Grant Irene R, Ball Hywel J, Rowe Michael T
Department of Food Science, Food Microbiology, Queen's University of Belfast, Northern Ireland, United Kingdom.
Appl Environ Microbiol. 2002 May;68(5):2428-35. doi: 10.1128/AEM.68.5.2428-2435.2002.
Over a 17-month period (March 1999 to July 2000), a total of 814 cows' milk samples, 244 bulk raw and 567 commercially pasteurized (228 whole, 179 semi-skim, and 160 skim), from 241 approved dairy processing establishments throughout the United Kingdom were tested for the presence of Mycobacterium paratuberculosis by immunomagnetic PCR (to detect all cells living and dead) and culture (to detect viable cells). Overall, M. paratuberculosis DNA was detected by immunomagnetic PCR in 19 (7.8%; 95% confidence interval, 4.3 to 10.8%) and 67 (11.8%; 95% confidence interval, 9.0 to 14.2%) of the raw and pasteurized milk samples, respectively. Confirmed M. paratuberculosis isolates were cultured from 4 (1.6%; 95% confidence interval, 0.04 to 3.1%) and 10 (1.8%; 95% confidence interval, 0.7 to 2.8%) of the raw and pasteurized milk samples, respectively, following chemical decontamination with 0.75% (wt/vol) cetylpyridinium chloride for 5 h. The 10 culture-positive pasteurized milk samples were from just 8 (3.3%) of the 241 dairy processing establishments that participated in the survey. Seven of the culture-positive pasteurized milk samples had been heat treated at 72 to 74 degrees C for 15 s; the remainder had been treated at 72 to 75 degrees C for the extended holding time of 25 s. When typed by restriction fragment length polymorphism and pulsed-field gel electrophoresis methods, some of the milk isolates were shown to be types distinct from those of laboratory strains in regular use within the testing laboratory. From information gathered at the time of milk sample collection, all indications were that pasteurization had been carried out effectively at all of the culture-positive dairies. That is, pasteurization time and temperature conditions complied with the legal minimum high-temperature, short-time process; all pasteurized milk samples tested phosphatase negative; and post-process contamination was considered unlikely to have occurred. It was concluded that viable M. paratuberculosis is occasionally present at low levels in commercially pasteurized cows' milk in the United Kingdom.
在17个月期间(1999年3月至2000年7月),对来自英国241家经批准的乳制品加工企业的总共814份牛奶样本进行了检测,其中包括244份散装生牛奶样本和567份商业巴氏杀菌牛奶样本(228份全脂牛奶、179份半脱脂牛奶和160份脱脂牛奶),以通过免疫磁珠PCR(检测所有活细胞和死细胞)和培养(检测活细胞)来检测副结核分枝杆菌的存在。总体而言,通过免疫磁珠PCR在19份(7.8%;95%置信区间为4.3%至10.8%)生牛奶样本和67份(11.8%;95%置信区间为9.0%至14.2%)巴氏杀菌牛奶样本中检测到了副结核分枝杆菌DNA。在用0.75%(重量/体积)十六烷基吡啶氯化物进行5小时化学去污后,分别从4份(1.6%;95%置信区间为0.04%至3.1%)生牛奶样本和10份(1.8%;95%置信区间为0.7%至2.8%)巴氏杀菌牛奶样本中培养出了经确认的副结核分枝杆菌分离株。这10份培养阳性的巴氏杀菌牛奶样本仅来自参与调查的241家乳制品加工企业中的8家(3.3%)。7份培养阳性的巴氏杀菌牛奶样本在72至74摄氏度下热处理了15秒;其余样本在72至75摄氏度下进行了25秒的延长保温时间处理。当通过限制性片段长度多态性和脉冲场凝胶电泳方法进行分型时,一些牛奶分离株显示出与检测实验室常规使用的实验室菌株不同的类型。根据牛奶样本采集时收集的信息,所有迹象表明所有培养阳性的乳制品厂的巴氏杀菌都有效进行。也就是说,巴氏杀菌时间和温度条件符合法定的最低高温短时工艺要求;所有检测的巴氏杀菌牛奶样本磷酸酶呈阴性;并且认为不太可能发生加工后污染。得出的结论是,在英国,商业巴氏杀菌牛奶中偶尔会存在低水平的活副结核分枝杆菌。