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...     

Isolation and characterization of polymorphic microsatellites in Labeo rohita and their cross‐species amplification in related species

The major Indian carps namely rohu (Labeo rohita), catla (Catla catla), mrigal (Cirrhinus mrigala) and calbasu (Labeo calbasu) are important freshwater species of the Indian subcontinent constituting over 65% of the fish produce. In the present study, isolation of 12 microsatellite loci from rohu has been reported. Cross‐species amplification in related carps and their implication in population genetic studies as well as selective breeding program were discussed.     

Electrophoresis of haemoglobin in single erythrocytes of Labeo umbratus (Smith) and Labeo capensis (Smith)

Electrophoresis of haemoglobin in single erythrocytes of Labeo umbratus and Labeo capensis showed that haemoglobin behaved in three distinct ways in both species. The majority of cells contained haemoglobin which moved as a broad band towards the anode. In a few cells haemoglobin moved towards both the anode and the cathode and in even fewer cells, haemoglobin moved as a sharp band to either the cathode or anode. These results are discussed in relation to other observations.     

Aspects of reproductive biology of Labeo horie Heckel (Pisces: Cyprinidae) in Lake Chamo, Ethiopia

Labeo horie Heckel is increasingly becoming commercially important in Lake Chamo but its reproductive biology in the lake is not well enough to guide its management. Sex ratio, breeding season, length at first maturity and fecundity of L. horie were studied from 1197 fish samples collected between August 1998 and October 1999 in Lake Chamo, Ethiopia. The sex ratio was significantly different (χ ² = 12.12, P < 0.001). The peak‐breeding period was during the rainy months of March to May, during which time more than 90% of both sexes had ripe gonads. The size at maturity ( L _m50 ) of males was 52 cm while the L _m50 of females was 62 cm. The smallest ripe male in the catch was 46.7 cm and weighed 890 g while the smallest ripe female caught was 49.5 cm and weighed 1145 g. The weight of ripe ovaries ranged from 54.3 g to 991.8 g and contained between 68,400 and 1,320,400 eggs. Relative fecundity ranged between 60 and 290 eggs per g of body weight. The relationships between fecundity and total length and between fecundity and total weight were curvilinear, while the relationship between fecundity and ovary weight was linear. L. horie conformed to the general pattern of reproduction in a tropical environment where peak‐breeding activity occurred during the rainy season. In order to protect the spawning population, fishing pressure should be minimized during breeding time at the shallower littoral regions.     

Blood respiratory properties of the mudsucker fish Labeo capensis (Smith)

For Labeo capensis (Smith) the increase in blood organic phosphates (ATP, GTP) in winter fish exceeds the increase in summer fish. Blood chlorides, blood osmotic pressure, hematocrit and hemoglobin concentration decrease significantly in winter fish when compared to summer fish. Four effectors, viz. temperature, organic phosphates, H+ and Cl-, could lower the oxygen affinity of whole blood, especially in summer fish.     

Fish Cholinesterases as Biomarkers of Sublethal Effects of Organophosphorus and Carbamates in Tissues of Labeo Rohita

Organophosphates and carbamates are major agrochemicals that strongly affect different neuroenzymes and the growth of various fish species. Here, we study the effect of sublethal concentrations of profenofos and carbofuran on the activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) and the associated health risk in fish. Labeo rohita fingerlings were exposed to three sublethal concentrations of profenofos and carbofuran. The minimum cholinesterase activities in the brain, gills, muscle, kidney, liver, and blood were after exposure to profenofos (0.06 mg/L). The minimum AChE and BuChE activities in the brain, gills, muscle, kidney, liver, and blood were after exposure to carbofuran (0.28 and 0.198 mg/L). Exposure to both types of pesticides affected the functions of these organs, including metabolism and neurotransmission, to various extents at different exposure concentrations. These findings suggest that they are required to be properly monitored in the environment, to reduce their toxic effects on nontarget organisms     

Food and feeding habits of Labeo niloticus (Pisces,Cyprinidae) in Jebel Aulia Reservoir,Sudan

Basic knowledge on the feeding ecology of one of the common and commercially important fish species in Jebel Aulia Reservoir is provided.The structure of the feeding apparatus indicates that Labeo niloticus is a bottom feeder, depending on soft and decayed vegetation, organic debris and whatever small organisms found within. However, juveniles and fry are prone to explore all layers and depths of the river selectively for plankton. There is little evidence of seasonal selection of food. Changes in diet quality are governed by the availability of type of food. Variability of feeding activity is connected with climate and breeding season.     

Metal contamination and human health risk associated with the consumption of Labeo rosae from the Olifants River system,South Africa

The Olifants River, a tributary of the Limpopo River system, is one of the most polluted rivers in South Africa. In May 2011 the concentrations of metals in fish muscle tissue from two impoundments, Loskop and Flag Boshielo dams, on the Olifants River were measured and a human health risk assessment conducted to investigate whether it was safe to consume Labeo rosae from these impoundments. Labeo rosae is one of the most common pan fish in these impoundments and is readily available to rural communities. Metals are accumulating in the muscle tissue of L. rosae even although the fish populations appear to be healthy. At Loskop Dam all L. rosae analysed exceeded the recommended hazard quotient (HQ) of 1 for antimony, and less than 50% exceeded that for lead. At Flag Boshielo Dam, the recommended HQ was exceeded for lead in less than 50% of L. rosae analysed, and more than 50% exceeded that for antimony. The weekly consumption of 150?g of L. rosae muscle tissue from these impoundments may pose an unacceptable health risk to rural communities.     

Biochemical and stress responses of rohu Labeo rohita and mrigal Cirrhinus mrigala in relation to acclimation temperatures

The biochemical and stress responses of two Indian major carps, rohu Labeo rohita and mrigal Cirrhinus mrigala were studied after acclimating them to four preset temperatures (26, 31, 33 and 36° C) for 30 days. The blood glucose and liver glycogen levels showed an inverse trend in both the species and were significantly different in L. rohita at higher temperatures. The decrease in the liver glycogen level of C. mrigala , however, was not significant. Plasma cortisol levels increased significantly whereas the ascorbic acid content in the brain and kidney of both the species decreased significantly with increasing temperatures. Total lipid content in the liver of both the species decreased significantly with increasing acclimation temperatures. The phospholipid concentration decreased in L. rohita with increasing acclimation temperatures, and in C. mrigala the values decreased up to 33° C and increased at 36° C. In C. mrigala , the cholesterol level decreased up to 33° C and then increased at 36° C, but the absolute value was lower in comparison to L. rohita . The cholesterol levels, however, were not significantly different in L. rohita . Triglycerides and free fatty acids concentrations decreased significantly with increasing acclimation temperatures in both the species. The present study indicates species-specific metabolic responses of L. rohita and C. mrigala to thermal acclimation.     

The haematology of Rohu, Labeo rohita

Haematocrit values, haemoglobin concentration, erythrocyte counts, leucocyte counts, erythrocyte sedimentation rate and clotting time of Labeo rohita were determined. The variations in haematological parameters in relation to the size and weight of the fish were also examined. Males had higher haematological values than the females. Seasonal and variations due to maturation of gonads were also apparent.     

Microsatellite DNA markers for population-genetic studies of Labeo dyocheilus (McClelland, 1839)

Fifty‐four primers published for six cyprinid fishes were tested to amplify homologous microsatellite loci in Labeo dyocheilus. Fifteen primers yielded successful amplification and seven were polymorphic with 3–9 alleles. To evaluate utility of the identified loci in population genetic study, 84 samples were analysed. The samples were collected from four rivers viz. Beas, Satluj, Yamuna and Jiabharali. The four sample sets displayed different diversity levels, with observed heterozygosity from 0.34 to 0.53. Significant genotype heterogeneity (P < 0.001) over all loci indicated that the samples are not drawn from the same genepool. The identified microsatellite loci are promising for use in fine‐scale population structure analysis of L. dyocheilus.     

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.     

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.     

Assessing Genetic Differentiation in Geographic Populations of Labeo calbasu Using Allozyme Markers

The population structure of Labeo calbasu from 11 rivers belonging to the Indus, Ganges, Bhima, Mahanadi, and Godavari basins was investigated using allozyme marker systems. Seven out of 20 allozyme loci (35%) were polymorphic (P < 0.99). Both probability and score tests indicated significant deviation of genotype proportions from Hardy–Weinberg expectations at two loci, XDH* (Mahanadi, Bhima, and Godavari) and G6PDH* (Mahanadi). A pairwise genetic homogeneity test and F _ST values indicated a low-to-moderate level (0.0515) of genetic structuring in the wild population of L. calbasu. AMOVA analysis also indicated moderate differentiation among the samples from different river basins. Analysis for genetic bottleneck was performed under the infinite allele model. The study revealed nine genetic stocks of L. calbasu from the natural population across Indian rivers. Evidence of genetic bottlenecks in some rivers was also revealed.     

Effect of Mangifera indica kernel as a feed additive on immunity and resistance to Aeromonas hydrophila in Labeo rohita fingerlings

The study evaluated the efficacy of dietary doses of Mangifera indica (mango) kernel on the immune response and disease resistance of Labeo rohita fingerlings against the bacterial pathogen Aeromonas hydrophila infections in. L. rohita fingerlings fed diet containing 0 (Control), 1g, 5 g, 10 g mango kernel kg(-1) dry diet for 60 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio, blood glucose), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined at 20, 40 and 60 days of feeding. Fish were challenged with A. hydrophila 60 days post feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with mango kernel showed enhanced superoxide anion production, lysozyme, serum bactericidal, serum protein, albumin (P<0.05) compared with the control group. The mortality (%) was recorded up to 10 th day post-challenge. Less survivability was observed in control group (50%) up to day 10 after infection. The survivability was higher in experimental diets. The group fed 5 g kernel kg(-1) dry diet showed highest percentage survival (98%). These results indicate that mango kernel stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.     

Isolation and characterization of hypoxia inducible heme oxygenase 1 (HMOX1) gene in Labeo rohita

The HMOX1 gene plays role in several biological processes and is also responsive to hypoxia stress. Freshwater carp fish, Labeo rohita, is reported as hypoxia sensitive, but the information of annotated hypoxia genes in public domain is very scanty for this species. Here, an attempt was made to isolate and characterize HMOX1 gene in L. rohita using information from zebrafish. HMOX1 gene was obtained by mapping HMOX1 protein of zebrafish over assembled genome of L. rohita. Aligned region was used for designing primers for HMOX1 amplification. Eight overlapping sets of primers were designed for amplifying ~540 bp long successive overlapping fragments. Splicing of overlapping amplicons generated 3715 bp fragment that was confirmed as HMOX1 gene having full coding region with 6 exons between 184 and 2156 bp positions. HMOX1 characterization is an initiative for L. rohita genes annotation to support the characterization of new genes in the important species.     

The haemoglobins of Labeo umbratus (Smith) and Labeo capensis (Smith)

The haemoglobins of both L. umbratus and L. capensis separated into four components on polyacrylamide electrophoresis. The isoelectric points of these fractions were found to be 9.35, 6.89, 6.63 and 6.40. Organic nucleotide triphosphate concentration in the blood of L. capensis was found to be 56.72 ± 18.01 mg% and 58.72 ± 17.9 mg% in the case of L. umbratus . It is not possible to distinguish between L. umbratus and L. capensis purely on the basis of the haemoglobin electropherogram.     

Perturbations in nitrogen metabolic profiles in the tissues of fish, Labeo rohita exposed to fenvalerate.

Changes in the levels of nitrogen metabolic profiles in gill, brain, liver and muscle tissues of fish, Labeo rohita exposed to sublethal concentration (10 micrograms/L) of fenvalerate for 6, 12, 24 and 48 hours were studied. Ammonia content was decreased, whereas urea and glutamine levels were increased in the tissues of fenvalerate exposed fish. Changes were more pronounced with an increase in the period of exposure. Within the tissues, the alterations in excretory products were in the following order: liver greater than muscle greater than brain greater than gill. The survivability of fish in the polluted habitats might be due to the operation of compensatory mechanism in the metabolic profiles such as detoxification and transamination of more toxic ammonia by conversion to less toxic compounds like urea and glutamine.     

Effect of Achyranthes aspera on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila

Achyranthes aspera seed was incorporated in the diets (at 0.01%, 0.1% and 0.5%) of Labeo rohita, rohu fingerlings (3.0+/-0.4 g). After 2 weeks, the fish were immunized with heat-killed Aeromonas hydrophila, and after a further 2 weeks the rohu were experimentally infected with Aeromonas hydrophila (ATCC 49140). After 7 days blood and serum were sampled to determine superoxide anion production, bactericidal activity, lysozyme, serum protein, albumin, globulin, serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and alkaline phosphatase (ALP). Superoxide anion production, serum bactericidal activity, lysozyme, ALP, serum protein, albumin:globulin ratio (A/G) were enhanced in Achyranthes treated groups compared to the control group. SGOT and SGPT levels were elevated in control group, but in Achyranthes treated groups the levels were similar to the uninfected-control group. Higher cumulative mortalities were observed in the control group (77%) up to day-9 after infection. This gradually decreased with increasing dose of Achyranthes, 66% mortality in 0.01% group, 57% mortality in 0.1% group and 28% mortality in 0.5% group. These results indicate that Achyranthes aspera stimulates immunity and increases resistance to infection in L. rohita.     

Purification and properties of cytosolic alanine aminotransferase from the liver of two freshwater fish, Clarias batrachus and Labeo rohita

Cytosolic alanine aminotransferase (c-AAT) was purified up to 203- and 120-fold, from the liver of two freshwater teleosts Clarias batrachus (air-breathing, carnivorous) and Labeo rohita (water-breathing, herbivorous), respectively. The enzyme from both fish showed similar elution profiles on a DEAE-Sephacel ion exchange column. SDS-PAGE of purified enzymes revealed two subunits of 54 and 56 kDa, in both fish. The apparent Km values for l-alanine were 18.5+/-0.48 and 23.55+/-0.60 mM, whereas for 2-oxoglutarate the Km values were observed to be 0.29+/-0.023 and 0.33+/-0.028 mM for the enzyme from C. batrachus and L. rohita, respectively. With l-alanine as substrate, aminooxyacetic acid was found to act as a competitive inhibitor with KI values of 6.4 x 10(-4) and 3.4 x 10(-4) mM with c-AAT of C. batrachus and L. rohita, respectively. However, when 2-oxoglutarate was used as substrate, aminooxyacetic acid showed uncompetitive inhibition with similar KI values for purified c-AAT from both fish. Temperature and pH profiles of the enzyme did not show any marked differences between the two fish examined. These results suggest that liver c-AAT, isolated from these two fish species adapted to different modes of life, remain unaltered structurally. However, at the kinetic level, liver c-AAT from C. batrachus exhibits significantly higher affinity for the substrate l-alanine and decreased affinity for its metabolic inhibitor, in comparison to that of the enzyme purified from L. rohita. Such functional changes seem to be of physiological significance and also provide preliminary evidence for subtle changes in the enzyme as a mark of metabolic adaptation in the fish to different physiological demands.