Immunomodulatory effect of recombinant RNA‐dependent RNA polymerase protein of Macrobrachium rosenbergii nodavirus in giant freshwater prawn M. rosenbergii

The present study evaluated the role of recombinant RNA‐dependent RNA polymerase (RdRp) protein of Macrobrachium rosenbergii nodavirus (MrNV) in modulating the immune response and in reducing MrNV load in infected prawn. In the first experiment, prawns (25–30 g) were injected with recombinant RdRp protein (RP) at a concentration of 0, 1.0 and 10 μg, and immune parameters and expression of some immune‐related genes were measured up to 14 days post injection (p.i.). In the second experiment, early juveniles were injected with a similar dose of RdRp and animals were challenged by immersion with MrNV. The infection status was detected in muscles by nested RT‐PCR up to 21 days post challenge. Prawn injected with higher concentration of RP showed significantly higher total haemocyte count at different period post injection. Significant up‐regulation of immune‐related genes was observed within 24 h in prawn treated with lower dose of RP and after 7 days p.i. at higher level of RP injection compared with adult control. Most of the tested samples (63%) were found to be RT‐PCR positive for MrNV at 48 h of post‐immersion challenge. After 14 days, MrNV was detected only in control prawn, while both RP‐injected groups were MrNV negative. This study elucidated the potential viral load reduction role played by RdRP in MrNV‐infected prawn.     

Growth, reproductive performance, muscle and egg composition in grass carp, Ctenopharyngodon idella (Valenciennes), fed hydrilla or formulated diets with varying protein levels

Growth, reproductive performance, muscle and egg composition were investigated in grass carp, Ctenopharyngodon idella (Valenciennnes), fed hydrilla or formulated diets with varying protein levels. Five experimental diets, with varying levels (20%, 25%, 30%, 35% and 40%) of crude protein (CP), were used. One of the fish groups was fed hydrilla. Fish (44.1±0.3 cm; 913±9 g) were stocked (20 tank^?1) in outdoor concrete tanks (20 × 10 × 1.5 m) in duplicate, and fed to satiation, twice daily, at 09:00 and 17:00 hours for the experimental duration of 360 days. High (P<0.05) weight gain was recorded in fish fed 30% and 35% CP diets. However, values for gonadosomatic index (GSI), egg diameter, relative fecundity (eggs kg^?1 body weight), fertilizability and hatchability (%) were comparable (P>0.05) in fish at ≥25% of dietary protein intake. Hydrilla‐fed fish exhibited lower (P<0.05) values for the measured parameters. Crude protein content in muscle increased with dietary protein level. Highest (P<0.05) muscle protein was obtained in fish fed 35% CP diet. Muscle fat was comparable (P>0.05) among fish receiving formulated diets. Ash content was not significantly (P>0.05) different among fish of different dietary groups. Moisture content in fish fed formulated diets, with the exception of 20% CP diet, did not vary significantly (P>0.05). Eggs of fish fed formulated diets contained higher CP and fat contents than those of hydrilla‐fed fish. High (P<0.05) moisture content was noted in the eggs of hydrilla‐fed fish. Ctenopharyngodon idella fed formulated diet, with a minimum of 25% CP, showed better reproductive performance than those fed hydrilla.     

Impact of Aeration on the Growth Performance of Silver Barb,Puntius gonionotus,during Fingerling Rearing

Impact of aeration on growth of silver barb, Puntius gonionotus during fingerling rearing was studied through a 100‐d rearing experiment conducted in 18 concrete tanks of 50 m² (10 × 5 × 1.2 m) size. Fry (0.74 ± 0.27 g, 35 ± 6 mm) were stocked in the experimental tanks at three stocking densities (25, 50, and 75 fry/m²) and were evaluated with and without provision of 6 h (2400–0600 h) of night time aeration. Aeration resulted in higher pH and dissolved oxygen regime and increased fingerling length and weight. The results suggest a rearing density of 75/m² to be ideal for rearing fry to fingerling of this species when aeration is provided, whereas, under non‐aerated condition, rearing the fry to fingerling stage at 50/m² was found advantageous over those at 25 and 75/m².     

Effect of brewer’s yeast on immune response of giant freshwater prawn, Macrobrachium rosenbergii, and its resistance to white muscle disease

The present study was conducted to find the effect of dietary brewer’s yeast on growth, survival, immune response, and resistance to white muscle disease in giant freshwater prawn, Macrobrachium rosenbergii. The brewer’s yeast was supplemented at graded levels 0, 5, 10, and 20?g per kg diet, and the experiment was conducted for 75?days. After the feeding trials, growth, survival, immune parameters like prophenoloxidase activity, respiratory burst, and total hemocyte count were assessed. The growth, specific growth rate, and survival were not found significantly different among the treatment and control groups “prophenoloxidase activity and respiratory burst” were found significantly different (p?<?0.05) among the treatment and control groups. The total hemocyte count was significantly different (p?<?0.05) among control and treatment groups. Cumulative percent survival was higher in M rosenbergii fed 1?% brewer’s yeast-supplemented diet when they were challenged with white muscle disease virus (MrNV (Macrobrachium rosenbergii nodavirus) and XSV (extra small virus)). The present work revealed that the incorporation of brewer’s yeast in the diet is effective in enhancing immune response and controlling the white muscle disease in M rosenbergii.     

Effect of a Previous Washing Step with Tannic Acid on the Quality of Minced Horse Mackerel (Trachurus trachurus) During Frozen Storage

Fatty fish have been recognized as potential raw material for production of minced meat; however, they are prone to oxidation and further deterioration. In the present study, the effect of washing and antioxidant (tannic acid) treatment on the quality of minced meat of Trachurus trachurus (horse mackerel) during frozen storage was observed. Minced meat of Trachurus trachurus was divided into three lots (T0, T1, and T2). T1 was washed with cold water, T2 with cold water containing tannic acid (100 mg/kg), and T0 was not washed. All the lots were frozen at ?40°C and stored at ?20 ± 2°C for 125 days and were subjected to biochemical, microbiological, and sensory evaluation at regular intervals of 25 days. The antioxidant treatment with tannic acid at the dosage used was found effective in minimizing the rancidity problems of minced meat (T2), compared to T0 and T1. During the whole period of storage, samples from T2 showed good quality in terms of microbiological, biochemical, and sensory analysis compared to T1 and T0.     

Purification and characterization of a β–glucan binding protein from the haemolymph of freshwater prawn Macrobrachium rosenbergii

β‐glucan binding protein (βGBP), a pattern recognition protein was purified from the haemolymph of freshwater prawn Macrobrachium rosenbergii by heparin affinity chromatography that showed a single band in native gradient PAGE. The β‐glucan binding property of the purified protein was confirmed in a phenoloxidase (PO) assay, where addition of βGBP along with β‐glucan increased the specific PO activity compared with that of β‐glucan alone. The molecular weight of the βGBP was found to be ~316 kDa on gel filtration chromatography. In SDS‐PAGE, βGBP molecule was reduced to one polypeptide chain of molecular weight ~113 kDa. Thus the βGBP in M. rosenbergii is possibly a homotrimeric molecule. The purified sample run on unreduced condition in SDS‐PAGE also revealed a similar size band (~113 kDa) and hence, the polypeptide chains of βGBP are held by non‐covalent interactions. The purified βGBP samples run in native PAGE was stained positively with alcian blue for carbohydrates and Sudan black for lipids indicating the βGBP to be a glycolipoprotein. With rabbit polyclonal anti‐βGBP serum developed, an indirect ELISA was standardized and the normal βGBP concentration in adult M. rosenbergii serum was quantified to be ~2 mg mL^?1. Furthermore, the applicability of the developed ELISA is discussed.     

Efficacy of 17 α‐methyl testosterone and letrozole on sex reversal of protogynous grouper,Epinephelus tauvina (Forskal, 1775) and spawning performance of sex‐reversed males

This study aimed to test the efficacy of 17 α‐methyl testosterone (17 α‐MT) alone and in combination with letrozole, an aromatase inhibitor, for the induction of sex reversal in protogynous greasy grouper, Epinephelus tauvina. Further, the long‐lasting effects of these treatments and spawning performance of sex‐reversed males were also investigated. Greasy grouper with oocytes in the perinucleolus stage were implanted with 5 mg 17 α‐MT kg^?1 body weight (T₁), 5 mg 17 α‐MT and 0.2 mg letrozole kg^?1 body weight (T₂) and 5 mg 17 α‐MT with 0.4 mg letrozole kg^?1 body weight (T₃) and no androgens/enzyme inhibitor implanted (C). The 17 α‐MT alone and in combination of letrozole‐induced sex reversal in greasy grouper, whereas untreated control fish (C) showed normal ovarian development. However, T₂ and T₃ group showed 100% sex reversal and completion of spermatogenesis up to functional male phase in 2 and 3 months, respectively, whereas T₁ group resulted in only 66.67% functional male with motile spermatozoa after 4 months. Sex‐reversed males successfully fertilized the eggs during induced spawning. There were significant differences on fertilization and hatching rates between T₂ group (79.00 ± 4.36%; 77.67 ± 2.87%, respectively) and T₁ group (57.67 ± 3.17%; 63.87 ± 2.91%, respectively). The result suggested that 17 α‐MT (5.0 mg kg^?1 BW) in combination with letrozole (0.2 mg kg^?1 BW) has the potential to produce 100% sex‐reversed male in short period in greasy grouper, which might greatly help in seed production of greasy grouper.