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Luminescent vibriosis is a major bacterial disease in shrimp hatcheries and causes up to 100% mortality in larval stages of penaeid shrimps. We investigated the virulence factors and genetic identity of 29 luminescent  Vibrio  isolates from Indian shrimp hatcheries and farms, which were earlier presumed as  Vibrio harveyi . Haemolysin gene-based species-specific multiplex PCR and phylogenetic analysis of  rpoD  and  toxR  identified all the isolates as  V. campbellii . The gene-specific PCR revealed the presence of virulence markers involved in quorum sensing ( luxM ,  luxS, cqsA ), motility ( flaA ,  lafA ), toxin ( hly, chiA , serine protease, metalloprotease), and virulence regulators ( toxR, luxR ) in all the isolates. The deduced amino acid sequence analysis of virulence regulator ToxR suggested four variants, namely A123Q150 (AQ; 18.9%), P123Q150 (PQ; 54.1%), A123P150 (AP; 21.6%), and P123P150 (PP; 5.4% isolates) based on amino acid at 123rd (proline or alanine) and 150th (glutamine or proline) positions. A significantly higher level of the quorum-sensing signal, autoinducer-2 (AI-2, p = 2.2e−12), and significantly reduced protease activity (p = 1.6e−07) were recorded in AP variant, whereas an inverse trend was noticed in the Q150 variants AQ and PQ. The pathogenicity study in  Penaeus  ( Litopenaeus )  vannamei  juveniles revealed that all the isolates of AQ were highly pathogenic with Cox proportional hazard ratio 15.1 to 32.4 compared to P150 variants; PP (5.4 to 6.3) or AP (7.3 to 14). The correlation matrix suggested that protease, a metalloprotease, was positively correlated with pathogenicity (p > 0.05) and negatively correlated (p < 0.05) with AI-2 and AI-1. The syntenic organization of  toxS-toxR-htpG  operon in  V. campbellii  was found to be similar to pathogenic  V. cholerae  suggesting a similar regulatory role. The present study emphasizes that  V. campbellii  is a predominant pathogen in Indian shrimp hatcheries, and ToxR plays a significant role as a virulence regulator in the quorum sensing—protease pathway. Further, the study suggests that the presence of glutamine at 150th position (Q150) in ToxR is crucial for the pathogenicity of  V. campbellii .

Delineating virulence of Vibrio campbellii: a predominant luminescent bacterial pathogen in Indian shrimp hatcheries