Table_1_Multidrug Resistant Klebsiella pneumoniae ST101 Clone Survival Chain From Inpatients to Hospital Effluent After Chlorine Treatment.DOCX

In this paper we describe the transmission of a multi-drug resistant Klebsiella pneumoniae ST101 clone from hospital to wastewater and its persistence after chlorine treatment. Water samples from influents and effluents of the sewage tank of an infectious diseases hospital and clinical strains collected from the intra-hospital infections, during a period of 10 days prior to wastewater sampling were analyzed. Antibiotic resistant K. pneumoniae strains from wastewaters were recovered on selective media. Based on antibiotic susceptibility profiles and PCR analyses of antibiotic resistance (AR) genetic background, as well as whole-genome sequencing (Illumina MiSeq) and subsequent bioinformatic analyses, 11 ST101 K. pneumoniae strains isolated from hospital wastewater influent, wastewater effluent and clinical sector were identified as clonally related. The SNP and core genome analyses pointed out that five strains were found to be closely related (with ≤18 SNPs and identical cgMLST profile). The strains belonging to this clone harbored multiple acquired AR genes [blaCTX–M–15, blaOXA–48, blaOXA–1, blaSHV–106, blaTEM–150, aac(3)-IIa, aac(6′)-Ib-cr, oqxA10, oqxB17, fosA, catB3, dfrA14, tet(D)] and chromosomal mutations involved in AR (ΔmgrB, ΔompK35, amino acid substitutions in GyrA Ser83Tyr, Asp87Asn, ParC Ser80Tyr). Twenty-nine virulence genes involved in iron acquisition, biofilm and pili formation, adherence, and the type six secretion system – T6SS-III were identified. Our study proves the transmission of MDR K. pneumoniae from hospital to the hospital effluent and its persistence after the chlorine treatment, raising the risk of surface water contamination and further dissemination to different components of the trophic chain, including humans.

本研究阐述了耐多药肺炎克雷伯菌（Klebsiella pneumoniae）ST101克隆株从医院传播至污水处理系统，以及其经氯消毒处理后仍可存活的现象。本研究对某传染病医院污水处理池的进水与出水水样，以及污水处理采样前10天内从医院内感染病例中分离得到的临床菌株进行了分析。采用选择性培养基从污水样本中分离得到耐抗生素肺炎克雷伯菌菌株。基于抗生素敏感性谱、抗生素耐药性（Antibiotic Resistance, AR）遗传背景的PCR分析，以及全基因组测序（Illumina MiSeq）与后续生物信息学分析，研究人员将从医院污水处理进水、出水以及临床科室分离得到的11株ST101肺炎克雷伯菌鉴定为克隆相关株。单核苷酸多态性（Single Nucleotide Polymorphism, SNP）与核心基因组分析结果显示，其中5株菌株亲缘关系极近（单核苷酸差异数≤18，且核心基因组多位点序列分型（core genome Multilocus Sequence Typing, cgMLST）谱完全一致）。该克隆株携带多种获得性抗生素耐药性基因[blaCTX–M–15、blaOXA–48、blaOXA–1、blaSHV–106、blaTEM–150、aac(3)-IIa、aac(6′)-Ib-cr、oqxA10、oqxB17、fosA、catB3、dfrA14、tet(D)]，以及与抗生素耐药性相关的染色体突变（ΔmgrB、ΔompK35、GyrA Ser83Tyr、Asp87Asn、ParC Ser80Tyr氨基酸替换）。本研究还鉴定出29个与铁摄取、生物膜与菌毛形成、黏附相关的毒力基因，以及六型分泌系统III型（Type Six Secretion System III, T6SS-III）。本研究证实了耐多药（Multi-Drug Resistant, MDR）肺炎克雷伯菌从医院传播至医院污水处理出水系统，且经氯消毒处理后仍可存活，这会增加地表水被污染的风险，并可能进一步向包括人类在内的营养链各环节传播扩散。