AUTHOR=Mangar Preeti , Barman Partha , Kumar Anoop , Saha Aniruddha , Saha Dipanwita 

TITLE=Detection of Virulence-Associated Genes and in vitro Gene Transfer From Aeromonas sp. Isolated From Aquatic Environments of Sub-himalayan West Bengal

JOURNAL=Frontiers in Veterinary Science

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2022.887174

DOI=10.3389/fvets.2022.887174

ISSN=2297-1769

ABSTRACT=<p><italic>Aeromonas</italic> is omnipresent in aquatic environments and cause disease within a wide host range. A total of thirty-four isolates from water samples of small fish farms were identified as <italic>Aeromonas</italic> based on biochemical characteristics and 16S rRNA gene sequence. A total of six virulent factors were analyzed which indicated 100% of isolates as beta-haemolytic and proteolytic, whereas 44.1, 38.2, and 70.6% of isolates produced DNAse, siderophore, and amylase, respectively. Studies on the occurrence of four genetic determinants of virulence factors revealed that <italic>aer/haem</italic> (haemolytic toxin) and <italic>flaA</italic> (polar flagella) genes were present in 44.1% of strains whereas <italic>ascV</italic> (type 3 secretion system) and <italic>aspA</italic> (serine protease) genes were detected in 21.5 and 8.82% of strains, respectively. Fish (<italic>Anabas testudineus</italic>) challenge studies showed that the isolate GP3 (<italic>Aeromonas veronii</italic>) bearing five virulent factors with the combination of <italic>aer/haem</italic><sup>+</sup><italic>/ascV</italic><sup>+</sup><italic>/fla</italic><sup>+</sup> genes induced severe lesions leading to 100% of mortality. In contrast, RB7 possessing four virulence factors and three genes (<italic>aer/haem</italic><sup>+</sup><italic>/ascV</italic><sup>+</sup><italic>/aspA</italic><sup>+</sup>) could not produce severe lesions and any mortality indicating the absence of correlation between the virulence factors, its genes, and the pathogenicity in fishes. GP3 was cytotoxic to human liver cell line (WRL-68) in trypan blue dye exclusion assay. The 431 bp <italic>aer/haem</italic> gene of GP3 was transferable to <italic>E. coli</italic> Dh5α with a conjugational efficiency of 0.394 × 10<sup><bold>–</bold>4</sup> transconjugants per recipient cell. The transfer was confirmed by PCR and by the presence of 23-kb plasmids in both donor and transconjugants. Therefore, the occurrence of mobile genetic elements bearing virulence-associated genes in <italic>Aeromonas</italic> indicates the need for periodic monitoring of the aquatic habitat to prevent disease outbreaks.</p>