Plasmid-mediated Quinolone Resistance Determinants Research Articles

Investigation of plasmid-mediated quinolone resistance genes among clinical isolates of Klebsiella pneumoniae in southwest Iran.

BackgroundExtensive and inappropriate use of quinolones has led to growing resistance rates to these broad‐spectrum antibiotics. The present study purposed to investigate the prevalence of plasmid‐mediated quinolone resistance (PMQR) genes in Klebsiella pneumoniae clinical isolates.MethodNinety‐two non‐repetitive K. pneumoniae clinical isolates were confirmed by standard microbiological methods. Antibacterial susceptibility of isolates toward seven agents from the quinolone family was evaluated by the disc diffusion method. Ciprofloxacin minimum inhibitory concentrations (MICs) were determined using the standard agar dilution method. PCR amplification was used to detect the existence of PMQR genes in the studied isolates.ResultsIn the present study, significant quinolones' resistance (40%) was observed in K. pneumoniae isolates, and most of the strains were resistant to nalidixic acid (94.6%) and ofloxacin (45.6%). MIC analysis showed 15 strains were resistant to 6–128 μg/ml of ciprofloxacin, and five were intermediately‐resistant. PMQR genes were detected in 88% of all isolates. Acc(6’)‐Ib‐cr was constituted half of the total PMQR genes detected among ciprofloxacin non‐susceptible isolates. Of 20 ciprofloxacin non‐susceptible isolates, 65% (n = 13) harbored multiple PMQR determinants, and 15 strains were determined as integron carriage.ConclusionThe findings of this study indicated considerable resistance against quinolones, which could be correlated with the extensive and inappropriate use of this class of antibiotics as empirical treatment.

Plasmid-Mediated Fluoroquinolone Resistance in Pseudomonas aeruginosa and Acinetobacter baumannii.

Introduction Pseudomonas aeruginosa and Acinetobacter baumannii are important pathogens in health care–associated infections. Fluoroquinolone resistance has emerged in these pathogens. In this study, we aimed to determine the occurrence of plasmid-mediated quinolone resistance (PMQR) determinants ( qnrA , qnrB , qnrS , aac(6′)-Ib-cr , oqxAB , and qepA ) by polymerase chain reaction (PCR) and the transmissibility of plasmid-borne resistance determinants in clinical isolates of P. aeruginosa and A. baumannii . Materials and Methods The study included P. aeruginosa (85) and A. baumannii (45) which were nonduplicate, clinically significant, and ciprofloxacin resistant. Antibiotic susceptibility testing was done by disk diffusion method for other antimicrobial agents, namely amikacin, ceftazidime, piperacillin/tazobactam, ofloxacin, levofloxacin, and imipenem. Minimum inhibitory concentration of ciprofloxacin was determined. Efflux pump activity was evaluated using carbonyl-cyanide m-chlorophenylhydrazone (CCCP). The presence of PMQR genes was screened by PCR amplification. Transferability of PMQR genes was determined by conjugation experiment, and plasmid-based replicon typing was performed. Results Resistance to other classes of antimicrobial agents was as follows: ceftazidime (86.9%), piperacillin/tazobactam (73.8%), imipenem (69.2%), and amikacin (63.8%). The minimal inhibitory concentration (MIC)50 and MIC90 for ciprofloxacin were 64 and greater than or equal to 256 µg/mL, respectively. There was a reduction in MIC for 37 (28.4%) isolates with CCCP. In P. aeruginosa , 12 (14.1%) isolates harbored qnrB , 12 (14.1%) qnrS , 9 (10.5%) both qnrB and qnrS , 66 (77.6%) aac(6′)-Ib-cr , and 3 (3.5%) oqxAB gene. In A. baumannii , qnrB was detected in 2 (4.4%), 1 (2.2%) harbored both the qnrA and qnrS , 1 isolate harbored qnrB and qnrS , 21 (46.6%) aac(6′)-Ib-cr , and 1 (2.2%) isolate harbored oqxAB gene. Notably, qepA gene was not detected in any of the study isolates. Conjugation experiments revealed that 12 (9.2%) were transferable. Of the transconjugants, seven (58.3%) belonged to IncFII type plasmid replicon, followed by four (33.3%) IncA/C and one (8.3%) IncFIC type. Conclusion The plasmid-mediated resistance aac(6′)-Ib-cr gene is primarily responsible for mediating fluoroquinolone resistance in clinical isolates of P . aeruginosa and A. baumannii . The predominant plasmid type is IncFII.

Carriage of plasmid-mediated qnr determinants and quinolone efflux pump (qepA) by ciprofloxacin-resistant bacteria recovered from Urinary Tract Infection (UTI) samples

BackgroundBacterial resistance to commonly-used antibiotics has been on the increase especially in the clinical settings. This study focused on the detection of plasmid-mediated quinolone resistance (PMQR) determinants in ciprofloxacin-resistant bacteria recovered from Urinary Tract Infection (UTI) samples.ResultsAlready characterized isolates from urine samples of UTI-diagnosed in- and out- patients were obtained from the culture pool of the Department of Medical Microbiology and Parasitology, University College Hospital (UCH), Ibadan, Nigeria. A total of seventy-three ciprofloxacin-resistant isolates were used in this study. Of the 73 UTI isolates, 43 carried at least one of the four PMQR genes targeted and they belonged to eight bacterial genera namely: Escherichia (25), Klebsiella (10), Pseudomonas (2), Proteus (2) and one isolate each belonging to Enterobacter, Acinetobacter, Citrobacter and Salmonella genera. qnrA was detected in 10.9% (8/73) of the isolates while the occurrence of qnrB and qnrS was 32.9% (24/73) and 20.5% (15/73), respectively. The quinolone efflux pump (qepA) was detected in 9/73 (12.3%) of the isolates. Thirty of the isolates carried only one PMQR gene, while thirteen carried two PMQR genes. There was no carriage of more than two PMQR genes in the forty-three isolates from which PMQR genes were detected.ConclusionThis study reports the carriage of PMQR determinants by eight of the nine Gram-negative bacterial genera from urinary sources in patients attending the University College Hospital, Ibadan over the four-month period of study. This is quite worrisome as it suggests a high contribution of UTI cases to the burden of quinolone resistance. There is a need for more studies of this nature in other hospitals in Nigeria, to develop a database on the contribution of UTI cases to quinolone resistance.

Plasmid-mediated quinolone resistance determinants in clinical bacterial pathogens isolated from the Western Region of Ghana: a cross-sectional study.

quinolones are critically important antibiotics that are reserved for treating very severe infections caused by multidrug-resistant bacterial pathogens. However, their indiscriminate uses have resulted in an increased number of resistant strains in many parts of the world including Ghana. We determined the quinolone resistance profile of Gram-negative bacterial pathogens and characterized the underlying molecular determinants of resistance. Gram-negative pathogens obtained from clinical specimens at three hospital laboratories were tested for resistance to quinolones and other commonly used antibiotics. ESBL production among the Enterobacterial isolates was confirmed using the combined disc diffusion method. We then used PCR to determine seven types of plasmid-mediated quinolone resistance genes present in the isolates resistant to nalidixic acid and ciprofloxacin. in this study, 29.5% of the isolates were resistant to ciprofloxacin, with the highest of 50% among E. coli resistance to the other quinolones was levofloxacin (24.4%), norfloxacin (24.9%), and nalidixic acid (38.9%). Significant proportions of the quinolone-resistant isolates were ESBL producers (P-values < 0.001). The aac(6´)-Ib-cr, qnrS, oqxA, and qepA genes were present in 43 (89.6%), 27 (56.3%), 23 (47.9%), and one (2.1%) of the isolates, respectively. None of the isolates tested positive to qnrA, qnrB, and oqxB genes. The presence of the aac(6´)-Ib-cr gene positively correlated with resistance to ceftriaxone, cefotaxime, and gentamicin (P-values < 0.05). high proportions of Gram-negative bacterial isolates were resistant to quinolones and most of these isolates possessed multiple PMQR genes. There is a need to implement measures to limit the spread of these organisms.

The molecular mechanisms of fluoroquinolone resistance found in rectal swab isolates of Enterobacterales from men undergoing a transrectal prostate biopsy: the rationale for targeted prophylaxis

BackgroundTransrectal ultrasound-guided prostate biopsy (TRUS-Bx) is considered an essential urological procedure for the histological diagnosis of prostate cancer. It is, however, considered a “contaminated” procedure which may lead to infectious complications. Recent studies suggest a significant share of fluoroquinolone-resistant rectal flora in post-biopsy infections.MethodsThe molecular mechanisms of fluoroquinolone resistance, including PMQR (plasmid-mediated quinolone resistance) as well as mutation in the QRDRs (quinolone-resistance determining regions) of gyrA, gyrB, parC and parE, among Enterobacterales isolated from 32 of 48 men undergoing a prostate biopsy between November 2015 and April 2016 were investigated. Before the TRUS-Bx procedure, all the patients received an oral antibiotic containing fluoroquinolones.ResultsIn total, 41 Enterobacterales isolates were obtained from rectal swabs. The MIC of ciprofloxacin and the presence of common PMQR determinants were investigated in all the isolates. Nine (21.9%) isolates carried PMQR with qnrS as the only PMQR agent detected. DNA sequencing of the QRDRs in 18 Enterobacterales (E. coli n = 17 and E. cloacae n = 1) isolates with ciprofloxacin MIC ≥ 0.25 mg/l were performed. Substitutions in the following codons were found: GyrA—83 [Ser → Leu, Phe] and 87 [Asp → Asn]; GyrB codon—605 [Met → Leu], ParC codons—80 [Ser → Ile, Arg] and 84 [Glu → Gly, Met, Val, Lys], ParE codons—458 [Ser → Ala], 461 [Glu → Ala] and 512 [Ala → Thr]. Six isolates with ciprofloxacin MIC ≥ 2 mg/l had at least one mutation in GyrA together with qnrS.ConclusionsThis study provides information on the common presence of PMQRs among Enterobacterales isolates with ciprofloxacin MIC ≥ 0.25 mg/l, obtained from men undergoing TRUS-Bx. This fact may partially explain why some men develop post-TRUS-Bx infections despite ciprofloxacin prophylaxis.

Mutational Diversity in the Quinolone Resistance-Determining Regions of Type-II Topoisomerases of Salmonella Serovars.

Quinolone resistance in bacterial pathogens has primarily been associated with mutations in the quinolone resistance-determining regions (QRDRs) of bacterial type-II topoisomerases, which are DNA gyrase and topoisomerase IV. Depending on the position and type of the mutation (s) in the QRDRs, bacteria either become partially or completely resistant to quinolone. QRDR mutations have been identified and characterized in Salmonella enterica isolates from around the globe, particularly during the last decade, and efforts have been made to understand the propensity of different serovars to carry such mutations. Because there is currently no thorough analysis of the available literature on QRDR mutations in different Salmonella serovars, this review aims to provide a comprehensive picture of the mutational diversity in QRDRs of Salmonella serovars, summarizing the literature related to both typhoidal and non-typhoidal Salmonella serovars with a special emphasis on recent findings. This review will also discuss plasmid-mediated quinolone-resistance determinants with respect to their additive or synergistic contributions with QRDR mutations in imparting elevated quinolone resistance. Finally, the review will assess the contribution of membrane transporter-mediated quinolone efflux to quinolone resistance in strains carrying QRDR mutations. This information should be helpful to guide the routine surveillance of foodborne Salmonella serovars, especially with respect to their spread across countries, as well as to improve laboratory diagnosis of quinolone-resistant Salmonella strains.

Antimicrobial resistance in Indian isolates of non typhoidal Salmonella of livestock, poultry and environmental origin from 1990 to 2017

A retrospective antimicrobial resistance study of nontyphoidal Salmonella enterica isolates from India during 1990–2017 was conducted to study the microbial susceptibility to antibiotics. A total of 271 Salmonella enterica isolates from poultry (n = 146), farm animals (n = 55) and environmental sources (n = 70) were tested for susceptibility using 15 antimicrobial drugs. The drug classes include aminoglycosides, phenicols, cephalosporins, penicillins, carbapenems, fluoroquinolones, and sulphonamide-trimethoprim. Study revealed that overall, 133 (49.08%) of 271 isolates were resistant to ≥ 1 antimicrobial drugs and 81 (29.89%) out of 271 isolates were multidrug resistant (resistance to ≥ 3 drugs). Majority (68.96%) of Typhimurium serovars (n = 87) were susceptible to all antibiotics tested, whereas only 5% Kentucky serovars (n = 40) were pan susceptible. All the drugs revealed decreasing trend of susceptibility from 1990 towards 2017 except cephalosporins and carbapenems. Statistical analysis of association between time period and antimicrobial resistance revealed a significance of < 0.05. Molecular detection of genetic determinants associated with antimicrobial resistance revealed the presence of genes like class I integrons, sul1, sul2, catIII, cmlA, dfrA, blaTEM, blaAmpC in the resistant isolates. Furthermore, plasmid mediated quinolone resistant determinants like qnrD and qnrS were also reported in the current study.

Carriage of Plasmid-Mediated Quinolone Resistance (PMQR) determinants by enrofloxacin-resistant ESBL- producing Escherichia coli recovered from abattoir leachate

ABSTRACT This study focused on the detection of PMQR genes in enrofloxacin-resistant isolates from a pool of extended spectrum β-lactamase ESBL- producing Escherichia coli recovered from abattoir leachate. Leachates were collected from three designated points and analysed using the streak plate technique. Isolates were identified, and subjected to antibiotic susceptibility testing. Enrofloxacin-resistant isolates were selected for the detection of PMQR genes. Eight isolates showing resistance to enrofloxacin were obtained. The resistance to antibiotics in the eight isolates was: cefotaxime (100%), oxytetracycline (87.5%), trimethoprim-sulfamethoxazole (87.5%) and imipenem (0%). The eight enrofloxacin-resistant bacteria carried both qnrA and qnrS, 62.5% carried qnrB, 25% and 37.5% respectively carried qepA and oqxAB efflux pumps, and 50% carried aac(6’)-lb-cr. There was co-carriage of PMQR determinants in the eight enrofloxacin-resistant isolates. Occurrence of antibiotic resistance and carriage of resistance genes in bacteria from abattoir leachate is a budding public health challenge, hence urgent action must be taken on this.

Isolation and Characterization of Shiga Toxin-Producing Escherichia coli from Retail Beef Samples from Eight Provinces in China.

While Shiga toxin-producing Escherichia coli (STEC) is a major foodborne pathogen worldwide, data on the molecular and phylogenetic properties of STEC isolates from retail beef samples in China remain scant. Fresh retail beef samples (n = 1062) were collected from eight provinces, and STEC isolates were recovered and characterized. PCR data showed that more than 50% of the samples were stx positive, and 82 STEC isolates were recovered from 14.8% (79/535) stx-positive enriched broths. In contrast, all ciprofloxacin resistant isolates (n = 19) and 13 cefotaxime (CTX) resistant isolates were eae positive and belonged to three serotypes: O111:H8, O26:H11, or O157:H7. Point mutations in quinolone resistance-determining regions and plasmid-mediated quinolone resistance determinants were identified in 16 and 20 isolates, respectively. BlaCTX-M and a point mutation (C-42T) in ampC promoter were detected in 15 and 8 of the CTX resistant isolates, respectively. In addition, macrolide resistance gene mphA was identified in eight azithromycin resistant O111:H8 isolates and one O26:H11 isolate. Single nucleotide polymorphism analysis demonstrated that the O26 and O157 isolates had multiple origins, but the O111 isolates were closely related. Taken together, our data demonstrated that several sequence types associated with hemolytic uremic syndrome from the retail beef samples in China had developed into dangerous multidrug resistant pathogens. The resistant phenotype can facilitate their transmission among the farm animals and human beings when there is an antimicrobial selective pressure.

Prevalence of the NTEKPC-I on IncF Plasmids Among Hypervirulent Klebsiella pneumoniae Isolates in Jiangxi Province, South China.

Infection caused by carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) has become a tricky health care threat in China and KPC-2 enzyme is a main factor mediating resistance to carbapenems of K. pneumoniae. Here, we report the characterization of the genetic environment of the blaKPC-2 gene in CR-hvKP clinical isolates from South China. Forty-five non-duplicated CR-hvKP isolates collected in Jiangxi Province from 2018 to 2019 were analyzed. Each of them were multidrug-resistant due to the presence not only of blaKPC-2 gene but also of other resistance determinants, including Metallo-β-lactamases (NDM-1), extended-spectrum β-lactamases (TEM-1, CTX-M-14, SHV-1), and plasmid-mediated quinolone resistance determinants (qnrS, aac(6′)-Ib-cr). After plasmid analyses of PCR-based replicon typing (PBRT), mapping PCR, amplicon sequencing, and whole-genome sequencing (WGS) were used to analyze the genetic environment of the blaKPC-2 gene. PCR analysis of pLVPK-like plasmids, Southern Blot, and mouse lethality assay were used to characterize the virulence phenotype of K. pneumoniae. Multilocus sequence typing (MLST) analysis showed ST11 CR-hvKP was the predominant clone. In conclusion, this is the first analysis of diverse genetic structures blaKPC-2 gene in CR-hvKP isolates from south China. Both the NTEKPC-I on the IncF plasmids and pLVPK-like virulence plasmids make contributions to the formation of CR-hvKP especially ST11 which need more attention.

Distribution of fluoroquinolone resistance determinants in Carbapenem-resistant Klebsiella pneumoniae clinical isolates associated with bloodstream infections in China

BackgroundThe rate of fluoroquinolone (FQ) resistance among carbapenem-resistant Klebsiella pneumoniae (CRKP) is high. The present study aimed to investigate the distribution of fluoroquinolone resistance determinants in clinical CRKP isolates associated with bloodstream infections (BSIs).ResultsA total of 149 BSI-associated clinical CRKP isolates collected from 11 Chinese teaching hospitals from 2015 to 2018 were investigated for the prevalence of fluoroquinolone resistance determinants, including plasmid-mediated quinolone resistance (PMQR) genes and spontaneous mutations in the quinolone resistance-determining regions (QRDRs) of the gyrA and parC genes. Among these 149 clinical CRKP isolates, 117 (78.5%) exhibited resistance to ciprofloxacin. The GyrA substitutions (Ser83 → IIe/Phe) and (Asp87 → Gly/Ala) were found among 112 (75.2%) of 149 isolates, while the substitution (Ser80 → IIe) of ParC was found in 111 (74.5%) of the 149 isolates. In total, 70.5% (105/149) of the CRKP isolates had at least two mutations within gyrA as well as a third mutation in parC. No mutations in the QRDRs were found in 31 ciprofloxacin susceptible CRKP isolates. Eighty-nine (56.9%) of 149 were found to carry PMQR genes including qnrS1 (43.0%), aac(6′)-Ib-cr (16.1%), qnrB4 (6.0%), qnrB2 (2.7%), and qnrB1 (1.3%). Nine isolates contained two or more PMQR genes, with one carrying four [aac(6′)-Ib-cr, qnr-S1, qnrB2, and qnrB4]. The co-existence rate of PMQR determinants and mutations in the QRDRs of gyrA and parC reached 68.5% (61/89). Seventy-four (83.1%, 74/89) PMQR-positive isolates harbored extended-spectrum beta-lactamase (ESBL)-encoding genes. Multilocus sequence typing (MLST) analysis demonstrated that the ST11 was the most prevalent STs in our study.ConclusionsMutations in the QRDRs of gyrA and parC were the key factors leading to the high prevalence of fluoroquinolone resistance among BSI-associated CRKP. The co-existence of PMQR genes and mutations in the QRDRs can increase the resistance level of CRKP to fluoroquinolones in clinical settings. ST11 CRKP isolates with identical QRDR substitution patterns were found throughout hospitals in China.

Prevalence of Plasmid-mediated quinolone resistance genes among Ciprofloxacin-nonsusceptible Escherichia coli and Klebsiella pneumoniae isolated from clinical isolates in Najaf, Iraq

Objectives: To investigate the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants in ciprofloxacin-Resistance Escherichia coli and Klebsiella pneumoniae isolated from patients in Najaf. Methods: A total of 134 non-duplicate isolates of ciprofloxacin-resistant E. coli (n=60) and K. pneumoniae (n=74) from clinical isolates cultures were obtained. The qnrA, qnrB, qnrS, qnrC, qnrD, aac(6')-Ib-cr, and qepA genes were detected using polymerase chain reaction (PCR). Results: The study was revealed out of 134 non-repetitive quinolones resistant E. coli and K. pneumoniae isolates, 46(76.6%) E. coli and 47 (66.7%) K. pneumoniae isolates had at least one PMQR quinolones resistant gene. 22 (16.4%) qnrB-positive isolates were identified, including 5 (8.33%) E. coli, and 17(23%) K. pneumoniae, 8(5.97%) qnrS were identified, including 7(11.7%) E. coli, and 1(1.4%) K. pneumoniae, 3(2.2%) qnr A were identified, including 1(1.6%) E. coli, and 22(2.7%) K. pneumoniae, 3 qepA -positive isolates were identified, including 3(4.1%) K. pneumoniae. The aac (6')-Ib-cr variant gene was detected in 85(63.4%) including 46(79.7%) E. Coli and 39 (52.7%) of the clinical isolates K. Pneumoniae. Conclusion: This study detected several of plasmid-mediated quinolone resistance genes among ciprofloxacin-nonsusceptible E. coli and K. pneumoniae isolated, many of them detected for the first time in Najaf, Iraq, like qnrA, qnrB, qnrS genes with predominance of qnrB gene, in addition to qepA gene in K. pneumoniae.

Characterization of Multidrug-Resistant Biofilm-Producing Escherichia coli and Klebsiella pneumoniae in Healthy Cattle and Cattle with Diarrhea

This study describes comparative occurrence and characterization of multidrug-resistant (MDR) Escherichia coli and Klebsiella pneumoniae (KP) in healthy cattle (HC) and cattle with diarrhea (DC) in India. During 2018-2020, 72 MDR isolates, including 35 E. coli (DC: 27; HC 8) and 37 K. pneumoniae (DC: 34; HC: 3), from 251 rectal swabs (DC: 219; HC: 32) were investigated for extended-spectrum beta-lactamase (ESBL), AmpC type β-lactamase and carbapenemase production, antimicrobial susceptibility profile, biofilm production, and efflux pump activity. Fifty-five MDR isolates were ESBL producers (ESBLPs) (DC: 50; HC: 5) and ESBLPs from DC were coresistant to multiple antibiotics. The blaCTX-M gene (50) was the most frequently detected β-lactamases followed by blaAmpC (22), blaTEM1 (13), blaCMY-6 (6), blaOXA1 (5), blaPER (2), blaDHA, and blaFOX and blaSHV12 (1 each). Plasmid-mediated quinolone resistance determinants qnrB, qnrS, qnrA, and qepA were detected in 18, 16, 2, and 3 isolates, respectively. Twenty three isolates revealed mutation in gyrA and parC genes. Tetracycline-resistance markers tetA, tetB, tetC, and tetE were detected in 33, 10, 3, and 2 isolates, respectively. Only one of the 41 imipenem-resistant isolates harbored blaNDM-5 and two were colistin-resistant. Altogether, 20 MDR isolates were strong biofilm producers and 19 harbored different virulence factors. This is the first ever report from India on the presence of MDR Enterobacteriaceae with resistance to even last-resort antimicrobials in the bovine diarrhea.

Predominance and clonal spread of CTX-M-15 in cefotaxime-resistant Klebsiella pneumoniae in Korea and their association with plasmid-mediated quinolone resistance determinants

Introductionβ-lactams and fluoroquinolones are extensively used worldwide in the treatment of infections caused by Enterobacterales. In this study, we investigated the prevalence of extended-spectrum β-lactamases (ESBL), their correlation with plasmid-mediated quinolone resistance determinants (PMQR) and clonal distribution among the cefotaxime-resistant K. pneumoniae isolates. MethodsIn Korea, a total of 429 K. pneumoniae collected in 2015 were studied. Antimicrobial susceptibility test for cefotaxime, ciprofloxacin and levofloxacin was performed by broth microdilution method. By PCR and/or sequencing, mutations in gyrA and parC genes, PMQR genes and ESBL were identified. Multilocus-sequence-type (MLST) was determined for isolates harboring CTX-M-15. ResultsAmong the 149 K. pneumoniae showing cefotaxime MICs of >1 μg/ml, 142 (95.3%) isolates were ESBL-producers and CTX-M-15 was predominant (99 isolates). Among the 142 ESBL-producers, mutations in gyrA and parC were found in 112 (78.9%) and 93 isolates (65.5%), respectively. PMQR genes were detected in 141 isolates and the non-susceptibility rate to ciprofloxacin and levofloxacin was 95.1% (135/142) and 82.4% (117/142), respectively. The most frequently found PMQR combination was qnrB-aac(6′)-Ib-cr-oqxAB, (58/142, 40.8%). By MLST, four major STs/CC: ST48, ST392, ST307 and CC15 accounted for 67% of the CTX-M-15 producers and the prevalence of qnrB was significantly higher in these four major STs/CC than other groups (P = 0.004). Of note, we found the additive effect of PMQR genes; the more PMQR genes, the higher ciprofloxacin MICs. ConclusionsCTX-M-15 was predominant among the cefotaxime-resistant K. pneumoniae and co-harboring CTX-M-15 and PMQR genes, especially qnrB, seems to contribute the spread of high risk clones.

Quinolone resistant Salmonella species isolated from pediatric patients with diarrhea in central Iran

BackgroundThis study aimed to investigate the frequency and the antibiotic resistance patterns of Salmonella species that were isolated from infectious diarrhea samples taken from pediatric patients in central Iran.MethodsThe study analyzed 230 stool specimens that were cultured on XLD, MacConkey agar and GN broth. Polymerase chain reaction (PCR) assay was used to identify the Salmonella genus. The antibiotic resistance profiles and the frequency of quinolone and integron genes were obtained.ResultsOut of 230 samples of infectious diarrhea, 21 (9.1%) cases of Salmonella spp. were identified using culture methods. Another 28 (12.1%) samples had positive PCR results, with S. serovar Paratyphi B and C (9/21; 42.8%) and S. Typhi (3/21; 14.3%) being the most recognized. The highest antibiotic resistance rates were found for nalidixic acid (15/21; 71.4%), tetracycline (9/21; 42.8%). However, six (28.5%) of isolates were found resistant to cotrimoxazole, ampicillin and chloramphenicol. Among the plasmid-mediated quinolone resistance (PMQR) determinants, qnrS, qnrA, and qnrB were positive in (9/15; 60%), (6/15; 40%) and (3/15; 20%) of the isolates, respectively. Class 1 and 2 integrons were identified in 15 (71.4%) and 3 (14.3%) isolates, respectively.ConclusionHigh rates of quinolone resistant and low frequency of MDR Salmonella spp. isolates were identified in central Iran, similar to findings in other parts of Asia. To prevent the spread of these resistant strains, the antimicrobial resistance of Salmonella spp. isolates should be under constant surveillance, and empiric antibiotic therapy should be adapted appropriately.

Epidemic and molecular characterization of fluoroquinolone-resistant Shigella dysenteriae 1 isolates from calves with diarrhea

BackgroundThe widespread distribution of antimicrobial-resistant Shigella has become a recurrent challenge in many parts of the developing world. Previous studies indicate that the host of Shigella has expanded from humans to animals. This study aimed to investigate the prevalence of fluoroquinolone resistance and associated molecular characterization of S. dysenteriae 1 isolated from calves.ResultsAll 38 unduplicated S. dysenteriae 1 isolates were collected from calves in Gansu Province from October 2014 to December 2016. According to MLST and PFGE analysis, these isolates were separated into 4 and 28 genotypes, respectively. The most common STs identified were ST228 (34.21%, 13/38) and ST229 (39.47%, 15/38), which were first found in the present study. All isolates harbored virulence genes, and the incidence of the seven virulence genes were ipaH (100%), ipaBCD (92.11%), stx (73.68%), ial (57.89%), sen (28.95%), set1A and set1B (0%). According to the results of antimicrobial susceptibilities, 76.32% (29/38) were resistant to fluoroquinolone and showed multidrug resistance. In a study on the polymorphism of quinolone resistance–determining region (QRDR) of gyrA/B and parC/E genes, we identified two mutations in gyrA (Ser83 → Leu and Asp87 → Asn) and parC (Ser80 → Ile and Ser83 → Leu), respectively. Among them, 55.17% (16/29) of resistant strains had the gyrA point mutations (Ser83 → Leu) and parC point mutation (Ser83 → Leu). Moreover, 41.38% (12/29) of isolates had all five point mutations of gyrA and parC. In addition, the prevalence of the plasmid-mediated quinolone resistance (PMQR) determinant genes was also investigated. All 29 fluoroquinolone-resistant isolates were positive for the aac (6′)-Ib-cr gene but negative for qepA, except for SD001. In addition, only 6 (20.69%, 6/29) isolates harbored the qnr gene, including two with qnrB (6.90%, 2/29) and four with qnrS (13.79%, 4/29).ConclusionGiven the increased common emergence of multidrug resistant isolates, uninterrupted surveillance will be necessary to understand the actual epidemic burden and control this infection.

1667. Change in characteristics of community-onset ciprofloxacin-resistant E. coli isolates causing community-acquired acute pyelonephritis in South Korea

BackgroundThe aim of this study was to examine the change in characteristics of community-onset ciprofloxacin-resistant (CIP-R) E. coli isolates causing community-acquired acute pyelonephritis (CA-APN) in South Korea between 2010-2011 and 2017-2018.Methods E. coli samples isolated from the blood or urine were collected from patients with CA-APN aged 19 years and more who were admitted to 8 Korean hospitals from September 2017 to August 2018, prospectively. One isolate was collected from each patient. Phylogenetic typing, multilocus sequence typing (MLST), and molecular characterization of β-lactamase resistance and plasmid-mediated quinolone resistance (PMQR) determinants were performed. The data were compared with those from the previous study with same design in 2010-2011.ResultsA total of 346 and 300 isolates were collected during 2017-2018 and 2010-2011, respectively. Among them, 76 (22.0%) and 77 (25.7%) were CIP-R isolates. Significantly higher antimicrobial resistance against ampicillin (75.7% vs. 100%, P < 0.001) and cefotaxime (23.9% vs. 77.9%, P < 0.001) were observed for isolates in 2017-2018 compared to those in 2010-2011. The proportion of phylogenic group B2 had increased significantly (44.7% vs. 79.2%, P < 0.001). As for MLST, the proportion of ST131 (27.6% vs. 66.2%, P < 0.001) had increased while that of ST393 (18.4% vs. 3.9%, P =0.004) had decreased significantly. Higher proportion of CIP-R E. coli isolates in 2017-2018 had extended-spectrum β-lactamase (ESBL)/plasmid-mediated AmpC β-lactamase (PABL) (23.7% vs. 79.2%, P < 0.001) and PMQR determinant (11.8% vs. 40.8%, P < 0.001) compared to those in 2010-2011.Phlogenetic tree Analyzed by SplitsTree ConclusionAmong uropathogenic CIP-R E. coli isolates in South Korea, ST131 predominance had become more prominent and the proportion of containing ESBL/PABL and/or PMQR determinants had increased.Disclosures All Authors: No reported disclosures

Fluoroquinolone Resistance in Clinical Isolates of Klebsiella Pneumoniae

Introduction Fluoroquinolones are widely used broad-spectrum antibiotics. Recently, increased rate of resistance to this antibiotic has been observed in Klebsiella pneumoniae. The aim of the present study was to determine the presence of quinolone resistance determining regions (QRDR) mutation genes and plasmid-mediated quinolone resistance (PMQR) determinants in clinical isolates of ciprofloxacin-resistant K. pneumoniae. Material and Methods The study included 110 nonduplicate ciprofloxacin-resistant K. pneumoniae clinical isolates. Antibiotic susceptibility testing by disk diffusion method and minimum inhibitory concentration (MIC) by agar dilution methods for ciprofloxacin was performed according to the recommendations of Clinical Laboratory Standards Institute. The presence of QRDR genes and PMQR genes was screened by polymerase chain reaction (PCR) amplification. Result All 110 isolates were resistance to ciprofloxacin, levofloxacin, and ofloxacin. As much as 88% of the isolates exhibited high-level of MIC to ciprofloxacin. Among the 110 isolates, 94(85%) harbored gyrA and 85 (77%) gyrB. The parC and parE genes were detected in 88 (80%) and 64 (58%) isolates. qnrB was detected in 13 (12%) isolates and qnrS in 5 (4.5%) isolates. Two (1.8%) isolates carried both qnrB and qnrS genes. The acc (6')-Ib-cr gene was found in 98 (89%) isolates and oqxAB was detected in 7 (6.3%) isolates. One (0.9%) isolate carried qnrB, acc(6')-Ib-cr and oqxAB genes. Conclusion The prevalence of acc (6')-Ib-cr gene is high among PMQR determinants, followed by qnrB, oqxAB and qnrS.

First Detection of the Plasmid-Mediated Quinolone Resistance Determinants qnrB, qnrS and aac(6’)-Ib-cr in Extended Spectrum Beta-Lactamases-producing Klebsiella pneumonia in Bouaké, Côte d’Ivoire

Aims: The aims of the present study were to investigate the presence of Plasmid-Mediated Quinolone Resistance (PMQR) determinants and the association of these determinants with Extended Spectrum Beta-Lactamases (ESBLs) genes in ESBL-producing Klebsiella pneumoniae isolates from Teaching Hospital of Bouaké, Côte d’Ivoire.&#x0D; Study Design: It is a retrospective study.&#x0D; Place of Study: Bacteriology-Virology Laboratory of Teaching Hospital, Bouaké, Côte d'Ivoire.&#x0D; Methodology: From January 2015 to December 2016, 96 ESBL-producing Klebsiella pneumoniae isolates were collected from several specimens. Antimicrobial susceptibility of isolates was tested using the standard disk-diffusion method on Mueller-Hinton and interpretation according to recommendations of the 2017 EUCAST. These isolates analyzed for the detection of ESBL (blaCTX-M, blaTEM and blaSHV) and PMQR genes (aac(6’)-Ib-cr, qnrB and qnrS) using simplex PCR.&#x0D; Results: Of the 96 ESBL-producing strains, 85 (88.55%) harbored at least one of the ESBL genes tested. Out of the 85 strains encoding ESBL genes, 96.47% carried blaCTX-M and 92.94% blaSHV and blaTEM genes. Eighty nine (89.6%) of the 96 ESBL producing-isolates were resistant to ciprofloxacin and 84.4% to norfloxacin. Among the 96 strains, 80 (83.33%) were found harboring at least one PMQR gene consisting of 78 (81.3%) aac(6’)-Ib-cr, 61 (63.5%) qnrB and 15 (15.6%) qnrS. Among the PMQR-positive strains, 68.4% coharbored qnrB+acc(6’)-Ib-cr genes, 10.5% qnrB+qnrS+acc(6’)-Ib-cr and 6.6% qnrS+acc(6’)-Ib-cr. The qnrB gene was always linked to aac(6’)-Ib-cr gene. Aac(6’)-Ib-cr gene showed the highest association with three ESBL genes (87.6%), followed by qnrB gene (70.6%), then qnrS (17.7%).&#x0D; Conclusion: The PMQR genes were highly prevalent in ESBL-producing Klebsiella pneumoniae, primarily the aac(6’)-Ib-cr gene. The high associated was observed between ESBL and PMQR genes, notably with the aac(6’)-Ib-cr gene.

Prevalence and Characterization of Plasmid-Mediated Quinolone Resistance Determinants qnr and aac(6')-Ib-cr in Ciprofloxacin-Resistant Escherichia coli Isolates from Commercial Layer in Korea.

The prevalence and characterization of plasmid-mediated quinolone resistance (PMQR) determinants in ciprofloxacin-resistant Escherichia coli isolated from a Korean commercial layer farm were studied. A total of 45 ciprofloxacin-resistant E. coli isolates were recovered and all isolates were multidrug-resistant. Eight isolates have the PMQR genes aac(6')-Ib-cr, qnrS1, and qnrB4, and seven isolates exhibited double amino acid exchange at both gyrA and parC, and have high fluoroquinolone minimum inhibitory concentrations. Five transconjugants demonstrated transferability of PMQR and β-lactamase genes and similar antimicrobial resistance. Because PMQR genes in isolates from commercial layer chickens could enter the food supply and directly affect humans, control of ciprofloxacin resistance is needed.