ESBL Producers Research Articles

Phytochemical Screening and Antimicrobial Activity of Methanol Extracts of Usteria guineensis and Sphaerocoryne gracilipes on Pathogens and ESBL-Producing Escherichia coli

Antibiotic resistance of microbial pathogens has become a threat to public health, with observed increase in outbreak of infections, therapeutic failure, morbidity and mortality. Pathogenic Escherichia coli and other extended spectrum beta lactamase (ESBL) producing bacteria causes serious health challenges due to antimicrobial resistance. This has caused an increasing research on medicinal plants as a source of alternative potential therapeutic agents. In this study, antimicrobial activity of Usteria guineensis (UG) and Sphaerocoryne gracilipes (SG) was investigated on standard organisms and clinical isolates of ESBL-producing E. coli. The leaves of Usteria guineensis and Sphaerocoryne gracilipes were extracted using methanol. Phytochemical analysis was carried out on the medicinal plants according to standard procedure. The clinical isolates of E. coli were screened for the production of ESBL using double disc synergy test, with E. coli ATCC 25922 as standard. Agar well diffusion method was used to determine antibacterial activity of the extracts at 100 and 25 mg/mL while Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were assessed using broth micro-dilution method. Statistical analysis was done using ANNOVA. The two plants contain cardiac glycoside, alkaloids and saponin and 11/15 (80%) of the E. coli isolates show the production of the ESBL. The extracts of the two plants showed antibacterial activity against ESBL E. coli with zone of inhibition ranging between 11 to 18 mm for Usteria guineensis and 12 to 15 mm for Sphaerocoryne gracilipes. The MIC of the extracts ranged between 0.78 to 50 mg/mL and MBC from 12.5 to >50 mg/mL for the ESBL producers and the standard strains. Therefore, the two plants, especially Usteria guineensis have potentials to be developed as alternative therapeutic agents for the treatment of infections caused by ESBL-producing pathogens like Escherichia coli.

The Etiology of Bloodstream Infections at an Italian Pediatric Tertiary Care Hospital: A 17-Year-Long Series.

Knowledge of epidemiology is essential for guiding correct antibiotic prescription, reducing bacteremia-associated mortality, and implementing targeted infection control programs. However, only a few studies have reported on the epidemiology of bloodstream infections (BSIs) in pediatrics. We performed a retrospective analysis of all BSIs (excluding those caused by common skin contaminants) diagnosed from 2006 to 2022 in patients younger than 18 years who were treated at an Italian pediatric tertiary care hospital. Overall, 2395 BSIs were recorded, including 2207 (92.15%) due to bacteria and 188 (7.85%) due to fungi. The incidence rate (BSIs/10,000 hospital discharges, IR) of bacterial BSIs significantly increased during the study period. In particular, BSIs caused by S. aureus (including MRSA), Enterobacterales (including ESBL and AmpC producers), Enterococcus spp., and P. aeruginosa became more common. The frequency of carbapenem-resistant strains was <1% and stable over time. Conversely, there was a significant reduction in the incidence of BSIs due to S. pneumoniae. The BSIs were stratified by patient age, and S. aureus was the most frequent cause of BSIs in all age groups, while E. coli was the most frequent in the Enterobacterales family. S. agalactiae was the third most frequent cause of neonatal early-onset BSIs. The prevalence of Enterococcus spp. increased in the subgroups from 8 days to 5 years of age, while P. aeruginosa became more prevalent in children over 5 years of age. S. aureus was also the most frequent isolate in both community- and hospital-onset BSIs, followed by E. coli. The prevalence of multidrug-resistant (MDR) pathogens was very low. It was <5% for both Gram-positive (i.e., MRSA and VRE) and Gram-negative (ESBL, AmpC, and carbapenem-resistant) pathogens, and MDR pathogens were almost exclusively detected in hospital-onset BSIs. Fungi accounted for just under 8% of BSIs. C. albicans was the most frequently isolated strain, followed by C. parapsilosis. Notably, the IR of fungemia did not change significantly during the study period, in spite of an increase in the absolute number of events. The continuous monitoring of local epidemiology is essential to identify changes in the IRs of pathogens and antibiotic susceptibility and to guide antibiotic treatments, especially in the phase when antibiograms are not yet available.

Antibacterial Resistance and Extended-Spectrum Beta-Lactamase (ESBL) Phenotypes in Enterobacteriaceae Isolated from Fecal Samples of Humans and Animals in Selected Local Government Areas of Nasarawa State, Nigeria

It is quite alarming the increasing rate of antibacterial resistance all over the world considering the public health threat and the re-emergence of multi-drug resistant Enterobacteriaceae. The aim of this study is Antibacterial resistance and phenotypic detection of Extended Spectrum Beta-Lactamase (ESBL) producing Enterobacteriaceae isolated from human and animal fecal samples in selected local government areas of Nasarawa state, Nigeria was carried out in the study. Hundred (100) samples comprising human and animal (goats, cattle, and chicken) were collected and 55 samples were multidrug resistant. A commercial biochemical kit (Eneterosystem 18R) was used for the isolation and identification of Enterobacteriaceae. Kirby Bauer Disk Diffusion Method was used for antibacterial susceptibility testing of Enterobacteriaceae isolates. The Double Disc Synergy Test (DDST) method was also used for the phenotypic confirmation test of Extended Spectrum Beta Lactamase (ESBL). The occurrence of multidrug-resistant isolates shows that Escherichia coli (100.00%) which is the highest, Proteus mirabilis (14.54%), Klebsiella pneumoniae, and Salmonella enterica (10.90%), while the occurrence of Shigella flexneri (9.09%) was the lowest. The Enterobacteriaceae isolates were more resistant to Cefuroxime, Cefexime, Amoxicillin Clavulanate, and Imipenem/Cilastatin with percentage resistance ranges from 66.6% - 100%. The occurrence of ESBL producers shows that Escherichia coli (60.00%) and Proteus mirabilis (62.5%) were high while Shigella flexneri (20.0%) had a low occurrence of ESBL. The sale and in-discriminate use of antibiotics without a prescription is an important regulatory issue in the abuse of antibiotics for both humans and animals. The Beta-Lactam and gentamycin antibiotics were not effective against the Multi-Drug Resistant (MDR) isolates and most of the isolates were ESBL producers.

Extended-spectrum beta-lactamase (ESBL) production and virulence genes profile of avian pathogenic Escherichia coli (APEC) isolated from broiler chickens in eastern Thailand

Avian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in extensive economic losses for the broiler industry. To date, there is little information in Thailand about virulence-associated genes and antibiotic resistance of APEC strains. Here, this study aimed to investigate the virulence genes and extended-spectrum beta-lactamase (ESBL) characteristics of APEC isolated from broilers. This study used multiplex polymerase chain reaction (PCR) to determine the presence of virulence genes and resistance genes in APEC. Furthermore, the disc diffusion method examined ESBL phenotypes of APEC and antibiotic resistance profiles against 17 antimicrobials. In this study, E. coli was isolated 11 (6.32%) of 174 broiler visceral organs. All E. coli isolates were tested for five APEC-virulence-associated genes (iroN, ompT, hlyF, iutA, and iss). Eight E. coli isolates from broilers with colibacillosis-associated lesions carried two (iroN and ompT) of the APEC virulence genes. One APEC virulence gene (hlyF) was found in E. coli isolates from broilers without lesions. Possibly no individual virulence gene was specific to the APEC strain. Interestingly, the papC, previously detected in humans with uropathogenic E. coli, was found in an APEC isolate. All APEC isolates were ESBL-producing E. coli, and then they were tested for four beta-lactamase-encoding genes (blaTEM, blaCTX-M, blaOXA, and blaSHV). The blaTEM and blaCTX-M were identified in 81.81% (9/11) of the isolates, whereas blaOXA or blaSHV were not detected in any isolate. All APEC showed multi-drug resistance (MDR) phenotypes, especially chloramphenicol, erythromycin, and sulfamethoxazole-trimethoprim. Although antibiotics were not recently used, MDR might be encouraged by horizontally transferring antibiotic-resistance genes. In addition, fluoroquinolone resistances were found in APEC isolates which could transfer resistance genes to humans via the food chain. This study indicates that APEC isolates contain several virulence and bla genes and should be surveillance to prevent the transmission of those genes to humans.

Trends in the Prevalence of Antimicrobial Resistance in Escherichia coli Isolated from Outpatient Urine Cultures in French Amazonia.

Antimicrobial resistance (AMR) in the community is increasing worldwide. We aimed to assess AMR trends in Escherichia coli from the community urine isolates in French Amazonia. We conducted a retrospective study from January 2016 to December 2022 in the Cayenne General Hospital microbiology laboratory (French Guiana). It included all urine samples positive for E. coli collected from adult outpatients. During the study period, 3,443 urinalyses positive for E. coli were studied. In 46% of cases, patients were women. In 64.4% of cases, E. coli were β-lactamase producers. The most frequently diagnosed resistance mechanisms were penicillinase production and sparing third-generation cephalosporins. Isolated E. coli were extended-spectrum β-lactamase (ESBL) producers in 6.1% of cases. Overall, E. coli was susceptible to amoxicillin in 35.9% [95% CI: 34.3-37.5], to amoxicillin/clavulanic acid in 62.2% [95% CI: 60.6-63.9], to cefotaxime in 94% [95% CI: 93.1-94.7], to gentamicin in 92.1% [95% CI: 89.1-92.6], to ofloxacin in 76.8% [95% CI: 75.3-78.2], to trimethoprim/sulfamethoxazole (SXT) in 58.8% [95% CI: 57.1-60.5], to fosfomycin in 99.1% [95% CI: 98.6-99.4], and to nitrofurantoin in 99% of cases [95% CI: 98.6-99.3]. We have observed a gradual decline in the susceptibility profile of E. coli for amoxicillin/clavulanic acid (P <0.001), piperacillin/tazobactam (P = 0.003), and temocillin (P = 0.006). However, susceptibility to ciprofloxacin was increasing (P = 0.001). In contrast, the susceptibility trends for amoxicillin, third-generation cephalosporins, gentamicin, SXT, nitrofurantoin, and fosfomycin remained stable over the 28 quarters of the study. In conclusion, isolated E. coli from outpatient urinalyses showed increased resistance profiles involving penicillinase and ESBL production. Close monitoring and strategies to decrease antibiotic consumption in the community are needed.

Phenotypic and Molecular Characterization of Escherichia coli Isolated from Food Animals for Antimicrobial Resistance

Background: The use of antimicrobials in food animals significantly contributes to the development of antimicrobial resistance in nature. This study aimed to determine the prevalence of ESBL-producing E. coli isolated from cattle, buffalo, sheep and goat fecal samples. Methods: In this study out of 2025 healthy animals from 10 different farms, fecal samples were randomly collected by rectal swabs from 200 animals (50 each from cattle, buffalo, sheep and goat). E. coli was isolated and further studied for AMR and ESBL production by phenotypic assays and PCR targeting ESBL genes. Result: The overall prevalence of E. coli was 81.50% with the highest isolation rate from cattle and goat samples. Out of 163 isolates, 45.39% were ESBL-producers. Genes encoding beta-lactam resistance viz. blaSHV, blaTEM, blaCTX-M and blaOXA were detected in 44.59% ESBL-positive isolates. The blaSHV was present in only cattle isolates, while blaCTX-M was present in E. coli isolated from all animals. The prevalence of beta-lactam genes was found as blaSHV (2.70%), blaTEM (12.16%), blaCTX-M (22.97%) and blaOXA (6.75%). A high degree of resistance to multiple antimicrobials was observed. Maximum susceptibility of E. coli was observed for trimethoprim, imipenem and chloramphenicol. This study demonstrated that food animals could be the source of ESBL-producing multidrug-resistant E. coli. The present findings are significant in the context of AMR monitoring in India.

Salmonella Prevalence and antibiotic resistance profile in raw poultry meat sold in North Lebanon: Insights from the COVID-19 pandemic and economic crisis

Salmonella-related foodborne illness is a significant public health concern, with the primary source of human infection being animal-based food products, particularly chicken meat. Lebanon is currently experiencing a dual crisis: the COVID-19 pandemic and an unprecedented economic crisis, which has resulted in substantial challenges to the public health system and food safety. This study aims to assess the prevalence and antibiotic resistance profile of Salmonella in raw poultry meat sold in North Lebanon during this dual crisis. A cross-sectional study was carried out between May 2021 and April 2022 across six different districts in North Lebanon. A total of 288 whole, unprocessed chickens were examined. The isolation and identification of Salmonella isolates were done based on cultural and biochemical properties. All isolates were subjected to antimicrobial susceptibility testing and phenotypic assays for Extended-Spectrum Beta-lactamase (ESBL) detection. The prevalence of Salmonella in raw poultry meat purchased in North Lebanon reached 18.05 % (52/288). The dry season and chilled chicken were significantly associated with an increased risk of Salmonella contamination (P < 0.05). Additionally, 34.61 % of the isolates were potential ESBL producers, and 57.69 % exhibited multidrug resistance (MDR). This study highlights the existence of MDR in chicken meat in North Lebanon, posing a potential health risk if undercooked chicken meat is consumed. This emphasizes the importance of the implementation of preventive strategies and hygienic procedures throughout the food chain to reduce the risk of Salmonella spp. contamination in chicken meats and its potential transmission to humans.

Molecular Characteristics of Cephalosporin Resistant Escherichia coli and Klebsiella pneumoniae Isolated from Children in a Tertiary Care Centre of Central Kerala, India

Aims: The study was aimed to determine the molecular characteristics of extended-spectrum beta-lactamases (ESBL) producing cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae isolated from children below ten years of age. Background: Geographically diverse variations in the prevalence of ESBL genes were reported. No data were available on the prevalence of ESBL genes in central Kerala, India, among children below 10 years of age. Methods: A cross-sectional study was performed to analyze ESBL genes in cephalosporin-re-sistant E. coli and K. pneumoniae strains isolated from samples received in the Microbiology la-boratory of a tertiary care centre during the period between May 2021 and July 2022. The strains showed that ESBL + cephalosporin resistance was subjected to PCR-based genotyping for the genes such as bla (beta-lactamase) CTX-M-1, blaCTX-M-15, blaCTX-M-U, blaTEM, blaPER and SHV. Results: Among the total 228 samples analyzed, 136 (60%) had no growth. Ninety-two (40 %) samples showed growth of E. coli and K. pneumoniae. Among the isolates that showed growth, 39 (42%) were sensitive, and the remaining 53 (57%) were resistant to third-generation cephalospor-ins. Among the isolates showed resistance, 22 (42%) were ESBL positive and 31 (58%) were ESBL negative. Among the positive ESBL, nine E. coli strains (60%) were positive for CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were present in six (85%) K. pneumoniae with ESBL +. Conclusion: E. coli and K. pneumoniae isolated from specimens of children below ten years of age showed 41-42% ESBL producers. Prevalent ESBL-producing genes in E. coli were CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were prevalent in ESBL-producing K. pneumoniae.

Investigation of plasmid-mediated quinolone resistance among extended-spectrum β-lactamase isolates of E. coli and K. pneumoniae.

Escherichia coli and Klebsiella pneumoniae are important members of the Enterobacteriaceae family, involved in many infections. The increased resistance rate towards β-lactams and fluoroquinolones -which are the main therapeutic options- limits their treatment options. This study aimed to assess the local resistance patterns against different antimicrobials and to determine the extended-spectrum β-lactamase (ESBLs) producers. The study revealed that 36% of clinical isolates were ESBL producers, showing high resistance rates towards β-lactams and non-β-lactams, especially sulphamethoxazole-trimethoprim and fluoroquinolones. However, they were susceptible to chloramphenicol and doxycycline (33% and 20%; respectively). Also, the investigation aimed to screen the plasmid profile of quinolone-resistant ESBLs-producers and to detect the plasmid-mediated quinolone resistance genes including qnrA, qnrS, qnrB, qnrC, qnrD, and qnrVC. Moreover, the conjugative plasmid among the quinolone-resistant isolates was elucidated. The results showed that extracted plasmids of sizes ranging from ≈0.9 to 21.23 Kb, divided into 7 plasmid patterns were detected. A plasmid of approximately 21.23 Kb was found in all isolates and the QnrS gene was the most predominant gene. Moreover, the frequency of transconjugation within the same genus was higher than that recorded between different genera; where 68% of E. coli isolates transferred the resistance genes compared to Klebsiella isolates (36.6%). Plasmid profiles of transconjugants demonstrated great similarity, where 21.23 Kb plasmid was detected in all transconjugants. Since these transconjugants were quinolone-resistant ESBL producers, it has been suggested that quinolone resistance determinants might be carried on that plasmid.

Sensitivity and specificity of Nanopore sequencing for detecting carbapenem and 3rd-generation cephalosporin-resistant Enterobacteriaceae in urine samples: Real-time simulation with public antimicrobial resistance gene database

ObjectivesTo evaluate the accuracy of beta-lactamase gene detection directly from urine samples by Nanopore sequencing. MethodsDNA was extracted from bacterial pellets in spun urine. The purified DNA was then sequenced in native form by a Nanopore sequencer (MinION) to identify the organisms and beta-lactamase genes. Results were compared to routine urine cultures and standard antimicrobial susceptibility tests (AST). ResultsWe processed 60 urine samples of which routine cultures grew Enterobacteriaceae, including 28 carbapenem-resistant (CRE), 17 extended-spectrum beta-lactamase (ESBL) or AmpC producing, and 15 non-ESBL/AmpC phenotypes. We excluded 7 samples with extremely low DNA amounts (<1 ng/μl) for a final case of 53 in total. The sensitivity of antimicrobial resistance gene detection within 6 h, the optimal duration from real-time simulation, of Nanopore sequencing for the diagnosis of carbapenem-resistant and ceftriaxone-resistant phenotypes was 73.9 % (95%CI 56.0–91.9 %) and 81.1 % (95%CI 68.5–93.7 %), while the specificity was 96.7 % (95%CI 90.2–100.0 %) and 56.3 % (95%CI 31.9–80.6 %), respectively. The median times for MinION to generate DNA reads containing carbapenemase and ESBL/AmpC genes were 93 min (IQR 17–245.5) and 99 min (IQR 31.25–269.75) after sequencing commencement, respectively. ConclusionsNanopore sequencing can identify bacterial genotypic resistance in urine and may enable clinicians to adjust antimicrobial therapy earlier than routine AST.

Antibiotic Resistance Profiles of Extended-Spectrum β-Lactamase (ESBL)- and Metallo-β-Lactamase (MBL)-Producing Klebsiella pneumoniae Isolates From Diabetic Foot Ulcers: Implications for Treatment Strategies.

Background Diabetic foot ulcers (DFUs) are prevalent complications of diabetes mellitus, often leading to severe infections and adverse clinical outcomes. Klebsiella pneumoniae, a gram-negative bacterium, has emerged as a significant causative agent in DFU infections, raising concerns due to its increasing antibiotic resistance, particularly in extended-spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) production. Aim This study aimed to comprehensively assess the prevalence, antibiotic resistance profiles, and clinical correlates of ESBL- and MBL-producing K. pneumoniae isolates specifically derived from DFUs. Methods A cross-sectional observational study was conducted at Krishna Vishwa Vidyapeeth from January 2023 to June 2023, involving 126 patients diagnosed with DFUs. Clinical and demographic data were collected, and wound swabs underwent microbiological analysis. Phenotypic detection methods were employed to identify ESBL and MBL production, followed by standardized antibiotic susceptibility testing. Results Among the 126 isolates tested, 36 (28.6%) were identified as ESBL-producing and 21 (16.7%) as MBL-producing strains. ESBL-producing isolates exhibited high resistance rates to antibiotics such as ampicillin (92.3%), amoxicillin-acid (84.6%), and cephalosporins, including ceftriaxone (76.9%), and cefepime (73.8%). MBL-producing isolates demonstrated even broader resistance profiles, including resistance to fluoroquinolones (ciprofloxacin, 60.0%; levofloxacin, 57.1%), aminoglycosides (gentamicin, 42.9%), and carbapenems (meropenem, 38.1%; imipenem, 35.7%). Conclusion This study identifies a significant prevalence of ESBL- and MBL-producing K. pneumoniae in DFUs, showcasing high antibiotic resistance rates. Comorbidities correlate significantly with the presence of resistant isolates, necessitating treatment strategies for effective management.

Monitoring Multi-Drug Resistant Klebsiella pneumoniae in Kitagata Hot Spring, Southwestern Uganda: A Public Health Implication.

The concerning frequency of K. pneumoniae in various recreational settings, is noteworthy, especially regarding multi-drug resistant (MDR) strains. This superbug is linked to the rapid spread of plasmids carrying these resistance genes. The objective of this study was to evaluate the spatiotemporal prevalence of MDR-K. pneumoniae in the Kitagata hot spring, Southwestern Uganda. A laboratory-based descriptive longitudinal study was conducted between May and July 2023. During rainy and dry seasons, we collected eighty water samples in the morning and evening from the hot spring. The temperature at each point was measured prior to sample collection, and two samples were obtained at varying depths. 5 mL of each homogenized sample were pre-enriched in brain heart infusion broth, and subsequently in both blood and violet red bile agar. The Kirby-Bauer disk diffusion method was performed, followed by the detection of carbapenemase (CR) and extended-spectrum β-lactamase (ESBL) production. Polymerase chain reaction showed resistance genes viz. bla TEM, bla CTX-M and bla KPC. Data were analyzed using SPSS-20 to obtain chi-square tests and regression analysis. K. pneumoniae accounted for 30.0% of isolates obtained from Kitagata hot springs, with all isolates classified as multi-drug resistant. All isolates were resistant to ampicillin, rifampicin, ceftazidime, and azithromycin (79.2%). Additionally, 95.8% of isolates harbored bla TEM gene alone and both bla TEM and bla CTX genes, followed by bla KPC alone (33.3%), with 25% harboring all three resistance genes. During the dry season, K. pneumoniae had a higher prevalence (35.0%) compared to the wet season (25.0%). The prevalence of MDR-K. pneumoniae significantly increased over the course of the study. The presence of the three studied resistance genes in the isolates showed a positive correlation with the second phase of sample collection and the dry season but exhibited a negative correlation with temperature, except for isolates harboring either bla TEM alone or bla TEM+KPC+CTX genes. Kitagata hot spring serves as a hotspot for continuous dissemination and acquisition of MDR-K. pneumoniae harboring resistance genes that encode for ESBL and CR production. The healthcare sector ought to implement an ongoing monitoring and surveillance system as well as robust antimicrobial resistance stewardship programs aimed at delivering health education to the community.

Assessment of in vitro activity of ceftazidime/avibactam on carbapenemase-producing Enterobacterales from Iran: An experimental study.

The prevalence of carbapenemase-producing Enterobacterales (CPE) continues to increase worldwide. Combination of β-lactam and novel β-lactamase inhibitors introduce a revolutionary treatment option for CPE. Ceftazidime/avibactam (CAZ/AVB) has been recently developed for treatment of severe infections caused by multidrug-resistant bacteria. We aimed to evaluate in vitro activity of CAZ/AVB on a collection of 85 ESBL-producing-carbapenemase negative and CPE from Iran. ESBL and carbapenemase production was phenotypically confirmed by combined disk test and modified carbapenem inactivation method respectively. The presence of clinically important carbapenemase encoding genes was examined using PCR. Susceptibility of all isolates to CAZ/AVB was determined using discs containing 30 μg ceftazidime +20 μg avibactam (AVB). Minimum inhibitory concentrations (MICs) of CAZ/AVB in 28 CPE (4 Escherichia coli and 24 Klebsiella pneumoniae) was determined by gradient diffusion method using MIC test strips (0.016-256 mg/L ceftazidime +4 mg/L AVB). All phenotypically identified ESBL positive-carbapenemase negative isolates were found to be susceptible to CAZ/AVB. Among the carbapenem resistant isolates, CAZ/AVB showed potent inhibitory activity against all OXA-48-like (MIC ranges 0.125/4-0.75/4 mg/L) and KPC positive isolates (MIC ranges <0.016/4-0.19/4 mg/L). However, AVB could not restore the activity of ceftazdime against isolates producing metallo-β-lactamases (MLBs) including VIM, NDM (MIC > 256/4 mg/L) and IMP (MIC > 8/4 mg/L). Our data highlighted the excellent in vitro performance of CAZ/AVB against ESBL-producing and CPE suggesting that this combination can efficiently be used as therapeutic option for management of CPE infections particularly in regions with high prevalence of KPC and/or OXA-48-like positive but MBL-negative Enterobacterales.

Extended-spectrum beta-lactamase and carbapenemase producing Klebsiella Pneumoniae isolated from patient with suspected urinary tract infection

Urinary tract infection (UTI) is the most common bacterial infections in humans and serious health problem in many parts of the world. Carbapenem resistant Enterobacteriaceae (CRE) are increasingly reported from healthcare facilities in Nepal and around the world. This study is carried out with an objective to assess the Prevalence of ESBL and carbapenemase producing Klebsiella pneumoniae isolate from patient with suspected of UTI. Total 880 urine sample was collected during 6-month period and processed and identified by using conventional microbiological procedure and biochemical test. ESBL screening isolates were done by using Ceftazidime and Ceftazidime with clavulanic acid. Production of ESBL was determine by combination Disc Test. For the confirmation of carbapenemase producing K. pneumoniae Modified Hodge Method, Carbapenem Inactivation Method and EDTA Impregnated Disc test (MBL) was performed. Out of total sample, 9.30% (82) showed significant growth. Prevalence of UTI seen more in male as compare to female patients. Out of positive isolates, 37.8% were K. pneumoniae. K. pneumoniae were resistance with Ceftriaxone (54.8%) followed by 51.61% Ceftazidime, Cotrimoxazole. Multi drug resistance (MDR) was found to be 54.83%. Out of 31 isolates, 10(32%) were confirmed as ESBL positive by Combination Disc Assay. 14 isolates were screened as carbapenemase producers but 2(14.28%) showed positive through Modified Hodge Method, 4(28.57%) Modified Carbapenem Inactivation Method and 7(50%) EDTA impregnated Disc Test (MBL). Analytically, the sensitivity of MHT, CIM, and MBL tests was 42.81%, 58.10%, and 76.96%, respectively, all with 100% specificity. Colistin was the drug of choice, with 93.54% of isolates showing sensitivity to it. Early detection of ESBL and MBL-producing isolates is crucial for reducing mortality rates and preventing the intra-hospital spread of these strains.

Acute Respiratory Distress Syndrome in a Patient with Myasthenia Gravis and Septic Shock: A Case Report

Introduction: Myasthenia gravis (MG) is an autoimmune disorder characterized by muscle weakness, often affecting respiratory and oropharyngeal muscles. This predisposition to respiratory compromise, coupled with impaired swallowing and the potential need for immunosuppressive therapies, increases the risk of pneumonia and subsequent sepsis in MG patients. Sepsis, in turn, is a significant risk factor for acute respiratory distress syndrome (ARDS), a severe lung condition with high mortality. Case presentation: We present the case of a 47-year-old male with a 4-year history of MG who was admitted to our hospital with progressive dyspnea and dysphagia. His condition deteriorated rapidly, leading to septic shock and respiratory failure necessitating invasive mechanical ventilation. Blood cultures identified Klebsiella pneumoniae with extended-spectrum beta-lactamase (ESBL) production. Despite aggressive treatment, including therapeutic plasma exchange (TPE), the patient's hospital course was complicated. Conclusion: This case underscores the critical importance of vigilant monitoring and early intervention in MG patients presenting with respiratory symptoms or signs of infection. Prompt recognition and aggressive management of sepsis are crucial to mitigate the risk of ARDS and improve outcomes in this vulnerable patient population.

Prevalence of Extended-Spectrum ß-Lactamases (ESBLs) and AmpC ß-Lactamases in Clinical Isolates of Multiple Drug-Resistant Escherichia coli

The current study aims to investigate the prevalence of extended-spectrum ß-lactamases (ESBLs) and AmpC β-lactamases in multiple-drug-resistant Escherichia coli (e.g., multidrug-resistant (MDR), extensively drug-resistant (XDR), and pan-drug-resistant (PDR)) collected from hospitalized patients in Baghdad. The results showed that the prevalence of ESBLs among E.coli isolates was high. From the total 113 E.coli isolates, 75 (66.37%) were ESBL-producing, while 38 (33.63%) were non-ESBL-producing. Out of 75 ESBL-positive isolates, 39 (52%) were obtained from females and 36 (48%) from males. Additionally, 43 (57.33%) isolates were collected from urine samples, and the highest production of ESBLs was obtained from the age group 41–60 years (29.33%). Moreover, out of 111 MDR E.coli, 64 (57.66%) exhibited a positive ESBL test, while 47 (42.34%) did not. Out of 24 XDR E.coli isolates, 11 (45.83%) demonstrated positive ESBLs, while 13 (54.17%) showed negative ESBLs. The antimicrobial susceptibility test results showed that positive ESBL E.coli isolates were more drug-resistant than negative ESBL isolates. The positive ESBLs of E.coli exhibited a higher resistance rate to the β-lactam antibiotics and showed a co-resistance to non-β-lactam antibiotics. Phenotypic detection of AmpC β-lactamase by the screening of cefoxitin-resistant isolates revealed that 43 (38.05%) isolates were considered positive for AmpC β-lactamase production. However, the PCR technique gives different results. In conclusion, the prevalence of ESBL and AmpC β-lactamase producing E. coli is rapidly increasing among clinical isolates of MDR, XDR, and possibly PDR E.coli.

Bacterial diversity in Buruli ulcer lesions in Ghana

BackgroundPrevious studies have demonstrated secondary microbial infection of Buruli ulcer (BUD) lesions before, during and after treatment. However, there is limited data on the bacterial diversity across treatment and their influence on clinical outcome. The present study aimed to investigate the relationship between bacterial diversity within BUD lesions and clinical outcome in affected individuals. MethodsWe investigated the bacterial diversity within lesions of individuals with PCR confirmed BUD from 5 endemic districts within central Ghana. Samples were collected longitudinally from lesions over treatment period. Microbiological analyses including isolation of bacteria, and species identification were performed using the VITEK 2 compact. ResultsOut of 36 participants included, 80.5 % presented with ulcers on the lower limbs. Higher bacterial diversity was observed in ulcers compared to other clinical forms of BUD. There was a significant association between bacterial diversity and clinical outcome (p = 0.002). ESBL producing bacteria and MRSA were isolated in slow healing BUD lesions. ConclusionHigher diversity of secondary organisms colonizing BUD lesions may have an impact on clinical outcome in affected individuals. There is the need for the development of treatment guidelines for simultaneous management of M. ulcerans and other potential pathogens within lesions to improve clinical outcome.

Analysis of antibiotic resistance in Gram-negative bacilli in wild and exotic healthy birds in Brazil: A warning sign

Bacterial antibiotic resistance is a public health problem affecting humans and animals. This study focuses on identifying Gram-negative bacilli (GNB) (MALDI-TOF MS and Klebsiella MALDI TypeR) resistant to antimicrobials in freshly emitted feces of healthy captive and rescued wild birds from a zoo in Brazil. Birds from the zoo and rescued from sixteen different orders were investigated. Resistant bacteria from feces were selected (MacConkey agar with 2 μg/mL cefotaxime). Genomic similarity and plasmid were investigated by Pulsed-Field Gel Electrophoresis of XbaI fragments (XbaI-PFGE) and S1-PFGE. Polymerase Chain Reaction (PCR) was performed to search for beta-lactamase genes. From 80 birds included, 26 from the zoo (50 %) and 18 rescued wild birds (64 %) presented cefotaxime-resistant GNB. E. coli and Klebsiella spp were the most prevalent species. Among 65 isolates from the zoo and rescued wild birds, 75 % were considered multidrug-resistant (MDR). The majority of the isolates were extended-spectrum beta-lactamases (ESBL) producing and resistant to enrofloxacin. blaCTX-M-GROUP-1, blaTEM, and blaSHV were the most detected genes, and blaKPC was detected in K. pneumoniae complex. According to genomic similarity results, some identical profiles were found in birds with no known contact among the zoo or rescued birds. Several isolates carried one to three plasmids (15–350 kb). The presence of multidrug-resistant (MDR) isolates from healthy captive and wild birds brings novel data on the dissemination of these elements to the environment.

Antibiotic resistance and its correlation with biofilm formation and virulence genes in Klebsiella pneumoniae isolated from wounds.

Klebsiella pneumoniae 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.

Efficacy of Nitrofurantoin and Fosfomycin against Extended Spectrum Beta Lactamase Producing Uropathogens in the Era of Antibiotic Resistance: A Cross-sectional Study

Background: Urinary Tract Infection (UTI) is one of the most common bacterial infections that require medical care. It accounts for 25% of all infections. Limited number of antibiotics are available for the treatment of UTI due to increased resistance among uropathogens. Aim of the study is to determine the causative agents of UTI and to evaluate the action of nitrofurantoin and fosfomycin against Enterobacteriaceae in ESBL (Extended spectrum beta-lactamase) producing strains. Material and Methods: It is a retrospective study carried out in the department of Microbiology, MAMC, Agroha. Data related to urine culture and antibiotic susceptibility pattern from January 2023 to December 2023 was taken from the records and analysed. Results: Out of 1648 urine samples processed, 569 showed significant bacterial growth. Escherichia coli (45.3%) was the most common isolate followed by Klebsiella pneumoniae (20.5%), Enterococcus (17.2%), Staphylococcus aureus (5%). Among the gram negative bacilli, 40% were ESBL producers. These ESBL producing strains were 99% sensitive to Fosfomycin and 75% sensitive to Nitrofurantoin. Conclusion: The study revealed that Nitrofurantoin and Fosfomycin are very effective oral drugs for empirical treatment against gram negative uropathogens causing UTI, especially multi drug resistant strains.