Liu Yingying, Qiao Zhu, Ma Yan, Wang Mingcheng, Hu Gongzheng, Li Enzhong
School of Biological and Food Processing Engineering, Huanghuai University, Zhumadian, China.
College of Animal Husbandry and Veterinary Science, Henan Agricultural University, Zhengzhou, China.
Front Vet Sci. 2024 Oct 23;11:1430398. doi: 10.3389/fvets.2024.1430398. eCollection 2024.
Here, we report the genetic features and evolutionary mechanisms of two (M)-bearing plasmids (pTA2 and pTA7) recovered from swine isolates. The genetic profiles of pTA2 and pTA7 and corresponding transconjugants were accessed by S1 nuclease pulsed-field gel electrophoresis and Southern hybridization, followed by whole genome sequencing and bioinformatics analysis. The biological influences of pTA2 and pTA7 were determined by stability and direct competition assays. Both pTA7 and pTA2 had the IncR backbone sequences but differed in the multidrug resistance region (MDR). The MDR of pTA2 consisted of , (M), S1, , , , , , , and (A)- (A) in addition to 22 insertion sequences. Notably, pTA2 carried the novel complex Tn (IS-ctp-lp- (M)-hp-IStnp-IntI4-IS) harboring (M). The fragment carrying (M) (ISlp- (M)-IS---Int), named Tn, and the two resistance modules IS--- and (A)- (A) were located in the MDR of pTA7. Both pTA2 and pTA7 were highly stable in DH5α cells with no fitness cost to the host or disadvantage in growth competition. These results indicate that transposons carrying (M) continuously integrate via mediation with an insertion sequence, which accelerates the transmission of (M) in isolates through integration of other drug-resistant genes, thereby posing a potential serious threat to the efficacy of clinical treatment.
在此,我们报告了从猪分离株中获得的两个携带(M)的质粒(pTA2和pTA7)的遗传特征和进化机制。通过S1核酸酶脉冲场凝胶电泳和Southern杂交,随后进行全基因组测序和生物信息学分析,对pTA2和pTA7以及相应的转接合子的遗传图谱进行了分析。通过稳定性和直接竞争试验确定了pTA2和pTA7的生物学影响。pTA7和pTA2均具有IncR主干序列,但在多药耐药区域(MDR)有所不同。pTA2的MDR除了22个插入序列外,还包括、(M)、S1、、、、、、和(A)-(A)。值得注意的是,pTA2携带了含有(M)的新型复合转座子Tn(IS-ctp-lp-(M)-hp-IStnp-IntI4-IS)。携带(M)的片段(ISlp-(M)-IS---Int),命名为Tn,以及两个抗性模块IS---和(A)-(A)位于pTA7的MDR中。pTA2和pTA7在DH5α细胞中都高度稳定,对宿主没有适应性代价,在生长竞争中也没有劣势。这些结果表明,携带(M)的转座子通过插入序列的介导不断整合,通过整合其他耐药基因加速了(M)在分离株中的传播,从而对临床治疗效果构成潜在的严重威胁。