Prevalence and characterization of plasmid-mediated quinolone resistance genes in Salmonella isolated from poultry in Korea. - Related Documents




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205501.0000Prevalence and characterization of plasmid-mediated quinolone resistance genes in Salmonella isolated from poultry in Korea. The purpose of this study was to investigate the prevalence and characteristics of plasmid-mediated quinolone resistance (PMQR) genes qnr, aac(6')-Ib-cr, and qepA in a total of 185 non-duplicate Salmonella spp. isolated from hatcheries, poultry farms, and poultry slaughterhouses during the period 2001 to 2010 in Korea. Additionally, mutation analysis of quinolone resistance determining regions (QRDRs), conjugation experiments, and plasmid analysis were performed in the PMQR-positive isolates. Among the 185 isolates, six (3.2%) contained qnr genes (two qnrB4 and four qnrS1) but none carried the aac(6')-Ib-cr or qepA genes. Among the six PMQR-positive isolates, one showed a single mutation (Ser83-Phe substitution) in the QRDRs of gyrA. Among them, three were non-susceptible (intermediate or resistant) to nalidixic acid (minimum inhibitory concentration [MIC] ≥256 µg/ml), ciprofloxacin (MIC 2 µg/ml), and levofloxacin (MIC 4 µg/ml), but others were susceptible to all of the three fluoroquinolones. They were resistant to six or more antimicrobial agents tested and were able to transfer quinolone resistance to recipient Escherichia coli J53 by conjugation. By performing a hybridization test, plasmids harbouring qnrB4 and qnrS1 genes were less than 8 kb and about 70 kb in size, respectively. The horizontal dissemination of qnrS1 gene was mediated by IncN plasmid. Compared with the recipient strain, MICs of the transconjugants increased two-fold to four-fold for nalidixic acid, and eight-fold to 16-fold for ciprofloxacin and levofloxacin. This report is the first to describe the detection of qnr genes in Salmonella spp. isolated from poultry in Korea. Widespread horizontal transfer of these genes among bacteria may be a serious public health concern because these can rapidly increase fluoroquinolone resistance. To ensure the public health, it is essential to continuously survey and carefully monitor the spread of PMQR genes in Salmonella from poultry.201323607509
205410.9999A survey of plasmid-mediated fluoroquinolone resistance genes from Escherichia coli isolates and their dissemination in Shandong, China. Bacterial resistance to fluoroquinolones result from mutations in the quinolone resistance-determining regions of the drug targets, overexpression of efflux pumps, and/or the more recently identified plasmid-mediated low-level resistance mechanisms. We investigated the prevalence of and characterized plasmid-mediated fluoroquinolone resistance genes (qnrA, qnrB, qnrS, aac(6')-Ib-cr, and qepA) by polymerase chain reaction in fluoroquinolone-resistant Escherichia coli (n = 530) isolated from a chicken farm, a pig farm, and hospitalized patients in Shandong, China, in 2007. The aac(6')-Ib-cr gene was the most prevalent resistance gene that was detected in bacteria isolated from all sources. Next was the qnrS gene, which was predominantly present in isolates from the pig farm. Only eight (5.8%) isolates from hospital patients were found to possess the qepA gene, and these isolates were first reported in qepA-carrying E. coli from humans in China. The qnrA and qnrB genes were not detected in any of the isolates. Further, most of the isolates were also resistant to beta-lactams and aminoglycosides as determined by the broth microdilution method. Pulsed-field gel electrophoresis analysis of the E. coli isolates with similar resistance patterns that also carried resistance genes showed great genomic diversity among these bacteria, suggesting that the multiresistant E. coli isolates carrying the qnr, aac(6')-Ib-cr, or qepA genes were not derived from a specific clone, but represented a wide variety of different genotypes. The results of Southern hybridization revealed that qepA, qnrS, and parts of aac(6')-Ib-cr genes were localized on plasmids and/or chromosome. qepA and aac(6')-Ib-cr genes were colocalized with aac(6')-Ib-cr and qnrS genes, respectively, on the same plasmids. Our study demonstrated that two different genes (qepA and aac(6')-Ib-cr) were identified on the same plasmid in E. coli strains derived from patients and qnrS and aac(6')-lb-cr genes on the same plasmid in an E. coli strain of animal origin.201019911944
205220.9999Plasmid-mediated quinolone resistance in Escherichia coli isolates from commercial broiler chickens and selection of fluoroquinolone-resistant mutants. Plasmid-mediated quinolone resistance (PMQR) is a potential concern for animal husbandry and public health. Escherichia coli isolates from a total of 109 fecal samples collected from 6 commercial broiler farms between 2007 and 2011 were examined for PMQR genes, and transfer of these genes was tested by conjugation analysis to elucidate the prevalence and spread of PMQR in broiler chickens. Two isolates from 2 farms harbored the aac(6')-Ib-cr gene that was not detected in plasmids using Southern blot analysis of S1 nuclease-digested genomic DNA separated by pulsed-field gel electrophoresis. In these 2 isolates, nucleotide mutations in the gyrA and parC genes that result in amino acid substitutions were detected. Additionally, a total of 6 isolates originating from 6 chickens from the 2 farms were positive for the qnrS1 gene. In 2 of the 6 isolates, the qnrS1 gene was transferred to a recipient strain. Two transconjugants harboring the qnrS1 gene were cultured on media supplemented with successively higher concentrations of enrofloxacin (ERFX). After a 5-time subcultivation, the ERFX MICs reached 8 and 16 μg/mL, and no nucleotide mutations were detected in the gyrA, gyrB, parC, and parE genes. Our results suggest that the prevalence of PMQR was relatively low in broiler chickens and that exposure of bacteria carrying PMQR genes to the selective pressure of fluoroquinolones can result in resistance to fluoroquinolone, which is not caused by mutations in genes encoding topoisomerases.201931198966
205330.9999Replicon typing of plasmids in environmental Achromobacter sp. producing quinolone-resistant determinants. This study aimed to investigate the antimicrobial resistance profile to quinolones, the presence of quinolone-resistant determinants and the plasmid replicon typing in environmental Achromobacter sp. isolated from Brazil. Soil and water samples were used for bacterial isolation. The antimicrobial susceptibility testing was performed by minimum inhibitory concentration method. The detection of mutations in the quinolone resistance-determining regions (QRDR) genes, the presence of plasmid-mediated quinolone resistance (PMQR) genes, and plasmid replicons were performed by PCR. A total of 16 isolates was obtained from different cultures, cities, and states of Brazil. All isolates were non-susceptible to ciprofloxacin, norfloxacin, and levofloxacin. Some mutations in QRDR genes were found, including Gln-83-Leu and Asp-87-Asn in the gyrA and Gln-80-Ile and Asp-84-Ala in the parC. Different PMQR genes were detected, such as qnrA, qnrB, qnrS, oqxA, and oqxB. Three different plasmid families were detected, being most presented the ColE-like, followed by IncFIB and IncA/C. The presence of different PMQR genes and plasmids in the isolates of the present study shows that environmental bacteria can act as reservoir of important genes of resistance to fluoroquinolones, which is of great concern, due to the potential of horizontal dissemination of these genes. Besides that, there are no studies reporting these results in Achromobacter sp. isolates.201830357960
202440.9998Research Note: Longitudinal monitoring of chicken houses in a commercial layer farm for antimicrobial resistance in Escherichia coli with special reference to plasmid-mediated quinolone resistance. Plasmid-mediated quinolone resistance (PMQR) genes located on conjugative plasmids can be transferred to other bacteria in the absence of antimicrobial selective pressure. To elucidate the prevalence of resistance, including PMQR in an egg-producing commercial layer farm in western Japan where no antimicrobials were used, minimum inhibitory concentrations (MIC) for a total of 375 Escherichia coli isolates obtained from chicken houses in the farm between 2012 and 2017 were determined using the agar dilution methods. Eighty-seven isolates resistant to oxytetracycline (OTC) accounted for 23.0% of the tested isolates, followed by isolates resistant to dihydrostreptomycin (DSM) (18.4%), sulfisoxazole (18.1%), ampicillin (AMP) (14.4%), trimethoprim (TMP) (14.4%), and nalidixic acid (10.1%). The prevalence rate of multidrug-resistant (MDR) isolates-which are resistant to 3 or more antimicrobial classes, including β-lactams, aminoglycosides, quinolones, folate pathway inhibitors, tetracyclines, and phenicols-was inversely related to the age of chickens at the time of bacterial examination. Probably, the prevalence of MDR isolates in layer chickens may have decreased with age owing to the absence of selective pressure. Furthermore, 45 isolates exhibiting enrofloxacin MICs of more than 0.25 μg/mL were examined for PMQR genes. The transfer of PMQR genes was tested by conjugation analysis. Southern blot analysis of genomic DNA revealed that the qnrS1 (5 isolates), qnrS2 (1 isolate), and qnrS13 genes (1 isolate) were located on plasmids with sizes ranging from approximately 60 to 120 kpb. In 1 of the 5 qnrS1-positive isolates and in an isolate with qnrS13, the qnrS genes were transferred to recipient strains. The plasmid harboring the qnrS1 gene was typed as IncF by PCR-based replicon typing. On this plasmid, the bla(TEM), aadA, tetA, and dfrA1 genes responsible for resistance to AMP, DSM, OTC, and TMP, respectively, were detected. The tetA gene was detected in the plasmid harboring the qnrS13 gene, which was typed as IncI1. These results suggest that despite the low prevalence of quinolone resistance in this farm, various PMQR genes, located on diverse plasmids, exist.202032036966
96450.9998Distribution of plasmid-mediated quinolone resistance in Gram-negative bacteria from a tertiary hospital in Nigeria. BACKGROUND: Until recently, mechanisms of resistance to quinolones in Gram-negative bacteria were believed to be only chromosome encoded. However, emergence of plasmid-mediated quinolone resistance (PMQR) has been reported worldwide. AIM: This study investigated distribution of PMQR in Gram-negative bacteria from a tertiary hospital in eastern part of Nigeria. MATERIALS AND METHODS: Seventy-one nonduplicate Gram-negative bacterial isolates of eight species were analyzed for antimicrobial susceptibility, genotypic detection of various PMQRs, typed by random amplified polymorphic DNA (RAPD) and analysis of plasmids present, including replicon typing. RESULTS: The minimum inhibitory concentrations showed MIC90values as high as 256 μg/ml for fluoroquinolones. Carriage of PMQR was found to be 35.2%. Twenty (28.2%) isolates carried various qnr genes, of which seven (9.9%) qnrA1; four (5.6%) qnrB1; eight (11.3%) qnrS1 while one (1.4%) encoded qnrD1. Eighteen (25.4%) isolates were positive for aac(6')-Ib-cr while carriage of multiple genes exists in some strains. Similarly, 13 isolates (18.7%) were found to carry PMQR efflux pump gene, qepA. Conjugation experiments revealed that the plasmids once transferred coded for fluoroquinolone resistance. The transconjugant strains carried a common plasmid estimated to be 65 kb. These plasmids were untypable for replicon/incompatibility. Typing revealed high diversity among all species tested with no identical RAPD pattern seen. CONCLUSION: This study further confirms high level resistance to many antimicrobials in different species of Gram-negative bacteria including fluoroquinolones and spread of PMQR genes in Southern Nigeria.201627510669
96860.9998Molecular analysis of antimicrobial resistance in gram-negative bacteria isolated from fish farms in Egypt. As little is known about antimicrobial resistance genes in fish farms, this study was conducted to monitor the incidence and prevalence of a wide range of antimicrobial resistance genes in Gram-negative bacteria isolated from water samples taken from fish farms in the northern part of Egypt. Ninety-one out of two hundred seventy-four (33.2%) non-repetitive isolates of Gram-negative bacteria showed multidrug resistance phenotypes and harbored at least one antimicrobial resistance gene. PCR and DNA sequencing results showed that 72 (26.3%) isolates contain tetracycline resistance genes and 19 (6.9%) isolates were positive for class 1 integrons with 12 different gene cassettes. The beta-lactamase-encoding genes were identified in 14 (5.1%) isolates. The plasmid-mediated quinolone resistance genes, qnr and aac(6')-Ib-cr, were identified in 16 (5.8%) and 3 (1.1%) isolates, respectively. Finally, the florphenicol resistance gene, floR, was identified in four (1.5%) isolates. To the best of our knowledge, this is the first report for molecular characterization of antimicrobial resistance in Gram-negative bacteria isolated from fish farms in Africa.201020145377
96570.9998Molecular Characterization of Multidrug-Resistant Escherichia coli Isolates from Bovine Clinical Mastitis and Pigs in the Vojvodina Province, Serbia. The aim of the study was to characterize multidrug-resistant (MDR) Escherichia coli isolates collected in Serbia from bovine clinical mastitis cases and diseased pigs, mainly with molecular methods. A total of 48 E. coli isolates was collected during the years 2013-2014, of which 22 were MDR and were included in further analysis. Phylogenetic typing showed that 17 isolates belonged to group A, while two isolates were classified in group B1 and a single one in group D. All isolates showed unique macrorestriction patterns. Phenotypic susceptibility testing revealed resistances of the isolates against up to 13 antimicrobial agents, including resistance to fluoroquinolones. A wide variety of resistance genes was detected by PCR amplification and sequencing of amplicons. Sequence analysis of the quinolone resistance determining regions of topoisomerase genes revealed mutations in gyrA, parC, and/or parE. Plasmid-mediated quinolone resistance genes were detected in two porcine (aac-6'-Ib-cr and qnrS, respectively) isolates and a single bovine (aac-6'-Ib-cr) isolate. Resistance genes were found to be located on conjugative plasmids in 16 cases, many of which conferred a multidrug resistance phenotype. In conclusion, the plentitude of resistance genes located on conjugative plasmids and integrons in E. coli from cows and pigs in Vojvodina, Serbia, pose a high risk for horizontal gene transfer in bacteria from livestock husbandry.201828520501
173480.9998Identification and characterization of plasmid-mediated quinolone resistance determinants in Enterobacteriaceae isolated from healthy poultry in Brazil. The expression of plasmid-mediated quinolone resistance (PMQR) genes confers low-level quinolone and fluoroquinolones resistance alone. However, the association to chromosomal resistance mechanisms determines an expressively higher resistance in Enterobacteriaceae. These mechanisms are horizontally disseminated within plasmids and have contributed to the emergence of bacteria with reduced susceptibility or resistant to therapies worldwide. The epidemiological characterization of PMQR dissemination is highly relevant in the scientific and medical context, to investigate the dissemination within enterobacteria, from different populations, including humans and food-producing animals. In the present study, 200 Enterobacteriaceae isolates were harvested from poultry with cloacal swabs and identified as Escherichia coli (90.5%), Escherichia fergusonii (5.5%), Klebsiella oxytoca (2.5%) and Klebsiella pneumoniae (1.5%). Among isolates evaluated, 46 (23%) harboured PMQR genes including qnrB (43/200), qnrS (2/200) and aac(6')-Ib-cr (1/200). All isolates carrying PMQR genes showed multidrug-resistance phenotype. The 36 E. coli isolates showed 18 different PFGE types. All E. fergusonii isolates showed the same PFGE type. The two Klebsiella oxytoca belonged to two different PFGE types. The phylogenetic groups A, B1, and D were found among the E. coli harboring PMQR genes. Based on the phylogenetic analysis and PFGE, the population structure of E. coli isolates was diverse, even within the same farm. All isolates carrying qnrB and qnrS genes also harboured ColE-like plasmids. The Southern blot hybridization using the S1-PFGE revealed that the qnrB genes were located on low molecular weight plasmids, smaller than 10Kb. Resistance plasmids were sequenced and showed 100% identity with plasmid pPAB19-3. The association of PMQR genes with mobile genetic elements, such as transferable plasmids, favours the selection and dissemination of (fluoro) quinolones resistant bacteria among food-producing animals, and may play an important role in the current increased prevalence of resistant bacteria in different environments reported worldwide.201829427764
290790.9998Prevalence of tetracycline resistance genes and identification of tet(M) in clinical isolates of Escherichia coli from sick ducks in China. Tetracycline resistance is one of the most frequently encountered resistance properties in bacteria of animal origin. The aim of the present study was to investigate the prevalence and diversity of tetracycline resistance (tet) genes among Escherichia coli clinical isolates from diseased ducks in China and to report the identification and sequencing of the tet(M) gene. The susceptibility of 85 Escherichia coli strains to tetracyclines was determined by broth microdilution, and the presence of tet genes was investigated by multiplex PCR. All of the 85 isolates were fully resistant to both oxytetracycline and tetracycline, and 76.5 % were resistant to doxycycline. Seventy-seven of the isolates (90.6 %) encoded multiple tet genes, with 17.6, 38.8 and 34.1 % encoding two, three and four tet genes, respectively, and only 7.1 % encoded a single tet(A) gene. The MICs of oxytetracycline and tetracycline for all isolates ranged from 16 to ≥128 µg ml(-1) with a MIC90 of >128 µg ml(-1), regardless of the type or number of tet genes encoded. Isolates containing tet(M) commonly had more than one tet gene per strain. The doxycycline resistance rate in the tet(M)-positive isolates was significantly higher than in the tet(M)-negative isolates (P<0.05). A full-length tet(M) gene, including the promoter region, was obtained by PCR in seven of the 41 tet(M)-positive isolates and was sequenced and cloned. The cloned tet(M) gene conferred resistance to tetracyclines in the recombinant Escherichia coli host strain. These results revealed that, in these isolates, the prevalence of multiple tet genes was strikingly high and that tet(M) played a role in doxycycline resistance.201323475906
2056100.9998Mechanisms of resistance in nontyphoidal Salmonella enterica strains exhibiting a nonclassical quinolone resistance phenotype. Nontyphoidal Salmonella enterica strains with a nonclassical quinolone resistance phenotype were isolated from patients returning from Thailand or Malaysia to Finland. A total of 10 isolates of seven serovars were studied in detail, all of which had reduced susceptibility (MIC > or = 0.125 microg/ml) to ciprofloxacin but were either susceptible or showed only low-level resistance (MIC < or = 32 microg/ml) to nalidixic acid. Phenotypic characterization included susceptibility testing by the agar dilution method and investigation of efflux activity. Genotypic characterization included the screening of mutations in the quinolone resistance-determining regions (QRDR) of gyrA, gyrB, parC, and parE by PCR and denaturing high-pressure liquid chromatography and the amplification of plasmid-mediated quinolone resistance (PMQR) genes qnrA, qnrB, qnrS, qnrD, aac(6')-Ib-cr, and qepA by PCR. PMQR was confirmed by plasmid analysis, Southern hybridization, and plasmid transfer. No mutations in the QRDRs of gyrA, gyrB, parC, or parE were detected with the exception of a Thr57-Ser substitution within ParC seen in all but the S. enterica serovar Typhimurium strains. The qnrA and qnrS genes were the only PMQR determinants detected. Plasmids carrying qnr alleles were transferable in vitro, and the resistance phenotype was reproducible in Escherichia coli DH5alpha transformants. These data demonstrate the emergence of a highly mobile qnr genotype that, in the absence of mutation within topoisomerase genes, confers the nontypical quinolone resistance phenotype in S. enterica isolates. The qnr resistance mechanism enables bacteria to survive elevated quinolone concentrations, and therefore, strains carrying qnr alleles may be able to expand during fluoroquinolone treatment. This is of concern since nonclassical quinolone resistance is plasmid mediated and therefore mobilizable.200919596880
1181110.9998Plasmid-mediated quinolone resistance genes transfer among enteric bacteria isolated from human and animal sources. This research investigates the transferability of plasmid-mediated quinolone resistance (PMQR) genes among enteric bacteria isolates in human and animal samples, as well as its implication on resistance of recipient cells. A total of 1,964 strains of five different enteric bacteria species (Escherichia coli, Salmonella sp., Shigella sp., Klebsiella sp. and Aeromonas sp.) were screened for plasmid-mediated quinolone resistance (PMQR) genes from a population of quinolone resistant (Q-r) isolates. Screening for PMQR isolates was achieved by plasmid curing using sub-lethal concentration of Sodium Dodecyl Sulphate and PMQR genes (qnrA, qnrB, qnrS, Aac(6')-Ib-crand Qep A) were detected by polymerase chain reaction (PCR). Conjugation and transformation experiments were attempted to ascertain transfer of genes from the Q-r isolates to a susceptible, standard recipient, E. coli J53-2. The minimum inhibitory concentration (MIC) was determined before and after gene transfer, using E-test strips. Results indicate that percentage resistance to the quinolones (Qs): Nalidixic acid, Ciprofloxacin, Pefloxacin and Ofloxacin determined by agar plate diffusion technique stood at 52.6, 47.3, 50.5, 70.6 and 46.0% for Escherichia coli, Salmonella sp., Shigellasp., Klebsiella sp. and Aeromonas sp. respectively. Analysis of variance indicated the occurrence of significant differences (F, 46.77-613.30; 0.00) in the resistance to each tested Qs. Generally, Human isolates showed greater resistance than Animal isolates (57.4 vs 47.2%). Investigation with specific primers indicated 11, 15, 7, 1 and 0 for qnrA, qnrB, qnrS, qepA and Aac(6')-Ib-cr genes respectively, out of 1018 Q-r and 29 PMQR isolates. Gene transfer experiments indicated the transfer of all genes except qepA either by conjugation or transformation. The MIC of tested Qs on recipient bacterium before gene transfer greatly increased from 0.0625 to 0.25 µg/mL, after transfer. This study demonstrates that PMQR genes amongst enteric bacteria in the Niger delta of Nigeria were transferable and transfer conferred a higher Q- resistance on recipient bacterium.202134250375
2046120.9998QRDR mutations, efflux system & antimicrobial resistance genes in enterotoxigenic Escherichia coli isolated from an outbreak of diarrhoea in Ahmedabad, India. BACKGROUND & OBJECTIVES: Diverse mechanisms have been identified in enteric bacteria for their adaptation and survival against multiple classes of antimicrobial agents. Resistance of bacteria to the most effective fluoroquinolones have increasingly been reported in many countries. We have identified that most of the enterotoxigenic Escherichia coli (ETEC) were resistant to several antimicrobials in a diarrhoea outbreak at Ahmedabad during 2000. The present study was done to identify several genes responsible for antimicrobial resistance and mobile genetic elements in the ETEC strains. METHODS: Seventeen ETEC strains isolated from diarrhoeal patients were included in this study. The antimicrobial resistance was confirmed by conventional disc diffusion method. PCR and DNA sequencing were performed for the identification of mutation in the quinolone resistance-determining regions (QRDRs). Efflux pump was tested by inhibiting the proton-motive force. DNA hybridization assay was made for the detection of integrase genes and the resistance gene cassettes were identified by direct sequencing of the PCR amplicons. RESULTS: Majority of the ETEC had GyrA mutations at codons 83 and 87 and in ParC at codon 80. Six strains had an additional mutation in ParC at codon 108 and two had at position 84. Plasmid-borne qnr gene alleles that encode quinolone resistance were not detected but the newly described aac(6')-Ib-cr gene encoding a fluoroquinolne-modifying enzyme was detected in 64.7 per cent of the ETEC. Class 1 (intI1) and class 2 (intI2) integrons were detected in six (35.3%) and three (17.6%) strains, respectively. Four strains (23.5%) had both the classes of integrons. Sequence analysis revealed presence of dfrA17, aadA1, aadA5 in class 1, and dfrA1, sat1, aadA1 in class 2 integrons. In addition, the other resistance genes such as tet gene alleles (94.1%), catAI (70.6%), strA (58.8%), bla TEM-1 (35.2%), and aphA1-Ia (29.4%) were detected in most of the strains. INTERPRETATION & CONCLUSIONS: Innate gene mutations and acquisition of multidrug resistance genes through mobile genetic elements might have contributed to the emergence of multidrug resistance (MDR) in ETEC. This study reinforces the necessity of utilizing molecular techniques in the epidemiological studies to understand the nature of resistance responsible for antimicrobial resistance in different species of pathogenic bacteria.201121911975
2906130.9998The mef(A) gene predominates among seven macrolide resistance genes identified in gram-negative strains representing 13 genera, isolated from healthy Portuguese children. Of the 176 randomly selected, commensal, gram-negative bacteria isolated from healthy children with low exposure to antibiotics, 138 (78%) carried one or more of the seven macrolide resistance genes tested in this study. These isolates included 79 (91%) isolates from the oral cavity and 59 (66%) isolates from urine samples. The mef(A) gene, coding for an efflux protein, was found in 73 isolates (41%) and was the most frequently carried gene. The mef(A) gene could be transferred from the donors into a gram-positive E. faecalis recipient and a gram-negative Escherichia coli recipient. The erm(B) gene transferred and was maintained in the E. coli transconjugants but was found in 0 to 100% of the E. faecalis transconjugants tested, while the other five genes could be transferred only into the E. coli recipient. The individual macrolide resistance genes were identified in 3 to 12 new genera. Eight (10%) of the oral isolates and 30 (34%) of the urine isolates for which the MICs were 2 to >500 microg of erythromycin per ml did not hybridize with any of the seven genes and may carry novel macrolide resistance genes.200415328110
2922140.9998Tetracycline-resistance genes in gram-negative isolates from estuarine waters. AIMS: To investigate the diversity and dissemination of tetracycline resistance genes in isolates from estuarine waters. METHODS AND RESULTS: Forty-two out of 164 multi-resistant isolates previously obtained were resistant or less-susceptible to tetracycline, as evaluated by the disc diffusion method. Minimal inhibitory concentration for resistant bacteria ranged from 16 to 256 mg l(-1). Screening of tet genes by polymerase chain reaction showed that 88% of the isolates carried at least one of the genes tested, namely tet(A) (present in 13 isolates), tet(B) (present in 13 isolates), tet(C) (present in 3 isolates), tet(D) (present in 1 isolate), tet(E) (present in 6 isolates) and tet(M) (present in 1 isolate). One isolate carried tet(A) and tet(M). To our knowledge, this study presents the first description of a tet(D) gene in Morganella morganii. Hybridization revealed that tet genes were plasmid-located in 31% of the isolates. Those isolates were included as donors in conjugation experiments and 38% transferred tetracycline resistance. CONCLUSIONS: A considerable diversity of tet genes was detected in the estuary. Frequently, these genes were associated with plasmids and could be transferred to Escherichia coli. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented provide further evidence of the role played by estuarine reservoirs in antibiotic resistance maintenance and dissemination.200819120920
895150.9998The determination of gyrA and parC mutations and the prevalence of plasmid-mediated quinolone resistance genes in carbapenem resistant Klebsiella pneumonia ST11 and ST76 strains isolated from patients in Heilongjiang Province, China. BACKGROUND: There is increasing resistance to carbapenems among Klebsiella pneumoniae,and fluoroquinolones (FQ) are increasingly used to treat infections from extended-spectrum β- lactamase(ESBLs) and carbapenemase-producing Klebsiella pneumoniae. However, the acquisition of plasmid-mediated quinolone resistance (PMQR) or the spontaneous mutation of the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes can severely affect the therapeutic effect of quinolones. The goal of this study was to investigate the molecular determinants of FQ resistance(FQ-R) in carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates from Heilongjiang Province,China. MATERIALS AND METHODS: We isolated 40 strains of CRKP from a treatment center in the eastern part of Heilongjiang Province from January 2016 to December 2018. The VITEK2 Compact analyzer was used to identify and detect drug sensitivity. Different types of drug resistance genes were detected by polymerase chain reaction (PCR). PCR and DNA sequencing were used to assess the presence of qnrA, qnrB, qnrS,qepA and acc(6') Ib-cr genes,which are plasmid-encode genes that can contribute to resistance. The sequences of gyrA and parC genes were sequenced and compared with the sequences of standard strains to determine if mutations were present.Multi-site sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were performed on the strains to assess homology. RESULTS: The isolated CRKP strains showed rates of resistance to fluoroquinolones of 22.5% to 42.5%. The resistance rate of ciprofloxacin was significantly higher than that of levofloxacin.Nine CRKP strains (22.5%) showed co-resistance to ciprofloxacin and levofloxacin.The quinolone resistant strains were screened for plasmid-encoded genes that can contribute to resistance (PMQR genes).Among the 17 quinolone resistant strains,one strain contained no PMQR genes,twelve strains contained two PMQR genes,and four strains contained four PMQR genes.Acc (6') Ib-cr was the most frequently detected PMQR gene, detected in 95% of strains tested (38 of 40) and in 94.1% of the quinolone-resistant strains (16 of 17). The qepA gene encoding an efflux pump was not detected in any strains.No isolate carried five different PMQRs simultaneously.Changes of S83I and D87G changes in gyrA, and the S80I change in parC,which were mediated by QRDR,were identified in two isolates,which showed resistance to both ciprofloxacin and levofloxacin.Most of the FQ-R strains(58.8%,10/17) belong to ST(sequence type) 76, which is dominant in the local area, while all the mutant strains (100%,2/2),that differ in at least one site from standard bacteria, belong to the ST11 group. The strains were isolated from a hospital where there had been a recent outbreak of ST76 type CRKP in the neurosurgery ward and intensive care unit. CONCLUSION: CRKP strains were identified that were insensitive or even resistant to quinolones,and this resistance is common in Heilongjiang Province of eastern China;fluoroquinolone-resistance in these clinical CRKP strains is a complex interplay between PMQR determinants and mutations in gyrA and parC.The resistance level caused by QRDR mutation is higher than that caused by PMQR, however, the high frequency of PMQR genes in the isolated CRKP strains suggests the potential for impact of these genes.PMQR determinants are often found in carbapenemase-producing or ESBLs-producing Klebsiella pneumoniae,and some resistance genes,such as:SHV,TEM, CTX-M-15,and OXA-1 are closely associated with FQ-R. Finally, geographical factors can affect the emergence and spread of PMQR and QRDR.Some genetic lineages have higher potential risks, and continuous close monitoring is required.202032278145
2023160.9998Class 1 and class 2 integrons and plasmid-mediated antibiotic resistance in coliforms isolated from ten rivers in northern Turkey. We aimed to determine the molecular mechanisms of antibiotic resistance in coliforms isolated from ten rivers in northern region of Turkey. A total of 183 isolates were tested for antimicrobial susceptibility by disk diffusion and agar dilution methods. Resistance to ampicillin, streptomycin, trimethoprim, tetracycline, and chloramphenicol was detected in 58%, 51.9%, 24%, 28.4%, and 12.5%, respectively. Twelve (6.5%) phylogenetically distant organisms were detected to harbor self-transmissible plasmids ranging 52 to >147 kb in sizes. Resistances to ampicillin, tetracycline, trimethoprim, streptomycin, and nalidixic acid were commonly transferable traits. Transferable nalidixic acid-resistant strains harbored qnrS gene, which was the first report of plasmid-mediated quinolone resistance in bacteria of environmental origin in Turkey. Fourteen and five coliforms harbored class 1 and class 2 integrons, respectively, and some of them were located on transferable plasmids. Sequence analyses of variable regions of the class 1 and 2 integrons harbored various gene cassettes, dfrA1, dfr2d, dfrA7, dfrA16, dfrA17, aadA1, aadA5, bla(oxA-30), and sat1. A gene cassette array, dfrA16 has been demonstrated for the first time in a Citrobacter koseri isolate. Class 1 and class 2-bearing strains were clustered in different groups by BOX-PCR fingerprinting. Rivers in the northern Turkey may act as receptacle for the multi-drug resistant enterobacteria and can serve as reservoirs of the antimicrobial resistance determinants in the environment. The actual risk to public health is the transfer of resistance genes from the environmental bacteria to human pathogens.200919229487
969170.9998Dissemination of the rmtB gene carried on IncF and IncN plasmids among Enterobacteriaceae in a pig farm and its environment. OBJECTIVES: To investigate the prevalence and characterization of 16S rRNA methylase-producing bacteria in a pig farm and its environment in East China. METHODS: Enterobacteriaceae isolates and metagenomic DNA from 102 pig faecal samples from a pig farm and 97 soil samples taken in or around the farm were screened for the presence of 16S rRNA methylase genes. The clonal relationships of 16S rRNA methylase-positive isolates, plasmid content and other associated resistance genes were also characterized. RESULTS: Fifty-six rmtB-positive Enterobacteriaceae isolates, including 54 Escherichia coli, 1 Morganella morganii and 1 Proteus mirabilis, were recovered from 55 pig faecal samples. Nineteen rmtB-positive bacteria, including 13 E. coli, 2 M. morganii, 2 Leclercia adecarboxylata, 1 Enterobacter aerogenes and 1 Enterobacter cloacae, were recovered from 16 soil samples. Among the 75 rmtB-positive isolates, 31 and 25 also carried the qepA and bla(CTX-M) genes, respectively. The qepA gene co-localized with rmtB on the F2:A-:B1 plasmids and the bla(CTX-M-65) gene co-localized with rmtB on the F33:A-:B- plasmids. The rmtB gene was also found to be associated with the IncN plasmids. Clonal transmission of rmtB-positive E. coli isolates was observed between different pig groups and soil samples. CONCLUSIONS: Both horizontal gene transfer and clonal spread could be responsible for the dissemination of the rmtB gene in the pig farm and its environment. To our knowledge, this study is the first report of rmtB-positive bacteria from farmland soils and indicates that these antibiotic-resistant bacteria and/or resistance genes could be acquired by humans through the food chain.201121852287
967180.9998Characterization of Integrons and Quinolone Resistance in Clinical Escherichia coli Isolates in Mansoura City, Egypt. Escherichia coli is a common pathogen in both humans and animals. Quinolones are used to treat infections caused by Gram-negative bacteria, but resistance genes emerged. Only scarce studies investigated the association between plasmid-mediated quinolone resistance (PMQR) genes and integrons in clinical isolates of E. coli. The current study investigated the prevalence of quinolone resistance and integrons among 134 clinical E. coli isolates. Eighty (59.70%) isolates were quinolone-resistant, and 60/134 (44.77%) isolates were integron positive with the predominance of class I integrons (98.33%). There was a significant association between quinolone resistance and the presence of integrons (P < 0.0001). Isolates from Urology and Nephrology Center and Gastroenterology Hospital were significantly quinolone-resistant and integron positive (P ≤ 0.0005). Detection of PMQR genes on plasmids of integron-positive isolates showed that the active efflux pump genes oqxAB and qepA had the highest prevalence (72.22%), followed by the aminoglycoside acetyltransferase gene (aac(6')-Ib-cr, 66.67%) and the quinolone resistance genes (qnr, 61.11%). Amplification and sequencing of integrons' variable regions illustrated that no quinolone resistance genes were detected, and the most predominant gene cassettes were for trimethoprim and aminoglycoside resistance including dfrA17, dfrB4, and dfrA17-aadA5. In conclusion, this study reported the high prevalence of PMQR genes and integrons among clinical E. coli isolates. Although PMQR genes are not cassette-born, they were associated with integrons' presence, which contributes to the widespread of quinolone resistance in Egypt.202134527054
2981190.9998Investigation of plasmid-mediated resistance in E. coli isolated from healthy and diarrheic sheep and goats. Escherichia coli is zoonotic bacteria and the emergence of antimicrobial-resistant strains becomes a critical issue in both human and animal health globally. This study was therefore aimed to investigate the plasmid-mediated resistance in E. coli strains isolated from healthy and diarrheic sheep and goats. A total of 234 fecal samples were obtained from 157 sheep (99 healthy and 58 diarrheic) and 77 goats (32 healthy and 45 diarrheic) for the isolation and identification of E. coli. Plasmid DNA was extracted using the alkaline lysis method. Phenotypic antibiotic susceptibility profiles were determined against the three classes of antimicrobials, which resistance is mediated by plasmids (Cephalosporins, Fluoroquinolone, and Aminoglycosides) using the disc-diffusion method. The frequency of plasmid-mediated resistance genes was investigated by PCR. A total of 159 E. coli strains harbored plasmids. The isolates antibiogram showed different patterns of resistance in both healthy and diarrheic animals. A total of (82; 51.5%) E. coli strains were multidrug-resistant. rmtB gene was detected in all Aminoglycoside-resistant E. coli, and the ESBL-producing E. coli possessed different CTX-M genes. Similarly, fluoroquinolone-resistant E. coli possessed different qnr genes. On the analysis of the gyrB gene sequence of fluoroquinolone-resistant E. coli, multiple point mutations were revealed. In conclusion, a high prevalence of E. coli with high resistance patterns to antimicrobials was revealed in the current study, in addition to a wide distribution of their resistance determinants. These findings highlight the importance of sheep and goats as reservoirs for the dissemination of MDR E. coli and resistance gene horizontal transfer.202032127753