AOAC ERP MICRO AUGUST 2018

OMAMAN-31 E: AOAC RI PTM Report 091501 ERP Use Only - March 2016

streaked to XLT4 and BGS agar. Both selective agars were incubated at 35 ± 2 o C for 18-24 hours. In no growth was present, plates were incubated for an additional 24 hours. Presumptive positive Salmonella colonies from each selective agar were picked and transferred to TSI and LIA slants and incubated at 35 ± 2°C for 24 ± 2 hours. Growth from samples producing typical biochemical reactions in TSI and LIA, were streaked to TSA slants and incubated at 35 ± 2 o C for 18-24 hours. Following incubation, isolates were serologically tested for both somatic O and flagellar H agglutination. Additionally, purified TSA isolates were identified using the VITEK ® 2 GN Biochemical Identification card following AOAC Official Method 2011.17. [8]. For the FDA/BAM reference method, all test portions were enriched following the procedures outlined in Chapter 5 of the BAM. Each matrix was evaluated using 25 g test portion sizes. All test portions were homogenized for 2 ±0.2 minutes by stomaching. Following homogenization, test portions were allowed to stand at room temperature (24 ± 2 o C) for 60 ± 5 minutes. If necessary, the pH of the enrichments for all matrixes was adjusted to 6.8 ± 0.2. Subsequently, all Environmental sponges and swabs were enriched with 225 mL and 10 mL of Lactose broth (LB), respectively. Environmental test portions were homogenized by either vortex mixing or by hand massaging and were allowed to stand at room temperature (24 ± 2 o C) for 60 ± 5 minutes. If necessary the pH of the enrichment was adjusted to 6.8 ± 0.2. Subsequently, all enrichments Following incubation, 0.1 mL of primary enrichment was transferred into 10 mL of RV and 1.0 mL into 10 mL of TT broth. RV tubes were incubated at 42 ± 0.2 o C for 24 ± 2 hours. For foods containing low microbial backgrounds (<10 4 ) TT tubes were incubated at 35 ± 2 o C for 24 ± 2 hours. For foods with high microbial backgrounds (>10 4 ) TT tubes were incubated at 43 ± 0.2 o C for 24 ± 2 hours. Following incubation, a loop full of the secondary enrichments were streaked to BS, HE and XLD agar and incubated at 35 ± 2 o C for 24 ± 2 hours. If no visible colonies were present after 24 hours of incubation on the BS plates, they were re-incubated for an additional 24 ± 2 hours at 35 ± 2 o C. A minimum of two suspect colonies from each selective agar were transferred to Triple Sugar Iron agar (TSI) and Lysine Iron agar (LIA) slants and incubated at 35 ± 2°C for 24 ± 2 hours. Following incubation, TSI and LIA slants were examined for typical reactions. Slants producing typical reactions were streaked to TSA and incubated for 35 ± 2°C for 18-24 hours. Following incubation, isolates were serologically tested for both somatic O and flagella H agglutination. Additionally, purified TSA isolates were identified using the VITEK ® 2 GN Biochemical Identification card following AOAC Official Method 2011.17. matrix enrichments were incubated at 35 ± 2°C for 24 ± 2 hours. were incubated at 35 ± 2°C for 24 ± 2 hours. FDA/BAM Chapter 5 Salmonella

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AOAC Research Institute Expert Review Panel Use Only

Guidance for Industry: Sampling and Microbial Testing Of Spent Irrigation Water During

Sprout Production

Test portions of 375 mL were enriched with 3.375 L of BPW containing 40 mg/L novobiocin and were homogenized for 2 ±0.2 minutes. Following homogenization, test portions were allowed to stand at room temperature (24 ± 2 o C) for 60 ± 5 minutes. If necessary, the pH of the