Yingying Cheng1, Yong Chen1, Yang Liu1, Jingjie Song1, Yunbo Chen2, Tongling Shan3, Yonghong Xiao2, Kai Zhou4. 1. Shenzhen Institute of Respiratory Diseases, Second Clinical Medical College (Shenzhen People's Hospital), Jinan University; the First Affiliated Hospital (Shenzhen People's Hospital), Southern University of Science and Technology, Shenzhen, China. 2. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China. 3. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 4. Shenzhen Institute of Respiratory Diseases, Second Clinical Medical College (Shenzhen People's Hospital), Jinan University; the First Affiliated Hospital (Shenzhen People's Hospital), Southern University of Science and Technology, Shenzhen, China. Electronic address: Kai_Zhou@zju.edu.cn.
Abstract
OBJECTIVES: The aim of this study was to characterise a novel tet(X6)-carrying plasmid detected in a livestock-associated Acinetobacter towneri isolate. METHODS: PCR screening was performed to detect tet(X) variants in livestock-associated Acinetobacter spp. isolates. The tet(X6)-positive isolate was analysed by whole-genome sequencing. Functional cloning was performed to detect the activity of Tet(X6). Antibiotic susceptibility was determined by broth dilution and microbiological degradation assays. Site-directed mutagenesis was performed to identify the role of 23-Ala residue of Tet(X6) in tigecycline resistance. RESULTS: The tet(X6) gene was detected on a 159-kb plasmid (pAT205) carried by a tigecycline-susceptible A. towneri isolate recovered from a swine faecal sample. The genetic context of tet(X6) [ΔISVsa3-tet(X6)-abh-guaA-ISVsa3] is highly similar to that of the reported plasmid-borne tet(X) variants, suggesting that it may represent a common structure mediating the dissemination of plasmid-borne tet(X) genes. Additional resistance genes detected on pAT205 were carried by a Tn6205-like region and a disrupted class 2 integron. Gene expression and microbiological degradation assays consistently suggested that the activity of tet(X6) is weaker than that of tet(X3) and tet(X4). The 23-Ala residue of the first FAD-binding site conferred higher activity to Tet(X6) than the Gly reside conserved in the other plasmid-borne tet(X) variants, indicating that the site might be under selection. CONCLUSION: This study alerts to the silent dissemination possibility of tigecycline resistance mediated by a novel plasmid. Continuous monitoring of plasmid-borne tet(X) is imperative for tackling its dissemination.
OBJECTIVES: The aim of this study was to characterise a novel tet(X6)-carrying plasmid detected in a livestock-associated Acinetobacter towneri isolate. METHODS: PCR screening was performed to detect tet(X) variants in livestock-associated Acinetobacter spp. isolates. The tet(X6)-positive isolate was analysed by whole-genome sequencing. Functional cloning was performed to detect the activity of Tet(X6). Antibiotic susceptibility was determined by broth dilution and microbiological degradation assays. Site-directed mutagenesis was performed to identify the role of 23-Ala residue of Tet(X6) in tigecycline resistance. RESULTS: The tet(X6) gene was detected on a 159-kb plasmid (pAT205) carried by a tigecycline-susceptible A. towneri isolate recovered from a swine faecal sample. The genetic context of tet(X6) [ΔISVsa3-tet(X6)-abh-guaA-ISVsa3] is highly similar to that of the reported plasmid-borne tet(X) variants, suggesting that it may represent a common structure mediating the dissemination of plasmid-borne tet(X) genes. Additional resistance genes detected on pAT205 were carried by a Tn6205-like region and a disrupted class 2 integron. Gene expression and microbiological degradation assays consistently suggested that the activity of tet(X6) is weaker than that of tet(X3) and tet(X4). The 23-Ala residue of the first FAD-binding site conferred higher activity to Tet(X6) than the Gly reside conserved in the other plasmid-borne tet(X) variants, indicating that the site might be under selection. CONCLUSION: This study alerts to the silent dissemination possibility of tigecycline resistance mediated by a novel plasmid. Continuous monitoring of plasmid-borne tet(X) is imperative for tackling its dissemination.