Hill Vincent R, Polaczyk Amy L, Hahn Donghyun, Narayanan Jothikumar, Cromeans Theresa L, Roberts Jacquelin M, Amburgey James E
Centers for Disease Control and Prevention, National Center for Infectious Diseases, Division of Parasitic Diseases, 4770 Buford Highway, Mail Stop F-36, Atlanta, GA 30341-3724, USA.
Appl Environ Microbiol. 2005 Nov;71(11):6878-84. doi: 10.1128/AEM.71.11.6878-6884.2005.
The ability to simultaneously concentrate diverse microbes is an important consideration for sample collection methods that are used for emergency response and environmental monitoring when drinking water may be contaminated with an array of unknown microbes. This study focused on developing a concentration method using ultrafilters and different combinations of a chemical dispersant (sodium polyphosphate [NaPP]) and surfactants. Tap water samples were seeded with bacteriophage MS2, Escherichia coli, Enterococcus faecalis, Cryptosporidium parvum, 4.5-microm microspheres, Salmonella enterica serovar Typhimurium, Bacillus globigii endospores, and echovirus 1. Ten-liter tap water samples were concentrated to approximately 250 ml in 12 to 42 min, depending on the experimental condition. Initial experiments indicated that pretreating filters with fetal bovine serum or NaPP resulted in an increase in microbe recovery. The addition of NaPP to the tap water samples resulted in significantly higher microbe and microsphere recovery efficiencies. Backflushing of the ultrafilter was found to significantly improve recovery efficiencies. The effectiveness of backflushing was improved further with the addition of Tween 80 to the backflush solution. The ultrafiltration method developed in this study, incorporating the use of NaPP pretreatment and surfactant solution backflushing, was found to recover MS2, C. parvum, microspheres, and several bacterial species with mean recovery efficiencies of 70 to 93%. The mean recovery efficiency for echovirus 1 (49%) was the lowest of the microbes studied for this method. This research demonstrates that ultrafiltration can be effective for recovering diverse microbes simultaneously in tap water and that chemical dispersants and surfactants can be beneficial for improving microbial recovery using this technique.
当饮用水可能被一系列未知微生物污染时,对于用于应急响应和环境监测的样本采集方法而言,能够同时浓缩多种微生物是一个重要的考量因素。本研究聚焦于开发一种使用超滤器以及化学分散剂(多磷酸钠 [NaPP])和表面活性剂不同组合的浓缩方法。向自来水样本中接种了噬菌体 MS2、大肠杆菌、粪肠球菌、微小隐孢子虫、4.5 微米微球、鼠伤寒沙门氏菌、球状芽孢杆菌芽孢和埃可病毒 1。根据实验条件,10 升的自来水样本在 12 至 42 分钟内被浓缩至约 250 毫升。初步实验表明,用胎牛血清或 NaPP 对滤器进行预处理会使微生物回收率提高。向自来水样本中添加 NaPP 会使微生物和微球的回收效率显著更高。发现对超滤器进行反冲洗可显著提高回收效率。在反冲洗溶液中添加吐温 80 可进一步提高反冲洗的效果。本研究开发的超滤方法,结合使用 NaPP 预处理和表面活性剂溶液反冲洗,被发现能够回收 MS2、微小隐孢子虫、微球和几种细菌物种,平均回收效率为 70%至 93%。埃可病毒 1 的平均回收效率(49%)是该方法所研究的微生物中最低的。这项研究表明,超滤对于同时回收自来水中的多种微生物可能是有效的,并且化学分散剂和表面活性剂可能有助于使用该技术提高微生物回收率。