广西部分地区猪链球菌血清型及其毒力因子流行性调查分析
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摘要
为探究广西部分地区猪链球菌(Streptococcus suis,SS)流行菌株主要血清型及毒力因子分布情况。本试验于2018年1月至2018年6月对从多个猪场采集到的116份疑似链球菌感染的组织病料(脑、肺脏、淋巴结等)进行病原菌检测,采用细菌分离鉴定、形态学观察及PCR扩增等方法对病原菌及其血清型和部分毒力因子进行鉴定。结果显示,116份样品共分离到链球菌32株,阳性率为27.59%(32/116),其血清型主要以SS2和SS9为主,分离率分别为40.63%(13/32)和43.75%(14/32),其他血清型为15.63%(5/32);毒力因子检测结果表明:SLY、MRP、EPF以及SBP2′因子的检出率分别为:81.25%(26/32)、59.38%(19/32)、50.00%(16/32)和71.88%(23/32)。32株链球菌中以SLY~+MRP~+EPF~+SBP2′~+(13株)、SLY~+MRP~-EPF~-SBP2′~-(5株)、SLY~+MRP~+EPF~-SBP2′~+(5株)为主要的毒力基因型,其中SS2均能检测出3个或3个以上的毒力因子。玉林、柳州市主要分布SS9型,南宁、百色市主要分布SS2型;广西不同地区毒力因子的分布情况存在差异且不同血清型或同一血清型的链球菌毒力因子的分布情况也各不相同,其中SS2携带的毒力因子检出率明显高于其他血清型。该研究可为今后猪链球菌疫苗及致病机理研究提供理论依据,为广西地区猪场链球菌血清型疫苗选择提供指导。
To investigate the distribution of major serotypes and virulence factors of Streptococcus suis(SS) strains in parts regions of Guangxi,116 samples of suspected streptococcal infections(brain,lung,lymph nodes,etc.) collected from different swine farms from January to June in 2018 were examined in this study.The serotypes and some virulence genes were identified by bacterial isolation,morphological observation,PCR amplification.The positive rate of Streptococcus suis in 116 samples was 27.59%(32/116).The main serotypes of positive samples were SS2(40.63%,13/32) and SS9(43.75%,14/32),and other serotypes accounted for 15.63%(5/32).The test results of virulence factors demonstrated that the detection rates of SLY,MRP,EPF and SBP2′ factors were 81.25%(26/32),59.38%(19/32),50.00%(16/32) and 71.88%(23/32),respectively.Among the 32 strains of Streptococcus suis,SLY~+MRP~+EPF~+SBP2′~+(13 strains),SLY~+MRP~-EPF~-SBP2′~-(5 strains) and SLY~+MRP~+EPF~-SBP2′~+(5 strains) were the main virulence genotypes.And SS2 could be detected three or more virulence genes.SS9 was the prevalent type in Yulin and Liuzhou.SS2 was the prevalent type in Nanning and Baise.It was discovered that there were differences in the distribution of virulence factors in the four regions of Guangxi.The distribution of streptococcal virulence factors in different serotypes or the same serotype was different,and the detection rate of virulence gene of SS2 was higher than other serotypes.This study could provide a theoretical basis for the future research of Streptococcus suis vaccine and pathogenesis,and provide guidance for the selection of serotypes of Streptococcus suis vaccine in Guangxi.
To investigate the distribution of major serotypes and virulence factors of Streptococcus suis(SS) strains in parts regions of Guangxi,116 samples of suspected streptococcal infections(brain,lung,lymph nodes,etc.) collected from different swine farms from January to June in 2018 were examined in this study.The serotypes and some virulence genes were identified by bacterial isolation,morphological observation,PCR amplification.The positive rate of Streptococcus suis in 116 samples was 27.59%(32/116).The main serotypes of positive samples were SS2(40.63%,13/32) and SS9(43.75%,14/32),and other serotypes accounted for 15.63%(5/32).The test results of virulence factors demonstrated that the detection rates of SLY,MRP,EPF and SBP2′ factors were 81.25%(26/32),59.38%(19/32),50.00%(16/32) and 71.88%(23/32),respectively.Among the 32 strains of Streptococcus suis,SLY~+MRP~+EPF~+SBP2′~+(13 strains),SLY~+MRP~-EPF~-SBP2′~-(5 strains) and SLY~+MRP~+EPF~-SBP2′~+(5 strains) were the main virulence genotypes.And SS2 could be detected three or more virulence genes.SS9 was the prevalent type in Yulin and Liuzhou.SS2 was the prevalent type in Nanning and Baise.It was discovered that there were differences in the distribution of virulence factors in the four regions of Guangxi.The distribution of streptococcal virulence factors in different serotypes or the same serotype was different,and the detection rate of virulence gene of SS2 was higher than other serotypes.This study could provide a theoretical basis for the future research of Streptococcus suis vaccine and pathogenesis,and provide guidance for the selection of serotypes of Streptococcus suis vaccine in Guangxi.
引文
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[3] GOTTSCHALK M,HIGGINS R,JACQUES M,et al.Description of 14 new capsular types of Streptococcus suis[J].Journal of Clinical Microbiology,1989,27(12):2633-2636.
[4] 熊毅,刘棋,覃芳芸,等.猪链球菌亚种及1、2、9型四重PCR检测方法的建立[J].南方农业学报,2007,38(6):681-684. XIONG Y,LIU Q,QIN F Y,et al.Establishment of quadruple PCR for detecting subspecies and type 1,2 and 9 of swine Streptococcus suis[J].Guangxi Agricultural Sciences,2007,38(6):681-684.(in Chinese)
[5] 闫若潜,吴志明,张志凌,等.猪链球菌1、7、9型多重PCR诊断方法的建立及初步应用[J].中国畜牧兽医,2009,36(7):154-157. YAN R Q,WU Z M,ZHANG Z L,et al.Establishment and application of multiplex PCR assay for detection of Streptococcus suis serotypes 1,7 and 9[J].China Animal Husbandry & Veterinary Medicine,2009,36(7):154-157.(in Chinese)
[6] ZHANG C P,NING Y B,ZHANG Z Q,et al.Prevalence of Streptococcus suis isolated from clinically healthy sows in China[J].Agricultural Sciences in China,2009,8(5):638-642.
[7] FENG Y,ZHANG H,WU Z,et al.Streptococcus suis infection:An emerging/reemerging challenge of bacterial infectious diseases[J].Virulence,2014,5(4):477-497.
[8] GOTTSCHALK M,SEGURA M,XU J.Streptococcus suis infections in humans:The Chinese experience and the situation in North America[J].Animal Health Research Reviews,2007,8(1):29-45.
[9] OKWUMABUA O,PERSAUD J S,REDDY P G.Cloning and characterization of the gene encoding the glutamate dehydrogenase of Streptococcus suis serotype 2[J].Clinical and Diagnostic Laboratory Immunology,2001,8(2):251-257.
[10] 陈雯雯,韦显凯,闭璟珊,等.广西部分地区猪链球菌的分离鉴定及小鼠致病性研究[J].中国畜牧兽医,2017,44(3):861-867. CHEN W W,WEI X K,BI J S.et al.Isolation,identification and pathogenicity on mice of Streptococcus suis in pig in Guangxi border area[J].China Animal Husbandry & Veterinary Medicine,2017,44(3):861-867.(in Chinese)
[11] KERDSIN A,DEJSIRILERT S,AKEDA Y,et al.Fifteen Streptococcus suis serotypes identified by multiplex PCR[J].Journal of Medical Microbiology,2012,61(12):1669-1672.
[12] 谈忠鸣,鲍昌俊,潘红星,等.快速PCR检测猪链球菌2型方法的研究及应用[J].江苏预防医学,2012,23(6):9-11. TAN Z M,BAO C J,PAN H X,et al.Establishment of fast PCR assay 5 species genes for Streptococcus suis serotype 2 and its application to detect strains isolated from Jiangsu[J].Jiangsu Journal of Preventive Medicine,2012,23(6):9-11.(in Chinese)
[13] BEINEKE A,BENNECKE K,NEIS C,et al.Comparative evaluation of virulence and pathology of Streptococcus suis serotypes 2 and 9 in experimentally infected growers[J].Veterinary Microbiology,2008,128(3-4):423-430.
[14] 侯雪静.猪链球菌病流行病学及其疫苗研究现状[J].今日畜牧兽医,2018,34(7):77. HOU X J.The developing situation of vaccines and epidemiology in Streptococcus suis[J].Today Animal Husbandry and Veterinary Medicine,2018,34(7):77.(in Chinese)
[15] 吴静波,南文金,胡鸿惠.猪链球菌毒力因子致病机制研究进展[J].动物医学进展,2015,36(9):97-100. WU J B,NAN W J,HU H H.Progress on pathogenic mechanism of Streptococcus suis virulence factors[J].Progress in Veterinary Medicine,2015,36(9):97-100.(in Chinese)
[16] DONG W,MA J,ZHU Y,et al.Virulence genotyping and population analysis of Streptococcus suis serotype 2 isolates from China[J].Infection,Genetics and Evolution,2015,36:483-489.
[17] 周明瑶.2015年苏南地区健康猪群猪链球菌流行病学调查[D].南京:南京农业大学,2016. ZHOU M Y.Epidemiological survey of Streptococcus suis in healthy pigs in southern Jiangsu in 2015[D].Nanjing:Nanjing Agricultural University,2016.(in Chinese)
[18] 王福春,黄德胜,谢继杰.广西靖西市一起人感染猪链球菌病疫情调查分析[J].医学动物防制,2017,33(9):962-964. WANG F C,HUANG D S,XIE J J.Investigation and analysis of a human infection with swine streptococci in Jingxi city,Guangxi[J].Journal of Medical Pest Control,2017,33(9):962-964.(in Chinese)
[19] 李延平.猪链球菌病流行特征及防治措施[J].中国畜牧兽医文摘,2018,34(6):247. LI Y P.Epidemiological characteristics and control measures of Streptococcus suis[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2018,34(6):247.(in Chinese)
[20] 吴景文,王斌,郭双莉,等.江西省一例人感染1型猪链球菌病例的流行病学调查与分析[J].国外医学(医学地理分册),2017,38(2):164-165. WU J W,WANG B,GUO S L,et al.An epidemiological investigation of one case of human Streptococcus suis type 1 infection in Jiangxi province[J].Foreign Medical Sciences (Medical Geography),2017,38(2):164-165.(in Chinese)
[21] 马青,刘英,王月,等.贵州省死亡病例1型猪链球菌的分离及分子生物学特征分析[J].现代预防医学,2018,45(3):506-509. MA Q,LIU Y,WANG Y,et al.Identification and characterization of Streptococcus suis serotype 1 strain isolated from a death case,Guizhou[J].Modern Preventive Medicine,2018,45(3):506-509.(in Chinese)
[22] KAICHENG W,ZONGFU W,HUOCHUN Y,et al.Identification and detection of serotype-specific genes:Effective serotyping of Streptococcus suis[J].Current Clinical Microbiology Reports,2017,4(1):29-35.
[23] 马婷婷.2017年广西部分地区猪链球菌2、7、9型流行病学调查[D].南宁:广西大学,2018. MA T T.Epidemiological survey of Streptococcus suis type 2,7 and 9 in parts of Guangxi in 2017[D].Nanning:Guangxi University,2018.(in Chinese)
[24] 王娟,贾爱卿,张炜,等.广州周边地区健康猪群猪链球菌携带情况调查[J].中国兽医学报,2017,37(6):1092-1097. WANG J,JIA A Q,ZHANG W,et al.Analysis and epidemiological investigation of Streptococcus suis in Guangzhou surrounding areas[J].Chinese Journal of Veterinary Medicine,2017,37(6):1092-1097.(in Chinese)
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