天津地区2型猪链球菌的分离鉴定及耐药性分析
|
摘要
为了解天津地区2型猪链球菌的流行及耐药情况,本研究收集该地区部分规模化养猪场疑似猪链球菌病病料317份,通过病原分离培养、染色特性观察、生化试验、PCR扩增等方法进行鉴定,并对分离菌进行致病性及药敏试验。结果从样品中分离鉴定出11株2型猪链球菌。对小鼠的致病力试验结果显示,应用0.5×10~8 CFU/mL的细菌攻击小鼠,11株2型猪链球菌分离株中有6株具有致死性,其中1株致死率较强,5株致死率较弱。药敏试验结果表明,11株分离菌对9种临床常见药物均表现出很强的耐药性,其中对阿米卡星、四环素和强力霉素耐药性最强,耐药率达到100.00%;且所有分离株均呈多重耐药,其中4个分离株为9重耐药,占36.36%(4/11)。本试验结果表明,天津地区存在2型猪链球菌感染情况,且对多种抗菌药均产生了耐药性,应引起养殖户的高度重视,在防制猪链球菌病时,应有针对性地合理、科学、规范用药和交替用药。
In order to investigate the prevalence and drug resistance of Streptococcus suis serotype2(SS2)in Tianjin,317 samples of suspected Streptococcus suis disease were collected from largescale pig farms in this area.11 SS2 strains from the samples were identified by cultural character,dyeing property analysis,biochemical test and PCR amplification.The pathogenicity and drug susceptibility test of the isolates were also carried out.The pathogenicity test result showed that 6 strains were lethal to mice in the 11 SS2 isolate strains,including 1 strain with high lethality and 5 strains with low lethality.Drug sensitivity test result indicated the 11 isolates were highly resistant to clinical drugs,the resistance rate was up to 100.00%to amikacin,tetracycline and doxycycline.All of the 11 isolates were multidrug resistant,and 4 SS2 isolates were resistant to 9 drugs among them,which was accounting for 36.36%(4/11).The results showed that SS2 occurred in Tianjin area and the pathogenic bacteria were resistant to many antibacterials.Farmers shouldpaid close attention to this,rational,scientific,standardized medication and alternative medication when prevented the Streptococcus suis disease.
In order to investigate the prevalence and drug resistance of Streptococcus suis serotype2(SS2)in Tianjin,317 samples of suspected Streptococcus suis disease were collected from largescale pig farms in this area.11 SS2 strains from the samples were identified by cultural character,dyeing property analysis,biochemical test and PCR amplification.The pathogenicity and drug susceptibility test of the isolates were also carried out.The pathogenicity test result showed that 6 strains were lethal to mice in the 11 SS2 isolate strains,including 1 strain with high lethality and 5 strains with low lethality.Drug sensitivity test result indicated the 11 isolates were highly resistant to clinical drugs,the resistance rate was up to 100.00%to amikacin,tetracycline and doxycycline.All of the 11 isolates were multidrug resistant,and 4 SS2 isolates were resistant to 9 drugs among them,which was accounting for 36.36%(4/11).The results showed that SS2 occurred in Tianjin area and the pathogenic bacteria were resistant to many antibacterials.Farmers shouldpaid close attention to this,rational,scientific,standardized medication and alternative medication when prevented the Streptococcus suis disease.
引文
[1]刘琪,王娟,周月如,等.广东地区健康猪群和发病猪群猪链球菌流行病学调查分析[J].中国畜牧兽医,2017,44(6):1825-1831.LIU Q,WANG J,ZHOU Y R,et al.The Streptococcus suis epidemiological analysis of healthy and infected swine in Guangdong province[J].China Animal Husbandry&Veterinary Medicine,2017,44(6):1825-1831.(in Chinese)
[2]付梦玉,林丽萍,郭道出,等.一株临床2型猪链球菌的分离鉴定及生物学特性分析[J].中国畜牧兽医,2015,42(7):1897-1904.FU M Y,LIN L P,GUO D C,et al.Isolation,identification and biological characteristics analysis of a strain of clinical Streptococcus suis serotype 2[J].China Animal Husbandry&Veterinary Medicine,2015,42(7):1897-1904.(in Chinese)
[3]GOTTSCHALK M,SEGURA M.The pathogenesis of the meningitis caused be SS:Unresolved question[J].Veterinary Microbiology,2000,76(3):259-272.
[4]梁金华.猪链球菌2型广东株分离鉴定与毒力基因检测[J].畜牧与兽医,2015,47(5):1453-1457.LIANG J H.Isolation,identification and the detection of virulence genes of Streptococcus suis serotype 2in Guangdong[J].Animal Husbandry&Veterinary Medicine,2015,47(5):1453-1457.(in Chinese)
[5]魏静元,刘丽丽,张巧灵.猪Ⅱ型链球菌分离鉴定与药敏试验[J].中国农学通报,2012,28(14):105-109.WEI J Y,LIU L L,ZHANG Q L,et al.Isolation and identification of Streptococcus suis serotypes 2 and susceptibility test[J].Chinese Agricultural Science Bulletin,2012,28(14):105-109.(in Chinese)
[6]SMITH H E,VEENBERGEN V,VAN DER VELDE J,et al.The cps genes of Streptococcus suis serotypes 1,2,and 9:Development of rapid serotype-specific PCR assays[J].Journal of Clinical Microbiology,1999,37(10):3146-3152.
[7]CLSI.VET01-A4.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals,approved standard,fourth edition[S].2013.
[8]李建达,于江,张玉玉,等.猪链球菌2型实时荧光定量PCR检测方法的建立与应用[J].中国畜牧兽医,2016,43(12):3107-3113.LI J D,YU J,ZHANG Y Y,et al.Establishment and application of quantitative Real-time PCR method to detect Streptococcus suis serotype 2[J].China Animal Husbandry&Veterinary Medicine,2016,43(12):3017-3113.(in Chinese)
[9]HAN D U,CHOI C,HAM H J,et al.Prevalence,capsular type and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs in Korea[J].Canadian Journal of Veterinary Research,2001,65(3):151-155.
[10]MARIE J,MORVAN H,BERTHELOT H F,et al.Antimicrobial susceptibility of Streptococcus suis isolated from swine in France and from humans in different countries between 1996and 2000[J].Journal of Antimicrobial Chemotherapy,2002,50(2):201-209.
[11]VELA A I,MORENO M A,CEBOLLA J A,et al.Antimicrobial susceptibility of clinical strains of Streptococcus suis isolated from pigs in Spain[J].Veterinary Microbiology,2005,105(2):143-147.
[12]MAMATA G,MIGMA D T,DONG C M,et al.Molecular basis of resistance to selected antimicrobial agents in the emerging zoonotic pathogen Streptococcus suis[J].Journal of Clinical Microbiology,2015,53(7):2332-2336.
[13]苏良,欧新华,杨柳青,等.猪链球菌长沙分离株的鉴定与16SrRNA系统进化分析[J].中国人兽共患病学报,2013,29(6):547-550.SU L,OU X H,YANG L Q,et al.Identification and16SrRNA phylogenetic analysis of 2 Streptococcus suis stains isolated in Changsha city[J].Chinese Journal of Zoonoses,2013,29(6):547-550.(in Chinese)
[14]焦安心,许大文,赵洪武,等.猪2型链球菌安徽分离株的毒力基因、溶血性及耐药性分析[J].中国人兽共患病学报,2014,30(2):1181-1186.JIAO A X,XU D W,ZHAO H W,et al.Virulence genes,hemolytic and antibiotic resistance in Streptococcus suis serotype 2isolated from Anhui province,China[J].Chinese Journal of Zoonoses,2014,30(2):1181-1186.(in Chinese)
[15]李彩英,岳修为,侯怡轩,等.上海地区31株2型猪链球菌的分离鉴定及其耐药性分析[J].中国动物传染病学报,2011,19(6):54-59.LI C Y,YUE X W,HOU Y X,et al.Isolation of Streptococcus suis type 2in Shanghai area and testing of drug resistance[J].Chinese Journal of Animal Infectious Diseases,2011,19(6):54-59.(in Chinese)
[16]王娟,贾爱卿,张玮,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 Science,2017,37(6):1092-1097.(in Chinese)
[17]张伟丽,孔海深,杨青,等.Mohnarin 2010年度报告:华北地区细菌耐药监测[J].中国医院感染学杂志,2011,21(23):4909-4914.ZHANG W L,KONG H S,YANG Q,et al.Mohnarin report of 2010:Bacterial distribution and resistance in North China[J].Chinese Journal of Nosocomiology,2011,21(23):4909-4914.(in Chinese)
[18]董晓静,吕秀芝,柳艳萍,等.天津市2013年人感染猪链球菌病的首次发现和证实[J].中华流行病学杂志,2013,34(11):1154-1155.DONG X J,LV X Z,LIU Y P,et al.The report on the first case of swine Streptococcus suis type 2in Tianjin[J].Chinese Journal of Epidemiology,2013,34(11):1154-1155.(in Chinese)
[19]李哲,杨久强,葛慎峰,等.天津地区猪链球菌病病原分离[J].当代畜牧,2014,8:74-75.LI Z,YANG J Q,GE S F,et al.The pathogen separation of Streptococcus suis disease in Tianjin area[J].Contemporary Animal Husbandry,2014,8:74-75.(in Chinese)
[20]徐引弟,王志方,朱文豪,等.河南省规模化猪场猪链球菌2型的分离鉴定和药敏试验[J].中国畜牧兽医,2010,37(1):155-157.XU Y D,WANG Z F,ZHU W H,et al.Isolation,identification and drug susceptibility of Streptococcus suis2from large-scale pig farms in Henan province[J].China Animal Husbandry&Veterinary Medicine,2010,37(1):155-157.(in Chinese)
[21]彭凌,杨旭夫,朱必凤,等.猪链球菌广东株的分离鉴定及药敏试验[J].动物医学进展,2015,36(2):130-133.PENG L,YANG X F,ZHU B F,et al.Isolation,identification and drug sensitivity test of isolates of Streptococcus suis in Guangdong province[J].Progress in Veterinary Medicine,2015,36(2):130-133.(in Chinese)
[22]ZHANG C,ZHANG Z,SONG L,et al.Antimicrobial resistance profile and genotypic characteristics of Streptococcus suis capsular type 2isolated from clinical carrier sows and diseased pigs in China[J].Biomed Research International,2015,2015:284303.
[23]谭仕旦,刘国平.湖北地区猪链球菌2型的鉴定及生物学特性研究[J].湖北农业科学,2017,56(14):2722-2724.TAN S D,LIU G P.Identification and biological characteristic of Streptococcus suis type 2in Hubei[J].Hubei Agricultural Sciences,2017,56(14):2722-2724.(in Chinese)
[2]付梦玉,林丽萍,郭道出,等.一株临床2型猪链球菌的分离鉴定及生物学特性分析[J].中国畜牧兽医,2015,42(7):1897-1904.FU M Y,LIN L P,GUO D C,et al.Isolation,identification and biological characteristics analysis of a strain of clinical Streptococcus suis serotype 2[J].China Animal Husbandry&Veterinary Medicine,2015,42(7):1897-1904.(in Chinese)
[3]GOTTSCHALK M,SEGURA M.The pathogenesis of the meningitis caused be SS:Unresolved question[J].Veterinary Microbiology,2000,76(3):259-272.
[4]梁金华.猪链球菌2型广东株分离鉴定与毒力基因检测[J].畜牧与兽医,2015,47(5):1453-1457.LIANG J H.Isolation,identification and the detection of virulence genes of Streptococcus suis serotype 2in Guangdong[J].Animal Husbandry&Veterinary Medicine,2015,47(5):1453-1457.(in Chinese)
[5]魏静元,刘丽丽,张巧灵.猪Ⅱ型链球菌分离鉴定与药敏试验[J].中国农学通报,2012,28(14):105-109.WEI J Y,LIU L L,ZHANG Q L,et al.Isolation and identification of Streptococcus suis serotypes 2 and susceptibility test[J].Chinese Agricultural Science Bulletin,2012,28(14):105-109.(in Chinese)
[6]SMITH H E,VEENBERGEN V,VAN DER VELDE J,et al.The cps genes of Streptococcus suis serotypes 1,2,and 9:Development of rapid serotype-specific PCR assays[J].Journal of Clinical Microbiology,1999,37(10):3146-3152.
[7]CLSI.VET01-A4.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals,approved standard,fourth edition[S].2013.
[8]李建达,于江,张玉玉,等.猪链球菌2型实时荧光定量PCR检测方法的建立与应用[J].中国畜牧兽医,2016,43(12):3107-3113.LI J D,YU J,ZHANG Y Y,et al.Establishment and application of quantitative Real-time PCR method to detect Streptococcus suis serotype 2[J].China Animal Husbandry&Veterinary Medicine,2016,43(12):3017-3113.(in Chinese)
[9]HAN D U,CHOI C,HAM H J,et al.Prevalence,capsular type and antimicrobial susceptibility of Streptococcus suis isolated from slaughter pigs in Korea[J].Canadian Journal of Veterinary Research,2001,65(3):151-155.
[10]MARIE J,MORVAN H,BERTHELOT H F,et al.Antimicrobial susceptibility of Streptococcus suis isolated from swine in France and from humans in different countries between 1996and 2000[J].Journal of Antimicrobial Chemotherapy,2002,50(2):201-209.
[11]VELA A I,MORENO M A,CEBOLLA J A,et al.Antimicrobial susceptibility of clinical strains of Streptococcus suis isolated from pigs in Spain[J].Veterinary Microbiology,2005,105(2):143-147.
[12]MAMATA G,MIGMA D T,DONG C M,et al.Molecular basis of resistance to selected antimicrobial agents in the emerging zoonotic pathogen Streptococcus suis[J].Journal of Clinical Microbiology,2015,53(7):2332-2336.
[13]苏良,欧新华,杨柳青,等.猪链球菌长沙分离株的鉴定与16SrRNA系统进化分析[J].中国人兽共患病学报,2013,29(6):547-550.SU L,OU X H,YANG L Q,et al.Identification and16SrRNA phylogenetic analysis of 2 Streptococcus suis stains isolated in Changsha city[J].Chinese Journal of Zoonoses,2013,29(6):547-550.(in Chinese)
[14]焦安心,许大文,赵洪武,等.猪2型链球菌安徽分离株的毒力基因、溶血性及耐药性分析[J].中国人兽共患病学报,2014,30(2):1181-1186.JIAO A X,XU D W,ZHAO H W,et al.Virulence genes,hemolytic and antibiotic resistance in Streptococcus suis serotype 2isolated from Anhui province,China[J].Chinese Journal of Zoonoses,2014,30(2):1181-1186.(in Chinese)
[15]李彩英,岳修为,侯怡轩,等.上海地区31株2型猪链球菌的分离鉴定及其耐药性分析[J].中国动物传染病学报,2011,19(6):54-59.LI C Y,YUE X W,HOU Y X,et al.Isolation of Streptococcus suis type 2in Shanghai area and testing of drug resistance[J].Chinese Journal of Animal Infectious Diseases,2011,19(6):54-59.(in Chinese)
[16]王娟,贾爱卿,张玮,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 Science,2017,37(6):1092-1097.(in Chinese)
[17]张伟丽,孔海深,杨青,等.Mohnarin 2010年度报告:华北地区细菌耐药监测[J].中国医院感染学杂志,2011,21(23):4909-4914.ZHANG W L,KONG H S,YANG Q,et al.Mohnarin report of 2010:Bacterial distribution and resistance in North China[J].Chinese Journal of Nosocomiology,2011,21(23):4909-4914.(in Chinese)
[18]董晓静,吕秀芝,柳艳萍,等.天津市2013年人感染猪链球菌病的首次发现和证实[J].中华流行病学杂志,2013,34(11):1154-1155.DONG X J,LV X Z,LIU Y P,et al.The report on the first case of swine Streptococcus suis type 2in Tianjin[J].Chinese Journal of Epidemiology,2013,34(11):1154-1155.(in Chinese)
[19]李哲,杨久强,葛慎峰,等.天津地区猪链球菌病病原分离[J].当代畜牧,2014,8:74-75.LI Z,YANG J Q,GE S F,et al.The pathogen separation of Streptococcus suis disease in Tianjin area[J].Contemporary Animal Husbandry,2014,8:74-75.(in Chinese)
[20]徐引弟,王志方,朱文豪,等.河南省规模化猪场猪链球菌2型的分离鉴定和药敏试验[J].中国畜牧兽医,2010,37(1):155-157.XU Y D,WANG Z F,ZHU W H,et al.Isolation,identification and drug susceptibility of Streptococcus suis2from large-scale pig farms in Henan province[J].China Animal Husbandry&Veterinary Medicine,2010,37(1):155-157.(in Chinese)
[21]彭凌,杨旭夫,朱必凤,等.猪链球菌广东株的分离鉴定及药敏试验[J].动物医学进展,2015,36(2):130-133.PENG L,YANG X F,ZHU B F,et al.Isolation,identification and drug sensitivity test of isolates of Streptococcus suis in Guangdong province[J].Progress in Veterinary Medicine,2015,36(2):130-133.(in Chinese)
[22]ZHANG C,ZHANG Z,SONG L,et al.Antimicrobial resistance profile and genotypic characteristics of Streptococcus suis capsular type 2isolated from clinical carrier sows and diseased pigs in China[J].Biomed Research International,2015,2015:284303.
[23]谭仕旦,刘国平.湖北地区猪链球菌2型的鉴定及生物学特性研究[J].湖北农业科学,2017,56(14):2722-2724.TAN S D,LIU G P.Identification and biological characteristic of Streptococcus suis type 2in Hubei[J].Hubei Agricultural Sciences,2017,56(14):2722-2724.(in Chinese)