Beta-lactams resistance and presence of class 1 integron in Pseudomonas spp. isolated from untreated hospital effluents in Brazil. - Related Documents




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103101.0000Beta-lactams resistance and presence of class 1 integron in Pseudomonas spp. isolated from untreated hospital effluents in Brazil. The aim of the present study was to investigate the resistance profile, to detect the presence of beta-lactam resistance genes, phenotypic expression of efflux pump systems and class 1 integrons in Pseudomonas spp. strains obtained from untreated hospital effluents. Effluent samples were collected from four hospitals in Porto Alegre, RS, Brazil. Pseudomonas were isolated on MacConkey agar plates and the identification was confirmed by 16S rRNA PCR and biochemical tests. Susceptibility testing was determined by disk-diffusion method using 11 different beta-lactams and MIC assays were performed on isolates resistant to imipenem and ceftazidime. The beta-lactamase genes bla (IMP), bla (VIM), bla (SPM-1), bla (OXA-23-like), bla (OXA-24-like), bla (OXA-51-like) and the intl1 gene from class 1 integron were analysed by PCR. One hundred and twenty-four isolates were recovered and the most common species was Pseudomonas pseudoalcaligenes. The resistance found among the isolates was considered high, 62 (50%) isolates were multiresistant. No isolate carrying the beta-lactamase genes tested was found among the strains. Seven isolates showed reduction of MIC for imipenem and ceftazidime in the presence of cyanide m-chlorophenylhydrazone, indicating the hyper expression of efflux pumps. From the 124 isolates, 52 (41.9%) were identified as carrying the class 1 integron gene, intI1. Untreated hospital effluents could be a source of environmental contamination due to discharge of antimicrobial resistant bacteria which can carry integron class 1 and act as a reservoir of resistance genes and have efflux pump systems.201222382676
103410.9999Detection of metallo-beta-lactamase-producing genes bla(SPM) and bla(NDM) in Pseudomonas aeruginosa isolated from wastewater in Southern Brazil. Pseudomonas aeruginosa is commonly associated with the ability to acquire antimicrobial resistance. The surveillance of resistance genes in various environmental matrices has gained prominence in recent years, being seen as a potential threat to public health. The objective of this study was to investigate genes encoding metallo-beta-lactamases (MBLs), which confer resistance to carbapenems, in wastewater. Fifteen isolates of P. aeruginosa were collected for five months from samples obtained from a municipal wastewater treatment plant in Rio Grande do Sul. These isolates were subjected to disk diffusion testing using 10 different antimicrobials. Phenotypic enzymatic tests for MBLs were conducted, and positive isolates underwent DNA extraction and gene detection using the polymerase chain reaction. The resistance rate to ceftazidime was 100%, cefepime 73.3%, piperacillin-tazobactam 66.67%, imipenem 53.30%, levofloxacin 46.67%, tobramycin 40%, and ciprofloxacin and amikacin 13.33%. Both meropenem and aztreonam resistances were rare accounting for 6.60% of the tested isolates. Among these isolates, 20% were classified as multidrug-resistant and were found to carry the bla(NDM) and bla(SPM) genes. The results suggest that evaluating resistance genes in bacteria from urban raw sewage can provide data that assist in surveillance, as this environment can stimulate increased bacterial resistance.202438678422
103320.9999Antimicrobial Resistance and β-Lactamase Production in Clinically Significant Gram-Negative Bacteria Isolated from Hospital and Municipal Wastewater. Hospital and municipal wastewater contribute to the spread of antibiotic-resistant bacteria and genes in the environment. This study aimed to examine the antibiotic resistance and β-lactamase production in clinically significant Gram-negative bacteria isolated from hospital and municipal wastewater. The susceptibility of bacteria to antibiotics was tested using the disk diffusion method, and the presence of extended-spectrum β-lactamases (ESBL) and carbapenemases was determined using an enzyme inhibitor and standard multiplex PCR. Analysis of antimicrobial resistance of total bacterial strains (n = 23) revealed that most of them were resistant to cefotaxime (69.56%), imipenem (43.47%), meropenem (47.82%) and amoxicillin-clavulanate (43.47%), gentamicin (39.13%), cefepime and ciprofloxacin (34.78%), trimethoprim-sulfamethoxazole (30.43%). A total of 8 of 11 phenotypically confirmed isolates were found to have ESBL genes. The bla(TEM) gene was present in 2 of the isolates, while the bla(SHV) gene was found in 2 of the isolates. Furthermore, the bla(CTX-M) gene was found in 3 of the isolates. In one isolate, both the bla(TEM) and bla(SHV) genes were identified. Furthermore, of the 9 isolates that have been phenotypically confirmed to have carbapenemase, 3 were confirmed by PCR. Specifically, 2 isolates have the bla(OXA-48) type gene and 1 have the bla(NDM-1) gene. In conclusion, our investigation shows that there is a significant rate of bacteria that produce ESBL and carbapenemase, which can promote the spread of bacterial resistance. Identifying ESBL and carbapenemase production genes in wastewater samples and their resistance patterns can provide valuable data and guide the development of pathogen management strategies that could potentially help reduce the occurrence of multidrug resistance.202337107015
103230.9999Molecular investigation of antibiotic resistant bacterial strains isolated from wastewater streams in Pakistan. Antibiotic resistance is a global public health issue and it is even more daunting in developing countries. The main objective of present study was to investigate molecular responses of antibiotic-resistant bacteria. The 48 bacterial strains, which were previously isolated and identified were subjected to disc diffusion and MIC (minimum inhibitory concentration) determination, followed by investigating the production of the three beta-lactamases (ESBLs (Extended-spectrum Beta-lactamases), MBLs (Metallo Beta-lactamases), AmpCs) and exploring prevalence of the two antibiotic-resistant genes (ARGs); blaTEM and qnrS. Higher MIC values were observed for penicillin(s) than that for fluoroquinolones (ampicillin > amoxicillin > ofloxacin > ciprofloxacin > levofloxacin). Resistance rates were high (58-89%) for all of the tested beta-lactams. Among the tested strains, 5 were ESBL producers (4 Aeromonas spp. and 1 Escherichia sp.), 2 were MBL producers (1 Stenotrophomonas sp. and 1 Citrobacter sp.) and 3 were AmpC producers (2 Pseudomonas spp. and 1 Morganella sp.). The ARGs qnrS2 and blaTEM were detected in Aeromonas spp. and Escherichia sp. The results highlighted the role of Aeromonas as a vector. The study reports bacteria of multidrug resistance nature in the wastewater environment of Pakistan, which harbor ARGs of clinical relevance and could present a public health concern.202032802720
214240.9999Resistance to β-lactams and distribution of β-lactam resistance genes in subgingival microbiota from Spanish patients with periodontitis. OBJECTIVES: The aim of this study was to analyze the distribution of β-lactamase genes and the multidrug resistance profiles in β-lactam-resistant subgingival bacteria from patients with periodontitis. MATERIALS AND METHODS: Subgingival samples were obtained from 130 Spanish patients with generalized periodontitis stage III or IV. Samples were grown on agar plates with amoxicillin or cefotaxime and incubated in anaerobic and microaerophilic conditions. Isolates were identified to the species level by the sequencing of their 16S rRNA gene. A screening for the following β-lactamase genes was performed by the polymerase chain reaction (PCR) technique: bla(TEM), bla(SHV), bla(CTX-M), bla(CfxA), bla(CepA), bla(CblA), and bla(ampC). Additionally, multidrug resistance to tetracycline, chloramphenicol, streptomycin, erythromycin, and kanamycin was assessed, growing the isolates on agar plates with breakpoint concentrations of each antimicrobial. RESULTS: β-lactam-resistant isolates were found in 83% of the patients. Seven hundred and thirty-seven isolates from 35 different genera were obtained, with Prevotella and Streptococcus being the most identified genera. bla(CfxA) was the gene most detected, being observed in 24.8% of the isolates, followed by bla(TEM) (12.9%). Most of the isolates (81.3%) were multidrug-resistant. CONCLUSIONS: This study shows that β-lactam resistance is widespread among Spanish patients with periodontitis. Furthermore, it suggests that the subgingival commensal microbiota might be a reservoir of multidrug resistance and β-lactamase genes. CLINICAL RELEVANCE: Most of the samples yielded β-lactam-resistant isolates, and 4 different groups of bla genes were detected among the isolates. Most of the isolates were also multidrug-resistant. The results show that, although β-lactams may still be effective, their future might be hindered by the presence of β-lactam-resistant bacteria and the presence of transferable bla genes.202032495224
103650.9999Detection of carbapenem resistance genes and cephalosporin, and quinolone resistance genes along with oqxAB gene in Escherichia coli in hospital wastewater: a matter of concern. AIMS: This study was performed to detect the presence of Escherichia coli resistant to cephalosporins, carbapenems and quinolones in hospital wastewater. METHODS AND RESULTS: Wastewaters from a rural (H1) and an urban (H2) hospital were tested for E. coli resistant to cephalosporins, carbapenem and quinolones. Genes coding for chromosomal and plasmid-mediated resistance and phylogenetic grouping was detected by multiplex polymerase chain reaction (PCR) and for genetic relatedness by rep-PCR. Of 190 (H1 = 94; H2 = 96) E. coli examined, 44% were resistant to both cephalosporins and quinolones and 3% to imipenem. ESBLs were detected phenotypically in 96% of the isolates, the gene blaCTX-M coding for 87% and blaTEM for 63%. Quinolone resistance was due to mutations in gyrA and parC genes in 97% and plasmid-coded aac-(6')-Ib-cr in 89% of isolates. Only in one carbapenem-resistant E. coli, NDM-1 was detected. Nearly 67% of the isolates belonged to phylogenetic group B2. There was no genetic relatedness among the isolates. CONCLUSIONS: Hospital wastewater contains genetically diverse multidrug-resistant E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: This study stresses the need for efficient water treatment plants in healthcare settings as a public health measure to minimize spread of multidrug-resistant bacteria into the environment.201424975198
105760.9999Emergence of ciprofloxacin-resistant extended-spectrum β-lactamase-producing enteric bacteria in hospital wastewater and clinical sources. This study aimed to evaluate the incidence of ciprofloxacin-resistant extended-spectrum β-lactamase (ESBL)-producing enteric bacteria in hospital wastewater and clinical sources. Enteric bacteria, mainly Escherichia coli, were isolated from clinical sources (urinary tract and gastrointestinal tract infections; 80 isolates) and hospital wastewater (103 isolates). The antibiotic resistance profile and ESBL production of the isolates were investigated by disc diffusion assay and combined disc diffusion test, respectively. Plasmid profiling was performed by agarose gel electrophoresis, and elimination of resistance markers was performed by a plasmid curing experiment. Antibiotic susceptibility testing revealed a high incidence of β-lactam resistance, being highest to ampicillin (88.0%) followed by amoxicillin, ceftriaxone, cefpodoxime, cefotaxime, aztreonam, cefepime and ceftazidime. Among the non-β-lactam antibiotics, the highest resistance was recorded to nalidixic acid (85.7%). Moreover, 50.8% of enteric bacteria showed resistance to ciprofloxacin. Among 183 total enteric bacteria, 150 (82.0%) exhibited multidrug resistance. ESBL production was detected in 78 isolates (42.6%). A significantly higher incidence of ciprofloxacin resistance was observed among ESBL-producing enteric bacteria both in clinical (P=0.0015) and environmental isolates (P=0.012), clearly demonstrating a close association between ESBL production and ciprofloxacin resistance. Plasmid profiling of selected ESBL-positive strains indicated the presence of one or more plasmids of varying sizes. Plasmid curing resulted in loss of ciprofloxacin and cefotaxime resistance markers simultaneously from selected ESBL-positive isolates, indicating the close relationship of these markers. This study revealed a common occurrence of ciprofloxacin-resistant ESBL-producing enteric bacteria both in hospital wastewater and clinical sources, indicating a potential public health threat.201627436461
90070.9999Presence of OXA-23-producing isolates of Acinetobacter baumannii in wastewater from hospitals in southern Brazil. The aim of the study was to evaluate the dissemination of multiresistant isolates of Acinetobacter baumannii carrying resistance genes, by samples of wastewater from hospitals in Porto Alegre, Rio Grande do Sul, Brazil. We obtained 303 bacterial isolates from the wastewater of three hospitals in Porto Alegre, Rio Grande do Sul. For each isolate, we determined the profile of susceptibility to antimicrobials and the presence of the genes bla(OXA-23), bla(OXA-24), bla(OXA-51), bla(OXA-58), bla(SPM-1), bla(IMP), and bla(VIM.) The bla(OXA-51) gene was found in 56% of the isolates, indicating the presence of A. baumannii in this environment. Of these, three multiresistant isolates were positive for the bla(OXA-23) gene, in wastewater from two of the hospitals. The results obtained in this study indicate that isolates of A. baumannii which are multiresistant and carry resistance genes such as bla(OXA-51) and bla(OXA-23) are being released into the environment in the wastewater from the hospitals analyzed. Multiresistant Acinetobacter junii, the newly emerging pathogen, were also found among the multiresistant isolates. Hospital wastewater may be crucial to the development and dispersal of multiresistant bacteria, making waterbodies reservoirs of bacterial resistance.201121254809
102880.9998Antibiotic resistance and extended-spectrum β-lactamases in isolated bacteria from seawater of Algiers beaches (Algeria). The aim of the study was to evaluate bacterial antibiotic resistance in seawater from four beaches in Algiers. The most significant resistance rates were observed for amoxicillin and ticarcillin, whereas they were relatively low for ceftazidime, cefotaxime and imipenem. According to sampling sites, the highest resistance rates were recorded for 2 sites subjected to chemical and microbiological inputs (amoxicillin, 43% and 52%; ticarcillin, 19.6% and 47.7%), and for 2 sites relatively preserved from anthropogenic influence, resistance rates were lowest (amoxicillin, 1.5% and 16%; ticarcillin, 0.8% and 2.6%). Thirty-four bacteria resistant to imipenem (n=14) or cefotaxime (n=20) were identified as Pseudomonas aeruginosa (n=15), Pseudomonas fluorescens (7), Stenotrophomonas maltophilia (4), Burkholderia cepacia (2), Bordetella sp. (1), Pantoea sp. (1), Acinetobacter baumannii (1), Chryseomonas luteola (1), Ochrobactrum anthropi (1) and Escherichia coli (1). Screening for extended spectrum β-lactamase showed the presence of CTX-M-15 β-lactamase in the E. coli isolate, and the encoding gene was transferable in association with the IncI1 plasmid of about 50 kbp. Insertion sequence ISEcp1B was located upstream of the CTX-M-15 gene. This work showed a significant level of resistance to antibiotics, mainly among environmental saprophytic bacteria. Transmissible CTX-M-15 was detected in E. coli; this may mean that contamination of the environment by resistant bacteria may cause the spread of resistance genes.201222095134
117790.9998High carriage of plasmid-mediated quinolone resistance (PMQR) genes by cefotaxime-resistant Escherichia coli recovered from surface-leaking sanitary sewers. There is a rapid rise in the incidence of quinolone resistant bacteria in Nigeria. Most studies in Nigeria have focused on isolates from the clinical settings, with few focusing on isolates of environmental origin. This study aimed to investigate the antibiogram and carriage of plasmid-mediated quinolone resistance (PMQR) genes by quinolone-resistant isolates obtained from a pool of cefotaxime-resistant Escherichia coli (E. coli) recovered from sewage leaking out of some surface-leaking sanitary sewers in a University community in Nigeria. Isolation of E. coli from the sewage samples was done on CHROMagar E. coli, after enrichment of the samples was done in Brain Heart Infusion broth amended with 6 µg/mL of cefotaxime. Identification of presumptive E. coli was done using molecular methods (detection of uidA gene), while susceptibility to antibiotics was carried out using the disc diffusion method. Detection of PMQR genes (qnrA, qnrB, qnrS, aac(6')-lb-cr, qepA and oqxAB) was carried out using primer-specific PCR. A total of 32 non-repetitive cefotaxime-resistant E. coli were obtained from the sewage, with 21 being quinolone-resistant. The quinolone-resistant isolates showed varying level of resistance to the tested antibiotics, with imipenem being the only exception with 0% resistance. The PMQR genes: aac(6')-lb-cr, qnrA, qnrB, qnrS and qepA and oqxAB were detected in 90.5%, 61.9%, 47.6%, 38.1%, 4.8% and 0% respectively of the isolates. The findings of this study showed a high level of resistance to antibiotics and carriage of PMQR genes by quinolone-resistant E. coli obtained from the leaking sanitary sewers, suggesting a potential environmental and public health concern.202235000007
1015100.9998Antimicrobial-resistant and extended-spectrum β-lactamase-producing Escherichia coli in raw cow's milk. The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)-producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.201525581180
897110.9998Prevalence of class 1 integrons and plasmid-mediated qnr-genes among Enterobacter isolates obtained from hospitalized patients in Ahvaz, Iran. Quinolones are frequently used classes of antimicrobials in hospitals, crucial for the treatment of infections caused by Gram-negative bacteria. The inappropriate use of quinolones and other antimicrobial agents for the treatment of bacterial infections leads to a significant increase of resistant isolates. The acquisition of antimicrobial resistance may be related to achievement of resistance determinant genes mediated by plasmids, transposons and gene cassettes in integrons. The objective of this cross-sectional study, conducted from December 2015 to July 2016 at two teaching hospitals in Ahvaz, southern Iran, was to screen for the presence of class 1 integrons and quinolone resistance genes in clinical isolates of Enterobacter spp. In all, 152 non-duplicated Enterobacter isolates were collected from clinical specimens and identified as Enterobacter spp. using standard microbiological methods. Antimicrobial susceptibility test was determined using the disc diffusion method according to the CLSI recommendation. Determination of class 1 integrons and PMQR genes was assessed by PCR. Analysis of antibiotic susceptibility tests showed that the highest antibiotic resistance was toward ciprofloxacin (55.3%), while the lowest level was observed against meropenem (34.9%). Moreover, 47.4% (72/152) and 29% (44/152) of isolates were positive for class 1 integron and quinolone resistance genes, respectively. The relative frequencies of antibiotic resistance were significantly higher among class 1 integron-positive isolates. In summary, our results highlight the importance of PMQR genes in the emergence of quinolone-resistant Enterobacter isolates. Moreover, it seems that class 1 integrons have a widespread distribution among Enterobacter isolates and have clinical relevance to multiple-drug-resistant isolates.201729286015
968120.9998Molecular 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
1109130.9998Quinolone Susceptibility and Detection of qnr and aac(6')-Ib-cr Genes in Community Isolates of Klebsiella pneumoniae. BACKGROUND: Plasmid-mediated quinolone resistance genes (PMQR) have been shown to play not only an important role in quinolone resistance, but also resistance to other antibiotics, particularly β-lactams and aminoglycosides. These genes are mainly associated with clinical isolates of Enterobacteriaceae. However, detection of PMQR genes in the community isolates can increase the dissemination rate of resistance determinants among bacteria. OBJECTIVES: This study aimed to investigate quinolone resistance and distribution of qnr and aac (6')-Ib-cr genes among the community isolates of Klebsiella pneumoniae. MATERIALS AND METHODS: Fifty-two K. pneumoniae isolates were collected from the Central Laboratory in Karaj between July 2010 and January 2011. Antibacterial susceptibility was determined by the disc diffusion method. Quinolone and/or cephalosporin-resistant isolates were screened for the presence of qnrA, qnrB, qnrS and aac (6')-Ib-cr genes by polymerase chain reaction (PCR). RESULTS: Of the 52 K. pneumoniae isolates, 23 were resistant to cephalosporins and/or quinolones. Overall, 7 out of the 23 resistant isolates harbored qnr and/or aac (6')-Ib-cr genes (30.4%). Among these, 5 isolates were resistant to both classes of antibiotics of which; 3 carried the aac (6')-Ib-cr gene, one had the qnrS, and one harbored both aac (6')-Ib-cr and qnrB genes. None of the isolates contained qnrA. Two isolates were sensitive to quinolones and resistant to cephalosporins of which; one had qnrS and the other carried the aac (6')-Ib-cr gene. CONCLUSIONS: Our study showed that 30.4% of the quinolone and/or cephalosporin resistant community isolates of K. pneumoniae carried PMQR genes. These results confirm that community isolates can be an important source for spreading antibiotic resistance determinants among Gram negative pathogens. This is the first report from Iran on detection of PMQR in the community isolates of K. pneumoniae.201425368793
1058140.9998First Detection of FOX-1 AmpC β-lactamase Gene Expression Among Escherichia coli Isolated from Abattoir Samples in Abakaliki, Nigeria. OBJECTIVES: Gram-negative bacteria represent the most relevant reservoir of resistance to antibiotics in the environment. The natural selection of resistant clones of bacteria in the environment by antimicrobial selective pressure is a relevant mechanism for spreading antibiotic resistance traits in both the community and hospital environment. This is in scenarios where antimicrobials are used irrationally, and even in the propagation of livestock, poultry birds, and for other veterinary purposes. This study sought to detect the prevalence of FOX-1 AmpC β-lactamase genes from abattoir samples. METHODS: The isolation of Escherichia coli, antimicrobial susceptibility testing, and β-lactamase characterization was carried out using standard microbiology techniques. The production of AmpC β-lactamase was phenotypically carried out using the cefoxitin-cloxacillin double-disk synergy test (CC-DDST), and FOX-1 AmpC genes was detected in the E. coli isolates using multiplex polymerase chain reaction. RESULTS: Forty-eight E. coli isolates were recovered from the anal swabs of cows and 35 (72.9%) isolates were positive for the production of β-lactamase. Notably, high percentages of resistance to cefoxitin (91.7%), ceftriaxone (83.3%), imipenem (85.4%), ceftazidime (87.5%), ofloxacin (81.3%), and gentamicin (85.4%) were found. FOX-1 genes were detected in three (6.3%) of the 48 E. coli isolates phenotypically screened for AmpC enzyme production. CONCLUSIONS: Abattoirs could represent a major reservoir of resistance genes especially AmpC β-lactamase, and this could serve as a route for the dissemination of multidrug-resistant bacteria in the community. Thus, the molecular identification of drug-resistant genes is vital for a reliable epidemiological investigation and the forestalling of the emergence and spread of these organisms through the food chain in this region.201829896333
1029150.9998Phylogenetic relationships, virulence and antimicrobial resistance properties of Klebsiella sp. isolated from pet turtles in Korea. Klebsiella sp. are responsible for a multitude of infectious diseases in both humans and animals. In this study, phylogenetic relationships, virulence and antimicrobial resistance gene properties of 16 Klebsiella sp. isolated from 49 pet turtles were investigated. The isolates including Klebsiella oxytoca (n = 13) and Klebsiella pneumoniae (n = 3) were identified using 16S rRNA gene sequencing and each species formed distinct clusters in the neighbour-joining phylogenetic tree. The prevalence of virulence genes including ureC (100%) and kfu (68·75%) was observed among the isolates using Polymerase chain reaction (PCR) assay. The fimH, mrkD and rmpA genes were detected in all K. pneumoniae while these were absent in every K. oxytoca isolate. In antimicrobial susceptibility testing, high resistance rates were observed against ampicillin (100%) and cephalothin (62·50%). The resistance rates against imipenem, tetracycline, trimethoprim/sulfamethoxazole, nalidixic acid and ciprofloxacin were 12·50, 12·50, 12·50, 6·25 and 6·25% respectively. The presence of antimicrobial resistance genes such as plasmid-mediated quinolone resistance (PMQR) [qnrB (37·50%), qnrA (31·25%), qnrS (12·50%) and aac(6')-Ib-cr (12·50%)], extended-spectrum β-lactamase (ESBL) [bla(CTX-M) (18·75%)], β-lactamase [bla(SHV-1) (18·75%)] and tetracycline resistance [tetE (12·50%)] was observed. The results revealed that pet turtle-borne Klebsiella sp. may carry different types of virulence and antimicrobial resistance genes which represents a potential threat to public health. SIGNIFICANCE AND IMPACT OF THE STUDY: Klebsiella sp. are nonmotile Gram-negative bacteria that are found in different environments. The virulence and antimicrobial resistance properties of pet turtle-borne Klebsiella sp. have not been studied before. Phylogenetic relationships, virulence traits and antimicrobial resistance profiles of pet turtle-borne Klebsiella sp. were characterized for the first time in Korea. Multiple virulence and antimicrobial resistance genes were observed among the isolates. The occurrence of virulence and antimicrobial resistance determinants in Klebsiella sp. may represent a potential threat to public health.202031671218
923160.9998Prevalence of Oxacillinase Genes in Clinical Multidrug-Resistant Gram-Negative Bacteria. BACKGROUND: The emergence of OXA-type beta-lactamases has become a significant threat to public healthcare systems and may lead to prolonged hospital stays and increased mortality rates among affected patients. This study aimed to determine the prevalence of oxacillinase resistance (OXA) genes in multidrug-resistant (MDR) Gram-negative bacteria. METHODS: One hundred and six clinical isolates were collected from a stock of Gram-negative isolates and were identified and tested for antibiotic susceptibility and presence of OXA genes using polymerase chain reaction (PCR). RESULTS: The most common detected isolate was Klebsiella pneumoniae (36.8%), followed by Escherichia coli (33%), Pseudomonas aeruginosa (16%), and Acinetobacter baumannii (14.2%). Out of these isolates, 97.4%, 87.2%, 84.6%, and 79.5% were resistant to ampicillin/sulbactam, cefotaxime, ceftazidime, and aztreonam, respectively. PCR results confirmed the presence of one or more OXA genes in 34% of the samples studied. The blaOXA-1 and blaOXA-10 genes were the most highly detected genes, followed by blaOXA-4 and blaOXA-51. The total number of Pseudomonas aeruginosa isolates was confirmed to carry at least one OXA gene (70.6%), whereas Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli were confirmed to carry at least one OXA gene (53.3, 28.2, and 22.9%, respectively). There was a significant association (p < 0.05) between the resistance genes and the type of isolate. CONCLUSIONS: Pseudomonas aeruginosa and Acinetobacter baumannii are the most common MDR Gram-negative strains carrying OXA-type beta-lactamase genes. Monitoring of MDR pathogens in Gram-negative bacteria must be continuously undertaken to implement effective measures for infection control and prevention.202540066541
1038170.9998Isolation of Extended Spectrum β-lactamase (ESBL) Producing Bacteria from Urban Surface Waters in Malaysia. BACKGROUND: This was a preliminary study to test for the presence of multiple antibiotic-resistant extended spectrum β-lactamase (ESBL) producing bacteria in Malaysian urban surface waters. Although the literature review revealed several published papers on clinical ESBL isolates in Malaysia, none were found on ESBL isolates obtained from local surface waters. METHODS: Isolated bacterial species were tested for resistance to cefotaxime, amoxicillin/clavulanate and aztreonam, and susceptibility to imipenem and meropenem using antibiotic susceptibility testing (AST) by disc diffusion. This served as a screening step to detect bacteria that could be potential ESBL species. 16S ribose ribonucleic acid (rRNA) polymerase chain reaction (PCR) testing with two clusters of bla (β-lactamase) gene primers was used to test for the bla genes CTX-M (Groups 1, 2, 9), OXA-1, SHV and TEM. RESULTS: A total of 19 isolates were found, possessing at least one of the bla genes tested for. There was a relatively high occurrence of CTX-M genes (84.2%) among these, followed by TEM genes (47.4%). The isolates were identified as Enterobacteriaceae (89.5%), predominantly Escherichia coli and Klebsiella pneumoniae. CONCLUSION: There appears to be a high occurrence of ESBL-bacteria in local surface waters, among these being opportunistic pathogens. The persistence and spread of these species in the environment poses a threat to exposed human populations.201323966820
1178180.9998Molecular Characterization of Plasmid-Mediated Quinolone Resistance Genes in Multidrug-Resistant Escherichia coli Isolated From Wastewater Generated From the Hospital Environment. AIM: This study investigated the carriage of Plasmid-Mediated Quinolone Resistance (PMQR) genes in fluoroquinolone-resistant Escherichia coli recovered from wastewater generated by healthcare institutions. MATERIALS AND METHODS: Isolation of fluoroquinolone-resistant Escherichia coli was done on medium supplemented with 1 µg/mL of ciprofloxacin (a fluoroquinolone). Presumptive isolates were identified via the detection of uidA gene. Susceptibility of the isolates to a panel of antibiotics was done using disc diffusion method. Detection of PMQR genes in the isolates was done using primer-specific PCR. RESULTS: Thirty fluoroquinolone-resistant Escherichia coli were obtained from the wastewater over a period of 6 months. The resistance to each of the antibiotic tested was: ampicillin (100%), ceftriaxone (100%), nalidixic acid (100%), tetracycline (96.7%), cefotaxime (96.7%), amoxicillin-clavulanate (80%), gentamicin (60%), cefoxitin (30%), and imipenem (3.3%). The Multiple Antibiotic Resistance Index (MARI) ranged from 0.6 to 0.9. The detection of PMQR genes in the 30 isolates was: qnrA (76.7%), qnrB (53.3%), qnrS (63.3%), aac(6')-lb-cr (43.3%), and qepA (43.3%). All the fluoroquinolone-resistant Escherichia coli carried at least one PMQR determinant. CONCLUSION: This study revealed that untreated hospital wastewaters are significant hub of multidrug-resistant and fluoroquinolone-resistant Escherichia coli, showing high carriage of PMQR genes, and may be a major contributor to the resistome of fluoroquinolone-resistant bacteria in the Nigerian environment.202540552214
1054190.9998Molecular detection of extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates of chicken origin from East Java, Indonesia. BACKGROUND AND AIM: Klebsiella pneumoniae is one of the respiratory disease agents in human and chicken. This bacterium is treated by antibiotic, but this treatment may trigger antibiotic resistance. Resistance gene in K. pneumoniae may be transferred to other bacteria. One of the known resistance genes is extended-spectrum β-lactamase (ESBL). This research aimed to study K. pneumoniae isolated from chicken farms in East Java, Indonesia, by observing the antibiotic resistance pattern and detect the presence of ESBL coding gene within the isolates. MATERIALS AND METHODS: A total of 11 K. pneumoniae isolates were collected from 141 chicken cloacal swabs from two regencies in East Java. All isolates were identified using the polymerase chain reaction method. Antimicrobial susceptibility was determined by agar dilution method on identified isolates, which then processed for molecular characterization to detect ESBL coding gene within the K. pneumoniae isolates found. RESULTS: The result of antibiotic sensitivity test in 11 isolates showed highest antibiotic resistance level toward ampicillin, amoxicillin, and oxytetracycline (100%, 100%, and 90.9%) and still sensitive to gentamicin. Resistance against colistin, doxycycline, ciprofloxacin, and enrofloxacin is varied by 90.9%, 54.5%, 27.3%, and 18.2%, respectively. All isolates of K. pneumoniae were classified as multidrug resistance (MDR) bacteria. Resistance gene analysis revealed the isolates harbored as bla (SHV) (9.1%), bla (TEM) (100%), and bla (CTX-M) (90.9%). CONCLUSION: All the bacterial isolates were classified as MDR bacteria and harbored two of the transmissible ESBL genes. The presence of antibiotic resistance genes in bacteria has the potential to spread its resistance properties.201931190714