Bacteriological Analytical Manual BAM Chapter 5 Salmonella

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Bacteriological Analytical Manual BAM Chapter 5 Salmonella N OT I C E If you are looking for BAM Chapter 5 Salmonella December 2007 Edition that is incorporated by reference in 21 CFR P ar ts

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1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. Final Rule July 9 2009 74 FR 33030 Prevention of Salmonella Enteritidis in Shell Eggs During. Production Storage and Transportation, November 2011 Addition to Section C Preparation of foods for isolation of Salmonella Leafy green. vegetables and herbs, February 2011 Removed link to Appendix 1 Rapid Methods for Detecting Foodborne Pathogens now. 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 caseinate 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, December 1999 March 2000 and August 2000 Final revision on November 14 2000 see the. Introduction for a summary of changes, To obtain a copy of a prior version not currently posted please contact Thomas Hammack. mailto Thomas Hammack fda hhs gov,Chapter Contents.
Introduction,Equipment and Materials,Media and Reagents. Preparation of foods for isolation of Salmonella,Isolation of Salmonella. Identification of Salmonella,References,Introduction. Several changes are being introduced in this edition of BAM 8th Edition The first change involves the expanded. use of Rappaport Vassiliadis RV medium food laboratory methods bam media m132 rappaport vassiliadis. medium 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 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 Tetrathionate TT. food laboratory methods bam media m145 tetrathionate tt broth broth continues to be used as the second. selective enrichment broth However TT broth is to be incubated at 43 C for the analysis of high microbial load. foods and at 35 C for the analysis of low microbial load foods including guar gum. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 2 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. The second change involves the option of refrigerating 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 food laboratory. methods bam media m89 lysine iron agar edwards and fife slants In the former edition BAM 7 triple sugar. iron agar TSI food laboratory methods bam media m149 triple sugar iron agar tsi and LIA slants were. incubated at 35 C 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 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 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 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 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 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 nonpipettable 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. 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 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, 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 bismuth sulfite food laboratory methods bam media m19. bismuth sulfite agar wilson and blair Hektoen enteric food laboratory methods bam media m61 hektoen. enteric he agar and xylose lysine desoxycholate agars food laboratory methods bam media m179 xylose. lysine desoxycholate xld agar 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 combination of selective plating agars recommended in BAM 7 remains unchanged. A Equipment and Materials, 1 Blender and sterile blender jars see BAM Chapter 1 food laboratory methods bam food. samplingpreparation sample homogenate, 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 of appropriate size and optionally containers of appropriate. capacity to accommodate composited samples, https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 3 25.
1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 3 Sterile bent glass or plastic spreader rods, 4 Balance with weights 2000 g capacity sensitivity of 0 1 g. 5 Balance with weights 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 43 0 2 C. 10 Water bath circulating thermostatically controlled 42 0 2 C. 11 Sterile spoons or other appropriate instruments for transferring food samples. 12 Sterile culture dishes 15 100 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 16 150 mm and 20 150 mm serological tubes 10 75 mm or 13 100. 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 4 pH units per color change. 22 pH meter, 23 Plastic bags 28 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 bactericidal Nasco cat B01299WA or equivalent. 26 Swabs non bactericidal cotton tipped, B Media food laboratory methods media index bam and Reagents food laboratory. methods reagents index bam, For preparation of media and reagents refer to Methods 967 25 967 28 in Official Methods of Analysis 1.
1 Lactose broth M74 food laboratory methods bam media m74 lactose broth. 2 Nonfat dry milk reconstituted M111 food laboratory methods bam media m111 nonfat dry milk. reconstituted, 3 Selenite cystine SC broth M134 food laboratory methods bam media m134 selenite cystine. 4 Tetrathionate TT broth M145 food laboratory methods bam media m145 tetrathionate tt broth. 5 Rappaport Vassiliadis RV medium M132 food laboratory methods bam media m132 rappaport. vassiliadis medium NOTE RV medium must be made from its individual ingredients Commercial. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 4 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. formulations are not acceptable, 6 Xylose lysine desoxycholate XLD agar M179 food laboratory methods bam media m179 xylose. lysine desoxycholate xld agar, 7 Hektoen enteric HE agar M61 food laboratory methods bam media m61 hektoen enteric he. 8 Bismuth sulfite BS agar M19 food laboratory methods bam media m19 bismuth sulfite agar. wilson and blair, 9 Triple sugar iron agar TSI M149 food laboratory methods bam media m149 triple sugar iron. 10 Tryptone tryptophane broth M164 food laboratory methods bam media m164 tryptone. tryptophane broth 1, 11 Trypticase tryptic soy broth M154 food laboratory methods bam media m154 trypticase tryptic.
12 Trypticase soy tryptose broth M160 food laboratory methods bam media m160 trypticase soy. tryptose broth, 13 MR VP broth M104 food laboratory methods bam media m104 mr vp broth. 14 Simmons citrate agar M138 food laboratory methods bam media m138 simmons citrate agar. 15 Urea broth M171 food laboratory methods bam media m171 urea broth. 16 Urea broth rapid M172 food laboratory methods bam media m172 urea broth rapid. 17 Malonate broth M92 food laboratory methods bam media m92 malonate broth. 18 Lysine iron agar LIA Edwards and Fife M89 food laboratory methods bam media m89 lysine. iron agar edwards and fife, 19 Lysine decarboxylase broth M87 food laboratory methods bam media m87 lysine decarboxylase. broth falkow salmonella, 20 Motility test medium semisolid M103 food laboratory methods bam media m103 motility test. medium semisolid, 21 Potassium cyanide KCN broth M126 food laboratory methods bam media m126 potassium. cyanide kcn broth, 22 Phenol red carbohydrate broth M121 food laboratory methods bam media m121 phenol red.
carbohydrate broth, 23 Purple carbohydrate broth M130 food laboratory methods bam media m130 purple carbohydrate. fermentation broth base, 24 MacConkey agar M91 food laboratory methods bam media m91 macconkey agar. 25 Nutrient broth M114 food laboratory methods bam media m114 nutrient broth. 26 Brain heart infusion BHI broth M24 food laboratory methods bam media m24 brain heart. infusion bhi broth and agar, 27 Papain solution 5 M56a food laboratory methods bam media m56a papain solution 5. 28 Cellulase solution 1 M187 food laboratory methods bam media m187 cellulase solution. 29 Tryptose blood agar base M166 food laboratory methods bam media m166 tryptose blood agar. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 5 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 30 Universal preenrichment broth M188 food laboratory methods bam media m188 universal. preenrichment broth, 31 Universal preenrichment broth without ferric ammonium citrate M188a food laboratory. methods bam media m188a universal preenrichment broth without ferric ammonium citrate. 32 Buffered peptone water M192 food laboratory methods bam media m192 buffered peptone water. 33 Dey Engley broth M193 food laboratory methods bam media m193 dey engley broth. 34 Potassium sulfite powder anhydrous, 35 Chlorine solution 200 ppm containing 0 1 sodium dodecyl sulfate R12a food laboratory.
methods bam r12a chlorine solution, 36 Ethanol 70 R23 food laboratory methods bam r23 ethanol solution 70. 37 Kovacs reagent R38 food laboratory methods bam r38 kovacs reagent. 38 Voges Proskauer VP test reagents R89 food laboratory methods bam r89 voges proskauer vp. test reagents,39 Creatine phosphate crystals, 40 Potassium hydroxide solution 40 R65 food laboratory methods bam r65 potassium hydroxide. solution 40, 41 1 N Sodium hydroxide solution R73 food laboratory methods bam r73 1 n sodium hydroxide. 42 1 N Hydrochloric acid R36 food laboratory methods bam r36 1 n hydrochloric acid. 43 Brilliant green dye solution 1 R8 food laboratory methods bam r8 brilliant green dye solution. 44 Bromcresol purple dye solution 0 2 R9 food laboratory methods bam r9 bromcresol purple dye. solution 02, 45 Methyl red indicator R44 food laboratory methods bam r44 methyl red indicator. 46 Sterile distilled water, 47 Tergitol Anionic 7 R78 food laboratory methods bam r78 tergitol anionic 7.
48 Triton X 100 R86 food laboratory methods bam r86 triton x 100. 49 Physiological saline solution 0 85 sterile R63 food laboratory methods bam r63 physiological. saline solution 085 sterile, 50 Formalinized physiological saline solution R27 food laboratory methods bam r27 formalinized. physiological saline solution,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 M192b food laboratory methods bam media m192b modified. buffered peptone water mbpw, https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 6 25.
1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. C Preparation of foods for isolation of Salmonella. If you are looking for BAM Chapter 5 Salmonella December 2007 Edition that is incorporated by reference in 21 CFR. Parts 16 and 118 Federal Register Final Rule food eggs egg safety nal rule July 9 2009 74 FR 33030. Prevention of Salmonella Enteritidis in Shell Eggs During Production Storage and Transportation please use. these versions of the BAM Salmonella Chapter media 79991 download PDF 189 Kb and Appendix 1 Rapid. Methods for Detecting Foodborne Pathogens media 79996 download PDF 195 Kb These 2 documents are also. available as a combined le media 83330 download PDF 382 Kb. The most recent Edition of BAM Chapter 5 Salmonella continues below this notice. The 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 exact weight basis e g frog legs refer to the specific method for. instructions, 1 Dried egg yolk dried egg whites dried whole eggs liquid milk skim milk 2 fat milk. whole and buttermilk and prepared powdered mixes cake cookie doughnut biscuit. and bread infant formula and oral or tube feedings containing egg Preferably do not. thaw frozen samples before analysis If frozen sample must be tempered to obtain analytical portion. thaw suitable portion as rapidly as possible to minimize increase in number of competing organisms or. to reduce potential of injuring Salmonella organisms Thaw below 45 C for 15 min with continuous. agitation in thermostatically controlled water bath or thaw within 18 h at 2 5 C Aseptically weigh 25 g. sample into sterile wide mouth screw cap jar 500 ml or other appropriate container For. nonpowdered samples add 225 ml sterile lactose broth food laboratory methods bam media m74. lactose broth If product is powdered add about 15 ml sterile lactose broth and stir with sterile glass. rod spoon or tongue depressor to smooth suspension Add 3 additional portions of lactose broth 10. 10 and 190 ml for total of 225 ml Stir thoroughly until sample is suspended without lumps Cap jar. securely and let stand 60 5 min at room temperature Mix well by swirling and determine pH with test. paper Adjust pH if necessary to 6 8 0 2 with sterile 1 N NaOH or 1 N HCl Cap jar securely and mix. well before determining final pH Loosen jar cap about 1 4 turn and incubate 24 2 h at 35 C. Continue as in D 1 11 below, a Shell eggs 14 15 Eggs with chipped cracked or broken shells are not included in the sample. Remove any adherent material from the egg shell surface Disinfect egg surface with a solution. consisting of 3 parts of 70 alcohol ethyl or isopropyl to 1 part iodine potassium iodide. solution Prepare 70 alcohol solution either by diluting 700 ml 100 alcohol with sterile. distilled water for a final volume of 1 000 ml or by diluting 700 ml 95 alcohol with sterile. distilled water for a final volume of 950 ml Prepare iodine potassium iodide solution by. dissolving 100 g potassium iodide in 200 300 ml sterile distilled water Add 50 g iodine and heat. gently with constant mixing until the iodine is dissolved Dilute the iodine potassium iodide. solution to 1 000 ml with sterile distilled water Store iodine potassium iodide solution in an. amber glass stoppered bottle in the dark if not used immediately Prepare the disinfection. solution by adding 250 ml iodine potassium iodide solution to 750 ml 70 alcohol solution and. mix well Submerge eggs in disinfection solution for 10 seconds make sure not less than 10. seconds Remove eggs from the solution and allow to air dry Each sample shall consist of twenty. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 7 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 20 eggs for a total of fifty 50 samples per poultry house Eggs are cracked aseptically into a 4L. sterile beaker or other suitable container by gloved hands with a change of gloves between. samples Mix samples thoroughly with a sterile tool by gloved hands until yolks are completely. mixed with the albumen with a change of gloves between samples Preenrich the 20 egg sample. by adding 2 L sterile Trypticase soy broth TSB room temperature and mix well with a sterile. tool Cover securely and incubate 24 2 h at 35 C Continue as in D 1 11 below. See Salmonella Appendix food laboratory methods bam appendix salmonella for validation. b Liquid whole eggs homogenized Combine fifteen 15 25 ml test portions into a 375 ml. composite contained in a 6 liter Erlenmeyer flask Composites are held at room temperature 20. 24 C for 96 2 h After 96 2 h add 3 375 ml sterile TSB supplemented with ferrous sulfate. food laboratory methods bam media m186 trypicase tryptic soy broth ferrous sulfate as. described above and mix well by swirling Let stand 60 5 min at room temperature Mix well by. swirling and determine pH with test paper Adjust pH if necessary to 6 8 0 2 Incubate 24 2 h. at 35 C Continue as in D 1 11 below, c Hard boiled eggs chicken duck and others If the egg shells are still intact disinfect the. shells as described above and aseptically separate the shells from the eggs Pulverize the eggs egg. yolk solids and egg white solids aseptically and weigh 25 g into a sterile 500 ml Erlenmeyer flask. or other appropriate container Add 225 ml TSB food laboratory methods bam media m154. trypticase tryptic soy broth without ferrous sulfate and mix well by swirling Continue as. described above,3 Nonfat dry milk, a Instant Aseptically weigh 25 g sample into sterile beaker 250 ml or other appropriate. container Using sterile glass or paper funnel made with tape to withstand autoclaving pour 25. g analytical unit gently and slowly over surface of 225 ml brilliant green water contained in sterile. 500 ml Erlenmeyer flask or other appropriate container Alternatively 25 g analytical units may. be composited and poured over the surface of proportionately larger volumes of brilliant green. water Prepare brilliant green water by adding 2 ml 1 brilliant green dye solution. food laboratory methods bam r8 brilliant green dye solution 1 per 1000 ml sterile distilled. water Let container stand undisturbed for 60 5 min Incubate loosely capped container without. mixing or pH adjustment for 24 2 h at 35 C Continue as in D 1 11 below. b Non Instant Examine as described for instant nonfat dry milk except that the 25 g analytical. units may not be composited, 4 Dry whole milk Examine as described for instant nonfat dry milk except that the 25 g analytical.
units may not be composited, a Lactic casein Aseptically weigh 25 g sample into sterile beaker 250 ml or other appropriate. container Using sterile glass or paper funnel made with tape to withstand autoclaving pour 25. g analytical unit gently and slowly over the surface of 225 ml Universal Preenrichment broth. contained in sterile 500 ml Erlenmeyer flask or other appropriate container Analytical units 25. g may be composited Let container stand undisturbed 60 5 min Incubate loosely capped. container without mixing or pH adjustment for 24 2 h at 35 C Continue as in D 1 11 below. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 8 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. b Rennet casein Aseptically weigh 25 g sample into sterile beaker 250 ml or other appropriate. container Using sterile glass or paper funnel made with tape to withstand autoclaving pour 25. g analytical unit gently and slowly over the surface of 225 ml lactose broth contained in sterile 500. ml Erlenmeyer flask or other appropriate container Analytical units 25 g may be composited. Let container stand undisturbed 60 5 min Incubate loosely capped container without mixing or. pH adjustment for 24 2 h at 35 C Continue as in D 1 11 below. c Sodium caseinate Aseptically weigh 25 g sample into sterile wide mouth screw cap jar 500. ml or other appropriate container Add 225 ml sterile lactose broth and mix well Analytical units. may be composited Let stand 60 min at room temperature with jar securely capped Mix well by. swirling and determine pH with test paper Adjust pH if necessary to 6 8 0 2 Loosen jar about. 1 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11 below. 6 Soy flour Examine as described for rennet casein except 25 g analytical units 25 g may not be. composited, 7 Fresh frozen or dried products products not listed elsewhere in Section C Preferably. do not thaw frozen samples before analysis If frozen sample must be tempered to obtain analytical. portion thaw below 45 C for 15 min with continuous agitation in thermostatically controlled water. bath or thaw within 18 h at 2 5 C, a Egg containing products noodles egg rolls macaroni spaghetti cheese dough. prepared salads ham egg chicken tuna turkey fruits nut meats crustaceans. shrimp crab crayfish langostinos lobster and fish Aseptically weigh 25 g sample. into sterile blending container Add 225 ml sterile lactose broth food laboratory methods bam. media m74 lactose broth and blend 2 min Aseptically transfer homogenized mixture to sterile. wide mouth screw cap jar 500 ml or other appropriate container and let stand 60 5 min at. room temperature with jar securely capped Mix well by swirling and determine pH with test. paper Adjust pH if necessary to 6 8 0 2 Mix well and loosen jar cap about 1 4 turn Incubate. 24 2 h at 35 C Continue as in D 1 11 below, b Vegetables Aseptically weigh 25 g sample into a sterile wide mouth Erlenmeyer flask or other. appropriate container Add 225 mL Universal Preenrichment UP broth M188 and mix well by. swirling Incubate at 35 2 0 C for 24 2 0 hours and continue as in D 1 11 below. 8 Dried yeast active and inactive yeast Aseptically weigh 25 g sample into sterile wide mouth. screw cap jar 500 ml or other appropriate container Add 225 ml sterile trypticase soy broth. food laboratory methods bam media m154 trypticase tryptic soy broth Mix well to form smooth. suspension Let stand 60 5 min at room temperature with jar securely capped Mix well by swirling. and determine pH with test paper Adjust pH if necessary to 6 8 0 2 mixing well before determining. final pH Loosen jar cap 1 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11 below. 9 Frosting and topping mixes Aseptically weigh 25 g sample into sterile wide mouth screw cap jar. 500 ml or other appropriate container Add 225 ml nutrient broth food laboratory methods bam. media m114 nutrient broth and mix well Cap jar securely and let stand 60 5 min at room. temperature Mix well by swirling and determine pH with test paper Adjust pH if necessary to 6 8. 0 2 Loosen jar cap about 1 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11 below. a Black pepper white pepper celery seed or flakes chili powder cumin paprika. parsley flakes rosemary sesame seed thyme and vegetable flakes Aseptically weigh. 25 g sample into sterile wide mouth screw cap jar 500 ml or other appropriate container Add. 225 ml sterile trypticase soy broth TSB food laboratory methods bam media m154. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 9 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. trypticase tryptic soy broth and mix well Cap jar securely and let stand 60 5 min at room. temperature Mix well by swirling and determine pH with test paper Adjust pH if necessary to. 6 8 0 2 Loosen jar cap about l 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11. b Onion flakes onion powder garlic flakes Aseptically weigh 25 g sample into sterile wide. mouth screw cap jar 500 ml or other appropriate container Preenrich sample in TSB. food laboratory methods bam media m154 trypticase tryptic soy broth with added K2SO3 5. g K2SO3 per 1000 ml TSB resulting in final 0 5 K2SO3 concentration Add K2SO3 to broth. before autoclaving 225 ml volumes in 500 ml Erlenmeyer flasks at 121 C for 15 min After. autoclaving aseptically determine and if necessary adjust final volume to 225 ml Add 225 ml. sterile TSB with added K2SO3 to sample and mix well Continue as in C 10a. c Allspice cinnamon cloves and oregano At this time there are no known methods for. neutralizing the toxicity of these 4 spices Dilute them beyond their toxic levels to examine them. Examine allspice cinnamon and oregano at 1 100 sample broth ratio and cloves at 1 1000. sample broth ratio Examine leafy condiments at sample broth ratio greater than 1 10 because of. physical difficulties encountered by absorption of broth by dehydrated product. For oregano cinnamon and allspice mix each sample thoroughly take 25 0 g from each of 15. subsamples for a total of 375 0 g Thoroughly mix this composite Then from the 375 0 g. composite measure 37 5 g and add to 3712 5 ml of sterile pre enrichment broth Shake and mix. well for pre enrichment For food categories requiring 30 subsamples create 2 sets of composites. each consisting of 15 subsamples for those requiring 60 subsamples create 4 sets of composites. each consisting of 15 subsamples Then follow the procedures for 15 subsamples described above. For cloves create a 375 0 g composite from 15 subsamples according to the procedures described. above Then from the 375 0 g composite measure 3 75 g and add to 3746 25 ml of sterile pre. enrichment broth Shake and mix well for pre enrichment A minimum of 37 5 g sample size is. required 10 tests should be conducted from the same 375 0 g composite For food categories. requiring 30 subsamples create 2 sets of composites each consisting of 15 subsamples for those. requiring 60 subsamples create 4 sets of composites each consisting of 15 subsamples Then. follow the procedures for 15 subsamples described above. After the pre enrichment procedures described above examine these spices as described in C 10a. above maintaining recommended sample broth ratios, 11 Candy and candy coating including chocolate Aseptically weigh 25 g sample into sterile.
blending container Add 225 ml sterile reconstituted nonfat dry milk food laboratory methods bam. media m111 nonfat dry milk reconstituted and blend 2 min Aseptically transfer homogenized mixture. to sterile wide mouth screw cap jar 500 ml or other appropriate container and let stand 60 5 min. at room temperature with jar securely capped Mix well by swirling and determine pH with test paper. Adjust pH if necessary to 6 8 0 2 Add 0 45 ml 1 aqueous brilliant green dye solution and mix well. Loosen jar caps 1 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11 below. 12 Coconut Aseptically weigh 25 g sample into sterile wide mouth screw cap jar 500 ml or other. appropriate container Add 225 ml sterile lactose broth food laboratory methods bam media m74. lactose broth shake well and let stand 60 5 min at room temperature with jar securely capped Mix. well by swirling and determine pH with test paper Adjust pH if necessary to 6 8 0 2 Add up to 2 25. ml steamed 15 min Tergitol Anionic 7 food laboratory methods bam r78 tergitol anionic 7 and. mix well Alternatively use steamed 15 min Triton X 100 food laboratory methods bam r86 triton. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 10 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. x 100 Limit use of these surfactants to minimum quantity needed to initiate foaming For Triton X. 100 this quantity may be as little as 2 or 3 drops Loosen jar cap about l 4 turn and incubate 24 2 h at. 35 C Continue as in D 1 11 below, 13 Food dyes and food coloring substances For dyes with pH 6 0 or above 10 aqueous. suspension use method described for dried whole eggs C l above For laked dyes or dyes with pH. below 6 0 aseptically weigh 25 g sample into sterile wide mouth screw cap jar 500 ml or other. appropriate container Add 225 ml tetrathionate broth food laboratory methods bam media m145. tetrathionate tt broth without brilliant green dye Mix well and let stand 60 5 min at room. temperature with jar securely capped Using pH meter adjust pH to 6 8 0 2 Add 2 25 ml 0 1. brilliant green dye solution food laboratory methods bam r8 brilliant green dye solution 1 and mix. thoroughly by swirling Loosen jar cap about 1 4 turn and incubate 24 2 h at 35 C Continue as in D. 3 11 below, 14 Gelatin Aseptically weigh 25 g sample into sterile wide mouth screw cap jar 500 ml or other. appropriate container Add 225 ml sterile lactose broth food laboratory methods bam media m74. lactose broth and 5 ml 5 aqueous papain solution food laboratory methods bam media m56a. papain solution 5 and mix well Cap jar securely and incubate at 35 C for 60 5 min Mix well by. swirling and determine pH with test paper Adjust pH if necessary to 6 8 0 2 Loosen jar cap about. 1 4 turn and incubate 24 2 h at 35 C Continue as in D 1 11 below. 15 Meats meat substitutes meat by products animal substances glandular products and. meals fish meat bone Aseptically weigh 25 g sample into sterile blending container Add 225 ml. sterile lactose broth food laboratory methods bam media m74 lactose broth and blend 2 min. Aseptically transfer homogenized mixture to sterile wide mouth screw cap jar 500 ml or other. appropriate container and let stand 60 5 min at room temperature with jar securely capped If. mixture is powder or is ground or comminuted blending may be omitted For samples that do not. require blending add lactose broth and mix thoroughly let stand for 60 5 min at room temperature. with jar securely capped, Mix well by swirling and determine pH with test paper Adjust pH if necessary to 6 8 0 2 Add up to. 2 25 ml steamed 15 min Tergitol Anionic 7 and mix well Alternatively use steamed 15 min Triton X. 100 Limit use of these surfactants to minimum quantity needed to initiate foaming Actual quantity. will depend on composition of test material Surfactants will not be needed in analysis of powdered. glandular products Loosen jar caps 1 4 turn and incubate sample mixtures 24 2 h at 35 C Continue. as in D 1 11 below, 16 Frog legs This method is used for all domestic and imported frog legs Place 15 pairs of frog legs into. sterile plastic bag and cover with sterile lactose broth at a 1 9 sample to broth g ml ratio see A 23. 24 above If single legs are estimated to average 25 g or more examine only one leg of each of 15 pairs. Place bag in large plastic beaker or other suitable container Mix well and let stand 60 5 min at room. temperature Mix well by swirling and determine pH with test paper Adjust pH if necessary to 6 8. 0 2 Place plastic bag containing the frog legs and lactose broth into plastic beaker or other suitable. container Incubate 24 2 h at 35 C Continue examination as in D 1 11 below. 17 Rabbit carcasses This method is used for all domestic and imported rabbit carcasses Place rabbit. carcass into sterile plastic bag Place bag in beaker or other suitable container Add sterile lactose broth. at a 1 9 sample to broth g ml ratio to cover carcass see A 23 24 above Mix well by swirling and let. stand 60 5 min at room temperature Mix well by swirling and determine pH with test paper Adjust. pH if necessary to 6 8 0 2 Incubate 24 2 h at 35 C Continue examination as in D 1 11 below. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 11 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 18 Guar gum Aseptically weigh 25 g sample into sterile beaker 250 ml or other appropriate container. Prepare a 1 0 cellulase solution add 1 g cellulase to 99 ml sterile distilled water Dispense into 150 ml. bottles Cellulase solution may be stored at 2 5 C for up to 2 weeks Add 225 ml sterile lactose broth. food laboratory methods bam media m74 lactose broth and 2 25 ml sterile 1 cellulase solution to. sterile wide mouth screw cap jar 500 ml or other appropriate container While vigorously stirring. the cellulase lactose broth with magnetic stirrer pour 25 g analytical unit quickly through sterile glass. funnel into the cellulase lactose broth Cap jar securely and let stand 60 5 min at room temperature. Incubate loosely capped container without pH adjustment for 24 2 h at 35 C Continue as in D 1 11. 19 Orange juice pasteurized and unpasteurized apple cider pasteurized and. unpasteurized and apple juice pasteurized Aseptically add 25 ml sample to 225 ml Universal. preenrichment broth food laboratory methods bam media m188 universal preenrichment broth in. a sterile wide mouth screw capped jar 500 ml or other appropriate container Swirl the flask. contents thoroughly Cap jar securely and let stand 60 5 min at room temperature Do not adjust pH. Incubate loosely capped container for 24 2 h at 35 C Continue as in D 1 11 below treat as a low. microbial load food, 20 Pig ears and other types of dog chew pieces Place 1 piece or 2 3 pieces if smaller sizes from.
each sample unit into sterile plastic bag Place bag into large beaker or other suitable container Add. sterile lactose broth at a 1 9 sample to broth g ml ratio to cover pieces see A 23 24 above Mix well. by swirling and let stand 60 5 min at room temperature Mix well by swirling and determine pH with. test paper Adjust pH if necessary to 6 8 0 2 Add either steamed 15 min Tergitol Anionic 7 or. steamed 15 min Triton X 100 up to a 1 concentration For example if 225 ml lactose broth is added. the maximum volume of added surfactant is 2 25 ml Limit use of these surfactants to minimum. quantity to initiate foaming Incubate 24 2 h at 35 C Continue examination as in D 1 11 below. 21 Cantaloupes Preferably do not thaw frozen samples before analysis If frozen sample must be. tempered to obtain analytical portion thaw below 45 C for 15 min with continuous agitation in. thermostatically controlled water bath or thaw within 18 h at 2 5 C. For comminuted or cut fruit aseptically weigh 25 g sample into sterile blending container Add 225 ml. sterile Universal preenrichment broth food laboratory methods bam media m188 universal. preenrichment broth UP and blend 2 min Aseptically transfer homogenized mixture to sterile wide. mouth screw cap jar 500 ml or other appropriate container and let stand 60 5 min at room. temperature with jar securely capped Do not adjust pH Mix well and loosen jar cap about 1 4 turn. Incubate 24 2 h at 35 C Continue as in D 1 11 below. For whole cantaloupes do not rinse even if there is visible dirt Examine the cantaloupes as is. Place the cantaloupe into a sterile plastic bag Add enough UP food laboratory methods bam media. m188 universal preenrichment broth broth to allow the cantaloupe to float The volume of UP. food laboratory methods bam media m188 universal preenrichment broth broth may be 1 5 times. the weight of the cantaloupes For example cantaloupes weighing 1500 g will probably need a volume. of approximately 2250 ml UP food laboratory methods bam media m188 universal preenrichment. broth broth to float Add more broth if necessary Place the plastic bag with cantaloupes and UP. food laboratory methods bam media m188 universal preenrichment broth broth into a 5 liter. beaker or other appropriate container for support during incubation Allow the open end flap of the. plastic bag to fold over so as to form a secure but not air tight closure during incubation. Let stand for 60 5 min at room temperature Do not adjust pH Incubate slightly opened bag. containing cantaloupe for 24 2 h at 35 C Continue as in D 1 11 below. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 12 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 22 Mangoes Preferably do not thaw frozen samples before analysis If frozen sample must be tempered. to obtain analytical portion thaw below 45 C for 15 min with continuous agitation in thermostatically. controlled water bath or thaw within 18 h at 2 5 C. For comminuted or cut fruit aseptically weigh 25 g sample into sterile blending container Add 225 ml. sterile buffered peptone water BPW food laboratory methods bam media m192 buffered peptone. water bpw and blend 2 min Aseptically transfer homogenized mixture to sterile wide mouth screw. cap jar 500 ml or other appropriate container and let stand 60 5 min at room temperature with jar. securely capped Mix well by swirling and determine pH with test paper Adjust pH if necessary to 6 8. 0 2 Mix well and loosen jar cap about 1 4 turn Incubate 24 2 h at 35 C Continue as in D 1 11. For whole mangoes do not rinse even if there is visible dirt Examine the mangoes as is. Place the mango into a sterile plastic bag Add enough BPW food laboratory methods bam media. m192 buffered peptone water bpw to allow the mango to float The volume of BPW food laboratory. methods bam media m192 buffered peptone water bpw may be 1 0 times the weight of the mangoes. For example mangoes weighing 500 g will probably need a volume of approximately 500 ml BPW. food laboratory methods bam media m192 buffered peptone water bpw broth to float Add more. broth if necessary Place the plastic bag with mangoes and BPW food laboratory methods bam. media m192 buffered peptone water bpw broth into a 5 liter beaker or other appropriate container. for support during incubation, Let stand for 60 5 min at room temperature Adjust pH to 6 8 0 2 if necessary Incubate slightly. opened bag for 24 2 h at 35 C Continue as in D 1 11 below. 23 Tomatoes For comminuted or cut fruit aseptically weigh 25 g sample into sterile blending container. Add 225 ml Universal Preenrichment UP broth and blend 2 min Aseptically transfer homogenized. mixture to sterile wide mouth screw cap jar 500 ml or other appropriate container and let stand 60. 5 min at room temperature with jar securely capped Mix well by swirling and determine pH with test. paper Adjust pH if necessary to 6 8 0 2 Mix well and loosen jar cap about 1 4 turn Incubate 24 2. h at 35 C Continue as in D 1 11 below, For whole tomatoes do not rinse even if there is visible dirt Examine the tomatoes as is. Place the tomato into a sterile plastic bag or other suitable container sterile foil covered beaker can be. used Add enough UP food laboratory methods bam media m188 universal preenrichment broth. broth to allow the tomato to float The volume of UP food laboratory methods bam media m188. universal preenrichment broth broth may be 1 0 times the weight of the tomato For example. tomatoes weighing 300 g will probably need a volume of approximately 300 ml UP food laboratory. methods bam media m188 universal preenrichment broth broth to float Add more if necessary. Place the plastic bag if used with tomato and UP food laboratory methods bam media m188. universal preenrichment broth broth into a sterile beaker beaker size is dependent on the size of the. tomato or other appropriate container for support during incubation Allow the open end flap of the. plastic bag to fold over so as to form a secure but not air tight closure during incubation. Let stand for 60 5 min at room temperature Do not adjust pH Incubate slightly opened bag for 24. 2 h at 35 C Continue as in D 1 11 below, 24 Environmental testing Sample environmental surfaces with sterile swabs or sponges Place the. swab sponge in a sterile Whirl pak bag or equivalent that contains enough Dey Engley DE broth. food laboratory methods bam media m193 dey engley broth to cover the swab sponge. https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 13 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. Transport swabs sponges in an insulated transport container with frozen gel packs to keep the samples. cold but not frozen If samples cannot be processed immediately refrigerate at 4 2 C Start sample. analysis within 48 2 h of collection, Add swab sponge to 225 ml lactose broth in a sterile wide mouth screw capped jar 500 ml or other. appropriate container Swirl the flask contents thoroughly Cap jar securely and let stand 60 5 min at. room temperature Mix well by swirling and determine pH with test paper Adjust pH if necessary to. 6 8 0 2 Incubate 24 2 h at 35 C Continue examination as in D 1 11 below. 25 Alfalfa seeds and mung beans Aseptically weigh 25g alfalfa seeds or mung beans into a sterile 500. mL Erlenmeyer flask Aseptically add 225 mL lactose broth to the test portion and swirl the Erlenmeyer. flask Cover the mouth of the Erlenmeyer flask with sterile aluminum foil and allow contents to stand at. room temperature for 60 5 min Adjust the pH of the culture to 6 8 0 2 if necessary Incubate for. 24 2h at 35 2 C Continue as in D 1 11 below treat as high microbial load food. 26 Mamey pulp If frozen sample must be tempered to obtain analytical portion Thaw below 45 C for. 15 min with continuous agitation in thermostatically controlled water bath or thaw within 18 h at 2. For mamey pulp suspected to be contaminated with S Typhi aseptically weigh 25 g sample into sterile. wide mouth screw cap jar 500 ml or other appropriate container Add 225 ml sterile Universal. Preenrichment broth without ferric ammonium citrate food laboratory methods bam media m188a. universal preenrichment broth without ferric ammonium citrate mix by swirling and let stand 60. 5 min at room temperature with jar securely capped Do not adjust pH Mix well and loosen jar cap. about 1 4 turn Incubate 24 2 h at 35 C Continue as in D 1 11 below Treat as a low microbial load. For mamey pulp NOT suspected to be contaminated with S Typhi aseptically weigh 25 g sample into. sterile wide mouth screw cap jar 500 ml or other appropriate container Add 225 ml sterile. Universal Preenrichment broth mix by swirling and let stand 60 5 min at room temperature with jar. securely capped Do not adjust pH Mix well and loosen jar cap about 1 4 turn Incubate 24 2 h at. 35 C Continue as in D 1 11 below, 27 Fresh leafy green vegetables herbs and sprouts baby spinach cabbage iceberg lettuce.
Romaine lettuce Spring mix cilantro curly parsley culantro Italian parsley alfalfa. mung bean clover radish and broccoli sprouts Aseptically weigh 25 g into a sterile wide. mouth Erlenmeyer flask or other appropriate container Add 225 mL Universal Preenrichment UP. broth M188 for cabbage adding 225 ml modified buffered peptone water M192b and manually. mix contents by vigorously swirling the flask 25 times clockwise and 25 times counterclockwise. Incubate at 35 2 0 C for 24 2 0 hours and continue as in D 1 11 below. D Isolation of Salmonella,1 Tighten lid and gently shake incubated sample. Guar gum and foods suspected to be contaminated with S Typhi Transfer 1 ml mixture to 10. ml selenite cystine SC broth food laboratory methods bam media m134 selenite cystine broth and. another 1 ml mixture to 10 ml TT broth food laboratory methods bam media m145 tetrathionate tt. broth Vortex, All other foods Transfer 0 1 ml mixture to 10 ml Rappaport Vassiliadis RV medium. food laboratory methods bam media m132 rappaport vassiliadis medium and another 1 ml. mixture to 10 ml tetrathionate TT broth food laboratory methods bam media m145 tetrathionate. tt broth Vortex, https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 14 25. 1 21 2020 Bacteriological Analytical Manual BAM Chapter 5 Salmonella FDA. 2 Incubate selective enrichment media as follows, Foods with a high microbial load Incubate RV medium 24 2 h at 42 0 2 C circulating. thermostatically controlled water bath Incubate TT broth 24 2 h at 43 0 2 C circulating. thermostatically controlled water bath, Foods with a low microbial load except guar gum and foods suspected to be.
contaminated with S Typhi Incubate RV medium 24 2 h at 42 0 2 C circulating. thermostatically controlled water bath Incubate TT broth 24 2 h at 35 2 0 C. Guar gum and foods suspected to be contaminated with S Typhi Incubate SC and TT broths. 24 2 h at 35 C, 3 Mix vortex if tube and streak 3 mm loopful 10 l incubated TT broth on bismuth sulfite BS agar. food laboratory methods bam media m19 bismuth sulfite agar wilson and blair xylose lysine. desoxycholate XLD agar food laboratory methods bam media m179 xylose lysine desoxycholate. xld agar and Hektoen enteric HE agar food laboratory methods bam media m61 hektoen. enteric he agar Prepare BS plates the day before streaking and store in dark at room. temperature until streaked, 4 Repeat with 3 mm loopful 10 l of RV medium for samples of high and low microbial load foods and. of SC broth for guar gum, 5 Refer to 994 04 in Official Methods of Analysis 1 for option of refrigerating incubated sample. preenrichments and incubated sample selective enrichments SC and TT broths only of low moisture. foods This option allows sample analyses to be initiated as late as Thursday while still avoiding. weekend work,6 Incubate plates 24 2 h at 35 C, 7 Examine plates for presence of colonies that may be Salmonella. TYPICAL Salmonella COLONY MORPHOLOGY, Pick 2 or more colonies of Salmonella from each selective agar plate after 24 2 h incubation Typical.
Salmonella colonies are as follows, If typical colonies are present on the BS agar after 24 2 h incubation then pick 2 or more colonies. Irrespective of whether or not BS agar plates are picked at 24 2 h reincubate BS agar plates an. additional 24 2 h After 48 2 h incubation pick 2 or more typical colonies if present from the BS. agar plates only if colonies picked from the BS agar plates incubated for 24 2 h give atypical reactions. in triple sugar iron agar TSI and lysine iron agar LIA that result in culture being discarded as not. being Salmonella See sections D 9 and D 10 below for details in interpreting TSI and LIA reactions. ATYPICAL Salmonella COLONY MORPHOLOGY, In the absence of typical or suspicious Salmonella colonies search for atypical Salmonella colonies as. SUGGESTED CONTROL CULTURES, In addition to the positive control cultures typical Salmonella 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 ATCC 12325 and a lactose negative. H2S negative S abortus equi ATCC 9842 OR a lactose positive H2S negative S diarizonae ATCC. 29934 These cultures may be obtained from the American Type Culture Collection. http www atcc org http www fda gov about fda website policies website disclaimer 10801. University Boulevard Manassas VA 20110 2209, https www fda gov food laboratory methods food bacteriological analytical manual bam chapter 5 salmonella 15 25.

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