青藏高原野生动物源致病性大肠埃希菌调查研究

Isolation of pathogenic Escherichia coli from wild mammals and birds in the Qinghai-Tibet Plateau

  • 摘要:
    目的 为调查青藏高原野生动物携带的致病性大肠埃希菌流行情况,对采集自青海的岩羊、白唇鹿和马鹿,以及兀鹫、红嘴山鸦、岩鸽、赤麻鸭等多种鸟类粪便样本,开展大肠埃希菌分离和分型鉴定。
    方法 对2023年青海采集的237份野生动物粪便样本,采用EC肉汤增菌后ECC培养基选择性分离培养的方法进行大肠埃希菌分离培养,通过PCR检测分离株携带毒力基因的情况,并对代表性分离菌株开展全基因组测序和大肠埃希菌致病型别研究。
    结果 从青藏高原野生动物粪便中分离获得169株细菌,经鉴定包含大肠埃希菌(n=60),泛菌属(n=26)、假单胞菌属(n=23)、沙雷菌属(n=16)和肠杆菌属(n=12)等。 根据大肠杆菌携带的毒力基因,将其中46个菌株归类为不同的致病型,同时携带多种致病型毒力基因即杂合型菌株为33株,占比55.00%,最常见的是两种致病型的组合(38.33%)。 通过系统进化分析,60株大肠埃希菌中B1群菌株最多,有42株,其次分别为A群(5株)、D群(5株)、B2群(4株)、E群(3株)和C群(1株)。 所有的菌株均携带fim基因,超半数菌株携带肠道外致病性大肠埃希菌相关毒力基因,kpsDaslAhlyA基因阳性率分别为28.33%、26.67%和26.67%,astAstxstx1stx2)基因阳性率均为23.33%。 eae的阳性菌株有3株,阳性率较低(5.00%)。
    结论 致病性大肠埃希菌在青藏高原野生动物岩羊、白唇鹿、马鹿和野生鸟类中广泛存在。 高原野生动物作为致病型大肠埃希菌的存储宿主,主动发现不同的致病类型和系统进化分群菌株,为大肠埃希菌致病性和流行规律提供菌株资源和科学依据。

     

    Abstract:
    Objective To investigate the prevalence of pathogenic Escherichia coli carried by wild animals in the Qinghai-Tibet Plateau. Fecal samples were collected from wild blue sheep, white-lipped deer, and red deer from Qinghai, as well as from a variety of bird faeces including the Himalayan vulture, red-billed chough, rock pigeon, and ruddy shelduck.
    Methods A total of 237 faecal samples of wild animals and birds collecting in Qinghai in 2023 were subjected to selective isolation and cultivation of Escherichia coli using the EC broth enrichment method followed by ECC agar. The virulence genes of the isolates were detected through PCR. Furthermore, representative isolates were subjected to whole-genome sequencing and Escherichia coli pathotype characterization.
    Results There were 169 strains in total were isolated from the feces of wild animals and birds in the Qinghai-Tibet Plateau, including Escherichia coli (n=60), Pantoea spp.(n=26), Pseudomonas spp.(n=23), Serratia spp.(n=16), and Enterobacter spp.(n=12). Based on the virulence genes carried by Escherichia coli, 46 strains were classified as different pathovars, with 33 strains being hybrid pathovars carrying multiple virulence genes, accounting for 55.00%, with the most common pathovar hybrids being a combination of two pathovars (38.33%). Through phylogenetic analysis, among the 60 strains of Escherichia coli, the predominant phylogroup belonged to B1 group (42/60), followed other phylogroups are A (5 strains), D (5 strains), B2 (4 strains), E (3 strains), and C (1 strain). All strains carried the gene fim, with over half of the strains carrying extraintestinal pathogenic Escherichia coli-related virulence genes, with positive rates of genes kpsD, aslA, and hlyA being 28.33%, 26.67%, and 26.67%, respectively, and positive rates of astA and stx (stx1, stx2) genes both being 23.33%. Three strains were positive for eae, with a low positivity rate (5.00%).
    Conclusion Pathogenic Escherichia coli is widely present in wild animals such as blue sheep, white-lipped deer, red deer, and wild birds in the Qinghai-Tibet Plateau. Plateau wildlife serve as reservoir hosts for pathogenic E. coli, facilitating the discovery of various pathogenic types and phylogroup strains, thereby providing bacterial resources and scientific evidence for understanding the pathogenicity and epidemiological patterns of E. coli.

     

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