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一株源自孔雀的新型支原体的鉴定及其致病性和基因组分析
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国家重点研发计划(2016YFD0500800);山东省重点研发计划(2018GNC110020);山东省自然科学基金(ZR2017MC038)


Identification, pathogenicity and genome analysis of a new mycoplasma from peacock
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    摘要:

    【背景】Mycoplasma gallinaceum (MGC)是禽源支原体的一种,仅在南非、英国等地发生,我国鲜有报道。【目的】从患呼吸道病的孔雀气管中分离到一株支原体,命名为Peacock20181011,确定其分类、致病性和基因组特征。【方法】利用微生物学常规方法,结合分子生物学检测和基因组序列,对其进行鉴定;通过对Special pathogenic free (SPF)鸡、鸡胚的致病性研究和最小抑菌浓度(minimal inhibitory concentration,MIC)测定,确定其致病性和药敏性。【结果】通过对分离菌的培养、纯化、形态学和染色观察,结合生化试验和16S rRNA基因测序证实,该菌为一株新的MGC,与模式菌株NCTC10183的16S rRNA基因相似性高达99.86%,系统发育树显示该菌与MGC属同一分支;人工感染实验表明该菌对SPF鸡无致病性,但可造成SPF鸡胚发育迟缓,爪部蜷缩;MIC结果显示该菌对单硫酸卡那霉素和氟苯尼考等敏感。基因组序列表明,该菌基因组长度为1 183 913 bp,(G+C)mol%含量为28.7%,含有898个Coding sequences (CDS),拥有4个拷贝的16S rRNA基因和6个质粒,预测有20个毒力因子基因和2个耐药基因。【结论】明确了MGC在我国的存在,丰富了中国禽源支原体的种类,为支原体病的防控提供了依据。

    Abstract:

    [Background] Mycoplasma gallinaceum is a kind of mycoplasma in poultry, which occurs only in South Africa and other places, but rarely reported in China. [Objective] A mycoplasma strain was isolated from the trachea of peacock with respiratory tract disease and named Peacock20181011, to determine its taxonomic status, pathogenicity and molecular characteristics. [Methods] The newly isolated pathogenic bacterium was identified by routine microbial methods, combined with molecular biological methods and full-length genome sequences. The pathogenicity and drug sensitivity of the newly isolated pathogens were determined by pathogenicity in special pathogenic free chickens, embryos and minimal inhibitory concentration test. [Results] Through the culture, purification, morphology and staining observation of isolates, combined with biochemical tests and 16S rRNA gene sequencing, it was confirmed that Peacock20181011 belonged to Mycoplasma gallinaceum, and the 16S rRNA gene similarity to that of standard strain NCTC10183 (LR214950) was up to 99.86%, and the phylogenetic tree showed that Peacock20181011 was in the same branch of Mycoplasma gallinaceum; Artificial infection experiment showed that the fungus was not pathogenic to special pathogenic free chicken, but could cause special pathogenic free chicken embryos to develop late and the claw was curled up; minimal inhibitory concentration test showed that the bacterium was sensitive to kanamycin and florfenicol. The analysis of genome sequence showed that the bacterial length was 1 183 913 bp and the content of (G+C)mol% was 28.7%, which contained 898 coding sequences with four copies of 16S rRNA gene. [Conclusion] The existence of Mycoplasma gallinaceum in China is defined, the species of Mycoplasma in China are enriched, and the basis for prevention and control of the disease is provided.

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刘星丽,徐怀英,仉伟,黄迪海,秦春芝,郭卉,秦卓明. 一株源自孔雀的新型支原体的鉴定及其致病性和基因组分析[J]. 微生物学通报, 2020, 47(6): 1828-1836

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  • 在线发布日期: 2020-06-01
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