保育猪舍中猪源、粪源、气源大肠杆菌的分离鉴定及16S rDNA同源性分析
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摘要
为了探究断奶仔猪大肠杆菌病的发病原因,选取山西省介休市某规模化猪场中单栋饲养量为500头并有大肠杆菌病发生的半封闭保育猪舍1栋,分别采集舍内的空气、猪的粪便和鼻腔黏液、发病猪的肺部组织样品,进行大肠杆菌的分离培养与鉴定,并对分离出的大肠杆菌进行16S rDNA基因全长测序和同源性分析。结果表明,培养鉴定后共分离出大肠杆菌21株(包括舍内空气中12株、猪的粪便中3株、猪的鼻腔黏液中3株、发病猪的肺部组织中3株),通过细菌通用引物27F和1492R,扩增到大肠杆菌16S rDNA基因全长约1 500 bp。其中,空气中的大肠杆菌与猪的粪便、鼻腔黏液、发病猪的肺部组织样品中的大肠杆菌相似度分别为98.0%～99.2%、94.9%～96.3%和98.3%～99.1%。由此可见,粪源大肠杆菌是猪舍空气环境中大肠杆菌的主要来源,且保育猪舍内大肠杆菌在猪体、粪便和舍内环境中均存在,不同源大肠杆菌16S rDNA基因之间具有较高同源性。
In order to study the pathogenesis of colibacillosis of weaned piglets,a semi-closed weaning pig house with 500 piglets and colibacillosis occurring was selected in Jiexiu city,Shanxi province.Aerosol,feces,nasal-mucus and diseased lung tissue samples were colected and Escherichia coli was isolated and identified,16 S rDNA overall length was sequenced and homology analysis of the isolated Escherichia coli was carried out.The results showed that 21 strains of Escherichia coli(including 12 strains in aerosol,3 strains in feces,3 strains in nasal-mucus,3 strains in diseased lung tissue) were isolated by microbiological identification,and 1 500 bp of 16 S rDNA overall length was amplified by primers 27 F and 1492 R.The similarity between Escherichia coli in aerosol and Escherichia coli in feces,nasal-mucus and diseased lung tissue samples of pigs were 98.0%—99.2%,94.9%—96.3% and 98.3%—99.1%,respectively.It showed that the Escherichia coli from feces was the main source of Escherichia coli in the aerosol in weaning pig house,Escherichia coli was found in piglet tissues,feces and environment,and the higher homology was possessed between different sources of Escherichia coli.
In order to study the pathogenesis of colibacillosis of weaned piglets,a semi-closed weaning pig house with 500 piglets and colibacillosis occurring was selected in Jiexiu city,Shanxi province.Aerosol,feces,nasal-mucus and diseased lung tissue samples were colected and Escherichia coli was isolated and identified,16 S rDNA overall length was sequenced and homology analysis of the isolated Escherichia coli was carried out.The results showed that 21 strains of Escherichia coli(including 12 strains in aerosol,3 strains in feces,3 strains in nasal-mucus,3 strains in diseased lung tissue) were isolated by microbiological identification,and 1 500 bp of 16 S rDNA overall length was amplified by primers 27 F and 1492 R.The similarity between Escherichia coli in aerosol and Escherichia coli in feces,nasal-mucus and diseased lung tissue samples of pigs were 98.0%—99.2%,94.9%—96.3% and 98.3%—99.1%,respectively.It showed that the Escherichia coli from feces was the main source of Escherichia coli in the aerosol in weaning pig house,Escherichia coli was found in piglet tissues,feces and environment,and the higher homology was possessed between different sources of Escherichia coli.
引文
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[15] 庞广辉,齐莉莉,姜军坡,等.Z-27菌剂对猪舍有害气体排放及育肥猪生长性能的影响[J].河南农业科学,2018,47(9):144-149.
[16] 冯世文,李军,潘艳,等.广西猪源致病性大肠杆菌对抗菌药物的耐药性变化分析[J].南方农业学报,2017,48(2):350-356.
[17] 吴忆春.羊源O78型大肠杆菌的分离及16S rDNA序列分析[J].中国畜牧兽医,2014,41(5):240-243.
[18] BOSSHARD P P,ABELS S,ZBINDEN R,et al.Ribosomal DNA sequencing for identification of aerobic gram-positive rods in the clinical laboratory (an 18-month evaluation)[J].J Clin Microbiol,2003,41(9):4134-4140.
[19] 方光远,茅慧华,蒋加进,等.不同病例动物源大肠杆菌的分离鉴定及16S rDNA同源性分析[J].中国畜牧兽医,2015,42(12):3126-3132.
[2] CHENG D R,SUN H C,XU J S,et al.PCR detection of virulence factor genesin Escherichia coli isolates from weaned piglets with edema disease and/or diarrhea in China[J].Veterinary Microbiology,2006,115(4):320-328.
[3] HERMANN J R,HOFF S J,MU OZ-ZANZI C,et al.Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols[J].Vet Res,2007,38(1):81-93.
[4] 郑龙龙,朱玲云,陈关雄,等.断奶仔猪腹泻病因分析[J].中国畜牧兽医,2015,42(10):2772-2778.
[5] 邢兰君.断奶仔猪水肿病的诊断与治疗[J].猪业科学,2008(8):32-33.
[6] 赵凤菊,曹东,李井春,等.猪源大肠埃希菌超广谱β-内酰胺酶的检测及其耐药性分析[J].河南农业科学,2017,46(2):111-115.
[7] 陈剑波,韩一超,王树华,等.规模化猪场封闭舍内细菌气溶胶粒度分布研究[J].黑龙江畜牧兽医,2016(9):128-133.
[8] 计徐,郑卫江,邹雪婷,等.断奶前后仔猪和母猪粪便源大肠杆菌耐药性和铜抗性的分析[J].畜牧兽医学报,2015,46(3):482-490.
[9] 唐树霄.江苏部分地区致初生、断奶仔猪腹泻水肿病大肠杆菌的O血清型、毒素型和黏附素型的流行病学调查[D].扬州:扬州大学,2013:41-54.
[10] 薛俊龙,张李俊,王采先,等.山西省鸡致病性大肠杆菌的分离及生物特性分析[J].中国兽医杂志,2008,44(1):45-46.
[11] 韩梅英,刘国华,张映西.鸡舍空气中大肠杆菌测定和分离菌的致病力观察[J].中国兽医杂志,2002,38(1):18-19.
[12] 马帅,乔军,孟庆玲,等.新疆奶牛子宫内膜炎大肠杆菌进化分群及耐药特性研究[J].河南农业科学,2018,47(7):116-123.
[13] 王志宇,王国艳,薛俊龙,等.山西省鸡源性大肠杆菌多重耐药性分析[J].山西农业科学,2018,46(2):272-275.
[14] 徐引弟,张青娴,王治方,等.河南省猪肠外致病性大肠杆菌毒力因子的检测及分布[J].山西农业科学,2018,46(11):1921-1925,1948.
[15] 庞广辉,齐莉莉,姜军坡,等.Z-27菌剂对猪舍有害气体排放及育肥猪生长性能的影响[J].河南农业科学,2018,47(9):144-149.
[16] 冯世文,李军,潘艳,等.广西猪源致病性大肠杆菌对抗菌药物的耐药性变化分析[J].南方农业学报,2017,48(2):350-356.
[17] 吴忆春.羊源O78型大肠杆菌的分离及16S rDNA序列分析[J].中国畜牧兽医,2014,41(5):240-243.
[18] BOSSHARD P P,ABELS S,ZBINDEN R,et al.Ribosomal DNA sequencing for identification of aerobic gram-positive rods in the clinical laboratory (an 18-month evaluation)[J].J Clin Microbiol,2003,41(9):4134-4140.
[19] 方光远,茅慧华,蒋加进,等.不同病例动物源大肠杆菌的分离鉴定及16S rDNA同源性分析[J].中国畜牧兽医,2015,42(12):3126-3132.
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