1.適用範圍：本方法適用於禽畜肉、蛋及蛋製品等中單核球增多性李斯特菌之檢驗，惟乳品除外。

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1 286 ISSN GPN SCI Expanded 藥物食品化粧品檢驗分析 SCI Expanded 1

2 286 ( ) Lester A. Mitscher (Kansas Univ., USA) ( ) Haruki Yamada (Kitasato Institute, Japan) ( ) Kuo-Hsiung Lee (Univ. of North Carolina, USA) ( ) Ray H. Liu (Univ. of Alabama, USA) ( ) I. K. Ho (Univ. of Mississippi, USA) ( ) Peter P. Fu (NCTR, FDA, USA) ( ) Chiu S. Lin (FDA, USA) ( ) Keith Chan (Univ. of Maryland, USA) ( )

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5 / % % ( Listeria monocytogenes ) (Listeria spp) L. innocual. ivanoviil. welshimeril. seeligeri L. grayi (2.5 ~ 42) (ph) (Coleslaw) ( Mexican-style soft cheese ) 5

6 (Listeriosis) ( CFU/g) 25-30% ampicillinpenicillin gentamycin ) CFU/ ± / 1mL 0.01 ml 5 10 ml 0.1 ml mm 15 mm ml99 ml90 ml () mm mm ± ± (Blender) (Stomacher) µm ( 3 mm) : () () ,000 g 0.1g 120 g 5 mg ph ph ph , (McFarland nephelometer standard units) (Durham fermentation tube) 7 20 mm mm Staphylococcus aureus ( ATCC 49444, BCRC 14980ATCC 25923, BCRC 10781) Rhodococcus equi (ATCC 6939,BCRC 12859) Listeria monocytogenes (ATCC 19111,BCRC 14845)

7 7 286 Listeria innocua (ATCC 33090,BCRC 14843) 苷 (esculin) (lithium chloride) (nalidixic acid,sodium salt) (colistin methane sulfonate) 吖 (acriflavin)95% (bromcresol purple) (rhamnose) (xylose) (crystal violet) (ammonium oxalate) O ( safranin O 30% (sulfanilic acid) N-(1- ) [N-(1-naphthyl)ethylenediamine dihydrochloride] - (-naphthol) sodium ( ammonium) moxalactam - ( -creatine) N,N,N,N - - (N,N,N,N -tetramethyl--phenylenediamine2hcl) (Gram stain solutions) (1) ( Hucker s ) ( ) A 2 g 95% 20 ml B 0.8 g 80 ml A B 24 (2) ( ) 2 g 1 g ml 5 ml 10 ml 300 ml (3) ( ) O 2.5 g 95% 100 ml 10 ml 90 ml % 30% 5 ml 45 ml ( Physiological saline solution ) 8.5 g g 500 ml 1 N ph ml ph M (ph 6.0) 17.6 g 500 ml ph % 1 g ph M 100 ml % (2% Nalidixic acid solution ) 2.0 g 100 ml g 20 g 500 ml 1 N ph Moxalactam sodium (ammonium) moxalactam 1 g ph M 100 ml (Nitrite detection reagents) A 1 g 5 N 125 ml

8 8 286 B N-(1- ) 0.25 g 5 N 200 ml (Methyl red indicator) 0.1 g 95% 300 ml 200 ml (Voges-Proskauer reagentsvp reagents) A - 5 g 100 ml B 40 g 100 ml 酶 (Oxidase reagent) N,N,N,N g 100 ml UVM (UVM broth) 腖 (proteose peptone) 腖 (tryptone) (Lab lemco powder) (yeast extract) (NaCl) 20.0 g (KH 2 PO 4 ) 1.35 g (Na 2 HPO 4 ) 12.0 g 苷 (esculin) 1.0 g 2% (nalidixic acid) 1.0 ml 吖 (acriflavin) 12.0 mg (Fraser broth) 腖 (proteose peptone) 腖 (tryptone) (Lab lemco powder) (yeast extract) (NaCl) 20.0 g (KH 2 PO 4 ) 1.35 g (Na 2 HPO 4 ) 12.0 g 苷 (esculin) 1.0 g 2% (nalidixic acid) 1.0 ml (lithium chloride) 3.0 g 10 ml mm 吖 0.1 ml (2.5 mg/ml)5% (ferric ammonium citrate solution) 0.1 ml (Modified Oxford medium, MOX) (Columbia blood agar base) 3944 g (agar) 2.0 g 苷 (esculin) 1.0 g (ferric ammonium citrate) 0.5 g (lithium chloride) 1 1% (colistin solution) 1.0 ml ph moxalactam 2 ml 12 ml ( ) (Horse or sheep blood overlay medium, HL)

9 g % ( ) ml 5 6 ml (Trypticase soy agar, TSA) (trypticase peptone) 1 腖 (phytone peptone) (NaCl) (agar) ph (Fermentation broth) 腖 (peptone) 10.0 g (Lab lemco powder) 1.0 g (NaCl) (360 mg / 20 ml 0.1 N NaOH) 1.0 ml 900 ml ph % (xylose) (mannitol) (rhamnose) 900 ml 100 ml mm 5 ml ph (25) (Brain heart infusion agar, BHI agar) (calf brain infusion) g (beef heart infusion) g 腖 (proteose peptone) 10.0 g (NaCl) (Na 2 HPO 4 ) 2.5 g (glucose) 2.0 g ph ml 1/21/ (Brain heart infusion broth, BHI broth) (calf brain infusion) g (beef heart infusion) g 腖 (proteose peptone) 10.0 g (NaCl) (Na 2 HPO 4 ) 2.5 g (glucose) 2.0 g ph

10 CAMP (CAMP test agar) 5% (trypticase soy agar) 8 ml (Trypticase soy agar with 0.6% yeast extract, TSAYE) (trypticase soy agar, dehydrated) 40.0 g (yeast extract) 6.0 g ph 7.3 ± ml (Nitrate broth) (beef extract) 3.0 g 腖 (peptone) 1.0 g 57 ml ph (Glucose broth, MR-VP broth) 腖 (proteose peptone) 7.0 g (glucose) (K 2 HPO 4 ) (agar) 1 5 ml ph (Glucose bromcresol purple broth) (glucose) 2.0 g 腖 (peptone) 10.0 g (beef extract) 3.0 g (NaCl) (bromcresol purple) 0.04 g 8 ml ph (Motility test medium, MTM) (beef extract ) 3.0 g 腖 (peptone) 10.0 g (NaCl) (agar) 4.0 g 5 ml mm () ph 7.4 ± (Nutrient agar) (beef extract) 3 g 腖 (peptone) 5 g (agar) 15 g ph 6.8 ± g UVM 225 ml

11 g UVM 225 ml ml UVM 225 ml (2~5 18 ) ( ) l g (ml) (Tween 80 ) g (ml) ml 10 ml 苷 MOX 1/2 ( ) ~ l mm l mm HL ~26 (beta-hemolysis) BHI agarbhi broth 18~25 16~18 HL MOX 苷 BHI broth 鈎 (hanging drop) (wet mount) BHI agar 鈎 24 (coccibacilli) (1) 16~24 (2) 1 5 (3) 1 (4) 95% (5) 30 (6) (7)

12 (Umbrella motility test) BHI agar 鈎 1/ mm CAMP (CAMP test) Rhodococcus equi Staphylococcus aureus BHI agar R. equi S. aureus CAMP ( ) 35 24~48 (arrow shaped hemolytical zone) S. aureus R. equi 酶 (Catalase test) 鈎 12 3% 酶 (Oxidase test) 鈎酶 10~ (VP test) 鈎 MR-VP ml A 0.6 ml B 0.2 ml (Methyl red test) MR-VP ml (Phenol red carbohydrate utilization test) 鈎 (Glucose fermentation test) 鈎 (Reduction of nitrate) 鈎 35 18~24 A B 0.5~1 ml MOX + 2. 酶 + 3. ( ) mm + 12

13 286 5.CAMP S. aureus + R. equi 酶 β () 1 ml () 9 ml UVM ( ) 35~37 24~ MOX () 0.1, 0.01, ( g ml ) ( ( MPN/g MPN/mL ) 95% 95% ( ( MPN/ MPN/ ml (g) ml (g) g ml) g ml) <

14 286 95% 95% ( ( MPN/ MPN/ ml (g) ml (g) g ml) g ml) (1) ( ) 0.1, 0.01, ( g ml) UVM MOX MPN (MPN/g MPN/mL) (2) ( ) 1, 0.1, 0.01 (g ml) (MPN/g MPN/mL) (3) ( ) 0.01, 0.001, (g ml) (MPN/g MPN/mL) Johnson, J. L Chapter 8. Isolation and identification of Listeria monocytogenes from meat, poultry and egg products. In USDA/FSIS Microbiology Laboratory Guidebook, Volume 1, 3rd ed. Revision 3, pp Dey B. P. ed., US Government Printing Office, Washington, D.C. U.S.A. 2. Andrews, W. H., June, G.A and Sherrod, P. S Appendix 2.Most Probable Number from Serial Dilution. In FDA Bacteriological Analytical Manual. 8th ed., pp.2.07~2.12. AOAC. International, Gaithersburg, MD, USA. 3. Hitchins, A. D Chapter 10. Listeria monocytogenes. In FDA Bacteriological Analytical Manual. 8th. ed., pp AOAC International, Gaithersburg, MD. U.S.A. 4. Donnelly, C. W., Brackett, R. E., Doores, S., Lee, W. H. and Lovett, J Chapter 38. Listeria. In Compendium of methods for the microbiological examination of foods. 4th. ed., pp American Public Health Association. Washington, D.C. U.S.A. 14

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行政院公報第 021 卷第 003 期衛生勞動篇 Methods of Test for Food Microorganisms- Test of Cronobacter CFU/ (Biological safe

行政院公報第 021 卷第 003 期衛生勞動篇 Methods of Test for Food Microorganisms- Test of Cronobacter CFU/ (Biological safe Methods of Test for Food Microorganisms- Test of Cronobacter 1. 2. 2.1. 100 15 15 CFU/ 2.2. 2.2.1. (Biological safety cabinet, BSC) (class II)( ) 2.2.2. 2.2.3. 2.2.4. (Blender) (Stomacher) 2.2.5. 2000