高通量测序法检测奶牛乳房炎关联微生物群落结构及多样性

doi:10.3969/j.issn.1004-1524.2019.09.06

浙江农业学报 ›› 2019, Vol. 31 ›› Issue (9): 1437-1445.DOI: 10.3969/j.issn.1004-1524.2019.09.06

高通量测序法检测奶牛乳房炎关联微生物群落结构及多样性

曾学琴, 柳陈坚, 杨雪, 李晓然^*

昆明理工大学生命科学与技术学院,云南昆明 650500

收稿日期:2019-04-05 出版日期:2019-09-25 发布日期:2019-10-11
通讯作者: *李晓然,E-mail: starkeyran@163.com
作者简介:曾学琴(1993—),女,云南文山人,硕士研究生,主要从事微生物学相关研究。E-mail: 1711169659@qq.com

Microbial community structure and diversity of mastitis cows by 16S rRNA high-throughput sequencing

ZENG Xueqin, LIU Chenjian, YANG Xue, LI Xiaoran^*

Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China

Received:2019-04-05 Online:2019-09-25 Published:2019-10-11

摘要/Abstract

摘要： 为对引起奶牛乳房炎的病原菌进行溯源,对乳房炎奶牛和正常奶牛的乳头擦拭子和牛奶中细菌的16S rRNA基因的V4-V5高变区进行扩增和高通量测序,使用生物统计学方法分析微生物多样性。结果显示,乳房炎奶牛乳头和牛奶中微生物组成呈现明显差异:在门的水平上,乳头丰度最高的是厚壁菌门(Firmicutes),牛奶则是变形菌门(Proteobacteria),乳头样品均含有拟杆菌门(Bacteroidetes)细菌,而牛奶中几乎没有,梭杆菌门(Fusobacteria)细菌仅存在于乳头中;在属的水平上,梭杆菌属(Fusobacterium)仅在乳头上检测到,牛奶中丰度最高的为假单胞菌属(Pseudomonas)细菌,其仅存在于2个乳头样品中,不动杆菌属(Acinetobacter)细菌在两类样品中都有,但乳头的含量高于牛奶。奶牛患乳房炎后乳头和牛奶的微生物群落与正常奶牛也有差异:梭杆菌门(Fusobacteria)仅在乳房炎奶牛乳头中检测到,正常奶牛没有,乳房炎奶牛乳头金黄色酿脓葡萄球菌属(Staphylococcus)丰度高于正常奶牛,肠球菌属(Enterococcus)仅在乳房炎奶牛牛奶中检测到,正常奶牛没有。奶牛乳头与牛奶中的微生物群落差异极大,奶牛患乳房炎后乳房外和乳房内微生物组成都会发生变化,外源病原菌和内源病原菌都可能引起乳房炎。

关键词: 奶牛乳房炎, 乳头, 牛奶, 微生物多样性

Abstract: To clarify the pathogens that cause mastitis in cows, the V4-V5 hypervariable region of bacterial 16S rRNA gene in nipple wipes and milk of mastitis cows and normal cows were amplified and high throughput sequenced. The succession of microbial diversity was analyzed by biostatistics. The results showed that there was a very obvious difference in the composition of microbial between the nipple and the milk of the mastitis cow. At the phylum level, the highest abundance was Firmicutes in the nipples, while Proteobacteria was the highest in the milk. All nipples contain Bacteroidetes, while there is almost no in the milk, Fusobacteria only exists in nipples. The microbial community of the nipple and milk after mastitis in cows was different from normal cows. Fusobacteria was only detected on the nipples of mastitis cows, and normal cows were not. The abundance of Staphylococcus on the nipples of mastitis cows was higher than that of normal cows. Enterococcus was detected in mastitis cow milk, but not in normal cows. Therefore, the nipple of the cow was very different from the microbial community in the milk. Microbial composition changes both outside and inside the breast after bovine mastitis. Exogenous pathogens and endogenous sources pathogens can cause mastitis.

Key words: bovine mastitis, nipple, milk, microbial diversity

中图分类号:

S852.61

曾学琴, 柳陈坚, 杨雪, 李晓然. 高通量测序法检测奶牛乳房炎关联微生物群落结构及多样性[J]. 浙江农业学报, 2019, 31(9): 1437-1445.

ZENG Xueqin, LIU Chenjian, YANG Xue, LI Xiaoran. Microbial community structure and diversity of mastitis cows by 16S rRNA high-throughput sequencing[J]. , 2019, 31(9): 1437-1445.

参考文献

[1] GOMES F, SAAVEDRA M J, HENRIQUES M.Bovine mastitis disease/pathogenicity: evidence of the potential role of microbial biofilms[J]. Pathogens and Disease, 2016: ftw006.
[2] HALASA T, HUIJPS K, ØSTERÅS O, et al. Economic effects of bovine mastitis and mastitis management: a review[J]. Veterinary Quarterly, 2007, 29(1): 18-31.
[3] CHA E, BAR D, HERTL J A, et al.The cost and management of different types of clinical mastitis in dairy cows estimated by dynamic programming[J]. Journal of Dairy Science, 2011, 94(9): 4476-4487.
[4] COULONA J, GASQUIB P, BARNOUIN J, et al.Effect of mastitis and related-germ on milk yield and composition during naturally-occurring udder infections in dairy cows[J]. Animal Research, 2002, 51(5): 383-393.
[5] HAMBALI I U, BHUTTO K R, JESSE F F A, et al. Clinical responses in cows vaccinated with a developed prototype killed Staphylococcus aureus mastitis vaccine[J]. Microbial Pathogenesis, 2018, 124: 101-105.
[6] 庆麦玉. 奶牛的乳房保健[J]. 中国奶牛, 2000(2): 51-53.
QING M Y.Breast health care of dairy cows[J]. China Dairy Cattle, 2000(2):51-53.(in Chinese)
[7] 胡松华, 蒋次升. 奶牛隐性乳房炎对牛奶脂肪、蛋白质和乳糖含量的影响[J]. 中国奶牛, 1990(3): 42-43.
HU S H, JIANG C S.Effect of recessive mastitis on milk fat, protein and lactose content in dairy cows[J]. China Dairy Cattle, 1990(3): 42-43. (in Chinese)
[8] 李宏胜, 郁杰, 李新圃. 奶牛乳房炎病原菌区系分布及发病规律研究进展[J]. 动物医学进展, 2004, 26(6): 146-149.
LI H S, YU J, LI X P.Research progress on flora distribution and pathogenesis of mastitis in dairy cows[J]. Progress in Veterinary Medicine, 2004, 26(6):146-149.(in Chinese with English abstract)
[9] 袁永隆, 张礼华, 刘纯传, 等. 我国奶牛乳房炎常见病原菌的区系调查[J]. 中国农业科学, 1992, 25(4): 70-76.
YUAN Y L, ZHANG L H, LIU C C, et al.A survey of the pathogens of bovine mastitis in China[J]. Scientia Agricultura Sinica, 1992, 25(4): 70-76.(in Chinese with English abstract)
[10] 周庆新, 李锦春, 雷彬. 奶牛隐性乳房炎的病原分离及药敏试验报告[J]. 今日畜牧兽医, 2007 (4): 4-6.
ZHOU Q X, LI J C, LEI B.Pathogen isolation and drug sensitivity test of recessive mastitis in dairy cows[J]. Today Animay Husbandry and Veterinary Medicine, 2007(4): 4-6.(in Chinese)
[11] 迪沃斯,赵德明. 奶牛疾病学 [M]. 北京: 中国农业大学出版社, 2009.
[12] TAPONEN S, SALMIKIVI L, SIMOJOKI H, et al.Real-time polymerase chain reaction-based identification of bacteria in milk samples from bovine clinical mastitis with no growth in conventional culturing[J]. Journal of Dairy Science, 2009, 92(6):2610-2617.
[13] HAMBALI I, AHMED M, ADAMU N, et al.PCR-based detection of Schistosoma bovis in cattle in Maiduguri Metropolis and Jere Local Government Areas in Borno State, Nigeria[J]. Journal of Advanced Veterinary and Animal Research, 2016, 3(2): 92-98.
[14] SCHMIDT T M, DELONG E F, PACE N R.Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing[J]. Journal of Bacteriology, 1991, 173(14): 4371-4378.
[15] SCHLOSS P D, WESTCOTT S L, RYABIN T, et al.Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities[J]. Applied and Environmental Microbiology, 2009, 75(23): 7537-7541.
[16] PRUESSE E, QUAST C, KNITTEL K, et al.SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB[J]. Nucleic Acids Research, 2007, 35(21): 7188-7196.
[17] THOMPSON J.The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 1997, 25(24): 4876-4882.
[18] TAMURA K, PETERSON D, PETERSON N, et al.MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Molecular Biology and Evolution, 2011, 28(10): 2731-2739.
[19] SAITOU N, NEI M.The neighbor-joining method: a new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 1987, 4(4):406-425.
[20] CATOZZI C, SANCHEZ BONASTRE A, FRANCINO O, et al.The microbiota of water buffalo milk during mastitis[J]. PLoS One, 2017, 12(9): e0184710.
[21] OKUDA T, KOKUBU E, KAWANA T, et al.Synergy in biofilm formation between Fusobacterium nucleatum and Prevotella species[J]. Anaerobe, 2012, 18(1): 110-116.
[22] SAITO Y, FUJII R, NAKAGAWA K I, et al.Stimulation of Fusobacterium nucleatum biofilm formation by Porphyromonas gingivalis[J]. Oral Microbiology and Immunology, 2008, 23(1):1-6.
[23] SWIDSINSKI A, DORFFEL Y, LOENING-BAUCKE V, et al.Acute appendicitis is characterised by local invasion with Fusobacterium nucleatum/necrophorum[J]. Gut, 2011, 60(1): 34-40.
[24] PELEG A Y, SEIFERT H, PATERSON D L.Acinetobacter baumannii: emergence of a successful pathogen[J]. Clinical Microbiology Reviews, 2008, 21(3): 538-582.
[25] TOSAKI K, KOJIMA H, AKAMA S, et al.Bovine esophageal and glossal ulceration associated with Pseudomonas aeruginosa and Fusobacterium spp. in a 10-month-old Holstein heifer[J]. The Journal of Veterinary Medical Science, 2018, 80(7): 1174-1178.
[26] 江涛, 陈虹, 罗永艾. 医院内嗜麦芽寡养单胞菌肺部感染的临床调查[J]. 中华医院感染学杂志, 2003, 13(2): 177-179.
JIANG T, CHEN H, LUO Y A.Hospital lung infection by Stenotrophomonas maltophilia: a clinical investigation[J]. Chinese Journal of Nosocomiology, 2003, 13(2): 177-179.(in Chinese)
[27] TALBOT B G, LACASSE P.Progress in the development of mastitis vaccines[J]. Livestock Production Science, 2005, 98(1/2): 101-113.
[28] BATTY A, WREN M W D, GAL M. Fusobacterium necrophorum as the cause of recurrent sore throat: comparison of isolates from persistent sore throat syndrome and Lemierre's disease[J]. Journal of Infection, 2005, 51(4): 299-306.

编辑推荐

Metrics

阅读次数

全文

1877

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	0	0	0	1877

来源	本网站	其他网站

次数	1225	652
比例	65%	35%

摘要

749

最新录用	在线预览	正式出版

0	0	749

来源	本网站	其他网站

次数	747	2
比例	100%	0%

高通量测序法检测奶牛乳房炎关联微生物群落结构及多样性

Microbial community structure and diversity of mastitis cows by 16S rRNA high-throughput sequencing

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

编辑推荐

Metrics

本文评价

[1]	徐民民, 黄莹, 李波, 徐艳, 张帅, 姚岭芸, 王政. 生物炭对小麦根际和根内微生物群落结构的影响[J]. 浙江农业学报, 2021, 33(3): 516-525.
[2]	白东东, 李新圃, 杨峰, 罗金印, 王旭荣, 李宏胜. 地肤通乳散治疗奶牛乳房炎临床疗效及作用靶点预测[J]. 浙江农业学报, 2019, 31(5): 730-736.
[3]	刘祥, 孔智翔, 殷书平, 殴莎莎, 刘成艳, 同韩虎, 吴三桥, 陈琛, 陈春琳. 奶牛乳房炎重要候选疫苗蛋白的抗原表位分析及三联重组表位疫苗的氨基酸序列设计[J]. 浙江农业学报, 2016, 28(9): 1476-1484.
[4]	熊毅;郭彦萃;张志. 原始红松林与人工林土壤微生物多样性分析[J]. , 2014, 26(1): 0-164.
[5]	王阳*;王奇赞 . 种植年限对大棚蔬菜地土壤微生物群落结构多样性的影响[J]. , 2013, 25(3): 0-576.