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.     

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.     

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.     

Lipid peroxidation and oxidative stress in rat erythrocytes induced by chlorpyrifos and the protective effect of zinc

Male and female rats were orally administered chlorpyrifos at a dose of 6.75 mg kg⁻¹ body weight for 28 consecutive days. An additional chlorpyrifos group received zinc (227 mg l⁻¹) in drinking water throughout the experimental duration. Two groups more served as controls; one received water only and the other received zinc in drinking water. Administration of chlorpyrifos resulted in a significant increase in lipid peroxidation (LPO) level and significant decrease in the activities of superoxide dismutase (SOD), glutathione-s-transferase (GST), catalase (CAT) and acetylcholinesterase (AChE) in erythrocytes of male and female rats. In contrast, zinc-chlorpyrifos treatment showed insignificant differences (p ? 0.05-0.01), compared to control results, regarding LPO, SOD, GST and CAT. In case of AChE, supplementation of zinc showed little alteration in the activity of this enzyme in the rats treated with chlorpyrifos. It can deduce that chlorpyrifos induced oxidative stress and lipid peroxidation in erythrocytes of male and female rats. The overall results reveal the pronounced ameliorating effect of zinc in chlorpyrifos-intoxicated rats and variation in the response of male and female animals regarding alteration in the level of some biochemical parameters and LPO.     

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.     

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

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

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.     

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).     

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.     

Functional expression of Helicoverpa armigera CYP9A12 and CYP9A14 in Saccharomyces cerevisiae

Elevated oxidative detoxification is a major mechanism responsible for pyrethroid resistance in Helicoverpa armigera from Asia. Constitutive overexpression of CYP9A12 and CYP9A14 was associated with pyrethroid resistance in the YGF strain of H. armigera. CYP9A12 and CYP9A14 were functionally expressed in the W(R) strain of yeast (Saccharomyces cerevisiae) transformed with a plasmid shuttle vector pYES2. The cell lysates prepared from yeast transformed with CYP9A12 and CYP9A14, respectively, exhibited considerable O-demethylation activities against two model substrates p-nitroanisole (0.59 and 0.42 nmol p-nitrophenol min⁻¹ mg protein⁻¹) and methoxyresorufin (2.98 and 5.41 pmol resorufin min⁻¹ mg protein⁻¹), and clearance activity against the pyrethroid esfenvalerate (8.18 and 4.29 pmol esfenvalerate min⁻¹ mg protein⁻¹). These results provide important evidence on the role of CYP9A12 and CYP9A14 in conferring pyrethroid resistance in H. armigera, and also demonstrate that the yeast expression system can provide necessary redox environment for insect P450s to metabolize xenobiotics.     

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%).     

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.     

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 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.     

Anti-oxidant response of Cucumis sativus L. to fungicide carbendazim

Our purpose in this research was to determine the response of anti-oxidative enzymes of cucumber (Cucumis sativus L.) when carbendazim applied as soil drench at 0, 5, 50, and 100 mg kg⁻¹. The changes in the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) enzymes were measured in roots, stems, and leaves of cucumber. The plants were sampling at cotyledon phase (14 days) and florescence phase (56 days). A strong correlation between anti-oxidant content and carbendazim concentration was observed. As concentration increased, the activities of SOD and CAT increased in roots and leaves. While in stems, SOD and CAT activities were increased in florescence phase and declined in cotyledon phase. The content of GPX increased in stems, and declined in leaves. Higher levels of SOD, CAT, and GPX were observed in cucumber cotyledons than the older leaves. The present study suggested that carbendazim treatments had different effects on cucumber anti-oxidant system in different tissues. It was concluded that cotyledons might play an important role for adaptation as the carbendazim concentration increased, and the ability of mature cucumber to maintain a balance between the formation and detoxification of activated oxygen species appeared likely to enhance. On the basis of our observations, we conclude that increased SOD, CAT and GPX activity provides plant with increased carbendazim stress tolerance.     

Soil applied propiconazole alleviates the impact of salinity on Catharanthus roseus by improving antioxidant status

An investigation was carried out to estimate the salt stress ameliorative effect of propiconazole (a triazole group of fungicide cum plant growth regulator) on the NaCl stressed Catharanthus roseus. The experiment was conducted in pots. All the pots were irrigated to the field capacity with ground water up to 30 days after sowing (DAS). The treatments were given as 80 mM NaCl, 80 mM NaCl + 20 mg l⁻¹ propiconazole and 20 mg l⁻¹ propiconazole alone on 30, 50 and 70 DAS. The samples were collected on 60 and 80 DAS. NaCl stress inhibited the root and stem length, total leaf area, fresh and dry weight, and antioxidant enzymes superoxide dismutase (SOD), peroxidase (POX) and polyphenol oxidase (PPO) activities. Plants treated with NaCl with propiconazole increased these parameters to a larger extent when compared to NaCl stressed plants. The propiconazole treatment increased the root length, fresh and dry weight, and SOD, POX and PPO activities, but decreased the stem length and leaf area when compared to control. The results showed that the propiconazole could significantly ameliorate the effect of NaCl stress in Catharanthus roseus.     

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.     

Inhibitory effects of validamycin compounds on the termites trehalase

Trehalase, with the target to control insects, nematodes and fungi, is of increasing interest and has been investigated extensively in recent years. Validamycin compounds, as competitive trehalase inhibitors and lead compounds with broad applications have attracted substantial attention as well. In this study, the characterizations of termites trehalase were investigated and the inhibitory effects of validamycin compounds on the termites trehalase were studied as well. Results showed that the termites trehalase is presumably belonging to the acid trehalase with optimal pH of 3.3 and optimal temperature of 37 °C. It was investigated that the concentrations of validoxylamine A (VAA), validoxylamine B (VBB), validamycin A (VA) and validamycin B (VB) required for 50% inhibition IC₅₀ of termites trehalase were calculated to be 14.73 mg l⁻¹, 20.80 mg l⁻¹, 3.17 × 10³ mg l⁻¹and 2.24 × 10³ mg l⁻¹, respectively. The inhibition kinetic constant K_i values for the above validamycin compounds were 3.2 × 10⁻⁶ mol l⁻¹, 1.03 × 10⁻⁵ mol l⁻¹_, 4.02 × 10⁻⁴ mol l⁻¹and 2.69 × 10⁻⁴ mol l⁻¹, respectively. Validoxylamine A appeared to be the most potential termites trehalase inhibitor among the four compounds.     

Impact of malathion stress on lipid metabolism in Limnonectus limnocharis

Despite mounting concerns about amphibian population declines, information on impact of pesticides on physiological changes is meager. The present study deals the influence of an organophosphate pesticide—malathion on the lipid metabolism of Limnonectus limnocharis under laboratory conditions. Changes in the lipid metabolism were analyzed in the liver, muscle, ovary, and testis of frogs exposed to lethal (10.67 mg L⁻¹ for 1, 2, 3, and 4 days) and sub-lethal (2.13 mg L⁻¹ for 1, 5, 10, 15, 20, and 25 days) concentrations of malathion. Upon lethal concentration treatment, against the increase of fatty acids, glycerol, and lipase activities in all tested tissues, there was decrease in the total lipids content over different durations. On the other hand, exposure to sub-lethal concentration, the amount of total lipids content, free fatty acids, glycerol and lipase activity increased. Changes in the lipid metabolism due to lethal concentration of malathion exposure could depict the negative impact on the reproductive success, which would result in decline of amphibian population.