Stool Cultures for Water Samples Julie A. Ribes, M.D., Ph.D. UKHealthCare Clinical Microbiology Laboratory

What is Routine for Stool Cultures? Salmonella spp. Shigella spp. Direct plating and by Selinite Broth enrichment Edwardsiella tarda 2

What is Routine for Stool Cultures? Campylobacter detection Campy plate with Microaerophilic incubation 3

What is Routine for Stool Cultures? Aeromonas sp. Plesiomonas sp. Bacillus cereus All of these can be identified off of a BAP, MAC or an XLD Aeromonas B. cereus 4

What is Routine for Stool Cultures? Toxigenic E. coli CDC now recommends that testing for E. coli 0157H7 and the other toxigenic E. coli species be performed on all patients being evaluated for diarrheal disease, not just patients with bloody diarrhea Sorbitol MAC and Toxin testing both should be performed 5

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Testing for Giardia and Cryptosporodium Direct Fluorescence Antibody staining (Meridian Biosciences)

Wolf Run Water Findings D13 BAP Sweep oxidase to pick up Aeromonas Pleisiomonas Spready colony worked up for possible Bacillus cereus

Wolf Run Water Findings D13 MAC Screen all clear colony types (non-lactose fermenters {NLF}) for possible Salmonella and Shigella Usual stools, not too many NLF

Wolf Run Water Findings D13 XLD Usual coliforms are yellow Need to screen black colonies for possible Salmonella spp. Need to screen clear red colonies for possible Shigella spp.

Wolf Run Water Findings D13 MAC Sorbitol Pink colonies are sorbitol fermenters (usual flora) Screen all clear colonies for Shiga toxin-producing E coli 0157H7

Shiga toxin testing on Overnight Broth

Wolf Run Water Findings D13 CIN Agar Usually not much growth Need to screen “bulls eye” colonies with pink centers

Potentially Significant Organisms Isolated D13 8/24/10 Bacillus cereus, Aeromonas sobria, Pleisiomonas shigelloides D10 8/24/10 Aeromonas sobria, Bacillus cereus D14 8/24/10 Bacillus cereus, Aeromonas sobria D16 8/24/10 Aeromonas caviae, Aeromonas sobria, Bacillus cereus D18 8/24/10 Bacillus cereus, Aeromonas hydrophila, Aeromonas schubertii, Aeromonas sobria, Aeromonas caviae

Potentially Significant Organisms Isolated D03 8/31/10 Aeromonas hydrophila D13 8/31/10 Aeromonas sp., Aeromonas sobria. Bacillus cereus D18 8/31/10 Aeromonas sobria, Pleisiomonas shigelloides D19 8/31/10 Aeromonas sp., Aeromonas hydrophila D25 8/31/10 Aeromonas sobria, Aeromonas caviae

Wolf Run Water Findings D25 BAP No spready colonies Still had some Oxidase positive organisms to work up

Wolf Run Water Findings D25 MAC Pink and clear colonies Still needed to look at the clear ones

Wolf Run Water Findings D25 XLD All yellow colonies, nothing to work up for Enterobacteraceae

Wolf Run Water Findings D25 CNA Smattering of Gram positives, nothing significant

Aeromonas hydrophila Aeromonas caviae

Citrobacter freundii

Bacillus cereus