Antimicrobial Susceptibility Profiles among Pseudomonas aeruginosa Isolated from Professional SCUBA Divers with Otitis Externa, Swimming Pools and the Ocean at a Diving Operation in South Africa. - Related Documents




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121701.0000Antimicrobial Susceptibility Profiles among Pseudomonas aeruginosa Isolated from Professional SCUBA Divers with Otitis Externa, Swimming Pools and the Ocean at a Diving Operation in South Africa. SCUBA divers are predisposed to otitis externa caused by Pseudomonas aeruginosa, which is becoming increasingly multi-drug resistant (MDR). The present work assessed the antibiotic resistance profiles of P. aeruginosa obtained from SCUBA divers and their environment in Sodwana Bay, South Africa. Bacterial isolates from a total of 137 random water and ear swab samples were identified using biochemical and molecular methods. P. aeruginosa strains were further evaluated for antibiotic susceptibility using the Kirby-Bauer assay. Double disk synergy test (DDST) to confirm metallo-β-lactamase (MBL) production and PCR amplification of specific antibiotic resistance genes was performed. All (100%) 22 P. aeruginosa isolates recovered were resistant to 6 of the β-lactams tested including imipenem but exhibited susceptibility to trimethoprim-sulfamethoxazole. MBL production was observed in 77% of isolates while the most prevalent extended-spectrum β-lactamase (ESBL) genes present included bla(AmpC) (86.9%) followed by bla(TEM) (82.6%). Sulfonamide resistance was largely encoded by sul1 (63.6%) and sul2 (77.3%) genes with a high abundance of class 1 integrons (77.3%) of which 18.2% carried both Intl1 and Intl2. P. aeruginosa found in Sodwana Bay exhibits multi-drug resistance (MDRce) to several pharmaceutically important drugs with the potential to transfer antibiotic resistance to other bacteria if the judicious use of antibiotics for their treatment is not practiced.202235056039
102610.9994Analysis of Wastewater Reveals the Spread of Diverse Extended-Spectrum β-Lactamase-Producing E. coli Strains in uMgungundlovu District, South Africa. Wastewater treatment plants (WWTPs) are major reservoirs of antibiotic-resistant bacteria (ARB), favouring antibiotic resistance genes (ARGs) interchange among bacteria and they can provide valuable information on ARB circulating in a community. This study characterised extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli from the influent and effluent of four WWTPs in uMgungundlovu District, KwaZulu-Natal, South Africa. E. coli was enumerated using the membrane filtration method and confirmed using the API 20E test and real-time polymerase chain reaction. ESBL-producers were phenotypically identified by their susceptibility to the third-generation cephalosporins using the disc diffusion and the double-disc synergy methods against cefotaxime (30 µg) with and without 10 µg clavulanic acid. Genotypic verification was by PCR of the TEM, SHV, and CTX-M genes. The clonality of isolates was assessed by ERIC-PCR. The highest E. coli count ranged between 1.1 × 10(5) (influent) and 4.3 × 10(3) CFU/mL (effluent). Eighty pure isolates were randomly selected, ten from the influent and effluent of each of the four WWTP. ESBLs were phenotypically confirmed in 49% (n = 39) of the isolates, of which 77% (n = 30) were genotypically confirmed. Seventy-three percent of the total isolates were multidrug-resistant (MDR). Only two isolates were susceptible to all antibiotics. Overall, resistance to first and second-generation cephalosporins was higher than to third and fourth generation cephalosporins. Also, 15% of the isolates were resistant to carbapenems. The CTX-M-type ESBL (67%; n = 20) was the most common ESBL antibiotic resistance gene (ARG) followed by TEM (57%; n = 17) and SHV-types (27%; n = 8). Also, a substantial number of isolates simultaneously carried all three ESBL genes. ERIC-PCR revealed a high diversity of isolates. The diversity of the isolates observed in the influent samples suggest the potential circulation of different ESBL-producing strains within the studied district, requiring a more comprehensive epidemiological study to prevent the spread of ESBL-producing bacteria within impoverished communities.202134356780
102720.9994Identification of CTX-M-15 and CTX-M-27 in Antibiotic-Resistant Gram-Negative Bacteria Isolated from Three Rivers Running in Central Italy. The main goal of this study was to identify Gram-negative bacteria resistant to antibiotics, in particular β-lactams, in stream waters and effluents from urban wastewater treatment plants draining into Fino, Tavo, and Saline rivers of the Abruzzo region, Italy. Eight sampling sites were selected because they were the most contaminated by coliforms during previous sampling campaign. One sample for each site was collected for the detection of total and fecal coliforms, Escherichia coli and Enterococcus species by Colilert-18 and Enterolert-E Quanti-Tray/2000. Antibiotic-resistant bacteria, selected on ampicillin and cefotaxime-supplemented agar plates, were identified by EnteroPluri test systems and then confirmed by MALDI-TOF. The resistant determinants were identified and characterized by PCR and sequencing. The microbiological analysis allowed to detect E. coli, total coliforms, fecal coliforms, and enterococci with a coefficient of variation of 215.7%, 212.8%, 242.5%, and 188.5%, respectively. Several Gram-negative bacteria were identified: Serratia liquefaciens, E. coli, Enterobacter cloacae, Citrobacter freundii, Raoultella ornithinolytica, Acinetobacter johnsonii, Aeromonas veronii, Aeromonas hydrophila, and Pseudomonas koreensis. All strains possessed class 1 integrons, insertion sequences, and genes encoding for serin- and metallo-β-lactamases. Extended-spectrum β-lactamases, such as CTX-M-15 and CTX-M-27, were found in Enterobacteriaceae, whereas CphA metallo-β-lactamase was found in A. veronii and A. hydrophila. The main resistance's mechanism to β-lactams observed among the analyzed strains is represented by the production of serin β-lactamases (CTX-M-15, CTX-M-27, and SHV-1) and metallo β-lactamase (CphA).201930994417
106230.9994Prevalence of Biofilm-Forming, ESβLs and Metallo-β-lactamase Producing Gram-Negative MDR Bacteria in the Domestic and Hospital Wastewater of Aligarh City. Gram-negative pathogenic bacteria are a major contributor to antibiotic-resistant infections in hospitals and communities. The emergence of multidrug resistance (MDR) and biofilm formation complicates chemotherapy. This study aimed to assess the prevalence of multidrug-resistant (MDR) biofilm-forming, extended-spectrum beta-lactamase (ESβL) and metallo-beta-lactamase (MβL) producers in wastewater, which pose a public health threat. During 2022-2023, 117 enteric/Gram-negative isolates were isolated using selective culture techniques. Antimicrobial resistance was assessed via disc diffusion assay. ESβL and MβL production was confirmed through phenotypic and PCR-based methods. Biofilm formation was determined using a microtiter plate assay. Biofilms developed on glass coverslips were visualized by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Isolates with bla-(CTX-M), bla-(SHV), and bla-(NDM-1) genes were identified by partial 16S rRNA gene sequencing. A total of 93.16% of isolates were resistant to multiple drug classes (≥ 4), with the increased resistance to ampicillin (100%) and the lowest to gentamicin (12.82%). Biofilm assays indicated that 32.48% of MDR strains formed strong biofilms, 31.62% moderate, and 35.90% weak. β-lactamase activity appeared in 58.97% of isolates, with 56.41% confirmed as ESβL producers. PCR detection in ESβL-positive strains showed 84.61% carried CTX-M, 46.15% SHV, and 53.84% NDM-1 genes. 16S rRNA gene sequencing identified selected MDR strains as Escherichia coli (5), Klebsiella pneumoniae (1), Pseudomonas aeruginosa (1), Salmonella sp. (1), Enterobacter sp. (1), Citrobacter sp. (1), and Proteus sp. (1). The findings reveal the prevalence of biofilm-forming, ESβL, and MβL-producing Gram-negative pathogens in Aligarh's wastewater, underscoring the need for effective treatment to reduce public health risks from MDR bacteria and AMR genes.202540590993
103140.9993Beta-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
102550.9993Detection of Extended Spectrum Beta-Lactamases Resistance Genes among Bacteria Isolated from Selected Drinking Water Distribution Channels in Southwestern Nigeria. Extended Spectrum Beta-Lactamases (ESBL) provide high level resistance to beta-lactam antibiotics among bacteria. In this study, previously described multidrug resistant bacteria from raw, treated, and municipal taps of DWDS from selected dams in southwestern Nigeria were assessed for the presence of ESBL resistance genes which include bla TEM, bla SHV, and bla CTX by PCR amplification. A total of 164 bacteria spread across treated (33), raw (66), and municipal taps (68), belonging to α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Flavobacteriia, Bacilli, and Actinobacteria group, were selected for this study. Among these bacteria, the most commonly observed resistance was for ampicillin and amoxicillin/clavulanic acid (61 isolates). Sixty-one isolates carried at least one of the targeted ESBL genes with bla TEM being the most abundant (50/61) and bla CTX being detected least (3/61). Klebsiella was the most frequently identified genus (18.03%) to harbour ESBL gene followed by Proteus (14.75%). Moreover, combinations of two ESBL genes, bla SHV + bla TEM or bla CTX + bla TEM, were observed in 11 and 1 isolate, respectively. In conclusion, classic bla TEM ESBL gene was present in multiple bacterial strains that were isolated from DWDS sources in Nigeria. These environments may serve as foci exchange of genetic traits in a diversity of Gram-negative bacteria.201627563674
104760.9993Biofilm formation and antibiotic resistance profiles of water-borne pathogens. Water sources (surface water, drinking water, rivers, and ponds) are significant reservoirs for transmitting antibiotic-resistant bacteria. In addition, these waters are an important public health problem because they are suitable environments for transferring antibiotic resistance genes between bacterial species. Our study aimed to assess the prevalence of Extended-spectrum beta-lactamase (ESBL) producing isolates in water samples, the susceptibility of the isolates to the specified antibiotics, the determination of biofilm ability, antibiotic resistance genes, and the molecular typing of the isolates. For this purpose, Polymerase chain reaction (PCR) and Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analyses were used. Out of 70 isolates, 15 (21%) were ESBL producing, and sent for the MALDI-TOF analysis, where Escherichia coli, Acinetobacter calcoaceticus, Enterobacter bugandensis, Acinetobacter pittii, Pseudomonas aeruginosa, Acinetobacter junii, Pseudomonas oleovorans, and Enterobacter ludwigigii were identified. Moreover, colistin resistance genes (mcr 1/2/6, mcr 4, mcr 5, mcr 3/7, and mcr 8), ESBL-encoding genes (bla(SHV), bla(TEM), and bla(CTX-M)) and carbapenemase genes (bla(NDM), bla(OXA-48), and bla(KPC)) using molecular analysis (PCR) were confirmed. The colistin resistance gene was detected at 80% (12/15) in the isolates obtained. The distribution of these isolates according to resistance genes was found as mcr 1/2/6 4 (20%), mcr 3/7 3 (13%), and mcr 5 (40%). Additionally, the isolates harbored bla(SHV)(6.6%) and bla(TEM) (6.6%) genes. However, bla(NDM), bla(OXA-48), bla(KPC), and bla(CTX-M) genes were not detected in any isolates. According to the Congo red agar method, seven (46.6%) isolates showed negative biofilm ability, and eight (53.3%) showed moderate biofilm ability. However, the microplate method detected weak biofilm in 53.3% of the isolates. In conclusion, this study provides evidence for the existence of multidrug-resistant bacteria that co-exist with mcr and ESBL genes in water sources. These bacteria can migrate to other environments and pose increasing threats to public health.202337004897
103870.9993Isolation 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
90280.9993Occurrence of IMP-8, IMP-10, and IMP-13 metallo-β-lactamases located on class 1 integrons and other extended-spectrum β-lactamases in bacterial isolates from Tunisian rivers. BACKGROUND: Antibiotic-resistant bacteria have been surveyed widely in water bodies, but few studies have determined the diversity of antibiotic-resistant bacteria in river waters. This study was undertaken to investigate the origin of resistance among polluted river bacterial isolates in Tunisia. METHODS: In this study 128 isolates resistant to β-lactam antibiotics were obtained from 2 polluted rivers in the north of Tunisia. Isolates were identified using Phoenix phenotyping criteria. The occurrence of bla(TEM), bla(SHV), bla(CTX-M), bla(CMY), bla(VIM), and bla(IMP) was studied by polymerase chain reaction (PCR) amplification and sequencing, and the genetic relatedness of the 16 IMP-producing Klebsiella pneumoniae isolates was analyzed by comparison of XbaI pulsed-field gel electrophoresis (PFGE) profiles. RESULTS: Using Phoenix phenotyping criteria, diverse genera of bacteria were identified with different rates of prevalence and with different minimum inhibitory concentrations against different antibiotics. The occurrence of bla(TEM), bla(SHV), bla(CTXM), bla(CMY), bla(VIM), and bla(IMP) genes was confirmed. The DNA sequences upstream and downstream of bla(IMP) genes were determined, revealing that all IMP-encoding genes constituted the first cassette of class 1 integrons, followed by aacA gene cassettes encoding aminoglycoside resistance. Comparison of PFGE profiles showed that only 2 of the isolates were clonal, the other 14 displaying unique profiles. The bla(CTX-M) gene was the most dominant of the extended-spectrum β-lactamase (ESBL) genes, while the bla(TEM) gene was the second-most dominant. CONCLUSION: The discovery of highly diverse ESBL-producing bacteria and metallo-β-lactamases, particularly bla(IMP), in polluted river water raises alarms with regard to the potential dissemination of antibiotic-resistant bacteria in communities through river environments.201322992193
103390.9993Antimicrobial 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
1039100.9993Genetic Investigation of Beta-Lactam Associated Antibiotic Resistance Among Escherichia Coli Strains Isolated from Water Sources. BACKGROUND: Antimicrobial resistance is an important factor threatening human health. It is widely accepted that antibiotic resistant bacteria such as Escherichia coli (E. coli) released from humans and animals into the water sources, can introduce their resistance genes into the natural bacterial community. OBJECTIVE: The aim of this study was to investigate the prevalence of bla(TEM), bla(CTX), bla(SHV), bla(OXA) and bla(VEB) associated-antibiotic resistance among E. coli bacteria isolated from different water resources in Iran. METHODS: The study contained all E. coli strains segregated from different surface water sources. The Kirby-Bauer method and combined discs method was determined in this study for testing antimicrobial susceptibility and strains that produced Extended-Spectrum Beta Lactamases (ESBL), respectively. DNA extraction kit was applied for genomic and plasmid DNA derivation. Finally the frequency of resistant genes including bla(TEM), bla(CTX), bla(SHV), bla(OXA) and bla(VEB) in ESBL producing isolates were studied by PCR. RESULTS: One hundred E. coli strains were isolated and entered in the study. The highest antibiotic resistance was observed on clindamycin (96%). Moreover, 38.5% isolates were ESBL producers. The frequency of different ESBLs genes were 37%, 27%, 27%, and 25% for bla(TEM), bla(CTX), bla(SHV), and bla(OXA) , respectively. The bla(VEB) wasn't found in any isolates. CONCLUSION: The study revealed a high prevalence of CTX-M, TEM, SHV and OXA genes among E. coli strains in surface water resources. In conclusion, these results raised a concern regarding the presence and distribution of these threatening factors in surface water sources and its subsequent outcomes.201729151997
1034110.9993Detection 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
1053120.9993Antimicrobial Resistance and Extended-Spectrum Beta-Lactamase Genes in Enterobacterales, Pseudomonas and Acinetobacter Isolates from the Uterus of Healthy Mares. Antibiotic-resistant bacteria are a growing concern for human and animal health. The objective of this study was to determine the antimicrobial resistance and extended-spectrum beta-lactamase genes in Enterobacterales, Pseudomonas spp. and Acinetobacter spp. isolates from the uterus of healthy mares. For this purpose, 21 mares were swabbed for samples, which were later seeded on blood agar and MacConkey agar. The isolates were identified using MALDI-TOF and the antimicrobial susceptibility test was performed using the Kirby-Bauer technique. To characterize the resistance genes, a polymerase chain reaction (PCR) scheme was performed. Of the isolates identified as Gram-negative, 68.8% were Enterobacterales, represented by E. coli, Enterobacter cloacae, Citrobacter spp., and Klebsiella pneumoniae; 28.1% belonged to the genus Acinetobacter spp.; and 3.1% to Pseudomonas aeruginosa. A 9.3% of the isolates were multidrug-resistant (MDR), presenting resistance to antibiotics from three different classes, while 18.8% presented resistance to two or more classes of different antibiotics. The diversity of three genes that code for ESBL (bla(TEM), bla(CTX-M) and bla(SHV)) was detected in 12.5% of the strains. The most frequent was bla(SHV), while bla(TEM) and bla(CTX-M) were present in Citrobacter spp. and Klebsiella pneumoniae. These results are an alarm call for veterinarians and their environment and suggest taking measures to prevent the spread of these microorganisms.202337764953
1058130.9993First 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
1150140.9993Occurrence of multidrug resistance associated with extended-spectrum β‑lactamase and the biofilm forming ability of Escherichia coli in environmental swine husbandry. Extended-spectrum beta-lactamase (ESBL) production and biofilm formation are mechanisms employed by Escherichia coli to resist beta-lactam antibiotics. Thus, we aimed to examine antibiotic resistance associated with ESBL production and biofilm formation in E. coli isolates from swine farms in Southern Thailand. In total, 159 E. coli isolates were obtained, with 44 isolates identified as ESBL producers, originating from feces (18.87 %) and wastewater (8.80 %) samples. All ESBL-producing strains exhibited resistance to ampicillin (100 %), followed by the cephalosporin group (97.73 %) and tetracycline (84.09 %). Multidrug resistance was observed in 17 isolates (38.63 %). Among the isolates from feces samples, the bla(GES) gene was the most prevalent, detected in 90 % of the samples, followed by bla(CTX-M9) (86.67 %) and bla(CTX-M1) (66.67 %), respectively. In the bacteria isolated from wastewater, both bla(GES) and bla(CTX-M9) genes were the predominant resistance genes, detected in 100 % of the isolates, followed by bla(CTX-M1) (64.29 %) and bla(TEM) (50 %), respectively. Strong biofilm formation was observed in 11 isolates (36.67 %) from feces and 4 isolates (25.57 %) from wastewater samples. Notably, nearly 100 % of ESBL-producing strains isolated from feces tested positive for both pgaA and pgaC genes, which play a role in intracellular adhesion and biofilm production. These findings contribute to the understanding and potential control of ESBL-producing E. coli, and the dissemination of antibiotic resistance and biofilm-related genes in swine farms.202337976973
1165150.9993Isolation, Antimicrobial Susceptibility Profile and Detection of Sul1, blaTEM, and blaSHV in Amoxicillin-Clavulanate-Resistant Bacteria Isolated From Retail Sausages in Kampar, Malaysia. BACKGROUND: Due to the overuse of antibiotics in livestock as a growth-promoting agent, the emergence of multi-antibiotic resistant bacteria is becoming a concern. OBJECTIVES: In this study, we aimed to detect the presence and discover the molecular determinants of foodborne bacteria in retail sausages resistant towards the antibacterial agent amoxicillin-clavulanate. METHODS: Two grams of sausages were chopped into small pieces and transferred into sterile Luria-Bertani (LB) enrichment broths overnight before they were plated on MacConkey agar petri dishes. The bacteria isolated were then screened for amoxicillin-clavulanate resistance, and an antimicrobial susceptibility test of each isolate was performed by using the disc diffusion method. Double synergy and phenotypic tests were carried out to detect the presence of extended spectrum β-lactamase (ESBL). API 20E kit was used to identify the Enterobacteriaceae. All isolates were further examined by polymerase chain reaction (PCR) for resistant genes blaOXA-1, blaOXA-10, plasmid-mediated AmpC (blaCMY and blaDHA), and the chromosome-mediated AmpC, Sul1, blaTEM, and blaSHV genes. RESULTS: A total of 18 amoxicillin-clavulanate resistant isolates were obtained from seven different types of retail sausages. Only half of them were identified as Enterobacteriaceae, but none were ESBL-producers. All the 18 isolated strains demonstrated resistance towards amoxicillin-clavulanate, penicillin and oxacillin (100%), cefotaxime (71.4%), cefpodoxime (66.7%), and ampicillin (83.3%). blaTEM was the most frequently detected β-lactamase gene. Both plasmid- and chromosomal-bound blaTEM genes were detected in all of the isolated Enterobacteriaceae. blaSHV and Sul1 accounted for 22.2% and 11.1% of the amoxicillin-clavulanate resistant isolates, respectively, whereas blaAMPC, blaCMY, blaDHA, blaOXA-1, and blaOXA-10 were not found in any of the isolates. The only one ESBL-producing bacteria detected in this study was Chryseobacterium meningosepticum, which harbored the blaTEM gene. CONCLUSIONS: The multidrug resistant bacteria that carry antibiotic resistant genes from retail sausages may increase the risk of transmission to humans via the consumption of contaminated sausages. Stricter measures must be taken to address the use of antibiotics in animal agriculture and to consider their potential impact on human health.201627942365
1445160.9993Rapid Detection of Beta-Lactamases Genes among Enterobacterales in Urine Samples by Using Real-Time PCR. The objective of this study was to develop and evaluate newly improved, rapid, and reliable strategies based on real-time PCR to detect the most frequent beta-lactamase genes recorded in clinical Enterobacterales strains, particularly in Tunisia (bla(SHV12) , bla(TEM) , bla(CTX-M-15) , bla(CTX-M-9) , bla(CMY-2) , bla(OXA-48) , bla(NDM-1) , and bla(IMP) ) directly from the urine. Following the design of primers for a specific gene pool and their validation, a series of real-time PCR reactions were performed to detect these genes in 78 urine samples showing high antibiotic resistance after culture and susceptibility testing. Assays were applied to DNA extracted from cultured bacteria and collected urine. qPCR results were compared for phenotypic sensitivity. qPCR results were similar regardless of whether cultures or urine were collected, with 100% sensitivity and specificity. Out of 78 multiresistant uropathogenic, strains of Enterobacterales (44 E. coli and 34 K. pneumoniae strains) show the presence of the genes of the bla group. In all, 44% E. coli and 36 of K. pneumoniae clinical strains harbored the bla group genes with 36.4%, 52.3%, 70.5%, 68.2%, 18.2%, and 4.5% of E. coli having bla(SHV-12) , bla(TEM) , bla(CTX-M 15) , bla(CTX-M-9) , bla(CMY-2) , and bla(OXA-48) group genes, respectively, whereas 52.9%, 67.6%, 76.5%, 35.5%, 61.8, 14.7, and 1.28% of K. pneumoniae had bla(SHV-12) , bla(TEM) , bla(CTX-M 15) , bla(CTX-M-9) , bla(CMY-2) , bla(OXA-48) , and bla(NDM-1) group genes, respectively. The time required to have a result was 3 hours by real-time PCR and 2 to 3 days by the conventional method. Resistance genes of Gram-negative bacteria in urine, as well as cultured bacteria, were rapidly detected using qPCR techniques. These techniques will be used as rapid and cost-effective methods in the laboratory. Therefore, this test could be a good candidate to create real-time PCR kits for the detection of resistance genes directly from urine in clinical or epidemiological settings.202235978630
1162170.9993Abundance of extended-spectrum β-lactamase genes among intestinal Escherichia coli strains from drug users. Drug users may represent a hidden reservoir of antibiotic resistance genes among their intestinal flora due to the poor hygiene and inappropriate use of antibiotics. Therefore, this study was focused to examine the prevalence of extended-spectrum β-lactamase (ESBL) genes among intestinal Escherichia coli isolated from drug users in Ahvaz, Iran. Among clients of toxicology laboratory who were confirmed their addiction to each of Morphine, Amphetamine or Methamphetamine, 109 drug users were examined voluntarily for infection with hepatitis B or C using commercial enzyme linked immunosorbent assays (ELISA) method. Their stool specimens were obtained to isolate intestinal E. coli. The disc diffusion and combination disk methods were conducted to demonstrate antibiotic resistance pattern and phenotypically ESBL producers. ESBL-encoding genes (bla-TEM, bla-CTX-M, and bla-SHV) were also examined by PCR. Based on results, hepatitis C infection was more prevalent than hepatitis B among drug users. Of 109 isolates, a total of 57 (52.29%) ESBL positive E. coli were obtained from drug users and bla-TEM gene (60.55%) was found to be the most prevalent type, followed by bla-CTX-M (40.36%) and bla-SHV (39.44%). All isolates represented different resistance levels to tested antibiotics and 54.43% of the ESBL‑producing isolates showed multidrug resistance (MDR) and the most frequent MDR pattern was simultaneous resistance to the seven (27.90%) of antimicrobials particularly erythromycin, penicillin, amoxycilin, cefteriaxon, cefotaxim, tetracycline and trimethoprim-Sulfamethoxazole. Fecal carriage of ESBL-production and MDR commensal isolates such as E. coli among drug users underlines the risk of transferring resistance genes between nonpathogenic and pathogenic bacteria.202133837441
1127180.9993Extended spectrum beta-lactamase and aminoglycoside modifying enzyme genes in multi drug resistant Gram-negative bacteria: A snapshot from a tertiary care centre. BACKGROUND: This study aims to enhance the existing knowledge of the prevalence of genes responsible for beta-lactam resistance and aminoglycoside resistance in gram negative organisms by molecular detection of extended spectrum beta-lactamase and aminoglycoside modifying enzymes in multidrug-resistant gram-negative bacteria. METHODS: Out of 864 gram-negative isolates, 710 were phenotypically identified as multidrug-resistant by antibiotic susceptibility testing. From the above isolates, 102 representative isolates as per sample size calculated were selected for further molecular studies. The presence of blaTEM, blaCTX-M blaSHV, and five AmpC genes was detected by real-time polymerase chain reaction (PCR). Conventional PCR was performed to detect seven aminoglycoside modifying enzyme genes namely aac(6')-Ib, aac(6')-Ic, aac(3)-Ia, aac(3)-Ib, aac(3)-IIa, ant(2'')-Ia, and ant(4'')-IIa. RESULTS: Most common multidrug-resistant isolate was Klebsiella pneumoniae (35%) followed by Escherichia coli (30%). Among the 102 selected isolates all harboured blaTEM gene, 71 (69.6%) harboured blaCTX-M gene and 48 (47%) blaSHV gene. Among the selected isolates 60% showed the presence of AmpC genes. Most common aminoglycosie modifying enzyme gene was AAC 6' Ib (51%) followed by ANT 2" Ia (36%). CONCLUSION: This study suggests a wider use of molecular methods using specific PCR amplification of resistance genes. It would be beneficial to perform the molecular identification of antimicrobial resistance genes to effectively monitor and manage antibiotic resistance, administer appropriate antimicrobial medication, practice antimicrobial stewardship and improve hospital infection control procedures.202439734850
1124190.9993Molecular Identification of Extended-Spectrum β-lactamase and Integron Genes in Klebsiella Pneumonia. INTRODUCTION: Infections caused by Gram negative bacteria, producing extended-spectrum β-lactamase, including Klebsiella pneumoniae are increasing all over the world with high morbidity and mortality. The aim of the present study was determined antimicrobial profile susceptibility and the prevalence of antibiotic resistance genes by multiplex PCR. METHODS: In the present study, we obtained one-hundred isolates of K. pneumoniae from different clinical samples. The antibiotic susceptibility testing was done in thirteen antibiotic and, therefore, M-PCRs were conducted using the DNA amplification for detection of ESBLs (blaTEM, blaCTX-M, blaSHV) and int (I, II, III) genes. RESULTS: The results of resistance to amoxicillin/clavulanate, ciprofloxacin, amikacin, trimethoprim-sulfamethoxazole, cefotaxime, ampicillin, aztreonam, imipenem, gentamicin, ceftazidime, Cefepime, ceftriaxone and levofloxacin were obtained 37%, 37%, 93%, 84%, 52%, 87%, 59%, 8%, 24%, 67%, 52%, 43% and 26%, respectively. The frequency of the extended-spectrum β-lactamase K. pneumoniae was obtained 37%. The prevalence of resistance genes of ESBLs in the M-PCR method showed that the blaTEM, blaCTX and blaSHV were 38%, 24% and 19%, respectively, however, only 8 (8%) out of 100 isolates were found to have positive outcomes for the existence of class 1 integrons and there were no detected class 2 or class 3 integrons. CONCLUSIONS: Our results recommend the likely co-carriage of some ESBLs genes and antibiotic resistance integrons on the same plasmids harboring the MDR genes.201627935927