH4 and 0.07 mg/1 NO2 (51). Westoo (194) determined the concentration of Hg in several species of North Atlantic marine fish, and in freshwater fish. With some exceptions, marine fish contained less mercury than the FAO standard. It was observed that the larger specimens, contained more Hg than smaller specimens from the same location. The highest concentrations of Hg were found in freshwater fish which were taken near the Swedish Coast, and was probably due to industrial pollution because the same species ta n from non-polluted or slightly-polluted interior Lakes contained much less Hg.

Costa (33) found that cyanide is highly toxic to all species of fish but that fish could detect and avoid NaCN at concentrations -6 as low as 10 N. The most sensitive fish were trout fry and the least sensitive goldfish. The intensity of the response is affected by temperature, pH, and DO. Additional experiments were conducted with the crustacean Gammarus and the tadpoles of Rana. Environ- mental conditions also affected the reactions of these organisms. 10

Kariya et al, (88) conducted studies on the detection and evaluation

of cyanides in the tissues of fish killed by these substances0 Ii

was possible to detect cyanides even after washing with tap water

for 48 hr after death. Leduc (105) studied the physiological and

biochemical responses of fish to chronic exposure to cyanides.

David and Ray (38) concluded that a danger to fish life was present

at all seasons in the river Daha which receives sugar and distillery

wastewaters. Despite the pollution, fish moved into the Daha and

thrived for short periods in the less polluted areas until a flushing

of materials from the sugar mills killed them. Brovko and Chernyi

(19) and Yarovenko et al. (199) described their experiences in

rearing carp in the waste lagoons of two separate Russian sugar

refineries. The wastes were treated with lime, NH4NO3 and super-

phosphate, and the growth of the fish seemed to be satisfactory.

l Luk yanenko, Flerov, Volodin and their associates (110), (111),

(53) and (189) conducted a long series of observations upon the

effects of phenols on fish. The resistance of Carassius to

several toxic phenol concentrations did not change with the gradual

increase of water hardness. Solutions of pH 3 or 12 were lethal

to the fish, but pH 4 or 11 did not affect them. The resistance

of the fish to a concentration of 50 mg/1 phenol was not affected

by pH changes within the range of 4 to 11. The resistance of fish

to the minimum lethal concentration of phenol did not change in the

temperature range of 5 to 25°C. Young trout, one-half-year-old, were more tolerant to phenol than were two-year-old rainbows and brown trout. 11

The eggs of Abramis were incubated in various concentrations of phenol by Volodin et al. (189) (same ref. as above) and the pathology of the eggs in each concentration was described. The maximum tolerable concentration of phenol was 25 mg/l. The changes in the activity of cholinesterase in the muscles and of ammonia in the brains of phenol-poisoned Carassius were measured by Luklyanenko and Petukhova (111) (same ref. as above). The stimulating and paralyzing effects characteristic of phenol were followed by the inactivation of cholinesterase, and the consequent accumulation of acetylcholine in cholinergic synapses.

Pulping Wastes

Biological observations by Thomas and Legault (183) indicated that the waste treatment processes of a Michigan paper products plant were effective. An abundant fish fauna vas found below the effluent of the plant. The minnow, Notropis, was found to be intolerant of low concentrations of untreated effluent; its presence, therefore, in the area was an indication of effective treatment procedures. Hasselrot (67) placed young salmon, pike, and perch, in boxes below the effluent of two kraft mills. At both mills, there was evidence that the effects of the wastewaters extended for a considerable distance from the point of discharge and the effects were more noticeable in winter owing to the slower rate of decomposition of the waste in the colder water. The author concludes that live-box experiments are useful for detecting pollution by pulp mills. 12

Colby and Smith (30) determined the water quality characteristics of water which lay over fiber deposits below paper mills.

The gradient of DO from the sludge water interface to the water surface varied from 0.1 mg/1 to near saturation. The dissolved sulfides varied from 0.8 mg/1 to zero. The most rapid gradient was in the 20 mm of water just above the fibers. Walleye eggs placed in test trays in fiber mats had a lower survival than those placed 12 in. above the mats, and those placed on mineral sub- strates. Laboratory experiments showed that sulfide levels of

0.3 mg/1 were acutely lethal to Gammarus and to walleye eggs and fry.

Metals

McDonald and Gaufin (112) found that the use of copper sulfatg as an algicide does not harm fish, provided the chemical is applied uniformly in concentrations of 1 mg/1 to areas of open water.

Uneven application of copper sulfate, especially in shallow areas of water, may result in damage.

Beyerle and Williams (13) attempted control of bluegill reproduction by the application of copper sulfate crystals to spawning nests. Experimental variability was so great that no definite conclusions could be made in regard to the efficacy of the treatments.

Kariya et al. (89) were able to detect and measure the quantity of copper in fish which had been killed by wastes containing copper. The effect of various concentrations of cobalt on development of eggs and the growth of fingerlings, yearling 13 and two-year-old rainbow trout was investigated by Shablina (159).

Telitchenko (181) studied the long-term effect of elevated concentrations of uranium on the fertility and survival of guppies.

Kariya et al. (90) used the diphenyl-carbazide method to detect chromium in the bodies of fish killed by chromium solutions. Berg- lund and Wretlind (10) studied the toxicologic levels of mercury in Swedish fish. Persons eating 200 g of fish from Hg-polluted water had a blood concentration of Hg 20 to 30 times greater than normal. The highest concentration of fig in fish observed in Sweden was 8.0 mg/kg. Fish containing 1 mg Hg/kg or more should be considered unsuitable for human consumption.

Bluegills were exposed to sublethal concentrations of cadmium sulfate for periods up to 90 days by Mount and Stephan (127). In living fish, accumulation of cadmium in gill tissue never exceeded

130 ug/g based on dry weight, but fish that died of acute cadmium poisoning contained a minimum of 150 ug/g in their gill tissue. The data suggests that cadmium poisoning can be detected by measuring cadmium in the gill tissue.

Grande (63) carried out laboratory experiments on the effects of copper and zinc on the eggs, fry, and fingerlings of Atlantic salmon, brown trout, and rainbow trout. In the experiments with fry in the yolk-sac stage there was evidence that brown trout were slightly more tolerant to both metals than were rainbow trout and salmon. It was found that small amounts of copper and zinc reduced the percentage of eggs hatching. Saunders and 14

Sprague (156) reported that copper and zinc pollution from a mine on a tributary of the Northwest Miramichi River caused many adult Atlantic salmon, which were on their normal upstream spawning migration, to return prematurely downstream through a counting fence during summer and early autumn. These observations gave an opportunity to document avoidance reactions of salmon to pollution, which has seldom been done in the fishes 1 natural environment.

Oxygen uptake by zebrafish (Brachydanio rerio) in relation to zinc sulphate resistance was measured by Skidmore (163). A math- ematical relationship between the rate of oxygen uptake and survival time is postulated. Lange (104) investigated the effect of zinc on the phagocyte reaction of leukocytes in carp and goldfish blood. Seasonal variations in copper, manganese, iron, and zinc, in the organs and tissues of roach Leuciscus rutilus, was determined by Ilzina (80). In all organs the content of trace elements was considerably higher towards the end of the vegetation period (August), when adequate varied food was available, than in spring (May).

Pesticides

Cope (32) discussed the contamination of freshwater ecosystems by pesticides, and the subsequent fate of the pesticides.

The acute effects of pesticides on the fauna and flora are examined. Sub-acute effects, such as those on behaviour, growth, mortality, reproduction, pathology, resistance, and genetics are reviewed. It is emphasized that the variety of environmental 15 features of lakes and streams, the multiplicity of chemicals and formulations, result in interactions that can be highly dynamic and difficult to predict.

Organochlorine residues in the Green Bay area of Lake

Michigan were investigated by Hickey at al. (72). Analyses were made of: (a) shallow and deep water mud samples, (b) the amphipod, Pontoporeia affinis, which is an important food source for fishes, (c) alewives, whitefish, and chub, and (1) oldsquaw ducks, ring-billed gulls, and herring gulls. The biological concentration of pesticides, similar to those previously reported for smaller lakes, was reported. In Pontoporeia the concentration factor is about 50 times of the residue level of the mud, and in fish this factor is increased about 10 times more. The persistence of toxaphene in lakes was the subject of an investigation by

Johnson et al. (86). Lakes treated with 0.1 mg/1 of toxaphene

3 to 9 yr prior to sampling had the following distribution of toxaphene: water 1 to 4 ppb, sediments 0.2 to 1 mg/1, and aquatic plants 0.05 to 0.4 mg/kg. Thirty-five farm pond ecosystems in southwestern Virginia were examined for heptachlor residues (192).

Every pond watershed was found to contain heptachlor residues, although in amounts 4.300 ppb. Pond water was found to contain residues only in the spring, while soil residues decreased during the summer months. Residues of heptach/or were detected up to

25 months after application. 16

Edwards (46) studied the effect of TDE on the fauna of a pond. On the basis of laboratory tests, TDE was considered a more useful insecticide than DDT because of its lower toxicity to fish.

A comparison of the organisms in a treated pond (1.24 kg/ha of TDE) and a neighboring control pond showed that mud-dwelling chironomid larvae were greatly reduced in species and numbers, and that culicids and ephemeropterans were also killed. There was no discernible effect on other benthic or planktonic organisms. Carp were kept in cages for 11 months following the insecticide treatment; 35 percent died but the survivors grew well. By the 11th month, the tissue of the fish contained about 15 ppm TDE.

Sparr at al. (166) determined the concentration of insecticides in waterways, fish and the mud from various fields which had been treated with endrin and aldrin. Runoff water from a cotton field, treated 3 times with endrin at .372 kg/ha showed only low concentrations of endrin and only 0.05 mg/1 after the last spraying. Traces of endrin were found in fish but none in mud.

Residues in water from corn fields (6.2 kg/ ha/yr) or waterways draining rice fields (.31 kg/ha) did not exceed a few parts per thousand.

Residues in fish from the waterways draining rice fields rarely exceeded 10 ppb/aldrin plus dieldrin. 14 Water translocation of C-labe1ed diazinon off a model cranberry bog was studied by Miller et al. (123). Transportation of 14 35 diazinon- C and parathion- S was shown 24 hr after application.

The majority of the chemicals disappeared from the water after 17

144 hr. During this time, no labeled metabolites of diazinon were detected; however, 3 labeled parathion degredation products were encountered. Accumulation of both pesticides occurred in the fish and mussels. The fish appeared able to metabolize these substances at a faster rate than the mussels.

There have been several studies reporting the effects of forest sprayings with insecticides. In the first of two papers,

Kerswill (94) outlined (a) the history of the spray program developed to counteract an outbreak of spruce budworm in New Brunswick;

(b) administrative arrangements necessary for a research and development program concerning the affected fisheries, mainly

Atlantic salmon; (c) highlights of field and laboratory investigations regarding the effects of insecticide sprayings on the fauna of the streams. In his second paper, Kerswill (95) spec- ifically discussed the fish losses resulting from the sprayings.

DDT-in-oil in a single application, 620g/ha, caused heavy losses of underyearling salmon and parr within 3 wk. DDT in oil at

310g/ha had no apparent short-term effects on salmon parr, but killed many underyearlings.

Elson (47) who also studied the effects upon salmon of DDT spraying on New Brunswick's forests, got results similar to those described by Kerswill. Elson also found evidence of a delayed mortality that occurred 4 to 6 months after spraying. It was associated with the onset of winter cold. In a paper written together, Elson and Kersuill (48) discussed the impact on salmon 18 of spraying insecticides over forests. They showed that spraying had adverse effects on fish and fish food organisms, and there was evidence of delayed mortalities and that sub-lethal concentrations of DDT can have harmful effects on fish. Spraying programs have been modified in an attempt to reduce the amount of insecticide falling on streams, and increasing use is being made of Phosphamidon.

The use of Phosphamidon at 620g/ha near streams has not caused any marked reduction in indigenous fish populations. Cole et al. (31) made a comparison of DDT levels in fish, streams, stream sediments, and soil before and after a DDT aerial spray application in northern

Pennsylvania.

Nicholson (135) presented a lengthy discussion of water pollution by persistent insecticides and their effects on aquatic fauna, particularly fish. He proposed utilizing collected data regarding the effects of these substances upon aquatic forms to obtain meaningful water quality standards. The consequences of insecticide use on non-target organisms was the subject of a review

(134). Fish are among the non-target organisms discussed. Since fish can concentrate chlorinated hydrocarbons up to 10,000 times, the water quality criteria for these substances should be based upon this biological magnification and not upon the TL1m (50).

Mount (125) suggested that the best and most direct measure of adverse effect from exposure to pesticides is the production rate

(including growth and reproduction) of the animals one wishes to harvest. However, establishment of tolerable concentrations of 19 pesticides for fish requires the consideration of (a) food-chain accumulation; (b) tissue residues rendering the fish unfit for consumption; (c) potential hazard to the fish from readsorption of fat-stored pesticides; and (3) off-tastes or tainting from certain types of pesticides.

A summary report (128) of the studies in the lower Mississippi

River described sources, mechanism of transport, and modes of action of the implicated pesticides. Novak and Ramachandra (137) described the program in the lower Mississippi of monitoring endrin in fish and shellfish that might be consumed by humans.

A list of insecticides and herbicides used in Japan and their toxicity to rainbow trout, carp, and some other Japanese fish was published by Kimura and Matida (97). Experiments done by Mathur

(120) show that low concentrations of dieldrin and lindane have little effect on fish. In higher concentrations, both insecticies cause excitability, convulsive movements, difficulty in breathing, and finally death. Acute static bioassays were conducted with

13 pesticides to determine their comparative toxicity to fish (143).

Of the compounds tested, Thiodan, a chlorinated hydrocarbon, and

Thimet, an organic phosphorous pesticide, were the most toxic; an:1

Bayer 29493, an organic phosphorous compound, and Fermate, a carbamate, were the least toxic. Post and Garms (144) found irreversible damage to some freshwater tropical fish after severn.1 hr contact with concentrations of DDT as low as 0.05 and 0.1 mg/1,

Baytex can be tolerated for many hr at concentrations up to 3 20

Bioassays were conducted by Rao et al. (148) to establish TLmIs

of widely used chemical pesticides for Puntius puckelli, a

cyprinid.

Sreenivasan and Swaminathan (167) studied the toxicity of

paramar-50, DDVP, Metasystox, Folidol, and Dimecron to various

fish (two carp varieties, Cirrhina, 2 species of Tilapia, Danio,

Earbus and Gambusia. There was an apparent differential toxicity

to the various fish species. Neither Tenthion or Abate, organic

phosphate insecticides, were toxic to fish at a dosage rate nec-

essary to control the larvae of the midge, Glyptotendipes (190).

The toxic effects of three organophosphate insecticides, 30 percent

malathion, 50 percent thiometon, and 40 percent trichlorfon, on

female guppies, were investigated by Veger (186). Lethal time

(T ) the maximum time of exposure from which fish could be revi-red L , by removal to fresh water, was determined for the 3 pesticides.

Ruttkay (154) found that short-term exposure to sub-lethal

concentrations of dieldrin was harmless, but prolonged exposure even at low concentrations had a marked effect on the resistance

of fish to disease, and resulted finally in mortality. A population

of spot (Leiostomus xanthurus) was exposed continuously for 8

months to a sub-lethal concentration (0005 mg/1) of endrin (108).

No pathology was found in the spot after 8 months of exposure,

but a 3 wk exposure to a near-lethal concentration (0.075 mg/i)

produced pathology characterized by systemic lesions involving tilt:,

brain and spinal cord, liver, kidneys and stomach. Spot exposed 21 to 0.05 mg/1 endrin for 5 months revealed an accumulation of

78 mg/1; however, no endrin could be detected in these fish afte:.: a 13-day holding period in uncontaminated water.

Brungs and Mount (21) found the critical level of endrin in the blood of gizzard shad to be 0.10 ug/g. This level of endrin in the blood can be used for the detection of endrin-caused mortality. Production of young guppies by adults subjected to a single 2 hr exposure to dieldrin was superior to that of a control group. This was believed due to the fact that weaker fish died (79). Cairns at al. (26) also observed a similar effect on reproduction during a 14 month exposure of sub-lethal concentrations of dieldrin to guppies. During the first 2 or 3 months the exposed populations developed greater total numbers of in- dividuals than did the controls. This phenomenon has been inter- preted as an inhibition of the normal predation by the adults upcn the fry, and a slight change in the feeding behavior of the adults induced by the presence of dieldrin. Toward the end of the experiment, there was evidence that the long-term exposure had had deleterious effects.

The time to 50 percent mortality of different concentrations of C-8514 (a foramidin insecticide) and BHC to young coho salmon was determined by Velsen and Alderdice (187). Two 10-month ex- posures of malathion and butoxyethanol ester of 2,4-D under continuous-flow conditions were made to determine the effect of these pesticides on reproduction and growth of fathead minnows, 22

Pimephales promelas (129). Results show that 1/45 and 1/19 of the 96 hr TLm concentrations of malathion and butoxyethanol ester, respectively, are the highest concentrations which will not affect fhe growth and reproduction of fathead minnows during continuous exposure.

Huila et al. (130) evaluated organic pesticides for use as fish toxicants. The toxicity to carp and largemouth bass of chemicals used for mosquito and chironomid midge control was determined. Tests on the retention of vaporized lindane by 7 species of plants, fish, frogs, mice, rats, and chickens were made by

Whitacre and Ware (196). Fish (guppies and carp) were the only

test species affected in their behavior. Symptoms resulting from

the poisoning are described.

An Engineering-Science report (49) described work carried

out on the behavioral pathology of fishes resulting from exposure

to sub-lethal concentrations of toxic chemicals, including the

development and use of a special testing apparatus, known as

CARA (or Conditioned Avoidance Response Apparatus). When gold-

fish were exposed to toxaphene, behavioral aberrations were ob-

served after 264 hours at a concentration of 0.44 ug/l, but the

96-hr TLm value was 25 times greater, at 11.0 ugh. The nature

of the behavioral aberrations depended on the concentration of

the chemical and also on the period of exposure. High concen-

trations for short-time periods produced different toxic responses

than low concentrations for longer time periods. Warner et al. 23

(191) presented methods and results for rapid quantitative analysis of behavior changes in fish as a result of sub-lethal poisoning with pesticides. Analysis of changes in behavior may enable prediction of the possible consequences of sub-lethal chemical contamination of the environment,

Ferguson and Goodyear (52) found that the death rate of black bullheads whose esophagus and pharynx was closed off was not different than that of normal fishes when both were placed in endrin solutions. This indicated that endrin enters by the gills,

Gakstatter (56) studied the uptake from water by several species of freshwater fish of p,pt-DDT, dieldrin, and lindane. He also investigated the distribution of these insecticides in the tissues and their elimination rate. The metabolism of Dursban in fish was investigated by Smith at al. (164). Radioactive-labeled

Dursban was rapidly absorbed from the water by plants and soil particles, but was very slowly metabolized. Fish absorbed the compound at a much slower rate, but rapidly metabolized it.

Under natural conditions, most of the Dursban will be absorbed by the plants and soil particles thus limiting the amount taken up by the fish. Murphy (132) found that fish livers had considerably less capacity than mammalian and avian livers to inactivate the oxygen analogues of parathion and malathion, but the destruction of the anticholinesterase activity of Gutoxon by liver homogenate:3 did not differ appreciably among the three classes. The elim- 14 ination of C-labeled DDT, dieldrin, and lindane from fish 24 following a single exposure wav investigated by Gakstatter and

Weiss (57). Groups of 60 to 70 small bluegills and goldfish were exposed in polyethylene tanks to 0.03 mg/1 of the pesticides for

5 to 19 hr„ Following the expoeure, the fish were rinsed with uncontaminated water and placed in 145-liter recovery aquaria. The 14 initial lindane- C was elimthated by both species within 2 days. 14 More than 90 percent of the fmitial dieldrin- C was eliminated after 32 days of recovery. These studies also showed that labeled

DDT and dieldrin were readily transferred from contaminated fish to uncontaminated controls in the recovery aquaria.

Brungs and Bailey (20) tested the influence of suspended solids on the acute toxicity of endrin to fathead minnows. The study indicated that when endrin is introduced into natural water in an unadsorbed state its availability to fish is not greatly reduced by montmorillonite clay or Brookstone silty clay loam, only by activated carbon. Proffitt (146) found that a lack of aeration and an elevation in water temperature shortened the survival time of freshwater fishes exposed to aldrin. Studies of the effects of pesticides on goldfish previously exposed to detergents revealed that goldfish exposed to ABS are more susceptible to the toxic effects of dieldrin and DDT than are unexposed fish

(4A). Results with sodium lauryl sulfate and linear alkylate sulfonate were inconclusive. Fish held in ABS appeared to gain weight more rapidly than control fish; the larger among the ABS- exposed fish were more resistant to subsequent exposure to pesticide. 25

A study on the effects of aldrin and methoxychlor on the physical- chemical properties of water and upon Cladophora, and Daphnia, magna was made by Cabejszek (24),

Mewdesley-Thomas and Leahy (121) found that examination of tissue extracts and chromatograms of 2 size groups of pike (sox lucius) showed higher concentrations of organochlorine pesticides in the larger fish. Details are given by Holden (76) on the concentrations of dieldrin and the DDT group of pesticide residues found in salmonids from various sources. The possible biological significance of the various residue levels to fish or to predators, and the process of accumulation by fish from low aqueous concentrations to considerably higher tissue concentrations are discussee,

The bioassay of insecticide residues using crustacea, insects, and fish is discussed by Coulon (34).

Nathur (119) found that DDT and BHC solutions at various concentrations kill fish due to the absorption of these insecticides in the liver and intestine. Acetylcholinesterase (AChE) assays were conducted on 93 samples of spot and sheepshead minnows from

43 stations (77). Low enzyme activity was found in 17 samples

(18.3 percent), but 13 of these were from only 2 areas. One of these areas, in the vicinity of the Ashley River, South Carolina, receives wastes from plants producing a variety of organophosphorus compounds. Williams and Soya (197) collected distressed menhaden from the Ashley River, South Carolina and found 46.8 percent less acetylcholinesterase (AChE) activity in 26

brain homogenates as compared to menhaden collected from offshore waters. Hing (74) carried out laboratory experiments using goldfish and rainbow trout to determine whether the presence of pesticides in the water had any effect on selected blood serum enzymes in fish. It was found that the cholinesterase and transaminase enzyme systems of fish blood serum were extremely sensitive to both short-term and prolonged exposure to pesticides. The intensity of the enzyme response depended on both the period of exposure and the concentration pf pesticide. Boyle et al. (17) developed a method for infrared identification of chlorinated hydrocarbon insecticides in fish tissue. The practical limit of sensitivity needed to provide excellent infrared spectra for most common insecticides was about 1 mg/1 in the fish tissue.

Herbicides and Miscellaneous Toxicants

Bohmont (14) published a summary of the literature on the toxicity of herbicides to livestock, fish, honey bees, and wildlife. With a few exceptions, herbicides are not toxic to livestock, bees, fish, and wildlife, the exceptions being arsenic compounds, the aromatic solvents, and acrolein. Dichlone, technical hydram and treflan are also toxic to fish. Dissipation of diquat and paraquat, and their effects on aquatic weeds and fish, was studied by Yeo (201). The effects of diquat on bluegills and their food organisms was investigated by Gilderhus (61). Hematological and histopathological examinations of adult fish failed to reveal any effect of the chemical. The herbicide was toxic to Cladocera; 27

however, the populations of Cladocera built up to normal levels

after the diquat disappeared from the water. Hiltibran (73)

determined the toxicity of 15 common herbicides to the eggs of

bluegills, and the fry of bluegills, green sunfish, lake chub-

sucker, and smallmouth bass. The toxicity of 15 chemicals used

in vegetation control or disease treatment was studied by Jones

(87). Toxicity tests of three forms of silvex, three formulations

of endothal, simazine, atrazine, diquat, two forms of benzene-

hexachloride, roccal, acriflavine, malachite green, and methylene

blue were made on the fry of largemouth bass, bluegill, and

channel catfish.

Howell and King (78) reported on controlling sea lamprey,

Petromyzon marinus, with two new lampricides. Comparison of the

effects of a lampricide and of anoxia on the sea lamprey was made

by Agris (3). An electrocardiographic study of the heart of the

lamprey indicated the effects of anoxia and TFM are not the same.

Starkey and Howell (169) reported that substituted nitrosalicylan-

ilides are a new class of selectively toxic sea lamprey larvicides.

Applegate at al. (5) investigated why certain nitrophenols con-

taining halogens are significantly more toxic to larvae of sea

lampreys than to fish. It is proposed that the death of lamprey

larvae exposed to these compounds results from shock with con-

comitant circulatory and respiratory failure. Rainbow trout

appear to die, at higher concentrations of the toxicant, due to

a chemically-caused mechanical interference with respiration. 28

Penaz (140) tested the tolerance of eggs and young fish of

Salmo trutta to ammonia under laboratory conditions. None of the

concentrations (up to 50 mg/1) was found to be completely lethal

to the eggs, although receptiveness increased with the higher

ammonia concentrations. None of the exposed young trout died,

but even with concentrations of 2.10 mg/1 some irreversible damage

to heart and muscle control was observed. The effect of various

amounts of ammonia on the tissue of fish was examined by Reichen-

bach-Rlinke (150). The following significant damage occurred:

number of blood cells decreased, cells swelled, gills got lumpy,

inflammation, and hyperplasias. Recently hatched fry are the

most sensitive. Special attention was called to the fact that

tissues poor in oxygen were infiltrated by ammonia more easily

than those rich in oxygen.

Helms (70) investigated whether tadpoles could be selectively

controlled in fish hatchery ponds by the use of formalin. Results

indicated that selective control of leopard frog and young bull-

frog tadpoles may be possible. In a review on the occurrence and

study of cyanogen, Knie (100) listed the toxicity to fish of

simple and complex cyanides and thiocyanates. The toxicity to

carp of antiparasitic solutions of ammonia, chloramine, and

7 malachite green (all = ' 0.05 percent) was determined by Dobrovolny

° et al. (41). Only 0.05 percent ammonia (30-sec exposure at 12

° and 20 C) did not produce toxic effects. The lethality of the solutions was higher after hibernation. Boni (15) investigated 29

the acute toxicity and elimination of phenol injected into fish

(Carassius auratus). The effects of hydrogen sulfide on channel

catfish were studied by Bonn and Follis (16). The TLm of un-

ionized hydrogen sulfide for fry ranged from 0.8 mg/1 at pH of

6.8 to 0.53 mg/1 at pH 7.8. At pH 7.0, the TLm of this gas is

1.0 mg/1 for fingerling catfish, 103, for advanced fingerlings,

and 1.4, for adult channel catfish. Weber (193) investigated the

action of sodium nitrite on the guppy (Lebistes reticulatus). The

period of time necessary for a dose of NaNO2 to cause death in

female guppies increased with increasing calcium concentration

in the water. Death occurred earlier in older females. Babu (7)

made observations on the toxicity of the seed of Croton tiglium

on predatory and weed fishes.

Methodology

The necessity for standardized toxicity tests was dis qssed

by Breitig (18), while several other authors including Burdick (23),

Mackenthun (113), and Cairns (25) have considered present and future

techniques, problems, methods, and data treatment associated with

toxicity bioassays of waters. Definitions of toxicity, procedures

for bioassays, and new techniques for assessing toxic effects

and sub-lethal toxicological effects on living organisms are

described. Application factors suggested to derive safe field

concentrations are discussed, and the inapplicability of a

uniform dilution factor is pointed out in terms of specific

data. In their opinions, results of a bioassay are specific 30

to a particular water supply and assay conditions, and may not be

considered applicable to other waters and other conditions. Be- cause of this, toxicity values obtained from one assay series should never be applied as standards to other waters.

Problems in long-term assays are discussed by these same authors in reference to the water chemistry, fish species, desirable oxygen concentration, choice of test organism, and sim- ulation of natural conditions. Derivation of toxic concentrations by more exact methods than the Tim seems required, and it is suggested that the graphical method of Litchfield and Wilcoxon may have possibilities for this computation. It is also felt that a more refined chemical approach is needed to separate non- toxic, complexed compounds and non-active, absorbed compounds from the active toxic effects in natural waters. Bioassays with continuous-flow and constant concentration are compared with bioassays with static solutions in terms of the situations to which they are applicable. They felt some situations may be approached only by the study of natural waters in situ, assisted by constant monitoring systems now available. Finally, they suggested that while long-term assessment of pollution damage must rest on some sort of ecological study, laboratory bioassays are still required to interpret the interactions between factors.

Abram (2), in a rigorous treatment of the problem of definition and measurement of fish toxicity thresholds, gives results of four different laboratory methods of determining this 31 threshold with rainbow trout and harlequins. These methods were:

(1) long-term experiments in which the survival period was related empirically to concentrations; (2) short-term tests using a narrow range of concentrations in which only a proportion of the fish in each of a number of batches were killed; (3) comparison of survival periods in a range of constant concentrations with survival periods over the same range, but alternating with recovery periods in clean water; (4) extrapolation of the results of short-term tests by plotting the velocity of death (reciprocal of survival period) against logarithm of concentration. It was concluded that while the first method is best, the last one is simplest to use. The results of the third type of experiment suggested that, in a regularly fluctuating concentration of some poisons, if the fish survive the first high concentration period to which they are exposed, then they will react subsequently as if they were exposed to the mean concentration of the fluctuation3,

The results of investigation of the effect of 10 substances on coho eggs are presented by Yarzhombek and Grachev (200). In their studies, toxicity of a substance was expressed mathematically by the relationship between the concentrations of the dissolved substance and the time of onset of egg mortality. The relationship is hyperbolic. The maximum permissible concentration of a toxic substance in water (K) was determined by the formula

n A=(K-K0)(T-T0),0 where To and T are, respectively, the minimal and actual times of onset of mortality, K is the concentration 32 of the dissolved substance, and A and n are proportionality constants. In order to find the reasons for mortality of fish in laboratory bioassay, Hawksley (68) indicates the need for analyses with a spectroscope, pH meter, conductivity meter, and a sodium electrode. It is envisaged that in the near future a total carbon analyzer, a gas-liquid chromatograph, and/or spectro- photometer will also be required.

In meeting the challenge for more rapid and efficient conduct of bioassays, Betts et al. (12) describe a procedure for small- scale laboratory bioassays. Their procedure for rapid testing of the toxicity of a large number of effluents to fish consists of a small-scale bioassay. Equipment includes an acclimatizing tank with water circulation, temperature control, aeration, and a series of ten-gallon tanks containing a plexiglass fish-retention trough and a circulation pump. Test tanks with small fish in them are immersed in a constant-temperature water bath. This method requires less elaborate equipment, shorter testing periods, and is easier to control than full-scale continuous-flow bioassays.

New types of experimental containers include a small, open plexiglass chamber with water inlets in the ends and a common outlet in the middle described by Ganning (59). In this arrange- ment, experimental animals are offered alternative test solutions.

Brungs and Mount (21) offer a device for continuous treatment of fish in holding chambers, and De Roth (39) describes a device for aerating and circulating aquarium water without disturbing the surface water. 33

For continuously operating systems, Mount and Brungs (126)

devised a simplified diluter for maintaining a series of constant concentrations of a material in flowing water. It depends on water flows, metering cells, and venturi tubes to proportion volumes of water and toxicant to give desired concentrations.

Construction requires less than 2 days, and only readily available materials are used. An injector for mixing pesticides is also described. Another automatic dosing device for toxicity bioassay experiments is described by Stark (168).

An automatically-controlled continuous-flow apparatus designed to provide standard conditions for the bioassay of toxic substances likely to be present in sewage and industrial wastewaters is described by Calderon et al. (27). They give details of their procedure for determining the lowest concentration of a toxic substance to cause 100 percent mortality, and the highest concentration to cause no abnormal behavior in fish, using minnows and rainbow trout in tests of 6 hr duration. A new continuous- flow apparatus for measuring maximum swimming speeds of small fish is explained by MacLeod(114). The apparatus and method is considered useful in bioassays for measuring sub-lethal effects of pollutants. Fish are not required to swim at the same velocity as the water in the chamber. Rather, they swim in a current which is greater than their maximum swimming speed, which moves them

backwards in the oval channel. The average maximum swimming

speed for a group of four to six fish is calculated from current 34 velocity, circumference of the channel, and the number of laps

lost in a specified time period.

An illustrated description is given by Edeline (45) of a constant-temperature aquarium designed for fish studies in which . any selected concentration of dissolved oxygen may be maintained automatically. The oxygen content of the water is determined with a galvanic cell, having a fluorethylenepropylene polymer membrane which is said to give rapid response. Additional oxygen is supplied automatically by electrolysis to maintain the desired concentration, and the quantity is recorded continuously.

To determine the toxicity coefficient of trade wastewaters from individual sections of a plant, Donaszy (42) carried out tests with the Daphnia-guppy method. In a paper dealing with toxicity bioassays, Jackson and Brungs (84) advocate the use of newer techniques and apparatus to monitor industrial wastes, and

Lennon (107) suggests the development of a hybrid strain of fish for toxicity studies.

Temperature

Kennedy and Mihursky (93) published an excellent bibliography of 1220 references on the effects of temperature in the aquatic environment. The subject index includes aquaculture, bacteria, beneficial uses, engineering considerations, experimental techniques and apparatus, low temperatures, DO, temperature tolerance, water temperature criteria, among the 29 subject classifications. 35

An index is given which lists the different animal phyla for which data is available. Water temperature criteria necessary to protect aquatic life was discussed by Mihursky and Kennedy (122).

Effects of temperature on behavior, metabolism, and mortality of aquatic organisms are described to indicate the difficult problems involved in determining acceptable standards for a healthy fish population. Standards for temperature regulation are described for 3 ecosystems: (a) coldwater salmonid streams, (b) warm-water centrarchid environments, and (c) estuaries. Frey and Pierce (55) reported on the effects of cold water discharge on a downstream reservoir's temperature and oxygen levels. Data is also presented on the effects of the cold water discharge on a 20-mile strc.ch-. of river between the two reservoirs.

Precht et al. (145) found that in the fish, Idus idus, the frequency of opercular movement decreases continuously over a long period if the temperature to which it has been adapted is suddenly lawered. Other reactions and adaptations to the changes in the adaptation temperature were recorded for I. idus and Xiphophorus helleri. Some mechanisms underlying thermal acclimation in a freshwater fish, Etropoplus maculatus were investigated by

Parvatheswararao (138). Changes in the water, Na, K, Ca, Cl and

Mg content of the brain, liver, and muscle in warm and cold acclimated fish were determined. The possible significance of these changes in metabolic compensation to thermal stress is discussed. Strewn and Dunn (171) used response-surface diagrams 36 to compare the resistance of 10 species of marsh fishes to heat death at various salinities. The habitat of a species within the salt marsh correlates with its relative resistance to heat death.

The effect of adaptation temperature on the metabolic level of the eel, Anguilla vulgaris was determined by Jankowsky (85).

Experiments were made in order to find out why the metabolic rate of muscle in vitro does not reflect the adaptation capacity of the intact eel. Experiments by Tatarko (180), using larvae of scaly carp under laboratory conditions at various temperatures of water

(from 16° to 30°C), showed that with a rise in temperature the growth and development of carp larvae increases. DeWilde and

Houston (4D) investigated the blood 02-capacity of rainbow trout as a function of thermal acclimation. The hematological parameters of erythrocyte number, packed cell volume, hemoglobin concentrations, mean erythrocytic volume and hemoglobin content, were also investigated to evaluate the response of trout to thermal changes. Results are discussed in relation to the general problem of respiratory thermoadaptation.

A study by Shulyak (160) on the effect of temperature on development of the digestive organs of carp fry showed a definite correlation between the effect of temperature and abnormalities in the structure of the intestine. The most favorable condition for the development of the intestine was a decrease in temperature during the embryonic period followed by a rise during the postembryonic period. A definite correlation was also found between 37

the nature of the abnormalities, the growth of the fish and the

temperature at which development occurred. Smith (165) determine: the nature of temperature-induced changes in sodium-glucose interactions of the goldfish intestine. Das and Prosse (37) investigated the initial rate of protein synthesis in goldfish tissue, 14 as measured by the level and rate of incorporation of C-leucine 14 into protein. In a related paper, Des (36) again using C- leucine, investigated the synthesis of protein and RNA of subcellulnr fractions of goldfish acclimated to high and low temperatures.

Physiology and Pathology

Chavin (28) investigated physiological and morphological parameters to evaluate the responses of goldfish to small environmental changes. Preliminary results indicated that the responses of various endocrine tissues and hormone-controlled reactions to small environmental changes can be extremely rapid.

Strelltsova at al. (173) studied the influence of environmental conditions upon the physiological characteristics of several fish species. Fish acclimated in one lake had a greater tolerance to reduced dissolved oxygen and increased water temperatures than did fish from a lake with lower summer temperatures and sufficient dissolved oxygen in all water layers.

Berlin (11) observed that particulate matter was more abun- o dant in the hepatocytes of trout acclimated to 5 C than in fish acclimated at 18°C. Usdin and Hitz (184) studied the effects 38

of trace quantities of certain toxicants upon the electric discharges of some freshwater tropical fish. Many physiologically active compounds including chlorinated hydrocarbon insecticides reduced the typical frequency and amplitude of the electrical output of these fish. In the case of the insecticide chlordane, the electrical pattern returned to normal after the fish was replaced into uncontaminated water. Nikiforov (136) studied the survival, oxygen consumption and critical oxygen thresholds of young salmon in diluted artificial seawater. Fingerling and adults consume more 0 in water with a favorable salt content 2 than in fresh water. The threshold oxygen content for fry is somewhat lower in water containing up to 10 percent salt water than in fresh water. Adults have an equal threshold 02 require- ment in fresh water and in water of 10 to 15 percent salt water content.

Abou-Donia and Menzel (1) describe a rapid automatic system to assay the normal brain acetylcholinesterase of the shiner perch

(Cymatogaster) which is suitable for the demonstration of inhibition by cholinesterase-inhibiting pesticides. The optimal conditions of assay were determined and some characteristic kinetic parameters were reported such as optimal pH, temperature, and substrate concentration.

Hall and Hayton (66) studied the kinetics of the absorption of ethanol by the guppy (Lebistes reticulatus). Both time of death and time of overturn were used as criteria of pharmacological 39 effect. Guppies appeared to be suitable for drug (and toxicant) absorption studies.

Damurat (35) transferred trout, pike and roach eggs to paraffin at an early embryological stage. Embryonic develop- 0 ment occurred at temperatures from 9 to 14°C. However, only the eggs of the roach hatched.

Miscellaneous Papers

A study of the fish mortality in a small freshwater Louisiana lake was reported on by Muncy (131). The mortality was found to be associated with the development of a high population of the dinoflagellate Gymnodinium. Toxicity of the waters appeared to bs related to increased pH, length of exposure to sunlight, and concentration of the algal cells. Filtration with activated carbon removed the toxic components. Swingle (177) has discussed fish mortalities caused by phytoplankton blooms and preventive measures that may be taken. Marasas et al. (118) have found a virulent fish toxin produced by the fungus Fusarium tricinctum.

The LD of the purified toxin was 6.1 mg/kg in rainbow trout. 50 Macroscopic pathological symptoms produced by 4 to 33 mg/kg included shedding of the intestinal mucosa and severe edema.

Flavor acceptability is critical in food fishes. Pollutants have often been indicted for reducing the value of fish by lending an unacceptable taste to the flesh. Aschner et al. (6) found a muddy flavor in carp to have been caused by metabolites of Oscillatoria tenius. 40

Studies of naturally produced fish toxicants have often led to their evaluation as agents for lake rehabilitation.

Kawazu et al. (91) and (92) have reported on the toxic leaf extract of Callicarpa canadicans. They have determined its chemical structure and found its toxicity to fish to be equal that of rotenone.

Hemens (71) has recently reported on the toxicity of ammonia solutions to fish in water of varied pH. This research was directed toward explaining occasional mass mortalities of mosquito fish (Gambusia affinis) in sewage treatment lagoons. He found that lower pH values substantially reduced the toxicity of an ammonia solution. At pH 7.75 the median period of survival of mosquito fish in 120 mg/1 ammonium as nitrogen concentration was

60 minutes. At pH 6.55 no mortality occurred in 1000 minutes.

The threshold concentration was assumed to be near 1.1 mg N2/1

(as un-ionized ammonia). Male fish were significantly more sensitive than females at the same un-ionized ammonia concentration.

The production and excretion of waste nitrogen compounds in

fish has been reviewed by Makarewicz (115). He discusses the

hypothesis of the evolution of ureotelism in vertebrates. Knyazeva

(101) has reported on the introduction of carbamide (urea) into

trnut and carp. Introduction into the digestive tract, injection

into the body cavity and dissolving of carbamide in the water were tested. He found that only the introduction into the

eigestive tract prow harmless to tha 41

In a study of the effects of sediment on a trout stream,

Peters (141) found that high sediment loads and erratic stream discharge reduced the trout population of a Montana stream.

Irrigation surface return flows caused high summer surface temperatures. Survival of trout eggs was highest in stable discharge and low sediment conditions.

Guebitz (64) reported that suspended particles of several materials caused a fright reaction in rainbow trout only at concentrations causing severe turbidity of the water. Several dyes were also tested for the fright reaction. Red Napthol and Sulfur

Dye elicited severe fright behavior and were toxic at the lowest concentrations tested (1:1000 and 1:750 respectively).

Green sunfish were used by Summerfelt and Lewis (175) to test the behavioral effects of certain chemicals. Of 40 chemicals tested, 8 were found to be distinctly repellent and 1, ethanethiol, was an attractant. Alpha-chloracetophenone (ACP), or tear gas, was the most efficient repellent. It was effective at 0.5 mg/1 or one-half the median tolerance limit.

The use of dipyridylium compounds for treatment of Columnaris disease (Chondrococcus) in golden shiners was reported by Whipp

(195). Of the infected fish treated with 5.0 mg/1 dipyridylium compounds, 95 percent survived after 220 hr while only 6 percent of the untreated fish survived a like period.

Legien (106) reported a successful treatment of water to control the snail hosts of schistosome dermatitis. A 5 percent 42

suspension of Bayluscide killed all snails in 24 hr. Fish were

also killed but algae and other aquatic plants seemed unaffected.

In two papers, Kitaev (98) and (99) presents the results of

tests on the effectiveness of polychlorpinene as an ichthyocide.

He found that the polychlorpinene effect was increased with higher

water temperature. Lower water temperatures, increasing plant and

plankton, increasing pH, and a change in lake type from oligo-

trophic to eutrophic all required greater concentrations of chem-

ical to eliminate perch, roach, and pike. At 23°C and at a poly-

chlorpinene concentration of 0.15 mg/1 most fish died in one day; at 6° the fish were eliminated in 10 to 60 days.

The organophosphate insecticide, Dursban, was used to controi

moscuitos in Texas salt marshlands. Ludwig et al. (109) reported

that at effective mosquito control concentrations (25g/ha) Dursban

did not cause mortalities in associated species. Mullet and blue

crab confined in the treated areas were not affected after 21 day.

Doulding the level of application resulted in 100 percent mortality

of brown shrimp and 50 percent mortality of certain species of

minnows.

In a study of the persistence of 2,6-dichlorobenzonitrile

in aquatic environments, Valin (185) measured residues in a farm

pond after 188 days. A granular formulation added at a concentra-

tion of 0.6 mg/1 produced the highest residues in water and fish

2-wk after treatment; plants and soil showed highest levels

W. thin 1 or 2 days. With a wettable powder formulation applied 43 at 10, 20, and 40 mg/1, highest levels in water and fish were

found after 3 days. After 11 days, the concentration in water was about 2 percent of the 3-day level.

Rabinowitz and Myerson (147) have reported on the toxicity of dimethylsulfoxide (DMSO) to tropical fish. The LD50 concentration of DMSO was 1.9 percent for several species; however, other species including catfish appeared to be less sensitive. Interest- ingly, male fish were less tolerant than females of every species tested. Ball (8) reported that the 48 hr TLm of dimethyl sulfoxide to goldfish was 43 mg/l. Since acetone has a higher toxicity than this, Ball suggests DMSO as a solvent for toxicity experiments with chlorinated hydrocarbon insecticides.

*** 44

References

1. Abou-Donia, M. B„ and Menzel, D. B., "Fish Brain Cholin-

esterase: Its Inhibition by Carbamates and Automatic Assay." Comp. Biochem. Physiol., 21, 99-108 (1967).

2. Abram, F. S. H., "The Definition and Measurement of Fish Toxicity

Thresholds." Water Poll. Abs. (Brit.), 40, 1375 (1967); Proc. 3rd Intl. Conf. on Water Poll. Res., Munich, 1966,1967, 1, 75-95. 3. Agris, P. F., "Comparative Effects of a Lampricide and of

Anoxia on the Sea Lamprey." Jour. Fish. Res. Bd, Can., 24, 1819-1822 (1967).

4. Allen, G. H., and O'Brien, P., "Preliminary Experiments on the Acclimatization of Juvenile King Salmon, Oncorhynchus

tshawytscha, to Saline Water Mixed With Sewage Pond Effluent." Calif. Fish and Game, 53, 180-184 (1967).

5. Applegate, V. C., Johnson, G. G. H., and Smith, M. S., "The Relation Between Molecular Structure and Biological Activity

Among Mononitrophenols Containing Halogens." Great Lakes Fish. Comm. Tech, Rept. 11, 1-19 (1967); Biol. Abs., 48, 85850 (1967).

6. Aschner, M., Laventer, C„ and Chorin-Kirsch, I., "Off Flavour

in Carp from Fish Ponds in the Coastal Plain and the Galil." Bamidgeh, 19, 23-25 (1967).

7. Babu, N., "Observations on the Toxicity of the Seed of Croton

tiglium. Linn. on Predatory and Weed Fishes." Sci. Cult., 31, 308-310 (1965); Biol. Abs., 48, 11125 (1967). 45

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