Goat serum as an alternative to establish cell culture from Indian major carp, Cirrhinus mrigala

Serum from goat, calf, and chicken sources were evaluated in terms of attachment, growth, and proliferation of explants of Indian major carp, Cirrhinus mrigala. The attachment of explants viz. heart, liver, and kidney was directly proportional to the concentration of the serum. Among these sera, the highest percentage of attachment, growth, and proliferation was recorded for 10% goat serum and 15% newborn calf serum without affecting their cell morphology. On contrary to these sera, chicken serum at 15% concentration was found to be mildly toxic for all the explants. The cell count was significantly high for the kidney, liver, and heart at 10% goat serum among all the tested sera as well as concentration. Similarly, the liver, heart, and kidney explants were found to survive up to the tenth, seventh, and ninth passage, respectively. Therefore, the goat serum at 10% concentration can be used as effectively as newborn calf serum for routine culture of fish cells.     

Responses of isolated scale melanophores of a fresh water carp,Cirrhinus mrigala (Ham.) to zidovudine

Zidovudine (AZT) induced concentration related aggregation in C. mrigala melanophores. Denervated melanophores failed to respond to AZT. Specific and nonspecific alpha adrenoceptor antagonists completely blocked the responses of fish melanophores to AZT. Histamine and prostaglandin antagonists also inhibited aggregation of the melanophores induced by AZT. The results suggest that AZT may release a mixture of neurotransmitter like substances, which cause the aggregation of this fish melanophores.     

On the origin, development and probable functions of taste bud in the lip and bucco-pharyngeal epithelia of an Indian freshwater major carp, Cirrhinus mrigala (Hamilton) in relation to food and feeding habits.

1. Various developmental stages of taste bud have been observed in the fry, fingerling and adult of Cirrhinus mrigala (Hamilton). 2. The fries are zooplankton-feeders (carnivorous) and sight-feeders. Therefore, functional taste buds are absent in the lip and bucco-pharyngeal epithelia. Only formative stages of taste bud are present. Some epithelial cells aggregate together to form this structure which has no function in gustation. Moreover, the fires have low R. L. G. valle (0.71;..1.20) due to the carnivorous diet. 3. In the fingerling, the stages of elongation, differentiation and maturation of taste bund develop from the formative stage present in the fries in order to adjust to changed food and feeding habit. The value of R. L. G; (1.21...3.10) also increases. 4. Fully formed taste buds along with the stage of differentiation are found in the adult stage. The adult fish becomes herbivorous and bottom-feeder. Consequently, the value of R.L.G. 3.11...12) becomes highest in this stage.     

Seasonal changes in the blood parameters of two major carps, Labeo rohita (Ham.) and Cirrhina mrigala (Ham.).

Seasonal changes in blood properties of two Indian major carps, L. rohita and C. mrigala, were investigated. Haematocrit values, haemoglobin concentrations and erythrocyte numbers showed seasonal fluctuations, being high from September to November and again in April-May, and low from January to March. Males had generally higher blood values than the females.     

Identification of microsatellite DNA markers for population structure analysis in Indian major carp, Cirrhinus mrigala (Hamilton-Buchanan, 1882)

Forty‐four primer sequences available for four cyprinid fishes were tested to amplify microsatellite loci in Indian major carp, Cirrhinus mrigala. A total of 12 loci were successfully amplified with clear scorable patterns and five thereof were polymorphic. Suitability of the identified polymorphic loci in population structure analysis of C. mrigala was assessed. Genetic variation was examined in 76 specimens collected from five different rivers. The mean observed heterozygosity ranged from 0.247 to 0.333. Significant heterogeneity in allele frequencies was detected, indicating that the samples analysed did not belong to homogenous populations. The identified microsatellite markers are promising for the analysis of intraspecific divergence in C. mrigala across its distribution range.     

A histochemical study on the adrenal components of the teleost Cirrhinus mrigala (Ham.)

The adrenal components of C. mrigala are embedded in the pronephric cephalic kidney around the post cardinal vein. The cortical cells responded positively to the lipids, ascorbic acid, delta 5-3 beta-HSD, G-6-PD, MAO, acid and alkaline phosphatase tests. The presence of intense MAO activity may suggest the possible involvement of monoamines in the adrenocortical function. Localization of lipids and delta 5-3 beta-HSD show the sites of corticosteroid synthesis. In the chromaffin cells, MAO, acid and alkaline phosphatase activity was moderate whereas they gave a strong reaction to ascorbic acid test in comparison to the cortical cells. Noradrenaline (NA) and adrenaline (A) storing cells were differentiated adopting glutaraldehyde silver, dichromate and iodate techniques. NA and A storing cells are almost totally depleted of their contents after reserpine treatment. The histochemical response of the adrenal gland of this species is largely comparable to that of higher vertebrates.     

Dietary requirement of tryptophan for growth and survival of the Indian major carp, Cirrhinus mrigala (Hamilton-Buchanan) fry

The experimental diets (40% protein) contained graded levels of tryptophan (0.60, 0.75, 1.00, 1.25, 1.50, 1.75%) of dietary protein. Each test diet was fed to triplicate groups of mrigal fry twice daily at 10% of the total biomass for 8 weeks. The dietary tryptophan requirement was estimated by plotting weight gain against dietary levels of tryptophan using two separate regression equations, the point of intersection of two equations was taken as optimum level, which occurred at 0.48% of the diet (1.20% of dietary protein).     

Short-term mercury exposure on Na+/K+-ATPase activity and ionoregulation in gill and brain of an Indian major carp,Cirrhinus mrigala

Recently mercury pollution has been increased considerably in aquatic resources throughout the world and it is a growing global concern. In this study, the 96 h LC50 value of waterborne mercuric chloride for Cirrhinus mrigala was found to be 0.34 mg/L (with 95% confidence limits). Fingerlings of C. mrigala were exposed to 0.068 and 0.034 mg/L of mercuric chloride for 96 h to assess the Na⁺/K⁺-ATPase activity and ionoregulation (Na⁺, K⁺ and Cl^?) in gill and brain. Results showed that Na⁺/K⁺-ATPase activity and ionic levels (Na⁺, K⁺ and Cl^?) in gill and brain of fish exposed to different concentrations of mercuric chloride were found to be significantly (p < 0.05) decreased throughout the study period. Mercury inactivates many enzymes by attaching to sulfur atoms in which the enzyme Na⁺/K⁺-ATPase is highly sensitive to mercury. The inhibition of gill and brain Na⁺/K⁺-ATPase activity might have resulted from the physicochemical alteration of the membrane due to mercury toxicity. Moreover, inhibition of Na⁺/K⁺-ATPase may affect the ion transport and osmoregulatory function by blocking the transport of substances across the membrane by active transport. The present study indicates that the alterations in these parameters can be used in environmental biomonitoring of mercury contamination in aquatic ecosystem.     

Bioactive forms of gonadotropin releasing hormone in the brain of an Indian major carp,Catla catla (Ham.)

Two forms of biologically active gonadotropin releasing hormones were isolated from the hypothalami ofCatla catla. Gonadotropin releasing hormone activity was studiedin vitro using enzymatically dispersed carp pituitary cell incubation system. Gonadotropin released into the medium was measured by carp gonadotropin-radio immuno assay. Acetic acid extracted hypothalamic material was subjected to acetone fractionation. Among the three protein pellets obtained at different time periods (ACI, ACII and ACIII), AC II exhibited the gonadotropin releasing hormone activity. Gel filtration of AC II through Sephadex G-25 column showed three protein peaks (SG I, SG II SGIII) and only S G II demonstrated strong gonadotropin releasing hormone activity. Elution of SG II through FPLC Mono Q column (an anion exchanger) in NaCl gradient programme showed one unadsorbed (MQ I) and three adsorbed (MQ II, MQ III and MQ IV) protein peaks. MQ III, which was eluted with 51% NaCl, exhibited gonadotropin releasing hormone activity. Surprisingly, unadsorbed fractions, MQ I, also showed gonadotropin releasing hormone activity. MQ 1 was therefore subjected to FPLC Mono S (a cation exchanger) column chromatography where a highly active gonadotropin releasing hormone enriched peak, i.e., MS III, could be eluted with 45% NaCl. These findings show thatCatla catla hypothalamus has two forms of gonadotropin releasing hormones one anionic (carp gonadotropin releasing hormone I) and another cationic (carp gonadotropin releasing hormone II). These two forms of gonadotropin releasing hormones were also active in heterologous carp species, rohu(Labeo rohita), mrigal(Cirrhinus mrigala) and an exotic common carp(Cyprinus carpio). Combined activity of two forms of gonadotropin releasing hormones was significantly greater as compared to any of the single form.     

The immunosuppressive effect of alpha-permethrin on Indian major carp, rohu (Labeo rohita Ham.)

The immunosuppressive effects of bath exposure to a sub lethal concentration of the synthetic pyrethroid alpha-permethrin (3.05 x 10(-4) mg l(-1)) in the Indian Major carp, Labeo rohita was studied after 45 days' exposure. In some groups, the effects of alpha-permethrin on non-specific defences and serum enzymes of carp were investigated after challenge with Aeromonas hydrophila. Several nonspecific immune responses and serum enzymes were reduced after exposure of alpha-permethrin. Bactericidal activity of rohu serum was reduced significantly in pesticide and bacteria treated fish. The Glutamic Oxaloacetate Transaminase (GOT) and Glutamic Pyruvate Transaminase (GPT) activity were increased in immunosuppressed fish. Blood glucose level was elevated significantly and Hb% was reduced significantly in pesticide and bacteria treated fishes as compared to the negative control.     

Evidence for presence of GABA-ergic receptor mediated dispersion in isolated scale melanophores of a carp,Cirrhinus mrigala Ham

Effects of GABA-ergic agonists and antagonists were examined on the melanophores of a carp C. mrigala in vitro. GABA and baclofen both induced concentration - related dispersion in fish melanophores. Denervation of the melanophores by reserpine treatment potentiated the sensitivity of the melanophores to GABA. While denervation by cooling treatment inhibited the sensitivity of the melanophores to GABA, atropine, bicuculline and pentylenetetrazole all inhibited the dispersal responses of the melanophores induced by higher concentrations of GABA. 5-aminovaleric acid also significantly inhibited the dispersion of the melanophores induced either by GABA or baclofen. It is concluded that GABA-ergic agonist induced dispersal responses in C mrigala melanophores are mediated through specific GABA receptors. The presence of both GABAA and GABAB receptors in this fish melanophores has been indicated.     

Isolation and characterization of immunoglobulin of the Indian major carp, rohu [Labeo rohita (Ham.)

Information on the structure and character of immunoglobulin of fishes is essential in health management. A study was carried out to characterize the serum immunoglobulin (IgM) of the Indian major carp, rohu Labeo rohita (Ham.). Rohu (500g) were immunised with bovine serum albumin (BSA) and the anti-BSA antibody was purified employing BSA-CL agarose affinity column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified Ig in a 3% gel under non-reduced conditions revealed a single protein having a molecular weight of 850kDa. Analysis of the purified serum in 10% SDS-PAGE under reduced conditions revealed that the immunoglobulin contained heavy and light chains with molecular weights of 85 and 23kDa, respectively. A polyclonal mouse anti-rohu IgM was prepared and used in an immunodot test which showed a specific reaction of the crude rohu anti-BSA antiserum and the purified anti-BSA IgM with BSA. Results indicate that the immunoglobulin of L. rohita is tetrameric IgM, similar to that of other fishes.     

Seasonal patterns of feeding of juvenile major carp, Labeo rohita (Ham.) in a Bangladesh pond

The results of the studies on the seasonal patterns of feeding of 203 Labeo rohita (Ham.) taken by cast and seine net for one year from a Bangladesh pond are presented. Studies were based on the frequency of occurrence and the points method. A change in the volume of food with change of seasons was noted. The amounts of food eaten were greater for the warmer than for other months and very low during winter indicating an upturn in late spring with a higher level continuing during the summer.
Organic detritus formed the bulk of the gut contents throughout the study period. On a seasonal basis, phytoplankton and aquatic macrophytes were the group of organisms frequently ingested but the former was more important. Animal food was scarce but appreciable quantities of sand and mud were found.     

Phosphorus requirements and optimum calcium/phosphorus ratio in the diet of mrigal Cirrhinus mrigala (Ham.) fingerlings

An experiment was conducted to investigate phosphorus requirements and the optimal calcium/phosphorus (Ca/P) ratio on growth and carcass tissue composition of mrigal Cirrhinus mrigala (c. 6 g). Five purified diets were formulated to contain Ca/P ratios of 1 : 0 (0.35 : 0), 1 : 1 (0.35 : 0.35), 1 : 2 (0.31 : 0.63), 1 : 3 (0.24 : 0.71), and 1 : 4 (0.19 : 0.75), respectively. Growth performance and feed conversion ratio of mrigal responded significantly (P < 0.01) to the Ca =0.19 : P = 0.75 diet. Carcass protein, lipid, and P percentage also increased significantly with the higher P level. Based on this study, it may be concluded under the given conditions that the optimum Ca : P ratio in mrigal feed is 0.19 : 0.75.     

Annual cyclical changes in the testicular activity of an Indian freshwater major carp, Labeo rohita (Hamilton)

Different male germ cells identified on the basis of histological and cytological characteristics in the testicular lobules of Labeo rohita have been grouped into spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids and spermatozoa. The seasonal changes of the testis in L. rohita have been described according to its morphological peculiarities as well as to its variations in gonadal volumes, GSI values and frequency percentages of the different male germ cells occurring in the testicular lobules. Consequently, the entire testicular cycle in L. rohita may be categorised into 4 distinct phases viz., growth, maturation of pre-spawning, spawning, and resting or post-spawning phases.     

Acute and sublethal effects in an Indian major carp Cirrhinus mrigala exposed to silver nitrate: Gill Na+/K+-ATPase, plasma electrolytes and biochemical alterations

Due to prolonged use of silver in many applications, it enters into the freshwater and affects the aquatic organisms. Fingerlings of Cirrhinus mrigala were exposed to acute and sublethal concentrations of silver nitrate and the alterations of gill Na(+)/K(+)-ATPase, plasma electrolytes and biochemical parameters were assessed. The median lethal concentration of silver nitrate to the fish C. mrigala for 96 h was found to be 0.107 mg/l (with 95% confidence limits). 1/10th of LC 50 96 h value (0.0107 mg/l) was selected for sublethal study. During acute treatment branchial Na(+)/K(+)-ATPase activity was inhibited approximately 44.34% after 96 h of exposure. In sublethal treatment, silver nitrate could not produce a significant change in the activity of the enzyme at the end of 7th day. However, after 14th day, significant (p < 0.05) decrease was noted showing 22.52%-49.11% in rest of the study period. Silver intoxication resulted hyponatremia, hypokalemia, hypochloremia, and hypoproteinemia in both the treatments. Despite the decrease in these parameters, plasma glucose level was found to be increased in both the treatments to endure the silver toxicity. We suggest that the alterations in branchial Na(+)/K(+)-ATPase activity, plasma electrolytes, and biochemical parameters of fish may be useful in environmental biomonitoring and to assess the health of fish in freshwater habitat contaminated with silver.