Food and Environmental Testing Food Safety and Quality Reference Guide Contents PART 1 QUALITY INDICATORS 1 Preparation of Dilutions Coliforms ISO Standard 1 ISO Standards 6 Total Flora Alternative Method: RAPID'E.coli 2 7 ISO Standards 1 Enterobacteriaceae Escherichia coli ISO Standards 8 ISO Standards 3 Alternative Method: RAPID'Enterobacteriaceae 9 Alternative Method: RAPID'E.coli 2 5 Alternative Method: iQ-Check 9 Enterobacteriaceae Real-Time PCR Kit PART 2 PATHOGEN TESTING 10 Salmonella spp. Listeria monocytogenes ISO Standard 10 ISO Standards 18 FDA BAM Protocol 11 FDA BAM Protocol 20 USDA FSIS MLG Protocol 11 Health Canada Protocol 20 Alternative Method: RAPID'Salmonella 12 Alternative Methods: RAPID'L.mono 21 Double Enrichment Protocol Alternative Methods: AL 22 Alternative Method: RAPID'Salmonella 12 (Agar Listeria according to Ottaviani and Agosti) Short Protocol Alternative Method: iQ-Check Listeria 23 Alternative Method: iQ-Check Salmonella II 13 monocytogenes II Real-Time PCR Kit Real-Time PCR Kit Listeria spp. Alternative Method: iQ-Check Salmonella II, 13 Alternative Method: RAPID'L.mono 25 iQ-Check S. Enteritidis, and iQ-Check S. Typhimurium Real-Time PCR Kits Alternative Method: RAPID'Listeria spp. 25 Cronobacter spp. Alternative Method: AL 26 (Agar Listeria according to Ottaviani and Agosti) ISO Standard 15 Alternative Method: iQ-Check Listeria spp. 26 FDA BAM Protocol 15 Real-Time PCR Kit Alternative Method: RAPID'Sakazakii 16 Alternative Method: iQ-Check Cronobacter spp. 16 Real-Time PCR Kit Cronobacter spp. and Salmonella spp. Alternative Method: iQ-Check Cronobacter 17 and iQ-Check Salmonella Real-Time PCR Kits or RAPID'Sakazakii and RAPID'Salmonella This booklet is provided by Bio-Rad only for the use of Bio-Rad products. It is not a contractual document. CONTENTS PART 2 PATHOGEN TESTING (Continued) Shiga Toxin–Producing Escherichia coli Vibrio spp. (including O157:H7) ISO Standard 38 Standard Method: USDA FSIS MLG Protocol 28 FDA BAM Protocol 38 Alternative Method: iQ-Check STEC 28 Alternative Method: iQ-Check Vibrio 39 VirX and SerO II Real-Time PCR Kits Real-Time PCR Kit Alternative Method: dd-Check STEC Kit 29 Bacillus cereus Alternative Method: RAPID'E.coli O157:H7 29 ISO Standard 40 Alternative Method: iQ-Check E. coli O157:H7 30 Alternative Method: RAPID'B.cereus 40 Real-Time PCR Kit Aspergillus spp. Campylobacter spp. Alternative Method: iQ-Check Aspergillus 41 ISO Standards 31 Real-Time PCR Kit USDA FSIS MLG Protocol 32 Sulfite-Reducing Bacteria Alternative Method: iQ-Check Campylobacter 33 ISO Standard 42 Real-Time PCR Kit Clostridium perfringens Alternative Method: RAPID'Campylobacter 33 ISO Standard 43 Staphylococcus aureus Shigella ISO Standards 35 ISO Standard 44 FDA BAM Protocol 36 Alternative Method: RAPID'Staph 37 PART 3 APPENDIX 45 Validations 45 Quality and Certification 47 Summary 46 Documents and Certificates 48 Preparation of Dilutions Total Flora ISO Standards ISO 6887-1:2017 ISO 4833-1:2013 Preparation of test samples, initial suspension, and Horizontal method for the enumeration of tenfold dilutions for microbiological examination microorganisms Part 1: General rules for preparation of dilutions for Part 1: Colony count at 30°C by the pour-plate microbiological examination technique m g or v ml sample Sample/initial suspension + 9 x m g or 9 x v ml Dilutions Tryptone Salt Diluent or BPW (and following dilutions) Dilution 10–1 PCA 30 ± 1°C 72 ± 3 hr (and following dilutions) Enumeration of microorganisms 1 | Food Safety and Quality Reference Guide QUALITY INDICATORS Total Flora Bacteria PCA ISO Standard Ordering Information ISO 4833-2:2013 Horizontal method for the enumeration of Catalog # Description microorganisms Buffered Peptone Water (BPW) Part 2: Colony count at 30°C by the surface-plating BPW Plus technique Ready to use 3554179 225 ml x 6 bottles 3555795 3 L x 4 bags 3555790 5 L x 2 bags Dehydrated Sample/initial suspension 3564684 500 g 3564686 5 kg Dilutions BPW Standard Ready to use 12013260 5 L x 2 bags Dehydrated 12013259 500 g 12013258 5 kg PCA Plate Count Agar (PCA) Ready to use 3554459 100 ml x 6 bottles 30 ± 1°C 3554457 200 ml x 6 bottles 72 ± 3 hr 3563989 90 mm x 20 dishes Enumeration of microorganisms Dehydrated 3564475 500 g Tryptone Salt Diluent Ready to use 3555732 225 ml x 6 bottles 3555754 9 ml x 25 tubes 3555756 90 ml x 6 bottles 3555796 3 L x 4 bags Dehydrated 3564544 500 g Food Safety and Quality Reference Guide | 2 Escherichia coli ISO Standards ISO 16649-1:2018 ISO 16649-2:2001 Horizontal method for the enumeration of Horizontal method for the enumeration of b-glucuronidase–positive E. coli b-glucuronidase–positive E. coli Part 1: Colony-count technique at 44°C using Part 2: Colony-count technique at 44°C using membranes and 5-bromo-4-chloro-3-indolyl 5-bromo-4-chloro-3-indolyl b-D-glucuronide b-D-glucuronide Sample/initial suspension Sample/initial suspension Dilutions Dilutions Minerals modified glutamate agar with membrane 2x* Room temperature Resuscitation 15 min and 37°C 4 hr ± 15 min TBX 44°C 18–24 hr Direct enumeration TBX 44°C 20–24 hr Direct enumeration * Please refer to ISO 7218:2007. 3 | Food Safety and Quality Reference Guide QUALITY INDICATORS Escherichia coli ISO Standards ISO 16649-3:2015 ISO 7251:2005 Horizontal method for the enumeration of General guidance for detection b-glucuronidase–positive E. coli and enumeration of presumptive E. coli Part 3: Most probable number technique using Most probable number technique 5-bromo-4-chloro-3-indolyl b-D-glucuronide Sample/initial suspension Sample/initial suspension Add 10 ml to each of Subsequent dilution. Tryptose lauryl sulfate 3 tubes of double Add 1 ml to each of strength minerals 3 tubes of single modified glutamate strength minerals medium modified glutamate medium Single or* Double concentration concentration 37°C 24 ± 2 hr 37°C 24–48 ± 2 hr Subculture each tube presenting EC medium yellow coloration on a dish of TBX 44°C 44°C 24–48 ± 2 hr 20–24 hr Peptone Water 44°C 48 ± 2 hr Each tube showing blue colonies on TBX Indole test reading and calculation should be considered positive Calculate the most probable number from the number of positive tubes at each dilution * For detection method only. Food Safety and Quality Reference Guide | 4 Escherichia coli E. coli Other Coliforms Total Coliforms RAPID'E.coli 2 Ordering Information Alternative Method Catalog # Description Buffered Peptone Water (BPW) RAPID' . 2 E coli BPW Plus Enumeration of E. coli Ready to use 3554179 225 ml x 6 bottles 3555795 3 L x 4 bags 3555790 5 L x 2 bags Dehydrated 3564684 500 g 3564686 5 kg Sample/initial suspension BPW Standard Ready to use 12013260 5 L x 2 bags Dilutions Dehydrated 12013259 500 g 12013258 5 kg Kovacs Reagent (for indole detection) 3555313 15 ml x 2 bottles RAPID'E.coli 2 Peptone Water (indole free) Broth Ready to use 37 or 44°C 3554180 9 ml x 25 tubes 18–24 hr Dehydrated Direct enumeration of typical colonies* 3564334 500 g RAPID'E.coli 2 Agar Ready to use 3555297 200 ml x 6 bottles 3555299 100 ml x 6 bottles Dehydrated 3564024 500 g Tryptone Bile X-Glucuronide (TBX) Agar Ready to use 3555309 100 ml x 6 bottles Dehydrated 3564035 500 g Tryptone Salt Diluent Ready to use 3555732 225 ml x 6 bottles 3555754 9 ml x 25 tubes 3555756 90 ml x 6 bottles 3555796 3 L x 4 bags * Enumeration of typical colonies for E. coli and coliforms at 37°C; E. coli at 44°C. Dehydrated 3564544 500 g 5 | Food Safety and Quality Reference Guide QUALITY INDICATORS Coliforms Coliforms VRBL ISO Standards ISO 4831:2006 ISO 4832:2006) Horizontal method for the detection and enumeration of coliforms Horizontal method for the Microbiology of food and animal feeding stuffs enumeration of coliforms Microbiology of food and MPN technique animal feeding stuffs Colony-count technique Sample Sample/initial suspension Dilutions Dilutions Tryptose lauryl sulfate 2x* VRBL Single concentration or* Double concentration 30–37°C 24 ± 2 hr 30 or 37°C 30 or 37°C 24–48 ± 2 hr 24 ± 2 hr Enumeration of characteristic BGBLB colonies and confirmation for atypical colonies 30 or 37°C 24 hr (+ 24 hr) Coliforms – Diameter ≥0.5 mm – Purple with or without a red zone of precipitated bile Reading (gas +) * For detection method only. * Please refer to ISO 7218:2007+A1:2013. Food Safety and Quality Reference Guide | 6 Coliforms E. coli Other Coliforms Total Coliforms RAPID'E.coli 2 Alternative Method Ordering Information RAPID'E.coli 2 Enumeration of coliforms Catalog # Description Brilliant Green Bile Lactose Broth (BGBLB) Ready to use 3578024 10 ml x 25 tubes Buffered Peptone Water (BPW) BPW Plus Sample/initial suspension Ready to use 3554179 225 ml x 6 bottles 3555795 3 L x 4 bags Dilutions 3555790 5 L x 2 bags Dehydrated 3564684 500 g 3564686 5 kg BPW Standard Ready to use RAPID'E.coli 2 12013260 5 L x 2 bags Dehydrated 37°C 12013259 500 g 18–24 hr 12013258 5 kg Direct enumeration of typical colonies* RAPID'E.coli 2 Agar Ready to use 3555297 200 ml x 6 bottles 3555299 100 ml x 6 bottles Dehydrated 3564024 500 g Tryptone Salt Diluent Ready to use 3555732 225 ml x 6 bottles 3555754 9 ml x 25 tubes 3555756 90 ml x 6 bottles 3555796 3 L x 4 bags Dehydrated 3564544 500 g Violet Red Bile Lactose (VRBL) Agar Ready to use 3554799 100 ml x 6 bottles Dehydrated 3564594 500 g * Enumeration of typical colonies for E.