Hematological and plasma chemistry of Indian major carp,Labeo rohita (Hamilton, 1822)

Hematological and plasma chemistry indices are simple and essential diagnostic tools for monitoring the physiological and health status of fish. Aim of the present study was to obtain reference values for the hematological and plasma chemistry of wild populations of Labeo rohita captured in a freshwater pond between July 2008 and June 2010. These reference values and the mean were evaluated according to sex and season. In summer, the red blood cells (1.84 × 106 38 per cubic mm), haemoglobin (8.52 gm dl^?1) and haematocrit (31.49%) were highest in males, whereas the maximum values for white blood cells (5.635 × 103 40 per cubic mm) were found in females, however, no significant variation of mean corpuscular volume (MCV) or mean corpuscular haemoglobin (MCH) was observed between sexes. Various blood parameter levels between the sexes in summer were notably different from those measured in other seasons except for MCH and MCHC values (p < 0.05). Compared to most teleosts, the L. rohita has similar mean values for PCV and Hb. Throughout summer the glucose (76.0 mg dl^?1), lipid (3.41 gm dl^?1) and cholesterol (145.0 mg dl^?1) levels were highest. In spring the plasma protein levels were higher in males, but higher in winter for females. Consequently, the seasons are key factors when using blood parameters as biomarkers for environmental alterations.     

Dietary valine requirement of Indian major carp, Labeo rohita (Hamilton) fry

Dietary valine requirement of Indian major carp, Labeo rohita Hamilton, fry (3.0 ± 0.02 cm, 0.16 ± 0.03 g) was determined using dose‐response method. Fishes were fed six isonitrogenous [40% crude protein (CP)] and isocaloric (4.28 kcal g^?1, Gross Energy (GE)) amino acid test diets containing casein, gelatin, and l ‐crystalline amino acids with graded levels of valine (0.75, 1.00, 1.25, 1.50, 1.75, and 2.00% dry diet) at 5% body weight for 6 weeks in triplicate groups twice a day at 07.00 and 17.30 hours. Live weight gain (158.52%), feed conversion ratio (FCR, 1.70), specific growth rate (SGR, 2.25), and protein efficiency ratio (PER, 1.46) were significantly (P < 0.05) higher in fish fed a diet containing 1.5% of the dietary valine (diet IV). Second‐degree polynomial regression analysis of the live weight gain and FCR data indicated the dietary valine requirement at 1.63 and 1.5% of the dry diet, corresponding to 4.0 and 3.75% of dietary protein. Maximum carcass protein, minimum moisture, and fat were recorded at 1.5% of the dietary valine level, except carcass ash, which remained constant throughout the treatments. No mortality was observed during the entire length of the feeding trial. On the basis of FCR and protein deposition data, it is recommended that dietary valine inclusion at 1.5% of dry diet, corresponding to 3.75% of dietary protein, is optimal for the growth of L. rohita fry.     

High temperature acclimation alters upper thermal limits and growth performance of Indian major carp,rohu, Labeo rohita (Hamilton, 1822)

Increase in water temperature due to anthropogenic and climatic changes is expected to affect physiological functions of fish. In this study, we determined high temperature tolerance (CTmax) of a common aquacultured Indian major carp, rohu, Labeo rohita fingerlings (15.96 ± 0.72 g BW, 11.56 ± 0.42 cm TL) followed by acclimatization at three temperatures (30, 33, 36 °C). To determine the CTmax, we analyzed the major hemato-biochemical indices - hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC), blood glucose levels, and erythrocytic nuclear abnormalities (ENAs) and erythrocytic cellular abnormalities (ECAs) of peripheral erythrocytes in the fish sampled at the start and end point at each acclimated temperature. Significantly decreased CTmax of the fish was found at 36 °C compared to 30 °C and 33 °C. The fish in the highest (36 °C) temperature were found with significantly lower Hb and RBC content and significantly higher WBC and blood glucose levels than that of the fishes in the lowest (30 °C) temperature both at the start and end points. The highest frequencies of ENAs and ECAs were found in the highest (36 °C) temperature group compared to the lowest (30 °C) temperature group at both the points. We also evaluated growth performance of the rohu fingerlings reared in the three temperatures for 60 days. The growth parameters - final weight gain, percent weight gain and specific growth rate were the highest at 33 °C and the lowest at 36 °C. The present study revealed that the highest temperature (36 °C) tested here may be hazardous to rohu and the temperature should be kept below 36 °C in the aquaculture setting to avoid physiological damage and growth and production loss to the fish.     

De novo assembly,transcriptome characterization and marker discovery in Indian major carp,Labeo rohita through pyrosequencing

Genetica - Labeo rohita, one of the Indian major carps, is the most popular culture species in Indian subcontinent due to its consumer preference and delicacy. A selective breeding program for...     

Effect of endotoxin on the immunity of Indian major carp, Labeo rohita

Endotoxin, a lipopolysaccharide component of outer cell wall membrane of the Gram-negative bacteria is a factor responsible for a number of biological effects including immunostimulatory activities in different animal species including fish. In this study, L. rohita yearlings of weight ranging from 80 to 100g were injected intraperitoneally with 0.5, 1, 2, 5, 10 and 20 EU/fish dose of endotoxin to find out its effect on the immunity. The L. rohita yearlings were found to resist the endotoxin dose up to 20 EU/fish and at the lower doses, i.e., at 1 and 2 EU/fish; it acted as an immune potentiator. Different serum and immune parameters like protein, globulin, lysozyme, respiratory burst activity, myeloperoxidase activity, natural agglutination titre were found to be significantly high (p<0.01) at a dose of 1 EU/fish. While at 10 and 20 EU/fish, most of these parameters were lower thereby indicating the immuno-suppressive nature of the endotoxin at these higher doses.     

Isolation and identification of yolk proteins in Indian major carp,Labeo rohita

Two yolk proteins (YP1 and YP2) from the ovaries of Indian major carp, Labeo rohita were isolated by gel filtration and partially characterized by the use of hydroxyapatite ultrogel column in conjunction with native PAGE. On native PAGE YP1 gave a single protein band, whereas YP2 of gel filtration revealed the contamination of YP1, which was removed by adsorption chromatography on hydroxyapatite ultrogel and then the YP2 was the purified one as judged by electrophoresis. Both YP1 and YP2 also stained for lipid and contained alkalilabile phosphorus. Therefore, both yolk proteins were lipophosphoprotein. The molecular weights of YP1 and YP2 were 620 kDa and 225 kDa respectively as determined by gel filtration on Sepharose 4B. When YP1 and YP2 were compared in relation to some physicochemical characteristics with yolk proteins of other oviparous vertebrates including fish, they were lipovitellin like. Antiserum to YP2 crossreacted with YP2 and vitellogenin suggesting that YP2 was the cleaved product of vitellogenin. Anti-YP2 antiserum was not crossreacted with native YP1, whereas reduced and/or denatured YP1 was crossreacted indicating the presence of antigenic determinants in the inner core region of YP1 polypeptide.     

Randomly amplified polymorphic DNA analysis of four different populations of the Indian major carp, Labeo rohita (Hamilton)

The level of genetic variation provides the raw material for selective improvement of a stock. Random amplified polymorphic DNA (RAPD) assay was used to assess the genetic variation in three rivers: the Halda, the Jamuna and the Padma as well as in one hatchery population of the commercially important Indian major carp, Labeo rohita. RAPD markers were amplified from DNA samples of 35 fish from each of the four populations using six decamer random primers. The polymorphic loci proportions were 0.33, 0.28, 0.28 and 0.26 and Nei's gene diversity values were 0.06, 0.07, 0.06 and 0.05 for the Halda, the Jamuna, the Padma and the hatchery populations, respectively. The pairwise population differentiation (F_ST) values indicated a low level of genetic differentiation between the population pairs. From the unweighted pair group method of arithmetic mean (UPGMA) dendrogram based on Nei's genetic distances a correlation between genetic affinities and geographical area was found. The populations were segregated into two groups: the Halda in one group and the Jamuna, the Padma and the hatchery in another group. Overall, the RAPD technique can be introduced as a tool in the population genetics of the rohu fish to provide information on their genetic stock structure.     

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.     

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.     

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.     

Revealing liver specific microRNAs linked with carbohydrate metabolism of farmed carp,Labeo rohita (Hamilton, 1822)

The role of microRNA in gene regulation during developmental biology has been well depicted in several organisms. The present study was performed to investigate miRNAs role in the liver tissues during carbohydrate metabolism and their targets in the farmed carp rohu, Labeo rohita, which is economically important species in aquaculture. Using Illumina-HiSeq technology, a total of 22,612,316; 44,316,046 and 13,338,434 clean reads were obtained from three small-RNA libraries. We have identified 138 conserved and 161 novel miRNAs and studies revealed that miR-22, miR-122, miR-365, miR-200, and miR-146 are involved in carbohydrate metabolism. Further analysis depicted mature miRNA and their predicted target sites in genes that were involved in developmental biology, cellular activities, transportation, etc. This is the first report of the presence of miRNAs in liver tissue of rohu and their comparative profile linked with metabolism serves as a vital resource as a biomarker.     

Non-specific immune parameters of brood Indian major carp Labeo rohita and their seasonal variations

Different non-specific immune parameters and their seasonal changes in brood Indian major carp Labeo rohita reared in two major freshwater aquaculture regions of India viz. West Bengal and Orissa were investigated. It was undertaken for 2 consecutive years and included three main seasons of a year such as summer (March-May), rainy (July-September) and winter (November-January). Total serum protein, albumin and globulin levels were not significantly different throughout the year (p>0.01). Serum lysozyme and myeloperoxidase activities were lower (7.26+/-0.87mg/ml and, 0.54+/-0.11 OD, respectively) in winter as compared to any other season of the year. The bacterial agglutination titer was higher (p<0.01) in the rainy season (8.70+/-1.70) compared to summer and winter seasons (3.40+/-0.60 and 4.00+/-0.89, respectively). Haemagglutination and haemolytic activities did not vary (p>0.01) throughout the year. In blood smears, lymphocyte percentage was higher (75-80%) as compared to those of neutrophil (10-15%) and monocytes (5-10%) but eosinophilic granulocytes were present only in few cases. The differential leucocyte count did not vary significantly (p>0.05) in any season. This study indicated that certain non-specific immune parameters of this species can be modulated at certain times of the year.     

Efficacy of water hyacinth compost in nursery ponds for larval rearing of Indian major carp,Labeo rohita

The use of water hyacinth compost as manure in nursery ponds for larval rearing of Indian major carp, Labeo rohita was studied. Better performance with regard to growth and survivability of the larvae was recorded in the ponds treated with water hyacinth compost (Group A) than in either the ponds treated with inorganic fertilizers (Group B) or the ponds where no treatment was applied (Group C). The average percentages of survival in Groups A, B and C were 14.3, 11.2 and 5.0, respectively. Thus about 186% increase in the survivability was recorded in the ponds treated with water hyacinth compost (P < 0.01), whereas the increase was about 124% in the ponds treated with only inorganic fertilizers, when compared with the ponds which were without any treatment.     

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.     

Exploring the gut microbiota composition of Indian major carp,rohu (Labeo rohita), under diverse culture conditions

Gut microbiota of freshwater carps are often investigated for their roles in nutrient absorption, enzyme activities and probiotic properties. However, little is known about core microbiota, assembly pattern and the environmental influence on the gut microbiota of the Indian major carp, rohu. The gut microbial composition of rohu reared in different culture conditions was analysed by 16S rRNA amplicon sequencing. There was variation on gut microbial diversity and composition. A significant negative correlation between dissolved oxygen content (DO) and alpha diversity was observed, thus signifying DO content as one of the key environmental factors that regulated the diversity of rohu gut microbial community. A significant positive correlation was observed between phosphate concentration and abundance of Actinobacteria in different culture conditions. Two phyla, Proteobacteria and Actinobacteria along with OTU750868 (Streptomyces) showed significant (p < 0.05) differences in their abundance among all culture conditions. The Non-metric multidimensional scaling ordination (NMDS) analysis using Bray-Curtis distances, showed the presence of unique gut microbiota in rohu compared to other herbivorous fish. Based on niche breadth, 3 OTUs were identified as core generalists, persistent across all the culture conditions whereas the specialists dominated in the rohu gut microbiota assembly. Co-occurrence network analysis revealed positive interaction within core members while mutual exclusion between core and non-core members. Predicted microbiota function revealed that different culture conditions affected the metabolic capacity of gut microbiota of rohu. The results overall indicated the significant effect of different rearing environments on gut microbiota structure, assembly and inferred community function of rohu which might be useful for effective manipulation of gut microbial communities of rohu to promote better health and growth under different husbandry settings.     

The effect of dietary immunomodulation upon Edwardsiella tarda vaccination in healthy and immunocompromised Indian major carp (Labeo rohita)

In order to determine the impact on disease resistance of four dietary immunomodulators viz., beta-1,3 glucan, levamisole, vitamins C and E, in an important farmed Indian major carp species, rohu (Labeo rohita Ham.), fish were fed diets containing various levels of these substances during a 60 day trial. Aflatoxin B1 (AFB,) at 125 mg kg(-1) body weight was injected intraperitoneally (i.p.) into fish to induce an immunosuppressive state on the first day of the experiment in some individuals. The fish were vaccinated against formalin-killed Edwardsiella tarda vaccine on day 30 of the experiment. Specific immunity, as measured by bacterial agglutination titre and disease resistance against E. tarda, was determined at the end of the trial. The results demonstrate that all the four immunomodulators were capable of significantly (P<0.05) increasing specific immunity and reducing mortality in immunocompromised fish but failed to enhance specific immunity and protection in healthy fish. The increased bacterial agglutination titre by beta-1,3 glucan, and reduced mortality losses by both beta-1,3 glucan and levamisole were marked in healthy vaccinated fish compared with their controls. Similarly, all four substances significantly reduced the mortality rates in immunocompromised and healthy unvaccinated fish. Out of these four substances, glucan was recorded to be the most effective immunomodulator in rohu. The present results suggest that the introduction of these substances into the diet of fish grown in farms under immunosuppressive/stressful conditions could increase their resistance to infection by reducing mortality rates and offer economic benefits.     

Carcass traits of different marketable sizes of rohu,Labeo rohita (Hamilton, 1822)

The present study was carried out to investigate the carcass traits of farmed freshwater Labeo rohita in relation to body size as important information to calculate yield and/or support for machine/manual processing. For this purpose ninety specimens were collected for nine weight groups from 600 to 1100 g, each group with a 100 g increment. Specimens were collected from grow‐out culture ponds of the Central Institute of Freshwater Aquaculture (CIFA), Bhubaneswar. The carcass evaluation technique was followed to evaluate carcass yield, offal yield and carcass cutability. Head size percentage of rohu increased as the body mass increased, which in turn increased the processing yield. Dry matter and ether extract percentage also increased with the body mass; however, moisture percentage showed a decreasing trend. The fore cut showed a distinctly higher ether extract percentage followed by the middle and posterior cuts. There was a good correlation development to describe the relationship between yield traits and body weight.     

Demonstration of two forms of vitellogenin in serum of estradiol-17 beta-treated Indian major carp, Labeo rohita.

Vitellogenin (Vg) synthesis was induced in the male and non-vitellogenic female Rohu, the Indian major carp, by estradiol-17 beta(E2) where effect was more in female. A crude preparation of Vg was isolated in the second peak after gel filtration on Ultrogel AcA 34 from the sera of vitellogenic female Rohu and E2-treated male and female Rohu. Estimation of alkali-labile phosphorus was shown to be used as an index of Vg. Native-PAGE analysis has revealed the presence of two forms of Vg (Vg1: 430,000 dalton and Vg2:240,000 dalton) in Vg fraction obtained after gel filtration as well as in the sera of E2-treated male and female Rohu. Immunological cross-reaction studies between antiserum to yolk protein and Vg fractions as well as the sera from E2-treated male and female Rohu further indicates the presence of two precipitin lines (not clearly visible as the two lines fused to form a thick line) suggesting the occurrence of two forms of Vg in the Rohu.     

Family association between immune parameters and resistance to Aeromonas hydrophila infection in the Indian major carp, Labeo rohita

Seven innate immune parameters were investigated in 64 full-sib families (the offspring of 64 sires and 45 dams) from two year-classes of farmed rohu carp (Labeo rohita). Survival rates were also available from Aeromonas hydrophila infection (aeromoniasis) recorded in controlled challenge tests on a different sample of individuals from the same families. Due to strong confounding between the animal additive genetic effect and the family effects (common environmental + non-additive genetic), reliable additive (co)variance components and hence heritabilities and genetic correlations could not be obtained for the investigated parameters. Therefore, estimates of the association of challenge test survival with the studied immune parameters were obtained as product moment correlations between family least square means. These correlations revealed statistically significant (p < 0.05) negative correlations of survival with bacterial agglutination titre (−0.48), serum haemolysin titre (−0.29) and haemagglutination titre (−0.34); and significant positive correlation with ceruloplasmin level (0.51). The correlations of survival to aeromoniasis with myeloperoxidase activity, superoxide production and lysozyme activity were found to be not significantly different from zero (p > 0.05). Assuming that the negatively correlated candidate traits are not favourable as indirect selection criteria, the results suggest that ceruloplasmin level could potentially be a marker for resistance to aeromoniasis in rohu. The use of this immune parameter as an indirect selection criterion for increased resistance to aeromoniasis in rohu will, however, require that the parameter shows significant additive genetic variation and a significant genetic correlation with survival. Further studies are therefore needed to obtain a reliable heritability estimate for ceruloplasmin and its genetic correlation with survival from aeromoniasis.