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Aim of this study was to investigate the prevalence and genetic environment of the oxazolidinone resistance gene optrA in Streptococcus suis (S. suis) isolates from diseased pigs in China. A total of 178 S. suis isolates were screened for the optrA gene by PCR. The phenotypes and genotypes of optrA -positive isolates were investigated by antimicrobial susceptibility testing, core genome Multilocus Sequence Typing (cgMLST), capsular serotypes determination and whole-genome sequencing (WGS). Fifty-one (28.7%) S. suis isolates were positive for optrA. Phylogenetic analysis indicated that the spread of the optrA among S. suis isolates was primarily due to horizontal transfer. Analysis of S. suis serotypes from diseased pigs revealed substantial diversity. The genetic environment of optrA was complex and diverse and could be divided into 12 different types. Interestingly, we identified a novel integrative and conjugative element ICE Ssu 988S, carrying optrA and erm (T) genes. This is to the best of our knowledge the first report of the optrA and erm (T) co-located on an ICE in S. suis. Our results showed a high prevalence of optrA gene in S. suis isolates in China. Further research is needed to evaluate the importance of ICEs, as they horizontally propagate important clinical resistance genes. [ABSTRACT FROM AUTHOR]

• This study characterized a novel integrative and conjugative element, carrying eight resistance genes and seven IS elements, from a serovar 2 Glaesserella parasuis ST279 isolate. • Tn6743, a novel transposon carrying six resistance genes, was identified. • This study characterized a novel insertion element ISGpa1, which is the first characterized example in G. parasuis. ICE Gpa1804 was identified in the genome of a serovar 2, ST279 isolate EHP1804 carrying eight different resistance genes from 200 Glaesserella parasuis strains isolated from swine with lower respiratory tract infection in seven provinces of China. Susceptibility testing for EHP1804 was determined by broth microdilution, and its genetic profile was determined by whole-genome sequencing. The complete ICE Gpa1804 was analysed by polymerase chain reaction, conjugation assay and bioinformatics tools. The conjugation assay was performed using EHP1804 as the donor and G. parasuis V43 (rifampicin-resistant) as the recipient. ICE Gpa1804 has a size of 71,880 bp and contains 83 genes, including eight resistance genes [ tet (B), bla Rob-1 , aphA1, strA, strB, aac(3)-IId, catA3 and sul2 ]. The conjugation assay showed that ICE Gpa1804 could be transferred to G. parasuis V43 with frequencies of 4.3 × 10−7. To the best of the authors' knowledge, this is the first study to identify a novel integrative and conjugative element (ICE) carrying eight resistance genes and seven insertion sequence (IS) elements from a G. parasuis isolate. Tn 6743 , a novel transposon carrying six resistance genes, was identified. Moreover, IS Gpa1 , a novel IS 256 family insertion element, is the first characterized example of a G. parasuis insertion element. Multiple mobile genetic elements involved in resistance genes were located in chromosomal ICE Gpa1804 , which showed that ICEs may serve as a vital platform for the accumulation of resistance genes. [ABSTRACT FROM AUTHOR]

This study was undertaken to discern the transmission characteristics of mcr-1 and bla CTX-M-type in one multidrug-resistant Escherichia coli LWY24 from chicken in China. The genetic profiles of LWY24 isolate were determined by conjugation, S1-pulsed-field gel electrophoresis, southern blot hybridization, and whole genome sequencing analysis. Meanwhile, co-transfer of plasmids in LWY24 isolate was screened by dual conjugation assays. The LWY24 isolate was identified as ST93, and harbored 3 conjugative plasmids, pLWY24J-3 (bla CTX-M-55 -bearing IncFⅡ), pLWY24J- mcr-1 (mcr-1 -carrying IncI2), and pLWY24J-4 (non-resistance-conferring IncI1), and one nonconjugative plasmid pLWY24 (bla CTX-M-14 -containing IncHI2/IncHI2A). Numerous resistance genes, insertion sequences (especially IS 26), and transposons were found in the 4 plasmids, suggesting that horizontal transmission have occurred by plasmid mating, homologous recombination, and transpositions. Under the selection pressure of cefotaxime and colistin or cefotaxime alone, the mcr-1 -bearing plasmid and the bla CTX-M-55 -harboring plasmid could be co-transferred at a similar frequency, with 8.00 × 10−4 or 9.00 × 10−4 transconjugants per donor cell, respectively. The specific shufflon region in mcr-1 -encoding plasmid could generate up to 6 diverse PilV structures, which may further accelerate the horizontal transfer of plasmid. In conclusion, the transmission characteristics of mcr-1 and bla CTX-M-type in LWY24 isolate could due to clonal spread of ST93, selective pressure of cefotaxime, IS 26 -mediate homologous recombination and transposition, and the specific shufflon region. [ABSTRACT FROM AUTHOR]

Here, the mechanisms of colistin heteroresistance (CHR) were assessed in 12 Escherichia coli isolates from swine in China. CHR was investigated by population analysis profile tests. CHR stability was studied by culturing isolates for five overnight incubation periods in colistin-free medium. Subsequently, the mcr-1 gene and mutations in PmrAB, PhoPQ, and MgrB were screened in parental isolates and resistant subpopulations. Additionally, the expression levels of phoPQ , its target gene pagP , and its negative regulator gene mgrB , as well as pmrAB and its target genes pmrHFIJKLM and pmrC , were determined by real-time relative quantitative PCR. Eleven of the 12 isolates were confirmed to show CHR, with 17 resistant subpopulations. Also, 11 of the 17 subpopulations (64.71%) harbored point mutations in PmrB and/or PhoQ, differing from their parental isolates. However, only one stable resistant subpopulation (EPF42-4) was identified; it harbored an arginine-to-proline substitution at position 93 (R 93 P) within the PmrB HAMP (histidine kinase, adenylyl cyclase, methyl-binding protein, and phosphatase) domain. Compared to the pmrB expression levels in the parental isolate EPF42 and E. coli K-12 MG1655, remarkable pmrB overexpression was observed in EPF42-4, which showed upregulated pmrA , pmrK , and pmrC expression. Structural analysis demonstrated that the R 93 P substitution promotes conformational changes in the HAMP domain, leading to an acceleration in its signal transduction ability and the activation of PmrB expression. In conclusion, point mutations in PmrB and/or PhoQ were primarily associated with CHR. The R 93 P substitution resulted in the establishment of stable resistant subpopulations in E. coli showing CHR., (Copyright © 2020 American Society for Microbiology.)

Background: The plasmid-encoded multidrug efflux pump oqxAB confers bacterial resistance primarily to olaquindox, quinolones, and chloramphenicol. The aims of this study were to investigate the prevalence of oqxAB among Escherichia coli isolates from dogs, cats, and humans in Henan, China and the susceptibilities of E. coli isolates to common antibiotics., Methods: From 2012 to 2014, a total of 600 samples which included 400 rectal samples and 200 clinical human specimens were tested for the presence of E. coli . All isolates were screened for oqxAB genes by PCR and sequencing. The MICs of 11 antimicrobial agents were determined by the broth microdilution method. A total of 30 representative oqxAB -positive isolates were subjected to ERIC-PCR and MLST. Additionally, conjugation experiments and southern hybridizations were performed., Results: Of 270 isolates, 58.5% (62/106) of the isolates from dogs, 56.25% (36/64) of the isolates from cats, and 42.0% (42/100) of the isolates from humans were positive for the oqxAB . Olaquindox resistance was found for 85.7%-100% of oqxAB -positive isolates. Of oqxAB -positive isolates from dogs, cats, and humans, ciprofloxacin resistance was inspected for 85.8%, 59.1%, and 93.8%, respectively. Several oqxAB -positive isolates were demonstrated by ERIC-PCR and MLST, and have high similarity. Phylogenetic analysis showed that oqxAB -positive isolates could be divided into 7 major clusters. OqxAB -positive conjugants were obtained, southern hybridization verified that the oqxAB gene complex was primarily located on plasmids., Conclusion: In conclusion, oqxAB -positive isolates were widespread in animals and humans in Henan, China. Carriage of oqxAB on plasmids of E. coli isolates may facilitate the emergence of multidrug resistant and its transmission via horizontal transfer, and might pose a potential threat to public health., Competing Interests: With regards to our study’s use of animals, this study protocol was reviewed and approved by the Henan Agriculture University animal ethics committee, and the experiment was performed in accordance with the regulations and guidelines established by this committee. For experiments involving human participants, this study was approved by the Chinese Academy of Sciences human ethics committee, and was carried out strictly in accordance with the approved guidelines. Informed consent was obtained from the hospital.Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.