阿萨希毛孢子菌胞外分泌酶和药物敏感性的微进化研究
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摘要
目的研究宿主体内微进化对阿萨希毛孢子菌(Trichosporon asahii,T.asahii)胞外分泌酶和三唑类抗真菌药物敏感性的影响。方法酶检测培养基分析菌株胞外分泌酶种类和活性,采用美国临床实验室标准化研究所(CLSI)制定的M27-A3实验方案进行药物敏感实验。结果原代株(TO)胞外分泌酶谱组成及活性:蛋白酶(++++)、脂肪酶(++)、溶血素(++++)和磷脂酶(+++),微进化株(TEVO)胞外分泌酶谱组成及活性:蛋白酶(+)、脂肪酶(++)、溶血素(+++)。原代株的氟康唑、伊曲康唑和伏立康唑MIC值分别为8μg/ml、0.5μg/ml和0.125μg/ml,微进化株的MIC值分别为32μg/ml、4μg/ml、0.25μg/ml。结论在宿主内环境和抗真菌药物双重压力下,阿萨希毛孢子菌改变其胞外分泌酶谱、降低部分毒力并增强耐药性以适应宿主内环境,从而与宿主形成长期共存状态。
OBJECTIVE To study the effects of microevolution on the extracellular enzyme and drug susceptibility of Trichosporon asahii(T.asahii).METHODS The species and activity of the extracellular enzymes were analyzed by using enzyme assay media,and the drug susceptibility testing was performed with the use of the M27-A3 protocol by American Clinical and Laboratory Standards Institute(CLSI).RESULTS The extracellular enzyme profile and activity of original strain(TO)included:protease(+ + + +),lipase(+ +),hemolysin(+ + + +)and phospholipase(+ + +).The extracellular enzyme profile and activity of evolutional strain(TEVO)included:protease(+),lipase(+ +)and hemolysin(+ + +).The original strain showed a minimal inhibitory concentration(MIC)of 8μg/ml for fluconazole,0.5μg/ml for itraconazole and 0.125μg/ml for voriconazole,however,the evolutional strain showed a MIC of 32μg/ml for fluconazole,4μg/ml for itraconazole and 0.5μg/ml for voriconazole.CONCLUSION Under the dual pressure from host environment and antifungal therapy,T.asahii change the profile of extracellular enzyme,degrade some kinds of virulence factors and increase resistance to antifungal agents,and these microevolution manifestations of T.asahii contribute to its adaption to host environment and coexistence with host.
OBJECTIVE To study the effects of microevolution on the extracellular enzyme and drug susceptibility of Trichosporon asahii(T.asahii).METHODS The species and activity of the extracellular enzymes were analyzed by using enzyme assay media,and the drug susceptibility testing was performed with the use of the M27-A3 protocol by American Clinical and Laboratory Standards Institute(CLSI).RESULTS The extracellular enzyme profile and activity of original strain(TO)included:protease(+ + + +),lipase(+ +),hemolysin(+ + + +)and phospholipase(+ + +).The extracellular enzyme profile and activity of evolutional strain(TEVO)included:protease(+),lipase(+ +)and hemolysin(+ + +).The original strain showed a minimal inhibitory concentration(MIC)of 8μg/ml for fluconazole,0.5μg/ml for itraconazole and 0.125μg/ml for voriconazole,however,the evolutional strain showed a MIC of 32μg/ml for fluconazole,4μg/ml for itraconazole and 0.5μg/ml for voriconazole.CONCLUSION Under the dual pressure from host environment and antifungal therapy,T.asahii change the profile of extracellular enzyme,degrade some kinds of virulence factors and increase resistance to antifungal agents,and these microevolution manifestations of T.asahii contribute to its adaption to host environment and coexistence with host.
引文
[1]Gouba N,Raoult D,Drancourt M.Eukaryote culturomics of the gut reveals new species[J].PLoS One,2014,9(9):e106994.
[2]Mada PK,Ayoade F,Li A,et al.Trichosporon asahii septic thrombophlebitis following lower extremity amputation in an immunocompetent host[J].BMJ Case Rep,2018,2018.pii:bcr-2017-221441.
[3]Valle RS,Ramos LS,Reis VJ,et al.Trichosporon asahii secretes a 30-kDa aspartic peptidase[J].Microbiol Res,2017,205:66-72.
[4]Schaller M,Borelli C,Korting HC,et al.Hydrolytic enzymes as virulence factors of Candida albicans[J].Mycoses,2005,48(6):365-377.
[5]de Paula MR,de Melo RE,Borges AS,et al.Evaluation of virulence factors of Candida albicans isolated from HIV-positive individuals using HAART[J].Arch Oral Biol,2016,66:61-65.
[6]Arendrup MC,Boekhout T,Akova M,et al.ESCMID and ECMMjoint clinical guidelines for the diagnosis and management of rare invasive yeast infections[J].Clin Microbiol Infect,2014,20(Suppl3):76-98.
[7]Liao Y,Lu X,Yang S,et al.Epidemiology and outcome of Trichosporon fungemia:a review of 185reported cases from1975to 2014[J].Open Forum Infect Dis,2015,2(4):v141.
[8]杨蓉娅,敖俊红,王文岭,等.阿萨希丝孢酵母引起播散性毛孢子菌病国内首例报告[J].中华皮肤科杂志,2001,34(5):329-332.
[9]Canela H,Cardoso B,Vitali LH,et al.Prevalence,virulence factors and antifungal susceptibility of Candida spp.isolated from bloodstream infections in a tertiary care hospital in Brazil[J].Mycoses,2018,61(1):11-21.
[10]陈淑贤,陈惠玲,叶飘,等.医院感染假丝酵母菌病磷脂酶毒力与溶血性毒力的分析[J].中华医院感染学杂志,2016,26(2):274-276.
[11]Elavarashi E,Kindo AJ,Rangarajan S.Enzymatic and nonenzymatic virulence activities of dermatophytes on solid media[J].J Clin Diagn Res,2017,11(2):C23-C25.
[12]Pumipuntu N,Kulpeanprasit S,Santajit S,et al.Screening method for Staphylococcus aureus identification in subclinical bovine mastitis from dairy farms[J].Vet World,2017,10(7):721-726.
[13]Clinical and Laboratory Standards Institute.Referencemethod for broth dilution antifungal susceptibility testing ofyeasts[S].Approved standard M27-A3[S].Wayne,USA:CLSI,2008.
[14]Li H,Wang C,Chen Y,et al.Integrated transcriptomic analysis of Trichosporon asahii uncovers the core genes and pathways of fluconazole resistance[J].Sci Rep,2017,7(1):17847.
[15]Bentubo HD,Gompertz OF.Effects of temperature and incubation time on the in vitro expression of proteases,phospholipases,lipases and DNases by different species of Trichosporon[J].Springerplus,2014,3:377.
[16]Sun W,Su J,Xu S,et al.Trichosporon asahii causing nosocomial urinary tract infections in intensive care unit patients:genotypes,virulence factors and antifungal susceptibility testing[J].J Med Microbiol,2012,61(Pt 12):1750-1757.
[17]Hazirolan G,Kocak N,Karagoz A.Sequence-based identification,genotyping and virulence factors of Trichosporon asahii strains isolated from urine samples of hospitalized patients(2011-2016)[J].J Mycol Med,2018,28(3):452-456.
[18]Montoya AM,Sanchez GA,Palma-Nicolas JP,et al.Genotyping,extracellular compounds,and antifungal susceptibility testing of Trichosporon asahii isolated from Mexican patients[J].Med Mycol,2015,53(5):505-511.
[19]Favero D,Furlaneto-Maia L,Franca EJ,et al.Hemolytic factor production by clinical isolates of Candida species[J].Curr Microbiol,2014,68(2):161-166.
[20]Pfaller MA,Diekema DJ,Gibbs DL,et al.Results from the ARTEMIS DISK global antifungal surveillance study,1997to2007:10.5-year analysis of susceptibilities of noncandidal yeast species to fluconazole and voriconazole determined by CLSI standardized disk diffusion testing[J].J Clin Microbiol,2009,47(1):117-123.
[21]de Almeida JJ,Hennequin C.Invasive Trichosporon infection:a systematic review on a re-emerging fungal pathogen[J].Front Microbiol,2016,7:1629.
[22]郭朝霞,李海涛,祝贺,等.不同来源阿萨希毛孢子菌体外药物敏感性及致病性研究[J].中华医院感染学杂志,2013,23(24):5915-5917.
[23]Falk R,Wolf DG,Shapiro M,et al.Multidrug-resistant Trichosporon asahii isolates are susceptible to voriconazole[J].JClin Microbiol,2003,41(2):911.
[2]Mada PK,Ayoade F,Li A,et al.Trichosporon asahii septic thrombophlebitis following lower extremity amputation in an immunocompetent host[J].BMJ Case Rep,2018,2018.pii:bcr-2017-221441.
[3]Valle RS,Ramos LS,Reis VJ,et al.Trichosporon asahii secretes a 30-kDa aspartic peptidase[J].Microbiol Res,2017,205:66-72.
[4]Schaller M,Borelli C,Korting HC,et al.Hydrolytic enzymes as virulence factors of Candida albicans[J].Mycoses,2005,48(6):365-377.
[5]de Paula MR,de Melo RE,Borges AS,et al.Evaluation of virulence factors of Candida albicans isolated from HIV-positive individuals using HAART[J].Arch Oral Biol,2016,66:61-65.
[6]Arendrup MC,Boekhout T,Akova M,et al.ESCMID and ECMMjoint clinical guidelines for the diagnosis and management of rare invasive yeast infections[J].Clin Microbiol Infect,2014,20(Suppl3):76-98.
[7]Liao Y,Lu X,Yang S,et al.Epidemiology and outcome of Trichosporon fungemia:a review of 185reported cases from1975to 2014[J].Open Forum Infect Dis,2015,2(4):v141.
[8]杨蓉娅,敖俊红,王文岭,等.阿萨希丝孢酵母引起播散性毛孢子菌病国内首例报告[J].中华皮肤科杂志,2001,34(5):329-332.
[9]Canela H,Cardoso B,Vitali LH,et al.Prevalence,virulence factors and antifungal susceptibility of Candida spp.isolated from bloodstream infections in a tertiary care hospital in Brazil[J].Mycoses,2018,61(1):11-21.
[10]陈淑贤,陈惠玲,叶飘,等.医院感染假丝酵母菌病磷脂酶毒力与溶血性毒力的分析[J].中华医院感染学杂志,2016,26(2):274-276.
[11]Elavarashi E,Kindo AJ,Rangarajan S.Enzymatic and nonenzymatic virulence activities of dermatophytes on solid media[J].J Clin Diagn Res,2017,11(2):C23-C25.
[12]Pumipuntu N,Kulpeanprasit S,Santajit S,et al.Screening method for Staphylococcus aureus identification in subclinical bovine mastitis from dairy farms[J].Vet World,2017,10(7):721-726.
[13]Clinical and Laboratory Standards Institute.Referencemethod for broth dilution antifungal susceptibility testing ofyeasts[S].Approved standard M27-A3[S].Wayne,USA:CLSI,2008.
[14]Li H,Wang C,Chen Y,et al.Integrated transcriptomic analysis of Trichosporon asahii uncovers the core genes and pathways of fluconazole resistance[J].Sci Rep,2017,7(1):17847.
[15]Bentubo HD,Gompertz OF.Effects of temperature and incubation time on the in vitro expression of proteases,phospholipases,lipases and DNases by different species of Trichosporon[J].Springerplus,2014,3:377.
[16]Sun W,Su J,Xu S,et al.Trichosporon asahii causing nosocomial urinary tract infections in intensive care unit patients:genotypes,virulence factors and antifungal susceptibility testing[J].J Med Microbiol,2012,61(Pt 12):1750-1757.
[17]Hazirolan G,Kocak N,Karagoz A.Sequence-based identification,genotyping and virulence factors of Trichosporon asahii strains isolated from urine samples of hospitalized patients(2011-2016)[J].J Mycol Med,2018,28(3):452-456.
[18]Montoya AM,Sanchez GA,Palma-Nicolas JP,et al.Genotyping,extracellular compounds,and antifungal susceptibility testing of Trichosporon asahii isolated from Mexican patients[J].Med Mycol,2015,53(5):505-511.
[19]Favero D,Furlaneto-Maia L,Franca EJ,et al.Hemolytic factor production by clinical isolates of Candida species[J].Curr Microbiol,2014,68(2):161-166.
[20]Pfaller MA,Diekema DJ,Gibbs DL,et al.Results from the ARTEMIS DISK global antifungal surveillance study,1997to2007:10.5-year analysis of susceptibilities of noncandidal yeast species to fluconazole and voriconazole determined by CLSI standardized disk diffusion testing[J].J Clin Microbiol,2009,47(1):117-123.
[21]de Almeida JJ,Hennequin C.Invasive Trichosporon infection:a systematic review on a re-emerging fungal pathogen[J].Front Microbiol,2016,7:1629.
[22]郭朝霞,李海涛,祝贺,等.不同来源阿萨希毛孢子菌体外药物敏感性及致病性研究[J].中华医院感染学杂志,2013,23(24):5915-5917.
[23]Falk R,Wolf DG,Shapiro M,et al.Multidrug-resistant Trichosporon asahii isolates are susceptible to voriconazole[J].JClin Microbiol,2003,41(2):911.