Mai Farghaly, Michael F Hynes, Mohammad Nazari, Sylvia Lee Checkley, Karen Liljebjelke
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Conjugation assays examined transfer frequency of <i>ICEHs02</i> to <i>H. somni</i> and <i>Pasteurella multocida</i> strains. Polymerase chain reaction assays confirmed the presence of a circular intermediate, ICE-associated core genes, and cargo genes in recipient strains. Susceptibility testing examined <i>ICEHs02</i>-associated resistance phenotypes in recipient strains. Tetracycline and ciprofloxacin induction significantly increased the transfer rates of <i>ICEHs02</i> in vitro. The copy numbers of the circular intermediate of <i>ICEHs02</i> per chromosome exhibited significant increases of ∼37-fold after tetracycline exposure and ∼4-fold after ciprofloxacin treatment. 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引用次数: 0
摘要
整合和共轭元件(ICEs)是一种可自我转移的可移动遗传元件,在细菌间通过水平基因转移传播抗菌素耐药性方面起着重要作用。最近在somni Histophilus临床分离株(ICEHs02)中发现的ICE长度为72 914个碱基对,含有7个预测的抗菌素耐药基因,这些基因对四环素(tetret -tet(H))、氟苯尼醇(floR)、磺胺(Sul2)、氨基糖苷(APH(3″)-Ib、APH(6)-Id、APH(3′)-Ia)和铜(mco)具有耐药性。本研究调查了ICEHs02的宿主范围,评估了抗菌应激源对迁移频率的影响,并考察了ICEHs02获取对宿主的影响。偶联试验检测了ICEHs02向索姆尼嗜血杆菌和多杀性巴氏杆菌菌株转移的频率。聚合酶链反应实验证实在受体菌株中存在环状中间产物、ice相关核心基因和货物基因。药敏试验检测受体菌株中与icehs02相关的耐药表型。四环素和环丙沙星诱导显著提高ICEHs02的体外转移率。在四环素和环丙沙星处理后,每条染色体上ICEHs02环状中间体的拷贝数分别显著增加了~ 37倍和~ 4倍。ICEHs02基因的获得使嗜毒黑蝇转共轭子(TG)的相对适合度降低了28% (w = 0.72±0.04),使嗜毒黑蝇转共轭子(TG)的相对适合度降低了15% (w = 0.85±0.01)。
Examination of the horizontal gene transfer dynamics of an integrative and conjugative element encoding multidrug resistance in Histophilus somni.
Integrative and conjugative elements (ICEs) are self-transferable mobile genetic elements that play a significant role in disseminating antimicrobial resistance between bacteria via horizontal gene transfer. A recently identified ICE in a clinical isolate of Histophilus somni (ICEHs02) is 72 914 base pairs in length and harbours seven predicted antimicrobial resistance genes conferring resistance to tetracycline (tetR-tet(H)), florfenicol (floR), sulfonamide (Sul2), aminoglycosides (APH(3″)-Ib, APH(6)-Id, APH(3')-Ia), and copper (mco). This study investigated ICEHs02 host range, assessed effects of antimicrobial stressors on transfer frequency, and examined effects of ICEHs02 acquisition on hosts. Conjugation assays examined transfer frequency of ICEHs02 to H. somni and Pasteurella multocida strains. Polymerase chain reaction assays confirmed the presence of a circular intermediate, ICE-associated core genes, and cargo genes in recipient strains. Susceptibility testing examined ICEHs02-associated resistance phenotypes in recipient strains. Tetracycline and ciprofloxacin induction significantly increased the transfer rates of ICEHs02 in vitro. The copy numbers of the circular intermediate of ICEHs02 per chromosome exhibited significant increases of ∼37-fold after tetracycline exposure and ∼4-fold after ciprofloxacin treatment. The acquisition of ICEHs02 reduced the relative fitness of H. somni transconjugants (TG) by 28% (w = 0.72 ± 0.04) and the relative fitness of P. multocida TG was decreased by 15% (w = 0.85 ± 0.01).
期刊介绍:
Published since 1954, the Canadian Journal of Microbiology is a monthly journal that contains new research in the field of microbiology, including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.