A. Equipment and Materials

1. Blender and sterile blender jars
2. Sterile, homogenizer blender filter bags (710 ml) or equivalent, sterile 500 ml wide mouth, screw-cap jars, sterile 500 ml Erlenmeyer flasks, sterile 250 ml beakers, sterile glass or paper funnels of appropriate size, and, optionally, containers of appropriate capacity to accommodate large and composited samples
3. Sterile, bent glass or plastic spreader rods
4. Balance, 2000 g capacity, sensitivity of 0.1 g
5. Balance, 120 g capacity, sensitivity of 5 mg
6. Incubator, 35 ± 2°C
7. Refrigerated incubator or laboratory refrigerator, 4 ± 2°C
8. Water bath, 49 ± 1°C
9. Water bath, circulating, thermostatically-controlled, 42 ± 0.2°C and 43 ± 0.2°C
10. Sterile spoons or other appropriate instruments for transferring food samples
11. Sterile culture dishes, 15 × 100 mm, glass or plastic
12. Sterile pipets, 1 ml, with 0.01 ml graduations; 5 and 10 ml, with 0.1 ml graduations
13. Sterile inoculating needle and inoculating loop (10 µl and 1 µl)
14. Sterile test or culture tubes, 16 × 150 mm and 20 × 150 mm; serological tubes, 10 × 75 mm or 13 × 100 mm
15. Test or culture tube racks
16. Vortex mixer
17. Sterile shears, large scissors, scalpel, and forceps
18. Fisher or Bunsen burner
19. pH test paper (pH range 6-8) with maximum graduations of 0.4 pH units per color change 22. pH meter
20. Plastic bags, 28 × 37 cm, sterile, with resealable tape (Items 21-22 are needed in the analysis of frog legs and rabbit carcasses.)
21. Plastic beakers, 4 liter, autoclavable, for holding plastic bag during shaking and incubation
22. Sponges, non-bactericidal, or equivalent
23. Swabs, non-bactericidal, cotton-tipped
24. Ultraviolet (UV) light source with an emission wavelength between 360 to 400 nm when Green Fluorescent Protein (GFP)-UV control strains are used.

B. Media, Reagents, and Quality Control Cultures

1. Lactose broth (M74)
2. Nonfat dry milk (reconstituted) (M111)
3. Selenite cystine (SC) broth (M134)
4. Tetrathionate (TT) broth (M145)
5. Rappaport-Vassiliadis (RV) medium (M132). NOTE: RV medium must be made rom its individual ingredients. Commercial formulations are not acceptable.
6. Xylose lysine desoxycholate (XLD) agar (M179)
7. Hektoen enteric (HE) agar (M61)
8. Bismuth sulfite (BS) agar (M19)
9. Triple sugar iron agar (TSI) (M149)
10. Tryptone (tryptophane) broth (M164)
11. Trypticase (tryptic) soy broth (M154)
12. Trypticase soy-tryptose broth (M160)
13. MR-VP broth (M104)
14. Simmons citrate agar (M138)
15. Urea broth (M171)
16. Urea broth (rapid) (M172)
17. Malonate broth (M92)
18. Lysine iron agar (LIA) (Edwards and Fife) (M89)
19. Lysine decarboxylase broth (M87)
20. Motility test medium (semisolid) (M103)
21. Potassium cyanide (KCN) broth (M126)
22. Phenol red carbohydrate broth (M121)
23. Purple carbohydrate broth (M130)
24. MacConkey agar (M91)
25. Nutrient broth (M114)
26. Brain heart infusion (BHI) broth (M24)
27. Papain solution, 5% (M56a)
28. Cellulase solution, 1% (M187)
29. Tryptose blood agar base (M166)
30. Universal preenrichment broth (M188)
31. Universal preenrichment broth (without ferric ammonium citrate) (M188a)
32. Buffered peptone water (M192)
33. Dey-Engley broth (M193)
34. Potassium sulfite powder, anhydrous
35. Chlorine solution, 200 ppm, containing 0.1% sodium dodecyl sulfate (R12a)
36. Ethanol, 70% (R23)
37. Kovacs' reagent (R38)
38. Voges-Proskauer (VP) test reagents (R89)
39. Creatine phosphate crystals
40. Potassium hydroxide solution, 40% (R65)
41. 1 N Sodium hydroxide solution (R73)
42. 1 N Hydrochloric acid (R36)
43. Brilliant green dye solution, 1% (R8)
44. Bromcresol purple dye solution, 0.2% (R9)
45. Methyl red indicator (R44)
46. Sterile distilled water
47. Tergitol anionic 7 (R78)
48. Triton X-100 (R86)
49. Physiological saline solution, 0.85% (sterile) (R63)
50. Formalinized physiological saline solution (R27)
51. Salmonella polyvalent somatic (O) antiserum
52. Salmonella polyvalent flagellar (H) antiserum
53. Salmonella somatic group (O) antisera: A, B, C1, C2, C3, D1, D2, E1, E2, E3, E4, F, G, H, I, Vi, and other groups, as appropriate
54. Salmonella Spicer-Edwards flagellar (H) antisera
55. Modified buffered peptone water, two formulas (M192b, M192c)

Suggested Quality Control Cultures

Analysts must use both typical and atypical Salmonella controls for sample analysis, since it may be more difficult to confirm the presence of an atypical strain in a sample without an atypical control strain for guidance.

Green fluorescent protein (GFP) control strains which fluoresce under UVA light, more specifically at wavelengths between 360 to 400 nm, have been developed by FDA and have been licensed by FDA to Microbiologics for distribution. (200 Cooper Avenue North St. Cloud, MN 56303, 1800-599-2847). The following cultures may be purchased from Microbiologics:

• Salmonella enterica subsp. enterica serovar Gaminara Sal57 / FDA SAL5695 (typical, lactose negative, H2S positive), Catalog No. 01278UV.

• Salmonella enterica subsp. enterica serovar Senftenberg Sal59 / FDA Sal5697 (atypical, lactose negative, H2S negative), Catalog No. 01226UV.

• Salmonella enterica subsp. enterica serovar Mbandaka Sal58 / FDA SAL5696 (atypical, sucrose positive, H2S positive), Catalog No. 01230UV.

Non-GFP fluorescent Salmonella control strains may also be used for sample analysis. In addition to the positive control cultures (typical Salmonella; lactose negative, H2S positive), 3 additional Salmonella cultures are recommended to assist in the selection of atypical Salmonella colony morphology on selective agars. These cultures are a lactose positive, H2S-positive S. diarizonae (47:i:z53:z57, ATCC 12325) and a lactose-negative, H2S-negative S. Abortusequi (4,12:-:e,n,x, ATCC 9842); or a lactose-positive, H2S-negative S. diarizonae (60:r:e,n,x,z15, ATCC 29934). These cultures may be obtained from the American Type Culture Collection (http://www.atcc.org/), 10801 University Boulevard, Manassas, VA 20110-2209.