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Klebsiellapneumoniae is the most important species of the Klebsiella genus and often causes hospital infections. These bacteria have a high resistance to most of the available drugs, which has caused concern all over the world. In this study, we investigated the antibiotic resistance profile and the ability to produce extended-spectrum beta-lactamase (ESBL) among K. pneumoniae isolates, and then we investigated the relationship between these two factors with biofilm formation and the prevalence of different virulence genes. In this study, 130 isolates of K. pneumoniae isolated from wounds were investigated. The antibiotic resistance of the isolates was evaluated by the disk diffusion method. The microtiter plate method was used to measure biofilm formation. The prevalence of virulence genes was detected by multiplex PCR. Among the examined isolates, 85.3% showed multidrug resistance. 87.6% of the isolates were ESBL-positive. Imipenem, meropenem, and fosfomycin were the most effective drugs. The ability of the isolates to produce biofilm was strong (80%), moderate (12.3%), and weak (7.6%), respectively. fimH, mrKD, entB, and tolC virulence genes were observed in all isolates. High prevalence of antibiotic resistance (especially multidrug resistance), high prevalence of ESBL-producing isolates, the ability of all isolates to biofilm formation, and the presence of fimH, mrKD, entB, and tolC virulence genes in all isolates show the importance of these factors in the pathogenesis of K. pneumoniae isolates in Iraq. [ABSTRACT FROM AUTHOR]

Background/Objectives: Over the past few decades, extended-spectrum β-lactamase (ESBL)-producing bacteria have become a great concern in healthcare systems worldwide, imposing large burdens by increasing antimicrobial resistance and patient morbidity. Given the high mortality rates and emergence of multidrug-resistant (MDR) strains, monitoring ESBL prevalence and resistance patterns is crucial. This study aimed to evaluate ESBL-producing Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae over three years, focusing on phenotypic distribution and resistance profiles. Methods: A total of 1599 ESBL-producing bacterial samples were collected and analysed. A panel of 20 antibiotics was tested to determine resistance traits. Data were recorded on phenotypical distribution, isolation types, changes in antibiotic resistance, and the relation of such changes to antibiotic consumption (defined daily dose) from clinical isolates. Results: Phenotypical analysis revealed the minimal presence of the Cefotaximase from Munich (CTX-M) phenotype in E. coli and K. pneumoniae, creating a distinct epidemiological profile compared to global patterns. Shifts in isolation trends, particularly in P. mirabilis, suggest an expected increase in associated-mortality-rate in the coming years. While resistance trends were not statistically significant, MDR and extensively drug-resistant (XDR) strains were identified across all three bacteria. Only meropenem showed consistent 100% efficacy against E. coli, with other antibiotics displaying only partial effectiveness. Conclusions: These findings highlight the need for ongoing surveillance of ESBL-producing bacteria and underscore challenges in managing antibiotic resistance due to limited efficacy of last-resort treatments. The unique phenotypical distribution observed could impact local resistance management strategies in hospital settings in the coming years. [ABSTRACT FROM AUTHOR]

E. coli and total coliforms are the most widely used indicator organisms for microbial monitoring of drinking water and recreational freshwater. In many remote and low-resource settings, however, conventional laboratory methods for quantifying these indicators are challenging or infeasible to perform due to limited access to laboratory facilities. The availability of rapid, low-cost methods for quantifying indicator organisms in freshwater samples without the need for laboratory facilities is crucial to facilitate the rapid and robust monitoring of microbial water quality in these types of settings. The global misuse and abuse of antimicrobials have contributed to the rise of antimicrobial resistance. Thus, simple culture methods are needed to detect indicators of such bacteria in freshwaters. In 2021, the World Health Organization released the Tricycle protocol to address this issue by providing guidance for culture-based detection of extended-spectrum beta-lactamase (ESBL)-producing E. coli in environmental samples. Our research goal was to compare the 100-ml sample volume ESBL E. coli quantal and enumerative commercial tests against the more complex Tricycle protocol to detect and quantify ESBL E. coli in surface waters. Both commercial tests gave results comparable with the results obtained using the Tricycle protocol, and the quantal and enumerative commercial tests were easier and faster to perform than the Tricycle protocol. [ABSTRACT FROM AUTHOR]

Specific data concerning the efficacy of alternative antibiotics for carbapenems against complicated urinary tract infections (cUTIs) attributed to antimicrobial-resistant (AMR) uropathogens are lacking. This study aimed to assess the efficacy of carbapenems and non-carbapenem antibiotics in the clinical outcomes of cUTIs caused by AMR uropathogens. In this systematic review and meta-analysis, databases, including MEDLINE/PubMed, the Cochrane Library, Embase and ClinicalTrials.gov , were searched. The study eligibility criteria were research articles conducted as randomised controlled trials that evaluated the composite outcomes of cUTIs. Participants were adult patients with cUTIs caused by gram-negative uropathogens resistant to third-generation cephalosporins. The intervention involved a non-carbapenem class of antimicrobial agents with in vitro activities against gram-negative uropathogens resistant to third-generation cephalosporins. Two independent researchers assessed the risk-of-bias using the second version of the Cochrane risk-of-bias tool for randomised trials. The treatment effects on each outcome were estimated as a risk ratio (RR) with a 95 % confidence interval (CI) using the random-effects model. Heterogeneity was assessed using the Cochrane Q-test and I2 statistics. Through database searches, 955 articles were retrieved. After screening the titles and abstracts, 52 articles were screened in full text. Finally, 12 studies met the inclusion criteria. No significant differences in efficacy were observed between alternative antibiotics and carbapenems (composite outcome, RR, 0.96; 95 % CI, 0.63–1.49; I2 = 21 %; low certainty of evidence). Alternative antibiotics had clinical efficacy similar to that of carbapenems for treating patients with cUTI caused by gram-negative uropathogens resistant to third-generation cephalosporins. [ABSTRACT FROM AUTHOR]

Background: In Gabon, studies on the characterization of extended-spectrum beta-lactamase-producing Escherichia coli in young children with diarrhoea are almost nonexistent. The objective was to evaluate the prevalence of antibiotic resistance to extended-spectrum beta-lactamase-producing Escherichia coli in children at public hospitals in Franceville, Gabon. Methods: Seventy diarrhoea faecal samples were collected from children aged 0–5 years. The culture and isolation of colonies were carried out on MacConkey agar. The colonies were identified using VITEK 2. The determination of the extended-spectrum beta-lactamase's profiles was accomplished using the double disk method. The identification of phylogroups and pathotypes was performed by PCR. Identification of the ESBL genes was performed by sequencing. Results: A total of 26 strains of Escherichia coli (33.0%) were identified from 78 bacterial isolates. Twenty (77.0%) Escherichia coli strains carried extended-spectrum beta-lactamases blaCTX-M-15 and 5.0% carried blaSHV-12 subtypes. Phylogroup D (62.0%) was predominant, followed by B1 (12.0%), B2 (8.0%) and E (4.0%). The bacterial pathogens causing diarrhoea were enterohemorrhagic E. coli (12.0%), typical enteropathogenic Escherichia coli (8.0%), atypical enteropathogenic Escherichia coli (4.0%), Enteroaggregative Escherichia coli (4.0%) and enteroinvasive E. coli (4.0%). Conclusions: This study showed a high prevalence of extended-spectrum beta-lactamase, Escherichia coli of phylogroup D and pathotype enterohemorrhagic Escherichia coli in children under 5 years old in public hospitals in Franceville, most probably due to the misuse or inappropriate consumption of beta-lactams. [ABSTRACT FROM AUTHOR]

Abstract Background The worldwide prevalence of multi-drug resistance (MDR) in Gram-negative bacteria (GNB), particularly related to extended-spectrum beta-lactamases (ESBLs) and carbapenemases, poses significant global public health and clinical challenges. Objectives To characterize ESBL-producing Gram-negative bacilli, within a pediatric hospital in Gaza using whole genome sequencing (WGS). Methods A total of 158 clinical isolates of Gram-negative bacilli were collected from Al-Nasser Pediatric Hospital. These isolates were tested for ESBL production using the double disk synergy test. The antibiotic susceptibility profile was determined using the Kirby Bauer method following the Clinical and Laboratory Standard Institute guidelines. Selected 15 phenotypically MDR isolates were whole-genome sequenced and characterized for their genome-based species identity and antibiotic resistance gene profile. Results Of the 158 isolates, 93 (58.9%) were positive for ESBL production. The frequency of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, and Serratia marcescens was 50%, 22.7%, 22.7%, 1.8%, 1.2%, and 1.2% respectively. The prevalence of ESBL among urine, pus, blood, and sputum was 64%, 44%, 23%, and 63.6%, respectively. Chloramphenicol, Imipenem, and Meropenem were the most effective antibiotics against ESBL producers. In sequenced isolates, an average of six anti-microbial resistance (AMR) genes were noted per isolate, where one of them carried up to 13 antibiotic resistance genes. Carbapenem resistance genes such as blaKPC-2(6.6%), blaPDC-36/12 (6.6%), and blaPOM-1 (6.6%) were detected. All the sequenced E. coli isolates (n = 8) showed multiple resistance genes, mainly against β-lactamase (25.0%), aminoglycosides (37.5%), sulfonamides (37.5%), and genes conferring resistance to tetracyclines (25.0). Conclusion Our results showed a high prevalence of ESBL-producing GNB isolated from a pediatric hospital in the Gaza Strip. Various antibiotic resistance genes were identified, including those encoding ESBL and carbapenems. The results highlight the significant challenge posed by MDR in GNB and emphasize the need for effective antibiotic strategies. Given the high endemicity observed in various studies from Palestine, it is important to conduct clinical and molecular epidemiology research to identify risk factors, transmission patterns, and clinical outcomes associated with GNB strains that carry ESBL and carbapenem resistance genes.

Abstract Background Extended-spectrum β-lactamase -producing Enterobacterales (ESBL-E) are important zoonotic pathogens that can cause serious clinical infections, also in horses. Preventing the spread of ESBL-E, especially in the equine hospital environment, is key to reducing the number of difficult-to-treat infections. Estimating the local prevalence of ESBL-E in horses is crucial to establish targeted infection control programs at equine hospitals. We conducted a prevalence and risk factor study in equine patients on admission to an equine teaching hospital in Finland through a rectal ESBL-E screening specimen of the horse and a questionnaire. Results The prevalence of ESBL-E in admitted horses was 3% (5/161, 95% CI 1–7%); none of the tested factors remained statistically significant in multivariate analysis, although antimicrobial treatment within three months was borderline significant (p = 0.052). Extended-spectrum β-lactamase -producing Klebsiella pneumoniae ST6179:CTX-M-15 was detected in three horses using whole-genome sequencing, which in combination with patient records suggested nosocomial transmission. Escherichia coli isolates were ST1250:CTX-M-1 (n = 1), ST1079:CTX-M-1 (n = 1), and ST1245:CTX-M-14 (n = 1). Multiple virulence genes were detected in the ESBL-E isolates. In the ESBL-E positive horses enrolled in a one-year follow-up study, ESBL-E were unlikely to be isolated in rectal screening specimens after the initial positive specimen. Conclusions The prevalence of ESBL-E in horses visiting a veterinary teaching hospital in Finland is low, indicating an overall low prevalence estimate in the country’s equine population. No statistically significant risk factors were identified, likely due to the low number of cases. The duration of ESBL-E carriage is likely to be very short in horses.

Background: The worldwide prevalence of multi-drug resistance (MDR) in Gram-negative bacteria (GNB), particularly related to extended-spectrum beta-lactamases (ESBLs) and carbapenemases, poses significant global public health and clinical challenges. Objectives: To characterize ESBL-producing Gram-negative bacilli, within a pediatric hospital in Gaza using whole genome sequencing (WGS). Methods: A total of 158 clinical isolates of Gram-negative bacilli were collected from Al-Nasser Pediatric Hospital. These isolates were tested for ESBL production using the double disk synergy test. The antibiotic susceptibility profile was determined using the Kirby Bauer method following the Clinical and Laboratory Standard Institute guidelines. Selected 15 phenotypically MDR isolates were whole-genome sequenced and characterized for their genome-based species identity and antibiotic resistance gene profile. Results: Of the 158 isolates, 93 (58.9%) were positive for ESBL production. The frequency of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, and Serratia marcescens was 50%, 22.7%, 22.7%, 1.8%, 1.2%, and 1.2% respectively. The prevalence of ESBL among urine, pus, blood, and sputum was 64%, 44%, 23%, and 63.6%, respectively. Chloramphenicol, Imipenem, and Meropenem were the most effective antibiotics against ESBL producers. In sequenced isolates, an average of six anti-microbial resistance (AMR) genes were noted per isolate, where one of them carried up to 13 antibiotic resistance genes. Carbapenem resistance genes such as blaKPC-2(6.6%), blaPDC-36/12 (6.6%), and blaPOM-1 (6.6%) were detected. All the sequenced E. coli isolates (n = 8) showed multiple resistance genes, mainly against β-lactamase (25.0%), aminoglycosides (37.5%), sulfonamides (37.5%), and genes conferring resistance to tetracyclines (25.0). Conclusion: Our results showed a high prevalence of ESBL-producing GNB isolated from a pediatric hospital in the Gaza Strip. Various antibiotic resistance genes were identified, including those encoding ESBL and carbapenems. The results highlight the significant challenge posed by MDR in GNB and emphasize the need for effective antibiotic strategies. Given the high endemicity observed in various studies from Palestine, it is important to conduct clinical and molecular epidemiology research to identify risk factors, transmission patterns, and clinical outcomes associated with GNB strains that carry ESBL and carbapenem resistance genes. [ABSTRACT FROM AUTHOR]

Background There are limited and conflicting data regarding the impact of race or ethnicity on the rate of gram-negative antimicrobial resistance. This study was performed to determine whether there is a difference in extended-spectrum beta-lactamase (ESBL) Escherichia coli infection or colonization in minoritized patients when compared to White patients from a diverse US Midwestern city. Methods A case control study was performed, with controls with non-ESBL E. coli matched 1:1 to patients with ESBL-producing E coli based on age, sex, and ZIP code. A variety of other evidence-based factors for ESBL Enterobacterales infection and colonization were collected via chart review. Multivariate conditional logistic regression assessed the odds of minoritized patients as compared to White patients, while controlling for other common risk factors for ESBL Enterobacterales. Results A total of 364 matched pairs were included in the analysis. Females were the majority of the sample (91%), with median age of 65 years. The majority of the sample identified as White (73%), followed by Hispanic (14%) and Black (10%). Urine cultures made up the majority of the cultures in the sample (97%), and this was similar between ESBL and non-ESBL groups. While controlling for these risk factors for ESBL E coli , minoritized patients had a statistically significant greater odds of ESBL-producing E coli (odds ratio, 2.53; 95% confidence interval, 1.68–3.82). Conclusions In our sample, which is demographically similar to the United States, minoritized patients had higher odds of ESBL-producing E coli. Further research on the drivers for this disparity is needed. [ABSTRACT FROM AUTHOR]

Background: Extended-spectrum β-lactamase -producing Enterobacterales (ESBL-E) are important zoonotic pathogens that can cause serious clinical infections, also in horses. Preventing the spread of ESBL-E, especially in the equine hospital environment, is key to reducing the number of difficult-to-treat infections. Estimating the local prevalence of ESBL-E in horses is crucial to establish targeted infection control programs at equine hospitals. We conducted a prevalence and risk factor study in equine patients on admission to an equine teaching hospital in Finland through a rectal ESBL-E screening specimen of the horse and a questionnaire. Results: The prevalence of ESBL-E in admitted horses was 3% (5/161, 95% CI 1–7%); none of the tested factors remained statistically significant in multivariate analysis, although antimicrobial treatment within three months was borderline significant (p = 0.052). Extended-spectrum β-lactamase -producing Klebsiella pneumoniae ST6179:CTX-M-15 was detected in three horses using whole-genome sequencing, which in combination with patient records suggested nosocomial transmission. Escherichia coli isolates were ST1250:CTX-M-1 (n = 1), ST1079:CTX-M-1 (n = 1), and ST1245:CTX-M-14 (n = 1). Multiple virulence genes were detected in the ESBL-E isolates. In the ESBL-E positive horses enrolled in a one-year follow-up study, ESBL-E were unlikely to be isolated in rectal screening specimens after the initial positive specimen. Conclusions: The prevalence of ESBL-E in horses visiting a veterinary teaching hospital in Finland is low, indicating an overall low prevalence estimate in the country's equine population. No statistically significant risk factors were identified, likely due to the low number of cases. The duration of ESBL-E carriage is likely to be very short in horses. [ABSTRACT FROM AUTHOR]

Foodborne pathogens' transmission is essential in the spread of antibiotic resistance, and extended-spectrum beta-lactamase-producing Escherichia coli especially threatens public health. E. coli plays an essential role in the resistance to commonly used beta-lactam group antibiotics. Ready-to-eat (RTE) stuffed mussels are among many restaurants and street vendors, presenting potential health risks of food hygiene origin. 200 RTE stuffed mussels were collected from the Asian and European sides of Istanbul and analysed for the presence of E. coli. As a result of PCR analysis, E. coli was detected in 7 (3.5%) samples. An antibiotic susceptibility test was performed using the disc diffusion method to determine ESBL and carbapenem resistance. All isolates were resistant to ampicillin. The double-disk synergy test was performed as an ESBL phenotypic confirmation test, and no phenotypically ESBL-producing E. coli were detected. The bla TEM gene was detected in one isolate (14.2%) by mPCR, but bla CTX-M, bla SHV, and bla OXA genes were not observed. Meropenem and imipenem were used with the disk diffusion method for carbapenem resistance study, and no resistant isolate was found. Carbapenem resistance genes were investigated by monoplex PCR, and bla NDM-1, bla OXA-48, bla VIM, and bla IMP resistance genes were not detected. This is the first report on ESBL-producing E. coli in RTE stuffed mussels in Türkiye, which draws attention to a public health risk. [ABSTRACT FROM AUTHOR]

Background: Veno-venous extracorporeal membrane oxygenation (V-V ECMO) is a rapidly expanding life-support technique worldwide. The most common indications are severe hypoxemia and/or hypercapnia, unresponsive to conventional treatments, primarily in cases of acute respiratory distress syndrome. Concerning potential contraindications, there is no mention of microbiological history, especially related to multi-drug resistant (MDR) bacteria isolated before V-V ECMO placement. Our study aims to investigate: (i) the prevalence and incidence of MDR Gram-negative (GN) bacteria in a cohort of V-V ECMOs; (ii) the risk of 1-year mortality, especially in the case of predetected MDR GN bacteria; and (iii) the impact of annual hospital V-V ECMO volume on the probability of acquiring MDR GN bacteria. Methods: All consecutive adults admitted to the Intensive Care Units of 5 Italian university-affiliated hospitals and requiring V-V ECMO were screened. Exclusion criteria were age < 18 years, pregnancy, veno-arterial or mixed ECMO-configuration, incomplete records, survival < 24 h after V-V ECMO. A standard protocol of microbiological surveillance was applied and MDR profiles were identified using in vitro susceptibility tests. Cox-proportional hazards models were applied for investigating mortality. Results: Two hundred and seventy-nine V-V ECMO patients (72% male) were enrolled. The overall MDR GN bacteria percentage was 50%: 21% (n.59) detected before and 29% (n.80) after V-V ECMO placement. The overall 1-year mortality was 42%, with a higher risk observed in predetected patients (aHR 2.14 [1.33–3.47], p value 0.002), while not in 'V-V ECMO-acquired MDR GN bacteria' group (aHR 1.51 [0.94–2.42], p value 0.090), as compared to 'non-MDR GN bacteria' group (reference). Same findings were found considering only infections. A larger annual hospital V-V ECMO volume was associated with a lower probability of acquiring MDR GN bacteria during V-V ECMO course (aOR 0.91 [0.86–0.97], p value 0.002). Conclusions: 21% of MDR GN bacteria were detected before; while 29% after V-V ECMO connection. A history of MDR GN bacteria, isolated before V-V ECMO, was an independent risk factor for mortality. The annual hospital V-V ECMO volume affected the probability of acquiring MDR GN bacteria. Trial Registration ClinicalTrial.gov Registration Number NCTNCT06199141, date 12.26.2023. [ABSTRACT FROM AUTHOR]

Background and Aims: The role of fomites in the transmission of infectious diseases is becoming more important because of the possibility that contaminated surfaces act as reservoirs of diseases. The aim of this study was to ascertain the level of bacterial contamination and the prevalence of antibiotic-resistant genes on the most touched surfaces on public transport. Methods: One hundred samples were collected from door handles and armrests of buses and tricycles, respectively at Isale-Oko motor garage, Sagamu. Bacteria were isolated from the samples and identified following standard microbiological techniques. Antibiotic susceptibility testing was done with the Kirby-Bauer disc diffusion method. The presence of antibiotic-resistant genesvanA, drfA, dfrG genes, and extended-spectrum betta-lactamase (ESBL) genes in Gram-negatives were screened by polymerase chain reaction (PCR) method. Results: Out of the samples tested, 91% were positive for bacterial contamination. Among the 91 positive samples, 126 bacteria were identified, comprising 98 Gram-positive and 28 Gram-negative bacterial isolates. Staphylococcus aureus had the highest overall frequency of occurrence with 62 (49.2%) isolates. Among the Gram-positives, azithromycin resistance was present in 35(56.5%) S. aureus and 19(52.8%) Staphylococcus epidermidis. Salmonella species was the most resistant to ciprofloxacin (100%). dfrG was the most detected among trimethoprim-resistant genes occurring in 11(55%) of multidrug-resistant S. aureus and 6(54.6%) of S. epidermidis. vanA gene was present in S. aureus (20%). dfrA was present in only Klebsiella pneumoniae and Esherichia coli. E. coli and Shigella species carried blaTEM while blaSHV was found in Pseudomonas aeruginosa. Conclusion: The most-touched surfaces of public transportation can serve as a substantial source of spread for potentially harmful bacteria. [ABSTRACT FROM AUTHOR]

Proteus species are rod-shaped, Gram-negative bacteria that cause opportunistic infections in the urinary tract and occasionally in the gastrointestinal tract. They are implicated in infections like cystitis and pyelonephritis, particularly in immunocompromised individuals, and are frequently present in cases of asymptomatic bacteriuria. Herein, we aimed to investigate the co-occurrence of extended-spectrum beta-lactamase (ESBL) enzyme production and multidrug resistance (MDR) among Proteus spp. Isolated from patients attending selected hospitals in Northeastern Nigeria. A total of 1,500 clinical samples from consenting patients across six states in the Northeastern region of Nigeria were collected. The samples were cultured on Blood agar, and growth resembling that of Proteus species were again subcultured onto MacConkey agar to obtain discrete colonies, further confirmed using biochemical tests. Antibiotics susceptibility test was carried out for all isolates using the Kirby-Bauer disc diffusion method, coupled with a screening of the production of extended-spectrum beta-lactamase using the Combined Disc Diffusion Method. Of the 1500 samples collected, 144 yielded positive growth for Proteus spp., resulting in a prevalence rate of 9.60%. Among these Proteus isolates, three species were identified, with Proteus mirabilis (90.97%) being the most abundant, followed by Proteus vulgaris (8.33%) and Proteus penneri (0.70%). The Proteus isolates displayed significant resistance to β-lactam antibiotics, with a Mean ± SD of 96.64 ± 22.73. A substantial portion of the Proteus spp. Isolated exhibited multidrug resistance (87.89%), with Proteus mirabilis (82.27%) being the most prevalent MDR species. Moreover, about 71.0% of the Proteus spp were ESBL producers, with Proteus mirabilis (64.54%) being the most predominant. Furthermore, 67.38% of all isolates exhibited MDR and ESBL production, and Proteus mirabilis (62.41%) was the most significant among the three Proteus species. These findings highlight the occurrence of multidrug resistance and ESBL production among Proteus spp. in Northeastern Nigeria, with Proteus mirabilis particularly noteworthy. This information is crucial for guiding clinical decision-making, especially in managing infections caused by multidrug-resistant and ESBL-producing Proteus strains.

Background Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae including Escherichia coli (E. coli), are recognized as a global public health threat due to their multidrug-resistant (MDR) phenotypes and their rapid dissemination in aquatic environments. Nevertheless, studies investigating the prevalence and antimicrobial resistance (AMR) profile of ESBL-producing E. coli in Lebanese surface water are limited. Objective This study aimed to assess the physicochemical properties and microbial contamination load and to determine the distribution of AMR patterns of ESBL-producing E. coli in surface water samples from different sites in the North Governorate of Lebanon. Methods Water samples were collected from 25 major sites in North Lebanon. These samples were analyzed for the presence of total coliforms, E. coli, and fecal enterococci. Phenotypic and genetic characterizations were then performed for E. coli isolates to determine their resistance patterns and phylogenetic groups. Results Fifty-six samples out of 100 samples were positive for ESBL-producing E. coli, mostly harboring blaCTX-M (40/56, 71%) including blaCTX-M-15 (33/40, 82%), blaTEM gene (36/56, 64%), blaSHV (20/56, 36%), and blaOXA (16/56, 29%) including blaOXA-48 gene (11/16, 69%). Most ESBL-producing E. coli isolates belonged to the extra-intestinal pathogenic phylogroup B2 (40/56, 71.4%) while 10/56 (17.9%) belonged to the commensal phylogroup A. Conclusion Our results highlight the need to implement effective water monitoring strategies to control transmission of ESBL-producing E. coli in surface water and thus reduce the burden on human and animal health.

Background and Aim: Bats are a reservoir for the pathogenic bacteria Klebsiella pneumoniae and can spread it through feces that fall in nests/caves, carried, and dropped while they fly near human settlements, and from their saliva. The emergence and spread of multidrug resistance (MDR) strains of K. pneumoniae indicate that resistant to antibiotics, especially extended-spectrum beta-lactamase (ESBL), is considered an important global health threat. The aim of this study was to determine the presence of the gene encoding extended beta-lactamase in K. pneumoniae isolated from fresh bat feces collected from the Tanjung Ringgit bat cave, East Lombok. Materials and Methods: In this study, 150 fresh fecal samples were analyzed using standard microbiological techniques for the presence of K. pneumoniae. K. pneumoniae-positive isolates were subjected to antibiotic sensitivity testing, followed by molecular detection using polymerase chain reaction. Results: This study showed that 14 (9.3%) of 150 samples were positive for K. pneumoniae. Ten of the 14 samples (71.4%) were MDR isolates and 6 (42.9%) had the blaSHV gene identified. Conclusion: The presence of K. pneumoniae isolated from fresh bat feces, which is MDR and has the blaSHV gene encoding ESBL indicates that bats can be a reservoir for the transmission of MDR and ESBL bacteria has an impact on public health in the study area.

Yanhua Liu,1,2,&ast; Fuxing Li,1,2,&ast; Youling Fang,2,3 Qiaoshi Zhong,1,2 Yanping Xiao,1,2 Yunwei Zheng,1,2 Junqi Zhu,2,3 Chuwen Zhao,2,3 Xingwei Cao,1,2 Jianqiu Xiong,4 Longhua Hu1,2 1The Second Affiliated Hospital, Jiangxi Medical College, Nanchang University, Jiangxi, Nanchang, People’s Republic of China; 2Department of Jiangxi Provincial Key Laboratory of Medicine, Clinical Laboratory of the Second Affiliated Hospital of Nanchang University, Jiangxi, Nanchang, People’s Republic of China; 3School of Public Health, Nanchang University, Jiangxi, Nanchang, People’s Republic of China; 4Department of Nursing, the Second Affiliated Hospital of Nanchang University, Jiangxi, Nanchang, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jianqiu Xiong, Department of Nursing, The Second Affiliated Hospital of Nanchang University, Mingde Road No. 1, Jiangxi, Nanchang, 330006, People’s Republic of China, Email 1773014059@qq.com Longhua Hu, Department of Jiangxi Provincial Key Laboratory of Medicine, Clinical Laboratory of the Second Affiliated Hospital of Nanchang University, Mingde Road No. 1, Jiangxi, Nanchang, 330006, People’s Republic of China, Email longhuahu@163.comObjective: This research aimed to analyze the clinical characteristics, prognosis, and antimicrobial treatment of bloodstream infections (BSI) caused by Enterobacter cloacae complex (ECC).Methods: The clinical data of patients with bloodstream infections caused by Enterobacter cloacae complex from April 2017 to June 2023 were collected retrospectively. These data were then analyzed in subgroups based on the detection results of extended-spectrum β-lactamase (ESBL), 30-day mortality, and the type of antimicrobial agent used (β-lactam/β-lactamase inhibitor combinations (BLICs) or carbapenems).Results: The proportion of ESBL-producing Enterobacter cloacae complex was 32.5% (37/114). Meanwhile, ICU admission, receiving surgical treatment within 3 months, and biliary tract infection were identified as risk factors for ESBL-producing ECC-BSI. Additionally, immunocompromised status and Sequential Organ Failure Assessment (SOFA) score ≥ 6.0 were identified as independent risk factors of 30-day mortality in patients with ECC-BSI (n = 108). Further analysis in BSI patients caused by non-ESBL-producing ECC revealed that patients treated with BLICs (n = 45) had lower SOFA scores and lower incidence of hypoproteinemia and sepsis compared with patients treated with carbapenems (n = 20). Moreover, in non-ESBL-producing ECC-BSI patients, the univariate Cox regression analysis indicated a significantly lower 30-day mortality rate in patients treated with BLICs compared to those treated with carbapenems (hazard ratios (HR) [95% CI] 0.190 [0.055– 0.662], P = 0.009; adjusted HR [95% CI] 0.106 [0.013– 0.863], P = 0.036).Conclusion: This study investigated the factors influencing the susceptibility to infection by ESBL-producing strains and risk factors for 30-day mortality in ECC-BSI patients. The results revealed that ESBL-negative ECC-BSI patients treated with BLICs exhibited significantly lower 30-day mortality compared to those treated with carbapenems. BLICs were found to be more effective in ECC-BSI patients with milder disease (ESBL-negative and SOFA ≤ 6.0).Keywords: Enterobacter cloacae complex, extended-spectrum β-lactamase, bloodstream infection, risk factors, β-lactam/β- lactamase inhibitor combinations, carbapenems

Antimicrobial stewardship focuses on identifying patients who require extended-spectrum beta-lactamase (ESBL)-targeted therapy. 'Rule-in' tools have been researched extensively in areas of low endemicity; however, such tools are inadequate for areas with high prevalence of ESBL-producing pathogens, as almost all patients will be selected. To develop a machine-learning-based 'rule-out' tool suitable for areas with high levels of resistance. Gradient-boosted decision trees were used to train and validate a risk prediction model on data from 17,913 (45% ESBL) patients with Escherichia coli and Klebsiella pneumoniae in urine cultures. The predictive power of different sets of variables was evaluated using Shapley values to evaluate the contributions of variables. The model successfully identified patients with low risk of ESBL resistance in ESBL-endemic areas (area under receiver operating characteristic curve 0.72). When used to select the 30% of patients with the lowest predicted risk, the model yielded a negative predictive value ≥0.74. A simplified model with seven input features was found to perform nearly as well as the full model. This simplified model is freely accessible as a web application. This study found that a risk calculator for antibiotic resistance can be a viable 'rule-out' strategy to reduce the use of ESBL-targeted therapy in ESBL-endemic areas. The robust performance of a version of the model with limited features makes the clinical use of such a tool feasible. This tool provides an important alternative in an era with growing rates of ESBL-producing pathogens, where some experts have called for empirical use of carbapenems as first-line therapy for all patients in areas with high prevalence of ESBL-producing pathogens. [ABSTRACT FROM AUTHOR]

Escherichia coli, a member of the commensal intestinal microbiota, is a significant aetiology of urinary tract infections (UTIs) and has a propensity for acquiring multidrug resistance characteristics, such as extended-spectrum beta-lactamases (ESBLs). Despite the increase in the incidence of ESBL-producing E. coli infections in sub-Saharan Africa, routine ESBL detection in Ghana is often absent, and molecular data on ESBL genotypes is scarce. Eleven ESBL-producing E. coli recovered from mid-stream urine samples were subjected to antimicrobial susceptibility testing and whole-genome sequence analyses. All isolates exhibited multidrug resistance, demonstrating phenotypic resistance to third-generation cephalosporins, such as cefotaxime, ceftazidime, and cefpodoxime. Three isolates demonstrated resistance to norfloxacin (a fluoroquinolone), and one isolate demonstrated intermediate resistance to ertapenem (a carbapenem). Analysis of the draft genomes identified multiple antimicrobial resistance genes including ESBL genotypes blaTEM-1B/TEM-190 (6/11 and 1/11, respectively), blaCTX-M-15/CTX-M-3 (7/11 and 1/11) and blaOXA-1/OXA-181 (3/11 and 1/11). The strains belong to 10 different serotypes and 10 different multilocus sequence types. This study provides information on phenotypic resistance in 11 ESBL E. coli from Ghana and AMR genotypes within their genomes. [ABSTRACT FROM AUTHOR]

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Objective: Urinary tract infections (UTIs) due to extended-spectrum betalactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae are among the leading causes of morbidity and mortality in older adults. Identifying associated factors for ESBL production may contribute to more appropriate empirical treatment. Materials and methods: This was a prospective observational study. Hospitalized patients of age > 65 with community-onset or hospital-acquired upper UTI due to E. coli or Klebsiella pneumoniae were included. A multivariate analysis was performed. Results: A total of 97 patients were included. ESBL prevalence among UTIs with E. coli or Klebsiella pneumoniae was 69.1% (n = 67). CRP values at the time of UTI diagnosis were found to be significantly higher in the ESBL-producing group (p = 0.004). The multivariate analysis revealed that male gender (OR: 2.72, CI: 1.02-7.25), prior recurrent UTI (OR: 3.14, CI: 1.21-8.14), and the development of secondary bacteremia (OR: 4.95, CI: 1.03-23.89) were major associated factors for UTI in older adults due to ESBL-producing E. coli and Klebsiella pneumoniae. Conclusion: Severe UTI in older men with a history of recurrent UTI may be a warning to the clinician for ESBL production in the setting of high ESBL prevalence. Carbapenems may be prioritized in the empirical treatment of patients with known risk factors for ESBL. [ABSTRACT FROM AUTHOR]

AbstractWe developed a prediction model for cefotaxime resistance in patients with K. pneumoniae bacteremia. Adult patients with K. pneumoniae bacteremia were grouped into derivation (from March 2018 to December 2019) and validation (from January 2020 to August 2020) cohorts. The prediction scoring system was based on factors associated with cefotaxime resistance identified by the logistic regression model. A total of 358 patients were enrolled (256 for derivation, 102 for validation). In the multivariable analysis, age ≥65 years, hospital-acquired infection, prior antimicrobial use, and an updated Charlson comorbidity index ≥3 points were associated with cefotaxime resistance in the derivation cohort. When each variable was counted as 1 point, the values of the area under the curve were 0.761 in the derivation and 0.781 in the validation cohorts. The best cutoff value using the Youden index was ≥2 with 73.6% sensitivity and 67.5% specificity. Our simple scoring system favorably predicted cefotaxime resistance. [ABSTRACT FROM AUTHOR]

ABSTRACT Klebsiella species commonly reside in dairy cattle guts and are consistently exposed to beta-lactam antibiotics, including ceftiofur, which are frequently used on the U.S. dairy farms. This may impose selection pressure and result in the emergence of extended-spectrum beta-lactamase (ESBL)-producing strains. However, information on the status and antimicrobial resistance (AMR) profile of ESBL-Klebsiella spp. in the U.S. dairy farms is largely unknown. This study aimed to determine the prevalence and AMR profile of ESBL-Klebsiella spp. and the factors affecting their occurrence in dairy cattle farms. Rectal fecal samples (n = 508) and manure, feed, and water samples (n = 64) were collected from 14 dairy farms in Tennessee. Samples were directly plated on CHROMagar ESBL, and presumptive Klebsiella spp. were confirmed using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibility testing was performed on the isolates against panels of 14 antimicrobial agents from 10 classes using minimum inhibitory concentration. Of 572 samples, 57 (10%) were positive for ESBL-Klebsiella spp. The fecal prevalence of ESBL-Klebsiella spp. was 7.2% (95% CI: 6.5–8.0). The herd-level fecal prevalence of ESBL-Klebsiella spp. was 35.7% (95% CI: 12.7–64.8). The fecal prevalence of ESBL-Klebsiella spp. was significantly higher in calves than in cows and higher in cows with higher parity (≥3) as compared to cows with low parity (P < 0.001). Most (96.5%, n = 57) ESBL-Klebsiella spp. were resistant to ceftriaxone. The highest level of acquired co-resistance to ceftriaxone in ESBL-Klebsiella spp. was to sulfisoxazole (66.7%; 38/57). About 19% of ESBL-Klebsiella spp. were multidrug resistant. The presence of ESBL-producing Klebsiella spp. in dairy cattle, feed, and water obtained from troughs could play a crucial epidemiological role in maintaining and spreading the bacteria on farms and serving as a point source of transmission.IMPORTANCEWe collected 572 samples from dairy farms, including rectal feces, manure, feed, and water. We isolated and identified extended-spectrum beta-lactamase (ESBL)-Klebsiella spp. and conducted an antimicrobial susceptibility test and analyzed different variables that may be associated with ESBL-Klebsiella spp. in dairy farms. The results of our study shed light on how ESBL-Klebsiella spp. are maintained through fecal-oral routes in dairy farms and possibly exit from the farm into the environment. We determine the prevalence of ESBL-Klebsiella spp. and their antimicrobial susceptibility profiles, underscoring their potential as a vehicle for multiple resistance gene dissemination within dairy farm settings. We also collected data on variables affecting their occurrence and spread in dairy farms. These findings have significant implications in determining sources of community-acquired ESBL-Enterobacteriaceae infections and designing appropriate control measures to prevent their spread from food animal production systems to humans, animals, and environments.

Background: The gut microbiome is made up of a diverse range of bacteria, especially gram-negative bacteria, and is crucial for human health and illness. There is a great deal of interest in the dynamic interactions between gram-negative bacteria and their host environment, especially considering antibiotic resistance. This work aims to isolate gram-negative bacteria that exist in the gut, identify their species, and use resistance-associated gene analysis to define their resistance mechanisms. Methods: Samples were collected from all patients who had a stool culture at a tertiary care center in Lebanon. Each type of bacteria that was identified from the stool samples was subjected to critical evaluations, and all discovered strains underwent antimicrobial susceptibility testing. Polymerase chain reaction was used to profile the genes for Carbapenem-resistant Enterobacteriaceae (CRE), Extended-spectrum beta-lactamase (ESBL), and that of Pseudomonas aeruginosa strains. Results: Escherichia coli, Klebsiella species, and Pseudomonas aeruginosa turned out to be the predominant microbiota members. Escherichia coli strains had a high frequency of extended-spectrum beta-lactamase genes, with the most discovered gene being bla CTX-M. Additionally, a considerable percentage of isolates had carbapenemase-resistant Enterobacteriaceae genes, suggesting the rise of multidrug-resistant strains. Multidrug resistance genes, such as bla mexR, bla mexB, and bla mexA, were found in strains of Pseudomonas aeruginosa, highlighting the possible difficulties in treating infections brought on by these bacteria. Conclusion: The findings highlight the critical importance of effective surveillance and response measures to maintain the effectiveness of antibiotics considering the introduction of multidrug resistance genes in Pseudomonas aeruginosa and ESBL and CRE genes in Escherichia coli.

ABSTRACT The aim of this study was to evaluate the proportion of resistance to a temocillin, tigecycline, ciprofloxacin, and chloramphenicol phenotype called t2c2 that resulted from mutations within the ramAR locus among extended-spectrum β-lactamases-Enterobacterales (ESBL-E) isolated in three intensive care units for 3 years in a French university hospital. Two parallel approaches were performed on all 443 ESBL-E included: (i) the minimal inhibitory concentrations of temocillin, tigecycline, ciprofloxacin, and chloramphenicol were determined and (ii) the genomes obtained from the Illumina sequencing platform were analyzed to determine multilocus sequence types, resistomes, and diversity of several tetR-associated genes including ramAR operon. Among the 443 ESBL-E strains included, isolates of Escherichia coli (n = 194), Klebsiella pneumoniae (n = 122), and Enterobacter cloacae complex (Ecc) (n = 127) were found. Thirty-one ESBL-E strains (7%), 16 K. pneumoniae (13.1%), and 15 Ecc (11.8%) presented the t2c2 phenotype in addition to their ESBL profile, whereas no E. coli presented these resistances. The t2c2 phenotype was invariably reversible by the addition of Phe-Arg-β-naphthylamide, indicating a role of resistance-nodulation-division pumps in these observations. Mutations associated with the t2c2 phenotype were restricted to RamR, the ramAR intergenic region (IR), and AcrR. Mutations in RamR consisted of C- or N-terminal deletions and amino acid substitutions inside its DNA-binding domain or within key sites of protein–substrate interactions. The ramAR IR showed nucleotide substitutions involved in the RamR DNA-binding domain. This diversity of sequences suggested that RamR and the ramAR IR represent major genetic events for bacterial antimicrobial resistance.IMPORTANCEMorbimortality caused by infectious diseases is very high among patients hospitalized in intensive care units (ICUs). A part of these outcomes can be explained by antibiotic resistance, which delays the appropriate therapy. The transferable antibiotic resistance gene is a well-known mechanism to explain the high rate of multidrug resistance (MDR) bacteria in ICUs. This study describes the prevalence of chromosomal mutations, which led to additional antibiotic resistance among MDR bacteria. More than 12% of Klebsiella pneumoniae and Enterobacter cloacae complex strains presented mutations within the ramAR locus associated with a dysregulation of an efflux pump called AcrAB-TolC and a porin: OmpF. These dysregulations led to an increase in antibiotic output notably tigecycline, ciprofloxacin, and chloramphenicol associated with a decrease of input for beta-lactam, especially temocillin. Mutations within transcriptional regulators such as ramAR locus played a major role in antibiotic resistance dissemination and need to be further explored.

Introduction: Extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase (AmpC-BL) in Enterobacteriaceae are a global threat. Coproduction of AmpC-BL and ESBL reduces therapeutic options, with more resistance against cephamycin and beta-lactamase inhibitor combinations. This study determines the proportion, incidence, and distribution of AmpC-BL-resistant genes in clinically isolated ESBL-producing Escherichia coli, completing its scarce data in Indonesia. Methods: The samples in this study were ESBL-producing E. coli from blood and urine specimens, confirmed by BD Phoenix semiautomatic examination combined with cefoxitin disk screening method. Confirmation tests of AmpC-BL used the AmpC-disk method and the polymerase chain reaction method. Results: Thirteen (27.8%) of 108 E. coli isolates were nonsusceptible to cefoxitin, and 3 (11.5%) isolates were confirmed to produce AmpC-BL and contained AmpC-BL CITM gene. Of the 3 AmpC-BL and ESBL coproduction isolates, one isolate was a copresentation of ESBL and AmpC-BL genes, namely blaTEM and CITM. Fisher’s exact test showed that the coproduction of AmpC-BL in ESBL-producing isolates was associated with reduced susceptibility to cefoxitin (P = 0.020) and amoxicillin-clavulanate (P = 0.048) compared to isolates producing ESBL alone. The effect of ESBL and AmpC-BL coproduction on reducing susceptibility to carbapenems needs further investigation. Conclusions: AmpC coproduction was found in 3 (2.8%) of 108 ESBL-producing E. coli isolates, and one isolate copresented AmpC-BL and ESBL coding genes blaTEM and CITM. These three isolates were associated with widened antibiotic resistance to cefoxitin and amoxicillin--clavulanate compared to isolates producing ESBL alone.

Abstract Extended-spectrum beta-lactamase (ESBL)-producing organisms are widely recognized as clinically relevant causes of difficult-to-treat infections. CTX-M has formed a rapidly growing family distributed worldwide among a wide range of clinical bacteria, particularly members of Enterobacteriaceae. Circulating banknotes, exchanged daily among people, pose a potential vehicle for transmitting multidrug resistance. We screened for ESBL-carrying bacteria in the present study and reported CTX-M mutations in Bangladesh's banknotes. We sequenced the genes and performed homology modeling using the Swiss model with CTX-M-15 (4HBT) as a template. Then, we performed molecular docking of mecillinam with the template and the generated model using Autodock 4.2 (Release 4.2.6). After docking, we visually inspected the complexes built using Autodock tools for polar contacts and pi-pi interactions in PyMOL 2.5.4. Our partially sequenced bla CTX-M was related to bla CTX-M-10 and bla CTX-M-15. We observed multiple single-nucleotide substitution mutations, i.e., G613T (silent mutation), A626T (I176F), and A503G (N135D). Homology modeling showed high similarity when the model was superimposed over the template. The orientation of Asn (135) in the template and Asp (135) in the model does not show a significant difference. Likewise, Ile (176) in the template and Phe (176) in the model offer the same orientation. Our generated model could bind to Lys237, Ser240, and Asp135 residues with the lowest binding energy on docking. Our predicted binding of the mecillinam to the mutated D-135 residue in the model indicates contributions and supports previous reports proposing CTX-M-15 to CTX-M-127 mutational conversion on the mecillinum resistance phenotype.

Background: In Gabon, studies on the characterization of extended-spectrum beta-lactamase-producing Escherichia coli in young children with diarrhoea are almost nonexistent. The objective was to evaluate the prevalence of antibiotic resistance to extended-spectrum beta-lactamase-producing Escherichia coli in children at public hospitals in Franceville, Gabon. Methods: Seventy diarrhoea faecal samples were collected from children aged 0–5 years. The culture and isolation of colonies were carried out on MacConkey agar. The colonies were identified using VITEK 2. The determination of the extended-spectrum beta-lactamase’s profiles was accomplished using the double disk method. The identification of phylogroups and pathotypes was performed by PCR. Identification of the ESBL genes was performed by sequencing. Results: A total of 26 strains of Escherichia coli (33.0%) were identified from 78 bacterial isolates. Twenty (77.0%) Escherichia coli strains carried extended-spectrum beta-lactamases blaCTX-M-15 and 5.0% carried blaSHV-12 subtypes. Phylogroup D (62.0%) was predominant, followed by B1 (12.0%), B2 (8.0%) and E (4.0%). The bacterial pathogens causing diarrhoea were enterohemorrhagic E. coli (12.0%), typical enteropathogenic Escherichia coli (8.0%), atypical enteropathogenic Escherichia coli (4.0%), Enteroaggregative Escherichia coli (4.0%) and enteroinvasive E. coli (4.0%). Conclusions: This study showed a high prevalence of extended-spectrum beta-lactamase, Escherichia coli of phylogroup D and pathotype enterohemorrhagic Escherichia coli in children under 5 years old in public hospitals in Franceville, most probably due to the misuse or inappropriate consumption of beta-lactams.

Background/Objectives: Over the past few decades, extended-spectrum β-lactamase (ESBL)-producing bacteria have become a great concern in healthcare systems worldwide, imposing large burdens by increasing antimicrobial resistance and patient morbidity. Given the high mortality rates and emergence of multidrug-resistant (MDR) strains, monitoring ESBL prevalence and resistance patterns is crucial. This study aimed to evaluate ESBL-producing Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae over three years, focusing on phenotypic distribution and resistance profiles. Methods: A total of 1599 ESBL-producing bacterial samples were collected and analysed. A panel of 20 antibiotics was tested to determine resistance traits. Data were recorded on phenotypical distribution, isolation types, changes in antibiotic resistance, and the relation of such changes to antibiotic consumption (defined daily dose) from clinical isolates. Results: Phenotypical analysis revealed the minimal presence of the Cefotaximase from Munich (CTX-M) phenotype in E. coli and K. pneumoniae, creating a distinct epidemiological profile compared to global patterns. Shifts in isolation trends, particularly in P. mirabilis, suggest an expected increase in associated-mortality-rate in the coming years. While resistance trends were not statistically significant, MDR and extensively drug-resistant (XDR) strains were identified across all three bacteria. Only meropenem showed consistent 100% efficacy against E. coli, with other antibiotics displaying only partial effectiveness. Conclusions: These findings highlight the need for ongoing surveillance of ESBL-producing bacteria and underscore challenges in managing antibiotic resistance due to limited efficacy of last-resort treatments. The unique phenotypical distribution observed could impact local resistance management strategies in hospital settings in the coming years.

Background: Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae including Escherichia coli (E. coli), are recognized as a global public health threat due to their multidrug-resistant (MDR) phenotypes and their rapid dissemination in aquatic environments. Nevertheless, studies investigating the prevalence and antimicrobial resistance (AMR) profile of ESBL-producing E. coli in Lebanese surface water are limited. Objective: This study aimed to assess the physicochemical properties and microbial contamination load and to determine the distribution of AMR patterns of ESBL-producing E. coli in surface water samples from different sites in the North Governorate of Lebanon. Methods: Water samples were collected from 25 major sites in North Lebanon. These samples were analyzed for the presence of total coliforms, E. coli , and fecal enterococci. Phenotypic and genetic characterizations were then performed for E. coli isolates to determine their resistance patterns and phylogenetic groups. Results: Fifty-six samples out of 100 samples were positive for ESBL-producing E. coli , mostly harboring blaCTX-M (40/56, 71%) including blaCTX-M-15 (33/40, 82%), blaTEM gene (36/56, 64%), blaSHV (20/56, 36%), and blaOXA (16/56, 29%) including blaOXA-48 gene (11/16, 69%). Most ESBL-producing E. coli isolates belonged to the extra-intestinal pathogenic phylogroup B2 (40/56, 71.4%) while 10/56 (17.9%) belonged to the commensal phylogroup A. Conclusion: Our results highlight the need to implement effective water monitoring strategies to control transmission of ESBL-producing E. coli in surface water and thus reduce the burden on human and animal health. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is a global health issue that affects humans, animals, and agriculture. This study assessed the prevalence and antimicrobial susceptibility of extended-spectrum betalactamase-producing Escherichia coli (ESBL-EC) in poultry and hand swabs of meat handlers working in a dressing plant in the Philippines. Fourteen (14) hand swabs from poultry handlers and 89 poultry cecal samples were collected and screened for ESBL production using MacConkey agar supplemented with cefotaxime and eosin methylene blue agar. Identification of E. coli through biochemical testing and antimicrobial susceptibility testing was then performed using the VITEK 2 System. Results showed that 7.14% (1/14) of the hand swabs and 23.60% (21/89) of the poultry cecal samples were positive for ESBL-EC. Identified isolates were further evaluated for their antimicrobial susceptibility using 33 antibiotics from various groups. Results showed 100% (22/22) resistance to the following antimicrobial drugs: ampicillin, ticarcillin, cefalotin, cefuroxime, cefuroxime axetil, cefixime, cefotaxime, nalidixic acid, ciprofloxacin, levofloxacin, moxifloxacin, ofloxacin, tetracycline, trimethoprim, and trimethoprim/sulfamethoxazole. Resistance of select ESBL-EC isolates against other drugs under the beta-lactam, tetracycline, chloramphenicol, and aminoglycoside groups were also observed. Antimicrobial-resistant bacteria may be found in both human and animal sources, which highlights the need for ensuring safety practices to prevent their spread in the community. Gene sequencing and phylogenetic analysis may be used to further evaluate this relationship. Continuous surveillance and review of standards and protocols are also needed to monitor antimicrobial resistance, as underlined by the strategies of the One Health approach. [ABSTRACT FROM AUTHOR]

Background and Objectives: Klebsiella pneumoniae is an opportunistic pathogen responsible for causing nosocomial and community-acquired infections. Its pathogenicity is associated with a variety of virulence factors and antibiotic resistance. The aim of the present study was to compare virulence attributes between ESBL and non-ESBL producing isolates. Materials and Methods: A total of 113 K. pneumoniae including 56 ESBL and 57 non ESBL-producers were collected in Bushehr province, Iran, from November 2017 to February 2019. Enzymatic profile, hypermucoviscosity and biofilm formation were investigated phenotypically. In addition, the presence of rmpA, aerobactin, kfu, allS, mrkD, ybtS, entB, iutA, fimH, wabG, wcaG, K1 and K2 genes were detected by PCR and sequencing. Results: There was no statistically significant difference in enzymatic profile between ESBL and non-ESBL producers. The prevalence of the hypermocoviscosity was lower among ESBL compared to non-ESBL producers but the intensity of biofilm was higher in the ESBL producers. Among the virulence genes, K1, rmpA, iutA, and aero were observed only in non-ESBLs. Moreover, the carriage of allS, K, K2, rmpA, iutA and aero genes was higher in hypermucoviscous in comparison with non hypermucoviscous isolates. Conclusion: The identification of potentially pathogenic isolates plays an important role in preventing their spread as well as the success of their treatment. [ABSTRACT FROM AUTHOR]

Despite the significant impact of multi-drug-resistant bacteraemia, especially extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) and carbapenem-resistant Enterobacterales (CRE), the burden of disease has not been investigated thoroughly. To evaluate the clinical outcomes and socio-economic burden of ESBL-E and CRE bacteraemia nationwide in the Republic of Korea. A search was undertaken for all cases of ESBL-E and CRE bacteraemia and matched controls in 10 hospitals in the Republic of Korea over 6 months. Patients with ESBL-E or CRE bacteraemia were classified as the R group, and matched controls with antibiotic-susceptible bacteraemia and without infection were classified as the S and N groups, respectively. Patients' clinical data were collected, and the economic burden was estimated based on medical expenses, loss of productivity and total costs. In total, 795 patients were identified, including 265 patients with ESBL-E or CRE bacteraemia and their matched controls. The mean total length of stay for patients with ESBL-E and CRE in the R group was 1.53 and 1.90 times that of patients in the S group, respectively. The 90-day mortality rates for ESBL-E in the R and S groups were 12.1% and 5.6%, respectively, and the corresponding figures for CRE were 28.6% and 12.0%. There were significant differences in the total costs between the R, S and N groups for both ESBL-E and CRE (ESBL-E: $11,151 vs $8712 vs $6063, P =0.004; CRE: $40,464 vs $8748 vs $7279, P =0.024). The clinical and economic burden imposed by ESBL-E or CRE bacteraemia was extremely high. These findings suggest that efforts to control resistant bacteraemia are necessary to reduce this burden. [ABSTRACT FROM AUTHOR]

Among carbapenem-resistant Enterobacterales (CRE) are diverse mechanisms, including those that are resistant to meropenem but susceptible to ertapenem, adding further complexity to the clinical landscape. This study investigates the emergence of ertapenem-resistant, meropenem-susceptible (ErMs) Escherichia coli and Klebsiella pneumoniae CRE across five hospitals in San Antonio, Texas, USA, from 2012 to 2018. The majority of the CRE isolates were non-carbapenemase producers (NCP; 54%; 41/76); 56% of all NCP isolates had an ErMs phenotype. Among ErMs strains, E. coli comprised the majority (72%). ErMs strains carrying blaCTX-M had, on average, 9-fold higher copies of blaCTX-M than CP-ErMs strains as well as approximately 4-fold more copies than blaCTX-M-positive but ertapenem- and meropenem-susceptible (EsMs) strains (3.7 vs. 0.9, p < 0.001). Notably, carbapenem hydrolysis was observed to be mediated by strains harboring blaCTX-M with and without a carbapenemase(s). ErMs also carried more mobile genetic elements, particularly IS26 composite transposons, than EsMs (37 vs. 0.2, p < 0.0001). MGE- ISVsa5 was uniquely more abundant in ErMs than either EsMs or ErMr strains, with over 30 more average ISVsa5 counts than both phenotype groups (p < 0.0001). Immunoblot analysis demonstrated the absence of OmpC expression in NCP-ErMs E. coli, with 92% of strains lacking full contig coverage of ompC. Overall, our findings characterize both collaborative and independent efforts between blaCTX-M and OmpC in ErMs strains, indicating the need to reappraise the term "non-carbapenemase (NCP)", particularly for strains highly expressing blaCTX-M. To improve outcomes for CRE-infected patients, future efforts should focus on mechanisms underlying the emerging ErMs subphenotype of CRE strains to develop technologies for its rapid detection and provide targeted therapeutic strategies. [ABSTRACT FROM AUTHOR]

ObjectiveUrinary tract infections (UTIs) due to extended-spectrum beta-lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae are among the leading causes of morbidity and mortality in older adults. Identifying associated factors for ESBL production may contribute to more appropriate empirical treatment.Materials and methodsThis was a prospective observational study. Hospitalized patients of age > 65 with community-onset or hospital-acquired upper UTI due to E. coli or Klebsiella pneumoniae were included. A multivariate analysis was performed.ResultsA total of 97 patients were included. ESBL prevalence among UTIs with E. coli or Klebsiella pneumoniae was 69.1% (n = 67). CRP values at the time of UTI diagnosis were found to be significantly higher in the ESBL-producing group (p = 0.004). The multivariate analysis revealed that male gender (OR: 2.72, CI: 1.02–7.25), prior recurrent UTI (OR: 3.14, CI: 1.21–8.14), and the development of secondary bacteremia (OR: 4.95, CI: 1.03–23.89) were major associated factors for UTI in older adults due to ESBL-producing E. coli and Klebsiella pneumoniae.ConclusionSevere UTI in older men with a history of recurrent UTI may be a warning to the clinician for ESBL production in the setting of high ESBL prevalence. Carbapenems may be prioritized in the empirical treatment of patients with known risk factors for ESBL.

Background and Aim: Antimicrobial resistance (AMR) is becoming a public health concern. Foodborne pathogens are infectious agents that can be transmitted from animals to humans through food and can become resistant due to misuse and overuse of antibiotics, especially in poultry. This study aimed to detect the prevalence of multidrug-resistant and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from local and broiler chickens at the Cibinong market, West Java, Indonesia. Materials and Methods: A total of 60 cloacal swab samples from 30 local and broiler chickens sold at the Cibinong market in West Java were obtained by random sampling. From these samples, 39 E. coli isolates were obtained after being cultured on eosin methylene blue agar and molecularly identified using polymerase chain reaction (PCR). Six antibiotic disks were used for the antibiotic sensitivity test against E. coli isolates cultured on Mueller-Hinton agar. PCR was performed to detect ESBL genes (blaTEM, blaSHV, and blaCTX-M). Results: A total of 76.47% (39/51) cloacal swab samples were positive for E. coli. All E. coli isolates were sensitive to imipenem (100%), and 38 isolates were sensitive to cefoxitin (FOX) (97.4%). On average, the isolates were sensitive to amoxicillin-clavulanic acid (AMC) (69.2%) and ceftriaxone (CRO) (89.7%). E. coli isolates were occasionally resistant to enrofloxacin (25.64%), followed by gentamicin (20.51%), CRO (10.25%), AMC (7.69%), and FOX (2.56%). The prevalence of E. coli AMR was 10.25% (4/39). All four multidrug-resistant E. coli isolates (blaTEM and blaCTX-M) were confirmed to have the ESBL gene based on PCR. Conclusion: The prevalence of multidrug-resistant and ESBL-producing E. coli is still found, proving that there is still inappropriate use of antibiotics and a need for strict supervision of their use, especially around Cibinong market, West Java.

Background and Aim: Houseflies (Musca domestica) are synanthropic insects that are vectors of a wide range of multidrug-resistant pathogens responsible for infectious diseases. The aim of this study was to detect and characterize multidrug-resistant Enterobacteriaceae carried by houseflies in Bobo-Dioulasso. Materials and Methods: A total of 500 houseflies were captured in hospital and non-hospital environments in the city of Bobo-Dioulasso. For bacteriological analysis, they were divided into 125 batches of five flies each. Multidrug-resistant bacteria isolated on MacConkey agar supplemented with 4 μg/mL cefotaxime were identified on the basis of biochemical characteristics. Antibiotic susceptibility profiles were determined using the agar diffusion method. blaCTX-M resistance genes and quinolone resistance genes (plasmid-mediated quinolone resistance) were detected by conventional polymerase chain reaction. Results: Among 115 bacterial strains obtained, 26 were extended-spectrum beta-lactamase (ESBL)-producing enterobacteria: Escherichia coli (15), Klebsiella pneumoniae (6), Enterobacter cloacae (4), and Morganella morganii (1). Carriers were statistically more important in hospitals (12/26, p = 0.03). No carbapenem-resistance strains were observed. We identified ESBL resistance genes (Cefotaximase Munich; CTX-M group 1) (25/26) and quinolone resistance genes (QnrS) (6/26). Conclusion: Houseflies in the city of Bobo-Dioulasso are vectors for the transmission of multidrug-resistant enterobacteria. There is a need to monitor the associated risks for public health.

Background: Over the past decades, antibiotic-resistant Gram-negative bacteria commonly Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae have increased significantly. These microorganisms have great clinical importance because they increase hospital stay of the patients in the intensive care unit (ICU) leading to high morbidity and mortality. Because of their role in increasing morbidity and mortality, this study was performed to isolate extended-spectrum beta-lactamase (ESBL) producing Gram-negative bacilli screened by phenotypical method and further projected into molecular characterization by polymerase chain reaction. Aims and Objectives: The aims and objectives are to isolate the Gram-negative multidrug-resistant strains from clinically suspected bacterial infections in patients of neonatal, sick newborn, and pediatric ICU and to study antibiotic sensitivity pattern of isolated Gram-negative multidrug-resistant strains with special reference to molecular characterization. Materials and Methods: A total of 100 Gram-negative bacilli were isolated. Screening of ESBL positivity was done by double-disk synergy test (combined disc test method). Their antibiogram profile was interpreted. With the use of designed primers, 26 ESBL isolates each of E. coli and Klebsiella spp. were processed for molecular analysis of beta-lactamase family genes TEM and CTX-M. Results: Within the 100 samples, majority of the isolates (45%) were Klebsiella spp. and 40% was E. coli isolates. Highest ESBL-producing organisms were observed within E. coli (65%). Prevalence bla-TEM gene was highest followed by bla-CTX-M. These ESBL-producing organisms were found to be resistant to multiple classes of antibiotics. With extensive ESBL surveillance and proper usage of antibiotics, this threatening rise of antibiotic resistance can be mitigated. Conclusion: Gram-negative isolates showed high resistance to commonly used antibiotics. Significant proportions of them were MDR strains. Such high antibiotic resistance is associated with significant morbidity and mortality among pediatric population. MDR along with possession of ESBL associated resistance genes among Gram-negative bacilli pose a serious problem in therapeutic management of patients. Our study signifies that there is a high probability of Gram- negative bacilli to be multi-drug resistant and ESBL positive and earliest detection of such cases should be made.

ABSTRACT: Objectives: The production of expanded-spectrum beta-lactamases (ESBLs) and fluoroquinolone resistance in Enterobacteriaceae has become a global concern. The aim of this study was to investigate the spread of ESBL-producing and fluoroquinolone-resistant avian pathogenic Escherichia coli (APEC) in Kagoshima, a prefecture with the largest amount of poultry in Japan. Methods: The antimicrobial susceptibility and genetic characteristics of 228 APEC strains isolated from 57 farms in Kagoshima Prefecture, Japan, between 2005 and 2017 were analysed. Information about the companies with hatcheries connected to the farms was also collected, and the epidemiologic relatedness of APEC strains and the processes of adopting chicks were compared. Results: Seven CTX-M-type ESBL genes, blaCTX-M-1, blaCTX-M-2, blaCTX-M-14, blaCTX-M-15, blaCTX-M-25, blaCTX-M-55, and blaCTX-M-65, were found in 60 (26.3%) of the 228 APEC strains. The ciprofloxacin-resistant strains belonged to 10 different sequence types (ST10, ST23, ST93, ST155, ST156, ST350, ST359, ST602, ST648, and ST9479), and the two ST602 strains showed remarkably high ciprofloxacin resistance (MIC: 128 µg/mL) and had amino acid mutations in GyrA (S83L and D87N), ParC (S80I), and ParE (E460A). A CTX-M-55-type ESBL-producing fluoroquinolone-resistant Og78-ST23 strain was isolated multiple times over two years on a farm. Furthermore, epidemiologically closely related strains were isolated from different farms that used the same common hatcheries. Conclusions: APEC is often transferred from hatcheries to farms via healthy chicks, and the prudent use of antimicrobials and careful monitoring of resistant strains on poultry farms and hatcheries are important in preventing the selection and spread of high-risk APEC strains such as CTX-M-55-type ESBL-producing Og78-ST23.

Background and Aim: Antibiotic resistance in Klebsiella pneumoniae, especially extended-spectrum beta-lactamase (ESBL) producers, has become a global public health problem. This study aimed to estimate the prevalence of multidrug resistance (MDR) and ESBL-producing K. pneumoniae in wastewater from dairy farms. Materials and Methods: This study was conducted on dairy farms in East Java Province from June to October 2022. In total, 342 dairy farm wastewater samples were isolated on buffered peptone water media from six cities/regencies with the highest dairy cattle population in East Java. Samples were identified using MacConkey agar media, Gram-staining,eosin-methylene blue agar, and biochemical tests. In total, 14.32% (49/342) samples contained K. pneumoniae. Positive isolates were tested for antibiotic sensitivity. Klebsiella pneumoniae resistant to beta-lactam was confirmed using the double-disk synergy test to confirm the presence of ESBL-producing bacteria. Results: The percentage of antibiotic resistance in K. pneumoniae was 98% resistance to ampicillin, 67.3% to cefotaxime, 46.9% to tetracycline, 49% to ciprofloxacin, 98% to streptomycin, 14.3% to sulfamethoxazole–trimethoprim, and 83.7% to chloramphenicol. The prevalence of MDR in K. pneumoniae was 12.57% (43/342), with the highest prevalence in the five classes of antibiotics at 41.86% (18/43), and the prevalence of ESBL-producing K. pneumoniae was 5.55% (19/342), with the highest prevalence in the districts of Blitar and Pasuruan at 26.31% (5/19). Conclusion: Although the prevalence of ESBL-producing K. pneumoniae in wastewater samples from dairy farms was low, caution is recommended because they can be a reservoir for ESBL.

Wubalech Temsegen,1,2 Kasahun Gorems,2,3 Mekidim Mekonnen,1 Diriba Fufa,4 Tesfaye Kassa1 1School of Medical Laboratory Science, Faculty of Health Sciences, Jimma University, Jimma, Ethiopia; 2Microbiology Diagnostic Laboratory Unit of Jimma Medical Center, Jimma, Ethiopia; 3Department of Microbiology, Immunology and Parsitology, St Paul’s Hospital Millennium Medical College, Addis Ababa, Ethiopia; 4Department of Pediatrics & Child Health, School of Medicine, Jimma University, Jimma, EthiopiaCorrespondence: Kasahun Gorems, Department of Microbiology, Immunology and Parsitology, St Paul’s Hospital Millennium Medical College, P. O. Box, 1271, Addis Ababa, Ethiopia, Tel +251925494531, Email kasahungorems63@gmail.com; kasahun.gorems@sphmmc.edu.et Tesfaye Kassa, School of Medical Laboratory Sciences, Jimma University, P. O. Box 788, Jimma, Ethiopia, Tel +251931057195, Email ktes36@gmail.com; tesfaye.kassa@ju.edu.etPurpose: Extended-spectrum β-lactamase (ESBL) and carbapenemase-producing Enterobacteriaceae (CPE) are among the major threats to global health because of their encoded protection against key antibiotics.Methods: A comparative cross-sectional study was conducted among oncology and non-oncology patient groups (1:1; n = 214) on a consecutive sampling approach. Stool or rectal swab was collected from June 2021 to November 2021 and screened for ESBL-PE and CPE using ChromID-ESBL media. Confirmation for the enzymes was made by using combination disc and modified carbapenem inactivation methods, respectively. Disk diffusion method was used to determine antimicrobial susceptibility testing following the recommendations of CLSI 2022. SPSS software version 23 was used for data analysis.Results: Fecal carriage prevalence of ESBL-PE was found in 90 (84.1%) of oncology participants and in 77 (71.9%) of non-oncology patients (p = 0.032). Escherichia coli was the most common ESBL-PE isolate in 82 (62.5%) and 68 (88.3%) of oncology and non-oncology patients, followed by Klebsiella oxytoca [15 (11.5%) versus 6 (7.8%)], respectively. Out of the total ESBL-PE isolates from both oncology and non-oncology patient groups, the maximum level of resistance was observed against ciprofloxacin 177 (86.3%), trimethoprim-sulfamethoxazole 103 (80.3%), tetracycline 97 (75.8%), whereas enhanced susceptibility was appreciated to tigecycline 200 (97.6%), meropenem 162 (79.0%), and ertapenem 145 (70.7%) with no significant difference between oncology and non-oncology group. Carbapenemase-producing isolates from oncology patients were 12 (11.2%), whereas it was 4 (3.7%) (p = 0.611) from non-oncology group. Bacterial isolates from oncology in this study showed a trend of multiple drug resistance of 113 (88.3%).Conclusion: The results revealed alarmingly high carriage rates of ESBL and CPE among all study participants. Moreover, the isolates showed increased resistance rates to alternative drugs and had multiple antibiotic-resistant patterns. Hence, it is important to emphasize strict adherence to antimicrobial stewardship program as well as infection prevention and control practices.Keywords: carriage rate, oncology, extended-spectrum beta-lactamase, carbapenemase, Enterobacteriaceae, Ethiopia

[This corrects the article DOI: 10.3389/fmicb.2021.709418.].

BackgroundThe rapid spread of extended-spectrum beta-lactamase-producing E. coli (ESBL-EC) is an urgent global health threat. We examined child caretaker knowledge, attitudes, and practices (KAP) towards proper antimicrobial agent use and whether certain KAP were associated with ESBL-EC colonization of their children.MethodsChild caretakers living in semi-rural neighborhoods in peri-urban Quito, Ecuador were visited and surveyed about their KAP towards antibiotics. Fecal samples from one child (less than 5 years of age) per household were collected at two time points between July 2018 and May 2019 and screened for ESBL-EC. A repeated measures analysis with logistic regression was used to assess the relationship between KAP levels and child colonization with ESBL-EC.ResultsWe analyzed 740 stool samples from 444 children living in households representing a range of environmental conditions. Of 374 children who provided fecal samples at the first household visit, 44 children were colonized with ESBL-EC (11.8%) and 161 were colonized with multidrug-resistant E. coli (43%). The prevalences of ESBL-EC and multidrug-resistant E. coli were similar at the second visit (11.2% and 41.3%, respectively; N = 366). Only 8% of caretakers knew that antibiotics killed bacteria but not viruses, and over a third reported that they "always" give their children antibiotics when the child's throat hurts (35%). Few associations were observed between KAP variables and ESBL-EC carriage among children. The odds of ESBL-EC carriage were 2.17 times greater (95% CI: 1.18-3.99) among children whose caregivers incorrectly stated that antibiotics do not kill bacteria compared to children whose caregivers correctly stated that antibiotics kill bacteria. Children from households where the caretaker answered the question "When your child's throat hurts, do you give them antibiotics?" with "sometimes" had lower odds of ESBL-EC carriage than those with a caretaker response of "never" (OR 0.48, 95% CI 0.27-0.87).ConclusionCaregivers in our study population generally demonstrated low knowledge regarding appropriate use of antibiotics. Our findings suggest that misinformation about the types of infections (i.e. bacterial or viral) antibiotics should be used for may be associated with elevated odds of carriage of ESBL-EC. Understanding that using antibiotics is appropriate to treat infections some of the time may reduce the odds of ESBL-EC carriage. Overall, however, KAP measures of appropriate use of antibiotics were not strongly associated with ESBL-EC carriage. Other individual- and community-level environmental factors may overshadow the effect of KAP on ESBL-EC colonization. Intervention studies are needed to assess the true effect of improving KAP on laboratory-confirmed carriage of antimicrobial resistant bacteria, and should consider community-level studies for more effective management.

BackgroundThe prevalence of infections caused by extended-spectrum beta-lactamase producing Escherichia coli (ESBL-E. coli) is increasing worldwide, but the setting in which this increase is occurring is not well defined. We compared trends and risk factors for ESBL-E. coli bacteriuria in community vs healthcare settings.MethodsWe collected electronic health record data on all patients with E. coli isolated from urine cultures in a safety-net public healthcare system from January 2014 to March 2020. All analyses were stratified by healthcare-onset/associated (bacteriuria diagnosed > 48 h after hospital admission or in an individual hospitalized in the past 90 days or in a skilled nursing facility resident, N = 1277) or community-onset bacteriuria (bacteriuria diagnosed  65 (RR 1.88, CI 1.17, 3.05), male gender (RR 2.12, CI 1.65, 2.73), and Latinx race/ethnicity (RR 1.52, CI 0.99, 2.33) were associated with community-onset ESBL-E. coli. Only male gender (RR 1.53, CI 1.03, 2.26) was associated with healthcare-onset/associated ESBL-E. coli.ConclusionsESBL-E. coli bacteriuria frequency increased at a faster rate in healthcare-associated settings than in the community between 2014 and 2020. Male gender was associated with ESBL-E. coli bacteriuria in both settings, but additional risks-age > 65 and Latinx race/ethnicity-were observed only in the community.

Background and Aim: Antimicrobial-resistant microorganisms (ARMs) have been increasing among wild animals. Interactions occurring at the interface between wildlife, humans, and livestock can lead to the transmission of ARMs. Thus, the prevalence of ARMs in wild and domestic animals should be determined to address and prevent this issue. This study aimed to determine the resistance patterns of cefotaxime (CTX)-resistant Escherichia coli and identify the presence of extended-spectrum beta-lactamase (ESBL) genes in ESBL-producing E. coli among a population of wild banteng (Bos javanicus) and domestic cattle kept on farms located close to the Lam Pao non-hunting area, Kalasin province, Thailand. Materials and Methods: Forty-five fecal samples were taken from wild bantengs inhabiting the Lam Pao non-hunting area in Thailand, alongside 15 samples from domestic cattle. Bacterial culture, triple sugar iron, and motile indole lysine tests were conducted to identify E. coli. A polymerase chain reaction (PCR) was conducted for specific confirmation. MacConkey agar supplemented with 2 μg/mL of CTX was used to identify CTX-resistant E. coli, which would be used to identify ESBL production based on a double-disk synergy test. Extended-spectrum beta-lactamase-producing samples were subjected to disk diffusion tests to determine resistant patterns, and the sizes of PCR bands and DNA sequencing were used to differentiate ESBL gene types. Results: All samples tested positive for E. coli. Forty-five isolates from 15 banteng samples and three isolates from one domestic cattle sample displayed CTX-resistant and ESBL-producing traits. The banteng and domestic cattle populations exhibited nine and three distinct resistant patterns, respectively. The PCR results indicated that the banteng isolates harbored the following genes: Cefotaxime-M1 (n = 38), CTX-M9 (n = 5), and the SHV group (n = 2). All three isolates from the domestic cattle sample contained the CTX-M1 gene. Classification of ESBL genes based on the DNA sequences of the banteng isolates showed the characteristics of CTX-M15 (n = 20), CTX-M55 (n = 6), CTX-M14 (n = 5), and CTX-M79 (n = 1). The three domestic cattle isolates exhibited the characteristics of CTX-M15, CTX-M55, and CTX-M79. Conclusion: Despite no previous antibiotic applications, approximately one-third of the banteng samples displayed CTX resistance, indicating ARM contamination within the ecosystem. The similarity in ESBL genes between the banteng and domestic cattle populations suggests potential gene transmissions between these animal groups. However, the initial source of ARMs remains unclear, as the banteng population exhibited more ESBL genes than the domestic cattle, suggesting the possibility of multiple ARM sources. These findings raise concerns because the banteng population inhabits an area that is an important source of freshwater and nourishes the entire north-east region of Thailand and other South-east Asian countries, including Laos, Cambodia, and Southern Vietnam.

Background and Objectives: Klebsiella pneumoniae is an opportunistic pathogen responsible for causing nosocomial and community-acquired infections. Its pathogenicity is associated with a variety of virulence factors and antibiotic resistance. The aim of the present study was to compare virulence attributes between ESBL and non-ESBL producing isolates. Materials and Methods: A total of 113 K. pneumoniae including 56 ESBL and 57 non ESBL-producers were collected in Bushehr province, Iran, from November 2017 to February 2019. Enzymatic profile, hypermucoviscosity and biofilm formation were investigated phenotypically. In addition, the presence of rmpA, aerobactin, kfu, allS, mrkD, ybtS, entB, iutA, fimH, wabG, wcaG, K1 and K2 genes were detected by PCR and sequencing. Results: There was no statistically significant difference in enzymatic profile between ESBL and non-ESBL producers. The prevalence of the hypermocoviscosity was lower among ESBL compared to non-ESBL producers but the intensity of biofilm was higher in the ESBL producers. Among the virulence genes, K1, rmpA, iutA, and aero were observed only in non-ESBLs. Moreover, the carriage of allS, K, K2, rmpA, iutA and aero genes was higher in hypermucoviscous in comparison with non hypermucoviscous isolates. Conclusion: The identification of potentially pathogenic isolates plays an important role in preventing their spread as well as the success of their treatment.

Introduction: Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae are on the WHO priority pathogens list because they are associated with high mortality, health-care burden, and antimicrobial resistance (AMR), a serious problem that threatens global public health and should be addressed through the One Health approach. Non-human primates (NHP) have a high risk of acquiring these antibiotic-resistant bacteria due to their close phylogenetic relationship with humans and increased anthropogenic activities in their natural environments. This study aimed to detect and analyze the genomes of ESBL-producing Escherichia coli (ESBL-producing E. coli) in NHP from the Peruvian Amazon. Materials and methods: We collected a total of 119 fecal samples from semicaptive Saguinus labiatus, Saguinus mystax, and Saimiri boliviensis, and captive Ateles chamek, Cebus unicolor, Lagothrix lagothricha, and Sapajus apella in the Loreto and Ucayali regions, respectively. Subsequently, we isolated and identified E. coli strains by microbiological methods, detected ESBL-producing E. coli through antimicrobial susceptibility tests following CLSI guidelines, and analyzed their genomes using previously described genomic methods. Results: We detected that 7.07% (7/99) of E. coli strains: 5.45% (3/55) from Loreto and 9.09% (4/44) from Ucayali, expressed ESBL phenotype. Genomic analysis revealed the presence of high-risk pandemic clones, such as ST10 and ST117, carrying a broad resistome to relevant antibiotics, including three blaCTX-M variants: blaCTX-M-15, blaCTX-M-55, and blaCTX-M-65. Phylogenomic analysis confirmed the clonal relatedness of high-risk lineages circulating at the human-NHP interface. Additionally, two ESBL-producing E. coli strains were identified as EPEC (eae) and ExPEC according to their virulence profiles, and one more presented a hypermucoviscous phenotype. Discussion: We report the detection and genomic analysis of seven ESBLproducing E. coli strains carrying broad resistome and virulence factors in NHP from two regions of the Peruvian Amazon. Some of these strains are closely related to high-risk pandemic lineages previously reported in humans and domestic animals, highlighting the negative impact of anthropogenic activities on Amazonian wildlife. To our knowledge, this is the first documentation of ESBL-producing E. coli in NHP from the Amazon, underscoring the importance of adopting the One Health approach to AMR surveillance and minimizing the potential transmission risk of antibiotic-resistant bacteria at the human-NHP interface. [ABSTRACT FROM AUTHOR]