Pathogenic potential of enterococcus isolated from healthy people and wastewater

G. Pay, Daria V. Rakitina, Marina A. Pankova, Zlata E. Fedets, T. R. Maniya, A. V. Zagaynova
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Abstract

Introduction. Efficiency of wastewater treatment plants is a key for protection of common health. At the same time, all criteria for its evaluation are concerned about the overall biomass reduction rather than on pathogens that, in low amount, can still be present in the efflux. Purpose of the study. Therefore it seems important to evaluate the effect of purification procedures on the pathogenic potential of bacteria. In the current study, it is performed using Enterococcus isolates, since pathogenic strains present considerable threat for human health, causing endocarditis, infections of urogenic tract, nosocomial infections, etc. Materials and methods. PCR was used to evaluate the presence of potentially pathogenic genes in the extracted DNA. Seven genes were tested: genes of adhesion proteins (Esp, Asa1), proteins with lytic activity (cytolysine CylA, hyaluronidase hyl and gelatinase gelE), and antibiotic resistance factors (vanA, vanB). Three hundred sixty six isolates from wastewater plants of Moscow agglomeration and 168 from feces of healthy people were screened. Results. Percentage of pathogenic isolates varied in different wastewater treatment plants (from 36 to 55%), with no relation with the volumes of treated sewage and the purification scheme of the plant. Similar species were recovered from wastewater plants and feces, with E. faecium (36% and 53%, correspondingly) and E. faecalis (28% and 38%) as most abundant. E. hirae was presented in different numbers (24% и 1.2%) as well as E. casseliflavus (3% and 0,6%). E. durans, E. thailandicus, E. avium, E. mundtii were found from 2.5 to 1%, in similar amounts from both sources. Minor species E. raffinosus, E. moraviensis, E. malodatus presented with single isolates in wastewater plants, and E. canintеstini – in feces. The E. faecalis was the leader in percentage of pathogenic potential (75–80%). The most abundant pathogenic gene was gelE (30–33% from both sources) and asa1 (18–19%). CylA was found at similar levels (4,4–4,8%). Esp was found in 9% of wastewater plants isolates and in 14% from feces. Hyl was specific to isolates from wastewater plants (2,5%), and was present in all non-monor species (E. faecium, E. faecalis, E. hirae, E. durans, E. thailandicus) and at different stages of water treatment. Vancomycin resistance genes were not detected. Limitations. When studying the pathogenic potential of enterococcal isolates from wastewater treatment plants in the city of Moscow and the Moscow region and the feces of practically healthy people, two samples were compared, consisting of 366 and 168 isolates, respectively, which represents a sufficient reference sample. The sample was limited by geography, so the conclusions can be applied to wastewater treatment plants in the city of Moscow and the Moscow region, where similar treatment schemes were used. Conclusions. The data from this study suggests the pathogenic potential of bacteria from wastewater treatment plants to be a little bit more than that of isolates from feces of healthy people. The activated sludge can be a reservoir for pathogens and can bring contamination to the environment.
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从健康人和废水中分离的肠球菌的致病潜力
引言污水处理厂的效率是保护公众健康的关键。与此同时,对其进行评估的所有标准都关注整体生物量的减少,而不是污水中仍可能存在的少量病原体。 研究目的因此,评估纯化程序对细菌致病性的影响似乎很重要。本研究使用肠球菌分离物进行评估,因为致病菌株对人类健康构成相当大的威胁,可引起心内膜炎、泌尿道感染、医院感染等。 材料和方法。采用聚合酶链式反应(PCR)评估提取的 DNA 中是否存在潜在的致病基因。检测了七个基因:粘附蛋白基因(Esp、Asa1)、具有溶菌活性的蛋白(细胞溶解酶 CylA、透明质酸酶 hyl 和明胶酶 gelE)以及抗生素耐药因子(vanA、vanB)。对来自莫斯科城市群污水处理厂的 366 个分离株和来自健康人粪便的 168 个分离株进行了筛选。 结果显示在不同的污水处理厂中,病原体分离物的比例各不相同(从 36% 到 55%),与污水处理量和污水处理厂的净化方案无关。从污水处理厂和粪便中分离出的病原体种类相似,其中以粪大肠杆菌(36% 和 53%)和屎大肠杆菌(28% 和 38%)最多。E.hirae(24% 和 1.2%)以及 E. casseliflavus(3% 和 0.6%)的数量各不相同。 E.durans、E.thailandicus、E.avium、E.mundtii的含量从2.5%到1%不等,两种来源的含量相似。次要物种 E. raffinosus、E. moraviensis、E. malodatus 在废水处理厂中有单个分离物,E. canintеstini - 在粪便中。粪肠埃希氏菌的致病率最高(75-80%)。最丰富的致病基因是 gelE(两种来源均为 30-33%)和 asa1(18-19%)。CylA 的含量与之相近(4.4%-4.8%)。在 9% 的污水厂分离物和 14% 的粪便分离物中发现了 Esp。Hyl 是污水厂分离物的特异性基因(2.5%),存在于所有非特异性物种(E. faecium、E. faecalis、E. hirae、E. durans、E. thailandicus)和水处理的不同阶段。未检测到万古霉素抗性基因。 局限性。在研究莫斯科市和莫斯科地区污水处理厂中分离出的肠球菌与健康人粪便中分离出的肠球菌的致病性时,比较了两个样本,分别包含 366 个和 168 个分离物,这代表了足够的参考样本。样本受地域限制,因此结论可适用于莫斯科市和莫斯科州采用类似处理方案的污水处理厂。 结论这项研究的数据表明,污水处理厂细菌的致病性比从健康人粪便中分离出来的细菌要高一些。活性污泥可能是病原体的储藏库,会对环境造成污染。
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