Conventional and molecular methods to detect bacterial pathogens in mussels

Abstract

Aim: To detect Aeromonas spp., Salmonella spp., Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus in mussels and water samples from a farming area, conventional and molecular methods were applied to enrichment cultures. Methods and Results: The aerolysin gene (aero) of Aeromonas spp., the invasion plasmid antigen B (ipaB) gene of Salmonella spp., the enterotoxin secretion protein (epsM) gene of V. cholerae, the species‐specific region of 16S rRNA gene of V. vulnificus, the 16S–23S rDNA (IGS) gene of V. parahaemolyticus and the pR72H fragment of V. parahaemolyticus were amplified by multiplex polymerase chain reaction (PCR) assays on DNA extracted from enrichment cultures. The haemolysin gene (tdh) of pathogenic V. parahaemolyticus was also amplified. Conventional culture method allowed the isolation of V. parahaemolyticus and V. vulnificus from water and mussels. The genes aero, epsM and 16S rRNA of V. vulnificus were occasionally detected in the enrichment cultures. In mussels, the ipaB and IGS genes were detected from June to September and from April to November, respectively. All genes, except aero, were amplified from mussels collected in September, when pathogenic V. parahaemolyticus (tdh+) strains were also isolated. Conclusions: Multiplex‐PCR assays were more sensitive and faster than conventional procedures. Significance and Impact of the Study: The results emphasize the need of an accurate and rapid detection of bacterial pathogens in mussels to protect human health.

A study on the prevalence of Aeromonas spp. and its enterotoxin genes in samples of well water, tap water, and bottled water

“…In this study, in agreement with Gugliandolo et al . [ 31 ] and Liu [ 32 ], it is revealed that PCR ( Figure-4 ) was a better, sensitive, and cheap method compared to cultural assay (90% compared to PCR). Balakrishna et al .…”

Preliminary Study on the In vitro and In vivo Effects of Asparagopsis taxiformis Bioactive Phycoderivates on Teleosts

“…piscicida (Pdp), Vibrio alginolyticus (Ag37), V. harveyi (G5), V. parahaemolyticus (L12G), and V. vulnificus (OPV10) which are part of the Culture Collection housed at the Dept of Chemistry, Biology, Pharmaceutical and Environmental Sciences of the University of Messina (Italy). The strains were previously isolated and identified as reported in Gugliandolo et al (2008, 2011). They were maintained at −20°C in Tryptone Soy Broth (Difco) supplemented with 1% (w/v) NaCl (TSB1) and 50% (v/v) glycerol.…”

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“…M-PCR has also been widely used for the detection of Vibrio spp. Gugliandolo et al (2010) compared conventional and molecular methods in detection of Aeromonas spp., Salmonella spp., Vibrio cholera, Vibrio parahaemolyticus and Vibrio vulnificus in mussels and water samples from a farming area. At the end of the research they found that conventional methods were less sensitive than multiplex PCR method; conventional enrichment and isolation procedures allowed only the isolation of V. parahaemolyticus and V. vulnificus strains, while Aeromonas spp., Salmonella spp.…”

“…It is particularly important when a risk assessment is required for public health if a first sign of an illness occur less than 24 hours after ingestion of contaminated seafood (Gugliandolo, Lentini, Spano, & Maugeri 2010).…”