奶牛源金黄色葡萄球菌临床分离株的耐药特性、生物膜及其相关基因分布研究
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摘要
【目的】为了解引起奶牛乳房炎的金黄色葡萄球菌(SA)分离株的耐药性及其生物膜特性。【方法】本研究采用纸片扩散法对新疆地区奶牛源临床分离的164株SA的耐药特性进行检测,并对MRSA进行判定;用BHI培养24 h形成生物膜,结晶紫染色后测其吸光度(OD570 nm)值,比较分析MRSA和MSSA菌株间生物膜形成能力;对生物膜形成相关基因的分布进行检测。【结果】164株SA临床分离株中共检出22株MRSA,检出率为13.41%;SA对青霉素、头孢西丁、红霉素、甲氧苄啶、四环素耐药株较多;MRSA对头孢类、氟喹诺酮类和大环内酯类抗菌药物的耐药率明显高于甲氧西林敏感SA(MSSA);24 h时,MRSA和MSSA生物膜生成能力差异显著(P<0.05),MRSA菌株的生物膜生成能力显著高于MSSA菌株;MRSA菌株黏附素clf A、clf B、fnb B、Fib、cna、Fn BP基因携带率在54.55%~100%之间,显著高于MSSA菌株检出率(P<0.05)。【结论】呋喃妥因、替考拉宁、磷霉素可作为优选药用于治疗MRSA和β-内酰胺类耐药株引起的严重感染。MRSA菌株较于MSSA菌株具有较强的生物膜形成能力,这与MRSA的较强抗性具有相关性。
【Objective】The present paper was conducted to know the antimicrobial resistance and biofilm characteristics of Staphylococcus aureus( SA) from cow mastitis.【Method】The drug resistance of 164 clinical S. aureu isolates was determined by using disk diffusion method in the present study. Then methicillin-resistant Staphylococcus aureus( MRSA) strains were identified. MRSA and MSSA strains were cultured on BHI for 24 hours respectively,the optical density( OD570 nm) was determined by using a mieroplate reader after staining with crystal violet,then formation capacity of biofilm between MRSA and MSSA strains were comparatively analyzed. The distribution of genes related to biofilm formation was also examined. 【Result】A total of 22 strains of MRSA have been detected among the 164 S. aureu sisolates,with the detection rate of 13. 41 %; To penicillin,cefoxitin,erythromycin,trimethoprim and tetracycline resistant strains of Staphylococcus aureus,the drug resistance rates of MRSA to cephalosporins,quinolones and macrolides antibiotics were significantly higher than those of the methicillin-sensitive Staphylococcus aureus( MSSA),respectively. At 24 hours,the biofilm formation ability of MRSA and MSSA was significantly different( P < 0. 05),and the biofilm formation ability of MRSA strain was significantly higher than that of MSSA strain.The detection rates of adhesin clf A,clf B,fnb B,Fib,cna and Fn BP ranged from 54. 55 % to 100 %,respectively,while the detection rate of MRSA was higher than that of MSSA( P < 0.05). 【Conclusion】Furantoin,teicoplanin and fosfomycin can be used as preferred drugs for the treatment of severe infections caused by MRSA and β-lactam resistant strains. The biofilm formation ability of MRSA strain was higher than that of MSSA strain,which is related to the strong resistance of MRSA.
【Objective】The present paper was conducted to know the antimicrobial resistance and biofilm characteristics of Staphylococcus aureus( SA) from cow mastitis.【Method】The drug resistance of 164 clinical S. aureu isolates was determined by using disk diffusion method in the present study. Then methicillin-resistant Staphylococcus aureus( MRSA) strains were identified. MRSA and MSSA strains were cultured on BHI for 24 hours respectively,the optical density( OD570 nm) was determined by using a mieroplate reader after staining with crystal violet,then formation capacity of biofilm between MRSA and MSSA strains were comparatively analyzed. The distribution of genes related to biofilm formation was also examined. 【Result】A total of 22 strains of MRSA have been detected among the 164 S. aureu sisolates,with the detection rate of 13. 41 %; To penicillin,cefoxitin,erythromycin,trimethoprim and tetracycline resistant strains of Staphylococcus aureus,the drug resistance rates of MRSA to cephalosporins,quinolones and macrolides antibiotics were significantly higher than those of the methicillin-sensitive Staphylococcus aureus( MSSA),respectively. At 24 hours,the biofilm formation ability of MRSA and MSSA was significantly different( P < 0. 05),and the biofilm formation ability of MRSA strain was significantly higher than that of MSSA strain.The detection rates of adhesin clf A,clf B,fnb B,Fib,cna and Fn BP ranged from 54. 55 % to 100 %,respectively,while the detection rate of MRSA was higher than that of MSSA( P < 0.05). 【Conclusion】Furantoin,teicoplanin and fosfomycin can be used as preferred drugs for the treatment of severe infections caused by MRSA and β-lactam resistant strains. The biofilm formation ability of MRSA strain was higher than that of MSSA strain,which is related to the strong resistance of MRSA.
引文
[1]Li T,Lu H,Wang X,et al.Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015[J].Front Cell Infect Microbiol,2017(7):127.
[2]Mcdougall S,Hussein H,Petrovski K.Antimicrobial resistance in Staphylococcus aureus,Streptococcus uberis and Streptococcus dysgalactiae from dairy cows with mastitis[J].New Zealand Veterinary Journal,2014,62(2):68-76.
[3]杨峰,王旭荣,李新圃,等.牛源金黄色葡萄球菌耐药性与相关耐药基因和菌株毒力基因的相关性研究[J].中国兽医科学,2016(2):247-252.
[4]金耀忠,万世平,姜法铭,等.奶牛乳房炎流行情况调查及主要病原分离与药敏试验[J].畜牧与兽医,2011,43(5):64-67.
[5]Serray B,Oufrid S,Hannaoui I,et al.Genes encoding adhesion factors and biofilm formation in methicillin-resistant Staphylococcus aureus in Morocco[J].J Infect Dev Ctries,2016,10(8):863.
[6]Gogoi-Tiwari J,Williams V,Waryah C B,et al.Mammary Gland Pathology Subsequent to Acute Infection with Strong versus Weak Biofilm Forming Staphylococcus aureus Bovine Mastitis Isolates:A Pilot Study Using Non-Invasive Mouse Mastitis Model[J].Plos One,2017,12(1):17-68.
[7]Cha J O,Yoo J I,Yoo J S,et al.Investigation of Biofilm Formation and its Association with the Molecular and Clinical Characteristics of Methicillin-resistant Staphylococcus aureus[J].Osong Public Health Res Perspect,2014,5(2):116.
[8]Nourbakhsh F,Namvar A E.Detection of genes involved in biofilm formation in Staphylococcus aureus isolates[J].GMS Hyg Infect Control,2016,11.
[9]Yang Y,Zhang X,Huang W,et al.SCCmec-associated psm-mec,mRNA promotes Staphylococcus epidermidis,biofilm formation[J].Antonie Van Leeuwenhoek,2016,109(10):1403-1415.
[10]Pereyra E A,Picech F,Renna M S,et al.Detection of Staphylococcus aureus adhesion and biofilm-producing genes and their expression during internalization in bovine mammary epithelial cells[J].Vet Microbiol,2016,183:69-77.
[11]Coelho S M,Reinoso E,Pereira I A,et al.Virulence factors and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Rio de Janeiro[J].Pesquisa Veterinária Brasileira,2009,29(5):369-374.
[12]Li J,Feβler A T,Jiang N,et al.Molecular basis of rifampicin resistance in multiresistant porcine livestock-associated MRSA[J].J Antimicrob Chemother,2016,71(11):3313-3315.
[13]Cucarella C,Tormo M A,Ubeda C,et al.Role of Biofilm-Associated Protein Bap in the Pathogenesis of Bovine Staphylococcus aureus[J].Infect Immun,2004,72(4):2177-2185.
[14]Bjarnsholt T.The role of bacterial biofilms in chronic infections[J].APMS Suppl,2013,136(5):1-51.
[15]Li T,Lu H,Wang X,et al.Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015[J].Front Cell Infect Microbiol,2017(7):127.
[16]Mohabi S,Kalanta-Nneyestanaki D,Mansouri S.Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by Quercus infectoria gall extracts[J].Iran J Microbiol,2017,9(1):26-32.
[17]Lequette Y,Greenberg E P.Timing and Localization of Rhamnolipid Synthesis Gene Expression in Pseudomonas aeruginosa Biofilms[J].J Bacteriol,2005,187(1):37.
[18]Que Y A,Franε·ois P,Haefliger J A,et al.Reassessing the Role of Staphylococcus aureus Clumping Factor and Fibronectin-Binding Protein by Expression in Lactococcus lactis[J].Infect Immun,2001,69(10):6296.
[19]O'Neill E,Humphreys H,O'Gara J P.Carriage of both the fnb A and fnb B genes and growth at 37 degrees C promote Fn BP-mediated biofilm development in meticillin-resistant Staphylococcus aureus clinical isolates[J].J Med Microbiol,2009,58(4):399-402.
[20]Szczuka E,Urbańska K,Pietryka M,et al.Biofilm density and detection of biofilm-producing genes in methicillin-resistant Staphylococcus aureus strains[J].Folia Microbiol(Praha),2013,58(1):47-52.
[21]Beauregard P B,Chai Y,Vlamakis H,et al.Bacillus subtilis biofilm induction by plant polysaccharides[J].Proc Nati Acad Sci U S A,2013,110(17):1621-1630.
[2]Mcdougall S,Hussein H,Petrovski K.Antimicrobial resistance in Staphylococcus aureus,Streptococcus uberis and Streptococcus dysgalactiae from dairy cows with mastitis[J].New Zealand Veterinary Journal,2014,62(2):68-76.
[3]杨峰,王旭荣,李新圃,等.牛源金黄色葡萄球菌耐药性与相关耐药基因和菌株毒力基因的相关性研究[J].中国兽医科学,2016(2):247-252.
[4]金耀忠,万世平,姜法铭,等.奶牛乳房炎流行情况调查及主要病原分离与药敏试验[J].畜牧与兽医,2011,43(5):64-67.
[5]Serray B,Oufrid S,Hannaoui I,et al.Genes encoding adhesion factors and biofilm formation in methicillin-resistant Staphylococcus aureus in Morocco[J].J Infect Dev Ctries,2016,10(8):863.
[6]Gogoi-Tiwari J,Williams V,Waryah C B,et al.Mammary Gland Pathology Subsequent to Acute Infection with Strong versus Weak Biofilm Forming Staphylococcus aureus Bovine Mastitis Isolates:A Pilot Study Using Non-Invasive Mouse Mastitis Model[J].Plos One,2017,12(1):17-68.
[7]Cha J O,Yoo J I,Yoo J S,et al.Investigation of Biofilm Formation and its Association with the Molecular and Clinical Characteristics of Methicillin-resistant Staphylococcus aureus[J].Osong Public Health Res Perspect,2014,5(2):116.
[8]Nourbakhsh F,Namvar A E.Detection of genes involved in biofilm formation in Staphylococcus aureus isolates[J].GMS Hyg Infect Control,2016,11.
[9]Yang Y,Zhang X,Huang W,et al.SCCmec-associated psm-mec,mRNA promotes Staphylococcus epidermidis,biofilm formation[J].Antonie Van Leeuwenhoek,2016,109(10):1403-1415.
[10]Pereyra E A,Picech F,Renna M S,et al.Detection of Staphylococcus aureus adhesion and biofilm-producing genes and their expression during internalization in bovine mammary epithelial cells[J].Vet Microbiol,2016,183:69-77.
[11]Coelho S M,Reinoso E,Pereira I A,et al.Virulence factors and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Rio de Janeiro[J].Pesquisa Veterinária Brasileira,2009,29(5):369-374.
[12]Li J,Feβler A T,Jiang N,et al.Molecular basis of rifampicin resistance in multiresistant porcine livestock-associated MRSA[J].J Antimicrob Chemother,2016,71(11):3313-3315.
[13]Cucarella C,Tormo M A,Ubeda C,et al.Role of Biofilm-Associated Protein Bap in the Pathogenesis of Bovine Staphylococcus aureus[J].Infect Immun,2004,72(4):2177-2185.
[14]Bjarnsholt T.The role of bacterial biofilms in chronic infections[J].APMS Suppl,2013,136(5):1-51.
[15]Li T,Lu H,Wang X,et al.Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015[J].Front Cell Infect Microbiol,2017(7):127.
[16]Mohabi S,Kalanta-Nneyestanaki D,Mansouri S.Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by Quercus infectoria gall extracts[J].Iran J Microbiol,2017,9(1):26-32.
[17]Lequette Y,Greenberg E P.Timing and Localization of Rhamnolipid Synthesis Gene Expression in Pseudomonas aeruginosa Biofilms[J].J Bacteriol,2005,187(1):37.
[18]Que Y A,Franε·ois P,Haefliger J A,et al.Reassessing the Role of Staphylococcus aureus Clumping Factor and Fibronectin-Binding Protein by Expression in Lactococcus lactis[J].Infect Immun,2001,69(10):6296.
[19]O'Neill E,Humphreys H,O'Gara J P.Carriage of both the fnb A and fnb B genes and growth at 37 degrees C promote Fn BP-mediated biofilm development in meticillin-resistant Staphylococcus aureus clinical isolates[J].J Med Microbiol,2009,58(4):399-402.
[20]Szczuka E,Urbańska K,Pietryka M,et al.Biofilm density and detection of biofilm-producing genes in methicillin-resistant Staphylococcus aureus strains[J].Folia Microbiol(Praha),2013,58(1):47-52.
[21]Beauregard P B,Chai Y,Vlamakis H,et al.Bacillus subtilis biofilm induction by plant polysaccharides[J].Proc Nati Acad Sci U S A,2013,110(17):1621-1630.