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用于追踪水体中动物粪便污染来源的拟杆菌属噬菌体宿主菌株的分离。

Isolation of bacteriophage host strains of Bacteroides species suitable for tracking sources of animal faecal pollution in water.

机构信息

Department of Microbiology, University of Barcelona-Institut de l'aigua, Diagonal 645, E-08028, Barcelona, Spain.

出版信息

Environ Microbiol. 2011 Jun;13(6):1622-31. doi: 10.1111/j.1462-2920.2011.02474.x. Epub 2011 Mar 28.

Abstract

Microbial source tracking (MST) methods allow the identification of specific faecal sources. The aim is to detect the sources of faecal pollution in a water body to allow targeted, efficient and cost-effective remediation efforts in the catchment. Bacteriophages infecting selected host strains of Bacteroides species are used as markers to track faecal contaminants in water. By using a suitable Bacteroides host from a given faecal origin, it is possible to specifically detect bacteriophages of this faecal origin. It can thus be used to detect specific phages of Bacteroides for MST. With this objective, we isolated several Bacteroides strains from pig, cow and poultry faeces by applying a previously optimized methodology used to isolate the host strains from humans. The isolated strains belonged to Bacteroides fragilis and Bacteroides thetaiotaomicron. These strains, like most Bacteroides species, detected phages of the Siphoviridae morphology. Using the newly isolated host strains for phage enumeration in a range of samples, we showed that these detect phages in faecal sources that coincide with their own origin (70-100% of the samples), and show no detection or very low percentages of detection of phages from other animal origins (from 0 to 20% of the samples). Only strains isolated from pig wastewater detected phages in 50% of human sewage samples. Nevertheless, those strains detecting phages from faecal origins other than their own detected fewer phages (2-3 log₁₀ pfu·100 ml⁻¹) than the phages detected by the specific strain of the same origin. On the basis of our results, we propose that faecal source tracking with phages infecting specific Bacteroides host strains is a useful method for MST. In addition, the method presented here is feasible in laboratories equipped with only basic microbiological equipment, it is more rapid and cost-effective than other procedures and it does not require highly qualified staff.

摘要

微生物源追踪 (MST) 方法可用于识别特定的粪便来源。其目的是检测水体中的粪便污染源,以便在集水区内进行有针对性、高效且具有成本效益的修复工作。噬菌体感染所选的拟杆菌属宿主菌株被用作追踪水中粪便污染物的标记物。通过使用来自特定粪便来源的合适拟杆菌属宿主,可以特异性地检测到这种粪便来源的噬菌体。因此,它可用于检测 MST 中的特定拟杆菌属噬菌体。基于这一目标,我们应用先前优化的方法从猪、牛和家禽粪便中分离了几种拟杆菌属菌株,该方法用于从人类中分离宿主菌株。分离出的菌株属于脆弱拟杆菌和普通拟杆菌。这些菌株与大多数拟杆菌属一样,检测到形态为长尾噬菌体科的噬菌体。使用新分离的宿主菌株在一系列样本中进行噬菌体计数,我们表明这些菌株可以检测与其自身来源相吻合的粪便源中的噬菌体(70-100%的样本),并且不会检测到或很少检测到来自其他动物来源的噬菌体(0-20%的样本)。只有从猪废水分离的菌株在 50%的人类污水样本中检测到噬菌体。然而,那些检测到非自身粪便来源噬菌体的菌株检测到的噬菌体较少(2-3 log₁₀ pfu·100 ml⁻¹),比相同来源的特定菌株检测到的噬菌体少。基于我们的结果,我们提出使用感染特定拟杆菌属宿主菌株的噬菌体进行粪便源追踪是一种有用的 MST 方法。此外,本文提出的方法在配备基本微生物设备的实验室中是可行的,它比其他程序更快速且具有成本效益,并且不需要高度熟练的工作人员。

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