Pagel J E, Qureshi A A, Young D M, Vlassoff L T
Appl Environ Microbiol. 1982 Apr;43(4):787-93. doi: 10.1128/aem.43.4.787-793.1982.
Four membrane filter methods fecal coliform enumeration were evaluated and compared in six laboratories based on determination of accuracy, specificity, upper counting limit, and recovery comparability. Recovery accuracy with pure cultures ranged from 89 to 100% for m-FC, mTEC (a procedure developed for thermotolerant Escherichia coli), and m-FC2 methods (the latter incorporating a 2-h, 35 degrees C resuscitation period), but was less than 60% for the MacConkey membrane broth method. These figures dropped by approximately 40 to 55% when the cultures were subjected to temperature (10 degrees C) stress. Close to 800 colonies were verified to determine specificity. False-positive colonies occurred most frequently with the m-FC2 method (18%), whereas false-negative colonies were most common on MacConkey membrane broth (26%). In counting range experiments using a variety of samples, the highest upper counting limit was 130 colonies per filter with the mTEC procedure. Recovery comparisons were based on over 130 samples including raw surface waters, raw sewage, and chlorinated and unchlorinated sewage effluents. In general, recoveries were significantly higher with the m-FC2 and mTEC methods; however, on m-FC2, growth of nontarget background organisms was also higher in most cases. Highest recoveries from chlorinated sewage effluents were obtained by the mTEC method, and the addition of a similar resuscitation period to the m-FC procedure (m-FC2) improved fecal coliform recovery from such samples. The best overall performance for fecal coliform enumeration was obtained with the mTEC method with high recovery and low levels of background colonies, good specificity and accuracy, and a high upper counting limit. This procedure also offers the advantage of enumerating E. coli within 24 h.
在六个实验室中,基于准确性、特异性、计数上限和回收率可比性的测定,对四种膜过滤法粪便大肠菌群计数方法进行了评估和比较。对于m-FC、mTEC(一种为耐热大肠杆菌开发的方法)和m-FC2方法(后者包含2小时、35摄氏度的复苏期),纯培养物的回收率在89%至100%之间,但麦康凯膜肉汤法的回收率低于60%。当培养物受到温度(10摄氏度)胁迫时,这些数字下降了约40%至55%。通过检测近800个菌落来确定特异性。m-FC2方法出现假阳性菌落的频率最高(18%),而麦康凯膜肉汤上出现假阴性菌落最为常见(26%)。在使用各种样品的计数范围实验中,mTEC方法的最高计数上限为每个滤膜130个菌落。回收率比较基于130多个样品,包括原地表水、原污水以及经过氯化和未氯化处理的污水。总体而言,m-FC2和mTEC方法的回收率显著更高;然而,在m-FC2方法中,大多数情况下非目标背景微生物的生长也更高。mTEC方法从氯化污水中获得的回收率最高,并且在m-FC程序(m-FC2)中添加类似的复苏期可提高此类样品中粪便大肠菌群的回收率。mTEC方法在粪便大肠菌群计数方面总体表现最佳,回收率高、背景菌落水平低、特异性和准确性良好且计数上限高。该方法还具有在24小时内对大肠杆菌进行计数的优势。