Characteristics and Epidemiology of Extended-Spectrum beta-Lactamase-Producing Multidrug-Resistant Klebsiella pneumoniae From Red Kangaroo, China
FRONTIERS IN MICROBIOLOGY
Authors: Wang, Xue; Kang, Qian; Zhao, Jianan; Liu, Zhihui; Ji, Fang; Li, Junbao; Yang, Jianchun; Zhang, Chenglin; Jia, Ting; Dong, Guoying; Liu, Shelan; Hu, Guocheng; Qin, Jianhua; Wang, Chengmin
Abstract
Due to its drug resistant nature, beta-lactamase represents a serious challenge for public health. Extended-spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae clones are increasingly reported worldwide. Little is known about the prevalence and biological characteristics of drug-resistant strains in zoos. During routine surveillance at the Zhengzhou Zoo of China, we found Klebsiella pneumoniae isolate in healthy Red Kangaroos (Macropus Rufus) with severe MDR. The Klebsiella pneumoniae were especially resistant to Cefuroxime Sodium (MIC, > 64 mu g/mL), Ceftriaxone (MIC, >8 mu g/mL) and Cefepime (MIC, >64 mu g/mL), and belonged to ST290. Subsequently, whole genome sequencing (WGS) showed that the Chrome Chr-M297-1 harbored bla(DHA-3), bla(SHV-1), bla(CTX-M-14), fosA5, dfrA3, sul3, etc., and pM297-1.1 [222,864 bp, IncFIB(K)], which carried nine antimicrobial genes including bla(CTX-M-14), bla(TEM-191), aph(3 '')-Ib, aph(6)-Id and qnrS1, etc., and pM297-1.2 [225,763 bp, IncFII(K)] carried 22 antimicrobial genes including bla(TEM-1), bla(CTX-M-3), aph(3 ')-Ia, aac(3)-IIa, aac(6 ')-Ib-cr, aadA16, qnrB2, qnrS1, qacE Delta 1, mphA, sul1, and dfrA27, etc. A traceability analysis then revealed that these two plasmids were highly similar to those recovered from human clinical samples in some southern cities in Sichuan Province, China (>99%), suggesting that these plasmids are spreading in China. Furthermore, two plasmids harboring conjugal transfer genes facilitated the transmission of antimicrobial genes by conjugation with E. coli J53. Our research shows that the transmission and adaptation of Klebsiella pneumoniae producing ESBLs is occurring in zoo environments, suggesting that zoos may be becoming important potential reservoirs for clinically important drug-resistant genes. It is therefore necessary to monitor the emergence and spread of drug-resistant gene strains in captive wild animals held in zoo environments.
Integration of the pSLT Plasmid into the Salmonella Chromosome Results in a Temperature-Sensitive Growth Defect Due to Aberrant DNA Replication
JOURNAL OF BACTERIOLOGY
Authors: Wozniak, Christopher E.; Hendriksen, Jordan J.; Olivera, Baldomero M.; Roth, John R.; Hughes, Kelly T.
Abstract
A mutant of Salmonella enterica serovar Typhimurium was isolated that simultaneously affected two metabolic pathways as follows: NAD metabolism and DNA repair. The mutant was isolated as resistant to a nicotinamide analog and as temperature-sensitive for growth on minimal glucose medium. In this mutant, Salmonella's 94-kb virulence plasmid pSLT had recombined into the chromosome upstream of the NAD salvage pathway gene pncA. This insertion blocked most transcription of pncA, which reduced uptake of the nicotinamide analog. The pSLT insertion mutant also exhibited phenotypes associated with induction of the SOS DNA repair system, including an increase in filamentous cells, higher exonuclease III and catalase activities, and derepression of SOS gene expression. Genome sequencing revealed increased read coverage extending out from the site of pSLT insertion. The two pSLT replication origins are likely initiating replication of the chromosome near the normal replication terminus. Too much replication initiation at the wrong site is probably causing the observed growth defects. Accordingly, deletion of both pSLT replication origins restored growth at higher temperatures. IMPORTANCE In studies that insert a second replication origin into the chromosome, both origins are typically active at the same time. In contrast, the integrated pSLT plasmid initiated replication in stationary phase after normal chromosomal replication had finished. The gradient in read coverage extending out from a single site could be a simple but powerful tool for studying replication and detecting chromosomal rearrangements. This technique may be of particular value when a genome has been sequenced for the first time to verify correct assembly.