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«: AGROCHEMICALS: FATE IN FOOD AND THE ENVIRONMENT PROCEEDINGS OF A SYMPOSIUM, ROME, 7 - 1 1 JUNE 1982 JOINTLY ORGANIZED BY IAEA AND FAO l^J I N T E R ...»

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Fish play an important role in the assessment of the fate and effect of pesticides in the aquatic environment, both on intentional and unintentional release. The roles of fish toxicity and human consumption are discussed with respect to uptake, metabolism and excretion, and including residues. In this context a great deal of work has been done with labelled pesticides which has had to be evaluated comparatively with other organisms. Accumulation, as a factor of pesticide mobility and an indicator of toxicity, has been very intensively studied in fish. As the latter are model ecological organisms for accumulation, many parallel investigations have been carried out with labelled and non-labelled material. Owing to the generally low metabolic rate detected, accumulation in fish is largely due to the parent compound in the case of persistent pesticides. The methods used ranged from simple laboratory tests to experiments in the open environment.

INTRODUCTION

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HAZARD-RANKING I n v e s t i g a t i n g p e s t i c i d e s in fish c o n s i s t s of several a p p r o a c h e s and a variety of experimental methods. T h e s e also include investigations where fish are but one parameter in a set of investigative factors.

<

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FIG.3. DDT concentrations in various Perch tissues (о muscle; • gills; • liver; • mesenteric adipose) as a function of time after labelling the pond. From Ref. [4].

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adipose, c o m p a r i s o n s could not be made. For both species, the lowest levels were found in the m u s c l e tissue. T h e highest c o n c e n t r a t i o n (16.1 ppm) found in the P e r c h liver was m e a s u r e d in the one-day samp l e. It declined fairly rapidly thereafter, showing a fast tissue response to the DDT added to the water. T h e highest c o n c e n t r a t i o n in the liver of the Crucian carp (0.39 ppm) was measured after four days. T h i s is below the lowest value of the Perch liver (0.52 p p m ), and was measured at the end of the test p e r i o d.

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FIG.4. Relationship between the unextractable radioactivity in Mosquito fish (Gambusia affinis) at the average change in concentration of radioactivity from day 3 to day 9 (ppm 9/3j and day 10 to day 16 ¡ppm 16/10).

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FIG.5. Elimination of 1AC-4-CB residues during early development of Rainbow trout. Each point and associated vertical line represents the mean ± SE (n = 15). The tl¡2 of 231 days was calculated from means for days 1 - 42; tl¡2ofl4.6 days was calculated from means for days 49- 105. The time course of the early development in this study is given at the top of the figure and the duration of 4-CB exposure at the lower left. From Ref. [9].

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L e e at al. [15] studied uptake, metabolism and elimination of - L 4 C-naphthalene and 4-benzopyrene by marine fish. T h e s e chemicals are mentioned although they are not p e s t i c i d e s but occur globally.

T h r e e species of marine fish (Mudsucker or Sand goby, Gillichtys mirabilis; Sculpin, Oligocottus maculosus; Sand dap, C i t h a r i c h t h y s stiqmaeus) were exposed to the two chemicals for various periods of time. W i t h i n m i n u t e s all three fish species rapidly absorbed the hydrocarbon through the gills. R a d i o a c tivity build-up was mainly in the liver where metabolism took place. E l i m i n a t i o n was via the bile.

N a p h t h a l e n e resorption was more efficient than uptake of benzpyrene. Based on the radioactivity data in the liver, there was already a steady-state of c o n c e n t r a t i o n after a few hours of exposure. Major m e t a b o l i t e s were identified as dihydroxynaphthalene and d i h y d r o d i h y d r o x y b e n z p y r e n e. T h e gall bladder was the major organ for storing chemicals and metabolites, consequently urine was the most important excretion route. After termination of exposure the radioactivity in various tissues was reduced tenfold within 24 hours. E l i m i n a t i o n of naphthalene was in total greater than that of benzpyrene. A g a i n, in this case there are the same detoxification mechanisms prevailing as already m e n t i o n e d, namely conjugation with sulphate and glucosiduronic acid.

T h e oncogenetic effects of the so-called carcinogen benzo(a)pyrene (BAP) on three species of larval Flat fish were investigated by H o s e et al. [16], using c o n c e n t r a t i o n s from 0.10 to 4.2 ppb, which were comparable to levels found in polluted harbours. It is not usual that specific toxicity investigations are done using the labelled material, but in this case the l ^ c - l a b e l l e d BAP was used to facilitate analyses, e.g. for taking autoradiographs. B A P - t r e a t e d sand sole (Psettichthys m e l a n o stichus) eggs showed a decline in hatching success and a higher incidence of anomalies than did control eggs. Flathead sole (Hippoglossoides elassodon) eggs exposed to a single dose of a water-soluble B A P bovine serum albumin complex demonstrated evidence of toxic injury with pycnotic nuclei present in the integument and, more c o m m o n l y, in ocular and neural tissues. An increased incidence of morphological anomalies in E n g l i s h sole (Parophyrs vetulus) eggs and larvae exposed to BAP was not detected [16].





158 F R E I T A G and K L E I N

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FIG.11. Radio-thin-layer chromatograms of 3sS-C12-AES(3), 3sS-C12-AES(5) and their metabolites on silica gel developed in l-butanol: acetic acid: water (4:1:1) in Carp exposed

to 0.56-0.63 mg/L solutions A-bile extract of fish exposed to 3SS-C12-AES(3):

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FIG. 13(a). Effect of varying the scale of the system on the level of bioaccumulation in fish Common minnow /Timephales promelasj. The volume of water, amounts of DDT and sand were proportionally varied, except that the size of fish was kept constant, (b) Effect of varying the scale of the system including the size of fish on the level of DDT accumulation, (c) Variation of the water volumes affecting the level of bioaccumulation. In this experiment, the amount of DDT and sand and the size of fish were not changed proportionally.

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FIG.1.4(a). Effect of variation offish sizes on the level of bioaccumulation of DDT in Common minnow fPimephales promelas^ under standard model conditions, (b) Effect of varying the depth of the system, at a constant volume of water, on the level of bioaccumulation of DDT in fish, (c) Effect of changes in the amount of applied DDT on the level of bioaccumulation of DDT (the amount of sand was also proportionally changed), (d) Effect of changes in ambient temperature on the level of DDT accumulation in fish.

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FIG.15. Accumulation factor of TCB and PCP in Golden Orfe fLeucisens idus melanolusj and Zebra fish fBrachydanio rerio) ( temperature).

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FIG.16. Accumulation factor of TCB and PCP in Golden Orfe fLeucisens idus melanolus^ and Zebra fish fBrachydanio rerioj by different pH-values of the water.

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[1] KORTE, F., FREITAG, D., GEYER, H., KLEIN, W „ KRAUS, A., LAHANIATIS, E., Ecotoxicological profile analysis: a concept for establishing ecotoxicologic priority lists for chemicals, Chemosphere 7 1 (1978) 79.

[2] FREITAG, D „ GEYER, H., KLEIN, W „ KRAUS, A., LAHANIATIS, E „ KORTE, F., An approach for comparative screening of the environmental behaviour of chemicals, Ecotoxicol. Environ. Saf. 3 (1979) 144.

[3] FREITAG, D., GEYER, H., KRAUS, A., VISWANATHAN, R., KOTZIAS, D., ATTAR, A., KLEIN, W., KORTE, F., Ecotoxicological profile analysis. VII. Screening chemicals for their environmental behaviour by comparative evaluation, Ecotoxicol.

Environ. Saf. 6 ( 1 9 8 2 ) 60.

[4] SALONEN, L., VAAJAKORPI, H.A., "Bioaccumulation of 14 C-DDT in a small pond", Comparative Studies of Food and Environmental Contamination (Proc. Symp. Otaniemi, 1973), IAEA, Vienna (1974) 201.

[5] GASITH, A., PERRY, A.S., "Fate of parathion in a fish pond ecosystem and its impact on food-chain organiams. Part of an integrated study of substrate-biological species interaction in an aquatic ecosystem", Agrochemical Residue-Biota Interactions in Soil and Aquatic Ecosystems (Proc. Panel Vienna, 1978), IAEA, Vienna ( 1 9 8 0 ) 125.

[6] SANBORN, J.R., CHILDERS, W.F., METCALF, R.L., Uptake of three polychlorinated biophenyls, DDT, and DDE by the Green Sunfish, Lepomis cyanellus Raf., Bull. Environ.

Contam. Toxicol. 13 2 (1975) 209.

166 FREITAG and KLEIN [7] METCALF, R.L., " A laboratory model ecosystem for evaluating the chemical and biological behaviour of radiolabeled micropollutants", Comparative Studies of Food and Environmental Contamination (Proc. Symp. Otaniemi, 1973), IAEA, Vienna (1974) 49.

[8] HERBST, E., WEISGERBER, I., KLEIN, W., KORTE, F., Beitráge zur okologischen Chemie CXVIII, Bilanz, Bioakkumulierung und Umwandlung von 2,2-Dichlorbiphenyl-14C in Goldfischen, Chemosphere 2 (1976) 127.

[9] GUINEY, P.D., LECH, J.J., PETERSON, R.E., Distribution and elimination of a polychlorinated biphenyl during early life stages of Rainbow trout (Salmo gairdneri), Toxicol. Appl. Pharmacol. 53 (1980) 521.

[10] MELANCON, M.J. Jr., LECH, J.J., Isolation and identification of a polar metabolite of tetrachlorobiphenyl from bile of Rainbow trout exposed to 14C-tetrachlorobiphenyl, Bull. Environ. Contam. Toxicol. 15 2 (1976) 181.

[11] PRITCHARD, J.B., GUARINO, A.M., KINTER, W.B., Environ. Health Perspect. 4 (1973)45.

[12] SANBORN, J.R., CHILDERS, W.F., HANSEN, L.G., Uptake and elimination of (14C)hexachlorobenzene (HCB) by the Green Sunfish, Lepo mis cyanellus Raf., after feeding contaminated food, J. Agrie. Food Chem. 25 3 (1977) 551.

[13] KOBAYASHI, K., "Metabolism of pentachlorophenol in fishes", Environmental Science Research, Pentachlorophenol: Chemistry, Pharmacology and Environmental Toxicology (RAO, K.R., Ed.) Vol.12, Plenum Press, New York (1978).

[14] TULP, M.Th., H A Y A, К., CARSON, W.G., ZITKO, V., HUTZINGER, O., Effect of salinity on uptake of 14 C-2,2', 4,5,5'-pentachlorobiphenyl by juvenile Atlantic salmon, Chemosphere 4 (1979) 243.

[15] LEE, R.F., SAUERHEBER, R., DOBBS, G.H., Uptake, metabolism and discharge of polycyclic aromatic hydrocarbons by marine fish, Mar. Biol. 17 (1972) 201.

[16] HOSE, J.E., HANNAH, J.B., DIJULIO, D., LANDOLT, M.L., MILLER, B.S., IWAOKA, W.T., FELTON, S.P., Effects of benzo(a)pyrene on early development of Flatfish, Arch. Environ. Contam. Toxicol. 11 (1982) 167.

[17] KIKUSHI, M., WAKABAYASHI, M., KOJIMA, H., YOSHIDA, T., Bioaccumulation sodium alkylpoly(oxyethylene) sulfates in Carp (Cyprinus carpió), profiles of 35S-labelled Water Res. 1 4 ( 1 9 8 0 ) 1541.

[18] LECH, J.J., GLICKMAN, A.H., STATHAM, C.N., "Studies on the uptake, disposition and metabolism of pentachlorophenol and pentachloroanisole in Rainbow trout (Salmo gairdneri)", Environmental Science Research, Pentachlorophenol: Chemistry, Pharmacology and Environmental Toxicology (RAO, K.R., Ed.) Vol.12, Plenum Press, New York (1978).

[19] MATSUMURA, F., "Absorption, accumulation, and elimination of pesticides by aquatic organisms", Environmental Science Research, Pesticides in Aquatic Environments (KHAN, M.A.Q., Ed.) Vol.10, Plenum Press, New York (1977).

[20] BUDE, A., Bestimmung des quantitativen Einflusses von biologischen, physikalischen und chemischen Faktoren auf die Bioakkumulation in Fischen am Beispiel von 2,5,4'-Trichlorbiphenyl und Pentachlorphenol, Dissertation TU München-Weihenstephan, 1981.

IAEA-SM-263/31

APPLICATION OF A RADIOMETRIC ENZYMIC

METHOD FOR MONITORING ORGANOPHOSPHOROUS

AND CARBAMATE INSECTICIDE RESIDUES

IN WATER OF THE RIVER DANUBE*

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APPLICATION OF A RADIOMETRIC ENZYMIC METHOD FOR MONITORING ORGANOPHOSPHOROUS AND CARBAMATE INSECTICIDE RESIDUES IN WATER OF THE RIVER

DANUBE.

The necessity of a monitoring method that is sensitive to the anti-cholinesteratic quality of pesticides is obvious. For this purpose we applied a radiometric version of the enzymatic determination technique. Acetylcholine labelled with tritium in the acetyl moiety was used as substrate. The method is suitable for detection of the ppm level of organophosphorous or carbamate insecticide residues in water. From 1978 onwards water of the river Danube was regularly monitored by means of the radiometric enzymic method. In the minority of cases significant levels of residues were detected. Samples of extreme values were extracted and analysed by gas chromatography. During the monitoring programme it was recognized that water samples possess an acetylcholine hydrolysing ability. This parameter was also detected and plotted. The method was also used in laboratory model experiments for investigating degradation of pesticides in various water samples.

* This work was supported, and co-ordinated by the IAEA under Research Contract No. 1793/R5/RB. The abstract'only is published, since it is intended that the full paper will appear in the IAEA-TECDOC Series (unpriced publication).

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INFLUENCE OF THE ORGANOPHOSPHOROUS INSECTICIDES ACEPHATE A N D

PARATHION ON THE HETEROTROPHIC BACTERIA OF TWO FRESHWATER

ECOSYSTEMS.



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