Monoclonal antibody-based immunodot for evaluating serum immunoglobulin levels in rohu (Labeo rohita)

Immune response in rohu treated with an immunomodulator is usually evaluated employing either non-specific immune parameters or traditional antibody-based tools. In the present study, a monoclonal antibody-based immunodot has been developed for evaluating antibody titre in rohu as a preliminary tool to ensure antibody response due to the effect of an immunomodulator, which can be used for routine field level analysis. The immunodot was sensitive enough to determine rohu immunoglobulin up to 15 μg/ml. Application of the immunodot for evaluating enhancement in immune response could be successfully demonstrated in probiotic fed rohu.     

Effect of dietary L-carnitine supplements on growth and body composition of fingerling rohu, Labeo rohita (Hamilton)

Rohu, Labeo rohita (Ham.), fingerlings (average wt 3.38 g) maintained in 25 m² cement tanks were fed fish-meal-based, 300 g kg⁻¹ protein diets containing different levels (0, 0.25, 0.5, 0.75 and 1.0 g kg⁻¹) of L -carnitine in triplicate over a period of 126 days. Fish that received 0.5 g kg⁻¹ carnitine showed significantly higher growth than those fed the basal diet or the rest of the carnitine diets. Fat digestibility, food conversion efficiency and protein efficiency ratio were better in carnitine-fed fish; however, viscerosomatic and hepatosomatic indices decreased. Muscle proximate composition was affected as a result of carnitine treatment. The results of the present study demonstrate a positive effect of carnitine on the growth and body composition of rohu.     

The potential of periphyton-based culture of two Indian major carps, rohu Labeo rohita (Hamilton) and gonia Labeo gonius (Linnaeus)

The effects of periphyton, grown on bamboo substrates, on growth and production of two Indian major carps, rohu, Labeo rohita (Hamilton) and gonia, Labeo gonius (Linnaeus), were studied at the Bangladesh Agricultural University, Mymensingh. For each species, five ponds were provided with bamboo substrates and five ponds without substrate (control). Ponds were stocked at a rate of 10 000 ha^?1 in both treatments. There was no discernible difference in water quality parameters between treatments. A large number of plankton (39 genera) showed periphytic nature and colonized the bamboo substrates. Rohu grew faster, resulting in a 77% higher net production (P < 0.05) in the ponds with bamboo substrates compared with the ponds without substrate. In contrast, the growth and production of gonia did not vary significantly (P > 0.05) between the substrate and control ponds. Rohu seems to be a more suitable candidate for periphyton‐based aquaculture systems than gonia.     

Allozyme variation of hatchery and river populations of rohu (Labeo rohita, Hamilton) in Bangladesh

The genetic variations of rohu (Labeo rohita, Hamilton) sampled from five hatchery populations (Arabpur, Brahmaputra, Comilla, Kishorganj and Natore) and three major river populations (the Halda, the Jamuna and the Padma) were analysed by allozyme electrophoresis. Ten enzymes encoded by 11 loci were screened, and six were polymorphic. Alleles at three loci (Est‐1^*, Gpi‐1^* and Gpi‐2^*) proved variable for hatchery and river populations, and the Mdh‐2^* locus exhibited heterozygous genotypes for river populations only. Polymorphic loci per population (27.3±5.3%), heterozygous loci per individual (15.5±1.2%) and relative gene diversity (0.27±0.08) in river populations were higher than those for hatchery populations (25.5±1.8%, 10.7±1.6% and 0.25±0.01 respectively). Also, the observed heterozygosity (Ho) and expected heterozygosity (He) (0.09±0.03 and 0.14±0.04 respectively) in river populations were higher than those in hatchery populations (0.08±0.01 and 0.11±0.01 respectively). The lower levels of genetic variability in hatchery populations suggested the occurrence of inbreeding and/or genetic drift. The pairwise population differentiation (F_ST) values showed a lower level of genetic differentiation between hatchery and river population pairs. The unweighted pair‐group method with arithmetic mean dendrogram of Nei's genetic distances showed a relationship between the genetic distance and geographic distance. The populations were clustered into three groups: the Padma in one group, the Halda in second group and the Jamuna, including five hatcheries, in the third group. Highly diversified rohu individuals were observed in the Padma and Halda Rivers, whereas less genetically variable individuals were found in the Jamuna River and five hatcheries. These findings can be useful for rohu hatchery propagation to enhance the sustainable aquaculture production.     

Optimum formulation of feed for rohu, Labeo rohita (Hamilton), with biofloc as a component

The present study was conducted to design an optimum feed mix for the growth of rohu (Labeo rohita) in light-limited indoor culture with biofloc as a component along with fish feed. Eighteen 700-L tanks were utilized for producing biofloc using aquaculture effluent from a nearby carp culture pond. Fifteen different feed mixes were prepared using fish feed and biofloc in dry (4 % moisture content) and wet (90 % moisture content) forms at different proportions and used in feeding trials (three replications) conducted in forty-five 50-L glass aquarium stocked with three rohu fingerlings with average individual weight of 20 ± 1.5 g for a period of 90 days. Fish survival was 100 % in all the treatments. Mixture design was used to obtain a solution of best combination of feed source to obtain the optimum growth parameters of rohu. Optimum growth parameters (net yield, specific growth rate, protein efficiency ratio and feed conversion ratio) of rohu were obtained at feed mix containing 50 % fish feed and 50 % wet floc. The nutritional quality of biofloc was found to be quite suitable for rohu. The images of 3-week-old biofloc captured in scanning electron microscope (SEM) indicated the presence of different types of bacteria, algae, protozoa, rotifers, etc. in different sizes ranging from 10 to 100 μm.     

In vitro assay of plant protease inhibitors from four different sources on digestive proteases of rohu, Labeo rohita (Hamilton), fingerlings

The in vitro inhibitory effect of protease inhibitors from four seed extracts (soybean, grasspea, black gram and horse gram) on digestive proteases of rohu was assessed by enzyme inhibition assay and substrate sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. High proteolytic activity was detected in the intestinal extract of rohu (Labeo rohita) fingerlings at two different pH ranges (8–8.5 and 10–11). That protein digestion occurs mainly in the alkaline condition in this fish without a stomach is evident from very high trypsin activity (0.95±0.04 benzoyl‐dl ‐arginine‐p‐nitroanilide U mg protein⁻¹) in the intestine. In case of grass pea seed, more than 50% inhibition of alkaline protease activity was recorded when the ratio of inhibitor to enzyme was 9.41 μg U⁻¹. More than 40% inhibition of protease activity was recorded in case of horse gram seed when the ratio of inhibitor to enzyme was 5.51 μg U⁻¹. Black gram at 11.0 μg U⁻¹ and soybean seed proteins at 62.75 μg U⁻¹ resulted in 50% and more than 30% inhibition of digestive protease activity in rohu fingerlings respectively. A plot of the inhibition values obtained by changing the relative concentrations of enzyme/inhibitor resulted in different dose–response curves for different protein sources. The use of substrate gel electrophoresis allowed the visualization of the aforementioned differences in inhibition. Each seed extract produced a characteristic profile of protease inhibition. It is concluded that protease inhibitors present in plant protein sources adversely affect the digestive proteases in fish and hence there is a need to eliminate/reduce the amount of such inhibitors through proper processing before incorporation into aquafeeds.     

Effect of fermentation on the nutritive value of sesame seed meal in the diets for rohu, Labeo rohita (Hamilton), fingerlings

Six isonitrogenous (350 g kg⁻¹ crude protein) and isoenergetic (17573 kJ kg⁻¹) experimental diets incorporating raw and fermented sesame ( Seasamum indicum ) seed meal at 200, 300, and 400 g kg⁻¹ into a fishmeal based diet were fed to rohu Labeo rohita fingerlings for 60 days and the growth performance and feed utilization efficiency of the fish was studied. The antinutritional factor phytic acid, from raw sesame seed meal, could be reduced below detection limit by fermentation with lactic acid bacteria ( Lactobacillus acidophilus ). Fermentation of the oilseed meal resulted in reduction of the tannin content from 20 to 10 g kg⁻¹. In terms of growth response, feed conversion ratio and protein efficiency ratio, a diet containing 400 g kg⁻¹ fermented sesame seed meal resulted in a significantly ( P  < 0.01) best fish performance. In general, growth and feed utilization efficiencies of fish fed fermented sesame seed meal diets were superior to those fed raw oilseed meal diets. Apparent protein digestibility (APD) values decreased with increasing levels of raw oilseed meal. APD was, however, significantly ( P  < 0.01) higher at all levels of incorporation of fermented sesame seed meal, while diets containing raw oilseed meal resulted in poor protein and lipid digestibility. Carcass protein and lipid contents of fish fed fermented sesame seed meal diets increased with increasing level of incorporation, being highest with 400 g kg⁻¹ fermented oilseed meal-containing diet. The results showed that sesame seed meal may be incorporated in carp diets up to 200 g kg⁻¹ and 400 g kg⁻¹ in raw and treated (fermented) forms respectively.     

Modulation of the innate immune response of rohu Labeo rohita (Hamilton) by experimental freshwater lice Argulus siamensis (Wilson) infection

The study was undertaken to determine the modulation in innate immune response of rohu (Labeo rohita) during experimental freshwater lice Argulus siamensis infection. Results showed that serum α‐2 macroglobulin (α‐2M) activity, ceruloplasmin level and alternative complement activity were significantly (P<0.05) lower in fish at different degrees of lice infection in comparison with uninfected control. No significant difference (P>0.05) in haemagglutination titre was observed in fish with low‐ and high‐degree lice infections as compared with uninfected control. The serum lysozyme level was significantly (P<0.05) lower in low degree of lice infection as compared with control fish. The total serum antiprotease, myeloperoxidase activity and total protein level were not significantly different (P>0.05) in different degrees of lice‐infected fish with respect to the control fish. The study indicated that A. siamensis infection modulated the immune system of rohu by suppressing the α‐2M, serum complement activities and ceruloplasmin level and through induction of stress response. The baseline data obtained in the present study have tremendous importance in understanding the susceptibility of rohu to different degrees of parasitosis and might be useful in controlling this dreaded ectoparasitic infection in fish.     

Differential inhibition of digestive proteases by tannin in two size groups of rohu (Labeo rohita,Hamilton): A biochemical and zymography study

This study was intended to evaluate tannin mediated inhibition of digestive proteases in two different size groups (fingerlings, F, 4.5 ± 0.7 cm; advanced fingerlings, AF, 18.2 ± 1.6 cm) of rohu, Labeo rohita. Graded levels (50, 100, 150 and 200 nM) of tannin (Gallotannin, 99% purity) were added to the enzyme extracts (30°C, 1 hr) prior to determination of enzyme activities. Changes in the activity of trypsin, chymotrypsin and total protease in relation to the control sets were determined through biochemical assay of enzymes and SDS‐PAGE zymography. The study revealed that tannin significantly inhibited trypsin, chymotrypsin and total protease activities in a dose dependent manner as evident from the regression equations. The degree of the inhibition appeared to be significantly higher for the F in contrast to the AF (F_5,66 = 282.311; p < 0.0001). Trypsin activities were reduced by 8.04 ± 0.19% to 52.68 ± 0.72% and 5.61 ± 0.22% to 39.46 ± 0.19% in F and AF, respectively. The reduction in Chymotrypsin activities ranged between 16.11 ± 0.03% to 38.02 ± 0.27% in F and 6.31 ± 0.07% to 22.80 ± 0.32% in AF. Total protease activities were reduced by 10.9 ± 0.07% to 49.60 ± 0.32% in L. rohitaF, whereas, it ranged between 5.19 ± 0.06% to 32.60 ± 0.13% in AF. On a comparative scale, the difference in tannin induced inhibition in F and AF were more prominent for trypsin and chymotrypsin than total protease. Further, nine protease activity bands (15.9–69 kD) with different electromobility were noticed in both the size groups. Subsequent densitometry analysis revealed that the average densities of the protease activity bands were gradually decreased with increasing level of tannin exposure (50–200 nM). The study might indicate adaptive tolerance to tannin in larger size groups and emphasizes the need for removal of tannin in the plant feedstuffs, especially for feeding the fingerlings of L. rohita.     

Effect of dietary Curcuma longa on enzymatic and immunological profiles of rohu,Labeo rohita (Ham.), infected with Aeromonas hydrophila

The aim of this study was to evaluate the dietary dosages of turmeric that enhance immune response and disease resistance against the opportunistic pathogens Aeromonas hydrophila in Labeo rohita fingerlings. Hence, four different dosages of turmeric at 0.1, 0.5, 1.0 and 5.0 g kg^?1 feed were given to the fingerlings of L. rohita for 60 days at 4% body weight. At every 20‐day interval, different biochemical, haematological, enzymatic and immunological parameters of fish were evaluated. After 60 days, fish were challenged with A. hydrophila. The mortality (%) was recorded on the tenth day post challenge. Most of the immune parameters including lysozyme activity, superoxide anion production and serum bactericidal activity were significantly (P<0.05) higher on 60 days of feeding of 1.0 g of turmeric per kg of feed. Challenge study indicated 100% and 89% survivability in the group of fish fed with 5.0 and 1.0 g of turmeric per kg of feed respectively. Feeding of turmeric might have maintained long‐term protection in fish by elevating the nonspecific immune system such as Nitroblue tetrazolium (NBT), lysozyme and serum bactericidal activity. The result showed that turmeric at a dose of 1.0 g kg^?1 feed for 60 days provided the greatest protection to pathogen challenge.     

Stress mitigating and growth enhancing effect of dietary tryptophan in rohu (Labeo rohita,Hamilton, 1822) fingerlings

An experiment was conducted to study the stress mitigation and growth enhancing role of dietary l-tryptophan (TRP) under thermal stress in rohu, Labeo rohita fingerlings for 45 days. Seven hundred and twenty fishes were distributed in three major groups that are ambient temperature (26 °C), 34 and 38 °C in triplicate following a complete randomized design. Acclimation of fishes to 34 and 38 °C over average ambient temperatures were carried out at 1 °C/day. Each group was fed with a diet supplemented with 0, 0.36, 0.72 or 1.42 % l-TRP. Results showed that blood glucose and serum cortisol level were found to be significantly higher (p < 0.05) in the higher temperature groups than the ambient temperature group. Similarly, aminotransferase, lactate dehydrogenase, malate dehydrogenase, CAT, superoxide dismutase activities were found to be significantly higher (p < 0.05) in the control groups (0 % l-TRP) and decreasing activities of these enzymes were observed with the increasing level of dietary l-TRP. In different temperature groups, l-TRP-supplemented groups were found to have higher (p < 0.05) growth, RGR and PER. The results obtained in the present study indicate that dietary l-TRP mitigates thermal stress and enhances growth. From the present study, we can conclude that dietary supplementation of l-TRP at the 0.72 % level in the diet is found to be optimum to reduce thermal stress even up to 38 °C in rohu, L. rohita. The baseline data obtained here could be useful for the farmers to formulate feeds to culture the fish in different agro-climatic zones.     

Evaluation of selected metal nanoparticles on hatching and survival of larvae and fry of Indian major carp,rohu (Labeo rohita)

Different laboratory synthesized metal nanoparticles viz. Copper oxide (CuO), Zinc oxide (ZnO) and silver doped titanium dioxide (Ag‐TiO₂) were studied for their effect on hatching and survival of larvae and fry of Indian major carp, rohu, Labeo rohita both in direct application in tank water & coated onto tanks. Among these nanoparticles, CuO and ZnO nanoparticles exhibited highest percentage of hatching in both direct addition (78.0 ± 3.1% and 78.05 ± 4.2%, respectively) and coating onto tanks (58.6 ± 2.1% and 61.2 ± 2.7%, respectively) at 1 mg mL^?1 while least percentage of hatching was recorded in Ag‐TiO₂ nanoparticles irrespective of its concentration & mode of supplementation. Highest survival of L. rohita fry (50.13 ± 2.2%) was observed after 15 days post hatching in CuO coated tanks followed by ZnO coated tanks (38.6 ± 2.8%) while least was recorded in Ag‐TiO₂ coated tanks (22.53 ± 3.0%). However in control tanks coated with Poly‐Urethane base with hardener and uncoated control tanks, the survival was 42.4 ± 1.2% and 41.36 ± 1.8% respectively. Further, significantly lower microbial load of water was recorded in CuO nanoparticles coated tanks (1.5 × 10¹⁰ CFU L^?1) as compared to uncoated control tanks (1.1 × 10¹⁶ CFU L^?1) without affecting water quality parameters. On the other hand, in Ag‐TiO₂ coated tanks, significantly lower microbial load (1.0 × 10⁶ CFU L^?1) as compared to uncoated control tanks at 15 days post hatching was recorded. However, Ag‐TiO₂ was toxic to L. rohita larvae & fry both in direct application and coating onto tanks. Considering the beneficial effects of CuO nanoparticle application, it has the scope of being used in a more eco‐friendly way in hatchery operations.     

Ecological intensification for feeding rohu Labeo rohita (Hamilton, 1822): A review and proposed steps towards an efficient resource fishery

Labeo rohita (rohu) is a commercially popular Indian major carp species in polyculture fish farming system. It feeds mainly on plankton. The adults of rohu have a strong positive selection for phytoplankton, whereas the fingerlings have a strong positive selection for both zooplanktonic and smaller phytoplanktonic sources. Conversely, adults of rohu exhibit a strong negative selection for all zooplanktonic sources and a positive selection for most phytoplanktonic sources. In periphytic environments, the fish grow better feeding on the sub‐periphytic zone in proximity to the substrate. We conclude that rohu is an excellent aquaculture candidate, thanks to its ability to explore natural resources (plankton and periphyton) and transform them to fish biomass in polyculture practice. In contrast to over‐reliance on artificial feed, translation of natural resources for exploration by rohu could be the next step for sustainable carp‐based polyculture practice.     

The apparent total and nutrient digestibility of sal seed (Shorea robusta) meal in rohu, Labeo rohita (Hamilton), fingerlings

The apparent digestibility of both raw and treated (16 h soaked in water) deoiled salseed (Shorea robusta) meal by Labeo rohita (Hamilton) fingerlings was evaluated at 20, 30, 40, 50 and 60% levels of incorporation for each, using fish meal based feed as the reference diet. The tannin content from salseed meal could be reduced from 3.4% (in raw salseed meal) to 1.1, 0.9, 0.8 and 0.7% by soaking in water at room temperature for 30 min, 1 h, 2 h and 16 h, respectively. The digestibility of dry matter, protein, lipid and ash declined with increasing levels of inclusion of raw and soaked salseed meal. The diets containing soaked salseed meal resulted in better digestibility of nutrients at all levels of inclusion as compared to those obtained with diets that incorporate raw salseed meal. The results indicate that deoiled salseed meal can be incorporated upto a level of 30% in processed (16 h soaking in water) condition by reducing tannin.     

Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton)

Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita , and the brain of catla, Catla catla , respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 °C with an optimum of 28 °C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 °C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp . were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.     

Characteristics of Collagen from Rohu (Labeo rohita) Skin

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from rohu skin with the yield of 64.2 and 6.8% (dry weight basis), respectively. Both collagens had glycine as the major amino acid with imino acid content of 196–202 residues/1,000 residues and were characterized as type I collagen with molecular composition of (α1)₂α2-heterotrimer. Fourier transform infrared spectra of both collagens were similar, with no shift in wavenumber of all amide bands. The T_max value of ASC and PSC was 36.40 and 35.48°C, respectively. The zero surface net charge of ASC and PSC was found at pH 5.9 and 5.3, respectively.