美國FDA細(xì)菌學(xué)分析手冊第八版(BAM)沙門氏菌(XXXX版)

1、.：.；BAM: SalmonellaNOTICE:If you are looking for BAM Chapter 5: Salmonella (December 2007 Edition) that is incorporated by reference in 21 CFR Parts 16 and 118: HYPERLINK /Food/FoodSafety/Product-SpecificInformation/EggSafety/EggSafetyActionPlan/ucm170615.htm t Federal Register Final Rule1 (J

2、uly 9, 2021, 74 FR 33030): Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation, please use theseversions of the BAM HYPERLINK /downloads/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM244774.pdf t Salmonella Chapter2

3、(PDF, 189 Kb)and HYPERLINK /downloads/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM244777.pdf t Appendix 1: Rapid Methods for Detecting Foodborne Pathogens3 (PDF, 195 Kb).The most recentEdition of BAM Chapter 5: Salmonella is available below this notice.Novembe

4、r 2021 VersionBacteriological Analytical ManualChapter 5SalmonellaAuthors: Wallace H. Andrews and HYPERLINK mailto:Thomas.H Thomas HammackRevision History:November 2021 - Addition to Section C: Preparation of foods for isolation of Salmonella: Leafy green vegetables and herbs. Febru

5、ary 2021 Removed link to Appendix 1: Rapid Methods for Detecting Foodborne Pathogens (now archived). December 2007 Mamey pulp method added, and Section D revised. June 2006 Eggs method revised for shell eggs and liquid whole eggs. April 2003 Frog legs method, Lactic casein, Rennet casein, Sodium cas

6、einate and Rabbit carcass methods revised, top ears and other dog chew toys added. Removed section A.25, Mechanical shaker. October 25, 2001 Extension of the applicability of the orange juice method in section C.19 to apple juice and apple cider. 1999-DEC, 2000-MAR, and 2000-AUG Final revision on 20

7、00-NOV-14 (see the Introduction for a summary of changes).To obtain a copy of a prior version not currently posted, please contact HYPERLINK mailto:Thomas.H Thomas HammackChapter Contents HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm

8、070149.htm l Intro#Intro Introduction HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Equip#Equip Equipment and Materials HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Media#Medi

9、a Media and Reagents HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Prep#Prep Preparation of foods for isolation of Salmonella HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Isol

10、#Isol Isolation of Salmonella HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Id#Id Identification of Salmonella HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Refs#Refs Reference

11、sIntroductionSeveral changes are being introduced in this edition of BAM (8th Edition). The first change involves the expanded use of HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063568.htm Rappaport-Vassiliadis (RV) medium4 for foods with both high

12、and low levels of competitive microflora. In the previous edition, RV medium was recommended only for the analysis of shrimp. Based on the completion of AOAC precollaborative ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Refs#Refs 5, 6)

13、 and collaborative ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Refs#Refs 7, 8) studies, RV medium is now being recommended for the analysis of high microbial and low microbial load foods. RV medium replaces selenite cystine (SC) broth

14、 for the analysis of all foods, except guar gum. In addition, RV medium replaces lauryl tryptose broth for use with dry active yeast. HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063686.htm Tetrathionate (TT)5 broth continues to be used as the second

15、 selective enrichment broth. However, TT broth is to be incubated at 43C for the analysis of high microbial load foods and at 35C for the analysis of low microbial load foods, including guar gum.The second change involves the option of refrigerating incubated preenrichments and selective enrichments

16、 of low-moisture foods for up to 72 h. With this option, sample analyses can be initiated as late as Wednesday or Thursday without weekend work being involved.The third change involves reducing the period of incubation of the HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAn

17、alyticalManualBAM/ucm064484.htm lysine iron agar (LIA)6 slants. In the former edition (BAM-7), HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063699.htm triple sugar iron agar (TSI)7 and LIA slants were incubated at 35C for 24 2 h and 48 2 h, respectiv

18、ely. Unpublished data have demonstrated that the 48 h reading of LIA slants is without diagnostic value. Of 193 LIA slants examined, all gave definitive results within 24 2 h of incubation. No significant changes altered the final test result when the slants were incubated an additional 24 h. Thus,

19、both the TSI and LIA slants are now incubated for 24 2 h.The fourth change involves the procedure for surface disinfection of shell eggs. In the previous edition (BAM-7), egg shells were surface-disinfected by soaking in 0.1% mercuric chloride solution for 1 h followed by soaking in 70% ethanol for

20、30 min. Mercuric chloride is classified as a hazardous waste, and is expensive to dispose of according to Environmental Protection Agency guidelines. In this edition (BAM-8) egg shells are now surface-disinfected by soaking for at least 10 sec in a 3:1 solution consisting of 3 parts of 70% alcohol (

21、ethyl or isopropyl) to 1 part of iodine/potassium iodide solution.The fifth change involves the sample preparation of eggs. Egg contents (yolk and albumen) are thoroughly mixed before analysis. After mixing the egg contents, 25 g (ml) are added to 225 ml trypticase (tryptic) soy broth supplemented w

22、ith ferrous sulfate.A method for the analysis of guar gum has been included. When guar gum is preenriched at a 1:9 sample/broth ratio, a highly viscous, nonpipettable mixture results. Addition of the enzyme cellulase to the preenrichment medium, however, results in a readily pipettable mixture.A met

23、hod for orange juice (pasteurized and unpasteurized) has been included due to recent orange juice-related outbreaks.The directions for picking colonies from the selective plating agars have been made more explicit to reflect the intent of the method. In the absence of typical or suspect colonies on

24、the selective plating agars, it is recommended that atypical colonies be picked to TSI and LIA slants. This recommendation is based on the fact that up to 4% of all Salmonella cultures isolated by FDA analysts from certain foods, especially seafoods, during the past several years have been atypical.

25、Finally, since the publication of BAM-7, a 6-way comparison was conducted of the relative effectiveness of the three selective plating agars recommended in the BAM ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063348.htm bismuth sulfite8, HYPERLINK

26、/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064342.htm Hektoen enteric9, and HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062986.htm xylose lysine desoxycholate agars10) and three relatively new agars (EF-18,

27、xylose lysine Tergitol 4, and Rambach agars). Our results (9) indicated no advantage in replacing any of the BAM-recommended agars with one or more of the newer agars. Thus, the combination of selective plating agars recommended in BAM-7 remains unchanged.Equipment and Materials Blender and sterile

28、blender jars (see HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063335.htm Chapter 111) Sterile, 16 oz (500 ml) wide-mouth, screw-cap jars, sterile 500 ml Erlenmeyer flasks, sterile 250 ml beakers, sterile glass or paper funnels of appropriate size, a

29、nd, optionally, containers of appropriate capacity to accommodate composited samples Sterile, bent glass or plastic spreader rods Balance, with weights; 2000 g capacity, sensitivity of 0.1 g Balance, with weights; 120 g capacity, sensitivity of 5 mg Incubator, 35 2 C Refrigerated incubator or labora

30、tory refrigerator, 4 2C Water bath, 49 1C Water bath, circulating, thermostatically-controlled, 43 0.2C Water bath, circulating, thermostatically-controlled,42 0.2C Sterile spoons or other appropriate instruments for transferring food samples Sterile culture dishes, 15 x 100 mm, glass or plastic Ste

31、rile pipets, 1 ml, with 0.01 ml graduations; 5 and 10 ml, with 0.1 ml graduations Inoculating needle and inoculating loop (about 3 mm id or 10 5l), nichrome, platinum-iridium, chromel wire, or sterile plastic Sterile test or culture tubes, 16 x 150 mm and 20 x 150 mm; serological tubes, 10 x 75 mm o

32、r 13 x 100 mm Test or culture tube racks Vortex mixer Sterile shears, large scissors, scalpel, and forceps Lamp (for observing serological reactions) Fisher or Bunsen burner pH test paper (pH range 6-8) with maximum graduations of 0.4 pH units per color change pH meter Plastic bags, 28 x 37 cm, ster

33、ile, with resealable tape. (Items 23-24 are needed in the analysis of frog legs and rabbit carcasses.) Plastic beakers, 4 liter, autoclavable, for holding plastic bag during shaking and incubation. Sponges, non-bactericidal (Nasco cat # B01299WA), or equivalent. Swabs, non-bactericidal, cotton-tippe

34、d. HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm055778.htm Media12 and HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm055791.htm Reagents13For preparation of media and reagents, refer to Methods 967.25-

35、967.28 in Official Methods of Analysis ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm l Refs#Refs 1).Lactose broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064414.htm M7414) Nonfat dr

36、y milk (reconstituted) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064144.htm M11115) Selenite cystine (SC) broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063597.htm M13416) Tetrathionate (TT

37、) broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063686.htm M14517) Rappaport-Vassiliadis (RV) medium ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063568.htm M13218). NOTE: RV medium must be m

38、ade from its individual ingredients. Commercial formulations are not acceptable. Xylose lysine desoxycholate (XLD) agar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062986.htm M17919) Hektoen enteric (HE) agar ( HYPERLINK /Food/ScienceResear

39、ch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064342.htm M6120) Bismuth sulfite (BS) agar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063348.htm M1921) Triple sugar iron agar (TSI) ( HYPERLINK /Food/ScienceResearch/LaboratoryMe

40、thods/BacteriologicalAnalyticalManualBAM/ucm063699.htm M14922) Tryptone (tryptophane) broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063867.htm M16423) Trypticase (tryptic) soy broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Ba

41、cteriologicalAnalyticalManualBAM/ucm063805.htm M15424) Trypticase soy broth with ferrous sulfate ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063816.htm M18625) Trypticase soy-tryptose broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethod

42、s/BacteriologicalAnalyticalManualBAM/ucm063862.htm M16026) MR-VP broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064068.htm M10427) Simmons citrate agar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBA

43、M/ucm063639.htm M28) Urea broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062935.htm M17129) Urea broth (rapid) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062967.htm M17230) Malonate broth (

44、HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064500.htm M9231) Lysine iron agar (LIA) (Edwards and Fife) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064484.htm M8932) Lysine decarboxylase broth ( H

45、YPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064474.htm M87)33 Motility test medium (semisolid) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064059.htm M10334) Potassium cyanide (KCN) broth ( HYPERLI

46、NK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063514.htm M12635) Phenol red carbohydrate broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063494.htm M12) Purple carbohydrate broth ( HYPERLINK /Food

47、/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063529.htm M13037) MacConkey agar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064496.htm M9) Nutrient broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteri

48、ologicalAnalyticalManualBAM/ucm064169.htm M11439) Brain heart infusion (BHI) broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063362.htm M2440) Papain solution, 5% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalytic

49、alManualBAM/ucm063490.htm M56a41) Cellulase solution, 1% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064568.htm M18742) Tryptose blood agar base ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm06287

50、1.htm M16643) Universal preenrichment broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062927.htm M18844) Universal preenrichment broth (without ferric ammonium citrate) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAn

51、alyticalManualBAM/ucm062933.htm M188a45) Buffered peptone water ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063366.htm M19246) Dey-Engley broth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064244

52、.htm M19347) Potassium sulfite powder, anhydrous Chlorine solution, 200 ppm, containing 0.1% sodium dodecyl sulfate ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061212.htm R12a48) Ethanol, 70% ( HYPERLINK /Food/ScienceResearch/LaboratoryMeth

53、ods/BacteriologicalAnalyticalManualBAM/ucm061906.htm R2349) Kovacs reagent ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062242.htm R3850) Voges-Proskauer (VP) test reagents ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalA

54、nalyticalManualBAM/ucm061693.htm R8951) Creatine phosphate crystals Potassium hydroxide solution, 40% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062972.htm R6552) 1 N Sodium hydroxide solution ( HYPERLINK /Food/ScienceResearch/LaboratoryMe

55、thods/BacteriologicalAnalyticalManualBAM/ucm063321.htm R7353) 1 N Hydrochloric acid ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062240.htm R3654) Brilliant green dye solution, 1% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteriol

56、ogicalAnalyticalManualBAM/ucm061201.htm R855) Bromcresol purple dye solution, 0.2% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061203.htm R956) Methyl red indicator ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalytic

57、alManualBAM/ucm062248.htm R4457) Sterile distilled water Tergitol Anionic 7 ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063332.htm R7858) Triton X-100 ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/uc

58、m061652.htm R8659) Physiological saline solution, 0.85% (sterile) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062966.htm R6360) Formalinized physiological saline solution ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAn

59、alyticalManualBAM/ucm061921.htm R2761) Salmonella polyvalent somatic (O) antiserum Salmonella polyvalent flagellar (H) antiserum 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 Salmonella Spicer-Edwards flagellar (H) a

60、ntiseraPreparation of foods for isolation of SalmonellaThe following methods are based on the analysis of a 25 g analytical unit at a 1:9 sample/broth ratio. Depending on the extent of compositing, add enough broth to maintain this 1:9 ratio unless otherwise indicated. For samples not analyzed on an