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.     

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 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.     

Mortality response and LC50 values for juvenile and adult crayfish, Procambarus clarkii exposed to Thiodan (insecticide), Treflan, MSMA, Oust (herbicides) and Cutrine-Plus (algicide)

Toxicities of three herbicides (Treflan, MSMA and Oust), an algicide (Cutrine-plus) and an insecticide (Thiodan) to juvenile (3.0-3.4 cm) and adult (9.0-10.0 cm) crayfish, Procambarus clarkii, were determined by 96 h static bioassays. Freshly prepared 0.01% and 1.0% aqueous stock solutions of these pesticides were diluted to desired concentrations. Mortalities were recorded each day up to 96 h. Aged tap-water was used for preparing all test solutions. Per cent mortalities were analyzed for linear regression and LC(50) values were computed by probit analysis. LC(50) values for juvenile P.clarkii in the descending order of toxicity were: 24 ppb Thiodan, 13 ppm Treflan, 101 ppm MSMA, 461 ppm Cutrine-plus, 12,174 ppm Oust; and for adults these values were: 423 ppb Thiodan, 26 ppm Treflan, 1019 ppm MSMA and 2945 ppm Cutrine-plus. No LC(50) values for adult crayfish could be computed for Oust herbicide which caused no mortalities up to 60,000 ppm concentration. Comparing the overall toxicities of these pesticides, Thiodan was the most toxic to all crayfish; followed by Treflan, MSMA, Cutrine-plus and Oust. Oust was more than 1/2 million times less toxic than Thiodan to juvenile crayfish. Published LC(50) values indicate that freshwater crayfish are more tolerant than marine decapods, Crangon septemspinosa and Mysidopsis bahia, to Thiodan insecticide and Treflan herbicide.     

Impaired glutathione redox status is associated with decreased survival in two organophosphate-poisoned marine bivalves

Biomonitoring organophosphate (OP) exposure in marine environments is generally achieved by the measurement of acetylcholinesterase activity in bivalves like mussels. However, there is evidence that indicates that oxidative stress may be implied in OP toxicity. The aim of this study was to evaluate the relationship between survival from the OP insecticide fenitrothion and glutathione levels in marine bivalves. Mussels (Mytilus galloprovincialis Lam.) and scallops (Flexopecten flexuosus Poli) were exposed, in a time to death test, to their LC85 of fenitrothion for 96 h. OP-poisoned mussels showed reduced (GSH) and oxidised (GSSG) glutathione depletion in the digestive gland, muscle and gills. Pectinid spats exposed to this insecticide presented GSH depletion in the digestive gland and mantle, and a reduction of the GSH/GSSG ratio in gills and mantle. Although survival curves were significantly different and mussels withstood twice as much fenitrothion as pectinid spats, muscular GSH/GSSG ratio was highly related to mortality in both species. We suggest that an impairment in the glutathione redox status could result in an induction of the cell death, either by apoptosis or necrosis, leading ultimately to the death of the organism. We conclude that whereas glutathione depletion can be used as a biomarker of exposure, the muscular GSH/GSSG ratio might be used as a biochemical marker of effect and individual susceptibility to mortality of marine bivalves exposed to fenitrothion or other pollutants that induce oxidative stress.     

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 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.     

Toxicity of the mosquito control insecticide phenothrin to three life stages of the grass shrimp (Palaemonetes pugio)

Phenothrin is a synthetic pyrethroid used as a contact insecticide in mosquito control programs. This study compared the toxicity of phenothrin to adult, larval and embryonic grass shrimp (Palaemonetes pugio) and examined oxidative stress responses in adult and larval grass shrimp. The adult 24-h LC50 was 0.341 μg/L (95 % confidence intervals 0.282-0.412) and the 96-h LC50 was 0.161 μg/L (95 % CI 0.128-0.203 μg/L). The larval 24-h LC50 was 0.50 μg/L (95 % CI 0.441-0.568) and the 96-h LC50 was 0.154 μg/L (95 % CI 0.139-0.170 μg/L). In the presence of sediment, the 24-h LC50 was 6.30 μg/L (95 % CI 5.00-7.44 μg/L) for adults and 0.771 μg/L (95 % CI 0.630-0.944) for larvae. The sublethal biomarkers glutathione and lipid peroxidase (LPx) were examined after 96-h phenothrin exposure at five concentrations, and there were no statistically significant differences in these levels in adults or larvae compared to controls. There was a significant downward trend in larval LPx levels. This research confirms that phenothrin is highly toxic to grass shrimp and suggests that both adult and larval grass shrimp are appropriate life stages for risk assessments.     

Investigation of acute toxicity of cadmium sulfate (CdSO4 x H2O) and behavioral changes of grass carp (Ctenopharyngodon idellus Val., 1844)

This study attempts to determine the 96-h LC50 values of CdSO4 x H2O, a metal salt widely used in industry, for grass carp (Ctenopharyngodon idellus Val., 1844). The study was carried out in two stages with 120 grass carp individuals using statistic method of aquatic toxicity. In addition, the behavioral changes of grass carp at different CdSO4 x H2O concentrations were determined. The results were evaluated using probit analysis method and the LC50 value for grass carp species was calculated as 9.42 mg/l.     

Toxicity of the mosquito control insecticide phenothrin to three life stages of the grass shrimp (Palaemonetes pugio)

Phenothrin is a synthetic pyrethroid used as a contact insecticide in mosquito control programs. This study compared the toxicity of phenothrin to adult, larval and embryonic grass shrimp (Palaemonetes pugio) and examined oxidative stress responses in adult and larval grass shrimp. The adult 24-h LC50 was 0.341 μg/L (95 % confidence intervals 0.282–0.412) and the 96-h LC50 was 0.161 μg/L (95 % CI 0.128–0.203 μg/L). The larval 24-h LC50 was 0.50 μg/L (95 % CI 0.441–0.568) and the 96-h LC50 was 0.154 μg/L (95 % CI 0.139–0.170 μg/L). In the presence of sediment, the 24-h LC50 was 6.30 μg/L (95 % CI 5.00–7.44 μg/L) for adults and 0.771 μg/L (95 % CI 0.630–0.944) for larvae. The sublethal biomarkers glutathione and lipid peroxidase (LPx) were examined after 96-h phenothrin exposure at five concentrations, and there were no statistically significant differences in these levels in adults or larvae compared to controls. There was a significant downward trend in larval LPx levels. This research confirms that phenothrin is highly toxic to grass shrimp and suggests that both adult and larval grass shrimp are appropriate life stages for risk assessments.     

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.     

Effects of the synthetic pyrethroid insecticide, permethrin, on two estuarine fish species

Limited toxicity data are available for estuarine and marine species and the widely used pyrethroid insecticide, permethrin. This study determined acute effects of permethrin on survival, lipid peroxidation, acetylcholinesterase activity, and splenocyte proliferation for two fish species found in South Carolina estuaries; juvenile red drum (Sciaenops ocellatus) and adult mummichog (Fundulus heteroclitus). Juvenile S. ocellatus were significantly more sensitive than adult F. heteroclitus to permethrin exposure, with a 96-h LC50 value of 8 μg/L determined for red drum compared to 23 μg/L for mummichog. Lipid peroxidation activity of the liver increased in permethrin-treated fish compared to control animals after 24 h and decreased after 96 h. Permethrin had no effect on acetylcholinesterase activity of the brain at the concentrations tested. Permethrin exposure significantly inhibited splenocyte proliferation, indicating an immunosuppressive effect. Most of the effects of permethrin on fish cellular stress enzymes and survival occurred at concentrations much higher than those typically measured in the environment. However, inhibition of splenocyte proliferation in juvenile red drum occurred at approximately twice that of measured permethrin concentrations in surface water. These findings may prove useful to the future management and regulation of pyrethroid insecticide use near estuarine habitats.     

Deriving freshwater quality criteria for 2,4,6-trichlorophenol for protection of aquatic life in China

Freshwater quality criteria of 2,4,6-trichlorophenol (2,4,6-TCP) were developed with particular reference to the aquatic biota in China, and based on USEPA's guidelines. Acute toxicity tests were performed on nine different domestic species indigenous to China to determine 48 h LC(50) and 96 h LC(50) values for 2,4,6-TCP. In addition, 21 d survival-reproduction test with Daphnia magna, 30 d embryo-larval test with Carassius auratus, 60 d fry-juvenile test with Ctenopharyngodon idellus, 30 d early life stage test with Bufo bufo gargarizans and 96 h growth inhibition test with Scenedesmus obliqaus were also conducted to estimate lower chronic limit and upper chronic limit values. The final acute value (FAV) was 2.01 mg/l 2,4,6-TCP. Acute-to-chronic ratios ranged from 5.01 to 12.2. The final chronic value (FCV) and the final plant value (FPV) of 2,4,6-TCP were 0.226 and 2.24 mg/l respectively. Based on FAV, FCV and FPV for 2,4,6-TCP, a criteria maximum concentration of 1.01 mg/l and a criterion continuous concentration of 0.226 mg/l were derived. The results of this study provide useful data for deriving national or local water quality criteria for 2,4,6-TCP based on aquatic biota in China.     

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.     

Effects of bensulfuron-methyl and cinosulfuron on growth of four freshwater species of phytoplankton

The acute toxicity of sulfonylurea herbicides bensulfuron-methyl and cinosulfuron was tested on the five species of freshwater phytoplankton: Scenedesmus acutus, Scenedesmus subspicatus, Chlorella vulgaris and Chlorella saccharophila. Herbicide concentrations eliciting a 50% growth reduction over 96 h (EC50) ranged from 8 to 104 mg/l for cinosulfuron and from 0.015 to 6.2 mg/l for bensulfuron-methyl. The pesticides bensulfuron-methyl, atrazine and benthiocarb were more toxic than cinosulfuron, chlorsulfuron, molinate, fenitrothion and pyridaphenthion in a toxicity study with the same algal species. The transformation of effective concentrations of bensulfuron-methyl and cinosulfuron and other pesticides, obtained from toxicity measurements, into percent of the saturation level in water is used as a first evaluation of potential hazard to aquatic systems. The herbicides cinosulfuron, methyl-bensulfuron, atrazine and chlorsulfuron were more dangerous than the herbicides benthiocarb and molinate and than the insecticides fenitrothion and pyridaphenthion, in a study of hazard evaluation. The two species of Chlorella were more tolerant to both herbicides than the two species of Scenedesmus. A potential environmental hazard of sulfonylurea herbicides to aquatic systems has to be expected even at low environmental concentrations.     

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.     

The response of Chenopodium album L. and Abutilon theophrasti Medik. to reduced doses of mesotrione

Limited toxicity data are available for estuarine and marine species and the widely used pyrethroid insecticide, permethrin. This study determined acute effects of permethrin on survival, lipid peroxidation, acetylcholinesterase activity, and splenocyte proliferation for two fish species found in South Carolina estuaries; juvenile red drum (Sciaenops ocellatus) and adult mummichog (Fundulus heteroclitus). Juvenile S. ocellatus were significantly more sensitive than adult F. heteroclitus to permethrin exposure, with a 96-h LC50 value of 8 μg/L determined for red drum compared to 23 μg/L for mummichog. Lipid peroxidation activity of the liver increased in permethrin-treated fish compared to control animals after 24 h and decreased after 96 h. Permethrin had no effect on acetylcholinesterase activity of the brain at the concentrations tested. Permethrin exposure significantly inhibited splenocyte proliferation, indicating an immunosuppressive effect. Most of the effects of permethrin on fish cellular stress enzymes and survival occurred at concentrations much higher than those typically measured in the environment. However, inhibition of splenocyte proliferation in juvenile red drum occurred at approximately twice that of measured permethrin concentrations in surface water. These findings may prove useful to the future management and regulation of pyrethroid insecticide use near estuarine habitats.     

Comparative acute toxicities of selected pesticides to Anguilla anguilla.

The acute toxicities (24, 48, 72 and 96 hr) of eight pesticides to Anguilla anguilla were determined. The organochlorine pesticide, endosulfan was the most toxic, with LC50 values in the range of 0.042 to 0.041 mg/L. Endosulfan was followed in order of decreasing toxicity by diazinon, fenitrothion, chlorpyrifos, lindane, methidathion, trichlorfon and methylparathion. When fishes were exposed to the pesticides tested they exhibited signs of restlessness, erratic swimming, convulsions and difficulty in respiration. This response was more persistent in fishes exposed to organophosphorus pesticides.     

Toxicity of the mosquito control pesticide Scourge to adult and larval grass shrimp (Palaemonetes pugio)

This study investigated the toxicity of various concentrations of technical resmethrin and Scourge on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC50 value for adult shrimp was 0.53 microg/L (95% confidence interval (CI): 0.46-0.60 microg/L), and for larval shrimp was 0.35 microg/L (95% CI: 0.28-0.42 microg/L). In the presence of sediment, technical resmethrin produced a 24-h LC50 value for adult shrimp of 5.44 microg/L (95% CI: 4.52-6.55 microg/L), and for larval shrimp of 2.15 microg/L (95% CI: 1.35-3.43 microg/L). For Scourge, the 96-h aqueous LC50 for adult shrimp was 2.08 microg/L (95% CI: 1.70-2.54 microg/L), and for larval shrimp was 0.36 microg/L (95% CI: 0.24-0.55 microg/L). The 24-h sediment test yielded an LC50 value of 16.12 microg/L (95% CI: 14.79-17.57 microg/L) for adult shrimp, and 14.16 microg/L (95% CI: 12.21-16.43 microg/L) for larvae. Adjusted LC50 values to reflect the 18% resmethrin concentration in Scourge are 0.37 microg/L (adult), 0.07 microg/L (larvae) for the 96-h aqueous test, and 2.90 microg/L (adult), 2.6 microg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge.     

Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridylphosphorothioate) is one of the organophosphate pesticides widely used in agricultural practices throughout world and irreversible inhibitor of cholinesterase in all animal species. Limited efforts have been made to study acute genotoxic effects of chlorpyrifos (CPF) in different tissues of fish using genotoxic biomarkers. Therefore, the present investigation was aimed to study the induction of DNA damage by CPF in freshwater teleost fish Channapunctatus using micronucleus assay (MN assay) and alkaline single-cell gel electrophoresis (comet assay). The value of LC(50) - 96 h of CPF was determined as 811.98 microgl(-1) for C. punctatus, in a semi-static system and on the basis of LC(50) value three acute concentrations viz., 203, 406 and 609 microgl(-1) were determined. The fishes were exposed to the different concentrations of CPF for 96 h and samplings were done at regular intervals for assessment of the MN frequencies and DNA damage. In general, significant effects (P<0.01) from both concentrations and time of exposure were observed in exposed fishes. It was found that the micronucleus induction was highest on 96 h at all concentrations in the peripheral blood. Similar trend was observed for the DNA damage measured in terms of the percentage of tail DNA in the lymphocyte and gill cells. This study explored the combined use of micronucleus assay and comet assay for in vivo laboratory studies using fresh water fish for screening the genotoxic potential of xenobiotics.