Toxic effects of acephate on Paramecium caudatum with special emphasis on morphology, behaviour, and generation time

The continuous increase in the number of new chemicals as well as the discharges of solid and liquid wastes triggered the need for simple and inexpensive bioassays for routine testing. In recent years, there has been increasing development of methods (particularly rapid tests) for testing environmental samples. This paper describes the quick toxic evaluation of an organophosphorus insecticide, acephate (O,S-dimethyl acetylphosphoramidothioate) on Paramecium caudatum for acute and sub-acute toxicity studies with reference to morphology, behaviour, and its generation time. The lethal concentrations for 10 min and 2 h were determined by probit method, as 500 mg L⁻¹ and 300 mg L⁻¹, respectively. Higher concentrations of 10 min exposure caused cell lysis with disintegration of cell membrane and precipitation of protoplasm. Combination of conventional light microscopy and computerized video tracking systems were used to study the locomotor behaviour of paramecia. The test organism was under stress and exhibited an initial increase and subsequent decrease in the swimming speed when exposed to 1/4, 1/2, 3/4, and LC₅₀ concentrations for 10 min (125, 250, 375, and 500 mg L⁻¹, respectively). Similar changes were also noticed when paramecia were exposed to LC₅₀ for 2 h. In a separate set of experiments, the number of generations and generation time in 24 h was evaluated with respect to the different sub-lethal concentrations (30, 60, 120, and 240 mg L⁻¹). The number of generations decreased and generation time extended significantly in a concentration dependent manner. The results indicate that the Paramecium toxicity assay could be used as a complimentary system to rapidly elucidate the cytotoxic potential of insecticides. The major advantages associated with these tests are: they are inexpensive, simple, user-friendly, space saving, and seem to be attractive alternatives to conventional bioassays.     

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.     

Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus

The present study was conducted to determine the 96 h-LC₅₀ of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L⁻¹) was administered for 96 h to determine the LC₅₀. The 96 h-LC₅₀ value of benomyl was 4.39 (3.23-5.60) mg L⁻¹ in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L⁻¹) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL⁻¹) level, MCV (μm³), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL⁻¹) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL⁻¹) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min⁻¹ mg protein⁻¹), and SOD (U mg protein⁻¹) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L⁻¹, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.     

Metalaxyl-induced changes in the antioxidant metabolism of Solanum nigrum L. suspension cells

The antioxidant responses of Solanum nigrum L. cell suspension cultures to metalaxyl exposure were investigated. An increase in lipid peroxidation and hydrogen peroxide content, for both concentrations tested (20 mg L⁻¹; 40 mg L⁻¹) revealed the response of oxidative metabolism of cell suspensions to metalaxyl. Superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) activities increased, particularly in the highest concentration of metalaxyl used. An analysis by non-denaturing polyacrylamide gel (PAGE) followed by staining for enzyme activity, revealed seven SOD isoenzymes, two CAT isoenzymes, and nine APX isoenzymes. Metalaxyl levels were quantified in the culture medium and results suggest that suspension cells were able to accumulate and/or degrade the fungicide five hours after exposure. SOD, CAT and APX isoenzymes were differently affected by the metalaxyl treatment. Results suggest that the higher concentration of metalaxyl induced oxidative stress to cell suspension cultures of S. nigrum.     

Effect of terbutryn at environmental concentrations on early life stages of common carp (Cyprinus carpio L.)

The aim of this study was to assess the toxicity of terbutryn to early developmental stages of common carp (Cyprinus carpio). Based on accumulated mortality in the experimental groups, lethal concentrations of terbutryn were estimated at 36 day LC50 = 3.06 mg l⁻¹ terbutryn. Based on inhibition of growth in the experimental groups, lowest observed-effect concentration (LOEC) = 0.005 mg l⁻¹ terbutryn; and no observed-effect concentration (NOEC) = 0.0007 mg l⁻¹ terbutryn. Fulton’s condition factors were significantly lower in fish exposed to 2 mg l⁻¹ compared with controls. By day 30, fish exposed to 0.00002 mg l⁻¹ - real environmental concentration in Czech rivers - 0.02, 0.2, and 2 mg l⁻¹ terbutryn showed significantly lower mass and total length compared with controls. No significant negative effects on hatching or embryo viability were demonstrated at the concentrations tested, but significant differences in early ontogeny among groups were noted. Fish from the two highest tested concentrations (0.2 and 2 mg l⁻¹) showed a dose-related delay in development compared with the controls. At concentrations of 0.02, 0.2, and 2 mg l⁻¹ damage to caudal kidney tubules when compared to control fish was found.     

Metabolic and histological parameters of silver catfish (Rhamdia quelen) exposed to commercial formulation of 2,4-dichlorophenoxiacetic acid (2,4-D) herbicide

The objective of this study was to investigate the effects of commercial formulation of herbicide 2,4-D on metabolic parameters, acetylcholinesterase (AChE) activity and liver histological evaluation of silver catfish (Rhamdia quelen) exposed for 96 h. AChE activity increased in brain (600 and 700 mg L⁻¹) and decreased in all concentrations tested in muscle tissue. Hepatic glycogen was reduced after 2,4-D exposure ranging from 47.67% (400 mg L⁻¹) until 59.3% (700 mg L⁻¹). Hepatic tissue showed lactate reduction at all 2,4-D concentrations tested and glucose was reduced only at 700 mg L⁻¹. In the highest concentration tested hepatic glycogen and glucose reduced instead plasma glucose levels increased. White muscle tissue showed glycogen reduction in fingerlings exposed to all herbicide concentrations and glucose reduction at 700 mg L⁻¹. Muscle lactate levels increase at all 2,4-D concentrations tested. Vacuolation of hepatocytes and changes in its arrangement cords were observed by histologic analysis in group treated with 700 mg/L of 2,4-D. These results suggest that silver catfish exposed to concentrations of 2,4-D near of CL₅₀ showed metabolic and histological response to compensate some stress caused by herbicide exposure. Taken together parameters measured can be used as biomarkers to monitor herbicide contaminated water.     

Oxidative stress and locomotor behaviour response as biomarkers for assessing recovery status of mosquito fish, Gambusia affinis after lethal effect of an organophosphate pesticide, monocrotophos

Recovery study was performed at regular intervals to establish the time course of 50% and 100% recovery in neurotransmitter enzyme (acetylcholinesterase, AChE, EC 3.1.1.7) and locomotor behaviour response of mosquito fish, Gambusia affinis exposed to lethal concentration (20.49 mg L⁻¹) of an organophosphorous pesticide, monocrotophos (MCP) for 96 h. In vitro AChE activity studies indicated that MCP could cause 50% inhibition (I₅₀) at 10.2 × 10⁻⁵ M. A positive correlation was observed between brain AChE activity and swimming speed during the recovery study. Also, the recovery response of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) as well as lipid peroxidation (LPO) as biomarkers of oxidative stress were assessed in viscera of G. affinis. The results showed that the MCP besides its inhibitory effect on target enzyme AChE activity and induction in antioxidant enzyme activities as a characteristic of oxidative stress, which can be used as biomarkers in the pesticide contaminated aquatic streams.     

Individual and combined effects of dimecron-ziram on the levels of serum prolactin and selected minerals of an edible freshwater fish, Oreochromis mossambicus (Peters)

The individual and combined effects of dimecron (an organophosphate pesticide) and ziram (a carbamate fungicide) on the levels of serum prolactin and selected minerals (Ca, P, and Mg) are studied in an edible freshwater fish, Oreochromis mossambicus (Peters) exposed to sub-lethal concentrations of individual dimecron (0.002 mL L⁻¹) and ziram (0.008 mL L⁻¹) and combined 1:4 mixture of dimecron-ziram (0.01 mL L⁻¹) for 24, 48, and 72 h. Exposure of fish to sub-lethal concentration of individual and mixture of dimecron and ziram caused changes in the levels of serum prolactin and minerals studied compared to control fishes. Based on the results obtained, it was concluded that the individual exposures of dimecron and ziram comparatively manifested higher toxic effects (than the combined exposure) by reducing prolactin levels and loss of mineral contents. These changes may result in a severe osmoregulatory problem.     

Effects of the organophosphate pesticide Folidol 600® on the freshwater fish, Nile Tilapia (Oreochromis niloticus)

Nile Tilapia (Oreochromis niloticus) juveniles were exposed to different concentrations of Folidol 600® in static toxicity tests. The 24, 48, 72 and 96 h LC₅₀ values of Folidol 600® to O. niloticus were 17.82, 8.91, 4.00 and 2.70 mg L⁻¹, respectively. The values of hematological parameters increased, and inhibition of cholinesterases activity (AChE, BChE and PChE) in plasma of fish exposed to the higher concentrations of pesticide reached 94%. Furthermore, the exposure of Tilapia to Folidol 600® caused an increase of 4%, 20% and 38.4% in oxygen consumption at 0.1, 0.5 and 1.0 mg L⁻¹, respectively. However, exposure to 2.5, 5.0 and 10 mg L⁻¹ caused a decrease of 33.6%, 35.2% and 42.4% in oxygen consumption relative to the control. The ammonium excretion of fish exposed to 0.0, 0.1, 0.5, 1.0, 2.5, 5.0 and 10.0 mg Folidol 600®/L was 0.12, 0.18, 0.30, 0.33, 0.37, 0.36 and 0.33 μg/g/min, i.e., 50%, 150%, 175%, 208%, 200% and 175% increase, respectively, relative to the control.     

Acute toxicity of organophosphorous pesticide diazinon and its effects on behavior and some hematological parameters of fingerling European catfish (Silurus glanis L.)

Diazinon is commonly used for pest control in the agricultural fields surrounding freshwater reservoirs. So this study was conducted to determine the acute toxicity of this organophosphorous pesticide, contaminating aquatic ecosystems as a pollutant, and its effects on behavior, and some hematological parameters of fingerling European catfish, Silurus glanis. Diazinon was applied at concentrations of 1, 2, 4, 8, 16, 32, and 64 mg L⁻¹. The water temperature in the experimental units was kept at 16 ± 1 °C. The number of dead fishes significantly increased in response to diazinon concentrations 2-64 mg L⁻¹ (p < 0.05). With increasing diazinon concentrations, the fishes exposed duration 1 to 96 h significantly increased the number of dead fishes (p < 0.05 for each cases). The 1, 24, 48, 72, and 96 h LC₅₀ values (with 95% confidence limits) of diazinon for fingerling European catfish were estimated as 14.597 (12.985-16.340), 12.487 (11.079-14.471), 8.932 (7.907-10.348), 6.326 (no data because of p > 0.05), and 4.142 (no data because of p > 0.05) mg L⁻¹, respectively. Compared to the control specimens, fish after an acute exposure to diazinon was significantly lower erythrocyte, leukocyte, hemoglobin, hematocrit, MCV, MCH, and MCHC values (p < 0.05). In addition, it was also showed a significantly negative correlation between these hematological parameters and exposure times of diazinon (p < 0.01).     

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.     

Design, synthesis and antifungal/insecticidal evaluation of novel nicotinamide derivatives

A series of nicotinamide derivatives based on Boscalid were prepared and tested for their activities against seven plant pathogenic fungi and two insects. The preliminary bioassays indicated that almost all of the synthesized target compounds displayed the antifungal activities and some of them also had certain insecticidal activities. And, compound 12 showed the strongest activity of all against Rhizoctonia solani (EC₅₀ = 0.010 mg L⁻¹) and Sphaceloma ampelimum (EC₅₀ = 0.040 mg L⁻¹), even stronger than Boscalid, a new nicotinamide fungicide. Additionally, both compounds 1 and 2 showed strong activities against Plutella xylostella (90% and 80%, respectively, at 1000 mg L⁻¹).     

Effect of chitosan on growth and plasma membrane properties of Rhizopus stolonifer (Ehrenb.:Fr.) Vuill

The effect of chitosan (0.5, 1.0 and 2.0 mg ml⁻¹) on mycelium growth, potassium efflux, pH of the incubation medium, and activity of plasma membrane H⁺-ATPase of Rhizopus stolonifer was evaluated. The mycelium growth of R. stolonifer was reduced on media amended with chitosan at 1.0 and 2.0 mg ml⁻¹. The highest antifungal index (65%) was obtained on media containing chitosan at 2.0 mg ml⁻¹. Potassium efflux induced by the addition of the three chitosan concentrations was demonstrated. The highest potassium effluxes were observed with addition of chitosan at 2.0 mg ml⁻¹. R. stolonifer grown 72 h on Potato Dextrose Broth without chitosan generated a decrease in the medium’s pH from 5.4 to 4.8. However, the addition of chitosan at all concentrations caused increases on pH of medium cultures. The highest pH increases were observed on media amended chitosan at 2.0 mg ml⁻¹ resulting in the raising of pH level to 6.9. Additionally, there was an important inhibitory effect of chitosan (1.0 and 2.0 mg ml⁻¹) on H⁺-ATPase activity in the plasma membrane of R. stolonifer (32.70%).     

Induction of defense response of Oryza sativa L. against Pyricularia grisea (Cooke) Sacc. by treating seeds with chitosan and hydrolyzed chitosan

Seeds of rice (Oryza sativa L.) were treated with chitosan and hydrolyzed chitosan at 100, 500 and 1000 mg L⁻¹. After 18 days of germination, spore suspension of Pyricularia grisea was applied. The enzyme activity of phenylalanine ammonia-lyase, β-1-3-glucanase, chitinase and chitosanase in leaves of rice seedlings was evaluated after 24, 72, 120 and 168 h of inoculation. Blast affected area (%) was evaluated 7 and 14 days after spraying spore suspension. Chitosan performance to elicit defense response induction was associated with the concentration and type of chitosan. The activity of most of the enzymes tested was induced in leaves of treated seeds with chitosan and hydrolyzed chitosan at 1000 and 500 mg L⁻¹, respectively. The highest enzyme activities were observed with hydrolyzed chitosan after 72 h however, compared to chitosan, the activity was not maintained during the entire post-inoculation period. The highest control (0 = no lesions) of P. grisea in rice seedlings was observed at 1000 mg L⁻¹ in both chitosan and hydrolyzed chitosan treated leaves. Symptoms of infection by P. grisea were evident after 14 days evaluation date, but according to the standard scale proposed by the International Rice Research Institute, these symptoms fell into the resistance category of blast diseases.     

Inhibitory effects of Schiff analogs of salicylidene aniline on phenoloxidase from Pieris rapae L. (Lepidoptera: Pieridae)

In order to gain insight into the development of insecticides with novel modes of action, the effects of salicylidene aniline (a), salicylidene-4-chloroaniline (b), salicylidene-4-bromoaniline (c), and salicylidene-4-nitroaniline (d) on partially purified phenoloxidase (PO) from Pieris rapae L. were investigated. The results showed that the 4 compounds could inhibit PO activity, and the inhibitor concentrations leading to a loss of 50% activity (IC₅₀) were estimated to be 0.025 mmol L⁻¹, 0.732 mmol L⁻¹, 0.471 mmol L⁻¹, and 0.675 mmol L⁻¹, respectively. Meanwhile, all the inhibitors showed reversible competitive inhibition, except (d), which showed reversible mixed inhibition. The K_I values were determined as 0.106 mmol L⁻¹, 10.059 mmol L⁻¹, 8.390 mmol L⁻¹, and 20.198 mmol L⁻¹ for the four compounds, respectively. The UV-vis spectra of (a) and (d) in the presence of copper ions and the enzyme showed that (a) could directly chelate the copper ions of PO; however, (d) could neither chelate the additional copper ions nor the copper ions of PO.     

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.     

Acute toxicity of diazinon on the common carp (Cyprinus carpio L.) embryos and larvae

In the present study, the toxic effects on the embryos and larvae of the common carp were used as a model to investigate the organophosphorus pesticide, diazinon, which contaminates aquatic ecosystems. Data obtained from the diazinon acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The control and six test experiments were repeated five times. The number of dead embryos significantly increased in response to diazinon concentrations 0.25, 0.5, 1, 2, 4, and 8 mg L⁻¹ (p < 0.05 for each case). The 48 h LC₅₀ value (with 95% confidence limits) of diazinon for common carp embryos was estimated at 0.999 (0.698-1.427) mg L⁻¹. Dose-response decreases in hatching success were recorded as 84.60, 75.2, 54.1, 31.0, 6.0, and 0.0%, respectively (p < 0.05). The number of dead larvae significantly increased with increasing diazinon concentrations exposed for 24-96 h (p < 0.05). The 24, 48, 72, and 96 h LC₅₀ values (with 95% confidence limits) of diazinon for common carp larvae were estimated at 3.688 (2.464-8.495), 2.903 (2.019-5.433), 2.358 (1.672-4.005), and 1.530 (1.009-3.948) mg L⁻¹, respectively. There were significant differences in the LC₅₀ values obtained at different exposure times (p < 0.05). The results of the study suggest that low levels (0.25 mg L⁻¹) of diazinon in the aquatic environment may have a significant effect on the reproduction and development of carp.     

Oxidative parameters of Rhamdia quelen in response to commercial herbicide containing clomazone and recovery pattern

In this study, fish Rhamdia quelen, were exposed to different concentrations of herbicide clomazone: 0.0 (control), 0.45 and 0.91 mg L⁻¹. After exposure for 8 days to herbicide, fish were transferred to clean water for a recovery period (8 days). Oxidative stress indicators such as thiobarbituric acid reactive substances (TBARS) levels and protein carbonyl content, as well as antioxidant defenses, such as catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), ascorbic acid and non-protein thiols levels were studied, using the liver, brain and muscle tissues. Herbicide exposure increased TBARS in muscle and in liver at higher concentration. In liver protein carbonylation increased and catalase activity did not change in fish exposed to herbicide. SOD enhanced in liver at concentration of 0.91 mg L⁻¹. GST, ascorbic acid and non-protein thiols levels increase at both concentrations. At the end of the recovery period the most of the parameters recovered whereas GST and ascorbic acid remain elevated. The present study demonstrates the occurrence of disorders in antioxidant parameters and importance in the assessment of the potential risk of herbicides as clomazone on fish species.     

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.     

Cytogenetic effects of commercially formulated atrazine on the somatic cells of Allium cepa and Vicia faba

In the present study cytogenetic effects of atrazine herbicide, were examined on the root meristem cells of Allium cepa and Vicia faba. Test concentrations were chosen by calculating EC₅₀ values of formulated atrazine against both the test systems which determined to be 30 mg l⁻¹ for A. cepa and 35 mg l⁻¹ for V. faba, respectively. For cytogenetic effects root meristem cells of A. cepa were exposed to 15, 30 or 60 mg l⁻¹ whereas V. faba to 17.5, 35 or 70 mg l⁻¹ for 4 or 24 h. Roots exposed for 4 or 24 h, after sampling, were left in water for 24 h recovery and sampled at 24 h post-exposure. A set of onion bulbs or seedlings of V. faba exposed to DMSO (0.3%) was run parallel for negative control. Treatment of atrazine significantly and dose-dependently inhibited the mitotic index (MI) and induced micronucleus formation (MN) chromosome aberrations (CA) and mitotic aberrations (MA) in both the test systems at 4 or 24 h. Root meristem cells examined at 24 h post-exposure also revealed significant (p < 0.001) frequencies of MN, CA or MA despite considerable decline. Chromosome breaks and fragments were found to be major CA whereas C-metaphase, chromosome bridges and laggards were prevalent MA. Results of our study, indicate that atrazine may produce genotoxic effects in plants. Further, both the plant bioassays found to be sensitive indicators for the genotoxicity assessment as the outcome of majority of in vivo/in vitro mammalian tests are comparable.