AOAC Performance Tested Methods
X-Gal) 926VRB FDA-BAMAOACI Method Committee Guidelines for Validation of Qualitative and Quantitative Food Microbiological OMA y=1.04x-0.13 (yx FDABAM)(r 2 )=0.98 85 y=0.95x+0.15r 2 =0.94 FDA-BAM PTM
Principle The Sanita-kun Coliforms device is a cultural medium for the enumeration of coliform bacteria. The device consists of a non-woven fabric to which a layer of microbial nutrients is deposited in a film. The nutrient film also contains 5-bromo-4-chloro-3-indolyl--Dgalactopyranoside, or X-gal, which is hydrolyzed by-galactosidase from coliforms to produce a visible blue dye. The non-woven fabric and nutrient film are constructed on a base film. The basic device (fabric, nutrient and base film) is then inserted between an adhesive backing to stabilize the basic device, and a transparent film. The adhesive backing and the transparent top film serve to deter evaporation, and facilitate inspection of the inoculated device. Sample solutions containing bacteria are deposited on the fabric portion of the device. Then, the sample solution diffuses throughout the entire pad to dissolve and release the nutrient compound film forming a highly viscous solution. Bacteria migrate to the surface of the fabric where coliforms metabolize the nutrients and X-gal to produce a visible blue dye after incubation, while growth of non-coliforms is inhibited with selective reagents.
X- Gal 1mL + X- Gal
1mL Sanita-kun Coliforms 35 24hours 1. 2. 1mL 3. 4. 3524 5. 20/
Inclusivity Exclusivity BGLB BGLB Cedecea lapagei 1 + + Bacillus cereus 2 - - Citrobacter 1 + + Bacillus megaterium 1 - - Citrobacter diversus 1 + + Bacillus subtilis 3 - - Citrobacter freundii 2 + + Corynebacterium pyrogenes 1 - - Citrobacter sp. from a 3 + + Corynebacterium xerosis 1 - - food Enterobacter 2 + + Enterobacter gergoviae 1 - - Enterobacter cloacae 1 + + Enterococcus hirae 1 - - Enterobacter 1 + + Lactic acid bacteria isolated from 8 - - intermedium Enterobacter sp. from a 4 + + foods Lactobacillus acidophilus 1 - - food Escherichia coli 3 + + Lactobacillus delbruedkii 1 - - Escherichia sp. from a 3 + + Leuconostoc mesenteroides 1 - - food Klebsiella oxytoca 1 + + Proteus mirabilis 1 - - Klebsiella planticola 1 + + Proteus vulgaris 2 - - Klebsiella pneumoniae 2 + + Providencia alcalifaciens 1 - - Klebsiella terrigena 2 + + Pseudomonas aeruginosa 1 - - Klebsiella sp. from foods 6 + + Rahnella aquatilis 1 - - : Salmonella Enteritidis 1 - - Salmonella Typhymurium 3 - - Serratia marcescens 2 - + Serratia rubidaea 1 - + Staphylococcus aureus 1 - - Staphylococcus sp. 5 - - Streptococcus sp. 1 - - Yersinia aldovae 1 - -
FDA FDA-BAM a)significantly higher than FDA-BAM (p<0.05) b)significantly higher than FDA-BAM (p<0.01) 1.00
FDA-BAM BAM
FDA-BAM (cfu/ml) RSD Klebsiella sp. strain No2 Klebsiella oxytoca JCM 1665 Klebsiella sp. strain No53 Klebsiella sp. strain No172 Enterobacter sp. strain No8 Klebsiella pneumoniae IID 5209 Enterobacter aerogenes IFO 13534 Klebsiella sp. strain No2 Klebsiella sp. strain No171 Klebsiella sp. strain No52 Citrobacter sp. strain No202 Escherichia sp. strain No41 Klebsiella sp. strain No172 Klebsiella pneumoniae IID 5209 Klebsiella oxytoca JCM 1665 Klebsiella sp. strain No52 Klebsiella pneumoniae IID 5209 (cfu/ml) RSD FDA-BAM FDA-BAM FDA-BAM FDA-BAM 91 92 0.10 0.08 Enterobacter 149 147 0.09 0.13 49 50 0.16 0.14 aerogenes 73 73 0.07 0.1 12 8 0.25 0.38 IFO 13534 17 14 0.29 0.43 21 27 0.28 0.19 Klebsiella 194 199 0.03 0.06 11 14 0.32 0.21 pneumoniae 100 103 0.05 0.11 1 1 1 1 JCM 1662 20 21 0.11 0.35 80 58 0.1 0.07 Klebsiella sp. 108 118 0.1 0.09 43 25 0.07 0.16 strain No171 59 64 0.17 0.09 7 5 0.3 0.2 13 12 0.31 0.25 89 74 0.09 0.15 Klebsiella sp. 53 53 0.15 0.21 47 43 0.7 0.16 strain No171 32 31 0.16 0.26 8 9 0.3 0.3 6 6 0.5 0.2 97 105 0.1 0.03 Klebsiella sp. 98 100 0.04 0.07 48 50 0.19 0.14 strain No171 55 50 0.11 0.08 10 7 0.2 0.6 11 12 0.45 0.33 97 93 0.05 0.06 Escherichia 115 121 0.06 0.19 56 47 0.12 0.13 coli IID 5208 67 54 0.06 0.13 11 7 0.45 0.3 13 10 0.23 0.1 94 94 0.14 0.15 Enterobacter 114 108 0.1 0.11 51 47 0.22 0.13 aerogenes 54 47 0.17 0.11 7 11 0.3 0.4 IFO 13534 11 10 0.09 0.3 62 52 0.1 0.13 Enterobacter 65 50 0.11 0.08 37 37 0.22 0.16 sp. strain No8 33 28 0.12 0.24 11 7 0.2 0.4 11 6 0.36 0.5 67 68 0.13 0.1 Klebsiella 73 70 0.12 0.1 41 35 0.27 0.23 oxytoca JCM 41 34 0.15 0.09 5 4 0.6 0.5 1665 9 9 0.3 0.2 34 8 0.12 0.3 Blue letter:significantly higher than FDA-BAM(P<0.05) 18 1 0.17 1 Red letter:significantly higher than FDA-BAM(P<0.01) 4 0 0.8 1 60 62 0.17 0.1 28 28 0.3 0.21 5 7 0.4 0.6 64 59 0.09 0.1 36 32 0.08 0.13 7 7 0.4 0.3 67 58 0.06 0.12 36 25 0.11 0.2 7 5 0.1 0.4 84 83 0.15 0.05 50 43 0.12 0.12 9 7 0.3 0.3 120 112 0.07 0.09 59 49 0.11 0.16 11 9 0.17 0.34 53 38 0.13 0.16 32 21 0.28 0.29 5 5 0.4 0.4 108 94 0.1 0.2 55 48 0.04 0.19 9 9 0.3 0.4
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