Effect of vegetable oil factory effluent on the gonads of a freshwater teleost, Heteropneustes fossilis (Bl.) --a biochemical study

The effects of three sublethal concentrations (5,10,15% v/v) of vegetable oil factory effluent on biochemical changes in the gonads (ovaries/testes) of a freshwater teleost were studied. The results reveal a significant decline in the total lipid, cholesterol, phospholipid, and glycerine levels, but an increase in the free fatty acid levels, both in the ovaries and the testes of H. fossilis, following 150 days (mid-January through mid-June) of exposure to 10% and 15% concentrations of the effluent. This decrease/increase in the biochemical parameters was more or less dependent on the concentration of the effluent. The functional significance of any decrease/increase in the biochemical parameter(s) has been discussed.     

Biochemical study on the protective role of folic acid in rabbits treated with chromium (VI)

Deleterious effects of chromium (VI) compounds are diversified affecting almost all the organ systems in a wide variety of animals. Therefore, the present study was carried out to determine the effectiveness of folic acid (FA) in alleviating the toxicity of chromium (VI) on certain biochemical parameters, lipid peroxidation, and enzyme activities of male New Zealand white rabbits. Six rabbits per group were assigned to one of four treatment groups: 0 mg FA and 0 mg Cr(VI)/kg BW (control); 8.3 microg FA/kg BW; 5 mg Cr(VI)/kg BW; 5 mg Cr(VI) plus 8.3 microg FA/kg BW, respectively. Rabbits were orally administered their respective doses every day for 10 weeks. Results obtained showed that Cr(VI) significantly (P < 0.05) increased the levels of free radicals and the activity of glutathione S-transferase (GST), and decreased the content of sulfhydryl groups (SH groups) in liver, testes, brain, kidney, and lung. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AlP), acid phosphatase (AcP), and lactate dehydrogenase (LDH) were significantly decreased in liver and testes due to Cr(VI) administration. Also, AlP and AcP activities were significantly decreased in kidney and lung. The activity of acetylcholinesterase (AChE) was significantly decreased in brain and plasma. Contrariwise, the activities of AST and ALT were significantly increased in plasma, while AlP and AcP decreased. Chromium (VI) treatment caused a significant decrease in plasma total protein (TP) and globulin, and increased total lipids (TL), cholesterol, glucose, urea, creatinine, and bilirubin concentrations. Folic acid alone significantly decreased the levels of free radicals in liver, brain, and kidney, and increased the content of SH-group. The activities of AST, ALT, and LDH in liver; AST, ALT, AlP, AcP, and LDH in testes; AcP in kidney; AlP and AcP in lung, and LDH in brain were significantly increased. Plasma TP and albumin were increased, while urea and creatinine were decreased. The presence of FA with Cr(VI) restored the changes in enzyme activities and biochemical parameters. In conclusion, folic acid could be effective in the protection of chromium-induced toxicity.     

Dehydrogenase and phosphomonoesterase activities in groundnut (Arachis hypogaea L.) field after diazinon, imidacloprid and lindane treatments

Impacts of diazinon, imidacloprid and lindane treatments on dehydrogenase and alkaline phosphomonoesterase enzyme activities were determined in groundnut (Arachis hypogaea L.) field for three consecutive years (1997-1999). Diazinon was applied as both seed and soil treatments but imidacloprid and lindane were used for seed treatments only at recommended rates. Experiments were conducted at Agricultural Research Station Durgapura, Jaipur, Rajasthan, India. Diazinon residues were persist up till 60 days in both the cases. Average half-lives (t(1/2)) of diazinon were found 29.3 and 34.8 days, respectively, for seed and soil treatments. Diazinon seed treatment had no significant effect on dehydrogenase and alkaline phosphomonoesterase enzymes activities. In diazinon soil treatment, there were a significant increase in dehydrogenase and decrease in alkaline phosphomonoesterase activities after 24 h of treatment, which continued till 30 days. In seed treatments, imidacloprid and lindane were present in soil up to 90 and 120 days with average half-lives (t(1/2)) of 40.9 and 53.3 days, respectively. Within 90 days, imidacloprid residues were declined up to 73.17% to 82.49% while decline in lindane residues ranged from 78.19% to 79.86% within 120 days. In imidacloprid seed treated field, both dehydrogenase and phosphomonoesterase activities were increased between 15 and 60 days after sowing. However, a significant decreases in both dehydrogenase and phosphomonoesterase enzyme activities were observed between 15 and 90 days after lindane seed treatment.     

Cypermethrin induced histological changes in gonadotrophic cells, liver, gonads, plasma levels of estradiol-17beta and 11-ketotestosterone, and sperm motility in Heteropneustes fossilis (Bloch)

The aim of the present investigation is to assess the impact of cypermethrin on reproductive physiology in catfish, Heteropneustes fossilis during prespawning phase. Results indicate that there is a decrease in the size of gonadotrophic cells with less granulation, pycnosis in the liver, presence of immature oocytes and atretic follicles in the ovaries and gross condensation of spermatogenic cells in testes after cypermethrin exposure at sublethal concentration. The gonado-somatic index (GSI), plasma levels of estradiol-17beta (E2) and 11-ketotestosterone (11-KT) also decreases. The motility of sperm cells is dependent on the dilution (2000 times) and duration of motility is recorded 2min maximally at 90s after post-activation. The dose 0.1 and 0.01ppm is sublethal, while 1ppm is lethal on sperm motility. Results indicate that cypermethrin causes inhibition of reproduction by acting at the hypothalamo-hypophyseal-gonadal axis as is manifest from the histological observations of gonadotrophs along with disruption of follicular wall and spermatogenic cells. Obviously such changes are responsible for decreasing the steroid hormone levels which result in decreasing scale and duration of sperm motility after 45d exposure of cypermethrin in this species.     

Chronic effects of mercuric chloride on the activities of some enzymes in certain tissues of the fresh water murrel,

Alterations in the activities of some enzymes in the brain, gills, intestine, kidney, liver and muscles have been examined in the fresh water murrel, , after exposure to a sublethal concentration of mercuric chloride (3 μg/1) for 15, 30 and 60 days. The results revealed that after 15 days of exposure amino acid oxidase activity was elevated in brain and liver and inhibited in intestine. The activity of xanthine oxidase was increased in gills, and inhibited in kidney. Thirty days exposure produced significant inhibition in the activities of malate dehydrogenase in liver, glutamate dehydrogenase in gills and brain, aminoacid oxidase in gills, and xanthine oxidase in liver and intestine. In contrast, glutamate dehydrogenase in intestine, kidney and liver and aminoacid oxidase in brain and liver were elevated. After 60 days of treatment, a decrease in the activity of glucose-6-phosphatase was recorded in gills, intestine, kidney and liver. Hexokinase activity in kidney and liver, and malate dehydrogenase in all the six tissues were inhibited. Glutamate dehydrogenase activity in intestine, kidney and liver remained higher than in control fish. In brain, kidney and liver the activity of aminoacid oxidase was elevated, but in gills the enzyme activity decreased. Xanthine oxidase activity was inhibited in intestine and liver.     

Effect of commercial formulation of four organophosphorus insecticides on the ovaries of a freshwater teleost, Mystus vittatus (bloch)--a histological and histochemical study

The chronic effect of an exposure to a sublethal concentration of commercial formulation of 4 organophosphorus insecticides such as birlane (chlorfenvinphos), gardona (tetrachlorvinphos), phosdrin (mevinphos) and malathion on midvitellogenic ovaries of Mystus vittatus was investigated using several histological and histochemical techniques. The loss of stage II and III oocytes accompanied by a significant decline in gonadosomatic index was recorded. The vitellogenesis in treated fishes was ceased as the oocytes did not advance to further stages as evidenced by the absence of lipid yolks and the lack of a PAS positive follicular layer. The RNA content of remaining stage I oocytes of exposed fishes resembled that of the stage I oocytes of the control fishes. The absence of delta 5-3 beta-hydroxysteroid dehydrogenase suggested the lack of steroidogenesis. Reduced glucose-6-phosphate dehydrogenase activity was also observed. The significance of these observations has been discussed in light of available literature.     

Chronic effects of mercuric chloride on the activities of some enzymes in certain tissues of the fresh water murrel, Channapunctatus

Alterations in the activities of some enzymes in the brain, gills, intestine, kidney, liver and muscles have been examined in the fresh water murrel, $\text{Channa}$$\text{punctatus}$, after exposure to a sublethal concentration of mercuric chloride (3 μg/1) for 15, 30 and 60 days. The results revealed that after 15 days of exposure amino acid oxidase activity was elevated in brain and liver and inhibited in intestine. The activity of xanthine oxidase was increased in gills, and inhibited in kidney. Thirty days exposure produced significant inhibition in the activities of malate dehydrogenase in liver, glutamate dehydrogenase in gills and brain, aminoacid oxidase in gills, and xanthine oxidase in liver and intestine. In contrast, glutamate dehydrogenase in intestine, kidney and liver and aminoacid oxidase in brain and liver were elevated. After 60 days of treatment, a decrease in the activity of glucose-6-phosphatase was recorded in gills, intestine, kidney and liver. Hexokinase activity in kidney and liver, and malate dehydrogenase in all the six tissues were inhibited. Glutamate dehydrogenase activity in intestine, kidney and liver remained higher than in control fish. In brain, kidney and liver the activity of aminoacid oxidase was elevated, but in gills the enzyme activity decreased. Xanthine oxidase activity was inhibited in intestine and liver.     

Soil dehydrogenase, phosphomonoesterase and arginine deaminase activities in an insecticide treated groundnut (Arachis hypogaea L.) field

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2 pyridyl phosphorothioate) 20 EC and Quinalphos (O,O-diethyl O-quinoxalin-2-yl phosphorothioate) 25 EC, were applied in groundnut (Arachis hypogaea L.) field as seed treatment at 25 ml/kg and soil treatment at 4 l/ha in 1998 and 1999. The residues of these insecticides were monitored during the entire crop season and their effect on the soil enzymes dehydrogenase, phosphomonoesterase and arginine deaminase were studied. Ninety nine percent of chlorpyrifos residues were dissipated within 60 days from seed treated soil and 98% dissipation was observed in soil treated field for the same days. Its half lives in seed treated soil were 8 days and 7 days and in soil treated field were 9.2 days in and 7.5 days in 1998 and 1999 respectively. Dissipation of quinalphos in comparison to chlorpyrifos was slow both in seed treated and soil treated field. Eighty seven percentage to 92% dissipation of quinalphos residues were observed from seed treated soil and 98% residues were dissipated from soil treated field within 75 days. Its half lives in seed treated soil were 20 days and 18 days and in soil treated field, its half lives were 13 days and 17 days 1998 and 1999 respectively. Inhibition in dehydrogenase activity followed by recovery was observed both in seed and soil treatments with chlorpyrifos. An inhibition of 17.2% was estimated after 60 days of seed treatment in comparison to control. Dehydrogenase activity was significantly reduced to 63% after 15 days of quinalphos seed treatment in comparison to control in 1998. Similar trends were observed in 1999. A significant inhibition in dehydrogenase activity was observed after soil treatment both in 1998 and 1999. Phosphomonoesterase activities were significantly inhibited upto 25.2% as compared to the control, on the 15th day of chlorpyrifos seed treatment in 1998 and similarly, after one day of treatment in 1999. Quinalphos inhibited the phosphomonoesterase activity till the end of the experimental period in the soil treated fields, whereas recovered within 30-60 days of treatment in the seed treated fields. Arginine deaminase activity was significantly stimulated within one day after chlorpyrifos seed and soil treatments in both years. The activity was almost threefold higher on the 30th and the 15th day of soil treatment in 1998 and 1999, respectively. A temporary inhibition of arginine deaminase activity was observed after quinalphos treatment. It was observed that in most of cases insecticides have temporary inhibitory effect on soil enzymes. However, inhibition was smaller in seed treated soil than in direct soil treatment.     

Impairments of cadmium on vitellogenin accumulation in the hepatopancreas of freshwater crab <Emphasis Type="Italic">Sinopotamon henanense</Emphasis>

During ovary maturation of crabs, vitellogenin (Vg), a precursor molecule of vitellin (Vn) needed for embryogenesis, can be produced in large quantities in the hepatopancreas and then transported to the ovary by the hemolymph. In the present study, effects of Cd on Vg accumulation in the hepatopancreas and Vg transportation of the freshwater crab Sinopotamon henanense were investigated. We also studied the impacts of Cd on the mRNA expression of genes involved in energy metabolism, protein metabolism, and metallothionein (MT) and glutathione (GSH) synthesis. After Cd treatment, the Vg concentration and the Vg mRNA expression in the hepatopancreas were downregulated. Pearson’s correlation coefficient showed that the Vg level in the hepatopancreas correlated positively with those of the ovary and hemolymph (correlation coefficients 0.844 and 0.749, respectively), suggesting that the Vg transport from the hepatopancreas to the ovary can be impaired by Cd. The levels of carbohydrate and protein in the hepatopancreas of Cd-exposed crabs were decreased, and an inhibited protein metabolism was also observed. Energy production related isocitrate dehydrogenase and cytochrome C oxidase mRNA expressions, and MT and GSH synthesis increased after 10 days of Cd treatment and decreased after 20 days. Cd also caused a time-dependent upregulation of malondialdehyde. Our findings showed that Cd decreased Vg accumulation in the hepatopancreas due to partially excessive energy consumption and an activated defense system in the hepatopancreas, suggesting a possible regulatory mechanism in S. henanense which is the competitive advantage of energy reserves in metabolic Cd stress responses over the high-energy flux during vitellogenesis to ensure a continuous supply of metabolic energy. Moreover, the damage of Vg accumulation in the hepatopancreas caused by Cd could lead to an insufficient accumulation of Vn in the ovary and cause a retardation of oocyte development.     

Organochlorine pesticides in Avicennia marina from the Mumbai mangroves, India

This paper examines the concentration levels of organochlorine pesticides (OCPs) such as hexachlorocyclohexane (HCH) (α, β and γ), Aldrin, Endosulfan (α, β), DDE, DDD, and DDT in the leaf, root and sediment part of Avicennia marina, a dominantly distributed mangrove species in Mumbai. The overall pattern of accumulation of OCPs was observed as ∑HCH > ∑DDT > ∑Endosulfan > Aldrin. The occurrence of αHCH was found to be frequent in all the samples. The leaf and root samples showed better uptake of organochlorine pesticides whereas lower concentration levels of OCPs was observed in case of sediment samples. The mean concentration of OCPs was observed highest in case of root samples. This study indicated towards the capability of A. marina in bioaccumulation of the OCPs. The present data will serve as the baseline against which future study of OCPs uptake in the mangrove plant species may be assessed.     

Sublethal effects of diazinon on the structure of the testis of bluegill,Lepomis macrochirus: a microscopic analysis

A histopathological analysis of the testis exposed to diazinon, an organophosphorus pesticide was conducted. The testis from the control fish exhibited a normal histological structure with a round, oval or somewhat rectangular shaped with closely associated seminiferous tubules. The seminiferous tubules were surrounded by a layer of connective tissue with myoid cells, Leydig cells, blood vessels, lymph vessels and nerves. Upon exposure for 24 h, the overall structure of the testis looked disrupted and more often the sperm cells appeared to be somewhat larger, but spermatid were smaller than that of the control group. The 48 h exposure caused more dense and closely associated seminiferous tubules. The 72 h exposure induced dilation of the seminiferous tubules and their lumen. Ninety-six hours exposure induced more loose and disrupted structures. In this group (96 h), there was a significant reduction in both lumen and seminiferous tubule size in comparison with the control, 24, 48, and 72 h groups. After 2 weeks of exposure hardly any lumen was seen, which imparted more dense and compacted structures. The biometric analysis gave a highly significant increase in the diameter of the seminiferous lumen. After 48 h of exposure a significant reduction was seen in diameter in comparison with that of the control group. Seventy-two hour exposure indicated an increase in diameter, whereas 96 h exhibited another decrease. One week caused the diameter to increase whereas 2 weeks of exposure caused them to decrease again. The increase in diameter of the seminiferous tabule may be caused by weakening of the surrounding connective tissue and myoid cells, due to exposure to diazinon. The change in the diameters of the seminiferous tabules was very irregular. No apparent co-relation was found between the size of the fish, body weight and weight of the testis after they have been subjected to different exposure periods. There was significant change in the germ cells' diameters. As the germ cells are the essential first step in the process of spermatogenesis, their size reduction may hinder the production of viable spermatozoa, endangering the population dynamics. In order to find out the pathways of this pesticide effects, we propose to study the histopathology of pituitary gland a vital organ of the reproductive axis.     

Bioaccumulation studies of organochlorinated pesticides in tissues of Cyprinus carpio

Freshwater fish Cyprinus carpio was selected for the study of bioaccumulation of organochlorinated pesticides in tissues like gills, muscle, intestine, kidney, and liver in a continuous fed system. The pesticides used were Aldrin, Dieldrin, BHC, and DDT. The bioaccumulation of Dieldrin was maximum of 85.0 microg g(-1) wet weight in liver tissue while minimum of 7.30 microg g(-1) wet weight for DDT at 30 days exposure time. Bioconcentration factor (BCF) has followed the same trend in liver tissue for Dieldrin and DDT. The rate of bioaccumulation was found to be maximum of 4.3879 microg g(-1) wet weight in liver tissue and minimum of 0.0021 microg g(-1) wet weight in gill tissue for 30 days exposure. As evidenced by the increasing values of BCF, pesticide uptake also showed increased trend with the increase in exposure time. A high correlation coefficient ranging between 0.7247 and 0.9616 between the pesticide concentration and exposure time was observed. Based on actual BCF values, log Kow were calculated and the values are well within the reported values of 6.5 indicating efficient relationship between BCF and log Kow because beyond the 6.5 the bioconcentration levels off.     

Selenium for the mitigation of toxicity induced by lead in chicken testes through regulating mRNA expressions of HSPs and selenoproteins

Lead (Pb) is a toxic element and environmental pollutant. Pb toxicity and antagonistic effect of selenium (Se) on Pb have been deeply studied in mammals. The testis is one of the target organs of Pb in birds. The aim of this study was to investigate the mitigating effect of Se on Pb toxicity in chicken testes by determining messenger RNA (mRNA) expressions of 5 heat shock proteins (HSPs) and 25 selenoproteins. Sixty male chickens (7-day-old) were randomly divided into the control group, the Se group, the Pb group, and the Pb + Se group, and were fed for 90 days. The feeding methods of chickens were as follows: The control group was fed drinking water and commercial diet (0.49 mg/kg Se). Lead acetate was added into the drinking water (350 mg/L Pb). Sodium selenite was added into the commercial diet (1 mg/kg Se). Multivariate correlation analysis and principal component analysis (PCA) were used to define the relationships among all the measured factors and the most important parameters that could be used as key factors, respectively. The results indicated that Se decreased the increase of mRNA expressions of all the HSPs and increased the decrease of mRNA expressions of all the selenoproteins induced by Pb in the chicken testes. HSP70 may be a biomarker of Pb poisoning in the chicken testes. Se alleviated Pb-induced toxicity in the chicken testes through regulating mRNA expressions of HSPs and selenoproteins.

     

Oxidative stress and biochemical perturbations induced by insecticides mixture in rat testes

The joint action of pyrethroids, lambda-cyhalothrin (LC) in combination with organophosphates, fenitrothione (FNT) on antioxidant defense system and lipid peroxidation biomarkers in rat testes was studied. The results suggest that incubation of testes homogenate with different concentrations of insecticide mixture for different time intervals significantly decreased the activity of antioxidant enzymes, like glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT), and the level of reduced glutathione (GSH). In addition, a significant inhibition in transaminases (AST, ALT), phosphatases (AcP, AlP) activity and protein content were observed. On the other hand, FNT plus LC increased the cellular lipid peroxidation (LPO) level and the activity of lactate dehydrogenase (LDH). In conclusion, the use of insecticides mixture might cause marked oxidative damage in a concentration and time-dependent manner.     

Tissue bioaccumulation patterns, xenobiotic biotransformation and steroid hormone levels in Atlantic salmon (Salmo salar) fed a diet containing perfluoroactane sulfonic or perfluorooctane carboxylic acids

In the present study, groups of juvenile Atlantic salmon (Salmo salar) were fed gelatine capsules containing fish-food spiked with PFOA or PFOS (0.2 mg kg⁻¹ fish) and solvent (methanol). The capsules were given at days 0, 3 and 6. Blood, liver and whole kidney samples were collected prior to exposure (no solvent control), and at days 2, 5, 8 and 14 after exposure (Note: that day 14 after exposure is equal to 7 d recovery period). We report on the differences in the tissue bioaccumulation patterns of PFOS and PFOA, in addition to tissue and compound differences in modulation pattern of biotransformation enzyme genes. We observed that the level of PFOS and PFOA increased in the blood, liver and kidney during the exposure period. Different PFOS and PFOA bioaccumulation patterns were observed in the kidney and liver during exposure- and after the recovery periods. Particularly, after the recovery period, PFOA levels in the kidney and liver tissues were almost at the control level. On the contrary, PFOS maintained an increase with tissue-specific differences, showing a higher bioaccumulation potential (also in the blood), compared with PFOA. While PFOS and PFOA produced an apparent time-dependent increase in kidney CYP3A, CYP1A1 and GST expression, similar effects were only temporary in the liver, significantly increasing at sampling day 2. PFOA and PFOS exposure resulted in significant decreases in plasma estrone, testosterone and cortisol levels at sampling day 2, and their effects differed with 17α-methyltestostrerone showing significant decrease by PFOA (also for cholesterol) and increase by PFOS. PFOA significantly increased estrone and testosterone, and no effects were observed for cortisol, 17α-methyltestosterone and cholesterol at sampling day 5. Overall, the changes in plasma steroid hormone levels parallel changes in CYP3A mRNA levels. Given that there are no known studies that have demonstrated such tissue differences in bioaccumulation patterns with associated differences in toxicological responses in any fish species or lower vertebrate, the present findings provide some potential insights and basis for a better understanding of the possible mechanisms of PFCs toxicity that need to be studied in more detail.     

Induction of rat hepatic microsomal enzymes by toxaphene pretreatment

Abstract

The effects of pretreatment of rats with toxaphene on hepatic drug metabolizing enzymes and several other parameters of the mixed function oxidase system were investigated. Adult male Sprague‐Dawley rats were fed diets containing 0, 50, 100, 150 and 200 ppm of toxaphene for 14 days. The body weight gain was unaltered as well as the food consumption in all the toxaphene fed groups. There was no change in the weights of brain, kidney, heart, and testes but the liver weight was significantly increased. The thymus weight in all the toxaphene fed groups was decreased. Hydroxylation of pentobarbital and aniline was significantly enhanced in rats exposed to toxaphene. Ethylmorphine‐N‐demethy‐lase activity in the toxaphene treated rats was also elevated. Enhanced hydroxylation of pentobarbital was also evident from the decreased sleeping time following pentobarbital administration. Exposure to toxaphene increased cytochrome P‐450, NADPH‐cytochrome c‐reductase and dehydrogenase in hepatic microsomal fractions. The binding of aniline and hexobarbital to microsomes was also enhanced, suggesting that the intermediate steps in the electron‐transfer system were increased. In conclusion, pretreatment of rats with toxaphene for fourteen days resulted in the induction of the hepatic mixed function oxidase system.     

Estrogenic and antiestrogenic effect of in vitro treatment of follicular cells with 2,3,7,8-tetrachlorodibenzo-p-dioxin

OBJECTIVE: Two types of follicular cells from preovulatory ovary were cultured in vitro separately and in co-culture to test difference in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) action on particular cell types. METHODS: The accumulation of TCDD in follicular wall was analysed using coupled capillary gas chromatography mass spectrometry. Whole preovulatory follicles were isolated from ovary and incubated with prolonged exposure to 0.1 nM TCDD or single exposure to 10 nM TCDD for four days. In the second part of experiments direct effects of TCDD on steroidogenesis were investigated in porcine theca cells (Tc) and granulosa cells (Gc) cultured alone and in co-culture (GT). The media were collected after four days for steroid analysis. RESULTS: 59.3% and 81.2% of TCDD added to the culture medium was accumulated after 0.1 and 10 nM, respectively. TCDD in a dose-dependent manner increased estradiol secretion with concomitant progesterone secretion by theca interna cells. On the other hand decrease of both progesterone and estradiol secretion by granulosa cells cultured alone and in co-culture with theca cells was noted. CONCLUSION: Different cell-specific estrogenic or antiestrogenic effect of TCDD were found in ovarian follicles.