Suwarna Pandey

Oxidative stress biomarkers of exposure to deltamethrin in freshwater fish, Channa punctatus Bloch

The pyrethroid class of insecticides, including deltamethrin, are being used as substitutes for organochlorines and organophosphates in pest-control programs because of their low environmental persistence and toxicity. Ecotoxicological consequences of deltamethrin, particularly its effects on antioxidants in fish and other aquatic organisms, are not well understood. We investigated the effect of deltamethrin (0.75 microg/L) on antioxidants in a freshwater fish, Channa punctatus Bloch, using standard laboratory conditions. A single exposure for 48 h caused induction of various antioxidant enzymes and nonenzymatic antioxidants in kidney and liver. The induction of these antioxidants was not very prominent in gills. In fact, certain antioxidants were found to be depleted in gills. Catalase activity was decreased in all the tissues. Deltamethrin also induced lipid peroxidation in all the tissues, gills showing the highest levels. Glutathione, which is an established nonenzymatic antioxidant in fish, was significantly (P<0.001) increased in all the tissues. Ascorbic acid content increased in kidney and liver while it decreased in gills. The findings of the present investigation show that deltamethrin has oxidative-stress-inducing potential in fish, and gills are the most sensitive organs. It is also interesting to note that gills are the primary sites of deltamethrin absorption and their antioxidant potential is also very poor. The various parameters studied in this investigation can also be used as biomarkers of exposure to deltamethrin. It is suggested that appropriate ecotoxicological risk assessment should be made in the areas where deltamethrin is proposed to be used in pest control activities.

Biomarkers of oxidative stress: a comparative study of river Yamuna fish Wallago attu (Bl. & Schn.)

Various oxidative stress biomarkers in gill, kidney and liver tissues in the Indian freshwater fish Wallago attu (Bl. & Schn.) were investigated. Fish were collected from two sites along the river Yamuna, which differ in their extent and type of pollution load. A comparison was made between the biomarker responses and general water chemistry at the two sites. The oxidative stress biomarkers that were analyzed included superoxide dismutase (SOD), catalase (CAT), xanthine oxidase (XOD) and glutathione redox cycle enzymes viz., glutathione peroxidase (GPx), glutathione reductase (GR) and glucose 6-phosphate dehydrogenase (G6PD). Levels of reduced glutathione (GSH) and lipid peroxidation (LPO) were also evaluated. All biomarkers; SOD (P<0.001 in liver, kidney and gill), XOD (P<0.01 in kidney and P<0.001 in liver and gill), GR (P<0.01 in liver, P>0.05 in kidney and P<0.001 in gill), G6PD (P<0.001 in liver, P>0.05 in kidney and P<0.01 in gill), GSH (P<0.001 in liver, kidney and gill) and LPO (P>0.05 in liver, kidney and gill) were found to be substantially higher in the fish collected from Panipat when compared with values in tissues of fish collected from Agra site. GPx and CAT showed a varied response. GPx activity was higher (P<0.001) in gills and kidney of the fish collected at Panipat site. However, liver showed significant low values (P<0.01) when compared with Agra site values. CAT activity was found to be significantly low, in both liver (P<0.01) and kidney (P<0.001) whereas in gills non-significant (P>0.05) low values were observed. Water chemistry data at two sites indicated that Panipat site with higher biochemical oxygen demand, chemical oxygen demand, pH and low dissolved oxygen was comparatively more polluted than Agra site. Industrial activity profile of both the sites also indicates that Panipat has vigorous industrial activity coupled with intensive use of chemicals in agricultural practices in Haryana state. The findings of the present investigation provide a rational use of oxidative stress biomarkers in aquatic ecosystem pollution biomonitoring. This is also the first such attempt reported from India.

Effects of exposure to multiple trace metals on biochemical, histological and ultrastructural features of gills of a freshwater fish, Channa punctata Bloch

The trace metals are frequently encountered as mixtures of essential and non-essential elements. Therefore, evaluation of their toxic effects individually does not offer a realistic estimate of their impact on biological processes. We studied effects of a mixture of four essential and toxic metals (Cu, Cd, Fe and Ni) on biochemical and morphological characteristics of the gills of a biomarker freshwater fish Channa punctata (Bloch) using environmentally relevant concentrations. Fish were exposed to metal mixture through tank water for 7, 15 and 30 days. Biochemical studies as well as light microscopy (LM) and scanning electron microscopy (SEM) revealed significant metal exposure-induced alterations in gills. Besides ultastructural changes, activities of antioxidant enzymes such catalase (CAT), glutathione S-transferase (GST) and superoxide dismutase (SOD) were significantly altered in the gills of exposed fish. The reduced glutathione (GSH) was significantly (p<0.001) decreased, while lipid peroxidation (LPO) was significantly (p<0.001) increased. The main alterations in general morphology of fish gills included spiking and fusion of secondary lamellae, formation of club-shaped filaments, and vacuolization and necrosis of filament epithelium in the interlamellar regions. SEM studies showed gradual increase of the density and apical surface area of the chloride cells and transformation of the surface structure of the pavement cells. The results of this study indicate adaptive as well a toxic responses in fish gills exposed to mixture of trace metals. Low concentrations of trace metal appear to compromise the antioxidant defense of gills. Lesions in the gill morphology caused by the effect of low concentrations of trace metals could lead to functional alterations and interference with fundamental processes such as maintenance of osmoregulation, gas exchange and xenobiotic metabolism in the exposed fish populations.

IMMUNOMODULATORY EFFECTS OF FENUGREEK (TRIGONELLA FOENUM GRAECUM L.) EXTRACT IN MICE

Immunomodulatory activity of aqueous extract of Trigonella foenum graecum L., a widely used medicinal and dietary herb, was evaluated in male Swiss albino mice. Mice were treated with three doses of extract (50, 100 and 250 mg/kg body weight per os) for 10 days. Body weight, relative organ weight, cellularity of lymphoid organs, delayed type of hypersensitivity (DTH) response, plaque-forming cell (PFC) assay, haemagglutination titre (HT), quantitative haemolysis of SRBC (QHS) assay, phagocytosis, and lymphoproliferation were studied in various groups of animals. At doses of 50 and 100 mg/kg, a significant increase (p < 0.05) in relative organ weight of thymus was observed but there was no effect on kidney and spleen weights. Liver weight also increased significantly at doses of 100 and 250 mg/kg. However, no elevation in the levels of liver function test (LFT) enzymes was observed. As regards lymphoid organ cellularity, spleen recorded no significant increase at any dose, whereas cellularities of thymus and bone marrow were significantly increased. T. foenum graecum extract elicited a significant (p < 0.001) increase in the DTH response at doses of 50 and 100 mg/kg, but the change at higher dose of 250 mg/kg was not statistically significant. Humoral immunity as measured by PFC showed an elevated response at a dose of 100 mg/kg, but at 50 and 250 mg/kg, no significant effect was observed. In the HT test, plant extract also showed modulatory effect at all the doses. Plant extract elicited a significant increase in phagocytic index and phagocytic capacity of macrophages. Stimulatory response of plant extract was also observed in lymphoproliferation assay but the response was weak. Overall, T. foenum graecum showed a stimulatory effect on immune functions in mice. As it is used for a variety of medicinal purposes, its immunostimulatory effect, as reported in this study, strengthens the rationale of its use in several Ayurvedic and Unani drugs.

Aqueous extract of walnut (Juglans regia L.) protects mice against cyclophosphamide-induced biochemical toxicity.

Walnut (Juglans regia L.) is extensively used in traditional systems of medicine for treatment of various ailments. It is described as an anticancer, tonic, blood purifier and detoxifier agent. The present study was undertaken to investigate modulatory effects of walnut extract on the toxicity of an anticancer drug, cyclophosphamide (CP) with special reference to protection against disruption of drug metabolizing and antioxidant enzymes. Plant extract+CP group animals showed restoration in the level of cytochrome P450 (CYP) content and in the activities of glutathione S-transferase (GST), glutathione peroxidase (GP) and catalase (CAT) in both liver and kidneys. But plant extract restored the activity of super oxide dismutase (SOD) and the level of reduced glutathione (GSH) in the kidneys only when compared with CP-treated animals. Plant extract treatment alone caused significant reduction in the content of CYP in the kidneys mainly. The extract showed a significant increase in the level of GSH and in the activities of GP in both the tissues and CAT in liver only, whereas no significant change was observed in the activities of GST and SOD. CP treatment resulted in a significant (P<0.01) increase in the lipid peroxidation (LPO) in the liver and kidneys compared with controls, while the extract CP treated group showed a significant decrease in the LPO in liver and kidneys when compared with the CP-treated group. The study shows that the use of J. regia extract might be helpful in abrogation of CP toxicity during the chemotherapy.

Protective effects of Emblica officinalis Gaertn. in cyclophosphamide-treated mice.

Cyclophosphamide (CP) is one of the most popular alkylating anticancer drugs in spite of its toxic side effects including immunotoxicity, hematotoxicity, mutagenicity and a host of others. The present study was undertaken to assess the protective effects of total aqueous extract of a medicinal plant, Indian gooseberry (Emblica officinalis Gaertn.) in mice treated with CP. These protective effects were studied on immunological parameters and kidney and liver antioxidants. Plant extract treatment at a dose of 100 mg/kg body weight per os (p.o.) for 10 days resulted in the modulation of these parameters in normal as well as CP (50 mg/kg)-treated animals. Plant extract in particular was very effective in reducing CP-induced suppression of humoral immunity. Plant extract treatment in normal animals modulated certain antioxidants of kidney and liver. In CP-exposed animals, plant pretreatment provided protection to antioxidants of kidney. Not only were the reduced glutathione levels significantly (p<0.001) increased but plant extract treatment resulted in restoration of antioxidant enzymes in CP-treated animals. It is suggested that E. officinalis or its medicinal preparations may prove to be useful as a component of combination therapy in cancer patients under CP treatment regimen. Human & Experimental Toxicology (2001) 20, 643–650.

Modulatory effect of copper on nonenzymatic antioxidants in freshwater fish Channa punctatus (Bloch.).

Effect of the low level of copper exposure on nonenzymatic antioxidants was studied in a freshwater fish Channa punctatus (Bloch.). Fish were exposed to cupric chloride at the concentration of 10 ppb for 4 wk (28 d) in a static culture condition. Copper significantly (p < 0.001) increased the serum ceruloplasmin level and total iron-binding capacity. A significant (p < 0.05) increase in reduced glutathione level was recorded in all of the tissues. With regard to nonprotein thiols, copper decreased their level in the liver, but increased it in the gill. The protein-bound thiols remained unaltered except for an increase in the liver. Metallothionein (MT) induction was observed in liver only. Copper exposure had no significant effect on the ascorbic acid level and induced no lipid peroxidation over control values. It is suggested that by modulating the ceruloplasmin level, copper indirectly protects the fish, as it facilitates conversion of pro-oxidant iron to nonoxidant iron. It also induces an array of antioxidants that may be beneficial to fish in the case of oxidative stress resulting from chemical pollutants.