Abstract
The objective of this study was to investigate the virulence factors, genetic relationship, antibiotic resistance profile and the biofilm formation ability of Vibrio parahaemolyticus isolates on shrimp and mussel surfaces at 30°C. In this study, eight (n = 8) V. parahaemolyticus isolated from mussel were examined. We used the polymerase chain reaction (PCR) to examine the distribution of different genes, and Repetitive Extragenic Palindromic-PCR (REP-PCR) to compare the genetic relationship. Disk diffusion technique was used to assess antibiotic and multiple-antibiotic resistance. The biofilm formation assay, and field emission scanning electron microscopy (FE-SEM) were used to evaluate biofilm formation ability. Transmission Electron Microscope (TEM) was used to observe the morphological structure of bacterial cell. Our results indicated that the biofilm-associated genes, 16S rRNA, toxR, and tdh, were present in all the tested V. parahaemolyticus isolates (n = 8). Approximately, 62.5% (5 isolates among 8 isolates) isolates showed strong multiple-antibiotic resistance index with an average value of 0.56. All isolates (n = 8) showed strong genetic relationship and significant biofilm formation ability on shrimp and mussel surfaces. This study demonstrated that the presence of virulence factors, high multiple antibiotic resistance index (MARI) values, and effective biofilm formation ability of V. parahaemolyticus isolates could be a great threat to human health and economic values in future. It was also suggested that a high resistance rate to antibiotic could be ineffective for treating V. parahaemolyticus infections. The continuous monitoring of V. parahaemolyticus antibiotic, molecular and biofilm characteristics is needed to increase seafood safety.
Highlights
Seafood is recognized as a nutritious and healthy food choice, and is accepted by increasing numbers of consumers worldwide (Hellberg et al, 2012)
Our results indicated that 87.5% (n = 8) of V. parahaemolyticus isolates harbored the complete type three secretion T3SS (VP1690), tox-RS/Old, and VPaI-6 (VP1263); 50% (n = 8) of isolates harbored the complete Type I pilus (VP1506) and type VI secretion T6SS (VP1418) genes
Following detection on the CHROMagar Vibrio plates, polymerase chain reaction (PCR) assays for toxR, and genes associated with biofilm formation and pathogenicity were conducted for molecular characterization of the V. parahaemolyticus isolates
Summary
Seafood is recognized as a nutritious and healthy food choice, and is accepted by increasing numbers of consumers worldwide (Hellberg et al, 2012). The main obstacles in the consumption of seafood are their high perishability and health risk due to contamination by pathogens (Reyhanath and Kutty, 2014). Vibrio parahaemolyticus is the most prevalent shrimp pathogen encountered in aquaculture, causes in shrimp Vibriosis with the potential for severe health crisis (Mohammad et al, 2005; Kleter et al, 2009; Sani et al, 2013; Zhang et al, 2014). During 1997 to 2010, Global production of mussels has increased up to 1.9 million tons worldwide. This represented 95% of the world mussel production, in comparison to 83% in 1997 (Ferreira et al, 2014). As mussel is a good vehicle for Vibrio species, V. parahaemolyticus can survive in mussel with potential contamination (Mannas et al, 2014). Several post-harvest processes, including lowtemperature pasteurization and irradiation have been developed for reducing Vibrios in aquaculture but they are expensive (Chae et al, 2009)
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