牛支原体新疆株的分离鉴定

doi:10.16431/j.cnki.1671-7236.2019.05.025

Abstract

Abstract:

In order to investigate and determine the pathogen from calf which showed pneumonia symptoms occurred in cattle farms in Xinjiang,one strain of Mycoplasma bovis was isolated from seven pneumonic lung tissue samples of calves using liquid and solid Mycoplasma medium.The isolated strain was characterized by the colonial morphologic observation and biochemistry test.Specific primers of Mycoplasma bovis and universal 16S rRNA primers of Mycoplasma were employed to amplify and the amplicons were sequenced as well.DNAStar software was used to compare the sequence between the isolate and the other strains deposited in GenBank.A phylogenetic tree of different strains based on 16S rRNA sequences was constructed by Mega 6.0 software with the Neighbor-Joining method.The results showed that colonies of isolated strain grew on solid medium were typical fried-eggs,the central depression of the colony was deep into the medium and thin and transparent periphery,and Dienes staining was dark blue.The isolated strain could not ferment glucose,not decompose urea,and not hydrolyze arginine.Hemadsorption test and hemolysis test were negative,while film and spot test and triphenyl tetrazolium chloride reduction reaction test were positive.PCR products of Mycoplasma bovis showed the specific band of 1 911 bp.Sequence analysis indicated that 16S rRNA of the isolated strain showed 99.8% homology with that of Mycoplasma bovis strain PG45,and 99.3% to 99.7% homology with that of local strains of Mycoplasma bovis (Mb NM2012,Mb HB0801,Mb Hubei-1,Mb Ningxia-1,Mb CQ-W70 and Mb 08M strains).The phylogenetic tree showed that the 16S rRNA gene of the isolated strain was closely relative with Mb Ningxia-1 and Mb 08M strains,which were in the same branch.The results confirmed that the pathogenic bacteria causing cattle death was Mycoplasma bovis,which provided theoretical basis and scientific basis for the prevention and treatment of Mycoplasma bovis in Xinjiang.

Key words: Mycoplasma bovis; isolation; identification; 16S rRNA

CLC Number:

S852.62

LING Chen, HAO Chengwu, HE Hai, ZHANG Fei, HOU Feng, HE Sun. Isolation and Identification of Mycoplasma bovis in Xinjiang[J]. , 2019, 46(5): 1466-1473.

References

[1] NICHOLAS R A,AYLING R D.Mycoplasma bovis:Disease,diagnosis and control[J].Research in Veterinary Science,2003,74(2):105-112.
[2] 季文恒,陈胜利,赵萍,等.牛支原体致病机理的研究进展[J].中国预防兽医学报,2018,40(4):361-367. JI W H,CHEN S L,ZHAO P,et al.Progress in the pathogenesis of Mycoplasma bovis[J]. Chinese Journal of Preventive Veterinary Medicine,2018,40(4):361-367.(in Chinese)
[3] 陆承平.兽医微生物学[M].第3版.北京:中国农业出版社,2001. LU C P.Veterinary Microbiology[M].3rd Edition.Beijing:China Agriculture Press,2001.(in Chinese)
[4] 杨铭伟.牛支原体灭活疫苗的研制与免疫效果研究[D].石河子:石河子大学,2015. YANG M W.Research on the development of Mycoplasma bovis inactivated vaccine and immune effect in calves[D].Shihezi:Shihezi University,2015.(in Chinese)
[5] 黎济申,陈嘉棣,王兆久,等.牛霉形体的分离及其对犊牛的致病力[J].畜牧与兽医,1984,1:8-9. LI J S,CHEN J D,WANG Z J,et al.Isolation of Mycoplasma bovis and its pathogenicity to calves[J].Animal Husbandry & Veterinary Medicine,1984,1:8-9.(in Chinese)
[6] 石磊,龚瑞,尹争艳,等.肉牛传染性牛支原体肺炎流行的诊断[J].华中农业大学学报,2008,27(4):629-633. SHI L,GONG R,YIN Z Y,et al.Diagnosis of cattle infectious Mycoplasma bovis pneumonia[J].Journal of Huazhong Agricultural University,2008,27(4):629-633.(in Chinese)
[7] 蒋南,翟研妮,谷长勤,等.传染性牛支原体肺炎自然感染病牛的病理学观察[J].中国兽医科学,2011,41(9):933-935. JIANG N,ZHAI Y N,GU C Q,et al.Pathological observation of infectious Mycoplasma bovis pneumonia in nature infection[J].Chinese Veterinary Science,2011,41(9):933-935.(in Chinese)
[8] 辛九庆,李媛,郭丹,等.国内首次从患肺炎的犊牛肺脏中分离到牛支原体[J].中国预防兽医学报,2008,30(9):661-664. XIN J Q,LI Y,GUO D,et al.First isolation of Mycoplasma bovis from calf lung with pneumoniae in China[J].Chinese Journal of Preventive Veterinary Medicine,2008,30(9):661-664.(in Chinese)
[9] 姚永进,剡根强,王静梅,等.致犊牛肺炎和关节炎牛支原体新疆株的分离与鉴定[J].中国畜牧兽医,2011,38(12):76-79. YAO Y J,YAN G Q,WANG J M,et al.Isolation and identification of Mycoplasma bovis Xinjiang strain inducing pneumonia and arthritis of calf[J].China Animal Husbandry & Veterinary Medicine,2011,38(12):76-79.(in Chinese)
[10] 季文恒,吴娅琴,翟肖辉,等.牛支原体疫苗的研究进展[J].中国畜牧兽医,2018,45(3):763-769. JI W H,WU Y Q,ZHAI X H,et al.Advances on the research of vaccine against Mycoplasma bovis[J].China Animal Husbandry & Veterinary Medicine,2018,45(3):763-769.(in Chinese)
[11] DEDIEU L,MADY V,LEFEVRE P C.Development of a selective polymerase chain reaction assay for the detection of Mycoplasma mycoides subsp. Mycoides S.C.(contagious bovine pleuropneumonia agent)[J].Veterinary Microbiology,1994,42(4):327-339.
[12] 冉智光,谢建华,骆璐,等.我国部分地区牛支原体肺炎和关节炎的病原体诊断[J].中国预防兽医学报,2010,32(1):40-43. RAN Z G,XIE J H,LUO L,et al.Etiological diagnosis of Mycoplasma bovis pneumonia and arthritis in some provinces in China[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(1):40-43.(in Chinese)
[13] 杨铭伟,李迪,赵海龙,等.致犊牛肺炎牛支原体南疆株的分离鉴定及oppF基因序列分析[J].中国畜牧兽医,2014,41(12):25-29. YANG M W,LI D,ZHAO H L,et al.Isolation and identification of Mycoplasma bovis Nanjiang strain inducing pneumonia and sequence analysis of oppF gene[J].China Animal Husbandry & Veterinary Medicine,2014,41(12):25-29.(in Chinese)
[14] 吴移谋,叶元康.支原体学[M].第2版.北京:人民卫生出版社,2008. WU Y M,YE Y K.Mycoplasmology[M].2nd Edition.Beijing:People's Medical Press,2008.(in Chinese)
[15] 师燕霞,凌晨,杨铭伟,等.新疆不同地区牛支原体分离株oppD/F基因的克隆和序列分析[J].中国兽医学报,2018,38(2):311-315. SHI Y X,LING C,YANG M W,et al.Cloning and sequencing of oppD/F gene of Mycoplasma bovis isolated from different regions of Xinjiang[J].Chinese Journal of Veterinary Science,2018,38(2):311-315.(in Chinese)
[16] 江锦秀,林裕胜,游伟,等.牛支原体FJ-HJ分离株的分离及鉴定[J].中国农学通报,2017,33(20):107-112. JIANG J X,LIN Y S,YOU W,et al.Isolation and identification of Mycoplasma bovis FJ-HJ strain[J].Chinese Agricultural Science Bulletin,2017,33(20):107-112.(in Chinese)
[17] RIFATBEGOVIC M,ASSUNÇÃO P,POVEDA J B, et al.Isolation of Mycoplasma bovis from the respiratory tract of cattle in Bosnia and Herzegovina[J].The Veterinary Record,2007,160(14):484-485.
[18] SUBRAMANIAM S,BERGONIER D,POUMARAT F,et al.Species identification of Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR[J].Molecular and Cellular Probes,1998,12(3):161-169.
[19] 李大伟,黄灿平,张彦明,等.牛支原体、无乳支原体和丝状支原体丝状亚种小克隆三重PCR检测方法的建立及初步应用[J].畜牧兽医学报,2011,42(2):306-310. LI D W,HUANG C P,ZHANG Y M,et al.Establishment and application of the triplex PCR for detecting M.bovis,M.mycoides subsp.mycoides small colony type and M.agalactiae[J].Acta Veterinaria et Zootechnica Sinica,2011,42(2):306-310.(in Chinese)
[20] 石磊,龚瑞,尹争艳,等.肉牛传染性牛支原体肺炎流行的初步诊断[J].华中农业大学学报,2008,27(4):572. SHI L,GONG R,YIN Z Y,et al.Preliminary diagnosis of cattle infectious Mycoplasma bovis pneumonia[J].Journal of Huazhong Agricultural University,2008,27(5):572.(in Chinese)
[21] HALE H H,HELMBOLDT C F,PLASTRIDGE W N,et al.Bovine mastitis caused by a Mycoplasma species[J].The Cornell Veterinarian,1962,52:582-591.
[22] KIRK J H,GLENN K,RUIZ L,et al.Epidemiologic analysis of Mycoplasma spp isolated from bulk-tank milk samples obtained from dairy herds that were members of a milk cooperative[J].Journal of the American Veterinary Medical Association,1997,211(8):1036-1038.
[23] NICHOLAS R A,AYLING R D,STIPKOVITS LASZLO P.An experimental vaccine for calf pneumonia caused by Mycoplasma bovis:Clinical,cultural,serological and pathological findings[J].Vaccine,2002,20(29-30):3569-3575.
[24] BYRNE W J,BALL H J,BRICE N,et al.Application of an indirect ELISA to milk samples to identify cows with Mycoplasma bovis mastitis[J].Veterinary Record,2000,146(13):368-369.
[25] AYLING R D,NICHOLAS R A,JOHANSSON K E.Application of the polymerase chain reaction for the routine identification of Mycoplasma bovis[J].The Veterinary Record,1997,141(12):307-308.
[26] HOTZEL H,SACHSE K,PFVTZNER H.Rapid detection of Mycoplasma bovis in milk samples and nasal swabs using the polymerase chain reaction[J].The Journal of Applied Bacteriology,1996,80(5):505-510.

Isolation and Identification of Mycoplasma bovis in Xinjiang

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LU Yukui, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Isolation,Identification and Pathogenicity Analysis of Avibacterium paragallinarum from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2562-2572.
[2]	PENG Yangxiang, YANG Zhenbiao, YAN Kuiyou, WANG Han, HU De, WANG Zun, LIU Ning, ZHAO Liansheng. From Artificial to Intelligent:Research Progress of Individual Idendification Technology for Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1855-1866.
[3]	LI Zhijie, ZHAO Ying, MA Xin, WANG Meng, YU Sijiu, WANG Libin, PAN Yangyang. Research Progress on Isolation Methods and Main Reproductive Functions of Exosomes from Animal Follicular Fluid [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1480-1488.
[4]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[5]	CAO Yingying, LIANG Liang, ZENG Yirong, ZHONG Qinghua, YUAN Xiaofang, TANG Haibo, LENG Jing. Isolation,Identification and Biological Characteristics of Salmonella from Tupaia belangeri chinensis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1718-1728.
[6]	LIU Kexin, WANG Chao, ZHANG Ao, LIU Jiali, TAN Bin, ZHANG Shuqin. Isolation,Identification and Genome Sequence Analysis of Bovine Viral Diarrhea Virus LN-1 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1150-1159.
[7]	CHEN Dong, ZHANG Yanhao, ZHANG Yang, LI Sen, ZHOU Zutao, LI Zili. Isolation,Identification and Drug Sensitivity of 3 Strains of Riemerella anatipestifer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1250-1258.
[8]	WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.
[9]	XIAO Yangyang, LI Ruirui, MA Zhongchen, TANG Tian, WANG Na, CHEN Chuangfu, ZHENG Wei, WANG Yong, WANG Pengyan. Isolation,Identification and Drug Sensitivity Analysis of Mycoplasma bovis in Shihezi Region of Xinjiang [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 754-763.
[10]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[11]	SUN Yu, ZHANG Hao, LI Changyang, TAO Yanzi, WANG Qiuju. Isolation,Identification and Biological Characteristics of Antioxidant Bacillus from Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 359-367.
[12]	WEI Jin, WANG Jia, ZHAO Haoran, LIU Bo, LIU Yuanjie, YAO Wensheng, LIU Yan, MA Xin. Isolation, Identification and Antimicrobial Susceptibility of Mycoplasma gallisepticum from Different Areas [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 368-376.
[13]	YANG Fumei, YANG Linfu, ZENG Cheng, YANG Zhi, WANG Kang, DUAN Gang, DAI Feiyan. Isolation,Identification and Biological Characterization Analysis of a Strain of Carnobacterium maltaromaticum from Barking Deer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 398-407.
[14]	ZHANG Wei, WU Hua, YIN Yajie, LI Songli, HOU Shaohua. Isolation and Identification of Novel Duck Reovirus and Its σC Gene Sequence Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3520-3529.
[15]	XI Jing, YI Jihai, WANG Yueli, XU Zhenyu, LI Peidong, ZHANG Jiangwei, CHEN Chuangfu. Isolation, Identification and Genotype Analysis of Group A Bovine Rotavirus G10 Strain XJ-2022 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3530-3538.