牛源大腸桿菌毒素基因、黏附素基因、O血清型及耐藥性調(diào)查
發(fā)布時間：2018-11-05 16:14
【摘要】：大腸桿菌是大腸桿菌科、埃希氏菌屬的成員,是Escherich在1885年發(fā)現(xiàn)的,一直被認(rèn)為是腸道中的正常菌群,不會致病。到了20世紀(jì)中葉,發(fā)現(xiàn)在動物和人,特別是幼齡動物和嬰兒,大腸桿菌的一些血清型有一定的致病性,往往會引起嚴(yán)重腹瀉和敗血癥,無論在發(fā)達(dá)國家還是在發(fā)展中國家,大腸桿菌導(dǎo)致的腹瀉疾病仍然是一個嚴(yán)重威脅人類健康和導(dǎo)致病畜生長發(fā)育遲緩,生產(chǎn)性能降低,甚至死亡,嚴(yán)重?fù)p害養(yǎng)殖業(yè)的重要因素。本研究針對黑龍江省不同地區(qū)的不同牛場,共采集108頭犢牛腹瀉糞便病料。通過麥康凱培養(yǎng)基與伊紅美藍(lán)培養(yǎng)基的聯(lián)合培養(yǎng),利用16S r RNA通用引物進(jìn)行PCR檢測與序列分析,并結(jié)合傳統(tǒng)大腸桿菌生化試驗鑒定方法,共分離、鑒定出88株大腸桿菌,并對其進(jìn)行了毒素基因(Eae+Stx1+Stx2+bfp A+LT+Sta+Stb+)與黏附素基因(k88+k99+F17+F6+F18+F41+)檢測、O血清型篩選以及耐藥性分析。研究結(jié)果表明:共有38株大腸桿菌檢測出毒素基因,檢出率為43.18%,可分為11種毒素類型,以eae、sta、stb、bfp A4種為主要類型,分別為13株、8株、15株和16株。以基因型為基礎(chǔ),可將這些大腸桿菌分為5種類型:ETEC、非典型EPEC、典型EPEC、ETEC/EPEC以及ETEC/EHEC。其中ETEC共檢出15株,占毒素基因陽性菌株的39.47%。共有60株菌株表達(dá)一種或多種黏附素,檢出率為68.18%。本研究共檢出11種黏附素類型,其中以F18和K88黏附素類型為主,分別占黏附素陽性菌株的53.33%和20%。采用血清凝集實驗進(jìn)行了19種常見致病性O(shè)血清型的檢測,共檢出14種血清型,優(yōu)勢血清型為O2,O125和O128。通過藥敏實驗可知分離菌株呈多重耐藥,對三種或更多抗生素產(chǎn)生耐藥性,其中,對利福平(96.6%)、阿莫西林(97.7%)和氨芐西林(97.7%)表示強耐藥性,對慶大霉素(30.7%)、氧氟沙星(30.7%)、環(huán)丙沙星(33.0%)和萘啶酸(37.5%)較敏感。本研究表明,黑龍江地區(qū)導(dǎo)致犢牛腹瀉的大腸桿菌類型多樣化,其中主要以ETEC,EPEC為主,O血清型分布散在,無集中趨勢,且分離株存在嚴(yán)重的耐藥性。
[Abstract]:Escherichia coli, a member of the family Escherichia coli, was discovered by Escherich in 1885. By the middle of the 20th century, it was found that in animals and humans, especially young animals and infants, some of the serotypes of E. coli were pathogenicity, often leading to severe diarrhoea and septicemia, both in developed and developing countries. The diarrhea disease caused by Escherichia coli is still a serious threat to human health and lead to the growth retardation of diseased animals, production performance decline, even death, serious damage to the aquaculture industry an important factor. In this study, 108 calves were collected from different cattle farms in Heilongjiang province. In this paper, 88 strains of Escherichia coli were isolated by co-culture of wheat Kang Kai medium and Ihloromethylene blue medium, using 16s r RNA universal primer for PCR detection and sequence analysis, and combining with traditional methods of biochemical identification of Escherichia coli. The toxin gene (Eae Stx1 Stx2 bfp A LT Sta Stb) and adhesion gene (k88k99 F 17 F 6 F 18 F 41) were detected, O serotype screening and drug resistance analysis were performed. The results showed that a total of 38 strains of Escherichia coli were detected, and the detection rate was 43.18, which could be divided into 11 types, including 13 strains, 8 strains, 15 strains and 16 strains of eae,sta,stb,bfp A. Based on genotypes, these Escherichia coli can be classified into five types: ETEC, atypical EPEC, typical EPEC,ETEC/EPEC and ETEC/EHEC. Among them, 15 strains were detected by ETEC, accounting for 39.47% of toxin gene positive strains. A total of 60 strains expressed one or more adhesives, and the detection rate was 68. 18%. In this study, 11 types of adhesion were detected, of which F18 and K88 were the main types, accounting for 53.33% and 20% of the positive strains, respectively. Serum agglutination test was used to detect 19 common pathogenic O serotypes. A total of 14 serotypes were detected. The dominant serotypes were O _ 2O _ (125) and O _ (128). The results showed that the isolates were multidrug resistant and resistant to three or more antibiotics, including rifampicin (96.6%), amoxicillin (97.7%) and ampicillin (97.7%). It was sensitive to gentamicin (30.7%), ofloxacin (30.7%), ciprofloxacin (33.0%) and naphthalic acid (37.5%). The results showed that the type of Escherichia coli caused by diarrhea of calves in Heilongjiang province was diversified, in which ETEC,EPEC was the main type, O serotype was scattered, there was no tendency of concentration, and the isolates had serious drug resistance.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.61
本文編號：2312602
[Abstract]:Escherichia coli, a member of the family Escherichia coli, was discovered by Escherich in 1885. By the middle of the 20th century, it was found that in animals and humans, especially young animals and infants, some of the serotypes of E. coli were pathogenicity, often leading to severe diarrhoea and septicemia, both in developed and developing countries. The diarrhea disease caused by Escherichia coli is still a serious threat to human health and lead to the growth retardation of diseased animals, production performance decline, even death, serious damage to the aquaculture industry an important factor. In this study, 108 calves were collected from different cattle farms in Heilongjiang province. In this paper, 88 strains of Escherichia coli were isolated by co-culture of wheat Kang Kai medium and Ihloromethylene blue medium, using 16s r RNA universal primer for PCR detection and sequence analysis, and combining with traditional methods of biochemical identification of Escherichia coli. The toxin gene (Eae Stx1 Stx2 bfp A LT Sta Stb) and adhesion gene (k88k99 F 17 F 6 F 18 F 41) were detected, O serotype screening and drug resistance analysis were performed. The results showed that a total of 38 strains of Escherichia coli were detected, and the detection rate was 43.18, which could be divided into 11 types, including 13 strains, 8 strains, 15 strains and 16 strains of eae,sta,stb,bfp A. Based on genotypes, these Escherichia coli can be classified into five types: ETEC, atypical EPEC, typical EPEC,ETEC/EPEC and ETEC/EHEC. Among them, 15 strains were detected by ETEC, accounting for 39.47% of toxin gene positive strains. A total of 60 strains expressed one or more adhesives, and the detection rate was 68. 18%. In this study, 11 types of adhesion were detected, of which F18 and K88 were the main types, accounting for 53.33% and 20% of the positive strains, respectively. Serum agglutination test was used to detect 19 common pathogenic O serotypes. A total of 14 serotypes were detected. The dominant serotypes were O _ 2O _ (125) and O _ (128). The results showed that the isolates were multidrug resistant and resistant to three or more antibiotics, including rifampicin (96.6%), amoxicillin (97.7%) and ampicillin (97.7%). It was sensitive to gentamicin (30.7%), ofloxacin (30.7%), ciprofloxacin (33.0%) and naphthalic acid (37.5%). The results showed that the type of Escherichia coli caused by diarrhea of calves in Heilongjiang province was diversified, in which ETEC,EPEC was the main type, O serotype was scattered, there was no tendency of concentration, and the isolates had serious drug resistance.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.61
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 劉芳寧,楊增岐,張淑霞,金利鋒,梁元賀;犢牛細(xì)菌性出血性腸炎的病原分離與鑒定[J];動物醫(yī)學(xué)進(jìn)展;2003年02期
2 袁萬哲,何孔旺,陸承平,郝勤宗,裘孝良,左玉柱;產(chǎn)腸毒素性大腸桿菌主要毒力因子的研究進(jìn)展[J];動物醫(yī)學(xué)進(jìn)展;2005年02期
3 王波云,李立安,顧亞夫;細(xì)菌耐藥性研究進(jìn)展與對策[J];解放軍藥學(xué)學(xué)報;2001年05期
4 李俊生,李小麗,周茹;犢牛腹瀉病因探討及治療[J];陜西農(nóng)業(yè)科學(xué);2004年06期
5 范書才,郭景煜;犢牛和羔羊腹瀉大腸桿菌F_(17)抗原菌株的研究[J];中國獸醫(yī)雜志;1988年06期
相關(guān)博士學(xué)位論文 前1條
1 成大榮;斷奶仔豬源大腸桿菌毒力因子的分子流行病學(xué)及F18菌毛部分特性的研究[D];揚州大學(xué);2005年
相關(guān)碩士學(xué)位論文 前1條
1 陳先亮;江蘇某地區(qū)部分牛場產(chǎn)志賀毒素大腸桿菌的分離鑒定及其遺傳相關(guān)性分析[D];揚州大學(xué);2012年
，本文編號：2312602
本文鏈接：http://sikaile.net/yixuelunwen/dongwuyixue/2312602.html
最近更新
教材專著
