鸡滑液囊支原体贵州株的分离鉴定及VlhA基因序列分析

doi:10.16431/j.cnki.1671-7236.2022.04.034

Abstract

Abstract: 【Objective】 The aim of this study was to obtain Mycoplasma synoviae(MS) strains in Guizhou,to further develop its etiology and immunology related research.【Method】 A total of 107 samples of pharyngeal swabs,swollen joints and claw pads and swollen tissue exudate suspected to be infect with MS were collected from a broiler farm in Zhijin county,Guizhou province and detected by PCR.The samples tested positive by PCR were used to isolate and culture MS,Wright staining,biochemical test and sequence analysis of the VlhA gene were performed.【Result】 20 positive samples were tested by PCR,only 1 strain of MS was isolated from the positive samples of joint and claw pad,and the colony formed "egg-shaped" on solid medium.The isolated strain was small blue spherical stained by Wright’s in the microscope.In biochemical tests,the isolates could decompose glucose and maltose,but not arginine and urea,which was consistent with the biochemical results of MS standard strains.The target fragment of 821 bp was amplified by PCR.Nucleotide similarity analysis of the VlhA gene of the isolated strain indicated that the strain was MS,and there were regional differences in similarity with other endemic strains in GenBank.The similarity with the epidemic strains in Anhui,Chongqing,Fujian,Guangdong,Guangxi,Hubei,Hunan,Jiangsu and Yunnan was 90.3% to 99.7%.The most similar strain was MS epidemic strain in Hunan,which reached 99.7%.But the similarity with foreign reference strains was low.The phylogenetic analysis of VlhA gene showed that this strain was closely related to the domestic reference strain,but far from the foreign.【Conclusion】 In this study,a Guizhou endemic MS strain was successfully isolated and identified from chickens causing swelling of joints and foot pads with some variation.This study provided a basis for the molecular and biological characteristics of MS in Guizhou and the diagnosis and control of this disease.

Key words: Mycoplasma synoviae; isolation; identification; sequence analysis

CLC Number:

S852.62

SONG Chun, WANG Bolin, LI Mei, WEN Ming, WANG Kaigong, CHEN Changxiu, CHENG Zhentao. Isolation and Identification of Mycoplasma synoviae Guizhou Strain and Sequence Analysis of VlhA Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1516-1523.

References

[1] PASCUCCI S,MAESTRINI N,GOVONI S,et al.Mycoplasma synoviae in the guinea-fowl[J].Avian Pathology,1976,5(4):291-297.
[2] KORDAFSHARI S,MARENDA M S,O’ROURKE D,et al.Mutation of oppF gene in the Mycoplasma synoviae MS-H vaccine strain and its implication for differential serological responses to vaccination versus field challenge[J].Veterinary Microbiology,2019,231:48-55.
[3] ZOFIA L,SAWOMIR P,OLIMPIA K,et al.Spectral technique for detection of changes in eggshells caused by Mycoplasma synoviae[J].Poultry Science,2019,98(9):3481-3487.
[4] 曹中赞,栾新红,刘胜旺,等.禽滑液囊支原体研究进展[J].动物医学进展,2012,33(11):113-117. CAO Z Z,LUAN X H,LIU S W,et al.Advance in avian Mycoplasma synoviae[J].Progress in Veterinary Medicine,2012,33(11):113-117.(in Chinese)
[5] 鲜思美,汪德生,任晓江,等.鸡滑液囊支原体病继发大肠杆菌感染的诊断[J].畜牧与兽医,2013,45(7):80-82. XIAN S M,WANG D S,REN X J,et al.Diagnosis of Escherichia coli co-infection to Mycoplasma synoviae[J].Animal Husbandry and Veterinary Medicine,2013,45(7):80-82.(in Chinese)
[6] AMER M M,MEKKY H M,FEDAWY H S.Molecular identification of Mycoplasma synoviae from breeder chicken flock showing arthritis in Egypt[J].Veterinary World,2019,12(4):535-541.
[7] 石晓磊.鸡滑液囊支原体的分离鉴定及其活疫苗免疫效果的评价[D].杨凌:西北农林科技大学,2018. SHI X L.Isolation and identification of Mycoplasma synoviae and evaluation of the effect of live-vaccine[D].Yangling:Northwest A&F University,2018.(in Chinese)
[8] 田亚琴,文玉康,王豪举,等.鸡滑液囊支原体疫苗研究进展[J].中国家禽,2020,42(9):96-102. TIAN Y Q,WEN Y K,WANG H J,et al.Research progress in the development of Mycoplasma synoviae vaccine[J].China Poultry,2020,42(9):96-102.(in Chinese)
[9] 招丽婵,覃健萍,王占新,等.鸡滑液囊支原体的流行调查及不同地区分离株药物敏感性分析[J].中国兽医杂志,2019,55(6):9-13. ZHAO L C,QIN J P,WANG Z X,et al.Epidemic investigation and drug sensitivity analysis of Mycoplasma synoviae from different geographical locations[J].Chinese Journal of Veterinary Medicine,2019,55(6):9-13.(in Chinese)
[10] 任峰,谭磊,包世俊,等.鸡滑液支原体乳酸脱氢酶的克隆表达及其表达产物活性研究[J].中国动物传染病学报,2014,22(2):52-57. REN F,TAN L,BAO S J,et al.Cloning and expression of lactate dehydrogenase gene of Mycoplasma synoviae and determination of enzymatic activity of the recombinant protein[J].Chinese Journal of Animal Infectious Diseases,2014,22(2):52-57.(in Chinese)
[11] 丁美娟,尹秀凤,张海涛,等.江苏部分地区鸡滑液囊支原体感染的血清学调查[J].中国家禽,2015,37(11):73-74. DING M J,YIN X F,ZHANG H T,et al.Serological investigation of Mycoplasma synoviae infection in Jiangsu[J].China Poultry,2015,37(11):73-74.(in Chinese)
[12] 陈秀红,郭亚男,司朵朵,等.宁夏地区鸡滑液囊支原体病流行病学调查与分析[J].中国预防兽医学报,2020,42(3):234-238. CHEN X H,GUO Y N,SI D D,et al.Epidemiological investigation and analysis of Mycoplasma synoviae in infection in chickens in Ningxia region[J].Chinese Journal of Preventive Veterinary Medicine,2020,42(3):234-238.(in Chinese)
[13] 王艳丰,张丁华,朱金凤.鸡滑液囊支原体病流行现状及防控技术研究进展[J].中国畜牧兽医,2021,48(8):3038-3049. WANG Y F,ZHANG D H,ZHU J F.Research progress on epidemic status and prevention and control technology of Mycoplasma synoviae infection[J].China Animal Husbandry & Veterinary Medicine,2021,48(8):3038-3049.(in Chinese)
[14] 丁美娟,尹秀凤,黄显明,等.鸡滑液囊支原体病研究进展[J].中国家禽,2013,35(13):39-41. DING M J,YIN X F,HUANG X M,et al.Research progress of Mycoplasma synoviae[J].China Poultry,2013,35(13):39-41.(in Chinese)
[15] BENCINA D,DROBNIC-VALIC M,HORVAT S,et al.Molecular basis of the length variation in the N-terminal part of Mycoplasma synoviae hemagglutinin[J].FEMS Microbiology Letters,2001,203(1):115-123.
[16] EL-GAZZAR M M,WETZEL A N,RAVIV Z.The genotyping potential of the Mycoplasma synoviae vlhA gene[J].Avian Diseases,2012,56(4):711-719.
[17] 李凡,张焕容,陈云霞.川西地区鸡滑液囊支原体分离株的部分生物学特性及其VlhA基因遗传进化分析[J].中国预防兽医学报,2020,42(2):122-127. LI F,ZHANG H R,CHEN Y X.Partial biological characteristics of Mycoplasma synoviae isolates from Western Sichuan analysis of it VlhA gene[J].Chinese Journal of Preventive Veterinary Medicine,2020,42(2):122-127.(in Chinese)
[18] 田浪,温贵兰,程振涛,等.鸡滑液囊支原体的RT-PCR检测及基因序列分析[J].贵州畜牧兽医,2019,43(1):45-47. TIAN L,WEN G L,CHENG Z T,et al.RT-PCR detection and gene sequence analysis of Mycoplasma synoviae[J].Guizhou Journal of Animal Husbandry & Veterinary Medicine,2019,43(1):45-47.(in Chinese)
[19] 丁美娟.鸡滑液囊支原体的分离鉴定及部分生物学特性研究[D].南京:南京农业大学,2013. DING M J.Isolation and identification of Mycoplasma synoviae and its partial biological characteristics[D].Nanjing:Nanjing Agricultural University,2013.(in Chinese)
[20] ZHU L,SHAHID M A,MARKHAM J,et al.Genome analysis of Mycoplasma synoviae strain MS-H,the most common M.synoviae strain with a worldwide distribution[J].BMC Genomics,2018,19(1):117.
[21] 徐引弟,焦文强,王治方,等.鸡滑液囊支原体河南株的分离鉴定与生物学特性研究[J].河南农业科学,2021,50(7):168-173. XU Y D,JIAO W Q,WANG Z F,et al.Isolation,identification and biological characteristics of Mycoplasma synoviae from Henan strain[J].Henan Agricultural Sciences,2021,50(7):168-173.(in Chinese)
[22] SUN S,LIN X,LIU J,et al.Phylogenetic and pathogenic analysis of Mycoplasma synoviae isolated from native chicken breeds in China[J].Poultry Sciences,2017,96(7):2057-2063.
[23] 丁美娟,张小飞,尹秀凤,等.鸡滑液囊支原体的分离和鉴定研究[J].中国家禽,2011,33(24):27-29. DING M J,ZHANG X F,YIN X F,et al.Isolation and identification of Mycoplasma synoviae from broiler breeders[J].China Poultry,2011,33(24):27-29.(in Chinese)
[24] LIMPA K,SASIPREEYAJAN J,PAKPINYO S.Characterization of Thai Mycoplasma synoviae isolates by sequence analysis of partial vlhA gene[J].Avian Diseases,2016,60(4):810-816.
[25] 冯霞,张富强,徐福阳,等.人芽囊原虫感染动物模型构建研究进展[J].中国血吸虫病防治杂志,2021,33(1):102-106. FENG X,ZHANG F Q,XU F Y,et al.Progress of researches on animal models of Blastocystis hominis infections[J].Chinese Journal of Schistosomiasis Control,2021,33(1):102-106.(in Chinese)
[26] 侯博,王晨燕,邵国青.滑液囊支原体不同感染途径的致病力比较[J].微生物学通报,2021,48(8):2704-2713. HOU B,WANG C Y,SHAO G Q.Comparison of pathogenicity to Mycoplasma synoviae with different injection routes[J].Microbiology China,2021,48(8):2704-2713.(in Chinese)

Isolation and Identification of Mycoplasma synoviae Guizhou Strain and Sequence Analysis of VlhA Gene

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