Investigation of acute toxicity of chlorpyrifos-methyl on Nile tilapia (Oreochromis niloticus L.) larvae

Chlorpyrifos-methyl, a wide-spectrum organophosphorus insecticide and potential toxic pollutant contaminating aquatic ecosystems, was investigated for acute toxicity. Larvae of the freshwater fish Nile tilapia (Oreochromis niloticus L.) were selected for the bioassay experiments. The experiments were repeated three times and the 96 h LC50 was determined for the larvae. The static test method for assessing acute toxicity was used. Water temperature was maintained at 25+/-1 degrees C. In addition, behavioral changes at each chlorpyrifos-methyl concentration were observed for the individual fish. Data obtained from the chlorpyrifos-methyl acute toxicity tests were evaluated using Finney's probit analysis statistical method. The 96 h LC50 value for Nile tilapia larvae was calculated to be 1.57 mg/l.     

Investigation of acute toxicity of permethrin on guppies Poecilia reticulata

Permethrin, a synthetic pyrethroid pesticide and potential toxic pollutant contaminating aquatic ecosystems, was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) were selected for the bioassay experiments. The experiments were repeated 3 times and the 48-h LC(50) was determined for the guppies. The static test method of acute toxicity test was used. Water temperature was regulated at 20+/-1 degrees C. In addition, behavioral changes at each permethrin concentration were observed for the individual fish. Data obtained from the permethrin acute toxicity tests were evaluated using the probit analysis statistical method. The 48-h LC(50) value for guppy was estimated as 245.7 microg/l. Values in the range of 0.05-97.0 microg/l have been reported for various other fish species.     

Investigation of acute toxicity of beta-cypermethrin on guppies Poecilia reticulata

Beta-cypermethrin, a synthetic pyrethroid pesticide and potential toxic pollutant, contaminating aquatic ecosystems was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) was selected for the bioassay experiments. The experiments were repeated three times and the 48-h LC50 was determined for the guppies. The static test method of acute toxicity test was used. Water temperature was regulated at 22 +/- 1 degrees C. In addition, behavioral changes at each beta-cypermethrin concentration were observed for the individual fish. Data obtained from the beta-cypermethrin acute toxicity tests were evaluated using the probit analysis statistical method. The 48-h LC50 value for guppy was estimated as 21.4 microg/l.     

Water quality and fish size affect toxicity of endosulfan, an organochlorine pesticide, to rainbow trout

The acute toxicity of endosulfan in juvenile rainbow trout (Oncorhynchus mykiss, 10.61+/-1.69 g) was evaluated in glass aquaria under static conditions. Nominal concentrations of endosulfan in the toxicity test ranged from 1.3 microg l(-1) to 29 microg l(-1). The concentrations of endosulfan that killed 50% of the rainbow trout within 24-h (24-h LC50), 48-h LC50, 72-h LC50, and 96-h LC50 were 19.78, 8.89, 5.28, and 1.75 microg l(-1), respectively. None of the unexposed control fish died, and the first fish died 4 h after exposure to 26.3 microg l(-1) of endosulfan. Survival of fish was significantly increased with increasing fish size and decreased with decreased fish size at the same temperature (p<0.001). Temperature also had a significant effect on survival of fish. Alkalinity at levels above 20 mg l(-1) as CaCO3 significantly increased survival of fish at 19.78 microg l(-1) of endosulfan. Increasing alkalinity from 20 mg l(-1) as CaCO3 to 42 or higher concentrations tested in this study (121 mg l(-1) as CaCO3) significantly increased survival of fish (p<0.01). Total hardness ranging from 55 mg l(-1) as CaCO3 to 126 mg l(-1) as CaCO3 did not affect survival of fish exposed to endosulfan. Endosulfan toxicity was found to be irreversible when fish were exposed to minimum concentrations of endosulfan tested. Histologically, fish gills had lamellar edema, separation of epithelium from lamellae, lamellar fusion, and swelling of the epithelial cells. Melanomacrophage centers were scattered throughout the trunk kidney, head kidney, and spleen. The liver of endosulfan-exposed fish had severe focal necrosis. None of these lesions were seen in unexposed control fish. Results indicate that alkalinity, temperature, and fish size affect endosulfan toxicity of rainbow trout.     

Arsenate and perchlorate toxicity, growth effects, and thyroid histopathology in hypothyroid zebrafish Danio rerio

Exposure to perchlorate or other thyrotoxic compounds can cause hypothyroidism in most vertebrates, and this may affect levels of endogenous antioxidants and cause oxidative stress. Arsenic also induces oxidative stress in animals by modifying the antioxidant capacity and may alter the thyroid homeostasis. Therefore, hypothyroidism may affect the toxicity of arsenate. In order to test this hypothesis, zebrafish (Danio rerio) were made hypothyroid by exposure to perchlorate, and toxicity of arsenate in hypothyroid and euthyroid fish was compared. The endpoints were LC50 and thyroid histopathology. Additionally, the recovery of thyroid histopathological indices after cessation of perchlorate exposure was determined. The current study showed that 96 h LC50 of perchlorate anion and arsenate ion to juveniles fish (37 day post-fertilization) were 2532 and 56 mg l(-1), respectively. In addition, hypothyroid fish were more sensitive to arsenate, with a 96 h LC50 of 43 mg l(-1). Growth rates were also significantly retarded by perchlorate exposure. After cessation of perchlorate exposure, there was recovery of thyroid histopathology in terms of epithelial cell height, but not colloid area or growth rate. In conclusion, perchlorate enhances arsenate toxicity to juvenile zebrafish, and the rate of thyroid recovery after cessation of perchlorate exposure depends on the endpoints examined.     

Investigation of acute toxicity of fenitrothion on peppered corydoras (Corydoras paleatus) (Jenyns, 1842)

Fenitrothion, as an organophosphothionate insecticide, is a contact insecticide and selective acaricide, also used as a vector control agent for malaria in public health programs. A 96 h LC50 value of fenitrothion, a potential toxic pollutant contaminating aquatic ecosystems, was determined on the adult peppered corydoras (Corydoras paleatus). The experiments were repeated three times. The static test method of acute toxicity test was used. Water temperature was regulated at 23 +/- 1 degrees C. In addition, behavioral changes at each fenitrothion concentration were observed for the individual fish. Data obtained from acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The 96 h LC50 value for peppered corydoras was estimated as 3.51 mg/l.     

Effects of chlorpyrifos on reproductive performances of guppy (Poecilia reticulata)

Guppy (Poecilia reticulata) was selected to investigate the effects of chlorpyrifos on reproductive performances. Male and female guppy with proven fertility were selected from our own colony and the groups of fish (n=72/group) were exposed to pre-determined chlorpyrifos concentrations (0.002 microg/l, 2 microg/l) based on the 96-h LC50 for guppy. Mating behavior of males was recorded on the 2nd day of exposure. Offspring were counted and survival recorded on the 14th day. Gonopodial thrusts (8/15 min) in 0.002 microg/l and (4/15 min) in 2 microg/l were significantly different from the control group (11/15 min). Similarly, live birth reduced significantly to 8/female in 2 microg/l compared to 27/female in the control group. Survival of offspring after 14 days was reduced to 47% in the 2 microg/l group compared to 94% of survival in the control. Our findings demonstrate that low soluble concentrations of chlorpyrifos affect mating behavior, number of offspring and offspring survival of guppy.     

Comparative study on the susceptibility of freshwater species to copper-based pesticides

Copper compounds have been intentionally introduced into water bodies as aquatic plant herbicides, algicides and molluscicides. Copper-based fertilizers and fungicides have been widely used in agriculture as well. Despite the fact that copper is an essential element for all biota, elevated concentrations of this metal have been shown to affect a variety of aquatic organisms. Nonetheless, comparative studies on the susceptibility of different freshwater species to copper compounds have seldom been performed. This study was conducted to compare toxicity of copper-based pesticides (copper oxychloride, cuprous oxide and copper sulfate) to different freshwater target (Raphidocelis subcapitata, a planktonic alga and Biomphalaria glabrata, a snail) and non-target (Daphnia similis, a planktonic crustacean and Danio rerio, a fish) organisms. Test water parameters were as follows: pH = 7.4 +/- 0.1; hardness 44 +/- 1 mg/l as CaCO3; DO 8-9 mg/l at the beginning and > 4 mg/l at the end; temperature, fish and snails 25 +/- 1 degrees C, Daphnia 20 +/- 2 degrees C, algae 24 +/- 1 degrees C. D. similis (immobilization), 48-h EC50s (95% CLs) ranging from 0.013 (0.011-0.016) to 0.043 (0.033-0.057) mg Cu/l, and R. subcapitata (growth inhibition), 96-h IC50s from 0.071 (0.045-0.099) to 0.137 (0.090-0.174) mg Cu/l, were the most susceptible species. B. glabrata (lethality), 48-h LC50s from 0.179 (0.102-0.270) to 0.854 (0.553-1.457) mg Cu/l, and D. rerio (lethality), 48-h LC50s 0.063 (0.045-0.089), 0.192 (0.133-0.272) and 0.714 (0.494-1.016) mg Cu/l, were less susceptible than Daphnia to copper-based pesticides. Findings from the present study therefore suggest that increased levels of copper in water bodies is likely to adversely affect a variety of aquatic species.     

Control of common freshwater predatory fish, Channa punctatus, through Nerium indicum leaf extracts

The diethyl ether, chloroform, acetone and methanol extract of Nerium indicum leaf were evaluated for their piscicidal activity against common freshwater air breathing predatory fish Channa punctatus. The rank of order of toxicity (LC50) of the leaf extract was, diethyl ether extract (17.34 mg/l)>acetone (40.01 mg/l)>chloroform (40.61 mg/l)>and methanol (106.37 mg/l). There was a significant negative correlation between LC50 values and exposure periods. Thus increase in exposure period, LC50 decreases from 17.34 mg/l (24 h) to >13.58 mg/l (96 h) in the diethyl ether extract. Similar trends were also observed in acetone, chloroform and methanol extracts. Exposure of sub-lethal doses (40% and 80% of LC50) of the diethyl ether extract of N. indicum leaf (which has maximum piscicidal activity) for 24 or 96 h caused significant alteration in the level of total protein, total free amino acid, nucleic acid, glycogen, pyruvate, lactate and activity of enzyme protease, phosphatases, alanine aminotransferase, aspartate aminotransferase and acetylcholinesterase in liver and muscle tissue. The alterations in all the above biochemical parameters were significantly (P<0.05) time and dose dependent. There was a significant recovery in all the above biochemical parameters, in both liver and muscle tissues of fish after the seventh day of the withdrawal of treatment. Thus, the leaf extracts of N. indicum have potent piscicidal activity against fish C. punctatus and also significantly affect both aerobic and anaerobic pathway of respiration in fish.     

Acute toxicity and genotoxicity of five selected anionic and nonionic surfactants

The results of four toxicity bioassays of selected anionic and nonionic surface active agents were presented. Three widely used anionic surfactants that belong to alkyl sulphates (AS), alkylbenzene sulphonates (LAS) and alkylpolyoxyethylene sulphates (AES) as well as nonionic surfactants: polyoxyethylene alkyl ethers (AE) and polyoxylethylene alkylphenyl ethers (APE) were tested. Three different toxicity assays to aquatic organisms: Physa acuta Draparnaud, Artemia salina and Raphidocelis subcapitata were applied. Additionally, the genotoxicity test with Bacillus subtilis M45 Rec- and H17 Rec+ strains was performed. The obtained results showed that none of the surfactants studied was genotoxic at the concentration 1000 mg l(-1). On the basis of toxicity tests to aquatic organisms all tested anionic surfactants were harmful (LC50 between 10 and 100 mg l(-1)), whereas nonionic ones were toxic (LC50 between 1 and 10 mg l(-1)) or even highly toxic (LC50 below 1 mg l(-1)). Moreover, the bigger was the molecular weight of the tested compound, the higher toxicity was observed.     

The toxicity and bioavailability of atrazine and molinate to Chironomus tepperi larvae in laboratory and river water in the presence and absence of sediment

Acute (10 day) semi-static toxicity tests in which the midge, Chironomus tepperi, were exposed to atrazine and molinate were conducted in laboratory water and in river water, in the absence and presence of sediment. The bioavailability measured as median lethal concentrations (LC50) and 95% fiducial limits (FLs) of atrazine to C. tepperi in laboratory water in the absence and presence of sediment were 16.6 (14.3-19.4) and 21.0 (18.2-24.1) mg l(-1), respectively while the corresponding values in river water were 16.7 (14.7-19.0) and 22.7 (20.3-25.4) mg l(-1), respectively. For molinate, the LC50 and FL values in laboratory water in the absence and presence of sediment were 8.8 (6.8-11.4) and 14.3 (12.4-16.4) mg l(-1), respectively and the corresponding values in river water were 9.3 (7.6-11.3) and 14.5 (12.4-16.9) mg l(-1), respectively. Atrazine has low toxicity (LC50 > 10 mg l(-1)) while molinate has moderate toxicity (1 mg l(-1) < LC50 < 10 mg l(-1)) to C. tepperi. River water did not significantly (P > 0.05) reduce the bioavailability of either chemical to C. tepperi. However, the presence of sediment did significantly (P < 0.05) reduce the bioavailability of both atrazine and molinate to C. tepperi.     

In vivo evaluation of three biomarkers in the mosquitofish (Gambusia yucatana) exposed to pesticides

In this study, the acute toxicity and the in vivo effects of commercial chlorpyrifos, carbofuran and glyphosate formulations on cholinesterase (ChE), glutathione S-transferase (GST) and lactate dehydrogenase (LDH) activities of the mosquitofish (Gambusia yucatana) were investigated. In a first phase of the study, head and muscle ChE were characterized with different substrates (acetylthiocholine iodide, s-butyrylthiocholine iodide and propionylthiocholine iodide) and the selective inhibitors eserine hemisulfate, 1,5-bis(4-allyldimethylammoniumphenyl)-pentan-3-one dibromide (BW284C51), and N,N'-diisopropylphosphorodiamic acid (iso-OMPA). The results obtained suggest that the enzyme present in both head and muscle of G. yucatana is mainly acetylcholinesterase (AChE). Acute toxicity was evaluated by exposing fish to several concentrations of single pesticides and of a mixture of chlorpyrifos/glyphosate. LC50 values were determined after 96 h of exposure, except in the case of carbofuran for which LC50 was calculated after 24 h since almost all the fish died within this period. LC50 values were 0.085 mg/l for chlorpyrifos, 17.79 mg/l for glyphosate, 0.636 mg/l for carbofuran and 0.011 mg/l for the chlorpyrifos/glyphosate mixture. A Toxic Unit approach was used to compare the toxicity of chlorpyrifos and glyphosate when occurring in a mixture with their toxicities as single compounds. Synergistic effects of chlorpyrifos and glyphosate when present in a mixture were found. At the end of each bioassay (24 h for carbofuran, 96 for the other substances/mixture), effects on biomarkers were analyzed. Muscle LDH activity was not altered by any of the three pesticides tested. Gill GST activity was significantly inhibited (40%) by carbofuran after 24 h of exposure to concentrations equal or higher than 0.06 mg/l. ChE muscle and head activity were significantly inhibited (50% and 30%, respectively) by carbofuran at concentrations equal or higher than 0.25 mg/l. Chlorpyrifos induced a significant inhibition of both muscle and head ChE (80% and 50%, respectively) after 96 h of exposure to concentrations equal or higher than 0.05 mg/l. Carbofuran did not induce significant alterations of fish ChE. The ChE EC50 determined for chlorpyrifos/glyphosate mixture (0.070 mg/l) was higher than the correspondent value calculated for chlorpyrifos alone (0.011 mg/l) suggesting an antagonistic effect of glyphosate on ChE inhibition by chlorpyrifos. ChE activity of G. yucatana seems to be a good biomarker to diagnose the exposure of wild populations of this species exposed to anticholinesterase pesticides.     

Ecotoxicological Characterisation and Classification of Existing Chemicals

GOAL, SCOPE AND BACKGROUND: In 1998, the International Council of Chemical Associations (ICCA) launched a global initiative to investigate more than 1,000 HPV chemicals (High Production Volume, > or = 1,000 t/a) within the refocused OECD HPV Chemicals Programme. Up to the OECD SIDS Initial Assessment Meeting in April 2004 (SIAM 18) 147 ICCA dossiers (ca. 230 CAS-No) have been assessed based on a harmonised data set. The environmental profile and an ecotoxicological characterisation of these chemicals are presented here. Data for acute aquatic toxicity were correlated among each other, as well as data for fish (LC50, LD50) and rodents (LD50). The data for acute aquatic toxicity are compared with other existing chemicals. METHODS: Data of the ICCA HPV chemicals from the OECD SIAM 11-18 are presented for: log Kow (as an indicator for bioaccumulation potential), biodegradation, acute aquatic toxicity and availability of long-term toxicity data. Correlation analysis was performed with log transformed data and a linear regression model was fitted to the data, if a significant correlation was found. Acute toxicity for fish and acute oral toxicity for rodents were correlated on a molar basis. Acute aquatic toxicity of the chemicals is compared with data from BUA reports 1-234 and a random EINECS sample (Knacker et al. 1995). RESULTS AND DISCUSSION: According to the dossier information, 71 of the 147 ICCA chemicals are not 'readily biodegradable', 21 have a log Kow > or = 3, and 44 are 'toxic' (LC/EC50 < or = 10 mg/L) or 'very toxic' (LC/EC50 < or = 1 mg/L) to aquatic organisms. For 77, only the base set (acute fish, Daphnia and algae) is available, for the rest at least one long-term test (fish or Daphnia) is available and three tests for a mere 14 others. Based on the data presented, the SIAM gives recommendations for Environment and Human Health. 22 chemicals have been identified as a 'candidate for further work' for Environment and 16 for Human Health. The highest correlation coefficient was obtained correlating fish and Daphnia (r2 = 0.79). LC50 (fish) is significantly correlated with LD50 (rodent), but data are widely scattered. The correlation is not improved after transforming LC50 (fish) to LD50 (fish), using BCF QSAR. Based on acute aquatic toxicity, 25.1% of the chemicals from the BUA reports 1-234 are classified as 'very toxic' (LC/EC50 < or = 1 mg/L). This proportion is 2.5-fold higher than the ICCA HPV chemicals and 1.4-fold higher than the random EINECS sample. CONCLUSIONS: Correlation coefficients for aquatic toxicity data are rather uniform (0.57-0.79) compared with literature data, but also the best correlation was observed between fish and Daphnia. Because the scatter around the regression lines is still considerable, simple predictions of ecotoxicity between species are not possible. Correlation of LC50 (fish) and LD50 (rodent) indicates that toxicity is different. Surprisingly, the correlation of fish and rodent toxicity is not improved by transforming LC50 values to internal LD50s. The selection of ICCA chemicals by market significance (production volume) leads to a classification of toxicity, which is more comparable to a random sample of EINECS chemicals than to German BUA chemicals. The latter were chosen for concern (for Environment or Human Health). RECOMMENDATIONS AND OUTLOOK: Of 147 dossiers assessed between SIAM 11-18, ca. 75% were sponsored by the three following countries: Germany (42), USA (37) and Japan (33). The current output is about 50 dossiers per year (70-100 CAS-No), but a trend for an increase of output is noticeable. Industry, national authorities, and OECD work on a further development to speed up the output. The number of chemicals with 'low priority for further work' and the work recommended for the 'candidates' (mainly exposure assessment) indicate that the data presented were adequate for an initial hazard assessment according to OECD requirements. From the ICCA HPV list (n = 880, state of 1999) 44% of the chemicals have data available to cover all SIDS endpoints for Environment and only 33% for Human Health (Allanou et al. 1999). This indicates the importance of the Initiative to provide information on existing chemicals. The authors agree with the expectation "...that the scientific information provided by this global initiative will be considered as an internationally accepted and harmonised basis for further steps of chemicals management." (ICCA 2002 b).     

Acute toxicity of heavy metals towards freshwater ciliated protists

The acute toxicity of five heavy metals to four species of freshwater ciliates (Colpidium colpoda, Dexiotricha granulosa, Euplotes aediculatus, and Halteria grandinella) was examined in laboratory tests. After exposing the ciliates to soluble compound of cadmium, copper, chromium, lead, and nickel at several selected concentrations, the mortality rate was registered and the LC50 values (with 95% confidence intervals) were calculated. Large differences appeared in sensitivities of the four species to the metals. H. grandinella showed the highest sensitivity for cadmium (0.07 mg l(-1), LC50) and lead (0.12 mg l(-1), LC50), whilst E. aediculatus showed the highest sensitivity for nickel (0.03 mg l(-1), LC50). The comparison with data obtained with other species indicate that Halteria grandinella and Euplotes aediculatus are excellent and convenient bioindicator for evaluating the toxicity of waters and wastewaters polluted by heavy metals. The short time (24 h) and simplicity of the test procedure enable this test to be used in laboratory studies.     

Determination of selected fate and aquatic toxicity characteristics of acrylic acid and a series of acrylic esters

Acrylic acid, methyl acrylate, ethyl acrylate, and butyl acrylate are commercially important and widely used materials. This paper reports the results of a series of fate and aquatic toxicity studies. The mobility in soil of acrylic acid and its esters ranged from 'medium' to 'very high'. Calculated bioconcentration factors ranged from 1 to 37, suggesting a low bioconcentration potential. Acrylic acid and methyl acrylate showed limited biodegradability in the five day biochemical oxygen demand (BOD5) test, while ethyl acrylate and butyl acrylate were degraded easily (77% and 56%, respectively). Using the OECD method 301D 28-d closed bottle test, degradability for acrylic acid was 81% at 28 days, while the acrylic esters ranged from 57% to 60%. Acrylic acid degraded rapidly to carbon dioxide in soil (t1/2 < 1 day). Toxicity tests were conducted using freshwater and marine fish, invertebrates, and algae. Acrylic acid effect concentrations for fish and invertebrates ranged from 27 to 236 mg/l. Effect concentrations (LC50 or EC50) for fish and invertebrates using methyl acrylate, ethyl acrylate, and butyl acrylate ranged from 1.1 to 8.2 mg/l. The chronic MATC for acrylic acid with Daphnia magna was 27 mg/l based on length and young produced per adult reproduction day and for ethyl acrylate was 0.29 mg/l based on both the reproductive and growth endpoints. Overall these studies show that acrylic acid and the acrylic esters studied can rapidly biodegrade, have a low potential for persistence or bioaccumulation in the environment, and have low to moderate toxicity.     

Tolerance to,and avoidance of,hypoxia by the penaeid shrimp (Metapenaeus ensis)

Aquatic hypoxia caused by eutrophication may lead to mass mortality of valuable living resources such as fish and shrimp. However, there is little information on the hypoxic tolerance of penaeid shrimp, and whether they are able to avoid hypoxia. In laboratory experiments, LC50, LT50 and heart beats per minute were determined for juvenile Metapenaeus ensis at 0.5, 1.0, 2.0 and 6.0 mg O2 l(-1). The 8-h LC50, for DO was 0.77 mg O2 l(-1), while the LT50 at 0.5 mg O2 l(-1) was 399 min. Heart beat rate significantly declined when DO fell below 1.0 mg O2 l(-1). When confronted with a gradient of dissolved oxygen, M. ensis were able to avoid hypoxic areas and move to oxygenated water. M. ensis appeared to be sensitive to hypoxia, and their ability to detect and avoid hypoxia may enhance their survival in habitats where hypoxia may occur.     

Toxic effects of orimulsion on rainbow trout <Emphasis Type="Italic">oncorhynchus mykiss</Emphasis>

GOAL, SCOPE AND BACKGROUND: Orimulsion (stable emulsion of natural bitumen and water) is a new imported industrial fuel in Lithuania. No data on its toxicity to fish is freely available. The aim of this study was to investigate sensitivity of rainbow trout (Oncorhynchus mykiss) to acute and chronic toxicity of orimulsion and to estimate the Maximum Acceptable Toxicant Concentration (MATC) of orimulsion to fish. METHODS: Laboratory tests were conducted on rainbow trout in all stages of development (embryos, larvae, adults). Acute toxicity (96-hour duration) and long-term (28 or 60-day duration) tests evaluating the wide range spectrum of biological indices were performed under semi-static conditions. RESULTS AND DISCUSSION: Median lethal concentration (96-hour LC50) values and their 95% confidence intervals derived from the tests were: 0.1 (0.09-0.12) to embryos, 0.06 (0.05-0.07) to larvae and 2.22 (2.02-2.43) to adult fish, and 28-day LC50 to adult fish was found to be 0.26 (0.21-0.32) g/l of total orimulsion respectively. The acute toxicity of orimulsion to rainbow trout can be characterised by a narrow zone of toxic effect and a sharp boundary between lethal and sublethal concentrations. The lowest 'safe' or 'no-effect' concentration values of total orimulsion obtained in long-term tests were equal to 0.09 g/l to adult fish, 0.019 g/l to embryos, and 0.0017 g/l to larvae. Proposed value of 'application factor' for orimulsion was found to be equal to 0.03. Since orimulsion has the property to disperse in all water volume, its toxic effect on fish can be characterised by the combined effects of dispersion and water-soluble-fraction. CONCLUSIONS: Maximum Acceptable Toxicant Concentration (MATC) of 0.0017 g/l of total orimulsion to fish was derived from long-term tests based on the most sensitive parameter of rainbow trout larvae (relative mass increase at the end of the test). According to substance toxicity classification accepted for Lithuanian inland waters, orimulsion can be referred to substances of 'moderate' toxicity to fish. RECOMMENDATIONS AND OUTLOOK: For prediction and evaluation of toxic impact of orimulsion accident spills on fish, some recommendations should be given. Since orimulsion has the property to disperse in all water volume during short time periods, the amounts of both spilled orimulsion and polluted water should be ascertained. Once both parameters are known, the real concentration of orimulsion in the water body must be determined. Then this concentration must be compared with 'safe' concentration to fish. By use of 'application factor' 0.03, approximate MATC for other fish species can be estimated when only acute toxicity data (96-hour LC50 value) is available.     

Investigation of acute toxicity and the effect of cadmium chloride (CdCl2 . H2O) metal salt on behavior of the guppy (Poecilia reticulata)

In this study 96-h LC50 value of cadmium chloride (CdCl2 . H2O), a metal salt widely used in industry, was determined for the guppy (Poecilia reticulata, Pallas, 1859). The experiments were planned in four series of a total of 440 guppies employing the static test method of acute toxicity. 10 fish were placed in each replicate of each dose. The experiments were performed as four replicates, and behavioral changes in the guppy were determined for each cadmium chloride metal salt concentration. The data obtained were statistically evaluated by the use of EPA computer program based on Finney's Probit Analysis Method and a 96-h LC50 value for P. reticulata was found to be 30.4 mg/l in a static bioassay test system. This value was estimated to be 30.6 mg/l with Behrens-Karber's method. The two methods were in good agreement. 95% lower and upper confidence limits for the LC50 were 29.3 and 31.7 mg/l, respectively. The water temperature was kept between 21 and 23 degrees C. The behavioral changes observed in fish were, swimming in imbalanced manner, capsizing, attaching to the surface, difficulty in breathing and gathering around the ventilation filter.     

Acute and sub-chronic toxicity of four cytostatic drugs in zebrafish

The acute and sub-chronic effects of four cytostatic drugs—5-fluorouracil (5-FU), cisplatin (CisPt), etoposide (ET) and imatinib mesylate (IM)—on zebrafish (Danio rerio) were investigated. Acute tests were carried out in a static system in accordance with the OECD guideline 203 for adult fish and the draft guideline for fish embryos (FET test) in order to find the LC50 values of the four cytostatic drugs. Early-life stage toxicity test on zebrafish was conducted according the OECD guideline 210 using the cytostatic drugs 5-FU and IM in a semistatic system with the objective of investigating the sub-chronic effects of the cytostatic drugs on fish. In adult fish, the cytostatic drugs 5-FU and ET did not pass the limit test, thus, are considered non-toxic. In case of cisplatin, LC50 was calculated at 64.5 mg L^?1, whereas in case of IM, LC50 was at 70.8 mg L^?1. In the FET test, LC50 of 5-FU at 72-h post fertilization (hpf) was 2441.6 mg L^?1. In case of CisPt, LC50 was 349.9 mg L^?1 at 48 hpf and it progressively decreased to 81.3 mg L^?1 at 120 hpf. In addition, CisPt caused a significant delay in the hatch of larvae. In case of ET, LC50 values were not calculable as they were higher than 300 mg L^?1 at which concentration the substance crystallized in the solution. LC50 values of IM were 48 hpf; 158.3 mg L^?1 , 72 hpf; 141.6 mg L^?1, 96 hpf; 118.0 mg L^?1, and 120 hpf; 65.9 mg L^?1. In the Early-life Stage Test with 5-FU, embryonic deformities were not detected during the tests. Regarding mortalities, the 10 mg L^?1 concentration can be considered as LOEC, as statistically significant difference in mortalities was detected in this group alone. Concerning dry body weight and standard length, 1 mg L^?1 is the LOEC. In case of IM, the highest tested concentration (10 mg L^?1) can be considered LOEC for mortalities, however, the treatment did not have an effect on the other investigated parameters (dry and wet weight, standard length). All four cytostatic drugs were characterized by low toxicity in zebrafish in acute and sub-chronic tests.     

Lethal toxicity of Lindane on a teleost fish, Anguilla anguilla from Albufera Lake (Spain): hardness and temperature effects

This paper reports the results of toxicity tests conducted using Anguilla anguilla under three different water temperature (15, 22 and 29 degrees C) and two hardness regimes (250 and greater than 600 ppm CaCO3). The 96-h LC50 increased in the experimental medium (p less than 0.05) by an order of magnitude from 0.32 to 0.45 mg/L between 15 and 29 degrees C. However in the natural medium it is similar (p greater than 0.05) (0.54 to 0.55 mg/L) for these same temperatures. The toxicity of Lindane on eels increased when the water hardness decreased. The 24, 48, 72 and 96-h LC50 for this fish in both media is less at 15 degrees C (96-h LC50 = 0.32 and 0.55 mg/L) than at 29 degrees C (96-h LC50 = 0.45 and 0.55 mg/L). These results suggest that the toxicity of Lindane presents a negative temperature coefficient.