Dietary histidine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week growth trial was conducted to determine the dietary histidine requirement of the Indian major carp, Cirrhinus mrigala fingerling (length 4.22 ± 0.45 cm; weight 0.61 ± 0.08 g; n = 40). Isonitrogenous (400 g kg^?1 crude protein) and isoenergetic (17.90 kJ g^?1 gross energy) diets with graded levels of l ‐histidine (2.5, 5.0, 7.5, 10.0, 12.5 and 15.0 g kg^?1 dry diet) were formulated using casein and gelatin as a source of intact protein, supplemented with l ‐crystalline amino acids. Twenty fish were randomly stocked in 70‐L indoor polyvinyl circular fish tank (water volume 55‐L, water exchange rate 1–1.5 L min^?1) and fed experimental diets at the rate of 5% of their body weight/day divided over two feedings at 08:00 and 16:00 h. Maximum live weight gain (295%), best feed conversion ratio (FCR) (1.48) and protein efficiency ratio (PER) (1.69) occurred at 7.5 g kg^?1 of dietary histidine level. When live weight gain, FCR and PER data were analysed using second‐degree polynomial regression, the break points indicated histidine requirements at 9.4, 8.6 and 8.5 g kg^?1 of dry diet respectively. Significantly (P < 0.05) higher whole body protein and low moisture values were recorded at 7.5 g kg^?1 histidine level. Body fat increased significantly (P < 0.05) with increasing histidine levels. However, at 7.5 and 10 g kg^?1 histidine diets body fat did not differ (P > 0.05) to each other. Ash content of fish fed diets containing various levels of histidine did not differ except at 2.5 and 5.0 g kg^?1 inclusion levels where significantly (P < 0.05) higher ash was recorded. Protein deposition was also found to be significantly (P < 0.05) higher in the 7.5 g kg^?1 histidine diet. Based on the polynomial regression analysis of FCR and PER data, it is recommended that the diet for fingerling C. mrigala should contain histidine at 8.5 g kg^?1 of dry diet, corresponding to 21.25 g kg^?1 of dietary protein for optimum growth and efficient utilization of feed.     

Dietary threonine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Indian major carp fingerling, Cirrhinus mrigala (3.85±0.75 cm, 0.52±0.21 g), were fed isonitrogenous and isocaloric diets (40% crude protein, 4.28 kcal g^?1, gross energy) containing casein, gelatin and crystalline amino acids with graded levels of l ‐threonine (1.00, 1.25, 1.50, 1.75, 2.00 and 2.25 g 100 g^?1, dry diet) to determine the dietary threonine requirement. The feeding trial was conducted in triplicate for 8 weeks. Diets were fed twice a day at 08:00 and 16:00 hours at 5% body weight day^?1. The ration size and feeding schedule were worked out before the start of the feeding trial. Highest weight gain (304%) and best feed conversion ratio (1.43) were evident in fish fed diet containing 1.75% dietary threonine. Second‐degree polynomial regression analysis of weight gain, feed conversion ratio and protein efficiency ratio data indicated the dietary threonine requirement to be at 1.84%, 1.81% and 1.78%, respectively, corresponding to 4.60%, 4.52% and 4.45% of dietary protein. Minimum carcass moisture, fat and maximum carcass protein were evident in fish fed 1.75% threonine level. However, ash content did not affect body composition, except the 1.00% threonine level, which showed a significantly higher ash content value. Based on the above results, it is recommended that the diet for C. mrigala should contain threonine at 1.80 g 100 g^?1 dry diet, corresponding to 4.50 g 100 g^?1 dietary protein for optimum growth and efficient feed utilization.     

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Dietary arginine requirement of fingerling Indian major carp, Cirrhinus mrigala (4.20 ± 0.05 cm; 0.60 ± 0.02 g) was determined by conducting a 8‐week feeding trial with casein–gelatine‐based diets (400 g kg^?1 crude protein; 17.90 kJ g^?1, gross energy), containing crystalline amino acids with graded levels of l ‐arginine (10, 12.5, 15, 17.5, 20 and 22.5 g kg^?1, dry diet). Fish were randomly stocked, in triplicate groups, in 55‐L indoor polyvinyl flow through circular tanks and fed experimental diets at 5% of their body weight divided into two feedings at 08.00 and 16.00 hours. Live weight gain (321%) and feed conversion ratio (FCR 1.40) were significantly (P < 0.05) higher in fish fed diet containing 17.5 g kg^?1dietary arginine compared with other diets. Second‐degree polynomial regression analysis of live weight gain, FCR and protein efficiency ratio data indicated requirements for dietary arginine at 18.7, 18.4 and 18.3 g kg^?1 of the dry diet, respectively. Maximum carcass protein, and minimum moisture and fat contents were noticed at the requirement level. Carcass ash content remained insignificantly different among the treatments except at 17.5 g kg^?1 dietary arginine showing significantly higher ash content. Based on the above results, it is recommended that the diet for fingerling C. mrigala should contain arginine at 18.4 g kg^?1, dry diet, corresponding to 46 g kg^?1 dietary protein for optimum growth and efficient feed utilization.     

Dietary tryptophan requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

An 8‐week feeding trial was conducted to evaluate the effects of dietary tryptophan concentration on weight gain and feed efficiencies of fingerling Indian major carp, Cirrhinus mrigala. Six isonitrogenous (40% crude protein) and isocaloric (17.90 kJ g^?1) amino acid test diets containing casein, gelatin and l ‐crystalline amino acids with graded levels of l ‐tryptophan (0.06, 0.16, 0.26, 0.36, 0.46 and 0.56 g 100 g^?1 dry diet) were formulated. Fish (4.25±0.30 cm, 0.62±0.02 g) were randomly stocked in triplicate groups in 70 L (water volume 55 L) flow‐through (1–1.5 L min^?1) indoor circular tanks and fed experimental diets at 5% of their body weight/day in two feedings at 08:00 and 16:00 hours. Maximum live weight gain (277%), lowest feed conversion ratio (FCR) (1.50) and highest protein efficiency ratio (PER) (1.66) were measured at 0.36% dietary tryptophan. The relationship between dietary tryptophan levels and weight gain, FCR and PER data were described using second‐degree polynomial regression analysis indicating the tryptophan requirement at 0.42, 0.39 and 0.38 g 100 g^?1 of dry diet respectively. Whole body moisture decreased with increasing tryptophan up to 0.36%. Significantly (P<0.05) higher protein content was evident in fish fed diet containing 0.36% tryptophan. Body fat increased significantly (P<0.05) in fish fed with different tryptophan concentrations except those fed 0.36% tryptophan where a significantly lower fat content was noted. Significantly (P<0.05) higher ash content was reported at 0.06% and 0.16% tryptophan levels. Survival was 100% in fish fed all the diets except those fed 0.06% tryptophan. Based on the results, diets for fingerling C. mrigala should contain tryptophan at 0.38 g 100 g^?1 dry diet, corresponding to 0.95 g 100 g^?1 dietary protein for optimum growth and efficient feed utilization.     

Dietary methionine requirement of fingerling Indian major carp, Cirrhinus mrigala (Hamilton)

Indian major carp, Cirrhinus mrigala fingerling (3.85 ± 0.50 cm, 0.50 ± 0.02 g) were fed isonitrogenous and isocaloric diets (40% CP, 4.28 kcal g⁻¹, GE) containing casein, gelatin and crystalline amino acids with graded levels of L- methionine (0.50, 0.75, 1.00, 1.25, 1.50 and 2.00 g/ 100 g, dry diet) with 1.00% cystine fixed, to determine its dietary methionine requirement. A feeding trial was conducted in triplicate for six weeks. Diets were fed twice a day at 0800 and 1600 h at 5% of body weight/day. The ration size and feeding regime were worked out prior to the start of the feeding trial. Weight gain (158%) and food conversion ratio (1.45) were significantly (P < 0.05) higher in fish fed diet containing 1.00% methionine with 1.00% cystine fixed. Second degree polynomial regression analysis of the weight gain data indicated the dietary methionine requirement to be 1.20 g/100 g of dry diet, corresponding to 3.00% of dietary protein. Second degree polynomial regression analysis was also employed to determine the relationship between food conversion ratio (FCR) and dietary methionine levels which indicated that the best FCR occurred at approximately 1.20% dietary methionine level. Carcass composition of fish fed diet containing graded levels of methionine varied significantly (P < 0.05) except carcass ash content which showed insignificant (P > 0.05) differences among the dietary methionine levels. This revised version was published online in August 2006 with corrections to the Cover Date.     

Dietary thiamin and pyridoxine requirements of fingerling Indian major carp,Cirrhinus mrigala (Hamilton)

Two experiments were conducted to quantify the dietary thiamin (experiment I) and pyridoxine (experiment II) requirements of fingerling Cirrhinus mrigala for 16 weeks. In experiment I, dietary thiamin requirement was determined by feeding seven casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) with graded levels of thiamin (0, 0.5, 1, 2, 4, 8 and 16 mg kg^?1 diet) to triplicate groups of fish (6.15 ± 0.37 cm; 1.89 ± 0.12 g). Fish fed diet with 2 mg kg^?1 thiamin had highest specific growth rate (SGR), protein retention (PR), RNA/DNA ratio, haemoglobin (Hb), haematocrit (Hct), RBCs and best feed conversion ratio (FCR). However, highest liver thiamin concentration was recorded in fish fed 4 mg thiamin kg^?1 diet. Broken‐line analysis of SGR, PR and liver thiamin concentrations exhibited the thiamin requirement in the range of 1.79–3.34 mg kg^?1 diet (0.096–0.179 μg thiamin kJ^?1 gross energy). In experiment II, six casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) containing graded levels of pyridoxine (0, 2, 4, 6, 8 and 10 mg kg^?1 diet) were fed to triplicate groups of fish (6.35 ± 0.37 cm; 1.97 ± 0.12 g). Fish fed diet containing 6 mg kg^?1 pyridoxine showed best SGR, FCR, PR, RNA/DNA ratio, Hb, Hct and RBCs, whereas maximum liver pyridoxine concentration was recorded in fish fed 8 mg kg^?1 dietary pyridoxine. Broken‐line analysis of SGR, PR and liver pyridoxine concentrations reflected the pyridoxine requirement from 5.63 to 8.61 mg kg^?1 diet. Data generated during this study would be useful in formulating thiamin‐ and pyridoxine‐balanced feeds for the intensive culture of this fish.     

Evaluation of antibacterial activity and innate immune components in skin mucus of Indian major carp,Cirrhinus mrigala

The present work has been undertaken to analyse the antibacterial activity and innate immune components in the skin mucus of Indian major carp, Cirrhinus mrigala. Skin mucus was extracted separately in triple‐distilled water (TDW), 3% acetic acid (3% AA) or 0.1% trifluoroacetic acid (1% TFA). All mucus extracts exhibited different spectrum of the antibacterial activity against different groups of pathogenic bacteria. Protein profiling by polyacrylamide gel electrophoresis revealed a series of protein bands in the TDW extract, four major protein bands in the AA extract and two protein bands in the TFA extract. Tandem mass spectrometry analysis of distinct protein bands identified potential innate immune factors – histone H2A, histone H3, histone H4, haemoglobin, cofilin and nucleoside diphosphate kinase in the TDW extract, and ubiquitin and histone H2B isoforms in acidic extracts of skin mucus of C. mrigala. The presence of these innate immune molecules suggests that skin mucus play an important role in the protection of the fish against microbial invasion.     

Ontogenic changes in the digestive enzyme patterns and characterization of proteases in Indian major carp Cirrhinus mrigala

Digestive enzymes of Cirrhinus mrigala (Ham.) were studied during ontogenic development. Specific amylase activity was detected in first feeding fish. The enzyme activity decreased up to day‐18 and then it increased with the age of fish to reach the highest level on day‐34. Protease activity was 28.61 ± 8.90 mU mg protein^?1 min^?1 on day‐4 and increased with the age throughout the study period. Trypsin activity was 31.86 ± 1.12 mU mg protein^?1 min^?1 on day‐4. The activity decreased up to day‐10 and from day‐12 onwards increased up to day‐26. Chymotrypsin activity was 14.56 ± 2.74 mU mg protein^?1 min^?1on day‐4 and constantly increased up to day‐26. A significant increase in lipase activity was observed between days‐24 and 34. SDS‐PAGE and substrate SDS‐PAGE showed the diversity of protein (17.4–127.8 kDa) and protease activity bands (16.6–88.8 kDa) during ontogenesis. Soybean trypsin inhibitor, phenyl methyl sulphonyl fluoride, N‐α‐p‐tosyl‐l ‐lysine chloromethylketone and N‐tosyl‐l ‐phenylalanine chloromethylketone inhibited the protease activity up to 79.72–97.21, 65.55–94.83, 45.41–75.31 and 40.78–64.72%, respectively. Inhibition study in substrate SDS‐PAGE revealed the abundance of serine proteases and the presence of isoforms of trypsin and chymotrypsin. Ethylenediamine‐tetraacetate showed 5.56–22.78% inhibition of metal ion‐specific enzyme activity.     

Effect of soybean-meal-based diets on the growth and survival rate of the Indian major carp, Cirrhinus mrigala (Ham.)

The effect of varying dietary levels of defatted soybean meal on the growth and survival of mrigal, Cirrhinus mrigala (Hamilton) was investigated. In a feeding trial of 90 days, three experimental diets containing soybean meal at 200, 300 and 400 g kg^?1 level of incorporation were fed to quadruplicate groups of 10 fish each. The conventional feed used in India, consisting of a mixture of groundnut oil cake and rice bran in 1 : 1 ratio served as the control. Best growth in terms of percentage weight gain, specific growth rate, protein efficiency ratio (PER), feed conversion ratio and survival rate was obtained for the test diet with 354 g kg^?1 crude protein and with 400 g kg^?1 soybean meal inclusion level. However, no statistical significant difference was observed between the three soybean‐based diets, except for PER and survival rate. Soybean meal is an easily available, acceptable and cost‐effective protein source in formulated feeds for Indian major carps. The results of the present study indicate that a diet of 350 g kg^?1 overall protein with soybean meal included at 400 g kg^?1 can elicit good growth response and survival in mrigal.     

Nutritional requirement of ascorbic acid by Indian major carp,Cirrhina mrigala,during early growth

Using synthetic diets, hatchlings of Cirrhina mrigala (obtained through induced breeding) were fed graded levels of ascorbic acid: 0, 60, 300, 600, 900, and 1200 mg/kg diet for 240 days. Studies on survival, growth and morphological changes of these fishes are reported. Statistical analysis of growth, deformity and survival rates at these levels suggested an optimum requirement of 650–750 mg ascorbic acid/kg diet in this early part of the life of the fish. Avitaminosis resulted in poor growth, high mortality rate (up to 42%), severe haemorrhages, fin necrosis, increased pigmentation and spinal flexures. 60 mg and 300 mg ascorbic acid/kg dietary levels also resulted in poor survival and growth rates. Skeletal deformities were also observed in some of these fish.     

Environmental effects of common carp Cyprinus carpio (L.) and mrigal Cirrhinus mrigala (Hamilton) as bottom feeders in major Indian carp polycultures

A polyculture experiment with the large carp rohu, catla and either mrigal or common carp (as cash crop fish), and the small indigenous fish punti (as food for the farmer's family) was carried out at Bangladesh Agricultural University, Mymensingh. The main objectives were to compare polycultures of large carp in which the bottom feeder is either the native mrigal or the exotic common carp, and to assess the effects of adding the small indigenous species punti to those polycultures. The results of fish–fish interactions and overall fish production have already been reported. The present paper presents the effects on the water quality, and discusses fish–environment interactions. The main conclusions are: time changes in the pond environment were stronger than fish composition effects. The main practice affecting water quality was liming, that incresed alkalinity, pH and water transparency and decreased ammonia. Rain affected photosynthesis and the match‐mismatch of the two steps of nitrification. The more that bottom feeding fish species disrupt the mud bottom, the stronger their effects on pond environment. Common carp produce the strongest disruption of the mud bottom, followed by punti and then by mrigal. Mud disruption produced by common carp leads to a stronger liming effect, nutrient release into the water, and provides more particles that rain‐floods wash out, facilitating the mismatch of the two steps of nitrification, and increased phosphorus adsorption into the mud bottom. Mud disruption by punti is only enough to improve the liming effect. Mud disruption by mrigal is the least, hence less particles are resuspended, nitrification is not affected during floods and relatively more phosphate remains in the water available for photosynthesis. The bottom feeder common carp can be seen not only as a target‐cultured fish but also as a management tool. Farmers can get double benefit in introducing common carp in the ponds as it enhances the effectiveness of lime application and increases the availability of nutrients to phytoplankton. Through the manipulation of species in the polyculture alone, farmers can maintain the environment better and also reduce input costs.     

The effect of common carp, Cyprinus carpio (L.) and mrigal, Cirrhinus mrigala (Hamilton) as bottom feeders in major Indian carp polycultures

A polyculture experiment with the large carp rohu, Labeo rohita (Hamilton), catla, Catla catla (Hamilton) and either mrigal, Cirrhinus mrigala (Hamilton) or common carp, Cyprinus carpio (L.) (as cash crop fish), and the small indigenous fish punti, Puntius sophore (Hamilton) (as food for the small‐scale farmer family) was carried out at the Field Laboratory of the Faculty of Fisheries, Bangladesh Agricultural University, Mymensingh. The main objective was to compare polycultures of large carp in which the bottom feeder is either the native mrigal or the exotic common carp. Secondary objectives were to assess the effects of adding the small indigenous species punti to polycultures of large carp, and to compare the effects of mrigal and common carp on punti production and reproduction. It was found that (i) common carp damaged embankments, had no effect on catla, improved rohu performance by 50% and total fish production by 20%; (ii) punti addition did not affect rohu, catla and total yield, improved mrigal performance by 50%, and decreased common carp performance by 20%; and (iii) punti was not affected either by common carp or by mrigal. However, its performance was not satisfactory, probably owing to frequent netting, which might have hindered growth and breeding. In spite of the embankment damage caused by common carp, this bottom feeder seems to be more promising than mrigal, because it leads to higher fish production. The addition of punti to the large carp polyculture is a viable proposition, as it does not reduce cash crop production, and might be a good food source for a small‐scale farmer's family.     

Reproduction phase-related variations in the GnRH immunoreactive fibers in the pineal of the Indian major carp Cirrhinus mrigala (Ham.)

We studied GnRH immunoreactivity in the pineal gland of the Indian major carp Cirrhinus mrigala during different phases of reproductive cycle. In the resting phase (December–January), GnRH immunoreactive (-ir) fibers were organized as paired fascicles above the posterior commissure that ascend in the stalk and distribute widely in the pineal gland. The GnRH-ir fiber density significantly declined (P<0.001) during the preparatory phase (February–April) and the fibers disappeared thereafter. While no GnRH fibers were seen during the prespawning (May–June) and spawning (July–August), isolated GnRH-ir fibers reappeared in the postspawning phase. Since no GnRH cell bodies were detected in the pineal, these GnRH-ir fibers seem to be of central origin. The results reveal a distinct reciprocal relationship between the GnRH immunoreactivity in the pineal and the status of the ovarian maturity; the fibers appeared in the pineal only during the period of ovarian quiescence. While the functional significance of these cyclic changes in GnRH is yet to be determined, we suggest that the decapeptide may serve as a transmitter of central origin that modulates the activity of the pineal gland.     

Dietary methionine requirement of Indian major carp fry,Cirrhinus mrigala (Hamilton) based on growth,feed conversion and nitrogen retention efficiency

This study was aimed at quantifying methionine requirement of Indian major carp fry, Cirrhinus mrigala (2.2 ± 0.2 cm; 0.19 ± 0.02 g) by conducting a 12‐week feeding trial. Casein–gelatine‐based isonitrogenous (40 g 100 g^?1 crude protein) and isoenergetic (15.42 kJ g^?1 DE) amino acid test diets were prepared to contain six levels of l ‐methionine (1.1, 1.3, 1.5, 1.7, 1.9 and 2.1 g 100 g^?1 dry diet) at a fixed level of cysteine (0.85 g 100 g^?1 dry diet) and fed to apparent satiation thrice daily to triplicate groups of fish. When absolute weight gain (g per fish), feed conversion ratio, protein deposition (g per fish) and nitrogen retention efficiency data were subjected to broken‐line and second‐degree polynomial regression analysis, 95% of the plateau of above parameters was achieved at dietary methionine concentrations between 1.60 and 1.69 g 100 g^?1 dry diet or 0.10 to 0.11 g methionine kJ^?1 DE, corresponding to 4.1–4.22 g 100 g^?1 protein or 0.44–0.47 g methionine kJ^?1 DE. Based on these results, dietary methionine requirement of fry C. mrigala is recommended 1.60–1.69 g 100 g^?1 diet or 0.10–0.11 g methionine kJ^?1 DE.     

Nutritional deficiency disease in an Indian major carp, Cirrhina mrigala Hamilton, due to avitaminosis C during early growth

Abstract. Experiments with a purified vitamin test diet on newly hatched Cirrhina mrigala Hamilton, a major carp of great piscicultural importance, are reported. Experimentally induced vitamin C deficiency for 330 days was followed by recovery treatment for 30 days. Retarded growth and greater mortality accompanied by structural deformities such as lordosis and scoliosis, increasing with prolonged deficiency, were recorded. Haematological and haematopoietic studies revealed hypochromic macrocytic anaemia accompanied by anisocytosis. Correlated studies on erythropoiesis revealed increased reticulocytes and an overall decrease in small lymphoid haemoblasts. Total and differential leucocyte counts revealed leucopenia and thrombopenia accompanied by an increase in neutrophils and eosinophils. Recovery treatment halted and in some cases even compensated these pathophysio-logical changes to some extent.     

Laboratory and field investigations on the effect of scheduled meal timings on growth performance and nutrient retention in an Indian major carp,Cirrhinus mrigala (Ham) fingerlings: Effect on nitrogen retention and excretion of metabolites

To investigate the effect of scheduled meal timings on growth performance in Cirrhinus mrigala fingerlings, two experiments were conducted. The first experiment was conducted under laboratory conditions and the fish were submitted to schedule meal timings (at 08:00, 12:00,16:00, 20:00, 00:00 and 04:00). A control on continuous feeding was also maintained. ANOVA had revealed a significant (p < .05) increase in live weight gain (g), growth per cent gain in body weight, specific growth rate, PER, GPR, GER and APD (%) values in fingerlings fed between 12:00 and 16:00 hours. A decline in growth parameters, nutrient retention and an increase in FCR values were observed in the group fed at 20:00, 00:00 hours and also in the control group. Studies have further revealed that meal timings had also significantly (p < .005) affected protein digestibility, nitrogen retention and excretion of metabolites . Fish carcass composition had significantly (p < .05) higher accumulation of protein (14.82 ± 0.032), fat (5.51 ± 0.006) and energy (5.95 ± 0.004) in the group fed at 16:00 hours. The second experiment was conducted under field conditions and the fish were submitted to schedule meal timings (at 08:00, 12:00, 16:00 and 20:00). A control on continuous feeding was also maintained. Significantly (p < .05) higher values in growth parameters were observed in the group fed at 16:00 hours and lower values in the group fed at 20:00 hours and also in controls. Water quality, nutrients and productivity status of ponds revealed favourable levels and appears to have been affected by meal timings. Thus, in C. mrigala, timings of food intake can serve to optimize the utilization of ingested calories.     

Characterization of carboxylesterase in skin mucus of Cirrhinus mrigala and its assessment as biomarker of organophosphate exposure

Presence of carboxylesterase (CbE) activity in the skin mucus of Cirrhinus mrigala was investigated. CbE activity in skin mucus showed higher substrate preference for α-naphthyl acetate over p-nitrophenyl acetate. Four CbE isozymes—CbE-1, CbE-2, CbE-3, and CbE-4 were observed in skin mucus during zymography. The isozyme CbE-4 was characterized as typical serine esterase, whereas CbE-1, CbE-2, and CbE-3 were identified as sulphhydryl group-dependent serine esterases. In vitro treatment of skin mucus with the organophosphorus insecticide, Nuvan^® showed strong inhibition of CbE activity. In vivo exposure of the fish to sublethal test concentrations (5 and 15 mg/l) of the insecticide also revealed significant inhibition of CbE activity in mucus. After the cessation of exposure, CbE activity recovered to its control level during the recovery periods. Thus, CbE activity in skin mucus could be considered a biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity.     

Secondary stress responses in Indian major carps Labeo rohita (Hamilton), Catla catla (Hamilton) and Cirrhinus mrigala (Hamilton) fry to increasing packing densities

Glycogen content and metabolic enzyme activities viz. lactate dehydrogenase (LDH), malate dehydrogenase (MDH), aspartate amino transferase (AST) and alanine amino transferase (ALT) in Indian major carps, Labeo rohita, Catla catla and Cirrhinus mrigala, were investigated after a 6 h transportation trial to compare the species‐specific variation and the effect of increased packing density on the metabolism. Fish (45±5 mm, 0.5±0.1 g) were packed in three densities (100, 150 and 200 L^?1) for the experiment, and 12 specimens of each species were randomly sampled from all the treatments at the end of transportation. The glycogen content of L. rohita ingerlings decreased significantly (P<0.05) with increasing packing density. The activities of enzymes LDH, MDH, AST and ALT showed a rising trend with increasing packing density in all the three species. Species‐specific differences were observed in various tested parameters at the lowest packing density (100 fry L^?1). Alanine amino transferase and LDH activities were significantly (P<0.05) lower in C. mrigala as compared with the other two species. However, glycogen reserves and MDH activity were not significantly different (P>0.05) among the species. The present study reveals that the optimum packing density for Indian major carp fry (100 fry L^?1) for transportation up to 6 h and metabolic regimes are species specific during transportation.     

Identification and partial characterization of potential probiotic lactic acid bacteria in freshwater Labeo rohita and Cirrhinus mrigala

Carps are the most diversified freshwater fish belonging to family Cyprinidae. Numerous probiotic and pathogenic lactic acid bacteria (LAB) have been characterized from carps. However, the diversity of these ecologically important bacteria is entirely unknown in freshwater fish of Pakistan. The present study aimed to characterize and identify the lactic acid bacteria from two carps viz. Laboe rohita and Cirrhinus mrigala and determine their antagonistic activity. Seventeen bacterial isolates were purified from the gastrointestinal tract and gills of these fish and characterized morphologically. Initially, seven isolates were screened as LAB using agar supplemented with CaCO₃. Subsequently, only two isolates CILB2 and RIL10 were selected as LAB after high‐performance liquid chromatography analysis for lactic acid production. Isolates CILB2 and RIL10 were genetically identified as Enterococcus faecalis and Weissella sp., respectively after 16S rRNA gene sequence analysis. Both strains exhibited significant antagonistic activity against common fish pathogens Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli. Enterococcus faecalis CILB2 and Weissella sp. RIL10 were also found negative for haemolysis and gelatinase activities and were sensitive to ampicillin, amoxicillin, doxycycline, erythromycin, chloramphenicol and co‐trimoxazole antibiotics. The identified LAB strains may further be investigated for their potential probiotic application in fish feed and food preservation techniques.     

Total sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton)

The quantitative dietary sulphur amino acid requirement of the Indian major carp, Labeo rohita (Hamilton), was determined by conducting a growth study. The experimental diets contained 400 g crude protein kg⁻¹ from casein, gelatine and supplemental crystalline amino acids. Diets containing six graded levels of methionine (3.2, 6.5, 9.0, 11.5, 14 and 16.5 g kg⁻¹) with a constant level of cystine (1.4 g kg⁻¹) were formulated and fed to triplicate groups of Labeo rohita fingerlings twice a day to satiation for 60 days. The optimum dietary requirement for methionine was estimated using the break-point regression analysis at 11.5 g kg⁻¹ of diet or 28.8 g kg⁻¹ of dietary protein. Thus the total sulphur amino acid (Met + Cys) requirement was determined to be 12.9 g kg⁻¹ of diet or 32.3 g kg⁻¹ of protein. Higher survival, specific growth rate and food conversion efficiency values were observed for fish fed the diet containing optimum levels of sulphur amino acids.