1、美国FDA细菌学分析手册第八版BAM沙门氏菌模板BAM: SalmonellaNOTICE:If you are looking for BAM Chapter 5: Salmonella (December Edition) that is incorporated by reference in 21 CFR Parts 16 and 118: Federal Register Final Rule1 (July 9, , 74 FR 33030): Prevention of Salmonella Enteritidis in Shell Eggs During Production,
2、Storage, and Transportation, please use theseversions of the BAMSalmonella Chapter2 (PDF, 189 Kb)andAppendix 1: Rapid Methods for Detecting Foodborne Pathogens3 (PDF, 195 Kb).The most recentEdition of BAM Chapter 5: Salmonella is available below this notice.November VersionBacteriological Analytical
3、 ManualChapter 5SalmonellaAuthors: Wallace H. Andrews and Thomas HammackRevision History:November - Addition to Section C: Preparation of foods for isolation of Salmonella: Leafy green vegetables and herbs. February Removed link to Appendix 1: Rapid Methods for Detecting Foodborne Pathogens (now arc
4、hived). December Mamey pulp method added, and Section D revised. June Eggs method revised for shell eggs and liquid whole eggs. April Frog legs method, Lactic casein, Rennet casein, Sodium caseinate and Rabbit carcass methods revised, top ears and other dog chew toys added. Removed section A.25, Mec
5、hanical shaker. October 25, Extension of the applicability of the orange juice method in section C.19 to apple juice and apple cider. 1999-DEC, -MAR, and -AUG Final revision on -NOV-14 (see the Introduction for a summary of changes).To obtain a copy of a prior version not currently posted, please co
6、ntact Thomas HammackChapter ContentsIntroduction Equipment and Materials Media and Reagents Preparation of foods for isolation of Salmonella Isolation of Salmonella Identification of Salmonella ReferencesIntroductionSeveral changes are being introduced in this edition of BAM (8th Edition). The first
7、 change involves the expanded use of Rappaport-Vassiliadis (RV) medium4 for foods with both high 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 (5, 6) and collaborative (7,
8、 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 for the analysis of all foods, except guar gum. In addition, RV medium replaces lauryl tryptose broth for use with dry active yeast. Tetrath
9、ionate (TT)5 broth continues to be used as the second 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 refrigerati
10、ng incubated preenrichments and selective enrichments 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 lysine iron agar (LIA
11、)6 slants. In the former edition (BAM-7), triple sugar iron agar (TSI)7 and LIA slants were incubated at 35C for 24 2 h and 48 2 h, respectively. Unpublished data have demonstrated that the 48 h reading of LIA slants is without diagnostic value. Of 193 LIA slants examined, all gave definitive result
12、s within 24 2 h of incubation. No significant changes altered the final test result when the slants were incubated an additional 24 h. Thus, 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 editi
13、on (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 30 min. Mercuric chloride is classified as a hazardous waste, and is expensive to dispose of according to Environmental Protection Agency guidelines. In this e
14、dition (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 (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
15、 thoroughly mixed before analysis. After mixing the egg contents, 25 g (ml) are added to 225 ml trypticase (tryptic) soy broth supplemented with 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, nonpip
16、ettable mixture results. Addition of the enzyme cellulase to the preenrichment medium, however, results in a readily pipettable mixture.A method for orange juice (pasteurized and unpasteurized) has been included due to recent orange juice-related outbreaks.The directions for picking colonies from th
17、e selective plating agars have been made more explicit to reflect the intent of the method. In the absence of typical or suspect colonies on 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% o
18、f all Salmonella cultures isolated by FDA analysts from certain foods, especially seafoods, during the past several years have been atypical.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
19、 BAM (bismuth sulfite8, Hektoen enteric9, and xylose lysine desoxycholate agars10) and three relatively new agars (EF-18, 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
20、combination of selective plating agars recommended in BAM-7 remains unchanged.A.Equipment and Materials 1.Blender and sterile blender jars (see Chapter 111) 2.Sterile, 16 oz (500 ml) wide-mouth, screw-cap jars, sterile 500 ml Erlenmeyer flasks, sterile 250 ml beakers, sterile glass or paper funnels
21、of appropriate size, and, optionally, containers of appropriate capacity to accommodate composited samples 3.Sterile, bent glass or plastic spreader rods 4.Balance, with weights; g capacity, sensitivity of 0.1 g 5.Balance, with weights; 120 g capacity, sensitivity of 5 mg 6.Incubator, 35 2 C 7.Refri
22、gerated incubator or laboratory refrigerator, 4 2C 8.Water bath, 49 1C 9.Water bath, circulating, thermostatically-controlled, 43 0.2C 10.Water bath, circulating, thermostatically-controlled,42 0.2C 11.Sterile spoons or other appropriate instruments for transferring food samples 12.Sterile culture d
23、ishes, 15 x 100 mm, glass or plastic 13.Sterile pipets, 1 ml, with 0.01 ml graduations; 5 and 10 ml, with 0.1 ml graduations 14.Inoculating needle and inoculating loop (about 3 mm id or 10 5l), nichrome, platinum-iridium, chromel wire, or sterile plastic 15.Sterile test or culture tubes, 16 x 150 mm
24、 and 20 x 150 mm; serological tubes, 10 x 75 mm or 13 x 100 mm 16.Test or culture tube racks 17.Vortex mixer 18.Sterile shears, large scissors, scalpel, and forceps 19.Lamp (for observing serological reactions) 20.Fisher or Bunsen burner 21.pH test paper (pH range 6-8) with maximum graduations of 0.
25、4 pH units per color change 22.pH meter 23.Plastic bags, 28 x 37 cm, sterile, with resealable tape. (Items 23-24 are needed in the analysis of frog legs and rabbit carcasses.) 24.Plastic beakers, 4 liter, autoclavable, for holding plastic bag during shaking and incubation. 25.Sponges, non-bactericid
26、al (Nasco cat # B01299WA), or equivalent. 26.Swabs, non-bactericidal, cotton-tipped.B.Media12 and Reagents13For preparation of media and reagents, refer to Methods 967.25-967.28 in Official Methods of Analysis (1).1.Lactose broth (M7414) 2.Nonfat dry milk (reconstituted) (M11115) 3.Selenite cystine
27、(SC) broth (M13416) 4.Tetrathionate (TT) broth (M14517) 5.Rappaport-Vassiliadis (RV) medium (M13218). NOTE: RV medium must be made from its individual ingredients. Commercial formulations are not acceptable. 6.Xylose lysine desoxycholate (XLD) agar (M17919) 7.Hektoen enteric (HE) agar (M6120) 8.Bism
28、uth sulfite (BS) agar (M1921) 9.Triple sugar iron agar (TSI) (M14922) 10.Tryptone (tryptophane) broth (M16423) 11.Trypticase (tryptic) soy broth (M15424) 12.Trypticase soy broth with ferrous sulfate (M18625) 13.Trypticase soy-tryptose broth (M16026) 14.MR-VP broth (M10427) 15.Simmons citrate agar (M
29、13828) 16.Urea broth (M17129) 17.Urea broth (rapid) (M17230) 18.Malonate broth (M9231) 19.Lysine iron agar (LIA) (Edwards and Fife) (M8932) 20.Lysine decarboxylase broth (M87)33 21.Motility test medium (semisolid) (M10334) 22.Potassium cyanide (KCN) broth (M12635) 23.Phenol red carbohydrate broth (M
30、12136) 24.Purple carbohydrate broth (M13037) 25.MacConkey agar (M9138) 26.Nutrient broth (M11439) 27.Brain heart infusion (BHI) broth (M2440) 28.Papain solution, 5% (M56a41) 29.Cellulase solution, 1% (M18742) 30.Tryptose blood agar base (M16643) 31.Universal preenrichment broth (M18844) 32.Universal
31、 preenrichment broth (without ferric ammonium citrate) (M188a45) 33.Buffered peptone water (M19246) 34.Dey-Engley broth (M19347) 35.Potassium sulfite powder, anhydrous 36.Chlorine solution, 200 ppm, containing 0.1% sodium dodecyl sulfate (R12a48) 37.Ethanol, 70% (R2349) 38.Kovacs reagent (R3850) 39.
32、Voges-Proskauer (VP) test reagents (R8951) 40.Creatine phosphate crystals 41.Potassium hydroxide solution, 40% (R6552) 42.1 N Sodium hydroxide solution (R7353) 43.1 N Hydrochloric acid (R3654) 44.Brilliant green dye solution, 1% (R855) 45.Bromcresol purple dye solution, 0.2% (R956) 46.Methyl red indicator (R4457) 47.Sterile distilled water 48.Tergitol Anionic 7 (R7858) 49.Triton X-100 (R8659)
