The recent emergence of plasmid-mediated tigecycline resistance genes, tet(X3) and tet(X4), in animals and humans in China would pose a foreseeable threat to public health. To illustrate this paradigm shift in tigecycline resistance, here, covering the period 2008-2018, we retrospectively analysed a national strain collection of Escherichia coli (n = 2254), obtained from chickens and pigs, in six representative provinces of China. The gene tet(X4) was identified in five pig isolates collected in 2016 and 2018 from the provinces of Sichuan (3/15, 2018), Henan (1/25, 2018) and Guangdong (1/28, 2016), but not in the isolates prior to 2016. None of the isolates was detected harbouring tet(X3). All tet(X4)-positive E. coli exhibited high levels of tigecycline resistance (MICs, 16-64 mg/L), and two were confirmed as colistin resistant, harbouring chromosome-borne mcr-1 gene. The gene tet(X4) was detected on a plasmid in all five isolates, whereas a co-location of tet(X4) on the chromosome of one isolate was observed. Diverse host strains and novel plasmids related to the tet(X4) gene were observed. Our timely findings of the recent emergence of tet(X4) gene in food animal support the rapid surveillance and eradication of this gene before it is established.
Public health interventions to control the recent emergence of plasmid-mediated tigecycline resistance genes rely on a comprehensive understanding of its epidemiology and distribution over a wide range of geographical scales. Here we analysed an Escherichia coli collection isolated from pigs and chickens in China in 2018, and ascertained that the tet(X4) gene was not present at high prevalence across China, but was highly endemic in northwestern China. Genomic analysis of tet(X4)-positive E. coli demonstrated a recent and regional dissemination of tet(X4) among various clonal backgrounds and plasmids in northwestern China, whereas a parallel epidemic coincided with the independent acquisition of tet(X4) in E. coli from the remaining provinces. The high genetic similarity of tet(X4)-positive E. coli and human commensal E. coli suggests the possibility of its spreading into humans. Our study provides a systematic analysis of the current epidemiology of tet(X4) and identifies priorities for optimising timely intervention strategies.
Brucella melitensis is a facultative intracellular bacterium that replicates within macrophages. The ability of brucellae to survive and multiply in the hostile environment of host macrophages is essential to its virulence. The RNA-binding protein Hfq is a global regulator that is involved in stress resistance and pathogenicity. Here we demonstrate that Hfq is essential for stress adaptation and intracellular survival in B. melitensis. A B. melitensis hfq deletion mutant exhibits reduced survival under environmental stresses and is attenuated in cultured macrophages and mice. Microarray-based transcriptome analyses revealed that 359 genes involved in numerous cellular processes were dysregulated in the hfq mutant. From these same samples the proteins were also prepared for proteomic analysis to directly identify Hfq-regulated proteins. Fifty-five proteins with significantly affected expression were identified in the hfq mutant. Our results demonstrate that Hfq regulates many genes and/or proteins involved in metabolism, virulence, and stress responses, including those potentially involved in the adaptation of Brucella to the oxidative, acid, heat stress, and antibacterial peptides encountered within the host. The dysregulation of such genes and/or proteins could contribute to the attenuated hfq mutant phenotype. These findings highlight the involvement of Hfq as a key regulator of Brucella gene expression and facilitate our understanding of the role of Hfq in environmental stress adaptation and intracellular survival of B. melitensis.
The recently discovered colistin resistance element, mcr-1, adds to the list of antimicrobial resistance genes that rapidly erode the antimicrobial efficacy of not only the commonly used antibiotics but also the last-line agents of carbapenems and colistin. This study investigated the prevalence of the mobile colistin resistance determinant mcr-1 in Salmonella strains recovered from clinical settings in China and the transmission potential of mcr-1-bearing mobile elements harbored by such isolates. The mcr-1 gene was recoverable in 1.4% of clinical isolates tested, with the majority of them belonging to Salmonella enterica serotype Typhimurium. These isolates exhibited diverse pulsed-field gel electrophoresis (PFGE) profiles and high resistance to antibiotics other than colistin and particularly to cephalosporins. Plasmid analysis showed that mcr-1 was carried on a variety of plasmids with sizes ranging from ϳ30 to ϳ250 kb, among which there were conjugative plasmids of ϳ30 kb, ϳ60 kb, and ϳ250 kb and nonconjugative plasmids of ϳ140 kb, ϳ180 kb, and ϳ240 kb. Sequencing of representative mcr-1-carrying plasmids revealed that all conjugative plasmids belonged to the IncX4, IncI2, and IncHI2 types and were highly similar to the corresponding types of plasmids reported previously. Nonconjugative plasmids all belonged to the IncHI2 type, and the nontransferability of these plasmids was attributed to the loss of a region carrying partial or complete tra genes. Our data revealed that, similar to the situation in Escherichia coli, mcr-1 transmission in Salmonella was accelerated by various plasmids, suggesting that transmission of mcr-1-carrying plasmids between different species of Enterobacteriaceae may be a common event.
The objectives of this study were to investigate the prevalence and fluoroquinolone resistant
Salmonella
isolated from an integrated broiler chicken supply chain and their molecular characterization. In total, 73
Salmonella
isolates were recovered from a broiler chicken supply chain in Shanghai.
Salmonella
isolates were tested for susceptibility to 11 antimicrobial agents using the broth dilution method and were characterized using pulsed-field gel electrophoresis (PFGE). Then, the
Salmonella
isolates were examined for mutations in quinolone resistance-determining region (QRDR) of
gyrA
,
gyrB
,
parC
, and
parE
, and were screened for plasmid-mediated quinolone resistance (PMQR) genes. Lastly, we sequenced the plasmids carrying
qnrS1
in six
Salmonella
isolates from three sources (two isolated per source). Among 73
Salmonella
isolates, 45 isolates were identified as
S
. Indiana, 24 were
S
. Schwarzengrund, 2 were
S
. Enteritidis, and 2 were
S
. Stanleyville. In addition, high rates of resistance were detected for nalidixic acid (41.1%) and ciprofloxacin (37.0%), while resistance to other test agents was diverse (2.0–100%).
S
. Indiana and
S
. Schwarzengrund isolates from different sources exhibited the same PFGE pattern, suggesting that the
Salmonella
isolates possessed high potential to spread along the broiler chicken supply chain.
gyrA
and
parC
exhibited frequent missense mutations. Moreover,
qnrS1
was the most prevalent PMQR gene in the 73
Salmonella
isolates, and it was found about a new hybrid plasmid. This study concludes a high prevalence of fluoroquinolone resistant
Salmonella
in chicken supply chain, threatening the treatment of
Salmonella
foodborne diseases. In particular, the emergence of a new hybrid plasmid carrying
qnrS1
indicates that the recombination of plasmid carrying resistance gene might be a potential risk factor for the prevention and control strategies of drug resistance.
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