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Ceftazidime-avibactam (CZA), a novel β-lactam/β-lactamase inhibitor combination, has good antibacterial activity against carbapenem-resistant Enterobacterales (CRE) producing class A and C and some class D carbapenemases, but in recent years, the emergence of CZA-resistant Enterobacterales bacteria is growing. Therefore, rapid, accurate, and timely detection of CZA is necessary for clinical anti-infection treatment. In this study, the rapid ResaCeftazidime-avibactam Enterobacterales NP test was developed; its principle is that metabolically active bacteria (CZA-resistant strains) can change resazurin-PrestoBlue, a viability colorant, from blue to purple or pink in the presence of CZA, whereas CZA-susceptible strains cannot. We used 178 Enterobacterales isolates to evaluate the performance of this test. This test allowed the susceptibility of Enterobacterales to CZA to be detected within 4.5 h with an overall performance of 96% category agreement (CA), 7% major errors (MEs), and 0% very major errors (VMEs). Performance for Escherichia coli included 100% CA and 0% MEs and VMEs. Performance for Klebsiella pneumoniae included 99% CA and 2% MEs and 0% VMEs. Performance for Enterobacter cloacae included 87% CA, 25% MEs, and 0% VMEs. Moreover, this test is both economical ($1.0106 per isolate) and convenient, as it only requires basic laboratory equipment. In a word, the rapid ResaCeftazidime-avibactam Enterobacterales NP test is rapid and feasible, which may provide certain backing for the rapid screening and timely treatment of CZA-resistant strains in the clinic.

Colistin-resistant (Col-R) bacteria are increasing sharply, which poses a serious threat to public health. Thymol is a phenolic compound used for its wide-spectrum antimicrobial activity, while the combination of nontraditional drugs to restore colistin activity is an attractive strategy to treat infections caused by these pathogens. This study showed that thymol could play a synergistic role with colistin against Gram-negative bacteria (GNB), including nonfermenting bacteria and Enterobacteriaceae. According to antimicrobial resistance profiles, most of the colistin-resistant strains we collected showed multidrug-resistant (MDR) phenotypes. The checkerboard method and time-kill curve confirmed the synergistic effect of thymol combined with colistin against Col-R GNB. The synergistic antibiofilm activity of thymol combined with colistin was assessed via crystal violet staining and scanning electron microscopy (SEM) assays. Results showed that compared with a single drug, the combination partially destroyed bacterial cells and inhibit the formation of bacterial biofilms. Mechanismly, the thymol/colistin combination synergistically potentiated the antibacterial activity by accelerating the damage and permeability of the bacterial outer membrane. Preliminary data indicated that the thymol/colistin combination could decrease the number of bacteria ≥2 log 10 CFU/mL after 24 h of therapy in a mouse thigh infection model. Our results fully prove that thymol and colistin combination possesses a promising treatment option against colistin-resistant GNB infections. IMPORTANCE Colistin is being considered "the last ditch" treatment in many infections caused by multidrug-resistant GNB clinical isolates, but colistin-resistant (Col-R) strains with different drug resistance mechanisms have appeared worldwide. Hence, it is of great significance to rejuvenate sensitization of clinical Col-R Gram-negative bacteria to colistin. In this study, the thymol/colistin combination showed notable antibacterial activity in vitro and in vivo . These findings suggest that the thymol/colistin combination may have promise as a treatment approach for treating the infections caused by Col-R pathogens.

Introduction: Globally, Pseudomonas aeruginosa (PA) is emerging as a predominant nosocomial pathogen that often induces aggressive and even deadly infections. Pseudomonas type III repressor A (PtrA) can be activated specifically by copper ions and interacts with type-III transcriptional activator ExsA. This study aims to provide insight into the PtrA-mediated regulation of the pathogenicity and antibiotics resistance of PA., Methods and Results: The results of transcriptome sequencing analyses and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) showed that PtrA plays a dual regulatory role in the virulence systems of PA: negatively regulates the type-III secretion system (T3SS) and positively regulates the quorum-sensing system (QS). The ptrA mutant attenuated extracellular virulence related to QS like pyocyanin, elastase, rhamnolipids, proteolytic activity, and biofilm production. According to adhesion and invasion experiments, PtrA can not only contribute to the adhesiveness but also the invasive of PA. Moreover, the PtrA-mediated regulation of PA pathogenicity was determined both in vivo and in vitro through cytotoxicity and Galleria mellonella survival experiments. In addition, apart from virulence, PtrA was found to influence the carbapenems resistance of PA. After deleting ptrA , the minimum inhibitory concentration (MIC) of carbapenems antibiotics was decreased by 2-fold, while a 2-8 fold increase was noted for the complemented strain., Conclusion: Our findings establish that PtrA exerts a regulatory role in both pathogenicity and carbapenems resistance of PA. This work may shed light on a novel target for the clinical treatment of PA., Competing Interests: The authors report no conflicts of interest in this work., (© 2022 Zhang et al.)

Colistin is used as the "last line of defense" against multidrug-resistant (MDR) Gram-negative bacteria (GNB). However, improper use of colistin may further lead to an increasing number of colistin-resistant (Col-R) strains worldwide, which greatly limits antibiotic treatment options. In this study, we investigated the antibacterial and antibiofilm activities of naringenin (NG) combined with colistin against Col-R GNB in vitro and in vivo . The checkerboard method and time-kill test showed that NG combined with colistin has better antibacterial activity (FICI < 0.5) compared with NG and colistin alone. Biofilm formation inhibition tests demonstrated that combining the two drugs could inhibit biofilm formation; scanning electron microscopy (SEM) confirmed that the combination of the two significantly reduces the number of cells in the biofilm compared with the drug alone. The in vivo experiment showed that the combination of NG and colistin can improve the survival rate of the Galleria mellonella ( G. mellonella ) and reduce the microbial load in the mouse thigh infection model. Mechanistically, the combination of NG and colistin synergistically enhances the antibacterial activity and changes the permeability of the bacterial outer membrane. More importantly, cytotoxicity tests showed no cell cytotoxicity of NG in combination with colistin. In conclusion, our data revealed that NG combined with colistin exhibited good synergistic effects in vivo and in vitro , thus providing a new therapeutic option for clinical Col-R GNB infections., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xu, Yao, Zhao, Shi, Sun, Feng, Zhou, Zhang, Cao and Zhou.)

Ceftazidime/avibactam (CZA) is an alternative antibiotic used for the treatment of infections caused by carbapenem-resistant Enterobacterales (CRE). However, the CZA-resistant CRE strains have been detected worldwide. Therefore, it is critical to screen CZA-resistant CRE strains in colonized patients or a specific population so as to rapidly implement infection control measures to limit their transmission. In this study, we developed a Salmonella - Shigella (SS) CZA-selective medium and assessed its performance to screen for clinical CZA-resistant CRE isolates in both pure-strain specimens and stool samples. A total of 150 non-duplicated isolates, including 75 CZA-susceptible and 75 CZA-resistant CRE pathogens, were tested by using the broth microdilution method and the SS CZA medium, respectively. The bacterial suspensions were serially diluted in the SS CZA medium, which showed excellent screening performance in both pure CZA-resistant CRE strain and the stool samples with the lowest detection limit of 10 1 -10 2 and 10 1 -10 3 CFU/ml, respectively. Notably, none of the susceptible isolates showed growth even at the highest dilution concentration of 10 8 CFU/ml. Most importantly, the SS CZA medium demonstrated excellent performance in screening simulated clinical polymicrobial specimens. Moreover, its screening performance was unaffected by the different resistance determinants for tested isolates. Cumulatively, our data suggest that the SS CZA medium can be used as a promising selective medium to screen CZA-resistant CRE, irrespective of their resistance mechanisms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zeng, Liao, Zhao, Wang, Shu, Jia, Chen, Zhang, Zhou and Wu.)

Daptomycin is a last-line antibiotic used in the treatment of multidrug-resistant Enterococcus faecium infections. Alarmingly, daptomycin-resistant E. faecium isolates have emerged. In this study, we investigated the evolution and mechanisms of daptomycin resistance in clinical E. faecium isolates and the corresponding acquisition of collateral sensitivity (CS) as an evolutionary trade-off. We evolved daptomycin resistance in six daptomycin-susceptible E. faecium isolates to obtain daptomycin-resistant mutants. The six E. faecium strains successfully acquired high-level resistance to daptomycin in vitro , but this led to fitness costs in terms of growth, in vitro competition, and virulence. Mutations in liaFSR , yycFG , and cls ; increased surface positive charge; thicker cell walls; and elevated expression of dltABCD and tagGH were observed in daptomycin-resistant mutants. Surprisingly, we observed the emergence of CS in SC1762 isolates after the induction of daptomycin resistance. Compared with parental strains, the SC1174-D strain (i.e., daptomycin-resistant mutant of SC1174; non-CS) showed significantly upregulated expression of the vanA gene cluster. However, in SC1762-D (i.e., daptomycin-resistant mutant of SC1762), all vanA cluster genes except the vanX gene were obviously downregulated. Further in silico analyses revealed that an IS 1216E- based composite transposon was generated in SC1762-D, and it disrupted the vanH gene, likely affecting the structure and expression of the vanA gene cluster and resulting in resensitization to glycopeptides. Overall, this study reports a novel form of CS between daptomycin and glycopeptides in E. faecium . Further, it provides a valuable foundation for developing effective regimens and sequential combinations of daptomycin and glycopeptides against E. faecium ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zeng, Feng, Qian, Chen, Wang, Zhang, Zheng, Wang, Liu, Zhou and Sun.)

Objectives: This study investigated the underlying mechanism of the evolution of tigecycline resistance during treatment in a patient infected with Klebsiella pneumoniae harbouring bla KPC-2 ., Methods: A total of seven clonal K. pneumoniae strains were continuously isolated from a patient during hospitalisation. Antimicrobial resistance in the strains was determined by antimicrobial susceptibility testing. Multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were performed to explore the homology of the isolates. Whole-genome shotgun (WGS) analysis and cloning experiments were used to investigate the underlying mechanism of the evolution of tigecycline resistance., Results: All of the isolates had a minimum inhibitory concentration (MIC) for tigecycline of 4 µg/mL, except strain FK6768 that had a MIC of 32 µg/mL. Carbapenem-resistant K. pneumoniae strains (FK6614, FK6768 and FK6809) were consecutively isolated from faeces at different times. Antimicrobial susceptibility testing indicated that tigecycline resistance increased in FK6768 and subsequently decreased in FK6809, which attracted our attention. WGS and further bioinformatics analysis showed a homology for the three faecal isolates of >99%. The bla KPC-2 carbapenemase gene and a tet(A) mutation were found in tigecycline-resistant isolate FK6768. Subsequent cloning experiments confirmed the contribution of a tet(A) variant to reduced tigecycline susceptibility., Conclusion: Here we report a K. pneumoniae isolate carrying both tet(A) mutation and the bla KPC-2 gene, which led to increased tigecycline resistance during tigecycline treatment. This is the first report describing tigecycline resistance of K. pneumoniae first increasing and subsequently decreasing in vivo., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The development of biocompatible materials with desired functions is essential for tissue engineering and biomedical applications. Hydrogels prepared from these materials represent an important class of soft matter for mimicking extracellular environments. In particular, dynamic hydrogels with responsiveness to environments are quite appealing because they can match the dynamics of biological processes. Among the external stimuli that can trigger responsive hydrogels, light is considered as a clean stimulus with high spatiotemporal resolution, complete bioorthogonality, and fine tunability regarding its wavelength and intensity. Therefore, photoresponsiveness has been broadly encoded in hydrogels for biological applications. Moreover, light can be used to initiate gelation during the fabrication of biocompatible hydrogels. Here, we present a critical review of light manipulation tools for the fabrication of hydrogels and for the regulation of physicochemical properties and functions of photoresponsive hydrogels. The materials, photo-initiated chemical reactions, and new prospects for light-induced gelation are introduced in the former part, while mechanisms to render hydrogels photoresponsive and their biological applications are discussed in the latter part. Subsequently, the challenges and potential research directions in this area are discussed, followed by a brief conclusion. STATEMENT OF SIGNIFICANCE: Hydrogels play a vital role in the field of biomaterials owing to their water retention ability and biocompatibility. However, static hydrogels cannot meet the dynamic requirements of the biomedical field. As a stimulus with high spatiotemporal resolution, light is an ideal tool for both the fabrication and operation of hydrogels. In this review, light-induced hydrogelation and photoresponsive hydrogels are discussed in detail, and new prospects and emerging biological applications are described. To inspire more research studies in this promising area, the challenges and possible solutions are also presented., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

Gram-negative bacteria utilize secretion systems to export substrates into their surrounding environment or directly into neighboring cells. These substrates are proteins that function to promote bacterial survival: by facilitating nutrient collection, disabling competitor species or, for pathogens, to disable host defenses. Following a rapid development of computational techniques, a growing number of substrates have been discovered and subsequently validated by wet lab experiments. To date, several online databases have been developed to catalogue these substrates but they have limited user options for in-depth analysis, and typically focus on a single type of secreted substrate. We therefore developed a universal platform, BastionHub, that incorporates extensive functional modules to facilitate substrate analysis and integrates the five major Gram-negative secreted substrate types (i.e. from types I-IV and VI secretion systems). To our knowledge, BastionHub is not only the most comprehensive online database available, it is also the first to incorporate substrates secreted by type I or type II secretion systems. By providing the most up-to-date details of secreted substrates and state-of-the-art prediction and visualized relationship analysis tools, BastionHub will be an important platform that can assist biologists in uncovering novel substrates and formulating new hypotheses. BastionHub is freely available at http://bastionhub.erc.monash.edu/., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Background/purpose: Trimethoprim-sulfamethoxazole (TMP-SMZ) is broadly administered to treat multiple infections, and the paucity of effective treatment alternatives for infections caused by Klebsiella pneumoniae has led to a renewed interest in TMP-SMZ. The aim of this study is to evaluate the antibacterial efficacy of TMP-SMZ against K. pneumoniae., Methods: The resistance genes of K. pneumoniae clinical isolates were investigated by PCR, followed by conjugation experiments and multilocus sequence typing., Results: The resistance rate of K. pneumoniae to TMP-SMZ decreased over the collection period from 26.7% (88/330) to 16.9% (56/332). The high carrying rates (173/175, 98.9%) of resistance determinants (sul genes or dfr genes) were the main mechanisms of TMP-SMZ resistance isolates, with sul1 (142/175, 81.1%) and dfrA1 (119/175, 68.0%). Only class 1 integron was detected, the prevalence of which in TMP-SMZ resistant K. pneumoniae was 63.4% (111/175)., Conclusion: These results provided insights into the antimicrobial efficacy of TMP-SMZ against K. pneumoniae, also illustrating the wide distribution of SMZ and TMP resistance genes among resistant K. pneumoniae. Simultaneously, the present study highlights the significance of reasonable administration and effective continued monitoring., (Copyright © 2019. Published by Elsevier B.V.)

In this study, we report a clinical isolate of a carbapenem-, colistin-, and fosfomycin-resistant Escherichia coli isolate DC-3737 co-harboring bla NDM-1 , mcr-1, and fosA3 from an inpatient in China. Antimicrobial susceptibility testing, polymerase chain reaction, multi-locus sequence typing, conjugation experiment, and Southern blot hybridization were performed on E. coli DC-3737 isolated from the wound. Plasmid analysis is presented and the locations of bla NDM-1 , mcr-1, fosA3, and other resistance genes were identified as well. E. coli DC-3737 was resistant to ampicillin, ceftriaxone, ceftazidime, ciprofloxacin, levofloxacin, gentamicin, tobramycin, trimethoprim-sulfamethoxazole, imipenem, meropenem, ertapenem, fosfomycin and colistin, and with intermediate susceptibility to amikacin. It was typed as sequence type 27. The isolate possessed bla NDM-1 , mcr-1, fosA3, bla CTX-M-9 , bla TEM-1 , aac (6')-Ib-cr and sul1 simultaneously. In addition, the mutations in quinolone resistance-determinant regions (QRDRs) such as Ser83Leu and Asp87Asn in gyrA, and Ser80Ile in parC were detected. Conjugation assays revealed that bla NDM-1 , fosA3, sul1, mcr-1, and bla CTX-M-9 genes could successfully transfer their resistance phenotype to E. coli strain J53. Plasmid analysis and Southern hybridization showed that DC-3737 possessed Z-type self-transmissible plasmid bearing bla NDM-1 , fosA3, and sul1. Moreover, mcr-1, bla CTX-M-9 , and bla TEM-1 were located on a ~60-kb IncFIB type self-transmissible plasmid. This is the first report of bla NDM-1 , mcr-1 and fosA3 co-harboring in E. coli in China. Moreover, it is also the first description of the co-harboring of bla NDM and fosA3 in a single Z plasmid in Enterobacteriaceae species. The identification of E. coli DC-3737 co-harboring bla NDM-1 , mcr-1, and fosA3 in this study highlights the need to increase epidemiologic surveillance and the need for new classes of antibiotics to address multidrug-resistant bacteria.

Objectives: Triclosan is usually employed as a disinfectant in a wide range of medical and consumer care products, which may have imposed a selective pressure on bacteria. This study was designed to evaluate the resistance mechanisms of triclosan and molecular epidemiology of triclosan-resistant isolates of Acinetobacter baumannii in Wenzhou, China., Methods: A collection of 626 A. baumannii were isolated from the First Affiliated Hospital of Wenzhou Medical University during 2016-2017 and antimicrobial susceptibility testing of these isolates were performed via agar dilution method. Molecular mechanisms of triclosan resistance, including the existence of mutations in reductase (FabI) were investigated by PCR and sequencing. Furthermore, quantitative RT-PCR was conducted to evaluate the expression levels of the fabI gene and efflux pump genes (adeB, adeG, adeJ, abeM, amvA and abeS) at normal condition and sub-inhibitory concentration of triclosan, and the epidemiological characteristics were analyzed by PFGE and MLST., Results: 2.7% (17/626) of A. baumannii exhibited resistance to triclosan. The FabI mutation Gly95Ser was found in one triclosan resistant strain. The expression of fabI and adeB gene were significant difference between triclosan-resistant and susceptible strains (P < 0.05). The expression of fabI, adeG, adeJ and abeM were increased after triclosan induction. The clones of these resistant isolates were diverse and sporadic., Conclusions: The hyper-expression of fabI was probably the main mechanism of triclosan resistance in this study, and the efflux pump AdeB, AdeG, AdeJ and AbeM might also be related to decreased triclosan susceptibility., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: To characterize the evolutionary pathways of tigecycline (TGC) resistance and alterations in the biological characteristics of hospital-derived Staphylococcus aureus isolates under selective pressure., Methods: Three clinical S. aureus strains and one standard S. aureus strain, ATCC 29213, were used for the in vitro selection of TGC-resistant S. aureus variants with gradient concentrations of TGC. Changes in drug resistance and genetic alterations in resistance-related genes (operon mepRAB and rpsJ) in mutant strains were determined. The efflux inhibitor assay for MepA and the fitness cost, determined by comparing the growth and virulence of parental and mutant strains, were also investigated., Results: Mutants induced in vitro showed a 64- to 128-fold increase in the minimum inhibitory concentration (MIC) of TGC. Substitution mutations were detected in the transcriptional repressor mepR and the efflux pump gene mepA. A K57M amino acid substitution occurred in the ribosomal S10 protein-encoding gene rpsJ. The MICs of TGC in the final mutants were significantly decreased in the presence of efflux pump inhibitors. It was worth noting that growth was unaffected by TGC resistance selection in vitro, with the exception of one strain, and the MICs of other antibiotics and virulence were also unaffected., Conclusions: The evolution of TGC resistance in S. aureus in vitro is associated with a loss-of-function mutation in the efflux pump transcriptional repressor mepR and a missense mutation in the efflux pump-encoding gene mepA. Our work further validated the resistance mechanisms of S. aureus to TGC and reported previously undiscovered mutations., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This study aimed to investigate the characteristics and mechanisms responsible for heteroresistance to colistin in Acinetobactor baumannii clinical isolates from a Chinese teaching hospital. Five hundred and seventy-six nonduplicate A. baumannii clinical isolates isolated from 2014 to 2015 were tested. Colistin heteroresistance was determined using population analysis profiles (PAPs). Susceptibility testing was conducted using the broth microdilution method (BMD). The ability to form biofilm formation was determined using 96-well flat bottom microtiter plates. Time-kill assays were also conducted. PCR and sequencing were used to detect the presence of resistant genes. Expression levels of efflux pump genes were determined by qRT-PCR. LPS analysis was conducted by SDS-PAGE. Lipid A characteristic were determined via MALDI-TOF MS. Nine colistin heteroresistant A. baumannii clinical isolates which were selected by PAPs, exhibited multidrug-resistant phenotypes. The microplate biofilm assay revealed that colistin heteroresistant A. baumannii clinical isolates had weaker biofilm formation capacity than A. baumannii ATCC19606. Colistin-heteroresistant A. baumannii isolates exhibited regrowth after 12 h at 0.5 × MIC, 1 × MIC, and 2 × MIC. The results of PCR and sequencing revealed mutations of lpxACD in some colistin heteroresistant A. baumannii isolates. qRT-PCR also showed that the expressions of efflux pump genes were upregulated in some of the heteroresistant isolates. Our study was the first report of colistin heteroresistant A. baumannii clinical isolates in China. The transition of colistin heteroresistance to resistance should be of concern in future clinical surveillance.

Objective: To evaluate the influence of sub-minimum inhibitory concentrations (MICs) of ciprofloxacin (CIP) on biofilm formation and virulence factors of Escherichia coli clinical isolates., Methods: Sub-MICs of CIP were determined using growth curve experiments. The biofilm-forming capacity of E. coli clinical isolates and E. coli ATCC 25922 treated or untreated with sub-MICs of CIP was assessed using a crystal violet staining assay. The biofilm structure of E. coli isolate was assessed with scanning electron microscopy (SEM). The expression levels of the virulence genes fim, usp, and iron and the biofilm formation genes of the pgaABCD locus were measured using quantification RT-PCR (qRT-PCR) in E. coli isolates and E. coli ATCC 25922., Results: Based on our results, the sub-MICs of CIP were 1/4 MICs. Sub-MICs of CIP significantly inhibited biofilm formation of E. coli clinical isolates and E. coli ATCC 25922 (p<0.01). SEM analyses indicated that the biofilm structure of the E. coli changed significantly after treatment with sub-MICs of CIP. Expression levels of the virulence genes fim, usp, and iron and the biofilm formation genes of the pgaABCD locus were also suppressed., Conclusions: The results revealed that treatment with sub-MICs of CIP for 24h inhibited biofilm formation and reduced the expression of virulence genes and biofilm formation genes in E. coli., (Copyright © 2019 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. All rights reserved.)

Background/purpose: Only limited information is available about the detailed characteristics of qnrD, a plasmid-mediated quinolone resistance (PMQR) gene. This study aimed to understand the distribution of qnrD and the characterization of qnrD-carrying plasmids in Proteeae., Methods: The distribution of qnrD genes was investigated by polymerase chain reaction (PCR) amplification in 203 consecutive nonduplicate clinical isolates of Proteeae collected from inpatients at the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China. The minimum inhibitory concentrations (MICs) of antibiotics were measured by agar dilution method and other PMQR determinants were also determined by PCR. qnrD was positioned via Southern hybridization and the transferability of qnrD-carrying plasmids was achieved by conjugation experiment. The genetic environment of qnrD was investigated by sequencing, and chromosomal polymorphism for qnrD-positive strains was analyzed by pulsed-field gel electrophoresis (PFGE)., Results: Forty strains carried qnrD, showing decreased fluoroquinolone susceptibility or low-level fluoroquinolone resistance. qnrD was encoded on the plasmid of about 2.7 kb or 5.2 kb in length, which cannot be transferred by liquid conjugation or filter mating, but can be successfully transferred by transduction. The transformants showed 62.5-300-fold increases in the MICs of quinolones compared with the recipient. The plasmids carrying qnrD showed a high similarity with that of Providencia spp. and Proteus vulgaris. PFGE analysis demonstrated that these isolates were divergent and not clone related., Conclusion: qnrD could have originated from Proteeae or presented in these bacteria as a reservoir; furthermore, qnrD could be transferred and spread within the same or across different bacterial species if the plasmids acquired mobile elements under antimicrobial selective pressures., (Copyright © 2016. Published by Elsevier B.V.)

Background: Proteeae isolates displaying multidrug-resistance (MDR) are the second most common causes of hospital-associated infections. The aim of this study was to screen class 1-3 integrons and plasmid-mediated quinolone resistance (PMQR) genes in Proteeae isolates from the First Affiliated Hospital of the Wenzhou Medical University., Materials and Methods: 176 Proteeae isolates were collected from clinical specimens of inpatients between January 2011 and December 2013. Susceptibility testing was determined by the agar dilution method. Class 1-3 integrons and PMQR genes were amplified by polymerase chain reaction, and the variable regions of integrons were determined by restriction fragment length polymorphisms., Results: 68.2% Proteeae isolates exhibited MDR phenotypes: 46.6 and 10.8% Proteeae isolates were positive for intI1 and intI2, respectively. The resistance rate of integron-positive isolates to aminoglycosides, fluoroquinolones, and trimethoprim/sulfamethoxazole was significantly higher than integron-negative isolates. Sequence analysis revealed that dfrA1-sat2-aadA1, dfrA1-catB2-sat2-aadA1, and sat2-aadA1 were first detected in Morganella morganii strains isolated from China. PMQR was determined by qnrD in 40 strains (22.7%)., Conclusion: Our results indicate that class 1 and 2 integrons are common among Proteeae isolates. Meanwhile, qnrD are highly prevalent in Proteeae isolated from our hospital., (© 2016 S. Karger AG, Basel.)