山羊源精氨酸支原体的分离鉴定与药感试验
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摘要
本实验的目的是从患有呼吸道疾病的山羊中分离支原体。采集13个病羊鼻腔棉拭子,接种于支原体液体培养基中,37℃、5%CO2培养6d,采用16Sr RNA通用引物检测支原体属PCR方法进行检测,阳性样本转移固体培养基分离鉴定并进行药敏试验。结果显示,从13份液体培养基中检测出5株支原体,经16Sr DNA序列分析证实为精氨酸支原体,遗传进化分析表明,5株精氨酸支原体独立的聚为1支。在固体培养基上分离得到2株支原体,药敏试验结果显示对红霉素、盐酸土霉素、酒石酸泰乐菌素、氧氟沙星、硫酸头孢喹诺、恩诺沙星、林可霉素、硫酸庆大霉素、头孢噻呋、硫酸链霉素10种抗生素敏感,对氟苯尼考耐药。本实验从患呼吸道病的山羊中成功分离出精氨酸支原体,鉴于该菌是人的致病菌,其公共卫生学意义值得进一步关注。
The purpose of this study was to isolate mycoplasma from goats with respiratory disease. 13 nasal swab samples were collected from diseased goats and inoculated in the liquid medium,and cultures were incubated for six days at 37℃ 、5%CO_2. The samples were detected by a PCR for specific detecting mycoplasma,and the positive samples were transferred to solid medium for further drug sensitivity test. The results showed that 5 positives were detected as mycoplasma positive from 13 samples,and were identificated as mycoplasma arginini by 16 Sr RNA sequences. Genetic evolution analysis based on 16 Sr RNA sequences showed that 5 strains of 16 Sr RNA sequences clustered into one independent branches. 2 strains of mycoplasma arginini were isolated using solid medium. The results of drug sensitivity test showed that the strains were sensitive to erythromycin,Oxytetracycline hydrochloride,tartaric acid tylosin,ofloxacin,cefquinome sulfate,enrofloxacin,clindamycin,gentamycin sulfate,ceftiofur, streptomycin sulfate and resistance to florfenicol. As mycoplasma arginini is a pathogen to human,the public health significance of mycoplasma arginini in goats deserves further attention.
The purpose of this study was to isolate mycoplasma from goats with respiratory disease. 13 nasal swab samples were collected from diseased goats and inoculated in the liquid medium,and cultures were incubated for six days at 37℃ 、5%CO_2. The samples were detected by a PCR for specific detecting mycoplasma,and the positive samples were transferred to solid medium for further drug sensitivity test. The results showed that 5 positives were detected as mycoplasma positive from 13 samples,and were identificated as mycoplasma arginini by 16 Sr RNA sequences. Genetic evolution analysis based on 16 Sr RNA sequences showed that 5 strains of 16 Sr RNA sequences clustered into one independent branches. 2 strains of mycoplasma arginini were isolated using solid medium. The results of drug sensitivity test showed that the strains were sensitive to erythromycin,Oxytetracycline hydrochloride,tartaric acid tylosin,ofloxacin,cefquinome sulfate,enrofloxacin,clindamycin,gentamycin sulfate,ceftiofur, streptomycin sulfate and resistance to florfenicol. As mycoplasma arginini is a pathogen to human,the public health significance of mycoplasma arginini in goats deserves further attention.
引文
[1]Rose D L,Tully J G.Fatal Septicemia Due to Mycoplasma arginini:A New Human Zoonosis[J].Clinical Infectious Diseases,1992,15(3):434-438.
[2]Yechouron A,Lefebvre J,Robson H G,et al.Fatal septicemia due to Mycoplasma arginini:a new human zoonosis[J].Clinical Infectious Diseases,1992,15(3):434-438.
[3]Watanabe M,Hitomi S,Goto M,et al.Bloodstream infection due to Mycoplasma arginini in an immunocompromised patient.[J].Journal of Clinical Microbiology,2012,50(9).
[4]SillóP,Pintér D,Ostorházi E,et al.Eosinophilic Fasciitis Associated with Mycoplasma arginini Infection[J].Journal of Clinical Microbiology,2012,50(3).
[5]Prayson M J,Venkatarayappa I,Srivastava M,et al.Deep infection with Mycoplasma arginini,in an open femur fracture secondary to an African lion bite:A case report[J].Injury Extra,2008.
[6]Tan R J,Miles J A.Incidence and significance of mycoplasmas in sick cats.[J].Research in Veterinary Science,1974,16(1):27-34.39(7):243-246.
[7]Kusiluka L J M,Ojeniyi B,Friis N F,et al.Mycoplasmas isolated from the respiratory tract of cattle and goats in Tanzania.[J].Acta Veterinaria Scandinavica,2000,41(3):299.
[8]Akwuobu C A,Ayling R D,Chah K F,et al.Studies into the prevalence of Mycoplasma species in small ruminants in Benue State,North-central Nigeria.[J].Tropical Animal Health&Production,2014,46(6):1087-1092.
[9]Fern2n S,Galapero J,Rey J,et al.Investigations into the seasonal presence of Mycoplasma species in fattening lambs[J].Veterinary Journal,2016,(212):80-82.
[10]Alaubaidi J M,Taylor W D,Bubash G R,et al.Identification and characterization of Mycoplasma arginini from Bighorn sheep(Ovis canadensis)and goats.[J].American Journal of Veterinary Research,1972,33(1):87-90.
[11]Elfaki M G,Abbas B,Mahmoud O M,et al.Isolation and characterization of Mycoplasma arginini,from camels(Camelus dromedarius)with pneumonia[J].Comparative Immunology Microbiology&Infectious Diseases,2002,25(1):49.
[12]Bottinelli M,Schnee C,Lepri E,et al.Investigation on mycoplasma populations in pneumonic dairy lamb lungs using a DNA microarray assay[J].Small Ruminant Research,2017,(147):96-100.
[13]Elshafey D,Awad N,Elnaker Y F,et al.Isolation and Molecular Characterization of Mycolplasm arginini isolated from sheep and goats with respiratory manifestation in Dakahilia Governorate[C]//Conference of the Scientific Research Institute of Animal Society modern Trends Health for the Advancement of Animal Health.2016.
[14]Valsala R,Rana R,Remesh A T,et al.Mycoplasma arginini:High frequency involvement in goat pneumonia[J].Turkish Journal of Veterinary&Animal Sciences,2017,41(3):393-399.
[15]Gon9alves R,Mariano I,Nú1ez A,et al.Atypical nonprogressive pneumonia in goats[J].Veterinary Journal,2010,183(2):219.
[16]王栋,刘众心,张立春.从山羊分离出绵羊肺炎霉形体的报告[J].中国兽医杂志,1991,(6).
[17]郑佳琪,黄海碧,王晓晖,等.绵羊肺炎支原体、丝状支原体山羊亚种和精氨酸支原体多重PCR检测方法的建立及应用[J].中国兽医学报,2016,36(7):1131-1134.
[18]江锦秀,林裕胜,游伟,等.福建省羊支原体性肺炎分子流行病学研究[J].福建农业学报,2017,32(1):12-16.
[19]Rawadi G,Dujeancourt-Henry A,Lemercier B,et al.Phylogenetic position of rare human mycoplasmas,Mycoplasma faucium,M.buccale,M.primatum and M.spermatophilum,based on 16S r RNA gene sequences[J].Int J Syst Bacteriol,1998,48 Pt 1(1):305-309.
[20]Hata E.Complete Genome Sequence of Mycoplasma arginini Strain HAZ 145_1 from Bovine Mastitic Milk in Japan.[J].Genome Announcements,2014,2(5).
[21]Olaogun O M,Kanci A,Barber S R,et al.Genetic diversity of Mycoplasma arginini isolates based on multilocus sequence typing[J].Veterinary Microbiology,2015,180(1-2):123-128.
[22]张莉,王作艳,孙继国,等.绵羊肺炎支原体的微生物学特性的研究[J].河北农业大学学报,2003,26(z1):221-224.
[23]Elfaki M G,Abbas B,Mahmoud O M,et al.Isolation and characterization of Mycoplasma arginini from camels(Camelus dromedarius)with pneumonia[J].Comparative Immunology Microbiology&Infectious Diseases,2002,25(1):49-57.
[2]Yechouron A,Lefebvre J,Robson H G,et al.Fatal septicemia due to Mycoplasma arginini:a new human zoonosis[J].Clinical Infectious Diseases,1992,15(3):434-438.
[3]Watanabe M,Hitomi S,Goto M,et al.Bloodstream infection due to Mycoplasma arginini in an immunocompromised patient.[J].Journal of Clinical Microbiology,2012,50(9).
[4]SillóP,Pintér D,Ostorházi E,et al.Eosinophilic Fasciitis Associated with Mycoplasma arginini Infection[J].Journal of Clinical Microbiology,2012,50(3).
[5]Prayson M J,Venkatarayappa I,Srivastava M,et al.Deep infection with Mycoplasma arginini,in an open femur fracture secondary to an African lion bite:A case report[J].Injury Extra,2008.
[6]Tan R J,Miles J A.Incidence and significance of mycoplasmas in sick cats.[J].Research in Veterinary Science,1974,16(1):27-34.39(7):243-246.
[7]Kusiluka L J M,Ojeniyi B,Friis N F,et al.Mycoplasmas isolated from the respiratory tract of cattle and goats in Tanzania.[J].Acta Veterinaria Scandinavica,2000,41(3):299.
[8]Akwuobu C A,Ayling R D,Chah K F,et al.Studies into the prevalence of Mycoplasma species in small ruminants in Benue State,North-central Nigeria.[J].Tropical Animal Health&Production,2014,46(6):1087-1092.
[9]Fern2n S,Galapero J,Rey J,et al.Investigations into the seasonal presence of Mycoplasma species in fattening lambs[J].Veterinary Journal,2016,(212):80-82.
[10]Alaubaidi J M,Taylor W D,Bubash G R,et al.Identification and characterization of Mycoplasma arginini from Bighorn sheep(Ovis canadensis)and goats.[J].American Journal of Veterinary Research,1972,33(1):87-90.
[11]Elfaki M G,Abbas B,Mahmoud O M,et al.Isolation and characterization of Mycoplasma arginini,from camels(Camelus dromedarius)with pneumonia[J].Comparative Immunology Microbiology&Infectious Diseases,2002,25(1):49.
[12]Bottinelli M,Schnee C,Lepri E,et al.Investigation on mycoplasma populations in pneumonic dairy lamb lungs using a DNA microarray assay[J].Small Ruminant Research,2017,(147):96-100.
[13]Elshafey D,Awad N,Elnaker Y F,et al.Isolation and Molecular Characterization of Mycolplasm arginini isolated from sheep and goats with respiratory manifestation in Dakahilia Governorate[C]//Conference of the Scientific Research Institute of Animal Society modern Trends Health for the Advancement of Animal Health.2016.
[14]Valsala R,Rana R,Remesh A T,et al.Mycoplasma arginini:High frequency involvement in goat pneumonia[J].Turkish Journal of Veterinary&Animal Sciences,2017,41(3):393-399.
[15]Gon9alves R,Mariano I,Nú1ez A,et al.Atypical nonprogressive pneumonia in goats[J].Veterinary Journal,2010,183(2):219.
[16]王栋,刘众心,张立春.从山羊分离出绵羊肺炎霉形体的报告[J].中国兽医杂志,1991,(6).
[17]郑佳琪,黄海碧,王晓晖,等.绵羊肺炎支原体、丝状支原体山羊亚种和精氨酸支原体多重PCR检测方法的建立及应用[J].中国兽医学报,2016,36(7):1131-1134.
[18]江锦秀,林裕胜,游伟,等.福建省羊支原体性肺炎分子流行病学研究[J].福建农业学报,2017,32(1):12-16.
[19]Rawadi G,Dujeancourt-Henry A,Lemercier B,et al.Phylogenetic position of rare human mycoplasmas,Mycoplasma faucium,M.buccale,M.primatum and M.spermatophilum,based on 16S r RNA gene sequences[J].Int J Syst Bacteriol,1998,48 Pt 1(1):305-309.
[20]Hata E.Complete Genome Sequence of Mycoplasma arginini Strain HAZ 145_1 from Bovine Mastitic Milk in Japan.[J].Genome Announcements,2014,2(5).
[21]Olaogun O M,Kanci A,Barber S R,et al.Genetic diversity of Mycoplasma arginini isolates based on multilocus sequence typing[J].Veterinary Microbiology,2015,180(1-2):123-128.
[22]张莉,王作艳,孙继国,等.绵羊肺炎支原体的微生物学特性的研究[J].河北农业大学学报,2003,26(z1):221-224.
[23]Elfaki M G,Abbas B,Mahmoud O M,et al.Isolation and characterization of Mycoplasma arginini from camels(Camelus dromedarius)with pneumonia[J].Comparative Immunology Microbiology&Infectious Diseases,2002,25(1):49-57.