中国荷斯坦牛乳房炎病原菌的分离鉴定与分析
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摘要
为了解中国荷斯坦牛乳房炎的流行规律,于2014—2016年采集江苏省某奶牛场荷斯坦牛1 430份乳房炎奶样(有年龄记录的奶样1 046份),采用传统微生物鉴定方法和PCR技术对奶样进行细菌分离鉴定。结果显示,采样奶牛场患乳房炎母牛主要表现为单一感染,主要病原菌为凝固酶阴性葡萄球菌、大肠杆菌和金黄色葡萄球菌,从青年母牛中分离到的病原菌最多,2岁奶牛乳房炎病原菌共分离出428份(40. 9%),其中,凝固酶阴性葡萄球菌298份,占从2岁患乳房炎奶牛中分离出主要致病菌总数的69. 6%; 3岁奶牛乳房炎病原菌分离出548份(52. 4%),其中,237份为凝固酶阴性葡萄球菌、273份为大肠杆菌,分别占从3岁患乳房炎奶牛中分离出主要致病菌总数的43. 2%、49. 8%。病原菌主要分布在2014年(47. 9%)和2015年(49. 7%)。凝固酶阴性葡萄球菌主要分布在夏季(22. 8%)和冬季(13. 2%),大肠杆菌主要分布在秋季207份(14. 5%)。可见,采样奶牛场乳房炎主要发生在青年母牛,乳房炎在夏季、秋季和冬季多发。
In order to explore the epidemic law of Chinese Holstein cow mastitis,1 430 mastitis milk samples were collected from a Holstein dairy farm in Jiangsu province from 2014 to 2016,and traditional microbial identification methods and PCR techniques were used for isolation and identification of bacterial in milk samples. The results showed that most mastitis cows were singly infected by main pathogen,such as coagulase-negative Staphylococci,Escherichia coli and Staphylococcus aureus. The most pathogenic bacteria was isolated from young cows. A total of 428(40. 9%) pathogens causing mastitis were isolated from 2-year-old cow,of which 298(69. 6%) were coagulase-negative Staphylococci,and 548(52. 4%)pathogens were isolated from 3-year-old cow,237(43. 2%) coagulase-negative Staphylococci and 273(49. 8%) Escherichia coli were isolated. The pathogenic bacteria of mastitis were mainly distributed in2014(47. 9%) and 2015(49. 7%). Coagulase-negative Staphylococci were mainly distributed in summer(22. 8%) and winter(13. 2%),and Escherichia coli was mainly distributed in autumn(207,14. 5%). It can be seen that mastitis of sampled dairy cows mainly occurs in young ages,and mastitis occurs frequently in summer,autumn and winter.
In order to explore the epidemic law of Chinese Holstein cow mastitis,1 430 mastitis milk samples were collected from a Holstein dairy farm in Jiangsu province from 2014 to 2016,and traditional microbial identification methods and PCR techniques were used for isolation and identification of bacterial in milk samples. The results showed that most mastitis cows were singly infected by main pathogen,such as coagulase-negative Staphylococci,Escherichia coli and Staphylococcus aureus. The most pathogenic bacteria was isolated from young cows. A total of 428(40. 9%) pathogens causing mastitis were isolated from 2-year-old cow,of which 298(69. 6%) were coagulase-negative Staphylococci,and 548(52. 4%)pathogens were isolated from 3-year-old cow,237(43. 2%) coagulase-negative Staphylococci and 273(49. 8%) Escherichia coli were isolated. The pathogenic bacteria of mastitis were mainly distributed in2014(47. 9%) and 2015(49. 7%). Coagulase-negative Staphylococci were mainly distributed in summer(22. 8%) and winter(13. 2%),and Escherichia coli was mainly distributed in autumn(207,14. 5%). It can be seen that mastitis of sampled dairy cows mainly occurs in young ages,and mastitis occurs frequently in summer,autumn and winter.
引文
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[14] Xu J,Tan X,Zhang X,et al. The diversities of staphylococcal species,virulence and antibiotic resistance genes in the subclinical mastitis milk from a single Chinese cow herd[J]. Microbial Pathogenesis,2015,88:29-38.
[15]林伟东,宰栩生,于恩琪,等.江苏地区奶牛乳房炎凝固酶阴性葡萄球菌耐药基因分析[J].中国兽医学报,2015,35(7):1130-1135.
[16] Pyorala,Taponen S. Coagulase-negative staphylococci—Emerging mastitis pathogens[J]. Veterinary Microbiology,2009,134(1/2):3-8.
[17] Tenhagen B A,Koster G,Wallmann J,et al. Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg,Germany[J]. Journal of Dairy Science,2006,89(7):2542-2551.
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[19] Koivula M,Pitkala A,Pyorala S,et al. Distribution of bacteria and seasonal and regional effects in a new database for mastitis pathogens in Finland[J]. Acta Agriculturae Scand Section A,2007,57(2):89-96.
[2]高鹏飞,程斌,张善亭,等.复合乳酸菌制剂在防治奶牛乳房炎方面的应用研究[J].中国畜牧杂志,2014,50(12):41-47.
[3] Ruegg P L. Investigation of mastitis problems on farms[J]. Veterinary Clinics of North America Food Animal Practice,2003,19(1):47-73.
[4]聂培.奶牛乳房炎病原菌分离鉴定及其多重PCR检测方法的建立[D].南宁:广西大学,2013.
[5] Shome B R,Mitra S D,Bhuvana M,et al. Multiplex PCR assay for species identification of bovine mastitis pathogens[J]. Journal of Applied Microbiology,2011,111(6):1349-1356.
[6]田彦.基于高通量测序的中国荷斯坦奶牛瘤胃宏转录组研究[D].北京:中国科学院北京基因组研究所,2015.
[7]徐玉薇,罗小青,魏学良,等.不同地区3个奶牛场临床型乳房炎细菌病原种类分析[J].中国畜牧兽医,2016,43(2):548-552.
[8]罗金印,李宏胜,李新圃,等.兰州地区奶牛乳房炎流行病学及病原菌调查[J].黑龙江畜牧兽医,2015(21):164-166.
[9]韩淑芳,刘一飞,王国艳,等.某奶牛场牛乳源金黄色葡萄球菌的分离鉴定与耐药性调查[J].山西农业科学,2016,44(11):1696-1698.
[10]布登付.奶牛乳房炎的发生与防治[J].河南农业科学,2000(7):37.
[11] Walcher M,Skvoretz R,Montgomery-fullerton M,et al.Description of an unusual neisseria meningitidis isolate containing and expressing neisseria gonorrhoeae-specific16S rRNA gene sequences[J]. Journal of Clinical Microbiology,2013,51(10):3199-3206.
[12]苏少虎,李冉.新疆地区奶牛乳房炎病原菌的流行病学调查[J].中国动物保健,2014,16(12):20-23.
[13]孙垚,贾坤,平晓坤,等.广东省部分地区奶牛乳房炎凝固酶阴性葡萄球菌的毒力基因和耐药性分析[J].中国兽医学报,2017,37(8):1495-1500.
[14] Xu J,Tan X,Zhang X,et al. The diversities of staphylococcal species,virulence and antibiotic resistance genes in the subclinical mastitis milk from a single Chinese cow herd[J]. Microbial Pathogenesis,2015,88:29-38.
[15]林伟东,宰栩生,于恩琪,等.江苏地区奶牛乳房炎凝固酶阴性葡萄球菌耐药基因分析[J].中国兽医学报,2015,35(7):1130-1135.
[16] Pyorala,Taponen S. Coagulase-negative staphylococci—Emerging mastitis pathogens[J]. Veterinary Microbiology,2009,134(1/2):3-8.
[17] Tenhagen B A,Koster G,Wallmann J,et al. Prevalence of mastitis pathogens and their resistance against antimicrobial agents in dairy cows in Brandenburg,Germany[J]. Journal of Dairy Science,2006,89(7):2542-2551.
[18] Ostera s O,Solverod L,Reksen O. Milk culture results in a large Norwegian survey—Effects of season,parity,days in milk,resistance,and clustering[J]. Journal of Dairy Science,2006,89(3):1010-1023.
[19] Koivula M,Pitkala A,Pyorala S,et al. Distribution of bacteria and seasonal and regional effects in a new database for mastitis pathogens in Finland[J]. Acta Agriculturae Scand Section A,2007,57(2):89-96.