致狐狸肺炎大肠杆菌血清鉴定与毒力基因检测
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摘要
为了确定狐狸肺炎源致病性大肠杆菌(Escherichia coli,E.coli)流行血清型与毒力基因的携带情况并分析其相关性,2016—2017年采集河北省部分地区患肺炎死亡的狐狸的肺脏、心血等组织142份中分离到了105株E.coli,且RT-PCR检测犬瘟热病毒为阴性,人工感染小鼠致病性试验结果显示105株分离菌株中77株为致病性E.coli。采用玻片凝集试验和PCR方法分别检测77株致病性E.coli血清型和毒力基因,结果显示:77株致病性E.coli有9种血清型,其中以O1、O8、O78、O12为流行的优势致病血清型,77株致病性E.coli携带14种不同毒力基因,以irp2、fyuA、papC、fimC、iucD~a、colv、hlyF、ompT 8种毒力基因检出率较高,在32.5%～98.7%之间,其余毒力基因检出率较低。狐狸肺炎源致病性E.coli的优势血清型携带的毒力基因较多,至少携带9种毒力基因,流行的优势血清型与毒力基因之间具有一定的相关性。狐狸肺炎源E.coli流行血清型代表株TS(O1)1、QH(O8)2、QH(O78)3、TS(O12)4均对狐狸很强的致病性。本试验为狐狸肺炎源致病性E.coli防控提供了参考依据。
To determine the prevalence of serotype and virulence genes in the E.coli of fox pneumonia and to analyze its correlation,105 strains of E.coli were isolated from 142 parts of the lung,heart and blood of the fox which died of pneumonia in some areas of Hebei province in 2016-2017.The detection of canine distemper virus was negative by RT-PCR.The results of pathogenicity test of artificial infected mice showed that the 77 isolates were pathogenic E.coli.77 pathogenicity E.coli serotypes and virulence genes were detected by slide agglutination test and PCR method,respectively.The results showed that 77 strains of pathogenic E.coli blong to 9 serotypes,among which O1,O8,O78,O12 were as predominant serotypes,77 strains of pathogenic E.coli carried 14 different virulence genes,The positive rate of irp2,fyuA,papC,fimC,iucDa,colv,hlyF,ompT 8 of virulence genes was 32.5%-98.7%,the other virulence gene were low.The dominant serotype of fox pneumonia pathogenic E.coli has many virulence genes,carrying at least 9 virulence genes.There was a certain correlation between the dominant serotypes and virulence genes.The results showed that the fox pneumonia source E.coli epidemic serotype TS(O1) 1,QH(O8) 2,QH(O78) 3,and TS(O12) 4 were all highly pathogenic to foxes.This study provides a reference for the prevention and control of the pathogenic E.coli of fox pneumonia.
To determine the prevalence of serotype and virulence genes in the E.coli of fox pneumonia and to analyze its correlation,105 strains of E.coli were isolated from 142 parts of the lung,heart and blood of the fox which died of pneumonia in some areas of Hebei province in 2016-2017.The detection of canine distemper virus was negative by RT-PCR.The results of pathogenicity test of artificial infected mice showed that the 77 isolates were pathogenic E.coli.77 pathogenicity E.coli serotypes and virulence genes were detected by slide agglutination test and PCR method,respectively.The results showed that 77 strains of pathogenic E.coli blong to 9 serotypes,among which O1,O8,O78,O12 were as predominant serotypes,77 strains of pathogenic E.coli carried 14 different virulence genes,The positive rate of irp2,fyuA,papC,fimC,iucDa,colv,hlyF,ompT 8 of virulence genes was 32.5%-98.7%,the other virulence gene were low.The dominant serotype of fox pneumonia pathogenic E.coli has many virulence genes,carrying at least 9 virulence genes.There was a certain correlation between the dominant serotypes and virulence genes.The results showed that the fox pneumonia source E.coli epidemic serotype TS(O1) 1,QH(O8) 2,QH(O78) 3,and TS(O12) 4 were all highly pathogenic to foxes.This study provides a reference for the prevention and control of the pathogenic E.coli of fox pneumonia.
引文
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[17] 夏琦琦,温珊珊,马斯琪,等.水貂源致病性大肠杆菌生物被膜形成能力、耐药性及毒力岛基因检测与分析[J].中国预防兽医学报,2017,39(5):361-365.
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[20] 张美君,廖晓萍,王秀梅,等.宠物源大肠杆菌的血清型和毒力基因及耐药性调查[J].中国预防兽医学报,2011,33(8):601-605.
[2] 张召兴,李蕴玉,张艳英,等.秦皇岛貉源E.coli的分离鉴定与耐药性分析[J].黑龙江畜牧兽医,2016,(6):175-177.
[3] 马增军,芮萍,逯春香,等.猪源肠外致病性大肠杆菌的分离与鉴定[J].中国人畜共患病学报,2015,31(2):130-134.
[4] 郭蕊,项方,张艳英,等.狐狸大肠杆菌病的诊断 [J].Agricultural Science & Technology,2016,17(1):161-163+227.
[5] 张志强,吴同垒,李永慧,等.狐肺炎大肠杆菌HtrA蛋白原核表达及多抗血清制备[J].中国兽医学报,2017,37(7):1255-1260.
[6] 杨斯琴,敖日格乐,王纯洁,等.内蒙古呼和浩特地区牛源致病性大肠杆菌的血清型、毒力基因检测及耐药性分析[J].中国预防兽医学报,2015,37(10):761-764.
[7] 张召兴,李蕴玉,吴同垒,等.邢台地区致鸡卵黄性腹膜炎大肠杆菌血清型鉴定、毒力基因检测及耐药性分析[J].中国预防兽医学报,2017,39(3):191-195.
[8] LAU M E,DANKA E S,TIEMANN K M.et al.Bacterial lysis liberates the neutrophil migration suppressor YbcL from the periplasm of uropathogenic Escherichia coli[J].Infect Immun,2014,82(12):4921–4930.
[9] 孙彦刚,赵建军,史宁,等.狐源犬瘟热病毒的分离鉴定及H 基因遗传变异分析[J].中国预防兽医学报,2016,38(3):206-210.
[10] 张召兴,贾青辉,李蕴玉,等.从死胎流产的母貉体内检出1株犬链球菌[J].中国兽医学报,2016,36(6):957-960.
[11] 罗燕,王朋,赵洪明,等.林麝肺源致病性大肠杆菌分离鉴定及毒力基因PCR检测[J].中国预防兽医学报,2012,34(8):615-618.
[12] 芮萍,马增军,杨彩然,等.狐貉貂源致病性大肠杆菌的分离与鉴定[J].中国兽医科学,2015,45(5):517-521.
[13] 周鑫,罗燕,程建国,等.林麝肺源致病性大肠杆菌O因子血清型鉴定及部分耐药基因的PCR检测[J].四川动物,2014,33(5):715-720.
[14] JAKOBSEN L,SPANGHOLM D J,PEDERSEN K,et al.Broiler chickens,broiler chicken meat,pigs and pork as sources of ExPEC related virulence genes and resistance in Escherichia coli isolates from community-dwelling humans and UTI patients[J].In J Food Microbiol,2014,142(1/2):264-272.
[15] JOHNSON T J,WANNEMUEHLER Y,DOETKOTT C,et al.Identification of minimal predictors of avian pathogenic Escherichia coli virulence for use as a rapid diagnostic tool[J].J Clin Microbiol,2008,46(12):3987-3996.
[16] HAENNI M,CHATRE P,METAYER V,et al.Comparative prevalence and characterization of ESBL-producing Enterobacteriaceae in dominant versus subdominant enteric flora in veal calves at slaughterhouse,France [J].Vet Microbiol,2014,171(3/4):321-327.
[17] 夏琦琦,温珊珊,马斯琪,等.水貂源致病性大肠杆菌生物被膜形成能力、耐药性及毒力岛基因检测与分析[J].中国预防兽医学报,2017,39(5):361-365.
[18] Asai T,Masani K,Sato C,et al.Phylogenetic groups and cephalosporin resistance of Escherichia coli from diseased food-producing animals in Japan[J].Acta Vet Scand,2011,53(1):52.
[19] 张召兴,李蕴玉,贾青辉,等.鸡致病性大肠杆菌病的研究进展[J].河北科技师范学院学报,2017,31(2):68-72+76.
[20] 张美君,廖晓萍,王秀梅,等.宠物源大肠杆菌的血清型和毒力基因及耐药性调查[J].中国预防兽医学报,2011,33(8):601-605.