Identification of Vibrio parahaemolyticus from farmed penaeid shrimp, Penaeus vannamei (Boone, 1931) by multiplex PCR targeting toxR and tlh genes

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Targeted surveillance was conducted in 37 Penaeus vannamei farms in Tamil Nadu, Indiato find out the prevalence of Vibrio parahaemolyticus (Vp) infections. Special emphasis wasgiven to screen the isolates for acute hepatopancreatic necrosis disease (AHPND). Bacteriawere isolated from haemolymph, stomach and hepatopancreas. Initial identificationswere carried out using the morphological, physiological and biochemical characteristics.Vp isolates were further confirmed by a newly developed multiplex PCR targeting Vp specifictoxR and tlh genes. PCR screening was carried out for AHPND causing AP1, AP2, AP3(pirAvp, Photorhabdus insect-related binary toxin gene) and AP4 (pirAvp and pirBvp) genes.In addition, PCR was also performed for human pathogenic tdh and trh genes. The PCRresults revealed that there were 26 (35.14%) isolates affirmative for Vp specific toxR and tlhgenes and negative for AP1, AP2, AP3 and AP4 genes indicating that there was no AHPNDcausing Vp strain among the isolates. The isolates were also negative for anthropozoonotictdh and trh genes. The multiplex PCR developed targeting Vp specific toxR and tlh geneswould be a useful technique for easy identification of Vp strains from shrimp farms. Keywords:Acute hepatopancreatic necrosis disease (AHPND), Early mortality syndrome (EMS), Multiplex PCR, Vibriosis, Vibrio parahaemolyticus (Vp)

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