Boehnke Kevin F, Eaton Kathryn A, Fontaine Clinton, Brewster Rebecca, Wu Jianfeng, Eisenberg Joseph N S, Valdivieso Manuel, Baker Laurence H, Xi Chuanwu
Department of Environmental Health Sciences, University of Michigan, Ann Arbor, MI, USA.
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, USA.
Helicobacter. 2017 Aug;22(4). doi: 10.1111/hel.12391. Epub 2017 Apr 24.
Helicobacter pylori infection has been consistently associated with lack of access to clean water and proper sanitation, but no studies have demonstrated that the transmission of viable but nonculturable (VBNC) H. pylori can occur from drinking contaminated water. In this study, we used a laboratory mouse model to test whether waterborne VBNCH. pylori could cause gastric infection.
We performed five mouse experiments to assess the infectivity of VBNCH. pylori in various exposure scenarios. VBNC viability was examined using Live/Dead staining and Biolog phenotype metabolism arrays. High doses of VBNCH. pylori in water were chosen to test the "worst-case" scenario for different periods of time. One experiment also investigated the infectious capabilities of VBNC SS1 using gavage. Further, immunocompromised mice were exposed to examine infectivity among potentially vulnerable groups. After exposure, mice were euthanized and their stomachs were examined for H. pylori infection using culture and PCR methodology.
VBNC cells were membrane intact and retained metabolic activity. Mice exposed to VBNCH. pylori via drinking water and gavage were not infected, despite the various exposure scenarios (immunocompromised, high doses) that might have permitted infection with VBNCH. pylori. The positive controls exposed to viable, culturable H. pylori did become infected.
While other studies that have used viable, culturable SS1 via gavage or drinking water exposures to successfully infect mice, in our study, waterborne VBNC SS1 failed to colonize mice under all test conditions. Future studies could examine different H. pylori strains in similar exposure scenarios to compare the relative infectivity of the VBNC vs the viable, culturable state, which would help inform future risk assessments of H. pylori in water.
幽门螺杆菌感染一直与缺乏清洁水和适当卫生设施有关,但尚无研究表明饮用受污染的水会导致活的但不可培养(VBNC)的幽门螺杆菌传播。在本研究中,我们使用实验室小鼠模型来测试水源性VBNC幽门螺杆菌是否会引起胃部感染。
我们进行了五项小鼠实验,以评估VBNC幽门螺杆菌在不同暴露情况下的感染性。使用活/死染色和Biolog表型代谢阵列检测VBNC的活力。选择高剂量的水中VBNC幽门螺杆菌在不同时间段测试“最坏情况”。一项实验还通过灌胃研究了VBNC SS1的感染能力。此外,对免疫功能低下的小鼠进行暴露,以检查潜在易感群体中的感染性。暴露后,对小鼠实施安乐死,并使用培养和PCR方法检查其胃部是否感染幽门螺杆菌。
VBNC细胞的细胞膜完整并保留代谢活性。尽管存在各种可能允许VBNC幽门螺杆菌感染的暴露情况(免疫功能低下、高剂量),但通过饮水和灌胃暴露于VBNC幽门螺杆菌的小鼠未被感染。暴露于活的、可培养的幽门螺杆菌的阳性对照确实被感染了。
虽然其他研究通过灌胃或饮水暴露使用活的、可培养的SS1成功感染了小鼠,但在我们的研究中,水源性VBNC SS1在所有测试条件下均未能在小鼠体内定殖。未来的研究可以在类似的暴露情况下检查不同的幽门螺杆菌菌株,以比较VBNC状态与活的、可培养状态的相对感染性,这将有助于为未来水中幽门螺杆菌的风险评估提供信息。