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Orange juice (25 mL)
– Weigh 25 g test portion into 225 mL pre-warmed (35°C) universal pre-enrichment
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broth, C(j), and swirl thoroughly. Let stand at room temperature for 55-65 minutes. Do not mix or adjust
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pH. Incubate, B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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Orange juice (25 mL)
– Weigh 25 g test portion into 225 mL pre-warmed (41°C) BAX® System MP media,
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C(a), and swirl thoroughly. Incubate, B(m), at 39-42°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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Nonfat dry milk (25 g)
– Pour 25 g sample slowly over the surface of 225 mL pre-warmed (35°C) Brilliant
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Green Water, C(c). Let stand at room temperature for 55-65 minutes. Do not mix or adjust pH. Incubate,
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B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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Stainless steel, ceramic tile, plastic –
Add 225 mL pre-warmed (35°C) LB, C(e), to environmental sponge
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in sample bag and swirl thoroughly. Let stand at room temperature for 55-65 minutes. Adjust pH to
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6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if necessary. Incubate, B(m), at 35°C for 22-26 hours.
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Stainless steel, ceramic tile, plastic –
Add 225 mL pre-warmed (35°C) BPW, C(d), to environmental
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sponge in sample bag and swirl thoroughly. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if
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necessary. Incubate, B(m), at 35°C for 18-24 hours.
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Cocoa (25 g)
- Weigh 25 g test portion into sterile container. Use a stomacher, B(n), to homogenize
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sample for 2 minutes with 225 mL reconstituted nonfat dry milk, C(i). Let stand at room temperature for
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55-65 minutes, then swirl thoroughly to mix. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if
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necessary. Add 0.45 mL 1% aqueous brilliant green dye solution, C(l), and mix well. Incubate, B(m), at
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35°C for 22-26 hours. Transfer 10 µL enrichment to 500 µL BHI broth, C(b) before processing.
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White pepper (25 g)
- Weigh 25 g test portion into sterile container. Use a stomacher, B(n), to
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homogenize sample for 2 minutes with 225 mL pre-warmed (35°C) TSB, C(f). Let stand at room
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temperature for 55-65 minutes. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if necessary.
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Incubate, B(m), at 35°C for 22-26 hours.
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Dry Pet Food (375 g)
– Weigh 375 g test portion into sterile container. Use a stomacher, B(n), to
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homogenize sample for 2 minutes with approximately one-third to one-half of 3375 mL pre-warmed
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(35°C) LB, C(e). Add the remainder of the pre-warmed media. Incubate, B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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Dry Pet Food (375 g)
– Weigh 375 g test portion into sterile container. Use a stomacher, B(n), to
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homogenize sample for 2 minutes with approximately one-third to one-half of 3375 mL pre-warmed
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(35°C) BPW, C(d). Add the remainder of the pre-warmed media. Incubate, B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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E.
Re-Growth
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After enrichment, transfer 10 µL of the enrichment to 500 µL BHI broth, C(b). Incubate, B(m), at 37°C for
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3 hours.
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Dupont BAX Salmonella Pre- Collaborative Study
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