6 12 Vol. 6 No. 12 215 12 Journal of Food Safety and Quality Dec., 215 ONPG 李 建 煌 1, 姚 蕾 1, 李 玉 优 1, 李 焕 仪 1, 何 亮 辉 1, 姜 春 宝 2, 曹 维 强 (1., 51852; 2., 5166) 1* 摘 要 : 目 的 βd(onitrophenylβdgalactopyranoside, ONPG) 方 法 ONPG (lauryl sulfate tryptose, LST), χ 2 test ; ONPG LST, Wilcoxon 结 果 ONPG LST 48 h (P>.5); ONPG LST 48 h (P>.5) 结 论 ONPG 关 键 词 : ONPG; ; Rapid evaluation of coliforms from food samples in ONPG medium LI JianHuang 1, YAO Lei 1, LI YuYou 1, LI HuanYi 1, HE LiangHui 1, JIANG ChunBao 2, CAO WeiQiang 1* (1. Shenzhen Sino Assessment Group Co., Ltd., Shenzhen 51852, China; 2. Huizhou EntryExit Inspection and Quarantine Bureau, Huizhou 5166, China) ABSTRACT: Objective To explore the application of onitrophenylβdgalactopyranoside (ONPG) medium in the rapid evaluation of coliforms from food samples. Methods The detection results of Escherichia coli at different time in ONPG medium and lauryl sulfate tryptose (LST) were compared and analyzed by χ 2 test analysis. The results of coliforms form food samples in ONPG medium and LST were also compared, and analyzed by Wilcoxon analysis. Results There was no significant difference between the results of E. coli in ONPG medium (in ) and LST (in 48 h) (P>.5). There was also no significant difference between the results of coliforms from food samples in ONPG medium (in ) and LST (in 48 h) (P>.5). Conclusion ONPG medium can be used for rapid evaluation of coliforms from food samples. KEY WORDS: onitrophenylβdgalactopyranoside; coliforms; rapid evaluation 1 引 言,, (coliforms) [1],, GB 489.21, MPN (48~96 h), βd(onpg),, ONPG * 通 讯 作 者 :,,, Email: caowq@sagchina.com *Corresponding author: CAO WeiQiang, Ph.D, Senior Engineer, Shenzhen Sino Assessment Group Co., Ltd., Shenzhen 51852, China. Email: caowq@sagchina.com
484 6 [25], ONPG 2 材 料 与 方 法 2.1 标 准 菌 株 及 菌 悬 液 (ATCC 25922), 1 5 ml,.5 2.2 试 剂 及 耗 材 (LST) ; ; ONPG ; ;.45 μm (PALL) 2. 实 验 样 品, 12, 4, 55 2.4 原 理 ONPG β (ONP) [6], β, 24~48 h, ONPG (1~ h), β,,, 2.5 试 剂 配 制 ONPG : 1 g,.5125 g,.2 g, 5 ml, 121 /15 min ONPG 5 mg 5 ml, 1 ml.45 μm,, 1 ml/ 1 μl/, 4 : 8.5 g 1 L, 9 ml, 121 /15 min, 4 LST : 5.6 g() 1.2 g(), 1 L,, 1 ml/, 121 /15 min, LST () LST (), 4 2.6 测 试 方 法 2.6.1 标 准 菌 株 测 试.5 1, 1 1, 1 1, 1 1 ml/ LST () 1 μl/ ONPG, 1 ml, 1mL 1 1 ml/ LST () 1 1 μl/ ONPG, 6 ±1 1 2 4 18 24 48 h, ONPG,, ; LST,,, 2.6.2 样 品 测 试 25 g, 225 ml, 2 min, 1: 1, 1: 1 1 ml, 9 ml,, 2, 1: 1,,, LST ONPG, 1 ml, 1 ml/ LST () 1 ml/ ONPG ; 1 ml, 1 ml/ LST ( ) 1 μl/ ONPG 6 ±1 : 1: 1 μl ONPG +1 ml ; 2: 1 ml LST +1 ml, : 1 μl ONPG +1 ml, 1 2,,, 4, ONPG,,,, ; LST 48 h,,, 1 2,, MPN, MPN 2. 数 据 处 理 SPSS 19., χ 2 test, Wilcoxon
12, : ONPG 4841 结 果 与 分 析.1 对 标 准 菌 株 菌 悬 液 的 测 试 结 果 LST (), 1; ONPG, 2 表 1 LST 肉 汤 阳 性 管 数 Table 1 Positive number of LST (h) 1 2 4 18 24 48 1 1 1 1 5 1 1 1 1 1 2 表 2 ONPG 培 养 基 阳 性 管 数 Table 2 Positive number of ONPG medium 1 1 1 2 1 1 4 1 5 1 6 1 1 8 1 9 1 1 1 1 1 1 1 2 (h) 1 2 4 18 24 48 1 1 1 2 1 1 4 1 5 1 6 1 1 8 1 9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1.2 对 标 准 菌 株 菌 悬 液 测 试 的 分 析 结 果 χ 2 test, 1 5 1 6 1 1 1 LST 表 χ 2 test 分 析 结 果 P 值 Table P value of χ 2 test 24 h 48 h. 对 样 品 的 测 试 结 果 ONPG 1 5 1 6 1 <.5 <.5 <.5 >.5 >.5 LST ONPG 55 MPN 4 # 表 4 55 个 实 验 样 品 的 测 试 结 果 Table 4 Test results of 55 samples (g) LST MPN (MPN/g) ONPG MPN (MPN/g) 1. 1 46 1 46 2. >11 >11. >11 >11 4. >11 >11 5. 24 1 4. 6. 1 4. 1 4.. >11 <. 8. 1.6.2 9. >11 >11 1. <. <. 11. <. <. 12. <. <. 1. >11 >11 14. <. <. 15. >11 >11 16. >11 >11 1. >11 >11 18. >11 >11 19. >11 >11 2. <. <. 21. <. <. 22. <. <. 2. >11 >11 24. >11 >11 25. <. <. 26. >11 >11 2. <. <. 28. >11 >11 29. >11 >11. >11 >11 1. >11 >11
4842 6 # (g) LST MPN (MPN/g) ONPG MPN (MPN/g) 续 表 4 2. <. <.. >11 >11 4. <. <. 5. >11 >11 6. >11 2 11. >11 >11 8. >11 11.2 9. >11 >11 4. 2 11 2 11 41. 2 11 24 42. <. <. 4. >11 >11 44. >11 >11 45. 1 16 24 46. >11 >11 4. <. <. 48. >11 >11 49. >11 >11 5. >11 >11 51. >11 >11 52. >11 >11 5. >11 >11 54. >11 >11 55. >11 >11 # : :,, 4 讨 论 (1) ()(),,, [9],, 2~,.2 g/l,, [6] ONPG,.5~1. g/l, [1],.2 g/l ONPG.5 g/l (2), ONPG 4 h, 24 h 48 h ; ONPG LST 48 h (P>.5), ONPG, LST 48 h (P>.5) (1 5 1 6 1 ) (P<.5), 24 h 48 h,, ONPG () Wilcoxon 55 MPN, : ONPG LST (P=.116>.5), ONPG (4) ONPG,, : 1 ONPG, 4, ONPG, LST 1/1, 参 考 文 献 [1] GB 489.21 [S]. GB 489.21 National food safety standard Food microbiological examination: Enumeration of coliforms [S]. [2] Chao W. Evaluation of Colilert18 for the detection of coliforms and Escherichia coli in tropical fresh water [J]. Lett Appl Microbiol, 26, 42(2): 115 12. [] Manafi M, Kneifel W. A combined chromogenicfluorogenic medium for the simultaneous detection of total coliforms and Escherichia coli in water [J]. Zentralblatt Fur Hygiene Und Umweltmedizin, 1989, 189(): 225 24. [4]. ONPG [J]., 211, : 24 28. Zhang ZL. Comparison of ONPG single tube quantitative assay method and MPN method in the coliforms mensuration from seawater [J]. J Fujian Fish, 211.: 24 28. [5],,,.
12, : ONPG 484 [J]., 26, 5: 49 498. Sun ZK, Wu R, Ding P, et al Comparison between rapid detection method of enzyme substrate technique and multipletube fermentation technique in water coliform bacteria detection [J]. J Hyg Res, 26, 5: 49 498. [6]. [D]. :, 212. Chen CY. Establishment and Evaluation of cultural medium for rapid detection of coliform in food samples [D]. Zhengzhou: Zhengzhou University, 212. [],. [J]., 1986, 4: 5 6. Yang Y, E TS. Sodium lauryl sulfate instead bile salt for the evaluation of coliforms [J]. J Microbiol, 1986, 4: 5 6 [8],,,. [J]., 199, : 2. Yang BL, Wang SY, Wang SZ, et al. Comparison of antibacterial effect of different bile salts [J]. Chin J Food Hyg, 199, : 2 [9],,. [J]., 215, 29: 98 99. Xue H, Jiang L, Tian HG. Differences of three kinds of national standard method for the detection of coliforms in food samples [J]. Chin Primary Health Care, 215, 29: 98 99 [1],,,. [J]., 215, 8: 9 96. Wang B, Wang WD, Li PT,et al. Optimization of a marine bacterium chromogenic medium [J]. Chin Agric Sci Bull, 215, 8: 9 96. 作 者 简 介 ( 责 任 编 辑 : 白 洪 健 ) 李 建 煌, 主 要 研 究 方 向 为 食 品 微 生 物 检 测 Email: yuyugufeng@16.com 曹 维 强, 博 士, 高 级 工 程 师, 主 要 研 究 方 向 为 食 品 安 全 检 测 Email: caowq@sagchina.com