Effect of arsenic (AsIII) on glutathione-dependent enzymes in liver and kidney of the freshwater fish Channa punctatus

The possible role of glutathione-dependent enzymes in the liver and kidney of the freshwater fish Channa punctatus has been studied after exposure to arsenic trioxide for different durations. Activities of glutathione-S-transferases, glutathione peroxidase, glutathione reductase, and catalase decreased in the liver and kidney as a result of the initial increase in arsenic concentration in the liver and kidney. However, during longer exposures, a decline in arsenic concentration corresponded with improved enzyme activity. Because arsenic manifests its toxicity by inducing oxidative stress, the antioxidant enzymes, especially the glutathione-dependent enzymes, play a protective role in arsenic toxicity.     

Cell volume changes affect gluconeogenesis in the perfused liver of the catfish<Emphasis Type="Italic">Clarias batrachus</Emphasis>

In addition to lactate and pyruvate, some amino acids were found to serve as potential gluconeogenic substrates in the perfused liver ofClarias batrachus. Glutamate was found to be the most effective substrate, followed by lactate, pyruvate, serine, ornithine, proline, glutamine, glycine, and aspartate. Four gluconeogenic enzymes, namely phosphoenolpyruvate carboxykinase (PEPCK), pyruvate carboxylase (PC), fructose 1,6-bisphosphatase (FBPase) and glucose 6-phosphatase (G6Pase) could be detected mainly in liver and kidney, suggesting that the latter are the two major organs responsible for gluconeogenic activity in this fish. Hypo-osmotically induced cell swelling caused a significant decrease of gluconeogenic efflux accompanied with significant decrease of activities of PEPCK, FBPase and G6Pase enzymes in the perfused liver. Opposing effects were seen in response to hyperosmotically induced cell shrinkage. These changes were partly blocked in the presence of cycloheximide, suggesting that the aniso-osmotic regulations of gluconeogenesis possibly occurs through an inverse regulation of enzyme proteins and/or a regulatory protein synthesis in this catfish. In conclusion, gluconeogenesis appears to play a vital role inC. batrachus in maintaining glucose homeostasis, which is influenced by cell volume changes possibly for proper energy supply under osmotic stress.     

Studies on intestinal protease: Isolation,purification and effect of dietary proteins on alkaline protease activity of the air-breathing fish,Clarias batrachus (Linn.)

1. The effect of dietary protein levels on the proteolytic activity in the intestines of the air-breathing fish, Clarias batrachus (Linn.) has been studied
2. Activity of proteolytic enzymes increased significantly in fishes maintained with a 50% protein diet from those maintained with a 25% protein diet; still higher dietary protein percentage showed no further stimulation of enzyme activity.
3. In a study on the determination of sub-cellular localisation, it has been found that protease activity is more prominent in lysosomes than in other organelles of the cell.
4. A sixty fold purification of alkaline protease from the intestine of Clarias batrachus has been achieved by ion exchange chromatography on DEAE cellulose which has been further checked by polyacrylamide gel electrophoresis.

     

Resistance to oxidative stress in a freshwater fish Channa punctatus after exposure to inorganic arsenic

The biochemical toxicity of arsenic trioxide (As^III) in a freshwater edible fish Channa punctatus has been studied on exposures ranging from 7 to 90 d. The arsenic concentration increased exponentially in liver, kidney, gills, and muscles of fish up to 60 d of exposure to arsenic. However, arsenic concentration in these tissues declined at 90 d of exposure. This relationship between period of exposure and concentration of arsenic in selected tissues suggests an adaptive response of fish to arsenic. Furthermore, exposure to arsenic-induced lipid peroxidation in these organs increased initially at 7 d of exposure; however, it decreased up to 60 d of exposure but increased again at 90 d of treatment. Values of reduced glutathione (GSH) reflected the observations of lipid peroxidation. The role of GSH in this adaptive response has been discussed.     

Monoclonal antibody to serum immunoglobulins of Clarias batrachus and its application in immunoassays

Serum immunoglobulins of Clarias batrachus (Cb-Ig) were purified by affinity chromatography using bovine serum albumin as capture ligand. Under reducing conditions in SDS-PAGE, Cb-Ig was composed of a heavy (H) chain (68.7 kDa) and two light (L) chains (27.4 and 26.3 kDa). Purified Cb-Ig was used to produce a monoclonal antibody (MAb) designated E4 MAb that belonged to IgG1 subclass. In Western blotting, this MAb showed binding to H chain of purified Cb-Ig and putative H chains in reduced sera of C. batrachus, Clarias gariepinus and Heteropneustes fossilis. However, no binding was observed with serum protein of Labeo rohita and Channa striata. Cross-reactivity of anti-Cb-Ig MAb was observed with serum of C. batrachus, C. gariepinus and H. fossilis in competitive ELISA. In immunoblotting of non-reduced Cb-Ig with E4 MAb, four bands assumed to be tetrameric, trimeric, dimeric and monomeric form were observed. In flow cytometric analysis of the gated lymphocytes, the number of surface Ig-positive (Ig +) cells in blood, spleen, kidney and thymus of C. batrachus was determined to be 50.1 ± 3.1, 55.1 ± 3.36, 42.4 ± 4.81 and 5.1 ± 0.89%, respectively, using E4 MAb. Ig + cells were also demonstrated in formalin-fixed paraffin embedded tissue sections of spleen, kidney, thymus and smears of blood mononuclear cells in indirect immunoperoxidase test. The developed MAb was employed to detect pathogen-specific immunoglobulins in the sera of C. batrachus immunized with killed Edwardsiella tarda, by an indirect ELISA. This monoclonal antibody can be useful tool in immunological research and assays.     

Effect of stocking density on growth and survival of Clarias batrachus (Linn.) larvae and fry during hatchery rearing

Experiments were conducted to determine optimum stocking density for Clarias batrachus larvae and fry during hatchery rearing. The increase in stocking density decreased the total weight, specific growth rate (SGR) and percent weight gain of Clarias larvae during a 13‐day experiment. Survival rate was highest at a stocking density of 1000 m^?2 and lowest at 5000 m^?2. Stocking density did not influence the total biomass production of larvae. Clarias batrachus fry performance was studied during a 28‐day hatchery rearing experiment whereby fry stocked at a density of 100 m^?2 attained the highest total body weight (P < 0.05). The survival rate greatly declined to 59–61% by a density increase to 300 m^?2 and above. Stocking density influenced growth and survival of C. batrachus larvae and fry during hatchery rearing. The best performance was obtained when larvae were stocked at 2000 m^?2; survival was highest with C. batrachus fry stocked at 200 m^?2.     

Vanadium induced changes in glycogen content of Clarias batrachus (Linn) and Channa punctatus (Bloch)

Clarias batrachus (Linn) and Channa punctatus (Bloch) were exposed to 5, 10 and 15 ppm of vanadium and investigated the effects on tissue glycogen at 3, 6 and 9 hrs. Results indicated the variations in the level of glycogen were highest at 15 ppm after 9 hr exposure in liver than muscle, brain, kidney and heart of Clarias batrachus as compared to Channa punctatus. The variation recorded in tissue glycogen content was discussed in relation to respiratory distress, formation of mucus on the whole fish and disturbed behaviour of the fish.     

Studies in pteridophytes XVIII. Use of Salvinia auriculata aublet-an obnoxious weed-in paper industry

Clarias batrachus, an air breathing fish, when exposed to the mercuric chloride, shows enhanced activity of alkaline phosphatase in its dendritic organs. However, an inhibiting effect has been observed on the activity of acid phosphatase.     

Effect of Pinealectomy on Re-entrainment of Air-gulping Activity in Indian Walking Catfish,Clarias batrachus

The present study was aimed to examine the effect of pinealectomy on re-entrainment of air-gulping activity in Indian walking catfish Clarias batrachus under various LD phases. Live Clarias batrachus of mixed sex (40-50g body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into 2 groups. The pineal was surgically removed from one group (P_xG) and the other group was treated as intact control group (IG). Both groups were exposed under LD 12:12 cycle inside the chronocubicles. In every 7 days LD schedule was delayed by 4 hours by lengthening of the light-on time. Both intact and pinealectomized animals are exhibited 24h rhythm in air-gulping behaviour and its entrainability irrespective of phase shifts of LD cycle. However, the phase angle of peak with reference to light on hour gradually decreased from phase 1 to phase 4, irrespective of treatment (intact/pinealectomized). Further, it has been noticed that the occurrence of multifrequency rhythm (24h, 12h and 6h) is being decreased from phase 1 to phase 4. It could be concluded that daily light-dark cycle plays an important role in modulating the rhythmic characteristics of air-breathing activity in Clarias batrachus and that pinealectomy does not modulate the effects of LD cycles on such activity.     

Pinealectomy Does Not Modulate the Characteristics of 24-h Variation in Air-Gulping Activity of Clarias batrachus

The effect of pinealectomy on the characteristics of circadian variation in air-gulping activity of Clarias batrachus was examined. Live Clarias batrachus of mixed sex (40–50 gm body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into two groups. The pineal was surgically removed from fishes in the first group (P×G) and the second group was left intact (IG). The individuals were kept in the specially designed glass aquaria. They were exposed under LD 12:12 (Lights on at 06:00) inside the chronocubicles for recording of the air-gulping activity. Results clearly indicate that the air-gulping activity exhibits a significant 24 h rhythm in both Intact (IG) and Pinealectomized (P×G) groups of this catfish. The peak of the air-gulping activity rhythm was located in the dark phase of the LD cycle in both groups. It may be concluded that pinealectomy does not modulate the 24 h rhythm in air-gulping activity of Clarias batrachus under LD 12:12.     

Origin and Development of Neutrophils in an Air Breathing Fish,Channa punctatus Bloch

Mahajan, C. L., Dheer, J. M. S. 1980. Origin and development of neutrophils in an air breathing fish, Channa punctatus Bloch. (Department of Zoology, University of Rajasthan, India.) — Acta zool. (Stockh.) 61(4): 221–224. An account of the study of the development of neutrophils based on detailed examinations of kidney and spleen imprints has been given in this paper. Each developmental stage has been investigated with respect to its morphology, morphometry and cytochemistry using certain special techniques developed in this laboratory for fishes such as differential haemoglobin staining, autoradiography etc. Five main stages (large lymphoid haemoblasts, promyelocytes, mesomyelocytes, metamyelocytes, mature neutrophils) have been identified in development and are described in detail.     

Do thyroid and testis modulate the effects of pineal and melatonin on haemopoietic variables inClarias batrachus?

Involvement of pineal and its major hormone, melatonin, in the process of erythropoiesis in a freshwater catfish,Clarias batrachus has been investigated. The study was conducted during four phases, namely preparatory phase, spawning phase, postspawning phase and late postspawning phase of its annual reproductive cycle. During each phase a fish received either melatonin injections or subjected to pinealectomy. In addition, each fish in all the groups, received either iopanoic acid or cyproterone acetate or vehicle in the morning or late afternoon. Results clearly indicate that melatonin stimulates the rate of erythropoiesis inClarias batrachus. It appears that the extent of stimulation depends upon the phase of the annual reproductive cycle. However, in general, the pineal-or melatonin-induced modulation of blood variables is gonad dependent and thyroid seems to play a time of the day dependent subtle role     

Development of EST derived SSRs and SNPs as a genomic resource in Indian catfish, <Emphasis Type="Italic">Clarias batrachus</Emphasis>

Clarias batrachus, an Indian catfish species, is endemic to the Indian subcontinent and potential cultivable species. The genomic resources in C. batrachus in the form of ESTs containing microsatellite repeats (EST-SSR) and single nucleotide polymorphisms (SNPs) that are associated with the expressed genes from spleen were mined. From a total of 1,937 ESTs generated, 1,698 unique sequences were obtained, out of which 221 EST-SSRs were identified and 54% could be functionally annotated by similarity searches. A total of 23 contigs containing 3 or more ESTs were found to contain 31 SNP loci, out of which 8 ESTs showed similarity to genes of known function and 1 for hypothetical protein. Nine ESTs with SSRs and/or SNPs identified in this study were reported to be associated with diseases in human and animals. These identified loci can be developed into markers in C. batrachus, which can be useful in linkage mapping, comparative genomics studies and for its genetic improvement programmes.     

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrin-induced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (∼12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), non-protein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues.     

Retinal Pigment Epithelium–Calycal Processes Association in the Catfish Clarias batrachus(Linnaeus, 1758)

Abstract In the duplex retina of the catfish Clarias batrachus(Linnaeus, 1758), the apical processes of the pigment epithelial cells have been found by transmission electron microscopy to be in intimate contact with the calycal processes around the basal portion of the photoreceptor outer segments. It is hypothesized that the retinal pigment epithelium effectively transports synthesized products and metabolites to the photoreceptor inner segments via the anatomical zone of the apical–calycal processes interface in this species.     

Seasonal reorganization of nitrogen metabolism in an Indian air-breathing teleosts,Channa punctatus (Bloch)

Channa punctatus, an air-breathing freshwater teleost, mobilizes more protein for its energy requirement during summer and spawning months, as revealed by the data on endogenous nitrogen excretion in the form of ammonia-N, urea-N, free amino acids, creatinine and creatine.     

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 June 2012–31 July 2012

This article documents the addition of 96 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Clarias batrachus, Marmota himalayana, Schizothorax richardsonii, Sitophilus zeamais and Syagrus romanzoffiana. These loci were cross‐tested on the following species: Clarias dussumeri, Clarias gariepinus, Heteropneustus fossilis, Sitophilus granarius and Sitophilus oryzae.     

Physiological responses to acute experimental hypoxia in the air-breathing Indian catfish, Clarias batrachus (Linnaeus, 1758)

With an aim to study the mechanism of adaptation to acute hypoxic periods by hypoxia-tolerant catfish, Clarias batrachus, the mass-specific metabolic rate (VO₂) along with its hematological parameters, metabolic response and antioxidant enzyme activities were studied. During progressive hypoxia, C. batrachus was found to be an oxyconformer and showed a steady decline in its aquatic oxygen consumption rate. When C. batrachus was exposed for different periods at experimental hypoxia level (0.98?±?0.1 mg/L, DO), hemoglobin and hematocrit concentrations were increased, along with decrease in mean cellular hemoglobin concentration, which reflected a physiological adaptation to enhance oxygen transport capacity. Significant increase in serum glucose and lactate concentration as well as lactate dehydrogenase activity was observed. Antioxidant enzymes were found to operate independently of one another, while total glutathione concentration was unaffected in any of the tissues across treatments. These observations suggested that hypoxia resulted in the development of oxidative stress and C. batrachus was able to respond through increase in the oxygen carrying capacity, metabolic depression and efficient antioxidant defense system to survive periods of acute hypoxia.     

Studies on the enzymatic activities related to varied pattern of diets in the air-breathing cat fish,clarias batrachus (Linn.)

1. Specific activity of amylase, cellulase, protease and lipase in the intestines of the air-breathing catfish, Clarias batrachus (Linn.) has been studied.
2. Excepting amylase and protease, the activity of lipase and cellulase showed practically no changes with change in the nutritional status of the diets.
3. pH optima of all enzymes were between 6.9 and 7.6
4. There is reason to believe from cellulase and high amylase activity in the intestine of the species that its culture operation could be done more economically by giving them a supplementary diet from indigeneously available raw material particularly from plant origin.