Comparison of acute toxicity of ketoprofen to juvenile and embryonic stages of Danio rerio.

OBJECTIVES: Ketoprofen is a common human medicine from a class of non-steroidal anti-inflammatory drugs (NSAIDs), which is provably detected in surface waters in concentrations ordinarily in µg.L-1. The aim of this study was to compare the acute toxicity of ketoprofen to embryonic and juvenile stages of aquarium fish - zebrafish (Danio rerio). METHODS: Tests were performed according to the methods of the Organisation for Economic Co-operation and Development (OECD) No. 203 (Fish, acute toxicity test) and OECD No. 212 (Fish, short-term toxicity test on embryo and sac-fry stages). RESULTS: The results showed (mean ± SD) LC50 value of ketoprofen to be 632.30 ± 10.10 mg.L-1 in juvenile zebrafish and 6.44 ± 2.22 mg.L-1 in embryonic stages of zebrafish. The results revealed statistically significantly higher sensitivity (p<0.01) of the embryonic stages of zebrafish to ketoprofen compared to its juveniles. The susceptibility of embryos depends on many factors, especially yet improperly developed enzymatic system in embryos, different ways of the absorption of the substance into the organism or differences in metabolism pathways. CONCLUSIONS: The acute toxicity of ketoprofen for juvenile stages of zebrafish is low, but the substance seems to be toxic for embryonic stages.

Effects of subchronic exposure to simazine on zebrafish (Danio rerio).

OBJECTIVES: Simazine is a triazine herbicide which has been used for a long period in agriculture and in the aquatic environment for control of weeds and algae. The aim of this study was to investigate the effects of subchronic exposure to simazine on growth and the development of histopathological changes in selected organs (gills, kidney, liver) in Danio rerio. METHODS: Juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the environmental simazine concentration commonly detected in Czech rivers (0.06 µg.L-1) and a range of sublethal concentrations of simazine (0.6, 6.0 and 60.0 µg.L-1) for 28 days. RESULTS: There were no significant differences (p<0.05) between the specific growth rates (r) of the test groups and those of both control groups. Histopathological examination revealed pathological changes in fish exposed to a simazine concentration of 60.0 µg.L-1. The values of NOEC and LOEC of simazine were 6.0 µg.L-1 and 60.0 µg.L-1. CONCLUSIONS: The environmental concentration of simazine in Czech rivers did not have any effects on the growth and development of histopathological changes in D. rerio.

Comparison of diazinon toxicity to embryos of Xenopus laevis and Danio rerio; degradation of diazinon in water.

The aim of this study was to determine the toxic effect of diazinon (organophosphate insecticide) to embryos of Xenopus laevis and Danio rerio. The 96-h LC50 values showed higher toxicity of diazinon for X. leavis in standard solution (9.84 mg/L) compared to the pond water (12.64 mg/L). Teratogenic index for diazinon was 1.3 and 1.6, respectively. The 96-h LC50 diazinon values demonstrated similar sensitivity of embryos D. rerio (8.21-9.34 mg/L) and X. laevis in standard test solutions. Our results reflect that direct application of diazinon into the water can be associated with significant risks to aquatic organisms.

Effect of nitrite on early-life stages of common carp (Cyprinus carpio L.).

A one-month chronic exposure of common carp larvae and embryos to nitrite revealed significant (p < 0.01) differences in total accumulated mortality in fish exposed to 33, 67, and 330 mg/L NO(2)(-) compared with controls. At the highest concentration, all fish died within 8 d of exposure. On the basis of accumulated mortality in the experimental groups, lethal concentrations of nitrite were estimated at 29 d LC50 = 88 mg/L NO(2)(-); lowest-observed-effect concentration (LOEC) = 28 mg/L NO(2)(-); and no-observed-effect concentration (NOEC) = 7 mg/L NO(2)(-). Fulton's condition factor values were significantly lower in fish from all experimental groups compared with controls. By day 12, fish exposed to 33 and 67 mg/L NO(2)(-) had significantly lower mass and total length compared with controls. No significant negative effects of nitrite at the concentrations tested (0.7-330 mg/L NO(2)(-), at 10 mg/L Cl(-)) on hatching or embryo viability were demonstrated, but significant differences in early ontogeny among groups were noted. Fish from all the concentrations showed a dose-related delay in development compared with the controls. Lordosis, kyphosis, scoliosis, and body shortening were observed at all concentrations and in controls, as was yolk sac deformation and edema, eye deformation, and cardiac edema. The incidence of these malformations was positively correlated with nitrite concentration. Histopathology revealed epidermal spongiosis; edema and hyperplasia of the gill epithelium, including hypertrophy and hyperplasia of eosinophilic granular cells (chloride cells); and interstitial edema of skeletal muscle in fish exposed to 67 mg/L NO(2)(-). Similar, but milder, changes were observed at lower nitrite concentrations.

Polyaluminium chloride (PAX-18) - acute toxicity and toxicity for early development stages of common carp (Cyprinus carpio).

OBJECTIVES: The aim of this study is to assess the toxicity of PAX-18 in different developmental stages of common carp (Cyprinus carpio). The preparation PAX-18, with its active ingredient polyaluminium chloride (9% of Al), is a coagulation agent that is used mainly to precipitate phosphates, to prevent surface water eutrophication and incidences of cyanobacteria. It is applied to the water environment and thus could present a potential risk to fish. DESIGN: The toxicity tests were performed on common carp according to OECD (203, 210) methodologies. The acute toxic effect was evaluated for juveniles and the early development stage effect was observed in embryo-larval toxicity tests. The results of the toxicity tests (the number of dead individuals at particular test concentrations) were subjected to a probit analysis using an EKO-TOX 5.2 programme to determine the LC50 values of PAX-18. RESULTS: Acute toxicity value expressed as 96hLC50 was 753.1 +/- 24.3 mg.l-1 (67.8 mg.l-1 Al). Effect on early development stage expressed as the no observed effect concentration was 10 mg.l-1 (0.9 mg.l-1 Al), the lowest observed effect concentration was 50 mg.l-1 (4.5 mg.l-1 Al). No significant effects of the preparation PAX-18 in concentrations of 50 mg.l-1 of PAX-18 and lower were found on hatching, length and weight parameters, morphology and histopathology. CONCLUSION: The lethal concentration of PAX-18 found in acute toxicity tests on common carp was 7-14 times higher than the concentration which is usually applied to water (5-10 mg.l-1 Al). Moreover, fish in eutrophicated water sources are exposed to PAX-18 concentrations corresponding with the lowest observed effect concentration only for a short time, therefore the effect on them can be considered as minimal.

Terbutryn toxicity to Danio rerio: effects of subchronic exposure on fish growth.

OBJECTIVES: The aim of this study was to investigate effects of subchronic exposure to sublethal levels of terbutryn on growth and on histopathological changes in Danio rerio. DESIGN: The acute toxicity tests were performed on the juvenile stage of Danio rerio according to OECD No. 203 Fish, Acute Toxicity Test. The juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the terbutryn environmental concentration commonly detected in the Czech rivers (0.02 microg L-1) and the range of sublethal concentrations of terbutryn (0.06, 0.2, 0.6 and 2 mg L-1) for 28 days. RESULTS: The 96hLC50 terbutryn mean value for the juvenile stage of D. rerio was 5.71 +/- 0.46 mg L-1. A significant decrease (p < 0.01) in the fish growth caused by terbutryn was observed in the concentration of 0.6 mg L-1. The value of NOEC was 0.2 mg L-1 and LOEC was 0.6 mg L-1 of terbutryn. We found the damage to tubular system of kidneys in the concentration of 0.6 mg L-1 of terbutryn. CONCLUSION: The environmental concentration of terbutryn in the Czech rivers did not have any effects on growth and on histopathological changes in D. rerio and this concentration is lower than determined LOEC and NOEC values of terbutryn.

Comparison of the sensitivity of different fish species to medical substances.

OBJECTIVES: Formaldehyde, sodium chloride and potassium permanganate belong to the commonly used substances for fish treatment. The aim is to define and compare their acute toxicity and therapeutic index between two fish species - Danio rerio and Poecilia reticulata. DESIGN: To determine acute toxicity of these compounds, the semistatic method was implemented in compliance with the OECD No. 203 (Fish acute toxicity test). In each test series, 4 acute toxicity tests were performed. The results were subjected to the probit analysis to determine the 96hLC50 values. Therapeutic index (TI) was calculated with respect to short-term LC50 and effective therapeutic concentrations commonly used for fish treatment. RESULTS: The mean 96hLC50 values for sodium chloride were 21.69 +/- 0.92 g l-1 (TI = 1.4) for P. reticulata and 10.39 +/- 0.12 g l-1 (TI = 1.0) for D. rerio. The acute toxicity of formaldehyde for D. rerio expressed as 96LC50 was 0.12 +/- 0.003 ml l-1 (TI = 5.24) and for P. reticulata 0.1 +/- 0.003 ml l-1 (TI = 4.9). The acute toxicity of potassium permanganate for D. rerio expressed as 96LC50 was 1.25 +/- 0.15 mg l-1(TI = 1.5) and for P. reticulata 1.43 +/- 0.05 mg l-1 (TI = 2). CONCLUSION: P. reticulata showed significantly higher (p < 0.05) tolerance to sodium chloride than D. rerio, whereas D. rerio showed significantly higher (p < 0.05) tolerance to formaldehyde than P. reticulata. The acute toxicity of potassium permanganate was comparable for both fish species. Calculated therapeutic indexes of all tested substances were low; therefore it is important to conduct preliminary tolerance tests before application of treatment baths.

Leeches as Sensor-bioindicators of River Contamination by PCBs.

The aim of the study was to evaluate the use of leeches of the genus Erpobdella as a means of assessing polychlorinated biphenyl contamination of watercourses. The River Skalice, heavily contaminated with PCBs, was selected as a model. The source of contamination was a road gravel processing factory in Rozmitál pod Tremsínem from which an estimated 1 metric ton of PCBs leaked in 1986. Levels of PCB were measured in leeches collected between 1992 to 2003 from 11 sites covering about 50 km of the river (the first sampling site upstream to the source of contamination and 10 sites downstream). The PCB indicator congeners IUPA no. 28, 52, 101, 118, 138, 153, and 180 were measured. Levels were highest at the four sampling sites nearest the source of pollution. The highest values of PCB congeners were found in 1992. PCB content decreased from 1992 to 2003 and with distance from the source. The study indicated that leeches of the genus Erpobdella are a suitable bioindicator of contamination in the surface layer of river sediments.

Comparison of acute toxicity of 2-phenoxyethanol and clove oil to juvenile and embryonic stages of Danio rerio.

OBJECTIVES: Anaesthetics are used in aquaculture to prevent stress and mechanical damage to fish during handling or the treatment of fish in breeding, blood sampling and other veterinary interventions. Clove oil and 2-phenoxyethanol are used in the Czech Republic in a water bath for the short-term immobilization of the fish. DESIGN: Acute toxicity tests were performed on aquarium fish Danio rerio, which is considered to be one of the model organisms most commonly used in toxicity testing. The semi-static method according to OECD No. 203 (Fish acute toxicity test) was used for testing juvenile fish. Embryo toxicity tests were performed in zebrafish embryos (D. rerio) in compliance with the OECD No. 212 methodology (Fish, short-term toxicity test on embryo and sac-fry stages). The results obtained (the number of dead individuals at particular test concentrations) were subjected to a probit analysis using the EKO-TOX 5.2 programme in order to determine LC50 clove oil and 2-phenoxyethanol values. The statistical significance of the difference between LC50 values in juvenile and embryonic stages of D. rerio was tested using the Mann-Whitney non-parametric test implemented in the Unistat 5.1 programme. RESULTS: The LC50 clove oil mean value was 18.8 +/- 5.52 mg.L-1 in juvenile D. rerio, and 15.64 +/- 3.30 mg.L-1 in embryonic stages of D. rerio. The LC50 2-phenoxyethanol mean value was 338.22 +/- 15.22 mg.L-1 in juvenile D. rerio, whereas in embryonic stages of D. rerio it was 486.35 +/- 25.53 mg.L-1. CONCLUSIONS: The study proved statistically significantly higher (p<0.01) sensitivity in juvenile fish to 2-phenoxyethanol compared to the embryonic stages. Acute toxicity values of clove oil for juvenile and embryonic stages were comparable.

Comparison of the sensitivity of Danio rerio and Poecilia reticulata to silver nitrate in short-term tests.

The aim of this study is to assess the acute toxicity of silver nitrate in adult zebra fish and adult guppies and to compare the sensitivity of these species to this compound. Silver is a naturally occurring element in our environment and it combines with other elements such as sulfide, chloride, and nitrate. Silver, in the form of silver nitrate, is one of the most toxic metals affecting freshwater fish. Industry, particularly photographical and electrotechnical, is the major contributor of silver that is released into the environment. Tests of acute toxicity were performed on the most common species of aquarium fish, Danio rerio and Poecilia reticulata. Both zebra fish and guppies were exposed to progressive concentrations of silver nitrate; a semi-static method according to OECD 203 was used. In each test series, 6 tests of acute toxicity were conducted, with 10 fish used for each separate concentration and for the control group. The results (number of fish deaths in the individual test concentrations) were subjected to probit analysis (EKO-TOX 5.1 software) to determine the 96hLC(50) AgNO(3) values. The 96hLC(50) AgNO(3) value for the zebra fish was (mean±SEM) 15±0.52 µg/l and for the guppies was (mean±SEM) 17.14±5.43 µg/l. We didn't find any statistically significant difference between the sensitivity of zebra fish and guppies. The results reported in this study are in agreement with LC(50) values published in peer-reviewed literature, and conclude that AgNO(3) is one of the most toxic compounds known to fishery.