Equipment and Materials

1. See Chapter 1 of FDA BAM for preparation of food homogenate
2. Pipets, 1, 5, and 10 ml, graduated in 0.1 ml units
3. Glass spreading rods (e.g., hockey stick) 3-4 mm diameter with 45-55 mm spreading area
4. Incubators, 30 ± 2°C and 35 ± 2°C
5. Colony counter
6. Marking pen, black felt type
7. Large and small Bunsen burners
8. Wire loops, No. 24 nichrome or platinum wire, 2 mm and 3 mm id
9. Vortex mixer
10. Microscope, microscope slides, and cover slips
11. Culture tubes, 13 × 100 mm, sterile
12. Test tubes, 16 × 125 mm, or spot plate
13. Bottles, 3 oz, sterile
14. Anaerobic jar, BBL GasPak, with H2 + CO2 generator envelopes and catalyst
15. Water bath, 45 ± 2°C (tempering agar)
16. Water bath, 100 ± 2°C (melting Bacara agar prepared in flask)
17. Culture tube racks
18. Staining rack
19. Petri dishes, sterile, 15 × 100 mm

Media and Reagents

1. Bacara agar plates, chromogenic media prepared plates or flasks purchased from AES Chemunex, Cranbury, NJ.
2. Mannitol-egg yolk-polymyxin (MYP) agar plates (M95)
3. Egg yolk emulsion, 50% (M51)
4. Trypticase soy-polymyxin broth (M158)
5. Polymyxin B solutions for MYP agar (0.1%) and trypticase soy-polymyxin broth (0.15%) (see M95 and M158)
6. Phenol red glucose broth (M122)
7. Tyrosine agar (M170)
8. Lysozyme broth (M90)
9. Voges-Proskauer medium (M177)
10. Nitrate broth (M108)
11. Nutrient agar for B. cereus (M113)
12. Motility medium (B. cereus) (M100)
13. Trypticase soy-sheep blood agar (M159)
14. Nitrite detection reagents (R48)
15. Butterfield's phosphate-buffered dilution water (R11) sterilized in bottles to yield final volumes of 450 ± 5 ml and 90 ± 2 ml
16. Voges-Proskauer test reagents (R89)
17. Creatine crystals
18. Gram stain reagents (R32)
19. Basic fuchsin staining solution (R3)
20. Methanol (absolute)
21. Brain Heart Infusion Broth with 0.1% glucose for enterotoxin testing (Ch. 15).