Co-infections of Klebsiella pneumoniae and Elizabethkingia miricola in black-spotted frogs (Pelophylax nigromaculatus)

文献类型： 外文期刊

第一作者： Li, Shurui

作者： Li, Shurui;Wang, Xiaoni;Lu, Yulan;Wang, Jiahuan;Yu, Denghang;Zhou, Zhigang;Wei, Jin;Liu, Lihe;Liu, Jun;Xu, Hongsen;Zhou, Zhigang;Zhou, Zhigang;Liu, Fuguo;Su, Yingbing;Xu, Hongsen

作者机构：

关键词： Pelophylax nigromaculatus; Co-infection; Elizabethkingia miricola; Klebsiella pneumoniae; Whole genome sequence

期刊名称：MICROBIAL PATHOGENESIS （ 影响因子：3.8； 五年影响因子：4.0 ）

ISSN： 0882-4010

年卷期： 2023 年 180 卷

页码：

收录情况： SCI

摘要： Pelophylax nigromaculatus is a common commercial specie of frogs that generally cultured throughout China. With the application of high-density culture, P. nigromaculatus can be co-infected by two or more pathogens, which thereby induce synergistic influence on the virulence of the infection. In this study, two bacterial strains were simultaneously isolated from diseased frogs by incubating on Luria-Bertani (LB) agar. Isolates were identified as Klebsiella pneumoniae and Elizabethkingia miricola by morphological, physiological and biochemical features, as well as 16S rRNA sequencing and phylogenetic analysis. The whole genome of K. pneumoniae and E. miricola isolates consist single circular chromosome of 5,419,557 bp and 4,215,349 bp, respectively. The genomic sequence analysis further indicated that K. pneumoniae isolate conserved 172 virulent and 349 antibiotic-resistance genes, whereas E. miricola contained 24 virulent and 168 antibiotic resistance genes. In LB broth, both isolates could grow well at 0%-1% NaCl concentration and pH 5-7. Antibiotic susceptibility testing revealed that both K. pneumoniae and E. miricola were resistant to kanamycin, neomycin, ampicillin, piperacillin, carbenicillin, enrofloxacin, norfloxacin and sulfisoxazole. Histopathological studies showed that co-infection caused considerable lesions in the tissues of brain, eye, muscle, spleen, kidney and liver, including cell degeneration, necrosis, hemorrhage and inflammatory cell infiltration. The LD50 of K. pneumoniae and E. miricola isolates were 6.31 x 10(5) CFU/g and 3.98 x 10(5) CFU/g frog weight, respectively. Moreover, experimentally infected frogs exhibited quick and higher mortality under coinfection with K. pneumoniae and E. miricola than those single challenge of each bacterium. To date, no natural co-infection by these two bacteria has been reported from frogs and even amphibians. The results will not only shed light on the feature and pathogenesis of K. pneumoniae and E. miricola, but also highlight that co-infection of these two pathogen is a potential threat to black-spotted frog farming.

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