[1]潘子豪,龚倩梅,李科茫,等.奶牛乳液中伊丽莎白菌按蚊亚种高度同源株的分离与鉴定[J].南京农业大学学报,2017,40(6):1088-1093.[doi:10.7685/jnau.201709019]
 PAN Zihao,GONG Qianmei,LI Kemang,et al.Isolation and identification of a bacterium from milk of a dairy cow with mastitis sharing high homology to Elizabethkingia anophelis[J].Journal of Nanjing Agricultural University,2017,40(6):1088-1093.[doi:10.7685/jnau.201709019]
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奶牛乳液中伊丽莎白菌按蚊亚种高度同源株的分离与鉴定()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
40卷
期数:
2017年6期
页码:
1088-1093
栏目:
出版日期:
2017-11-10

文章信息/Info

Title:
Isolation and identification of a bacterium from milk of a dairy cow with mastitis sharing high homology to Elizabethkingia anophelis
作者:
潘子豪 龚倩梅 李科茫 马慧 吴宗福 姚火春
南京农业大学动物医学院, 江苏 南京 210095
Author(s):
PAN Zihao GONG Qianmei LI Kemang MA Hui WU Zongfu YAO Huochun
College of Veterinary Meidicne, Nanjing Agricultural University, Nanjing 210095, China
关键词:
伊丽莎白菌伊丽莎白菌类按蚊亚种奶牛乳房炎
Keywords:
ElizabethkingiaElizabethkingia anophelisdairy cowmastitis
分类号:
Q939.96
DOI:
10.7685/jnau.201709019
摘要:
[目的] 本文旨在从患乳房炎的奶牛乳液中分离鉴定病原菌。[方法] 收集患乳房炎奶牛的乳液病料后分别接种绵羊血液平板和麦康凯鉴别培养基,置于37℃培养箱过夜。观察获得的单菌落形态并接种含有血清的液体培养基富集培养,然后煮沸法提取细菌纯培养物的基因组,以该基因组为模板通过PCR扩增未知细菌的核糖体RNA序列并对产物进行测序分析。以不同培养时间测定菌液D600并绘制细菌的生长曲线。用倍比稀释法在96孔板上测定细菌对不同药物的最小抑菌浓度。[结果] 从病料中分离到一株革兰阴性菌,命名为JS20170427COW。该菌无运动性,在绵羊血平板上菌落圆形、光滑、淡黄色、边界清晰。生化试验和16S rRNA序列分析结果显示,其与伊丽莎白按蚊亚种人源株KT768343.1同源性达95%,称之为伊丽莎白按蚊亚种高度同源株。药敏试验显示,该菌对杆菌肽、溶菌素、硫酸黏菌素、乳酸菌素以及多肽类药物等耐药。[结论] 从病牛乳液中分离到一株与伊丽莎白菌按蚊亚种同源性高达95%的细菌,其具有公共卫生的潜在意义。
Abstract:
[Objectives] To identify the pathogens for mastitis in dairy cows, the bacteria were isolated from milk of dairy cows with mastitis.[Methods] Samples were plated onto blood agar plates and macconkey nutrition agar plates using sterile loops then were incubated for 24 h at 37℃. The characters of the colony form were observed and fostered in liquid culture with serum. The isolates were subjected to 16S rRNA PCR to ensure isolates species through sequence alignment in NCBI database. According the results of cell counting D600 value, the bacterium growth curve was made. Minimal inhibitory concentrations (MIC) for the inoculum assay was defined as the lowest concentration that caused a 50% inhibition of bacterial growth in 96-well plate.[Results] A Gram-negative bacterium (named JS20170427COW)was isolated from milk of a dairy cow with mastitis. Biochemical characteristic analysis showed the bacterium is non-motile. Colonies on sheep-blood agar are smooth, yellowish, circular and shiny with entire edges. Sequence homology analysis indicated that 16S rRNA gene of the bacterium shares 95% homology with Elizabethkingia anopheles strain KT768343. 1 isolated from human. Thus, strain JS20170427COW was designated as Elizabethkingia anopheles highly homological strain. Drug resistance test indicated that JS20170427COW is resistant to the bacterin, lysin, sulfuric acid, lactobacillin and multi-peptides antibiotics.[Conclusions] The bacterium, JS20170427COW, sharing high homology with Elizabethkingia anopheles was identified and isolated from milk of a dairy cow with mastitis, which shows the potential significance to public health.

参考文献/References:

[1] Kim K K,Kim M K,Lim J H,et al. Transfer of Chryseobacterium meningosepticum and Chryseobacterium miricola to Elizabethkingia gen. nov. as Elizabethkingia meningoseptica comb. nov. and Elizabethkingia miricola comb. nov[J]. International Journal of Systematic and Evolutionary Microbiology,2005,55:1287-1293.
[2] Kampfer P,Matthews H,Glaeser S P,et al. Elizabethkingia anophelis sp. nov. isolated from the midgut of the mosquito Anopheles gambiae[J]. International Journal of Systematic and Evolutionary Microbiology,2011,61:2670-2675.
[3] Frank T,Gody J C,Nguyen L B,et al. First case of Elizabethkingia anophelis meningitis in the Central African Republic[J]. Lancet,2013,381:1876.
[4] Lau S K,Wu A K,Teng J L,et al. Evidence for Elizabethkingia anophelis transmission from mother to infant,Hong Kong[J]. Emerging Infectious Diseases,2015,21(2):232-241.
[5] Lin J N,Yang C H,Lai C H,et al. Draft genome sequence of Elizabethkingia anophelis strain EM361-97 isolated from the blood of a cancer patient[J]. Genome Announcements,2016,4(5):e01215-16.
[6] Navon L,Clegg W J,Morgan J,et al. Notes from the field:investigation of Elizabethkingia anophelis cluster-illinois,2014-2016[J]. Morbidity and Mortality Weekly Report,2016,65(48):1380-1381.
[7] Nicholson A C,Whitney A M,Emery B D,et al. Complete genome sequences of four strains from the 2015-2016Elizabethkingia anophelis outbreak[J]. Genome Announcements,2016,4(3):e00563-16.
[8] Coyle A L. Elizabethkingia anophelis:exploring the outbreak of disease in the Midwest[J]. Nursing,2017,47(3):61-63.
[9] Lau S K,Chow W N,Foo C H,et al. Elizabethkingia anophelis bacteremia is associated with clinically significant infections and high mortality[J]. Scientific Reports,2016,6:26045.
[10] Doijad S,Ghosh H,Glaeser S,et al. Taxonomic reassessment of the genus Elizabethkingia using whole-genome sequencing:Elizabethkingia endophytica Kampfer et al. 2015 is a later subjective synonym of Elizabethkingia anophelis Kampfer et al. 2011[J]. International Journal of Systematic and Evolutionary Microbiology,2016,66(11):4555-4559.
[11] Perrin A,Larsonneur E,Nicholson A C,et al. Evolutionary dynamics and genomic features of the Elizabethkingia anophelis 2015 to 2016 Wisconsin outbreak strain[J]. Nature Communications,2017,8:15483.
[12] Janda J M,Lopez D L.Mini review:new pathogen profiles:Elizabethkingia anophelis[J]. Diagnostic Microbiology and Infectious Disease,2017,88(2):201-205.
[13] Hu S,Jiang T,Zhang X,et al. Elizabethkingia anophelis isolated from patients with multiple organ dysfunction syndrome and lower respiratory tract Infection:report of two cases and literature review[J]. Frontiers in Microbiology,2017,8:382.
[14] Garay J A R,Hughes G L,Koundal V,et al. Genome sequence of Elizabethkingia anophelis strain EaAs1,isolated from the Asian Malaria Mosquito Anopheles stephensi[J]. Genome Announcements,2016,4(2):e00084-16.
[15] Breurec S,Criscuolo A,Diancourt L,et al. Genomic epidemiology and global diversity of the emerging bacterial pathogen Elizabethkingia anophelis[J]. Scientific Reports,2016,6:30379.
[16] Teo J,Tan S Y,Liu Y,et al. Comparative genomic analysis of malaria mosquito vector-associated novel pathogen Elizabethkingia anophelis[J]. Genome Biology and Evolution,2014,6(5):1158-1165.
[17] Han M S,Kim H,Lee Y,et al. Relative prevalence and antimicrobial susceptibility of clinical isolates of Elizabethkingia species based on 16S rRNA gene sequencing[J]. Journal of Clinical Microbiology,2017,55(1):274-280.

备注/Memo

备注/Memo:
收稿日期:2017-09-11。
基金项目:南京农业大学创新性实践教学项目(2017sy22)
作者简介:潘子豪,讲师,主要从事兽医微生物与免疫学研究,E-mail:panzihao@njau.edu.cn。
通信作者:姚火春,教授,主要从事兽医微生物与免疫学研究,E-mail:yaohch@njau.edu.cn。
更新日期/Last Update: 1900-01-01