1、 KS J ISO 7937 KSKSKSKS SKSKSKS KSKSKS SKSKS KSKS SKS KS KS J ISO 7937 :2007 (2012 ) 2007 11 23 http:/www.kats.go.krKS J ISO 7937:2007 : ( ) ( ) () () () ( ) : (http:/www.standard.go.kr) : :2002 10 31 :2007 11 23 :2012 12 28 : 20120814 : ( 02-509-7270) (http:/www.kats.go.kr). 10 5 , . KS J ISO 7937:
2、2007 (2012 ) Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of Clostridium perfringens Colony-count technique 2004 3 ISO 7937, Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of Clostridium perfringensColony-count technique .
3、 ISO 7937 ISO/TC 34 (SC9) . 1 (Clostridium perfringens) . . 2 . . ( ) . KS H ISO 68871, , 10 1: 10 KS H ISO 68872, , 10 2: KS H ISO 68873, , 10 3: KS H ISO 68874, , 10 4: , , KS H ISO 7218, KS H ISO 111331, 1: KS J ISO 7937:2007 ISO 8261, Milk and milk productsGeneral guidance for the preparation of
4、 test samples, initial suspensions and decimal dilutions for microbiological examination ISO/TS 111332:2003, Microbiology of food and animal feeding stuffsGuidelines on preparation and production of culture mediaPart 2:Practical guidelines on performance testing of culture media 3 . 3.1 ( Clostridiu
5、m perfringens) ( ) 3.2 mL g 4 4.1 . 10 . (poured-plate technique) . 4.2 37 (202) . 4.3 . 4.4 mL g . 5 , , KS H ISO 7218, KS H ISO 111331 ISO/TS 11133 2 . 5.1 KS H ISO 6887 ISO 8261 . 5.2 (SC) “ (egg-yolk-free tryptose-sulfite-cycloserine agar)” ( 1 ). 2 KS J ISO 7937:2007 5.2.1 5.2.1.1 (enzymatic di
6、gest of protein) 15.0 g (enzymatic digest of soya) 5.0 g (yeast extract) 5.0 g (Na 2 S 2 O 5 ) , (anhydrous) 1.0 g (III) ammonium iron(III) citrate a1.0 g (agar) 9.0 g18.0 g b 1 000 mL a 15 %(m/m) . b . 5.2.1.2 . 25 pH (7.60.2) . . 121 15 . (53) . 2 . (9.4.3.1 ), (5.5) (5.8) SC . 15 mL (6.10) 44 47
7、. (KS H ISO 7218 ). 5.2.2 D- 5.2.2.1 D-(D-cycloserine) 4.0 ga 100 mL a . 5.2.2.2 D - . 3 2 . 4 . 5.2.3 (9.2 ) 44 47 (5.2.1) 100 mL D - (5.2.2) 1 mL . 3 KS J ISO 7937:2007 5.2.4 SC , KS H ISO 111331 . ISO/TS 111332:2003, B.1TS(C) . 5.3 (fluid thioglycollate medium) 5.3.1 (enzymatic digest of casein)
8、15.0 g L-(L-cysteine) 0.5 g D-(D-glucose) 5.5 g (yeast extract) 5.0 g (sodium chloride) 2.5 g (sodium thioglycollate)(mercaptoacetate) 0.5 g (agar) 0.5 g2.0 g a(resazurin) 0.001 g 1 000 mL a . 5.3.2 . 25 pH (7.10.2) . 10 mL 121 15 . . 5.3.3 , KS H ISO 111331 . ISO/TS 111332:2003, B.4 . 5.4 (LS) (lac
9、tose sulfite medium) () 5.4.1 5.4.1.1 (enzymatic digest of casein) 5.0 g (yeast extract) 2.5 g (sodium chloride) 2.5 g (lactose) 10 g L- (L-cysteine hydrochloride) 0.3 g 1 000 mL 4 KS J ISO 7937:2007 5.4.1.2 ( ) . 25 pH (7.10.2) . (6.7) 8 mL 121 15 . 3 2 4 . 5.4.2 (disodium disulfite) 5.4.2.1 (Na 2
10、S 2 O 5 ), (anhydrous) 1.2 g 100 mL 5.4.2.2 . 1 . 5.4.3 ()ammonium iron(III) citrate 5.4.3.1 (III) ammonium iron(III) citrate 1 g 100 mL 5.4.3.2 (III) . 1 . 5.4.4 . , . (5.4.1) 8 mL (5.4.2) 0.5 mL (III) (5.4.3) 0.5 mL . 1 . 5.5 (nitrate motility medium) () 5 KS J ISO 7937:2007 5.5.1 (enzymatic diges
11、t of casein) 5.0 g (meat extract) 3.0 g (galactose) 5.0 g (glycerol) 5.0 g (KNO 3 ) 1.0 g (Na 2 HPO 4 ) 2.5 g (agar) 1.0 g5.0 g a 1 000 mL a . 5.5.2 . 25 pH (7.30.2) . 10 mL 121 15 . , 5 3 . 15 . 4 . 5.6 (nitrite detection reagent) () 5.6.1 5-2-(5-2-ANSA) 0.1 g 5-2-ANSA 100 mL 15 %() . . 5 3 (點滴器) .
12、 . 5.6.2 0.4 g 100 mL 15 %() . . 5 3 ( .) . 5.6.3 (5.6.1 5.6.2) . . 5.7 (zinc dust) () 5.8 - (lactose-gelatin medium) () 5.8.1 6 KS J ISO 7937:2007 (enzymatic digest of casein) 15.0 g (yeast extract) 10.0 g (lactose) 10.0 g (gelatin) 120.0 g (phenol red) 0.05 g 1 000 mL 5.8.2 . 25 pH (7.50.2) . , 10
13、 mL 121 15 . , 5 3 . , 15 . 3 . 6 (KS H ISO 7218 ) . 6.1 () ( ) KS H ISO 7218 . 6.2 37 1 . 6.3 6.4 pH 25 0.01 pH , 0.1 pH . 6.5 (loops) 3 mm . , 1 6.6 . 6.7 , 35 mm 7 mm 16 mm160 mm . 7 KS J ISO 7937:2007 6.8 (total-delivery graduated pipettes) (micropipettes) 0.1 mL 0.5 mL 1 mL 10 mL 6.9 90 mm100 m
14、m . 6.10 44 47 46 0.5 6.11 ( ) . 7 . . , . 8 KS H ISO 68872, KS H ISO 68873, KS H ISO 68874 ISO 8261 . 9 9.1 , KS H ISO 6887 ISO 8261 . 9.2 ( ) , (6.9) 1 mL (6.8) . (6.10) 44 47 SC (5.2.3) 10 mL15 mL . , SC 10 mL . . (6.3) 37 (202) . . 10 (9.1 ). 9.3 (9.2 ) , 150 . , . 8 KS J ISO 7937:2007 . 5 , 9.4
15、.2 9.4.3 . 9.4 9.4.1 9.4.2 9.4.3 . KS H ISO 7218 . 9.4.2 LS 46 LS (5.4) . . 9.4.2.1 (9.3 ) (5.3) . 37 1824 . LS 5 . (6.10) 46 1824 . 9.4.2.2 LS ( ) . 1/4 . , 1/4 LS LS 5 (9.4.2.1) . 46 (6.10) 1824 . . SC LS . . 9.4.3 9.4.3.1 . (, , ), (5.3) . 37 1824 . SC (5.2.1.2 ) , 10 mL SC . 9 KS J ISO 7937:2007
16、 , 37 1824 . . , SC . 9.4.3.2, 9.4.3.3 9.4.3.4 . 9.4.3.2 (9.3 ) (5.5) . 37 24 . , . . (6.8) (6.11) 0.2 mL0.5 mL (5.6) . , (fume hood) . . 15 , (5.7) 10 . , . 9.4.3.3 (9.3 ) (5.8) . 37 24 . , ( ) . . 5 1 . , 24 . 9.4.3.4 SC , , , , 48 . (, ) . . 10 10.1 KS H ISO 7218 . 10.2 10.2.1 10 KS J ISO 7937:20
17、07 ( 2 ) . A . , . . 10.2.2 (log 10 -) (g mL ) , 5 % (r) . , (r) , . r . r LS 0.21, MN/LG 0.25(log 10 - ), r LS 1.67, MN/LG 1.8( ) ( A.4 ) . r LS 0.13, MN/LG 0.12(log 10 - ), r LS MN/LG 1.3( ) 1 g 10 000 1.010 4 . 1.9 . 1 g 5 263(= 10 000/1.9) 19 000 (10 0001.9) . 10.2.3 (log 10 -) (g mL ) , 5 % (R)
18、 . (R) , 1 . . 1)1) , .11 KS J ISO 7937:2007 1 R LS MN LG R (log) aR bR (log) aR b 0.26 1.8 0.31 2.1 0.55 3.5 0.52 3.3 0.65 4.5 0.72 5.3 0.27 1.9 0.29 1.9 aR(log) log 10 - . bR . 1 1 g 10 000 1.010 4 . 2.1 . 1 g 4 761(= 10 000/2.1) 21 000 (10 0002.1) . 2 , ( , 100 000 log 10 5 ) , R( 0.31.), 0.59 . log 10 - 0.18(0.310.59), 1.52(10 0.18 ) . log 10 5.18(log 10 5log 10 0.18) 152 000(100 0001.52) . 0.59 95 % . 0.59 . 0.59 = 2 96 . 1 64 . 1 11 . a) b) , c) , d) e) 12 KS J ISO 7937:2007 A () , , , 15 17 ( 2 ) . / . ISO 16140 , . (RIVM) 2000 1 , A.1A.4 . A.1 (