Abstract

For foodborne outbreak investigations it can be difficult to isolate the relevant strain from food and/or environmental sources. If the sample is contaminated by more than one strain of the pathogen the relevant strain might be missed. In this study mixed cultures of Salmonella enterica were grown in one set of standard enrichment media to see if culture bias patterns emerged. Nineteen strains representing four serogroups and ten serotypes were compared in four-strain mixtures in Salmonella-only and in cattle fecal culture enrichment backgrounds using Salmonella enrichment media. One or more strain(s) emerged as dominant in each mixture. No serotype was most fit, but strains of serogroups C2 and E were more likely to dominate enrichment culture mixtures than strains of serogroups B or C1. Different versions of Rappaport-Vassiliadis (RV) medium gave different patterns of strain dominance in both Salmonella-only and fecal enrichment culture backgrounds. The fittest strains belonged to serogroups C1, C2, and E, and included strains of S. Infantis, S. Thompson S. Newport, S. 6,8:d:-, and S. Give. Strains of serogroup B, which included serotypes often seen in outbreaks such as S. Typhimurium, S. Saintpaul, and S. Schwarzengrund were less likely to emerge as dominant strains in the mixtures when using standard RV as part of the enrichment. Using a more nutrient-rich version of RV as part of the protocol led to a different pattern of strains emerging, however some were still present in very low numbers in the resulting population. These results indicate that outbreak investigations of food and/or other environmental samples should include multiple enrichment protocols to ensure isolation of target strains of Salmonella.

Highlights

  • Enrichment culture is a competition among microbiota for available nutrients and against growth inhibitors

  • The issue of culture bias or culture fitness between strains of the same species or subgroup has been described for Listeria monocytogenes [1,2] and Salmonella [3,4]

  • Culture bias in RV enrichments Strains were sorted into eight groups of four based on similar growth rates in TSB, final cell density in enrichments using RV medium, and those that grew best in RV individually

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Summary

Introduction

Enrichment culture is a competition among microbiota for available nutrients and against growth inhibitors. While enrichment media are designed to favor a target organism the conditions may not favor every strain or sub-group (e.g. serotype, serogroup) of that species. This is of particular concern when the organism being sought is a pathogen from a complex matrix, such as a foodborne pathogen. These results have stimulated questions about the efficiency of recovery of Salmonella from enrichment media containing more than one strain of Salmonella and whether a particular enrichment protocol might bias the types of strains isolated (serotype, serogroup, genotype, etc). Enrichment bias based on serotype has been reported with some protocols for L. monocytogenes [1,2], so it was natural to ask if serotype or serogroup influenced potential enrichment fitness or bias in Salmonella

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