Biology, Host Range, Pathogenesis and Diagnosis of White spot syndrome virus.

White spot syndrome virus (WSSV) is the most serious viral pathogen of cultured shrimp. It is a highly virulent virus that can spread quickly and can cause up to 100 % mortality in 3-10 days. WSSV is a large enveloped double stranded DNA virus belonging to genus Whispovirus of the virus family Nimaviridae. It has a wide host range among crustaceans and mainly affects commercially cultivated marine shrimp species. The virus infects all age groups causing large scale mortalities and the foci of infection are tissues of ectodermal and mesodermal origin, such as gills, lymphoid organ and cuticular epithelium. The whole genome sequencing of WSSV from China, Thailand and Taiwan have revealed minor genetic differences among different strains. There are varying reports regarding the factors responsible for WSSV virulence which include the differences in variable number of tandem repeats, the genome size and presence or absence of different proteins. Aim of this review is to give current information on the status, host range, pathogenesis and diagnosis of WSSV infection.

White spot syndrome virus: Genotypes, Epidemiology and Evolutionary Studies.

White spot syndrome virus (WSSV) is a pathogen that has emerged globally affecting shrimp populations. Comparison of WSSV genome have shown the virus to share a high genetic similarity except for a few variable genomic loci that has been employed as markers in molecular epidemiology studies for determining the origin, evolution and spread in different geographical regions. Molecular genotyping of WSSV are based on genomic deletions associated with ORF23/24 and ORF14/15 variable regions and the three variable number of tandem repeat regions, ORF75, ORF94 and ORF125. Studies show the prevalence of several genotypes for WSSV with particular genotypes being more prevalent than others in a given geographical area. Deletions associated with ORF23/24 and ORF14/15 variable regions have proven to be of evolutionary significance. Fitness and virulence studies on different genotypes of WSSV suggest that all the strains of WSSV are equally virulent, but the one with smaller genomic size is the fittest. Studies also have shown that mixed genotype infection of WSSV correlates with lower disease outbreaks. This review focuses on the genotyping studies that were undertaken in elucidating WSSV evolution and epidemiology.

Detection of Laem-Singh virus in cultured Penaeus monodon shrimp from several sites in the Indo-Pacific region.

Laem-Singh virus (LSNV) is a positive-sense single-stranded RNA (ssRNA) virus that was recently identified in Penaeus monodon shrimp in Thailand displaying signs of slow growth syndrome. A total of 326 shrimp collected between 1998 and 2007 from countries in the Indo-Pacific region were tested by RT-PCR for evidence of LSNV infection. The samples comprised batches of whole postlarvae, and lymphoid organ, gill, muscle or pleopod tissue of juvenile, subadult and adult shrimp. LSNV was not detected in 96 P. monodon, P. japonicus or P. merguiensis from Australia or 16 P. monodon from Fiji, Philippines, Sri Lanka and Mozambique. There was no evidence of LSNV infection in 73 healthy juvenile P. vannamei collected during 2006 from ponds at 9 locations in Thailand. However, LNSV was detected in each of 6 healthy P. monodon tested from Malaysia and Indonesia, 2 of 6 healthy P. monodon tested from Vietnam and 39 of 40 P. monodon collected from slow-growth ponds in Thailand. A survey of 81 P. monodon collected in 2007 from Andhra Pradesh, India, indicated 56.8% prevalence of LSNV infection but no clear association with disease or slow growth. Phylogenetic analysis of PCR amplicons obtained from samples from India, Vietnam, Malaysia and Thailand indicated that nucleotide sequence variation was very low (>98% identity) and there was no clustering of viruses according to site of isolation or the health status of the shrimp. The data suggests that LSNV exists as a single genetic lineage and occurs commonly in healthy P. monodon in parts of Asia.

Characterization of variable genomic regions of Indian white spot syndrome virus.

A recent study examining genetic variations among the 3 completely sequenced white spot syndrome virus (WSSV) genomes isolated from China, Thailand and Taiwan revealed five major differences among them. Of these differences, a deletion region between ORF 23/24 and a variable region of ORF14/15 prone to recombination were of particular evolutionary significance. Focusing on these regions, 81 WSSV isolates from India were characterized by sequencing polymerase chain reaction (PCR) amplicons. The Indian strains carried a 10,970 bp deletion in the ORF 23/24 region relative to WSSV-TW and WSSV-TH-96-II. Analysis of the ORF 14/15 regions revealed two novel strains of WSSV with unique sequences which could have evolved by recombination. None of the WSSV isolates had a transposase sequence or VP35 gene as reported for Taiwan isolates. The Indian strains were closely related to Thailand strains suggesting movement of a putative ancestor from Thailand to other parts of the world including India.

Rapid detection and enumeration of trh-carrying Vibrio parahaemolyticus with the alkaline phosphatase-labelled oligonucleotide probe.

An alkaline phosphatase (AP)-labelled oligonucleotide probe was developed to detect and enumerate trh(+)Vibrio parahaemolyticus in seafood. The probe was evaluated using 40 isolates of V. parahaemolyticus, 45 isolates of other vibrios and 55 non-vibrio isolates. The probe reacted specifically with V. parahaemolyticus possessing either the trh1 or trh2 variant of the trh gene and was found to be 100% specific for trh(+)V. parahaemolyticus. Using the trh probe, V. parahaemolyticus carrying trh gene was targeted in 34 seafood samples by direct plating and colony hybridization procedure. The trh(+)V. parahaemolyticus could be detected in five of 34 (14.7%) samples and the levels ranged from 5.0 x 10(2) to 3.4 x 10(3) cfu g(-1). Colonies of trh(+)V.parahaemolyticus were isolated from the five positive samples. Forty seafood samples were analysed for trh(+)V. parahaemolyticus by colony hybridization following enrichment in alkaline peptone water. 16 samples (40%) were positive for trh gene and trh(+)V. parahaemolyticus was isolated from 15 samples (37.5%). To assess the sensitivity of the trh probe, seafood homogenates spiked with known concentrations of trh-positive V. parahaemolyticus were plated and hybridized. Counts obtained using the probe were similar to those of inocula. The results suggest that the AP-labelled trh probe is useful for the detection and enumeration of trh(+)V. parahaemolyticus in seafood.