去甲腎上腺素對大腸桿菌耐藥質粒接合轉移的影響
本文選題:去甲腎上腺素 + 大腸桿菌 ; 參考:《南京農業(yè)大學學報》2017年05期
【摘要】:[目的]本研究旨在探討生產中普遍存在的急性應激因素對細菌耐藥基因水平轉移的可能影響。[方法]選取分離自斷奶香豬糞便的2株喹諾酮類敏感型大腸桿菌作為試驗菌株,通過測定D600值比較菌株在添加不同濃度去甲腎上腺素培養(yǎng)液中的生長曲線,確定去甲腎上腺素對于大腸桿菌的最佳促生長濃度;分離自斷奶香豬糞便的7株攜帶質粒喹諾酮類耐藥基因(aac(6')-lb-cr、oqx AB、qnr S)的大腸桿菌,采用濾膜接合法,計算去甲腎上腺素誘導和未誘導的大腸桿菌分離株(供體菌)與大腸桿菌J53(受體菌)的接合轉移率;PCR檢測接合子及供、受體菌質粒介導抗性基因的攜帶情況;采用K-B紙片擴散法和瓊脂稀釋法測定接合子及供、受體菌的多種抗生素最小抑菌濃度(MIC)。[結果]去甲腎上腺素對于大腸桿菌的促生長效果具有濃度依賴性,最佳促生長濃度為100μmol·L-1。7株攜帶質粒介導的喹諾酮類耐藥基因的大腸桿菌中有4株(A、B、C、D)成功獲得接合子,經去甲腎上腺素(100μmol·L-1)誘導后與受體菌的接合轉移率與去甲腎上腺素未誘導組相比均升高,其中A、B、C 3株供體菌極顯著升高(P0.01)。所有接合子均擴增到恩諾沙星3種耐藥基因:aac(6')-lb-cr、oqx AB、qnr S。接合子菌株對6種抗菌藥物的MIC值與大部分供體菌株相同。[結論]去甲腎上腺素對于大腸桿菌的促生長效果具有濃度依賴性,且提高了大腸桿菌分離株耐藥質粒體外接合轉移率。提示:動物養(yǎng)殖生產中應加強應激管理,以減少細菌耐藥基因水平傳播的發(fā)生。
[Abstract]:[objective] to investigate the possible effects of acute stress factors in production on the horizontal transfer of bacterial drug resistance genes. [methods] two strains of Quinolone-sensitive Escherichia coli isolated from weaning pig feces were selected as experimental strains. The growth curves of the strains were compared by measuring the D600 values in the medium supplemented with different concentrations of norepinephrine. To determine the optimal concentration of norepinephrine for the growth of Escherichia coli, and to isolate 7 strains of E. The conjugation transfer rates of norepinephrine induced and uninduced Escherichia coli isolates (donors) and Escherichia coli J53 (receptor bacteria) were calculated. The conjugates and donors were detected by PCR. K-B disk diffusion method and Agar dilution method were used to determine the minimal inhibitory concentration (MIC) of conjugate and donor antibiotics. [results] the effect of norepinephrine on the growth of Escherichia coli was concentration-dependent. The best concentration was 100 渭 mol L-1.7 strain of Escherichia coli carrying plasmid mediated quinolone resistance gene. After induction with norepinephrine (100 渭 mol L ~ (-1), the rate of conjugated transfer with the receptor bacteria was significantly higher than that of the non-norepinephrine induced group, among which the Agnerinic C _ (3) donor strain was significantly higher than that of the norepinephrine group (P _ (0.01). All the conjugates amplified Enrofloxacin 3 kinds of resistance gene: aacanine 6Con -lb-croqx ABN Qnr S. The MIC values of conjugate strains to 6 antimicrobial agents were the same as those of most donor strains. [conclusion] norepinephrine has a concentration-dependent effect on the growth of Escherichia coli, and increases the in vitro conjugation transfer rate of Escherichia coli resistant plasmids. It is suggested that stress management should be strengthened in animal breeding in order to reduce the horizontal transmission of bacterial drug resistance genes.
【作者單位】: 南京農業(yè)大學動物科技學院;
【基金】:國家重點研發(fā)計劃項目(2016YFD0500500) 國家自然科學基金項目(31372321)
【分類號】:S852.612
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