Some hematological,biochemical, and enzymological parameters of a fresh-water teleost fish, Channa punctatus, exposed to sublethal concentrations of quinalphos

Alterations in the levels of hemoglobin, total plasma proteins, glucose, and lactic acid in the blood; glycogen and lactic acid content of liver and white skeletal muscle; and the activities of lactate dehydrogenase, pyruvate dehydrogenase, and succinate dehydrogenase in liver, kidney, intestine, brain, gills, and muscles were examined in the fresh-water snake-head fish, Channa punctatus, after exposure to a sublethal concentration (25 μg/liter) of quinalphos for 60 and 120 days. Hemoglobin, plasma protein, glucose, and lactic acid decreased in pesticide-exposed fish. The glycogen content of the liver and muscles increased but lactic acid decreased. Lactate dehydrogenase activity decreased in all six tissues. Pyruvate dehydrogenase activity of liver, kidney, gill, and muscle decreased, but the enzyme activity was elevated in intestine and brain. In intestine, succinate dehydrogenase activity was elevated, and in the remaining five tissues the enzyme activity was significantly reduced. The present study showed that formation of glycogen and its breakdown was impaired in the liver, and aerobic oxidation of nutrients was adversely affected in quinalphos-exposed fish.     

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.     

Interactive effect of monocrotophos and ammonium chloride on the freshwater fish Oreochromis mossambicus with reference to lactate/pyruvate ratio

Sublethal effect of monocrotophos (pesticide) and ammonium chloride (fertilizer) was studied in the freshwater fish Oreochromis mossambicus, with reference to carbohydrate metabolism for a period of 96 h. The glycogen content was analysed in liver and muscle, while the lactate and pyruvate were assessed in blood along with liver and muscle. The results revealed that the glycogen content was found to be decreasing. In contrast increase in the tissues lactate and pyruvate level was found in the fishes exposed to pesticide and fertilizer individually and in combinations. The combined effects of these chemicals were more toxic to carbohydrate metabolism than the effect produced by the individual chemicals. The results were tested to search for statistical significance. The calculated lactate and pyruvate ratio (L/P) indicated that the fishes were under chemical stress.     

Effect of a paired mixture of aldrin and formothion on carbohydrate metabolism in a fish, Heteropneustes fossilis

Exposure of the Indian catfish (Heteropneustes fossilis) to a high sublethal concentration, 0.114 ppm (0.8 of the 96-hr LC₅₀) of a mixture of aldrin and formothion, added in a one-to-one ratio, for 3, 6, 12, 48, and 96 hr affected carbohydrate metabolism. Muscle glycogen decreased significantly at 6 and 48 hr; hepatic glycogen content declined at 3, 6, 12, and 48 hr in exposed fish. Blood glucose levels in fish were significantly decreased at 3, 12, 48, and 96 hr. Mean value of blood pyruvate was significantly depressed at 3 hr. Blood lactate levels were elevated at 3, 6, 12, and 96 hr in pesticide-treated fish. The observed effects of the mixture of aldrin and formothion on carbohydrate metabolism in fish could prove useful as a rapid method for evaluating toxicity of pesticides and other toxicants.     

Concentration related responses of chlorpyriphos in antioxidant, anaerobic and protein synthesizing machinery of the freshwater fish, Heteropneustes fossilis

Impacts of chlorpyriphos (2, 4, 6 ppm) on the profiles of antioxidant (catalase) and anaerobic (lactate dehydrogenase) enzymes and other macromolecular contents (DNA, RNA, protein) of various tissues of the freshwater fish, Heteropneustes fossilis were studied. Chlorpyriphos significantly decreased the specific activity of catalase (CAT) and lactate dehydrogenase (LDH) in the brain, liver, gill and skeletal muscle of the fish. The reduction in specific activity might be due to the binding of chlorpyriphos or its metabolites with the enzyme molecules or affecting the synthesis and/or degradation of the enzymes. Like enzymes, the DNA, RNA and protein contents decreased in the brain, liver, gill and skeletal muscle of the fish as a function of increase in chlorpyriphos concentrations (2-6 ppm). The chlorpyriphos-induced reduction in these biochemical constituents might be because of alteration in their turnover (synthesis/degradation) in different tissues. The maximum effects on CAT, LDH, DNA, RNA and protein were obtained in response to 6 ppm chlorpyriphos. The present findings suggest chlorpyriphos concentration related impairment in antioxidative, anaerobic and protein synthesizing capacity of the fish. Therefore, the use of higher concentrations of chlorpyriphos should be avoided to protect the health of economically important freshwater food fish.     

Metabolism and absorption of methyl parathion by tobacco budworms resistant or susceptible to organophosphorus insecticides

Factors involved in insecticide resistance were evaluated by using ¹⁴C-labeled methyl parathion and aldrin to compare rates of absorption and metabolism by Heliothis virescens (F.) larvae that were resistant (R) and susceptible (S) to methyl parathion. Tests with third-stage R and S larvae suggested that the rate of insecticide absorption from the cuticular surface was not a major resistance factor. Further evidence for this assumption was demonstrated by the resistance of R larvae to injected and orally administered doses of methyl parathion. Smaller amounts of unmetabolized methyl parathion and aldrin were recovered from S larvae, an indication that differences in metabolism were probably related to the resistance.     

Organophosphate degradation by insecticide-resistant and susceptible populations of mosquitofish (Gambusia affinis)

Phosphorothionate and phosphate degradation was investigated as a factor which could influence the tolerance of organochlorine compound-resistant and susceptible mosquitofish (Gambusia affinis) to parathion and methyl parathion. The greater toxicity of methyl parathion than parathion can be attributed in part to a higher rate of degradation of methyl paraoxon than paraoxon (7-fold), but not to any difference in phosphorothionate dearylation. Resistant fish possess higher levels of microsomal mixed-function oxidases which can degrade methyl parathion (1.3-fold); these higher levels could contribute to the increased methyl parathion tolerance by this population over the susceptible population. Environmentally induced tolerance to parathion in the resistant population may be the result of increased levels of parathion degradation by induced mixed-function oxidases which can dearylate parathion. The increased tolerance of either insecticide by the resistant population is not caused by degradation of the phosphates by phosphotriesterases.     

Comparative toxicity,absorption, and metabolism of chlorpyrifos and its dimethyl homologue in methyl parathion-resistant and -susceptible tobacco budworms

Chlorpyrifos (Dowco 179) and its dimethyl homologue, chlorpyrifosmethyl (Dowco 214), were used to study the influence of the O,O-dialkyl group of organophosphorus insecticides on toxicity, absorption, and metabolism among larvae of the tobacco budworm [Heliothis virescens (F.)] from strains that were resistant (R) and susceptible (S) to methyl parathion. In toxicity tests, chlorpyrifos and chlorpyrifosmethyl were more toxic than methyl parathion to 3rd-stage R larvae but less toxic to S larvae. Chlorpyrifosmethyl was more toxic (3–4 ×) than chlorpyrifos to both strains of larvae, and the results of absorption studies indicated that the toxicity differential of the homologues may be explained in part by the more rapid absorption of the dimethyl form. Studies of the in vivo metabolism of both Dowco compounds indicated that each was degraded mainly by the cleavage of the pyridylphosphate linkage. In vitro tests demonstrated that the NADPH-dependent microsomal oxidases were of primary importance in detoxification, while glutathione (GSH)-dependent mechanisms (aryl- and alkyltransferases) present in the soluble cell fractions were of lesser importance. O-dealkylation occurred only with chlorpyrifosmethyl. The R larvae demonstrated greater capability in detoxifying both compounds in the comparative in vivo and in vitro studies of metabolism, but the differences were more apparent during the 5th instar than during the 3rd instar.     

The mechanism of fungistatic action of sec-butylamine: II. The effect of sec-butylamine on pyruvate oxidation by mitochondria of Penicillium digitatum and on the pyruvate dehydrogenase complex

sec-Butylamine at 5 mM inhibited the oxidation of pyruvate by mitochondria isolated from hyphae of Penicillium digitalum, but had little effect on the oxidation of citrate, isocitrate, succinate, malate, acetyl-coenzyme A, or reduced nicotinamide adenine dinucleotide. sec-Butylamine did not interfere with oxidative phosphorylation, as evidenced by similar $\text{P}\text{O}$ ratios in treated and control mitochondria. The pyruvate dehydrogenase complex (EC 1.2.4.1) isolated from young hyphae of P. digitatum was inhibited strongly by 20 mM sec-butylamine, whereas other tricarboxylic acid cycle enzymes were only slightly affected at most. Inhibition of the pyruvate dehydrogenase complex by sec-butylamine was competitive with respect to pyruvate. The K_i for sec-butylamine in the reaction was 1.38 × 10^?2M, and the K_m for pyruvate was 2.28 × 10^?4M. These observations and other evidence derived from studies with intact hyphae support the hypothesis that the pyruvate dehydrogenase complex is the primary site of the fungistatic action of sec-butylamine.     

群居型和散居型亚洲小车蝗成虫能量代谢相关酶活性测定

为明确亚洲小车蝗Oedaleus asiaticus成虫飞行肌对其体内能源物质利用的情况,采用室内生化法测定能源物质代谢相关5种酶,即3-磷酸甘油醛脱氢酶(glyceraldehyde phosphate dehydrogenase,GAPDH)、3-磷酸甘油脱氢酶(glycerol-3-phosphate dehyd...     

Toxicity in the brain of organophosphate insecticides: Comparison of the toxicities of metabolites with parent compounds using an intracerebral injection method

The toxicity in the brain of several parathion, fenthion, and fensulfothion insecticides and their toxic metabolites was determined by a technique of directly injecting the compounds into the region of the third ventricle of conscious mice, an area rich in cholinesterase activity. The results were compared on a body weight basis to the toxicity of these compounds when given by ip and oral routes. The results show that there is a direct relationship between the relative inhibition of cholinesterase activity in the brain by the organophosphates (e.g., methyl paraoxon, Sumioxon, and some members of the fenthion series) and the toxicity of these compounds in the brain. Methyl paraoxon and Sumioxon were found to be very toxic in the brain, Sumioxon being three to four times less toxic than methyl paraoxon. This is of the same order of effect of these compounds in inhibiting cholinesterases. It is concluded that any selective effects of Sumithion compared with methyl parathion must be due to the greater rate of metabolism of Sumithion to less toxic metabolites as well as to the lower toxicity of the oxon metabolite and not due to the relative rates of penetration of the toxic oxygen metabolites as previously suggested [J. Miyamoto, Agr. Biol. Chem.28, 422 (1964)]. A gas-liquid chromatographic method was employed to assess the distribution in the brain following intracerebral injection of the parathion-type compounds. The results suggest that there may be intracerebral metabolism of thionophosphates in vivo.     

Insect glutathione S-transferase: Separation of transferases from fat bodies of American cockroaches active on organophosphorus triesters

Several glutathione S-transferases which catalyze the conjugation of reduced glutathione with organophosphorus triesters were separated from fat bodies of adult female American cockroaches, Periplaneta americana (L.). Two transferases (I, V) were active on diazinon and three transferases (II, III, IV) were active on methyl parathion. The transferase (I) active on the pyrimidinyl moiety of diazinon was distinguishable from the other transferases on the O-methyl portion of methyl parathion, as shown by chromatographic properties, and additionally it was almost inactive or less active on 3,4-dichloronitrobenzene, methyl iodide, p-nitrobenzyl chloride, trans-cinnamaldehyde, and 1,2-epoxy-3-(p-nitrophenoxy)propane. Transferase II had high activities with “aryl” and “aralkyl” compounds, transferase III with “epoxide” and “alkene,” and transferase IV with “alkyl,” “aryl,” and “aralkyl” compounds. This indicated that the transferases had overlapping substrate specificities. The molecular weight was 35,000–37,000 for both of the enzymes active on methyl parathion and diazinon. The pH optima with methyl parathion and diazinon were about 8.5 and 6.5, respectively. At a glutathione concentration of 5 mM, Michaelis constants were 0.28 and 0.13 mM for methyl parathion and diazinon, respectively.     

Influence of Atrazine and Roundup pesticides on biochemical and molecular aspects of Biomphalaria alexandrina snails

The excessive use of pesticides in agriculture has sparkled the interest of scientists in investigating the harmful effects of these compounds. The present study evaluates the pesticides Atrazine and Roundup (glyphosate) on biochemical and molecular aspects of Biomphalaria alexandrina snails. The results showed that LC₁₀ of these two pesticides caused considerable reduction in survival rates and egg production of treated snails. Additionally, Atrazine proved to be more toxic to B. alexandrina snails than Roundup. In treated snails, glucose concentration (GL) in the hemolymph as well as lactate (LT) and free amino acid (FAA) in soft tissues of treated snails increased while glycogen (GN), pyruvate (PV), total protein (TP), nucleic acids (DNA and RNA) levels in snail’s tissues decreased. The activities of glycogen phosphorylase (GP), superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR), succinic dehydrogenase (SDH), acetylcholinesterase (AChE), lactic dehydrogenase (LDH) and phosphatases (ACP and ALP) enzymes in homogenate of snail’s tissues were reduced in response to the treatment with the two pesticides while lipid peroxide (LP) and transaminases (GOT and GPT) activity increased (P < 0.001). The changes in the number, position and intensity of DNA bands induced by pesticides may be attributed to the fact that pesticide can induce genotoxicity through DNA damage. It was concluded that the pollution of the aquatic environment by Atrazine and Roundup pesticides, would adversely affect the metabolism of the B. alexandrina snails, and have adverse effects on its reproduction.     

Persistent sub-lethal chlorine exposure elicits the temperature induced stress responses in Cyprinus carpio early fingerlings

Thermal effluents discharged through cooling systems of nuclear power plants often contain chlorine (used to control bio-fouling), which may affect the metabolic status of fishes. In order to evaluate the hypothesis, we tested the effect of high temperature and a persistent sub-lethal chlorine exposure on stress responses in Cyprinus carpio advanced fingerlings. Fishes were acclimated to four different temperatures (26, 31, 33, and 36 °C) and maintained for 30 days in two different groups. Subsequently, one of the groups was exposed to persistent chlorine (0.1 mg L₋₁) for another 28 days and was compared with their respective temperature controls (without chlorine exposure). Sub-lethal doses of pollutants and increasing temperatures with in the tolerance range may not always register any morphological changes Therefore, we studied organ specific biochemical pathways viz. aspartate amino transferase, alanine amino transferase (enzymes of protein metabolism) in liver and muscle; fructose 1,6 diphosphatase (gluconeogenic pathway), in liver; pyruvate kinase, malate dehydrogenase, and lactate dehydrogenase (glycolytic pathway) in muscle; glucose-6-phosphate dehydrogenase (pentose phosphate pathway) in liver; alkaline phosphatase (phosphorus metabolism) in intestine, liver, and muscle; acetylcholine esterase (neurotransmitting enzyme) in brain, and adenosine triphosphate (for membrane transport) in gills at two different acclimation periods (14 and 28 days). The results indicate that C. carpio fingerlings demonstrated metabolic readjustments with increasing temperatures, in order to cope with energy demand of the cell. However, exposure to chlorine at higher temperatures affected protein metabolism, gluconeogenic pathway and subsequently glycolytic pathway, leading to an energy-limited condition. In addition, alteration of membrane transport and neurotransmission might be an early indication of cellular damage. Overall results indicate that persistent sub-lethal chlorine exposure elicits temperature induced stress response in C. carpio early fingerlings.     

Comparative studies on carbofuran-induced changes in some cytoplasmic and mitochondrial enzymes and proteins of epigeic, anecic and endogeic earthworms

Exposure of epigeic (Perionyx sansibaricus), anecic (Lampito mauritii) and endogeic (Metaphire posthuma) earthworms to four different concentrations (10 ppm, 20 ppm, 40 ppm and 80 ppm) of carbofuran for 16 days induced specific activities of cytoplasmic malate dehydrogenase (cMDH), mitochondrial malate dehydrogenase (mMDH) and lactate dehydrogenase (LDH). P. sansibaricus showed 33-52% increase in specific activities of aerobic (cMDH, mMDH) and anaerobic (LDH) enzymes and proteins (cytoplasmic and mitochondrial). In L. mauritii, the enhancement in enzyme and protein level was 30-46%. Similarly, M. posthuma indicated 29-43% increase in profile of enzymes and proteins. The significant increase was observed on 2nd or 3rd day and it was maximum on 16th day of carbofuran exposure. Maximum effect of carbofuran was on epigeic earthworm. The sensitivity of mitochondrial dehydrogenase and protein of different earthworm species to carbofuran appears to be associated with oxygen availability in different strata of soil habitats. Inductions in enzyme specific activity and protein content were concentration and time-dependent. The effect was more pronounced on aerobic enzymes (cMDH, mMDH) as compared to anaerobic (LDH) one. The increases in mitochondrial enzyme (47%) and protein (43%) were more than the cytoplasmic enzymes (35%) and protein (31%), which suggest a greater effect of carbofuran on respiratory metabolism of earthworms. The carbofuran-dependent increase in cytoplasmic and mitochondrial enzymes and proteins might be due to increased synthesis of metabolic enzymes and stress proteins.     

Binding characteristics of methyl parathion to photosynthetic membranes of Chlorella

When methyl parathion is added at a given concentration to exponentially growing Chlorella cultures containing different numbers of cells, the inhibition of cell number, packed cell volume, pigment content, or photosynthesis has been found to be a function of the cell number, the inhibition being decreased with increased number of cells. Inhibition of photosynthesis has been studied further with a view to characterizing the mechanism of inhibition with a simple assumption that methyl parathion binds to a specific component in Chlorella cells to form an inhibitory complex. The concentration of methyl parathion causing 50% inhibition (I₅₀) of photosynthetic O₂ evolution increases linearly with increasing concentration of chlorophyll in the culture medium. With whole cells the inhibition constant (K_i) is 15 times greater than that with cell-free photosynthetic membranes. This shows that the cell wall acts as a permeability barrier. The relation between the I₅₀ and K_i values and the analyses of the Hill plot of the inhibition curves reveal one binding site per 0.5 chlorophyll molecule and a cooperative binding of methyl parathion with at least three binding sites per binding molecule. Mild treatment of the photosynthetic membranes with trypsin makes the photosynthetic electron transport insensitive to the insecticide, suggesting that the binding component is proteinaceous in nature and the binding sites are located on the external surface of the membrane. The reversal of methyl parathion inhibition is parallel to that of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (diuron) inhibition during trypsin treatment suggesting that the binding proteins for these two inhibitors are similar.     

Mechanism of fenvalerate-withdrawal-dependent recovery in metabolism of a catfish, Clarias batrachus

The effects of protein synthesis inhibitors on fenvalerate-withdrawal-dependent recovery in citrate synthase (CS), glucose 6-phosphate dehydrogenase (G6-PDH), lactate dehydrogenase (LDH), RNA, and protein of brain, liver, and skeletal muscle of Clarias batrachus were studied to explore possible mode of action of a pyrethroid insecticide at metabolic level. The administration of actinomycin D or cycloheximide inhibited partially or completely the withdrawal-dependent increase in enzyme activities, RNA, and protein contents, substantiating de novo synthesis of these macromolecules during recovery. The inhibition was more pronounced in response to cycloheximide reflecting recovery in translation process. This also suggests the possibility of fenvalerate-induced impairment of metabolism through inhibition of enzyme synthesis.     

Juvenile hormone analogs: Toxicity and cross-resistance in the housefly

The toxicity of several juvenile hormone analogs (JHAs) to susceptible and insecticide-resistant housefly (Musca domestica L.) strains was determined by an assay procedure in which larvae were exposed to residues of JHAs in glass vials. All JHAs tested were toxic and the most active compound, isopropyl 11-methoxy-3, 7, 11-trimethylododeca-2, 4-dienoate, was 100 times as toxic to the susceptible Orlando Regular strain as methyl parathion and 600 times as toxic as DDT.A 5- to 30-fold tolerance to the different JHAs was present in an insecticide resistant strain in which resistance is associated with a high level of NADPH-dependent microsomal oxidase activity controlled by a gene(s) on chromosome II. Cross-resistance was less marked in a strain with a chromosome V high oxidase gene and absent in strains with other resistance mechanisms.The data indicate that cross-resistance to JHAs in insects may occur in certain strains with high levels of oxidative detoxifying activity. Even so, the most active JHA was far more toxic to both susceptible and resistant strains than methyl parathion or DDT.     

Adult Drosophila melanogaster glutathione S-transferases: Effects of acute treatment with methyl parathion

The effect of acute treatment of methyl parathion (MP) on the expression of BSP/GSH-agarose purified glutathione S-transferases (GSTs) in Drosophila melanogaster was investigated. Using 2-D gel analysis of the identified Epsilon-class, only DmGSTE6 (100%) and DmGSTE7 (72%) demonstrated significant increases in expression, suggesting the possibility that both may be involved in MP metabolism. A smaller percentage increase was also observed in DmGSTD1 (18%), a known DDT dehydrochlorinase, DmGSTE3 and DmGSTE9 and a putative Epsilon-class GST, CG16936, were shown not to be responsive to the challenge. Our finding demonstrates that not all member of the Epsilon-class GST, which are known for their role in insecticide resistance, are immediately responsive to this toxic challenge.