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Comparison of methods to detect low levels of Salmonella enterica in surface waters to support antimicrobial resistance surveillance efforts performed in multiple laboratories
,Identifying and developing effective and sensitive detection methods for antimicrobial resistant Salmonella enterica from surface water is a goal of the U.S. National Antimicrobial Resistance Monitoring System (NARMS). No specific microbiological methods used in surveillance efforts for Salmonella enterica or antimicrobial resistant S. enterica in water have been standardized or reported in the U.S. Here we describe a multi-laboratory evaluation of four methods, bulk water enrichment (BW), vertical Modified Moore Swab (VMMS), modified Standard Method 9260.B3 (SM), and dead-end ultrafiltration (DEUF), to recover S. enterica from surface water. In Phase 1, one-liter volumes of surface water (n=60) were collected from the same site in Fall 2021 on five different dates. Water was shipped and analyzed at four different USDA ARS laboratories for recovery of environmental Salmonella and an inoculated fluorescent S. Typhimurium strain (ca. 30 CFU/L). One-liter samples (n=20) were subjected to recovery and enrichment by either BW, VMMS, or SM. Overall, fluorescent S. Typhimurium and environmental Salmonella spp. were recovered from 65% (39/60) and 45% (27/60) of water samples, respectively. SM, VMMS, and BW recovered fluorescent S. Typhimurium from 75%, 60% and 60% of inoculated samples, respectively. Analysis by Chi-squared test determined that laboratory location had a significant (p < 0.05) effect on recovery compared to method or date of water collection. In Phase 2, DEUF was compared to SM at two different laboratory locations to recover fluorescent S. Typhimurium (30 CFU/L) from 1-L samples. SM and DEUF recovered S. Typhimurium from 100% (20/20) and 95% (19/20) of inoculated water samples, respectively; laboratory location nor recovery method (p> 0.05) affected S. Typhimurium recovery. Results indicate that SM method consistently recovered low levels of Salmonella from inoculated water samples and should be prioritized for Salmonella recovery from surface water.,Resources in this dataset:,,
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Data from: Detection of Salmonella enterica and Listeria monocytogenes in alternative irrigation water by culture and qPCR-based methods in the Mid-Atlantic U.S.
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,Alternative irrigation waters (rivers, ponds, and reclaimed water) can harbor bacterial foodborne pathogens like Salmonella enterica and Listeria monocytogenes, potentially contaminating fruit and vegetable commodities. Detecting foodborne pathogens using qPCR-based methods may accelerate testing methods and procedures compared to culture-based methods. This study compared detection of S. enterica and L. monocytogenes by qPCR (real-time PCR) and culture methods in irrigation waters to determine the influence of water type (river, pond, and reclaimed water), season (winter, spring, summer, and fall), or volume (0.1, 1, and 10 L) on sensitivity, accuracy, specificity, and positive (PPV), and negative (NPV) predictive values of these methods. Water samples were collected by filtration through modified Moore swabs (MMS) over a 2-year period at 11 sites in the Mid-Atlantic U.S. on a bi-weekly or monthly schedule. For qPCR, bacterial DNA from culture-enriched samples (n = 1,990) was analyzed by multiplex qPCR specific for S. enterica and L. monocytogenes. For culture detection, enriched samples were selectively enriched, isolated, and PCR confirmed. PPVs for qPCR detection of S. enterica and L. monocytogenes were 68% and 67%, respectively. The NPV were 87% (S. enterica) and 85% (L. monocytogenes). Higher levels of qPCR/culture agreement were observed in spring and summer compared to fall and winter for S. enterica; for L. monocytogenes, lower levels of agreement were observed in winter compared to spring, summer, and fall. Reclaimed and pond water supported higher levels of qPCR/culture agreement compared to river water for both S. enterica and L. monocytogenes, indicating that water type may influence the agreement of these results.,
Sampling and Analysis of Antimicrobial Resistant Bacteria in Surface Water Samples Collected in the East Fork Watershed in Southwest Ohio Dataset
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Dataset for a year long watershed study in southwestern Ohio to analyze antimicrobial resistant bacteria (E. coli, Enterococcus, and Salmonella) in surface waters and wastewater treatment plant effluent.
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