Effect of fenvalerate on oxidative stress biomarkers in the brackish water prawn Penaeus monodon

The toxicity of fenvalerate to the prawn Penaeus monodon was evaluated using biomarkers of stress. In a preliminary bioassay test, P. monodon was exposed to a series of fenvalerate concentrations, which showed 4, 6.5 and 8.5 μg L⁻¹ to be sublethal, median lethal and lethal, respectively. Sublethal effect of fenvalerate was further evaluated in hepatopancreas, muscle and gills of prawns with reference to oxidative stress biomarkers. Significant induction of lipid peroxidation and glutathione-S-transferase activity was found in hepatopancreas, muscle and gills of prawns exposed to fenvalerate when compared to control (P < 0.001, P < 0.05 and P < 0.05). On the contrary, the activities of Superoxide dismutase, catalase, glutathione peroxidase, vitamin C, vitamin E and glutathione were found to be reduced in the experimental group of prawns when compared to control. The results suggest that the animals were under oxidative stress when exposed to sublethal concentration of fenvalerate.     

Induction of heat shock protein genes by chlorfenapyr in cultured cells of the cabbage armyworm, Mamestra brassicae

Effects of structurally different insecticides, permethrin, chlorfluazuron, chlorfenapyr, prothiofos, methomyl, and thiocyclam on expression of hsp90, hsp70, hsp20.7, and hsp19.7 were examined using cultured cells of the cabbage armyworm, Mamestra brassicae. Significant induction of hsp90, hsp70, hsp20.7, and hsp19.7 expression was observed in response to chlorfenapyr. No induction was observed for the remaining five insecticides. The chlorfenapyr-induced expression was time-dependent and concentration-dependent. A significant reduction in expression levels of hsp70, hsp20.7, and hsp19.7 was observed in the time course when the cells exposed to chlorfenapyr were transferred to chlorfenapyr-free medium. These results suggest that hsp70, hsp20.7, and hsp19.7 might be useful to assess cellular distress or injury by chlorfenapyr.     

Carbaryl induced alterations in the reproduction and metabolism of freshwater snail Lymnaea acuminata

When the freshwater snail Lymnaea acuminata was exposed to sub-lethal doses (2.0, 5.0, and 8.0 mg/L) of carbaryl, fecundity was significantly reduced and even stopped at higher sub-lethal doses and altered metabolic activity in the body tissue of the snail was observed. The change from aerobic to anaerobic metabolism results in lesser energy production in the body tissues of the snails, causing paralysis and finally death. This reduced fecundity and altered metabolism suggests that it would be better to avoid the use of carbaryl pesticides in the water bodies or fields adjoining the water bodies particularly in the rainy season.     

Acute toxicity of organochlorine insecticide endosulfan and its effect on behaviour and some hematological parameters of Asian swamp eel (Monopterus albus, Zuiew)

In Asia, Monopterus albus (Asian swamp eel) is commonly found in rice fields, muddy ponds and swamp areas. Because of its habitat, it is exposed to toxic pesticides used in rice fields, especially endosulfan, which is one of the most widely used insecticides. This study aims to determine the acute toxicity of endosulfan and its effect on the behaviour and some hematological parameters of the fish. The 96 h-LC₅₀ (with 95% confidence limits) obtained in this study for M. albus was 0.42 (0.35-0.50) μg L⁻¹. After 96 h exposure to endosulfan, the fish exhibited a series of abnormal behavioural responses; these included imbalanced position, restlessness of movement, erratic swimming, tremor, flashing and lethargy. When the swamp eels were exhausted, they were laterally recumbent on the bottom and had no opercula movement. Endosulfan toxicity also caused a significant lower (p < 0.05) value of erythrocyte count, leukocyte count, hemoglobin and hematocrit as compared to the control experiment. Nevertheless, the erythrocyte cell showed an increasing trend in size. The swollen erythrocyte cells may be impaired in their oxygen-carrying capacity. Severe blood loss through the gill capillary and hematemesis of the fish may be the main reasons for the hematological changes in the test organisms. Despite its relatively bigger size (30-40 cm in length) as compared to most of the other test organisms (<5 cm in length) for endosulfan toxicity, it is interesting to find that this fish is one of the most sensitive freshwater fishes to endosulfan.     

Effects of carbaryl and azinphos methyl on juvenile rainbow trout (Oncorhynchus mykiss) detoxifying enzymes

In this study, the effects of sublethal exposures to the anticholinesterase insecticides azinphos methyl (AzMe) and carbaryl on the detoxifying responses of juvenile rainbow trout Oncorhynchus mykiss were investigated. Juvenile specimen were exposed to sublethal concentrations of AzMe (2.5 and 5 μg/L) and carbaryl (1 and 3 mg/L) for 24, 48 and 96 h. Carboxylesterase (CbE), catalase (CAT) and glutathione S-transferase (GST) activities as well as reduced glutathione (GSH) and cytochrome P450-1A (CYP1A) levels were monitored in liver and/or kidney. In all exposed groups liver CbE was significantly inhibited. Liver and kidney GSH level was reduced after sublethal exposure to both compounds. Carbaryl induced CAT activity during the first 48 h of exposure, followed by a significant decrease, whereas AzMe continuously decreased CAT activity. GST activity and CYP1A were transiently induced at 24 h by carbaryl exposure (3 mg/L) but sublethal exposure to AzMe did not affect GST activity or CYP1A. Our results show that the O. mykiss detoxifying system are a target for carbaryl and AzMe action, probably affecting redox balance. Although the responses showed similar trends in both organs, they were more important in liver than in kidney. The early inhibitory effect in CAT activity and GSH content produced by AzMe may be associated with a high degree of oxidative stress. Early induction of CYP1A, GST and CAT by carbaryl followed by enzyme inhibition suggests a milder or delayed oxidative stress, revealing differences between both pesticides metabolization. CbE inhibition is a good biomarker for AzMe and carbaryl exposure.     

Induction of heat shock protein genes by chlorfenapyr in cultured cells of the cabbage armyworm,Mamestra brassicae

Effects of structurally different insecticides, permethrin, chlorfluazuron, chlorfenapyr, prothiofos, methomyl, and thiocyclam on expression of hsp90, hsp70, hsp20.7, and hsp19.7 were examined using cultured cells of the cabbage armyworm, Mamestra brassicae. Significant induction of hsp90, hsp70, hsp20.7, and hsp19.7 expression was observed in response to chlorfenapyr. No induction was observed for the remaining five insecticides. The chlorfenapyr-induced expression was time-dependent and concentration-dependent. A significant reduction in expression levels of hsp70, hsp20.7, and hsp19.7 was observed in the time course when the cells exposed to chlorfenapyr were transferred to chlorfenapyr-free medium. These results suggest that hsp70, hsp20.7, and hsp19.7 might be useful to assess cellular distress or injury by chlorfenapyr.     

Biochemical and histological hepatic changes of Nile tilapia Oreochromis niloticus exposed to carbaryl

The purpose of this study was to evaluate biochemical and morphological responses induced by carbaryl in the liver of Nile tilapia (Oreochromis niloticus) exposed during 21 days to sublethal concentrations (0.25 and 0.5 mg L⁻¹), testing also recover for 14 days in clean water, after 14 days exposure. The activities of the following enzymes were measured: superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR), and reduced (GSH) and oxidized glutathione (GSSG). Globally, our data showed that exposure to carbaryl decreased the SOD, CAT, GR, and GST activities, except for the SOD and GST activities after 14 days exposure to 0.25 mg L⁻¹. In contrast, after 14 days exposure the GR activity of the hepatic tissue from carbaryl-treated fish showed significant elevation in relation to the control. When fish were left to recover, a positive response was seen in the GSH and GSSG contents. The results of the recovery group suggest that the toxicity produced by carbaryl is reversible to some extent within 15 days. The liver histological analysis showed differences between fish concerning the cellular vacuolization degree (VD) of the hepatocytes. In fish exposed to carbaryl it was observed an increasing hepatocellular basophilia. No other histological alterations were observed when fish was exposed to carbaryl, except a few necrotic foci at day 7. The sections stained with PAS reaction showed that the vacuolization was always not due to glycogen deposits, thus suggesting lipid accumulation. The combined increased basophilia and glycogen depletion is a common, although non-specific, liver response to many toxicants. In short, this work shows a relation between histological and biochemical changes in liver and carbaryl exposure. The effects of carbaryl were observed at different concentrations.     

Biochemical alteration induced by monocrotophos in the blood plasma of fish, Channa punctatus (Bloch)

Monocrotophos (MCP), commonly known as azodrin, is one of the organophosphate (OP) pesticides extensively used in agricultural practices throughout the world. Channa punctatus were exposed to sublethal concentrations (0.96 and 1.86 mg/L) of monocrotophos for 15 and 60 days to assess the alterations in the level of some biochemical parameters in blood plasma. Significant alterations in all the biochemical parameters were found to be dose dependent. Hypoglycemia and hypocholesteremia were observed in plasma of fish at both exposure periods (15 and 60 days). Increased activities of glutamate-oxalacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), acid and alkaline phosphatase of blood plasma indicated hepatic tissue damage. Decrease in lactate dehydrogenase (LDH) content in plasma further indicated lower metabolic rate after 60 days of exposure. Significant decline in triglycerides content was observed in fish exposed to both sublethal concentrations of monocrotophos. We suggest that analysis of biochemical parameters in the fish blood may be useful in environmental biomonitoring.     

Oxygen consumption and ammonia excretion of the freshwater crab Trichodactylus borellianus exposed to chlorpyrifos and endosulfan insecticides

The effects of lethal and sublethal concentrations of chlorpyrifos and endosulfan on oxygen consumption and ammonia excretion rate of the crab Trichodactylus borellianus were evaluated. Oxygen consumption and energy expenditure had significant effect in relation to exposure times. Regarding endosulfan, a significant difference in consumption among times of exposure was registered in 625 μg L⁻¹. Moreover, at the highest concentration, energy expenditure rate was observed stabilized during 1-3 h. A significant increase in ammonia excretion was evidenced in 150 and 300 μg L⁻¹ of chlorpyrifos. The O:N ratio showed a decrease in chlorpyrifos and in 2500 μg L⁻¹ of endosulfan. This indicated a shift towards protein primary metabolism. An increment in the O:N ratio was observed in the lower endosulfan solutions. The relation oxygen:nitrogen showed a shift towards lipid and carbohydrate primary metabolism. This work indicated the complexity of the metabolism in the freshwater crab affected by xenobiotic elements.     

Toxic effects of novel organophosphorus insecticide (RPR-V) on certain biochemical parameters of euryhaline fish, Oreochromis mossambicus

The euryhaline fish, Oreochromis mossambicus was exposed to sub-lethal concentration (0.017 mg L⁻¹) of a novel phosphorothionate, 2-butenoic acid-3-(diethoxy phosphinothionyl) ethyl ester (RPR-V) for 30 days and allowed to recover for 7 days. Important biomarker enzymes were assayed in plasma, brain, gill, liver, kidney, and muscle during exposure tenures of day-3, -7, -15, -30, and also at 7 days (withdrawal) after stopping treatment. Acetylcholinesterase (AChE) activities of brain, gill, and muscle were strongly inhibited by 67, 75, and 66%, respectively, on day-30. Exposure (time) dependent increases in alanine aminotransferase (ALAT), and aspartate aminotransferase (ASAT), acid phosphatase (AcP), and alkaline phosphatase (AkP), activities in plasma and kidney; AcP and AkP activities in gill were noticed. However, significant decrease in ALAT, ASAT, AcP, and AkP activities in liver was observed. The depletion of glycogen was observed in liver, brain, and gill tissues, an indication of typical stress related response of the fish with pesticide. A significant increase in lactate dehydrogenase (LDH) activity in gill and brain was observed and decreased in liver and muscle, indicating tissue damage and muscular harm. Depletion of glutathione (GSH) was observed in the above tissues, there by enhancing the lipid peroxidation resulting in cell damage. The induction in hepatic glutathione-S-transferase (GST) levels indicates the protection against the toxicity of xenobiotic-induced lipid peroxidation. There was a significant recovery in all the above biochemical parameters, in all the tissues of fish after a recovery period of 7 days. These results revealed that RPR-V affects the intermediary metabolism of O. mossambicus and the increase of biomarker enzymes in plasma, might be due to the necrosis of liver.     

Behavioral, morphological deformities and biomarkers of oxidative damage as indicators of sublethal cypermethrin intoxication on the tadpoles of D. melanostictus (Schneider, 1799)

Concerns have been raised that the amphibian larval stages are particularly at risk and may be vulnerable to adverse effects of pesticides. The present study reports acute toxicity of cypermethrin at 24, 48, 72 and 96 h through static renewal bioassay test for Duttaphrynus melanostictus. The LC₅₀ values were 5.15, 4.55, 3.95, and 3.34 μg/L for 24, 48, 72, and 96 h respectively. At sublethal concentration (0.33 μg/L) behavioral, morphological and biochemical changes were studied. The behavioral and morphological anomalies observed in the present study are typical signs of cyano pyrethroid poisoning. Significant changes were observed in total, soluble, and structural proteins. The depletion of all the protein fractions observed in this investigation led to progressive protein oxidation and catabolism of proteins. Decreased protein level has resulted in a marked elevation of free amino acid levels at all time intervals. The induction of catalase, glutathione-S-transferase activities and elevation in the levels of hydrogen peroxide, reduced glutathione, and malondialdehyde eventually lead to oxidative damage of biomolecules, showing that the generation of reactive oxygen species and oxidative stress are involved in the toxicity induced by cypermethrin. Indicating increased susceptibility of tadpoles. Thus, an exposure to cypermethrin at sublethal concentration had catastrophic effect on tadpoles of D. melanostictus.     

Cell death localization in situ in laboratory reared honey bee (Apis mellifera L.) larvae treated with pesticides

In this study, cell death detected by DNA fragmentation labeling and phosphatidylserine (PS) localization was investigated in the honey bee (Apis mellifera L.) midgut, salivary glands and ovaries after treating larvae with different pesticides offered via an artificial diet. To do this, honey bee larvae reared in an incubator were exposed to one of nine pesticides: chlorpyrifos, imidacloprid, amitraz, fluvalinate, coumaphos, myclobutanil, chlorothalonil, glyphosate and simazine. Following this, larvae were fixed and prepared for immunohistologically detected cellular death using two TUNEL techniques for DNA fragmentation labeling and Annexin V to detect the localization of exposed PS specific in situ binding to apoptotic cells. Untreated larvae experienced ∼10% midgut apoptotic cell death under controlled conditions. All applied pesticides triggered an increase in apoptosis in treated compared to untreated larvae. The level of cell death in the midgut of simazine-treated larvae was highest at 77% mortality and statistically similar to the level of cell death for chlorpyrifos (65%), imidacloprid (61%), myclobutanil (69%), and glyphosate (69%) treated larvae. Larvae exposed to fluvalinate had the lowest midgut columnar apoptotic cell death (30%) of any pesticide-treated larvae. Indications of elevated apoptotic cell death in salivary glands and ovaries after pesticide application were detected. Annexin V localization, indicative of apoptotic cell deletion, had an extensive distribution in the midgut, salivary glands and ovaries of pesticide-treated larvae. The data suggest that the tested pesticides induced apoptosis in tissues of honey bee larvae at the tested concentrations. Cell death localization as a tool for a monitoring the subclinical and sub-lethal effects of external influences on honey bee larval tissues is discussed.     

Metabolic changes in the air-breathing fish Anabas scandens on long-term exposure to endosulfan

The long-term effect of endosulfan on the carbohydrate metabolites of the fish Anabas scandens was studied. Exposure of fish to a sublethal concentration of 6 ppb, for a period of 21 days resulted in marked changes in total carbohydrate, glycogen, lactic acid, and pyruvic acid content of the tissues. Lactate levels were found to be elevated, whereas pyruvate and glycogen content of the tissues declined. The results indicate that exposure of fish to endosulfan leads to metabolic adaptations to suit the functional need of the animal.     

Changes in metabolic profiles of urine from rats following chronic exposure to anticholinesterase pesticides

Previous studies have demonstrated that the anticholinesterase pesticides chlorpyrifos and carbaryl are neurotoxic to mammals. However, the toxicity of these pesticides to other organs and their potential interactive effects remain unclear. Our goal in this study was to assess the toxicities of ingestion of chlorpyrifos and carbaryl both separately, and in combination to non-nervous systems, especially the effect on urinary metabolic profiles, in rats. Chlorpyrifos, carbaryl and a mixture of these pesticides, were administered orally to Wistar rats for 90 consecutive days. Histopathological examination of liver and kidney and metabonomic analysis based on the urinary ¹H nuclear magnetic resonance spectra were used to investigate the toxic effects. The results showed that no histopathological changes were observed in the liver or kidney tissues, but metabonomic analysis revealed alternations in a number of urinary metabolites involving in the energy metabolism in liver mitochondria. Treatment of rats with chlorpyrifos alone led to an increase in creatine, glycine, dimethylglycine, dimethylamine, glutamine, succinate, alanine, lactate, and glucose. The categories of main differential urinary metabolites in carbaryl-treated rats were similar to those in chlorpyrifos-treated rats, whereas the changes were of varying degree. A combination of a low dose of chlorpyrifos and carbaryl resulted in an increase in the levels of main urinary metabolites compared to the controls, and the increase in signal intensity of the main metabolites was lower than that in the rats exposed to chlorpyrifos or carbaryl alone. All above results suggest that chronic exposure to chlorpyrifos and carbaryl alone, or in combination could cause disturbance of metabolic function in liver mitochondria and renal failure. Overall, we have shown that urine metabonomic analysis is non-invasive, sensitive, and relatively fast for assessing the individual or mutual effects following exposure to pesticides.     

Toxicity and metabolism of endosulfan and its effect on oxygen consumption and total nitrogen excretion of the fish Macrognathus aculeatum

Experiments were conducted with the freshwater fish Macrognathus aculeatum to study the toxicity and metabolism of endosulfan and the effect of the pesticide on the oxygen consumption and total nitrogen excretion. The 96-hr LC₅₀ value was 3.5 ± 0.2 ppb. In brain, gills, gut, liver, and kidney, endosulfan was metabolized to endosulfan sulfate, but this appears to be only an intermediary step as the nontoxic endosulfan ether was found only in the liver and kidney, the principal organs of elimination of toxicants in fish. The pesticide, both at sublethal and lethal concentrations, decreased oxygen consumption and total nitrogen excretion.     

Effect of pesticides on spiders occurring on apple and citrus in Israel

Field experiments in an apple orchard and in a citrus grove were carried out to evaluate the effect of four commercial pesticides in common use in Israel against apple and citrus pests, on the spider populations inhabiting the trees. The spider populations on apple were markedly suppressed by the pesticides, the order of toxicity being Talstar (biphenate) >Mavrik (fluvalinate) > Smash (fenpropathrin) > Dursban (chlorpyrifos). When grapefruit trees were treated with carbaryl + formothion, 232 spiders were sampled in the unsprayed plot, 55 days after treatment, as compared with only 11 spiders in the treated plot. Two and 7 days after treatment with chlorobenzilate, the sample from the treated plot numbered 68 and 55 spiders, respectively, as compared with 50 spiders collected 24 h before treatment. In addition, laboratory tests were carried out to determine the susceptibility of the spiderChiracanthium mildei L. Koch to 17 pesticides. When the spiders were exposed to grapefruit leaves which had been dipped 1 h previously for 5 sec in the aqueous emulsions of the pesticides, chlorpyrifos, fenpropathrin, fenvalerate, phosphamidon and biphenate caused 100%, and cypermethrin and fluvalinate 60% mortality, whereas all the other pesticides tested - acaricides, fungicides and herbicides - caused about 10-40% mortality.     

In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills

Here we investigated the in vitro and in vivo effects of the pesticides, deltamethrin, diazinon, propoxur and cypermethrin, on the activity of rainbow trout (rt) gill carbonic anhydrase (CA). The enzyme was purified from rainbow trout gills using Sepharose 4B-aniline-sulfanilamide affinity chromatography method. The overall purification was approx. 214-fold. SDS-polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approx. 29 kDa. The four pesticides dose-dependently inhibited in vitro CA activity. IC₅₀ values for deltamethrin, diazinon, propoxur and cypermethrin were 0.137, 0.267, 0.420 and 0.460 μM, respectively. In vitro results showed that pesticides inhibit rtCA activity with rank order of deltamethrin > diazinon > propoxur > cypermethrin. Besides, in vivo studies of deltamethrin were performed on CA activity of rainbow trout gill. rtCA was significantly inhibited at three concentrations (0.25, 1.0 and 2.5 μg/L) at 24 and 48 h.     

Glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) enzyme activities in different tissues of Sarotherodon mossambicus (Peters) exposed to a carbamate pesticide,carbaryl

The activity levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) enzymes were estimated in liver, muscle and brain tissues of the fish, Sarotherodon mossambicus (Peters), which had been exposed to sub-lethal (3 mg litre⁻¹) and lethal (25 mg litre⁻¹) concentrations of the carbamate insecticide carbaryl. Based on the results obtained, the changes in GOT and GPT levels in liver, muscle and brain following different periods of sub-lethal and lethal carbaryl exposures suggested that S mossambicus showed adaptive elevation in the activity levels of the two aminotransferase enzymes in the tissues, thereby probably aiding gluconeogenesis through transamination of glucogenic amino acids to meet the energy demand under carbaryl toxicity. © 1999 Society of Chemical Industry     

Glucose metabolism in hepatopancreas and gill of Lamellidens marginallis during methyl parathion toxicity

Changes in glucose metabolism were studied in hepatopancreas and gill of freshwater mussel, Lamellidens marginalis, exposed to a sublethal concentration (8 ppm) of methyl parathion. A slight decrease in glycogen and pyruvate and an increase in lactate levels were observed. An increase in phosphorylase and aldolase suggested increased formation of trioses during methyl parathion toxicity. The decrease in lactate dehydrogenase activity and increase in lactate content indicated reduced mobilization of pyruvate into the citric acid cycle. Glucose-6-phosphate dehydrogenase activity was increased, suggesting enhanced oxidation of glucose by the HMP shunt pathway. Citric acid cycle enzymes such as isocitrate, succinate, and malate dehydrogenases were found to be decreased, suggesting abnormality in mitochondrial oxidative metabolism as a consequence of methyl parathion toxicity. The decreased cytochrome c oxidase and Mg²⁺-ATPase, apart from citric acid cycle enzymes, indicated impaired energy synthesis as a result of reduced aerobic oxidation of glycose. The increase in acid and alkaline phosphatase activities suggested enhanced breakdown of phosphate to release energy in view of inhibition of the ATPase system during methyl parathion stress. The changes were more pronounced in hepatopancreas as compared to gill of mussel exposed to methyl parathion.     

Activity changes of glyoxalase system enzymes and glutathione-S-transferase in the bivalve mollusc Scapharca inaequivalvis exposed to the organophosphate chlorpyrifos

The present report was aimed to study possible changes in the activity of glutathione-dependent enzymes glyoxalase I (G I) and glyoxalase II (G II), forming the glyoxalase system, and glutathione-S-transferase in tissues (gills, foot, and digestive gland) of the mediterranean bivalve mollusc Scapharca inaequivalvis, after a 4- or 15-day exposure to a sublethal concentration (0.1 ppm) of the organophosphate pesticide chlorpyrifos (CPF). Control experiments were also performed on untreated animals. G I activity decreased in gills and foot, and increased in the digestive gland after 4 days of CPF exposure. Glyoxalase II activity in the gills decreased after a 15 days exposure to CPF, was unchanged in the foot and increased after 15 days in the digestive gland. Glutathione-S-transferase activity, which is involved in the detoxification of xenobiotics, increased significantly in all tissues after CPF exposure. The results suggest that sublethal CPF exposure of S. inaequivalvis does not compromise the cellular redox balance in these tissues.