Conventional and molecular methods to detect bacterial pathogens in mussels.
OBJECTIVE
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.


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.


CONCLUSIONS
The results emphasize the need of an accurate and rapid detection of bacterial pathogens in mussels to protect human health.
Dipartimento di Biologia Animale ed Ecologia Marina, Università di Messina, Messina, Italy. cgugliandolo@unime.it
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Gugliandolo
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Lentini
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Spanò
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Maugeri
D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D018407:Bacterial Physiological Phenomena; D015373:Bacterial Typing Techniques; D049872:Bivalvia / Q000382:microbiology*; D005798:Genes, Bacterial / Q000235:genetics; D016133:Polymerase Chain Reaction; D017747:Seafood / Q000382:microbiology; D012623:Seawater / Q000382:microbiology; D012680:Sensitivity and Specificity