Development,distribution and expression of a DNA vaccine against nodavirus in Asian Seabass,Lates calcarifier (Bloch, 1790)

Fish nodavirus (betanodavirus), a viral pathogen responsible for viral nervous necrosis (VNN) was isolated from infected Asian sea bass (Lates calcarifer). The distribution, clearance and expression of nodavirus vaccine, on the basis of DNA vaccine (pFNCPE42 DNA‐pcDNA3.1) construction, were analysed in tissues of the Asian seabass by PCR, RT‐PCR, ELISA and Immunohistochemistry. Fish immunized with a single intramuscular injection of 20 μg of the pFNCPE42‐DNA vaccine showed a significant increase in the serum antibody level in the 3rd week after vaccination, compared to control eukaryotic expression vector pcDNA3.1 vaccinated fish. Results from PCR studies indicated that the vaccine‐containing plasmids were distributed in heart, intestine, gill, muscle and liver 10 days after vaccination. Clearance of pFNCPE42‐DNA vaccine was studied at 10, 25, 50, 75 and 100 days of post vaccination (d p.v). At 100 days p.v. pFNCPE42‐DNA was cleared from muscle of vaccinated sea bass. In vitro and in vivo expression of fish nodavirus capsid protein gene (FNCP) was determined by fluorescent microscopy. Asian seabass was immunized with pFNCPE42‐DNA vaccine at a dose of 20 μg per fish and were challenged with betanodavirus by intramuscular injection. The vaccinated seabass was protected from nodaviral infection and 77.33% of relative percent survival (RPS) was recorded.     

High efficacy of white spot syndrome virus replication in tissues of freshwater rice‐field crab,Paratelphusa hydrodomous (Herbst)

An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV) of shrimp in different organs of freshwater rice‐field crab, Paratelphusa hydrodomous (Herbst), using bioassay, PCR, RT‐PCR, ELISA, Western blot and real‐time PCR analyses, and also to use this crab instead of penaeid shrimp for the large‐scale production of WSSV. This crab was found to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT‐PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real‐time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production.     

The induction of Hsp70 synthesis by non‐lethal heat shock confers thermotolerance and resistance to lethal ammonia stress in the common carp,Cyprinus carpio (Linn)

Exposure to heat‐shock protein (Hsp) stimulating factors induces Hsp accumulation and confers tolerance to lethal ammonia stress on the common carp Cyprinus carpio. This study investigated whether a non‐lethal heat shock bestowed similar protective effects against ammonia and induced thermotolerance, both thought to be rendered by increased amounts of Hsps. The 30‐min lethal temperature (30 min LHT) and 1‐h lethal ammonia concentration (1 h LCT) for this species occurred at 41°C and 14.2 mg/L NH₃ respectively. Heating juvenile carp (5 cm) from 28°C to 32, 34 and 38°C, with a subsequent 8‐h recovery period augmented tolerance to lethal heat and ammonia perturbation by two to threefold as compared with animals held at 28°C. Protection occurred in conjunction with Hsp70 accumulation in gills, substantiating the role of this Hsp in enhancing the stress tolerance of common carp.     

The potential of mixed culture of genetically improved farmed tilapia (Oreochromis niloticus) and freshwater giant prawn (Macrobrachium rosenbergii) in periphyton-based systems

The production performance of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) and freshwater prawn (Macrobrachium rosenbergii) in periphyton‐based systems were studied in farmers' ponds at Mymensingh, Bangladesh. Fifteen ponds (200–300 m² area and 1.0–1.5 m in depth) were used to compare five stocking ratios in triplicate: 100% GIFT, 75% GIFT plus 25% prawn, 50% GIFT plus 50% prawn, 25% GIFT plus 75% prawn and 100% prawn. Ponds were stocked at a total density of 20 000 GIFT and/or prawn ha^?1. Bamboo poles (mean diameter 6.2 cm and 5.5 pole m^?2) were posted in pond bottoms vertically as periphyton substrate. Periphyton biomass in terms of dry matter (DM), ash‐free DM and chlorophyll a were significantly higher in ponds stocked with prawn alone than in ponds with different combinations of GIFT and prawn. Survival of GIFT was significantly lower in ponds stocked with 100% GIFT (monoculture) whereas, that of prawn was significantly higher in its monoculture ponds indicating detrimental effects of GIFT on prawn's survival. Individual weight gains for both species were significantly higher in polyculture than in monoculture. The highest total fish and prawn yield (1623 kg GIFT and 30 kg prawn ha^?1) over 125–140 days culture period was recorded in ponds with 75% GIFT and 25% prawn followed by 100% GIFT alone (1549 kg ha^?1), 50% GIFT plus 50% prawn (1114 kg GIFT and 68 kg prawn ha^?1), 25% GIFT plus 75% prawn (574 kg GIFT and 129 kg prawn ha^?1) and 100% prawn alone (157 kg ha^?1). This combination also gave the highest economic return. Therefore, a stocking ratio of 75% GIFT plus 25% prawn at a total density of 20 000 ha^?1 appeared to be the best stocking ratio in terms of fish production as well as economics for a periphyton‐based polyculture system.     

Influence of Zn nutrition on the productivity,grain quality and grain biofortification of wheat under conventional and conservation rice–wheat cropping systems

ABSTRACT

This study was conducted to evaluate influence of zinc (Zn) application on productivity, grain biofortification and grain quality of wheat planted under plough tillage (PT) and zero tillage (ZT) systems. Zn was delivered as soil application (10 kg ha^?1), foliage spray (0.025 M) and seed priming (0.5 M) in wheat planted under PT and ZT systems. ZT had higher total soil porosity, total soil organic matter, soil organic carbon and soil microbial biomass carbon than PT. Zn application, by either method, improved grain yield, grain Zn and grain quality in both tillage systems. The grain Zn concentration was 72% and 59% higher with soil-applied Zn in ZT wheat during 2016–2017 and 2017–2018, respectively, compared with no Zn. However, Zn seed priming was the most effective in improving wheat grain yield in both tillage systems. Foliage and Zn soil application were better in improving the indices of Zn use efficiency of Zn. In conclusion, Zn seed priming was the most effective method in improving the wheat grain yield, whereas soil Zn application in ZT and foliar applications in PT were the most effective for grain Zn biofortification. However, Zn soil application was the most cost-effective method of Zn application.     

Soil Application of Boron Improves the Tillering,Leaf Elongation,Panicle Fertility,Yield and its Grain Enrichment in Fine-Grain Aromatic Rice

Boron (B) is one of the essential micronutrients having a specific role, particularly during reproductive phase, in rice. In a previous experiment on aerobic rice, panicle sterility was noted as one of the major challenges. This experiment was conducted to evaluate the influence of soil-applied B on tillering, panicle sterility, water relations, and grain enrichment in fine-grain aromatic rice cultivars ‘Super Basmati’ and ‘Shaheen Basmati’. Boron was soil applied at 0.50, 0.75, 1, 1.25, and 1.50 kg ha^?1 while the control treatment did not receive B. Rate of leaf emergence and elongation and tiller appearance were significantly improved by B application. Likewise, B application also improved the leaf chlorophyll contents and water relations in both rice cultivars. Substantial improvement in kernel yield and yield contributing traits was also observed by B application owing to decrease in panicle sterility. A linear increase in leaf and kernel B contents was observed with increase in B application rate. However, the range for an optimum B application rate is very narrow and increase of B application beyond 1 kg ha^?1 was toxic. In conclusion, soil application of B is an effective way to decrease panicle sterility and increase the kernel yield and grain B enrichment in rice.     

Effect of relaxin on the motility, acrosome reaction and utilization of glucose of fresh and frozen-thawed bovine spermatozoa

The study was conducted to investigate the effect of relaxin on motility, acrosome reaction (AR), viability and utilization of glucose in fresh and frozen‐thawed bovine spermatozoa. Both semen samples were washed twice through centrifugation (5 min at 600 g), and preincubated for 1 h at 39°C for swim up. The swim‐up separated spermatozoa were resuspended in a sperm Tyrode's albumin lactate pyruvate (Sp‐TALP) medium containing 0 (control) and 40 ng/mL porcine relaxin and incubated for 0–6 h. Sperm motility was determined on the basis of movement quality examined by a phase contrast microscope. Sperm viability and AR were evaluated by using the triple staining technique. The incorporation and oxidation of ¹⁴C‐glucose was assessed by a liquid scintillation counter. Motility was improved (P < 0.05) in both fresh and frozen‐thawed spermatozoa by the addition of relaxin to the Sp‐TALP medium, whereas relaxin showed no significant effect on viability in either fresh or frozen‐thawed spermatozoa. The percentage of AR increased (P < 0.05) when fresh or frozen‐thawed spermatozoa were incubated with relaxin. In contrast, the incorporation and oxidation of ¹⁴C‐glucose increased (P < 0.05) in both kinds of spermatozoa incubated with relaxin. Thus the results demonstrated that the addition of relaxin to the Sp‐TALP medium increased the motility, AR and utilization of glucose in fresh and frozen‐thawed bovine spermatozoa.     

Efficacy of two organotin compounds and neem extract against the sorghum shoot fly

The antifeedants triphenyltin acetate and hydroxide and neem seed kernel extracts in two concentrations each, were evaluated as control measures against the sorghum shoot fly,Atherigonu varia soccata Rond. in comparison with the insecticides presently used for this purpose. The antifeedants appeared to be partially effective and were less active than the insecticides.