【摘要】：為了解豬源腸球菌分離株的耐藥特點(diǎn)及其毒力基因分布特征,為腸球菌感染疾病的治療防控提供依據(jù),此次研究從湖南省部分豬場(chǎng)的健康豬腸道內(nèi)及患有流行性腹瀉的病豬腸道內(nèi)分離腸球菌,并對(duì)分離菌株進(jìn)行耐藥性及毒力基因檢測(cè)。1.本實(shí)驗(yàn)一共分離腸球菌115株,其中糞腸球菌11株,屎腸球菌59株,其他腸球菌45株。有72株腸球菌(包含30株屎腸球菌,2株糞腸球菌,40株其他腸球菌)分離自四個(gè)豬場(chǎng)的健康豬群腸道內(nèi)；另外43株腸球菌分離自流行性腹瀉病豬腸道內(nèi),包括29株屎腸球菌,9株糞腸球菌,5株其他腸球菌。2.使用K-B法檢測(cè)1 15株腸球菌的耐藥表型并通過PCR方法檢測(cè)其耐藥基因。結(jié)果顯示：健康豬源腸球菌對(duì)高濃度慶大霉素最敏感為93.06%,其次為替考拉寧和氯霉素,敏感率分別為81.94%和65.28%。流行性腹瀉病豬中分離腸球菌對(duì)四環(huán)素、紅霉素、頭孢噻吩這三種抗生素完全不敏感,除糖肽類抗生素和氨芐西林外,對(duì)其它幾種藥物的敏感率均低于健康豬源腸球菌的敏感率。分離的1 1 5株腸球菌多重耐藥現(xiàn)象嚴(yán)重,有98.3%的腸球菌對(duì)14種抗生素呈三重或三重以上的耐藥,比較分析湖南健康豬源與PEDV豬源腸球菌的多重耐藥情況,發(fā)現(xiàn)PEDV豬源腸球菌比健康豬源的腸球菌呈更多重耐藥的菌株數(shù)比健康豬源的腸球菌多。所有的糞腸球菌為六重及六重以上的耐藥,多重耐藥最為嚴(yán)重。本次對(duì)腸球菌的耐藥性進(jìn)行檢測(cè)發(fā)現(xiàn),豬源腸球菌已對(duì)多種抗生素耐藥,尤其是對(duì)四環(huán)素和紅霉素,幾乎是全部耐藥。各耐藥基因的檢出率分別為tetM (69.6%)、 Aac(6)'/aph(2") (62.6%)、aph(3)'-Ⅲ (70.4%)、ant(6)'-Ⅰ (60%)、ermB (58.2%)、 mefA (0.9%)、 VanB (0.9%), VanA和VanC基因未檢測(cè)到。通過耐藥基因型與耐藥表型比較分析,發(fā)現(xiàn)腸球菌耐藥表型與耐藥基因并不完全一致。3.采用PCR方法對(duì)分離的115株腸球菌的毒力基因進(jìn)行檢測(cè),總共檢出9種毒力基因,分別是cylA、ace、esp、hy1、asal、efaA、gelE、EF0591、EF3314。研究結(jié)果表明健康豬腸道內(nèi)分離的72株腸球菌也含有毒力因子,但其存在量較低,各基因的檢出率低于16%。四個(gè)豬場(chǎng)毒力基因的檢出率有較大差異,豬場(chǎng)C的毒力基因檢測(cè)率最高,豬場(chǎng)B未檢測(cè)到帶有毒力基因的菌株,這可能與養(yǎng)殖場(chǎng)的飼養(yǎng)管理相關(guān)或腸球菌的毒力基因在地域與地域之間存在差異。通過健康豬源腸球菌與流行性腹瀉病豬分離腸球菌之間毒力基因檢測(cè)結(jié)果的比較分析,發(fā)現(xiàn)從流行性腹瀉病豬分離的腸球菌檢測(cè)到的毒力基因菌株數(shù)要多。不同腸球菌攜帶耐藥基因的比較中,糞腸球菌毒力基因的攜帶量要多于其他腸球菌。
[Abstract]:In order to understand the characteristics of drug resistance and the distribution of virulence genes of Enterococcus enterococci isolated strains from pigs, and to provide the basis for the treatment and control of enterococcal infection, In this study, Enterococcus was isolated from the intestinal tract of healthy pigs in some pig farms in Hunan Province and from the intestinal tract of diseased pigs suffering from epidemic diarrhea, and the drug resistance and virulence genes of the isolated strains were detected. 1. A total of 115 strains of Enterococcus faecalis, 59 strains of Enterococcus faecium and 45 strains of other Enterococcus were isolated. 72 strains of Enterococcus (including 30 Enterococcus faecium, 2 Enterococcus faecalis and 40 other Enterococcus faecalis) were isolated from the intestinal tract of healthy pigs in four pig farms. Another 43 strains of Enterococcus were isolated from the intestinal tract of epidemic diarrhea pigs, including 29 strains of Enterococcus faecium, 9 strains of Enterococcus faecalis and 5 strains of other enterococci. The resistance phenotypes of 115 strains of Enterococcus were detected by K-B method and their resistant genes were detected by PCR method. The results showed that the highest sensitivity of Enterococcus from healthy pigs to high concentration of gentamicin was 93.06, followed by teicoplanin and chloramphenicol, and the sensitivity rates were 81.94% and 65.2828%, respectively. Enterococcus isolated from epidemic diarrhea pigs was completely insensitive to tetracycline, erythromycin and cefthiophene, except for glycopeptide antibiotics and ampicillin. The sensitivity to other drugs was lower than that of healthy enterococcus. One hundred and fifteen strains of Enterococcus were found to be resistant to three or more antibiotics, 98.3% of which showed triple or more resistance to 14 antibiotics. The multidrug resistance of Hunan healthy swine and PEDV porcine enterococcus was compared and analyzed. It was found that more strains of enterococcus from PEDV were more resistant than those from healthy pigs. All Enterococcus faecalis are six or more resistant, multi-resistant is the most serious. It was found that Enterococcus suis was resistant to many antibiotics, especially tetracycline and erythromycin. The detectable rate of resistant genes was tetM (69.6%), Aac (6'/ aph (2 ") (62.6%), aph (3)'- 鈪，

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