不同地区致奶牛乳房炎金葡菌毒力因子的分布
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摘要
金黄色葡萄球菌(Staphylococcus aureus,SA)是引起奶牛乳房炎的重要病原,该致病菌主要通过毒力因子刺激乳腺组织引起炎症反应。为研究金黄色葡萄球菌重要毒力因子分布的地域性差异,本试验从上海以及云南的3个奶牛场采集奶样和环境污水,利用PCR技术对70株分离菌株和2株标准菌株进行检测。结果显示,在上海地区分离的18株金黄色葡萄球菌中,有7种毒力基因nuc(94.4%)、FnBPA(77.8%)、Hla(88.9%)、Hlb(94.4%)、ClfA(88.9%)、spa(61.1%)和coa(94.4%)的检出率超过50%;在云南分离到的19株金黄色葡萄球菌中,有8种毒力基因nuc(84.2%)、FnBPA(78.9%)、Hla(100%)、Hlb(78.9%)、ClfA(52.6%)、spa(78.9%)、set1 (57.9%)和coa(84.2%)的检出率超过50%,这说明该8种毒力因子可能是致奶牛乳房炎金黄色葡萄球菌中主要的毒力因子。
Staphylococcus aureus(SA) is the main pathogen that could cause dairy cow mastitis.Through its virulence factors,SA can give rise to inflammatory reaction due largely to stimulation of mammary tissues.In order to investigate the regional difference of crucial virulence factors of S.aureus rooted in Shanghai and Yunnan,we collected milk and sewage samples from three dairy farms.Polymerase chain reaction(PCR) was used to detect 2 standard strains and 70 strains isolated from two regions.The detection rates of 7 kinds of virulence factors,including nuc(94.4%),FnBPA(77.8%),Hla(88.9%),Hlb(94.4%),ClfA(88.9%),spa(61.1%) and coa(94.4%),are higher than 50% in18 S.aureus strains isolated from Shanghai.By contrast,the detection rates of 8 kinds of virulence factors,including nuc(84.2%),FnBPA(78.9%),Hla(100%),Hlb(78.9%),ClfA(52.6%),spa(78.9%),set1(57.9%) and coa(84.2%),are higher than 50% in 19 S.aureus strains isolated from Yunnan.The results demonstrate that these 8 virulence factors are probably the dominating virulence factors in dairy cow mastitis.
Staphylococcus aureus(SA) is the main pathogen that could cause dairy cow mastitis.Through its virulence factors,SA can give rise to inflammatory reaction due largely to stimulation of mammary tissues.In order to investigate the regional difference of crucial virulence factors of S.aureus rooted in Shanghai and Yunnan,we collected milk and sewage samples from three dairy farms.Polymerase chain reaction(PCR) was used to detect 2 standard strains and 70 strains isolated from two regions.The detection rates of 7 kinds of virulence factors,including nuc(94.4%),FnBPA(77.8%),Hla(88.9%),Hlb(94.4%),ClfA(88.9%),spa(61.1%) and coa(94.4%),are higher than 50% in18 S.aureus strains isolated from Shanghai.By contrast,the detection rates of 8 kinds of virulence factors,including nuc(84.2%),FnBPA(78.9%),Hla(100%),Hlb(78.9%),ClfA(52.6%),spa(78.9%),set1(57.9%) and coa(84.2%),are higher than 50% in 19 S.aureus strains isolated from Yunnan.The results demonstrate that these 8 virulence factors are probably the dominating virulence factors in dairy cow mastitis.
引文
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[18] Deb R,Kumar A,Chakraborty S,et al.Trends in diagnosis and control of bovine mastitis:a review[J].Pakistan Journal of Biological sciences,2013,16(23):1653-1661.
[19] Hirvonen J J.The use of molecular methods for the detection and identification of methicillin-resistant Staphylococcus aureus[J].Biomarkers in Medicine,2014,8(9):1115-1125.
[2] Magro G,Biffani S,Minozzi G,et al.Virulence genes of S.aureus from dairy cow mastitis and contagiousness risk[J].Toxins,2017,9(6):195-206.
[3] Pyorala S.Indicators of inflammation in the diagnosis of mastitis[J].Veterinary Research,2003,34(5):565-578.
[4] Detilleux J.A strategy to estimate the rate of recruitment of inflammatory cells during bovine intramammary infection under field management[J].BMC Veterinary Research,2017,13(1):167-184.
[5] Cremonesi P,Pozzi F,Raschetti M,et al.Genomic characteristics of Staphylococcus aureus strains associated with high within-herd prevalence of intramammary infections in dairy cows[J].Journal of Dairy Science,2015,98(10):6828-6838.
[6] Ben Said M,Abbassi M S,Bianchini V,et al.Genetic characterization and antimicrobial resistance of Staphylococcus aureus isolated from bovine milk in Tunisia[J].Letters in Applied Microbiology,2016,63(6):473-481.
[7] Salasia S I,Khusnan Z,Lammler C,et al.Comparative studies on pheno- and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany[J].Journal of Veterinary Science,2004,5(2):103-109.
[8] Sinha B,Francois P,Que YA,et al.Heterologously expressed Staphylococcus aureus fibronectin-binding proteins are sufficient for invasion of host cells[J].Infection and Immunity,2000,68(12):6871-6878.
[9] Wang D,Zhang L,Zhou X,et al.Antimicrobial susceptibility,virulence genes,and randomly amplified polymorphic DNA analysis of Staphylococcus aureus recovered from bovine mastitis in Ningxia,China[J].Journal of Dairy Science,2016,99(12):9560-9569.
[10] Foster T J,Geoghegan J A,Ganesh V K,et al.Adhesion,invasion and evasion:the many functions of the surface proteins of Staphylococcus aureus [J].Nature Reviews,2014,12(1):49-62.
[11] Geoghegan J A,Foster T J.Cell wall-anchored surface proteins of Staphylococcus aureus:Many proteins,multiple functions[J].Current Topics in Microbiology and Immunology,2017,409:95-120.
[12] Kim D G,Ahn S H,Kim L H,et al.Application of the rpoS gene for species-specific detection of Vibrio vulnificus by real-time PCR[J].Journal of Microbiology and Biotechnology,2008,18(11):1841-1847.
[13] Park J Y,Fox L K,Seo K S,et al.Comparison of phenotypic and genotypic methods for the species identification of coagulase-negative staphylococcal isolates from bovine intramammary infections[J].Veterinary Microbiology,2011,147(1-2):142-148.
[14] Geha D J,Uhl J R,Gustaferro C A,et al.Multiplex PCR for identification of methicillin-resistant staphylococci in the clinical laboratory[J].Journal of Clinical Microbiology,1994,32(7):1768-1772.
[15] Brakstad O G,Aasbakk K,Maeland J A.Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene[J].Journal of Clinical Microbiology,1992,30(7):1654-1660.
[16] Lina G,Piemont Y,Godail-Gamot F,et al.Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia[J].Clinical Infectious Diseases,1999,29(5):1128-1132.
[17] Cremonesi P,Luzzana M,Brasca M,et al.Development of a multiplex PCR assay for the identification of Staphylococcus aureus enterotoxigenic strains isolated from milk and dairy products[J].Molecular and Cellular Probes,2005,19(5):299-305.
[18] Deb R,Kumar A,Chakraborty S,et al.Trends in diagnosis and control of bovine mastitis:a review[J].Pakistan Journal of Biological sciences,2013,16(23):1653-1661.
[19] Hirvonen J J.The use of molecular methods for the detection and identification of methicillin-resistant Staphylococcus aureus[J].Biomarkers in Medicine,2014,8(9):1115-1125.