| Summary: | A fraction of human <i>Salmonella</i> infections is associated with direct contact with reptiles, yet the number of reptile-associated Salmonellosis cases are believed to be underestimated. Existing data on <i>Salmonella</i> spp. transmission by reptiles in Portugal is extremely scarce. The aim of the present work was to evaluate the prevalence of <i>Salmonella</i> spp. in pet reptiles (snakes, turtles, and lizards), as well as evaluate the isolates’ antimicrobial resistance and virulence profiles, including their ability to form biofilm in the air-liquid interface. Additionally, the antimicrobial effect of chlorhexidine gluconate on the isolates was tested. <i>Salmonella</i> was isolated in 41% of the animals sampled and isolates revealed low levels of antimicrobial resistance. Hemolytic and lypolytic phenotypes were detected in all isolates. The majority (90.63%) of the <i>Salmonella</i> isolates were positive for the formation of pellicle in the air-liquid interface. Results indicate chlorhexidine gluconate is an effective antimicrobial agent, against the isolates in both their planktonic and biofilm forms, demonstrating a bactericidal effect in 84.37% of the <i>Salmonella</i> isolates. This study highlights the possible role of pet reptiles in the transmission of non-typhoidal <i>Salmonella</i> to humans, a serious and increasingly relevant route of exposure in the <i>Salmonella</i> public health framework.
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