Plates are incubated at 42°C for 48 to 72 hrs in microaerophilic conditions. Inoculated media are incubated at 35°C for 18 — 24 hours. After incubating, 0.1 ml of sample is added to Dynabeads and further incubated at 25°C for 6 hrs on a shaker. They are gel purified using a QIAquick gel extraction kit and sequenced on both strands by DNA Landmarks. Once at the laboratory, a sterile swab is used to obtain a homogenized sample, which is streaked for isolated colonies onto Campylobacter blood-free agar.
Plates are examined for typical E. coli non-sorbitol fermenting colony morphology. Suspect colonies are non-lactose fermenting (NLF) colonies with or without H2S production. This method is based on the MFLP-90 and MFLP-80 Identification of E. coli 0157 by Dynabead™ anti-E. coli 0157 method of the Compendium of Analytical Methods, Health Protection Branch, Methods of Microbiological Analysis of Food, Government of Canada. Agriculture Culture Detection A modification of MFLP-75 primary isolation method of the Compendium of Analytical Methods, Health Protection Branch, Methods of Microbiological Analysis of Food, Government of Canada is used for the detection of Salmonella in manure.
Popoff, Michel, Y. 2001. Antigenic Formulas of the Salmonella Serovars, 8th ed. WHO Collaborating Center for Reference and Research in Salmonella, Institut Pasteur, Paris, France. Method #2 (implemented in 2009)Fifty grams of pork meat is processed in 450 ml of Luria Bertani-Bile Salt Irgasan (LB-BSI) broth, incubated at 12°C for 24 hrs and inoculated with 5ml of Irgasan solution. Once at the laboratory, a sterile swab is used to obtain a homogenized sample, which is streaked for isolated colonies onto Yersinia selective agar (CIN) and incubated at 30°C for 24 hours. After incubation, a loopful of the MacConkey Broth is streaked for isolated colonies onto Sorbitol MacConkey Agar (SMAC) and CT-SMAC Agar plates. Positive samples are then retrieved, and added to Hunts Enrichment Broth (HEB); processed and serial dilutions are prepared.