Sublethal Concentrations Of Chlorpyrifos Research Articles

Effect of chlorpyrifos-exposure on the expression levels of CYP genes in Daphnia magna and examination of a possibility that an up-regulated clan 3 CYP, CYP360A8, reacts with pesticides.

Daphnia magna is a test organism used for ecological risk assessments of pesticides, but little is known about the expression levels of cytochrome P450s (CYP)s and their changes after pesticide exposure in the less than 24-h-olds used for ecotoxicity tests. In this study, D. magna juveniles were exposed to 0.2 μg/L of chlorpyrifos under the conditions for acute immobilization test as specified by the OECD test guideline for 24 h, and then the gene expression was compared between the control and chlorpyrifos-exposure groups by RNA-sequencing analysis, with a focus on CYP genes. Among 38 CYP genes expressed in the control group, seven were significantly up-regulated while two were significantly down-regulated in the chlorpyrifos-exposure group. Although the sublethal concentration of chlorpyrifos did not change their expression levels so drastically (0.8 < fold change < 2.6), CY360A8 of D. magna (DmCYP360A8), which had been proposed to be responsible for metabolism of xenobiotics, was abundantly expressed in controls yet up-regulated by chlorpyrifos. Therefore, homology modeling of DmCYP360A8 was performed based on the amino acid sequence, and then molecular docking simulations with the insecticides that were indicated to be metabolized by CYPs in D. magna were conducted. The results indicated that DmCYP360A8 could contribute to the metabolism of diazinon and chlorfenapyr but not chlorpyrifos. These findings suggest that chlorpyrifos is probably detoxified by other CYP(s) including up-regulated and/or constitutively expressed one(s).

HISTOPATHOLOGICAL EFFECT OF CHLORPYRIFOS PESTICIDE ON LIVER AND KIDNEY OF FRESH WATER FISH CATLA CATLA

All the natural water bodies are continuously being contaminated with toxic chemicals from industrial, agricultural and domestic activities. Chlorpyrifos is one of the most widely used organophosphate pesticides. Excessive application of pesticides from agricultural fields contaminate aquatic medium, resulting in serious damage to non-target species, including fish. The fish as a bioindicator of aquatic medium plays an important role in the monitoring of water pollution. Histopathology is an important tool for evaluating the action of any toxicant at tissue level. The focus of the present study is to measure the histological changes in liver and kidney of the fresh water fish Catla catla exposed to sub lethal concentrations of chlorpyrifos 1/15th of the 96 hour LC50 values for the period of 10 and 20 days. The fish exposed to chlorpyrifos showed mild vacuolar hepatocytes, dilation of central vein, vaculor degenerative changes, mild hyperaemia and severe hyperaemia of central vein in the liver tissue slide. The changes observed in kidney slide due to the impact of chlorpyrifos toxicity are hemorrhage, degenerative changes in between tubules, constriction of primary tubule, vacuolardegenerative changes, necrosis in epithelial lining of proximal tubules, chlorpyrifos compound induced.

Investigation of oxidative stress enzymes and histological alterations in tilapia exposed to chlorpyrifos.

Chlorpyrifos (ChF) is an organophosphate pesticide that is widely used in agricultural fields and indoor for controlling pests. Aquatic ecosystems are the recipients of various pesticide residues due to leaching spray drift and agricultural runoff and pose toxicity for aquatic organisms. Therefore, the current study was designed to investigate the oxidative stress enzymes and histological alterations in the vital organs of tilapia due to ChF exposure. LC50 (24h) was calculated as 52.78μg/l by exposing tilapia with different acute concentrations of ChF. For assessment of sub-lethal toxicity of ChF, the fish were divided into four groups (ChF1, ChF2, ChF3, and control group). ChF1 group was treated with 1/15th of LC50, whereas ChF2 and ChF3 groups were treated with 1/10th and 1/5th of LC50, respectively for 14days. After that, ChF induced changes in oxidative stress enzymes and histological alterations were evaluated. It was found that the level of glutathione S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) increased significantly in the liver of ChF-treated tilapia. Histological study of liver tissues showed an increased number of Kupffer cells, hydropic degeneration, necrosis, and hemorrhage. In the spleen of treated fish, increased melanomacrophage centers, necrosis, and congestion were detected. Disorganized muscle fibers, cardiac muscle fiber degeneration, and coagulative necrosis were observed in the heart of ChF-treated fish. It is concluded that sub-lethal concentrations of ChF can induce oxidative stress and histological alterations in the tissues of tilapia.

Comparative toxicity of chlorpyrifos: Sublethal effects on enzyme activities and histopathology of Mugil cephalus and Chanos chanos

Ecotoxicological data and potential impact of chlorpyrifos (CPF) in the region are scarce for prescribing safety limits. Therefore, toxicity and sublethal impact of CPF on fish fingerlings of Mugil cephalus (3.0 ± 1.2 cm) and Chanos chanos (3.0 ± 1.5 cm) were studied. Acute and chronic toxicity tests were conducted by continuous flow through method and derived 96 h median lethal concentration (LC50). Mean LC50 value of 1.13 μg/L for M. cephalus, and 3.20 μg/L for C. chanos were derived by Probit. Chronic toxicity tests were conducted for 30 days and determined no observed effect concentration values of 0.09 μg/L 0.17 μg/L and lowest observed effect concentration values of 0.16 μg/L 0.32 μg/L and chronic values of 0.13 μg/L 0.25 μg/L for M. cephalus and C. chanos respectively. Key biomarker enzyme activities viz., EST, SOD and MDH were studied at sublethal concentrations of CPF. Native gel electrophoresis revealed gradual decrease in isoforms of EST and SOD activities, whereas MDH activity increased in fingerlings. These responses indicate inhibition of cholinesterase, antioxidants and synthesis of ATPs in the cells due to CPF stress. Pathological lesions were evaluated in gill and eye tissues of fingerlings. Epithelial fusion and degenerative changes were prominent in primary lamellae. Hyperplasia, lifting epithelium, fusion of lamellae and necrosis were evidenced in the secondary lamellae. Cellular anomalies in the retina of the eye of C. chanos include vacuoles in nerve fiber layer, shrinkage of outer plexiform layer and detachment of pigment epithelium layer. These changes indicate physiological disturbance in the gill and eye.

Effect of Tunceli garlic on some immunological parameters in Cyprinus carpio exposed to chlorpyrifos

In this study, the effect of Tunceli garlic (Allium tuncelianium) on hemoglobin (Hb) level, oxidative radical production of neutrophils (Nitoblue tetrazolium assay-NBT activity) and total immunoglobulin (TI) content in carp (Cyprinus carpio) exposed to chlorpyrifos (CPF) was investigated. The 96 hour LC50 value of CPF on C. carpio was calculated to as 0.230 mg/L. The fishes were exposed to sublethal concentration of chlorpyrifos (1/8 of LC50 value: 0.029 mg/L), and Tunceli garlic (20 and 40 g/kg diet) was simultaneously administered for 21 days. Blood samples were taken from the fishes at 7, 14 and 21 days and analysed to determine the Hb levels, the NBT activity and the TI content. There was a significant decrease in the Hb level, the NBT activity and the TI content of CPF-treated fish. However, Tunceli garlic reversed the Hb level, the NBT activity and the TI content. In conclusion, this study demonstrated that CPF had a negative effect on the immunological values of the fish. The simultaneous administration of Tunceli garlic was neutralised CPF-induced toxicity.

Chlorpyrifos-induced biochemical changes in Cyprinus carpio: Ameliorative effect of curcumin

The aim of this study was to determine protective effects of curcumin on some haematological values and oxidant/antioxidant status in Cyprinus carpio exposed to chlorpyrifos. The fish were exposed to two sublethal concentrations of chlorpyrifos (0.040 and 0.080mgL), and curcumin (100mg per kg of fish weight) was simultaneously administered for 14 days. Blood and tissue (liver, kidney, and gill) samples were collected at the end of the experiment and analysed to determine the haematological profile (red blood cell count, white blood cell count, haemoglobin concentration, and haematocrit level) and oxidant/antioxidant status (malondialdehyde level and superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase activities) of the fish. There was a significant decrease in the red blood cell count, the haemoglobin concentration, and the haematocrit level and a increase in the white blood cell count of CPF-treated fish. The results revealed a significant increase in the malondialdehyde levels of the groups that were exposed to CPF. Conversely, the MDA levels were significantly decreased by curcumin. Also, CPF exposure caused a significant increase in the superoxide dismutase and glutathione-S-transferase activities and a significant decrease in the catalase and glutathione peroxidase activities. However, curcumin reversed the superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase activities. In conclusion, this study demonstrated that CPF had a negative effect on the haematological values and the oxidant/antioxidant status of the fish. The simultaneous administration of curcumin was neutralised CPF-induced toxicity.

Biomarker and histopathological responses of Lates calcarifer on exposure to sub lethal concentrations of chlorpyrifos

Bioassay tests on fingerlings (3.0 ± 0.5cm) of euryhyaline fish Lates calcarifer were conducted using customized continuous flow through system and derived 96h acute toxicity value for chlorpyrifos (CPF). Based on the measured concentration of CPF mean median lethal concentration (LC50) of 1.07µg/l with lower and upper 95% confidence limits (0.95 and 1.19µg/l). No observed Effect Concentration (NOEC), Lowest Observed Effect Concentration (LOEC) and chronic values were found to be 0.4, 0.09 and 0.07µg/l respectively. Key biomarker enzyme activities such as esterase, superoxide dismutase and malate dehydrogenase were measured in whole body tissues of the fish fingerlings on exposure to sublethal toxicity of CPF resulting in inhibition of enzyme activities. Native gel electrophoresis revealed single isoform of SOD and MDH enzyme activities exhibiting time and concentration dependent inhibition. Interestingly, three isoforms of esterase activity were witnessed, two isoforms didn’t show changes and one isoform was completely inhibited. The observed changes indicated continuous production of reactive oxygen species (ROS) in cells, affecting the integrity and function of cell membrane. Decreased MDH activity indicates reduction of ATP production in the mitochondria leading to susceptibility of fish fingerlings due to the imposed CPF toxicity. Histopathological changes are evident as physiological signatures of chemical interactions in the cell and are prominently used for the evaluation of toxic effects. Gills and eye tissues were selected considering the possible effects on respiratory surfaces and vision impairment. Their tissue sections were observed for changes in primary & secondary lamellae, and retina of the eye respectively. Prominent pathological lesions of gills and retina of the eye include degeneration of cells, fusion, lifting of epithelium and increased cellular space, detachment of pigment epithelium, fusion of photoreceptor cells, respectively on exposure to 30 days of sub lethal concentrations. CPF was found to be highly toxic, affecting the vital functions of respiration, vision and cellular activities leading to susceptibility of fish fingerlings.

Genotoxic and hematological effects of chlorpyrifos exposure on freshwater fish Labeo rohita

Chlorpyrifos is a commonly used organophosphate insecticide that causes toxicological effects in aquatic organisms especially in fish. This study determined the effects of chlorpyrifos on the genotoxic and hematological parameters of freshwater fish, Labeo rohita. The genotoxic effects of different sublethal concentrations of chlorpyrifos were investigated in the erythrocytes of Labeo rohita (commonly known as Rohu) using the Micronucleus test. Effects of chlorpyrifos on the hematological parameters of the fish were also observed. Fish specimens were exposed to three sublethal concentrations of chlorpyrifos viz., sublethal I (SL-I, 1/6th of LC50 = ∼73.8 μg/L), sublethal II (SL-II, 1/4th of LC50 = ∼110.7 μg/L) and sublethal III (SL-III, 1/2nd of LC50 = ∼221.4 μg/L) for 96 h. Blood samples were collected at every 24 h and were subjected to the Micronucleus assay. The observed micronucleus frequencies were concentration and time-dependent. The MN induction was significantly highest (p < 0.01) at all the concentrations on 96-h exposure. During the experimental period, hematological parameters like total erythrocytes count (TEC), hemoglobin (Hb) and packed cell volume (PCV) decreased, whereas total leukocytes count (TLC) increased. This study indicated that the Micronucleus assay is a useful tool to detect genotoxic potential of chlorpyrifos in fish.

Effect of Insecticide "Chlorpyrifos" on Immune Response of Oreochromis niloticus

This study was carried out to determine the level of Chlorpyrifos (CPF), an organophosphorus insecticide, in water of Abbassa and Sahl El-Hussinia fish farms and investigate the sub lethal concentration of this pesticide on immune response of Oreochromis niloticus ( O. niloticus ) . Water samples were collected from Abbassa and Sahl El-Hussinia fish farms and analyzed using Gas chromatograph (GC) for detection of Chlorpyrifos level. Two hundred and ninety O. niloticus with average body weight (35±0.5 g) were collected from Abbassa fish hatchery, Sharkia Governorate, Egypt. One hundred and seventy of these fish were used for the determination of acute toxicity of Chlorpyrifos, while, another 120 fish were used for the determination of the effect of different sub lethal concentrations of Chlorpyrifos (1/8, 1/20 and 1/43 of 96 h LC 50 ) on the immune response of O. niloticus . It was found that; the levels of Chlorpyrifos in Abbassa and Sahl El-Hussinia fish farms were 0.008 and 0.0016 mg/L, respectively. The LC 50 value of Chlorpyrifos was 0.07mg/L. Sub-lethal concentration of Chlorpyrifos altered the non-specific immunological parameters (namely, total globulin, immunoglobulin M (IgM), lysozyme, nitric oxide, phagocytic activity) and interlukin-1β (IL-1β) of O. niloticus . It could be concluded that water of Abbassa and Sahl El-Hussinia fish farms have had detectable residue levels of CPF, which was altered the immunological status of O. niloticus .

Lethal and Demographic Impact of Chlorpyrifos and Spinosad on the Ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae).

The appropriate use of biological agents and chemical compounds is necessary to establish successful integrated pest management (IPM) programs. Thus, the off-target effects of pesticides on biological control agents are essential considerations of IPM. In this study, the effects of lethal and sublethal concentrations of chlorpyrifos and spinosad on the demographic parameters of Habrobracon hebetor (Say) (Hymenoptera: Braconidae) were assessed. Bioassays were carried out on immature and adult stages by using dipping and contact exposure of dry pesticide residue on an inert material, respectively. The lethal concentration (LC)50 values of chlorpyrifos and spinosad were 3.69 and 151.37 ppm, respectively, on the larval stage and 1.75 and 117.37 ppm, respectively, on adults. Hazard quotient (HQ) values for chlorpyrifos and spinosad were 400 and 2.2, respectively, on the larval stage and 857.14 and 2.84, respectively, on adults. A low lethal concentration (LC30) was used to assess the sublethal effects of both pesticides on the surviving females. In each treatment, 25 survivors were randomly selected and transferred into 6-cm Petri dishes. Adults were provided daily with last instars of Anagasta kuehniella (Zeller) as a host until all of the females died. The number of eggs laid, percent of larvae hatched, longevity, and sex ratio were recorded. Stable population growth parameters were estimated by the Jackknife method. In control, chlorpyrifos, and spinosad treatments, the intrinsic rates of increase (r m) values were 0.23, 0.10, and 0.21, respectively. The results of this study suggest a relative compatibility between spinosad use and H. hebetor. Finally, further studies should be conducted under natural conditions to verify the compatibility of spinosad with H. hebetor in IPM programs.

Effects of Chlorpyrifos and Endosulfan on Different Life Stages of the Freshwater Burrowing Crab Zilchiopsis collastinensis P.: Protective Role of Chorion

The lethality (LC50) of endosulfan and chlorpyrifos is higher in adults (1990 and 156.78 μg/L respectively) than in embryos (95380 and 1242.54 μg/L respectively) of the crab Zilchiopsis collastinensis. The thickened embryonic coat of the freshwater species might isolate the embryo inside the egg, reducing the toxicity. Sublethal concentrations of chlorpyrifos and endosulfan caused an increase in hatching time and a decrease in effective hatching (p < 0.05), and only the control crabs survived until the first molt cycle. The effects of long-term exposure should be evaluated in the offspring in addition to the acute toxicity.

Assessment of Potential Sublethal Effects of Various Insecticides on Key Biological Traits of The Tobacco Whitefly, Bemisia tabaci

The tobacco whitefly Bemisia tabaci is one of the most devastating pests worldwide. Current management of B. tabaci relies upon the frequent applications of insecticides. In addition to direct mortality by typical acute toxicity (lethal effect), insecticides may also impair various key biological traits of the exposed insects through physiological and behavioral sublethal effects. Identifying and characterizing such effects could be crucial for understanding the global effects of insecticides on the pest and therefore for optimizing its management in the crops. We assessed the effects of sublethal and low-lethal concentrations of four widely used insecticides on the fecundity, honeydew excretion and feeding behavior of B. tabaci adults. The probing activity of the whiteflies feeding on treated cotton seedlings was recorded by an Electrical Penetration Graph (EPG). The results showed that imidacloprid and bifenthrin caused a reduction in phloem feeding even at sublethal concentrations. In addition, the honeydew excretions and fecundity levels of adults feeding on leaf discs treated with these concentrations were significantly lower than the untreated ones. While, sublethal concentrations of chlorpyrifos and carbosulfan did not affect feeding behavior, honeydew excretion and fecundity of the whitefly. We demonstrated an antifeedant effect of the imidacloprid and bifenthrin on B. tabaci, whereas behavioral changes in adults feeding on leaves treated with chlorpyrifos and carbosulfan were more likely caused by the direct effects of the insecticides on the insects' nervous system itself. Our results show that aside from the lethal effect, the sublethal concentration of imidacloprid and bifenthrin impairs the phloem feeding, i.e. the most important feeding trait in a plant protection perspective. Indeed, this antifeedant property would give these insecticides potential to control insect pests indirectly. Therefore, the behavioral effects of sublethal concentrations of imidacloprid and bifenthrin may play an important role in the control of whitefly pests by increasing the toxicity persistence in treated crops.

Histopathological alterations in gill, liver and kidney of common carp exposed to chlorpyrifos

Histopathological alterations in gill, liver and kidney of common carp, Cyprinus carpio, intoxicated with sub-lethal concentrations of chlorpyrifos (O,O,-diethyl-O-3,5,6-trichloro-2-pyridyl phosphorothioate) pesticide (1 and 100 μg/L) for a period of 14 days were analyzed under light microscope. Gill exhibited hyperplasia and hypertrophy of gill epithelium, blood congestion, dilation of marginal channel, epithelial lifting, lamellar fusion, lamellar disorganization, lamellar aneurysm, rupture of the lamellar epithelium, rupture of pillar cells and necrosis. Alterations in hepatocytes were more pronounced, including nuclear and cellular hypertrophy, cellular atrophy, irregular contour of cells and nucleus, cytoplasmic vacuolation, cytoplasmic and nuclear degeneration, cellular rupture, pyknotic nucleus, necrosis and melanomacrophages aggregations. Histopathological lesions in kidney were cellular and nuclear hypertrophy, narrowing of tubular lumen, cytoplasmic vacuolation, hyaline droplet degeneration, nuclear degeneration, occlusion of tubular lumen, tubular regeneration, dilation of glomerular capillaries, degeneration of glomerulus and hemorrhage in Bowman's space. The most significant conclusion drawn from this study was that with the increased concentration and duration the toxicosis of chlorpyrifos would be enhanced as shown through the analysis of mean assessment value (MAV) and degree of tissue changes (DTC) also.

Sub-lethal effect of chlorpyrifos on protein metabolism of the food fish Clarias batrachus and monitoring of recovery

The effect of sub-lethal concentrations of chlorpyrifos on protein metabolism in physiological important tissues, namely gills, kidney, liver, and muscle of the freshwater fish, Clarias batrachus, was studied. Fish were exposed to 1/20th and 1/10th of 96 h LC50 concentrations for 7, 14, 21, and 28 days. After 28 days of exposure, fish were released into fresh water and kept in the same for 21 days in order to study the recovery. Fish were sacrificed at the stipulated periods and gills, kidney, liver, and muscle tissues were used for the estimation of total protein, amino acids, ammonia, urea, glutamine, protease, transaminases, and phosphatases. Total protein, amino acid, and ammonia contents were decreased in all tissues for 28 days and recovery was observed during the recovery period. Urea and glutamine levels were elevated, except in kidneys, and recovered at the end of the recovery period. The activities of protease, alanine, and aspartate aminotransferases, and acid and alkaline phosphatases were elevated in the tissues for 28 days exposure at both concentrations. Recovery of these enzymes activities was noticed during depuration.

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.

Lysosomal neutral red retention time as a biomarker of organophosphate exposure in the earthworm Aporrectodea caliginosa: laboratory and semi-field experiments

Aporrectodea caliginosa is the most common endogeic (topsoil) earthworm in New Zealand and, because of its habitat, is potentially vulnerable to surface-applied pesticides. Lysosomal damage to earthworms, which can be visualized by the use of the neutral red retention assay (NRRA) has been evaluated in this species as a biomarker of organophosphate exposure. Earthworms were exposed in the laboratory to sub-lethal concentrations of chlorpyrifos and diazinon. In a semi-field experiment, earthworms were placed in mesocosms in a field sprayed with these pesticides at the rate recommended for a vegetable crop. In the laboratory, the neutral red retention time (NRRT) was significantly reduced following exposure to both pesticides compared with controls. In the semi-field experiment, earthworm NRRT was significantly reduced by both pesticides. These experiments have shown that the NRRA is very sensitive to exposure to chlorpyrifos and diazinon even at field rates. It therefore shows promise as a potential biomarker of contamination of soil by organophosphates.

Sublethal effects of insecticides on adult longevity and fecundity of German cockroaches (Dictyoptera: Blattellidae).

The effects of sublethal concentrations of chlorpyrifos, cyfluthrin, and hydramethylnon on adult longevity and fecundity of German cockroaches, Blattella germanica (L.), were investigated. Longevity of males declined linearly with increasing doses of insecticide. An LD50 of cyfluthrin decreased male longevity by 52%, whereas an LD50 of hydramethylnon reduced male longevity by 81%. Longevity of females increased linearly with increasing sublethal doses of chlorpyrifos, whereas all doses of both cyfluthrin and hydramethylnon decreased longevity of females. Fecundity increased linearly with increasing sublethal doses of chlorpyrifos. Number of oothecae formed, oothecae hatched, and number of offspring produced in each ootheca increased with increasing sublethal concentrations of chlorpyrifos. In contrast, fecundity declined with increasing sublethal concentrations of cyfluthrin and hydramethylnon. Formation of the first ootheca occurred approximately 8 d after mating for untreated females but generally longer with sublethal concentrations of all insecticides. The period between oothecae hatch and the formation of subsequent oothecae increased with successive oothecae in all treatments.