Development of immunodot blot assay for the detection of white spot syndrome virus infection in shrimps (Penaeus monodon)

The VP 28 gene encoding a structural envelope protein of the white spot syndrome virus (WSSV) was cloned into a pET32a(+) expression vector for the production of the recombinant VP28 protein. A purified recombinant protein of 39.9 kDa size was used for polyclonal antibody production in rabbit. Specific immunoreactivity of the rabbit anti rVP28 antiserum to the viral antigen was confirmed by a Western blot. The specificity of this polyclonal anti‐rVP28 antiserum to detect the presence of the virus in WSSV‐infected Penaeus monodon was verified using a immunodot blot assay. Immunodot blot showed a positive reaction in infected shrimp tissues with prominent colour development using 3,3′,5,5′‐tetramethylbenzidine (TMB) as a chromogenic substrate when compared with 3–3′ diaminobenzidine tetrahydrochloride (DAB). Highest signal intensities of the immunodots were observed in infected shrimp pleopod extracts and haemolymph. On comparison with polymerase chain reaction (PCR), immunodot blot could detect 76% of PCR‐positive WSSV‐infected shrimp samples. Immunodot blot was found to be equivalent to first‐step PCR sensitivity to detect WSSV particles estimated to contain 1.0 × 10⁵ viral DNA copies.     

Effect of Population Levels of Soybean on Physiological Growth Parameters of Sorghum CO 24 in Sole and Intercropped Stand

A field experiment was conducted during kharif, 1985 and summer, 1986 in sandy clay loam soil under irrigated condition at Tamil Nadu Agricultural University, Coimbature, with a view (i) to assess the suitable method of planting of sorghum CO 24 when intercropped with soybean, (ii) to evaluate the suitable planting ratio of sorghum - soybean intercropping system, (iii) to find out the compatible population level of soybean when intercropped with sorghum CO 24.
Direct sown sorghum at 4: 2 and 2: 2 recorded grater leaf area (LA) compared to transplanted sorghum in both kharif and summer. Sorghum transplanted with 14 days old seedlings recorded higher Crop Growth Rate (CGR) compared to 21 days old seedlings. Increased Net Assimilation Rate (NAR) were recorded in sorghum transplanted with 14 days old seedlings as compared to direct sown and 21 days old transplanted crop. Transplanted sorghum recorded higher Relative Growth Rate (RGR) as compared to direct sown sorghum. Summer sorghum recorded higher Relative Growth Rate (RGR) as compared to kharif season.     

Abundance of sulphur‐oxidizing bacteria in coastal aquaculture using soxB gene analyses

Molecular techniques based on sequencing of metagenomic clone libraries provide an insight into the diversity of microbial populations. Using nucleic acid‐based methods, the diversity of soxB genes was examined to detect and characterize sulphur‐oxidizing bacteria in Indian coastal aquaculture environments. Gene‐specific degenerate primers were used to amplify various fragments (710, 753, 483–503, 280 and 239 bp) of soxB genes. Metagenomic clone libraries were constructed for 753, 483–503 and 239 bp fragments of soxB genes. The abundance of soxB revealed the presence of sulphur‐oxidizing organisms. Amino acids in parts of the soxB‐encoded proteins were aligned to known conserved amino acid residues. The level of conservation ranged from 23% to 30%. A phylogenetic tree constructed from aligned amino acid sequences of SoxB revealed different clusters associated with the branches of phototrophic α‐ and γ‐proteobacteria. In general, soxB is widespread among the various phylogenetic groups, although this does not necessarily mean that the organism can use sulphur compounds. Our results suggest that the chemolithoautotrophy based on sulphur oxidation in coastal aquaculture is primarily sustained by the presence of sulphur oxidizers, which involve the soxB gene. This study aids identification of the phylogenetic characteristics related to sulphur bioremediation in poorly characterized coastal aquaculture environments.     

Limited evidence for genetic variation for resistance to the white spot syndrome virus in Indian populations of Penaeus monodon

There has been a highly detrimental impact of the white spot syndrome virus (WSSV) on black tiger shrimp (Penaeus monodon) aquaculture in India. Currently, no cost‐effective measures are available for controlling the disease. One alternative is to improve WSSV resistance through a selective breeding programme for disease‐resistant shrimp, provided that genetic variation exists for this trait. The aim of this study was to evaluate the evidence for genetic variation in resistance to WSSV in P. monodon sourced from Indian populations. Post‐larval shrimp (n=1950) from 54 full‐sibling families were challenged with WSSV using WSSV‐infected mince meat. The heritability was estimated using four different statistical models fitted to the resulting time to death data, including two linear models and two Weibull proportional hazard frailty models. None of the estimated heritabilities were significantly different from zero. We suggest three possible explanations for these results: there actually is very little variation between P. monodon in WSSV resistance and all individuals are highly susceptible to the disease; there is genetic variation in resistance to WSSV in P. monodon but we did not find it in our experiment because the level of challenge in the experiment was too high to allow genetic differences to be expressed; the variation is due to mutations conferring resistance, which are at a low frequency in the population, and we did not sample a broad enough genetic base to capture these mutations.