Investigation of acute toxicity of chlorpyrifos-methyl on guppy Poecilia reticulata

Static bioassays were made to determine acute toxicity of chlorpyrifos-methyl, a wide spectrum organophosphorus insecticide and potential toxic pollutant of aquatic ecosystem, Guppy fish (Poecilia reticulata). Bioassays were made at a regulated temperature of 22+/-1 degrees C and were repeated three times. Lethal doses of the insecticides were determined using LC50 software programme of U.S. EPA based on Finney's Probit Analysis statistical method. The 96 h LC50 value and 95% confidence limit of chlorpyrifos-methyl for Guppy was estimated as 1.79 (1.47-2.10) mg/l. The fish exposed to chlorpyrifos-methyl exhibited behavioral changes in the form of neurotoxin toxicity: less general activity than control group, loss of equilibrium, erratic swimming and staying motionless at a certain location generally at mid-water level for prolonged periods. The 1 mg/l (lowest) concentration had similar behavior (NOEC) with the control group.     

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.     

Acute toxicity of alpha-cypermethrin to guppy (Poecilia reticulata, Pallas, 1859)

Alpha-cypermethrin is a synthetic pyrethroid insecticide used to control pests in domestic, industrial and agricultural situations. Adult male guppy fish (Poecilia reticulata standard test species) were selected for the bioassay experiments. The experiments were performed in four series and the 96-h LC50 value was determined for guppies. The acute toxicity experiments were carried out by static method and behavioral changes in guppies were determined for each alpha-cypermethrin (98% technical grade) concentration. Data obtained from the alpha-cypermethrin investigation were evaluated by the use of probit analysis statistical method and the 96-h LC50 value for guppy was estimated as 9.43 microg/l.     

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.     

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.     

Acute toxicity of the insect larvicide abate® (temephos) on the fish tilapia melanopleura and the dragonfly larvae neurocordelia virginiensis.

Abstract

Acute bioassay tests on the toxic effects of the insect larvicide Abate® (temephos) on the mouth brooder cichlid fish Tilapia melanopleum and the dragonfly larvae (Odonata) Neurocordulia virginiensis were conducted in static non renewal toxicity test set ups. The 96h‐LC50 (95% confidence intervals) was 30.2 (20.5‐ 44.20) mg/L for the fish and 2.0 (1.16–2.0) mg/L for the dragonfly larvae. The dragonfly larvae were 15 times more susceptible to the larvicide than the tilapia. The calculated NOEC (No Observable Effect Concentration) was 14.1 mg/L for the fishand less than 1.0 mg/L for the insect larvae. The estimated ‘safe’ concentration of the pesticide to the fish was 3.0 mg/L and 0.2 mg/L for insect larvae. These figures are far above the concentrations approved for use in the control of mosquito larvae(0.0004–0.01 mg/L). It appears that the application of toxic levels of the insecticide for the elimination of some aquatic invertebrates may be ‘safe’ for normal survival, growth and reproduction offish and some aquatic insect larvae.     

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.     

Comparative acute toxicity of two phytosanitary molecules,lambda-cyhalothrin and acetamiprid,on Nile Tilapia (Oreochromis Niloticus) juveniles

Abstract

This study aimed to compare the toxicity for fish of two active ingredients (lambda-cyhalothrin-20?g L^?1, a pyrethroid, and acetamiprid-15?g L^?1, a neonicotinoid) which are components of a commercial insecticide (Acer 35 EC) used in cotton crop in many West African countries. The juveniles of Oreochromis niloticus (4.01?±?0.34?g, mean body weight) were exposed for 96?h to increasing concentrations of active ingredients (lambda-cyhalothrin and acetamiprid) or a mixture similar to Acer 35 EC (composed by 20?g of chemical compound lambda-cyhalothrin and 15?g of acetamiprid dissolved in 1?L of acetone). The experiments were carried out under controlled conditions in aquaria according to OECD Guidelines. During the experiments, the behavioral responses (loss of balance, color change, hyperactivity, etc.) that usually precede death were observed in exposed fish. Mortalities were recorded in each aquarium and the LC₅₀-96h of each chemical was determined. The LC₅₀-96h obtained were respectively 0.1268, 0.0029, 182.9 and 0.5685?ppm for Acer 35 EC, lambda-cyhalothrin, acetamiprid and mixture. All insecticides used in this study had profound impact on Nile tilapia behavior which may confirm the neurotoxicity of each single active compound as well as of their mixture.     

Investigation of acute toxicity of cadmium chloride (CdCl2 . H2O) metal salt and behavioral changes it causes on water frog (Rana ridibunda Pallas, 1771)

The 96-h LC(50) value of cadmium chloride (CdCl(2).H(2)O), a metal salt widely used in industry, was determined in the water frog (Rana ridibunda Pallas, 1771). The experiments were conducted in two series and a total of 140 frogs were used to determine acute toxicity. In addition, behavioral changes in the water frog were determined for each cadmium chloride concentration. Data obtained from the cadmium chloride investigation were evaluated by using the probit analysis statistical method and the 96 h LC(50) value for water frog was estimated to be 51.2 mg/l.     

Acute and Chronic Toxicity of Chlorine Dioxide (ClO2) and Chlorite (ClO2ˉ) to Rainbow Trout (Oncorhynchus mykiss) (4 pp)

Goal, Scope and Background Chlorite (ClO2ˉ) is a primary decomposition product when chlorine dioxide (ClO2) is added during water treatment; therefore the toxic effects of both compounds on aquatic organisms are possible. Limited data are available concerning their toxicity to fish. The aim of this study was to investigate sensitivity of rainbow trout to acute and chronic toxicity of chlorine dioxide and chlorite, and to estimate the Maximum-Acceptable-Toxicant-Concentration (MATC) of those compounds in fish. Methods The acute and chronic toxicity of chlorine dioxide and chlorite to larval and adult rainbow trout was investigated in 96-hour to 20-day laboratory exposures evaluating the wide range spectrum of biological indices under semi-static conditions. Results and Discussion Median lethal concentration (96-hour LC50) values derived from the tests were: 2.2 mg/l for larvae; 8.3 mg/l for adult fish and 20-day LC50 for larvae was 1.6 mg/l of chlorine dioxide, respectively. Chlorite was found to be from 48 to 18 times less acutely toxic to larvae and adult fish, correspondingly. Both chemical compounds induced similar toxic effects in rainbow trout larvae during chronic tests (they affected cardio-respiratory and growth parameters), but chlorine dioxide had a higher toxic potency than chlorite. A significant decrease in the heart rate and respiration frequency of larvae was established. However, within an increase in exposure duration recovery of cardio-respiratory responses was seen to have occurred in larvae exposed to chlorite. Meanwhile, in larvae exposed to chlorine dioxide, a significant decrease in cardio-respiratory responses remained during all 20-day chronic bioassays. Chlorine dioxide also more strongly affected growth parameters of rainbow trout larvae at much lower test concentrations. Decreased rate of yolk-sack resorption occurred only in the tests with chlorine dioxide. Conclusions Maximum-Acceptable-Toxicant-Concentration (MATC) of 0.21 mg/l for chlorine dioxide and of 3.3 mg/l for chlorite to fish was derived from chronic tests based on the most sensitive parameter of rainbow trout larvae (growth rate). According to substance toxicity classification accepted for Lithuanian inland waters, chlorine dioxide and chlorite can be referred to substances of \moderate\ toxicity to fish. Recommendations and Outlook Due to its very reactive nature, chlorine dioxide is rapidly (in a few hours) reduced to chlorite, which is persistent also as a biocide but 16 times less toxic to fish, according to MATC. Therefore, it is much more likely that fish will be exposed to chlorite than to chlorine dioxide in natural waters. Presently accepted, the Maximum-Permitted-Concentration of total residual chlorine (TRC) in waste-water discharging into receiving waters is 0.6 mg/l. If this requirement will not be exceeded, it is unlikely that fish would be exposed to lethal or even to sublethal concentrations of chlorine dioxide or chlorite. Furthermore, chlorine dioxide does not generate toxic nitrogenous (chloramines) or carcinogenic organic residuals (trihalomethanes). All these properties make chlorine dioxide a more promising biocide than chlorine.     

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.     

Comparative study on the environmental risk induced by several pyrethroids in estuarine and freshwater invertebrate organisms

The acute toxicity of permethrin, resmethrin, and cypermethrin to four species of aquatic non-target invertebrate organisms, found in estuarine and freshwater ecosystems, was evaluated. Artemia franciscana and Brachionus plicatilis larvae, as estuarine organisms, and Brachionus calyciflorus and Thamnocephalus platyurus larvae, as freshwater organisms, were exposed for 24 h to concentrations of these pyrethroids, and the LC(50) values were compared. The freshwater organisms were more sensitive to these pyrethroids than estuarine organisms tested. A. franciscana larvae were more tolerant organisms than B. plicatilis larvae. The freshwater organisms tested have demonstrated to be a good alternative to the standard acute toxicity assays using Daphnia, although Brachionus plycatilis larvae were more sensitive to these pyrethroid insecticides than T. platyurus. Analysis of 24 h LC(50) values of these pyrethroids, determined by static bioassays, revealed that the rank order of toxicity was: permethrin相似文献   

Studies on lethal concentrations and toxicity stress of some xenobiotics on aquatic organisms

Three widely used xenobiotics pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (Butachlor) are evaluated for acute toxicity and stress behavior on freshwater fish (Heteropneustes fossilis, Clarias batrachus, Channa punctatus) and mosquito larvae (Culex pipiens fatigans). The experiment was carried out by medium treatment using intermittent flow-through system. Median lethal concentrations (LC50) were calculated by probit analysis. The LC50 values and 95% confidence intervals showed variable range for tested chemicals. Mosquito larvae generally appeared resistant than fish, while H. fossilis was found to be most sensitive. Stress signs in the form of behavioral changes are also observed. Both types of organisms are recommended as good bioindicator for the risk assessment of aquatic environment due to chemicals tested.     

Impact of cypermethrin on stream fish populations under field-use in biotech-soybean production

The impact of biotech-soybean technology on freshwater ecosystems is being evaluated in the Rolling Pampas region, Argentina. The effect of cypermethrin, the main soybean insecticide, on low-order temperate-stream fish populations was investigated for two consecutive crop cycles under field-use conditions in biotech-soybean production. Cypermethrin was unable to induce mortality or behavioral effects on any of the fish species resident in a first-order stream across a crop field (pulsed acute exposure scenario) sprayed according to conventional practices. No spatially or temporally dependent effects were observed on population parameters (size-class structure, abundance, survival, sex and immature/mature ratio, condition factor) of resident or caged Cnesterodon decemmaculatus after spraying or rainfall events, not even one year after, at the beginning of the next crop cycle. Although cypermethrin was "very highly toxic" to C. decemmaculatus in laboratory water (96h-LC(50)=0.43microg/l), its toxicity was reduced in filtered (78%) and unfiltered (92%) stream water. Changes in LC(50) values were mainly correlated with the OC content of each water fraction (r(2)=0.99; p<0.01; n=9), showing that both DOC and TOC contributed proportionally to toxicity reduction. Protective effects of stream water (12-fold reduction LC(50) values) explained the lack of effects on fish populations in the field, despite cypermethrin water concentrations after spraying reached values comparable with the 96h-LC(50). Therefore, cypermethrin under field-use conditions in transgenic-soybean production represents a low risk of acute exposure for fish populations inhabiting low-order temperate-streams rich in TOC. The relationship between LC(50) and TOC could be a convenient way to improve risk estimation based on laboratory toxicity testing.     

Acute toxicity of chlorpyrifos to embryo and larvae of banded gourami Trichogaster fasciata

This study elucidated the acute toxicity of chlorpyrifos on the early life stages of banded gourami (Trichogaster fasciata). To determine the acute effects of chlorpyrifos on their survival and development, we exposedthe embryos and two-day-old larvae to six concentrations (0, 0.01, 0.10, 1.0, 10 and 100 µg L^?1) of chlorpyrifos in plastic bowls. Log-logistic regression was used to calculate LC10 and LC50 values. Results showed that embryo mortality significantly increased with increasing chlorpyrifos concentrations. The 24-h LC10 and LC50 values (with 95% confidence limits) of chlorpyrifos for embryos were 0.89 (0.50–1.58) and 11.8 (9.12–15.4) µg L^?1, respectively. Hatching success decreased and mortality of larvae significantly increased with increasing concentrations of chlorpyrifos. The 24-h LC10 and LC50 values (with 95% confidence limits) of chlorpyrifos for larvae were 0.53 (0.27–1.06) and 21.7 (15.9–29.4) µg L^?1, respectively; the 48-h LC10 and LC50 for larvae were 0.04 (0.02–0.09) and 5.47 (3.77–7.94) µg L^?1, respectively. The results of this study suggest that 1 µg L^?1 of chlorpyrifos in the aquatic environment may adversely affect the development and the reproduction of banded gourami. Our study also suggests that banded gourami fish can serve as an ideal model species for evaluating developmental toxicity of environmental contaminants.     

Biochemical effects of clomazone herbicide on piava (Leporinus obtusidens)

This study aims to verify the effects of the clomazone concentration used in rice fields on acetylcholinesterase (AChE), thiobarbituric acid reactive substances (TBARS), protein carbonyl and catalase activity in tissues of piava (Leporinus obtusidens). LC(50)-96h was 5.0 mg L(-1) and the fish were exposed to 1/10 of LC(50)-96 h: 0.5 mg L(-1) of clomazone for 96 and 192h. The same parameters were also assayed after a recovery period of 192 h in clean water. AChE activity was reduced only in the brain and heart of fish exposed for 96 h. AChE activity was decreased in the brain, muscle and heart tissues after 192 h of exposure. After 192 h of recovery period, AChE activity remained diminished in brain and muscle and showed a decrease in eye. However, after 192 h of recovery, AChE activity in heart was recovered. Fish showed increased TBARS levels in brain at all experimental periods. TBARS levels decreased in liver and muscle tissues after 192 h of exposure. The increase in muscle TBARS persisted in fish transferred to clean water. Protein carbonyl in the liver was increased in all periods studied including the recovery period. Catalase activity was reduced during all periods. The present study demonstrates the occurrence of disorders in AChE, TBARS, protein carbonyl and catalase activity in piava. The results also show changes in fish after exposure to an environmentally relevant concentration of clomazone. Most effects observed persisted after the recovery period. Thus, these parameters may be used to monitor clomazone toxicity in fish.     

Comparison of in vitro and in vivo acute fish toxicity in relation to toxicant mode of action

Acute toxicity to fish hepatoma cell line PLHC-1 and to juvenile rainbow trout was examined for 18 plant protection products. The main objective was to explore whether hepatoma cells could be used to predict acute toxicity in fish taking into account the mode of toxic action and compound properties. Acute fish toxicity was determined using the OECD guideline test 203 and compared to predicted baseline LC50 of acute fish toxicity calculated with a quantitative structure-activity relationship (QSAR) derived for guppy fish. Cytotoxicity was determined through the inhibition of neutral red uptake (NR(50)) into lysosomes and compared to predicted baseline cytotoxicity derived for goldfish GFS cells. In general, NR50 values were higher by a factor ranging from 3 to 3000 than the corresponding acute LC50. A weak correlation between NR50 and LC50 values was found (log/log: r2=0.62). Also the lipophilicity (log K(ow)) was not a good predictor for cytotoxicity (r2=0.43) and lethality (r2=0.57) of these pesticides. The neutral red assay is detecting general baseline toxicity only. Comparing LC50 data to QSAR results, the compounds can be classified to act as narcotics or reactive compounds with a specific mode of toxic action in fish. The results indicate that limitation of the neutral red assay in predicting acute fish toxicity. A promising alternative might be the assessment of toxicity in a set of in vitro systems addressing also cell-specific functions which are related to the mode of toxic action of the compound.     

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.     

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.