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A multipathogen selective enrichment broth for simultaneous growth of Salmonella enterica serovar Enteritidis, Staphylococcus aureus, and Listeria monocytogenes

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Abstract:

A selective enrichment broth (SSL) was formulated to allow concurrent growth of 3 prominent food-borne pathogens: Salmonella enterica serovar Enteritidis, Staphylococcus aureus, and Listeria monocytogenes. Nalidixic acid, lithium chloride, and potassium tellurite were added as the selective agents, while sodium pyruvate and mannitol were employed as the supplemented elements. In the individual growth trial, the target pathogens were capable of growing in SSL to as high as 7-8 log10 colony-forming units (CFU)/mL after 24 h incubation at 37 °C when being inoculated at 50-100 CFU/mL. In the simultaneous growth trial, the 3 combined target pathogens showed similar growth rates. The results show that SSL could support the successful simultaneous enrichment of 3 pathogens; however, SSL inhibited the growth of nontarget bacteria. In the artificial contaminated raw beef and ready-to-eat chicken, a high recovery of these 3 target pathogens was obtained in SSL. Finally, Salmonella Enteritidis, Staphylococcus aureus, and L. monocytogenes were detected from 710 suspicious food samples by SSL with real-time PCR, and no false-positive or -negative results were reported. In summary, SSL has been shown to be a suitable broth for the simultaneous detection of the 3 prominent food-borne pathogens by multipathogen detection on a single-assay platform.

Un milieu de culture sélectif enrichi (SSL, selective enrichment broth) a été formulé afin de permettre une croissance concomitante de 3 pathogènes alimentaires importants: Salmonella enterica sérotype Enteritidis, Staphylococcus aureus et Listeria monocytogenes. L’acide nalidixique, le chlorure de lithium et le tellurate de potassium ont été ajoutés comme agents sélectifs, alors que le pyruvate de sodium et le mannitol ont été utilisés comme suppléments. Lors de tests de croissance individuels, les pathogènes cibles étaient capables de croître dans le SSL à des niveaux allant jusqu’à 7-8 log10 UFC/mL après 24 h d’incubation à 37 °C lorsqu’ils étaient inoculés à une concentration de 50-100 UFC/mL. Lors de tests de croissance simultanée, les 3 pathogènes combinés montraient des taux de croissance similaires. Les résultats ont montré que le SSL permettait un bon enrichissement des 3 pathogènes tout en inhibant la croissance de bactéries non ciblées. Une bonne récupération de ces 3 pathogènes à partir de bœuf cru et de poulet prêt-à-servir contaminés artificiellement a été obtenue dans le milieu SSL. Finalement, Salmonella Enteritidis, Staphylococcus aureus et Listeria monocytogenes ont été détectés dans 710 échantillons alimentaires suspects dans le milieu SSL par une PCR en temps réel, et aucun faux positif ou négatif n’a été rapporté. En résumé, le SSL s’est avéré un bon milieu de culture pour détecter simultanément 3 pathogènes alimentaires importants sur une plateforme de détection de pathogènes multiples à l’aide d’un test unique.

Document Type: Research Article

Publication date: 2010-07-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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