Extended-spectrum Beta-lactamase Isolates Research Articles

Evaluation of virulence determinants and cell surface properties associated with biofilm formation in methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum beta-lactamase (ESBL) Escherichia coli from livestock and poultry origin

Antibiotic resistance poses a persistent threat to modern medicine due to the emergence of novel antibiotic-resistant strains. Therefore, a timely understanding of antibiotic resistance and the virulence biology of pathogenic bacteria, particularly those of public health significance, is crucial for implementing effective mitigation strategies. This study aimed to investigate the virulence profiles of ten S. aureus isolates (NDa to NDj) and ten E. coli isolates (ND1 to ND10) originating from livestock and poultry, and to assess how various cell surface properties and biofilm formation abilities influence antibiotic resistance phenotypes. Antibiotic resistance profiling through phenotypic (AST) and genotypic methods (PCR) confirmed that NDa to NDe were methicillin-resistant S. aureus (MRSA) and ND1 to ND5 were extended-spectrum β-lactamase (ESBL) producing E. coli isolates. Virulence properties such as hemolytic activity, coagulase activity, and nuclease activity were found to be independent of the antibiotic resistance phenotype in S. aureus. In contrast, biofilm formation phenotype was observed to influence antibiotic resistance phenotypes, with MRSA and ESBL E. coli isolates demonstrating higher biofilm formation potency. Chemical and enzymatic analysis of S. aureus and E. coli biofilms revealed proteins and polysaccharides as major components, followed by nucleic acids. Furthermore, cell surface properties such as auto-aggregation and hydrophobicity were notably higher in isolates with strong to medium biofilm-forming capabilities (ESBL and MRSA isolates), corroborated by genomic confirmation of various genes associated with biofilm, adhesion, and colonization. In conclusion, this study highlights that surface hydrophobicity and biofilm formation ability of MRSA (NDa to NDe) and ESBL E. coli (ND1 to ND5) isolates may influence antibiotic resistance phenotypes.

Colistin resistance in ESBL- and Carbapenemase-producing Escherichia coli and Klebsiella pneumoniae clinical isolates in Cambodia

Despite the critical importance of colistin as a last-resort antibiotic, limited studies have investigated colistin resistance in human infections in Cambodia. This study aimed to investigate the colistin resistance and its molecular determinants among Extended-spectrum beta-lactamase (ESBL)- and carbapenemase-producing (CP) Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) isolated in Cambodia between 2016 and 2020. E. coli (n = 223) and K. pneumoniae (n = 39) were tested for colistin minimum inhibitory concentration (MIC) by broth microdilution. Resistant isolates were subjected to polymerase chain reaction (PCR) for detection of mobile colistin resistance genes (mcr) and chromosomal mutations in the two-component system (TCS). Eighteen isolates (10 K. pneumoniae and 8 E. coli) revealed colistin resistance with a rate of 5.9% in E. coli and 34.8% in K. pneumoniae among ESBL isolates, and 1% in E. coli and 12.5% in K. pneumoniae among CP isolates. The resistance was associated with mcr variants (13/18 isolates, mcr-1, mcr-3, and mcr-8.2) and TCS mutations within E. coli and K. pneumoniae, with the first detection of mcr-8.2 in Cambodia, the discovery of new mutations potentially associated to colistin resistance in the TCS of E. coli (PhoP I47V, PhoQ N352K, PmrB G19R, and PmrD G85R) and the co-occurrence of mcr genes and colistin resistance conferring TCS mutations in 11 of 18 isolates. The findings highlight the presence of colistin resistance in ESBL- and CP- Enterobacteriaceae involved in human infections in Cambodia as well as chromosomal mutations in TCS and the emergence of mcr-8.2 in E. coli and K. pneumoniae. It underscores the need for continuous surveillance, antimicrobial stewardship, and control measures to mitigate the spread of colistin resistance.

Clinical, phenotypic, and genotypic characteristics of ESBL-producing Salmonella enterica bloodstream infections from Qatar

BackgroundResistant Salmonella infections are a major global public health challenge particularly for multidrug-resistant (MDR) isolates manifesting as bloodstream infections (BSIs). ObjectivesTo evaluate clinical, phenotypic, and genotypic characteristics of extended-spectrum beta-lactamase (ESBL) producing Salmonella enterica BSIs from Qatar. MethodsPhenotypic ESBL Salmonella enterica from adult patients presenting with positive BSIs were collected between January 2019 to May 2020. Microbiological identification and characterization were performed using standard methods while genetic characteristics were examined through whole genome sequencing studies. ResultsOf 151 episodes of Salmonella enterica BSI, 15 (10%) phenotypic ESBL isolates were collected. Recent travel was recorded in most cases (80%) with recent exposure to antimicrobials (27%). High-level resistance to quinolines, aminoglycosides, and cephalosporins was recorded (80-100%) while meropenem, tigecycline and colistin demonstrated universal susceptibility. Genomic evaluation demonstrated dominance of serotype Salmonella Typhi sequence type 1 (93%) while antimicrobial resistance genes revealed dominance of aminoglycoside resistance (100%), qnrS1 quinolones resistance (80%), blaCTX-M-15 ESBLs (86.7%), and paucity of AmpC resistance genes (6.7%). ConclusionsInvasive MDR Salmonella enterica is mainly imported, connected to patients from high prevalent regions with recent travel and antimicrobial use caused by specific resistant clones. In suspected cases of multidrug resistance, carbapenem therapy is recommended.

Multidrug-Resistant Extended-Spectrum Beta-Lactamase (ESBL)-Producing Escherichia coli in a Dairy Herd: Distribution and Antimicrobial Resistance Profiles.

This study investigated the presence, distribution, and antimicrobial resistance profiles of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in a dairy herd located in Northern Italy. The feces of clinically healthy calves, their mothers, and the cows treated for mastitis, as well as water, environmental samples, and waste milk were collected and subjected to bacteriological culture on CHROMagarTM ESBL plates. A questionnaire was administered to identify risk factors. The isolates were identified as E. coli by MALDI-TOF MS and subjected to the double-disk synergy test (DDST) and minimal inhibitory concentration (MIC) assay. As a result, ESBL E. coli was isolated from the feces of 28 of 37 (75.67%) calves, the feces of 2 of 3 (66.67%) treated cows, 8 of 14 (57.15%) environmental samples, and waste milk. All ESBL isolates showed multiple resistances and were categorized as multidrug-resistant (MDR). Several risk factors for ESBL E. coli selection and diffusion were identified, including lack of routine cleaning of calf feeding and housing equipment, administration of waste milk to male calves, and blanket dry cow therapy. In conclusion, this study highlighted the presence of MDR, ESBL E. coli in the feces of most dairy calves, and their association with different sample sources. Accordingly, adding to the prudent use of antibiotics, the adoption of adequate farm hygiene and biosecurity measures might also help prevent the spread and transmission of ESBL E. coli within the herd.

Antibiotic resistance among Aerobic Gram-Negative Bacilli isolated from patients with oral inflammatory dysbiotic conditions—a retrospective study

IntroductionAerobic Gram-Negative Bacilli (AGNB) are not part of the resident oral microflora but are occasionally found in high abundance under inflammatory dysbiotic conditions at various oral niches. The aim of the present study was to investigate the identity and antibiotic susceptibility of AGNB isolated from patients in Sweden with mucosal lesions, periodontitis, and peri-implantitis, with special attention to antibiotic resistance and on the presence of phenotypic Extended Spectrum Beta-Lactamase (ESBL) isolates.Materials and methodsMicrobiolgical samples were harvested from 211 patients in total, experiencing mucosal lesions (N = 113), periodontitis (N = 62), or peri-implantitis (N = 36). The growth of AGNB's was semiquantified by selective and non-selective culture and the strains were isolated, identified, and tested for antibiotic susceptibility. A total of 251 AGNB strains, occurring in moderate to heavy growth (>100 CFU/ml sample), indicating a dysbiotic microbiota, were identified. The disc diffusion method was used for screening of the antibiotic susceptibility of the isolates. Phenotypic identification of ESBL isolates was based on resistance to ceftazidime and/or cefotaxime.ResultsThe most commonly detected AGNB isolates in oral inflammatory dysbiotic conditions were fermentative species belonging to Enterobacteriaceae e.g. Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp, and the non-fermentative environmental Burkholderia cepacia, Pseudomonas spp., and Stenotrophomonas maltophilia. No clear trends were seen in frequency of the various species in samples from mucosal lesions, severe periodontitis, and peri-implantitis cases. The 138 Enterobacteriaceae isolates and 113 environmental AGNB isolated showed a high antibiotic resistance in general against antibiotics commonly used in dentistry (Amoxicillin, Amoxicillin + Clavulanic acid, Ampicillin, Clindamycin, Doxycycline, Erythromycin, Oxacillin, PenicillinV, and Tetracycline). The majority of these isolates were susceptible to ciprofloxacin. Ten isolates (4.1%) were phenotypically classified as ESBL positive. The ESBL isolates were predominantly found among isolates of S. maltophilia, while only one ESBL positive isolate was found among Enterobacteriaceae.ConclusionsPhenotypically identified ESBL isolates can occasionally be present among oral AGNB strains isolated in abundance from the dysbiotic microbiota occurring in cases with oral mucosal lesions, severe periodontitis, or peri-implantitis.

Prevalence of drug resistant Enterobacteriaceae in a Nepalese tertiary care hospital.

Antimicrobial resistance in Enterobacteriaceae is an emerging global public health problem. Numerous studies have reported community-acquired AmpC beta-lactamase and extended spectrum beta-lactamase (ESBL) producing Enterobacteriaceae in Nepal. However, there are limited data on community-acquired Metallo-beta-lactamase (MBL) producing Enterobacteriaceae. A hospital-based descriptive cross-sectional study was conducted using 294 Enterobacteriaceae isolates from a total of 2,345 different clinical specimens collected from patients attending a tertiary care hospital in Nepal. Bacteria were isolated using standard microbiological growth media and identified using biochemical tests. For antimicrobial susceptibility testing, Kirby-Bauer disc diffusion technique was used. AmpC, ESBL, and MBL productions were detected by using combined disc method. AmpC, ESBL, and MBL productions were detected in 19.4%, 29.6%, and 8.5% of total Enterobacteriaceae isolates respectively. Higher rates of beta-lactamases production were seen among the isolates from in-patients in comparison with those from out-patients. However, 11.6%, 25%, and 3.7% of the total isolates from out-patients were AmpC, ESBL, and MBL producers respectively. The co-production of the beta-lactamases was also detected, with two Klebsiella pneumoniae isolates producing all three beta-lactamases. One MBL producing Proteus vulgaris isolate that was pan-resistant with no remaining treatment options was also isolated. Prevalence of drug resistant Enterobacteriaceae in our study was very high. Detection of AmpC, ESBL, and MBL positive isolates from out-patients, who did not have recent history of hospital visit, indicated the community dissemination of the drug resistant bacteria. This is a matter of great concern and an immediate attention to formulate strategies to prevent further development and spread of antibiotic resistance is required.

Phenotypic and genotypic characterization of extended spectrum beta-lactamase producing E. coli harboring carbapenem and colistin-resistant genes from poultry farms in Egypt.

eEscherichia coli (E. coli) bacteria that produce extended spectrum beta-lactamase (ESBL) is associated with a high prevalence of human illnesses worldwide. The emergence of resistance to carbapenem and colistin compounds poses further challenges to the treatment options for these illnesses. This study aimed to evaluate the phenotypic and genotypic pattern of resistance to carbapenem and colistin in ESBL-producing E. coli. Escherichia coli isolates collected from the respiratory tract of chickens in El-Sharkia government, Egypt. A total of 250 lung samples were collected from 50 poultry farms. These samples were then subjected to isolation, identification, and serotyping of E. coli. The presence of antimicrobial resistance was identified by disc diffusion testing. The occurrence of ESBL phenotypes was also assessed using the double disc synergy method. PCR/sequencing techniques were employed to examine the presence of ESBL (β-lactamase (bla)-TEM, blaSHV, and blaCTX-M), colistin (mcr-1), and carbapenem (blaNDM, blaVIM, and blaKPC) resistance genes. The findings revealed that 140 out of 250 (56%) were identified as E. coli. All E. coli isolates had a high level of multi-antimicrobial resistance (MAR) with an index value greater than 0.2, and 65.7% of them were confirmed to produce ESBL. Out of the 92 ESBL phenotypes, 55 (59.7%), 32 (34.7%), 18 (19.6%), and 37 (40.2%) isolates harbor b laTEM-3, b laSHV-4, b laCTX-M-1, a nd blaCTX-M-14 genes, respectively. The blaNDM-1 gene was identified in all 40 phenotypes that exhibited resistance to carbapenem, accounting for 28.5% of all strains of E. coli and 43.4% of ESBL isolates. The VIM and KPC genes were not detected in any of the samples. Furthermore, there was a significant prevalence of the mobilized colistin resistance (mcr)-1 gene, with 64 (69.5%) of the ESBL isolates exhibiting this gene. The prevalence of ESBL-producing E. coli, particularly those resistant to carbapenem and colistin, poses a significant public health risk in society.

Prevalence of extended-spectrum beta-lactamases producing isolates obtained from patients of pediatric critical care unit in a tertiary care hospital

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.

Frequency of Extended-Spectrum Beta-Lactamases-Producing Escherichia coli Among Out- and In-patients in Rafsanjan City, Iran

Background: The frequency of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains among clinical isolates has been steadily increasing, leading to limited treatment options. Objectives: This study aimed to assess the antibiotic sensitivity of E. coli and the frequency of ESBL isolates among both out-patients and in-patients. Methods: A total of 390 E. coli isolates were received at the Ali-Ebn-e-Abitaleb Hospital laboratory in Rafsanjan. The antibiogram, as well as the phenotypic and genotypic detection of ESBL isolates, were conducted using Kirby-Bauer, combination disk confirmatory, and polymerase chain reaction tests, respectively. Results: Of all the E. coli isolates, 45.6% exhibited ESBL production. Among these isolates, 46.1% were obtained from hospital wards, while 42.5% were from outpatients. Meropenem and imipenem displayed sensitivities of 97.2% and 93.3%, respectively, whereas amikacin and nitrofurantoin showed sensitivities of 89.7% and 85.6%, respectively, for all isolates. Notably, ceftriaxone, cefotaxime, cefixime, ampicillin, co-trimoxazole, and nalidixic acid demonstrated high resistance rates, surpassing 50%. ESBL-producing isolates were more frequently observed in blood samples (65%) and wounds (60%) compared to other tested isolates. Approximately 8.6% of isolates carried a single type of ESBL gene, while 38.5% carried multiple ESBL genes. Conclusions: The data indicate a prevalence of ESBL-producing E. coli isolates among both out-patients and in-patients, with some of them acquiring two or three types of ESBL enzymes. As a result, their ability to hydrolyze antibiotics has increased, leading to their higher occurrence in clinical samples.

Plasmid and Total Protein Analyses of Extended Spectrum Beta Lactamase Producing Bacteria from Periplaneta americana (Coackroaches)

Background: The importance of plasmid-encoded Extended Spectrum Beta Lactamase (ESBL) organisms in the dissemination of multi-drug resistance and complicated infections are contributory factors to treatment failure. Despite these, there is a paucity of information regarding plasmid-borne associated infection from primary sources including cockroaches while studies correlating total protein and plasmid-encoded ESBL are also limited especially in cockroaches. The objective of this study was therefore aimed at determining the presence of plasmid in the ESBL organisms from cockroaches as well as their total protein profiles.
 Materials and Methods: Bacterial isolation, characterization and molecular identification were carried out in our previous work following standard recommended techniques. These molecularly identified organisms which were delineated into twenty-two (22) were further subjected to plasmid and total protein analyses using alkaline lysis and Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) respectively.
 Results: Results obtained identifies 18(81.8%) of the ESBL isolates to be harbouring plasmids of different molecular weights (2-21.5kbp). The total protein profile of the plasmid-encoded ESBL and the non-plasmid encoded ESBL reveals differential protein expression patterns except for two non- plasmid encoded ESBL isolates that have similar patterns as that of plasmid-encoded ESBL organisms (isolates 2 and 3).
 Conclusion: There is a need to monitor the primary sources of infection in the epidemiological distribution of resistance. Improving environmental sanitation through proper disinfection of carrier insects may be a means to curtailing such spread.

Prevalence of extended-spectrum beta-lactamase-producing Enterobacteriaceae among clinical isolates in Turaif general hospital, northern borders- Saudi Arabia.

Enterobacteriaceae that produce extended-spectrum beta-lactamase (ESBL) are quickly spreading, posing a threat to world healthcare. 138 gram-negative bacteria were collected from different samples (stool, urine, wound, blood, tracheal aspirate, catheter tip, vaginal swab, sputum, and tracheal aspirate) from hospitalized patients. Samples were subcultured and identified in accordance with their biochemical reactions and culture characteristics. Against all the isolated Enterobacteriaceae, an antimicrobial susceptibility test was performed. VITEK®2 system, phenotypic confirmation, and Double-Disk Synergy Test (DDST) had been utilized to identify the ESBLs. Of the 138 samples studied, the prevalence of ESBL-producing infections among the clinical samples of the present study was 26.8 % (n = 37). E. coli was the commonest ESΒL producer at 51.4% (n = 19) followed by K. pneumoniae at 27% (n = 10). The potential risk factors for the ESBL development that produces bacteria were as follows, patients with the presence of indwelling devices, previous history of hospital admission, and usage of antibiotics. ESBL is statistically (p ≤ 0.05) higher among the patients with indwelling devices, ICU admission, who had a previous hospital admission in the last 6 months as well as who was given antibiotics (quinolones and/or cephalosporins) in the last 6 months. One hundred thirty-two (95.7%) of ESBL isolates were resistant to amoxicillin, while the lowest resistance was for fosfomycin (15.2%). ESBL-producing Enterobacteriaceae are highly prevalent in Turaif General Hospital setting with some potential risk factors. A strict policy to be made available on the usage of antimicrobials in hospitals and clinics should be established.

Risk factors of late onset sepsis caused by extended spectrum beta-lactamase (ESBL) - producing bacteria in preterm infants

Background High incidence of late-onset sepsis (LOS) in preterm infants contributes to neonatal morbidity. Therapeutic outcomes of LOS have deteriorated as a result of increased antibiotic resistance problems, mainly from ESBL isolates. Controlling risk factors is important in reducing morbidity and mortality as well as providing guidance for antibiotic selection.
 Objectives To determine the risk factors of LOS due to ESBL-producing bacteria in preterm infants.
 Methods This is a retrospective study. The inclusion criteria was neonates diagnosed with late-onset neonatal sepsis by clinical signs and a positive blood culture. The blood culture result and characteristics patients as secondary data were extracted from medical records within the hospital facilities and the institutional database of the Neonatology Department of Universitas Brawijaya (January 2019 to March 2021). Statistical analysis was done to compare characteristics of the patients in the ESBL positive group to those in the ESBL negative group to assess the potential risk factors.
 Results Among 124 preterm infants with LOS, 62 of them were ESBL-positive case subjects and the other 62 were non-ESBL-producing control subjects. Gram-negative bacteria were the most common pathogens identified, with 96% (n=59) of them being the ESBL-producing strain, predominated by Klebsiella pneumoniae (n=56). Factors significantly correlated with the occurrence of LOS-ESBL included prior history of invasive procedures (OR 3.13; 95%CI 1.45 to 6.73; P=0.00), central access insertion (OR 9.54; 95%CI 3.7 to 24.2; P=0.00), and parenteral nutrition (OR 6.03; 95%CI 2.77 to 13.16; P=0.00). Central access insertion had the strongest influence (Exp(B) 6.98; P= 0.00).
 Conclusion Prior invasive procedures, central access insertion, and parenteral nutrition had significant correlations with the occurrence of LOS-ESBL in preterm infants. Central access insertion is a predictive factor for LOS-ESBL.

Incidence and molecular characterization of extended-spectrum beta-lactamase (ESBL)-producing Salmonella enterica and Escherichia coli of avifauna origin in Pakistan.

Members of Enterobacteriaceae are known to produce extended-spectrum beta-lactamases (ESBL) which hydrolyze the beta-lactam group of antibiotics. The existence of ESBL-producing Salmonella enterica (S. enterica) and Escherichia coli (E. coli) harbored by urban avifauna was investigated in this study. Dropping samples (n= 180) were collected from six different bird species in the district Jhang, Punjab province, Pakistan. Isolation and identification of ESBL isolates were made by using cefotaxime- (4 mg/L) supplemented MacConkey agar and double disc synergy test (DDST). Polymerase chain reaction (PCR) was performed for the detection of four different ESBL genes including blaCTX-M, blaTEM, blaSHV and blaOXA. A total of 42.69% isolates were confirmed as ESBL via DDST including 30.64% S. enterica and 49.54% E. coli. The incidence of ESBL S. enterica and ESBL E. coli was found highest in egret (Ardea alba) and pigeon (Columba livia) as 64.28% and 78.95%, respectively. The blaCTX-M gene was detected in 57.89% and 64.81% of isolates of S. enterica and E. coli, respectively. Among other genes in S. enterica and E. coli, blaTEM (21.05%, 20.4%); blaSHV (15.78%, 9.26%), and blaOXA (5.26%, 5.56%) were detected, respectively. All of the tested isolates were found resistant to at least one of the thirteen antimicrobial agents except meropenem. To the best of our knowledge, this is the first study reporting the incidence and genetic diversity of ESBL bacteria associated with urban avifauna in Pakistan. The urban avifauna can serve as a potential subject of bio-surveillance to monitor the emergence of antimicrobial-resistant bacteria.

Biofilm Formation Status in ESBL-Producing Bacteria Recovered from Clinical Specimens of Patients: A Systematic Review and Meta-Analysis.

Recently, the emergence and spread of extended-spectrum beta-lactamase (ESBL) bacteria have become a global health concern. In addition, the ability to form biofilm due to less impermeability to antibiotics and the horizontal transformation (conjugation) of genes involved in antibiotic resistance have exacerbated the concerns. With a comprehensive meta-analysis, this study evaluated the potential relationship between ESBL and biofilm formation. A literature search was performed using global databases, such as PubMed and Scopus, up to November 2021. We retrieved all relevant documents and selected eligible articles based on inclusion criteria. Finally, the potential association between the biofilm formation capacity and resistance of ESBL-producing bacteria was measured with an odds ratio and a 95% confidence interval. In the present study, 17 articles, including 2,069 Gram-negative isolates, were considered as eligible. The prevalence of biofilm formation in all clinical isolates of ESBL and non-ESBL pathogens was 72.4% (95% CI: 60.7-81.6) and 40.5% (95% CI: 30.2-51.8), respectively. Our results showed a positive relationship between the ability for biofilm formation and conferring antibiotic resistance in ESBL-producing bacteria (OR: 3.35; 95% CI: 1.67-6.74; p-value: 0.001). In general, we showed the rate of biofilm formation to be significantly higher in ESBLproducing strains. Given the current results, the updated therapeutic guidelines should consider the role of biofilm production for optimal therapy, treatment course, and clinical outcomes rather than the recommendation of antimicrobial agents by focusing on the results of the antibiotic susceptibility test.

Molecular Characterization and Antimicrobial Resistance Profiling of Extended Spectrum Beta-lactamase (ESBL) Producing Escherichia coli in Bovines from J and K, India

Background: Higher prevalence of ESBL producers is alarming in dairy sector. So limiting antimicrobial use may curtail the selection and persistence of predominant ESBL and conjugative plasmids among strains. The study was aimed to determine the occurrence of extended-spectrum beta-lactamase (ESBL) producing E. coli as well as their genetic diversity, antimicrobial resistance and integrons in bovines. Methods: A total of 180 faecal samples were screened for the presumptive ESBL producing E. coli isolates. All ESBL producing isolates were subjected to the screening test and were confirmed as ESBL producers by double disc. Further, these ESBL producers were tested for the presence of bla genes and those found positive were tested for multidrug resistance (MDR) by disk diffsion. MDR positive isolates were further tested for the presence of intI 1, intI 2 and intI 3 genes. Result: A total of 360 presumptive ESBL producing E. coli isolates were obtained. Out of which, 154 (42.77%) isolates were found to be resistant and were confirmed as ESBL producers. Of 154 ESBL isolates, 120 (77.92%) isolates carried the gene/s screened for blaTEM, blaCTX-M and blaSHV genes are were declared as multi drug resistant. Out of 120 MDR isolates, 59 tested positive for the integron gene.

Phylogenetic groups, serogroups and virulence factors of uropathogenic Escherichia coli isolated from patients with urinary tract infection in Baghdad, Iraq.

Uropathogenic Escherichia coli (UPEC) is divided into different phylogenetic groups that differ in their antibiotic resistance patterns, serogroups and pathogenicity. This study aimed to investigate the prevalence of phylogenetic groups of UPEC isolates and their relationship with serogroups and virulence factors in patients with UTIs. Of the 412 urine samples tested a total of 150 UPEC were isolated and confirmed with PCR using 16S rRNA gene. Antibiotic resistance of the isolates was tested using disk diffusion method and the isolates were divided into phylogenetic groups by the quadruplex PCR method. The prevalence of serogroups and virulence genes were investigated using multiplex PCR. 87 (58%) of the isolates belonged to phylogroup B2. Virulence genes fimH (95.3%), aer (49.3%) and serogroups O8 (22.3%), O25 (21.5%) showed the highest prevalence. The lowest drug resistance was observed against imipenem (4.6%) and meropenem (3.3%). The prevalence of multidrug-resistant and extended-spectrum beta-lactamases isolates were 60% and 61.3%, respectively. We also found a significant relationship between phylogenetic groups, serogroups and virulence factors among our isolates. The high abundance of phylogenetic group B2, serogroups O8 and O25, and virulence genes fimH and aer indicate their importance in the pathogenesis of UPEC in this country.

Antibiotic susceptibility profiles and detection of genes mediating extended-spectrum β-lactamase (Esbl) production in escherichia coli isolates from National Hospital, Abuja.

Extended-spectrum beta-lactamases (ESBL) are the most widespread of the new beta-lactamases and are a significant threat to patient care in the hospital and the community. Aim: The study assessed the prevalence, antibiotic susceptibility profile, and major ESBL encoding genes among Escherichia coli isolated from clinical specimens of patients in the National Hospital, Abuja (NHA). E. coli isolated from diverse clinical specimens obtained from clinically proven cases of infection managed at the NHA were included in the study. The antimicrobial susceptibility was performed by the Kirby-Bauer method and E-test was used to confirm the ESBL phenotype. Multiplex polymerase chain reaction (PCR) was used to detect the genes mediating ESBL production. Meropenem, fosfomycin, and tigecycline demonstrated excellent activities against all isolates: of the 400 isolates, 392 (98%), 386 (96.5%), and 362 (90.5%) were susceptible, respectively. Similarly, 358 (89.5%) were susceptible to amikacin, 323 (80.3%) nitrofurantoin, 281 (70.3%) ceftazidime, and 279 (69.8%) cefotaxime. A total of 271 (67.8%), 219 (54.8%), and 208 (52.0%) were resistant to amoxicillin-clavulanate, ciprofloxacillin, and gentamicin, respectively. However, all the isolates were resistant to ampicillin. There was a significantly higher proportion of multidrug resistance among ESBL-producing isolates compared to non-ESBL-producing isolates (P = 0.0001). Of the 121 phenotypically detected ESBL isolates, 119 (98.3%) harbored genes mediating the production of Cefotaximase- Munich (CTX-M), Temoniera (TEM) or Sulfhydryl Variable (SHV) enzymes. The prevalence of ESBLs among E. coli was relatively high, at 30.2%. About 81% of all blood isolates were ESBL-producers. blaCTX-M is the predominant type of ESBL gene among E. coli. A high proportion of the ESBL-producing isolates expressed a combination of two or three genes together.

A Three-Year Look at the Phylogenetic Profile, Antimicrobial Resistance, and Associated Virulence Genes of Uropathogenic Escherichia coli.

Uropathogenic Escherichia coli is the most common cause of urinary tract infections, resulting in about 150 million reported annual cases. With multidrug resistance on the rise and the need for global and region surveillance, this investigation looks at the UPEC isolates collected for a 3-year period, with a view of ascertaining their antimicrobial susceptibility patterns and associated virulence determinants. The identification of bacteria isolates, antimicrobial susceptibility, and extended-spectrum beta-lactamases (ESBLs) production was determined with a Vitek 2 Compact Automated System (BioMerieux, Marcy L’Etoile, France). ESBLs were confirmed by the combined disc test (CDT) and basic biochemical test. The isolates were distributed into A (11%), B1 (6%), B2 (62.4%), and D (20.6%). Resistance to the penicillin group was high, between 88% and 100%. Additionally, resistance was high to cephalosporins (100%) in 2017 and 2018. The isolates were all sensitive to tigecycline, while resistance against imipenem and meropenem was low, at 4–12% in 2017 and 2018 and 0% in 2019. The results also showed that ESBL isolates were seen in 2017 and 2018. They were confirmed positive to CTX/CLA (88.5%) and CAZ/CLA (85%). By 2019, the number of resistant isolates reduced, showing only 4% ESBL isolates. Two virulence genes, fimH (46%) and papE/F (15%), were detected among the isolates by PCR. In conclusion, this study found that phylogroups B2 and D carried the most virulence genes as well as MDR and ESBL characteristics, suggesting the UPEC strains to be extraintestinal pathogens responsible for UTIs.

Increased rates of extended-spectrum beta-lactamase isolates in patients hospitalized with culture-positive urinary Enterobacterales in the United States: 2011 – 2020

Antimicrobial resistance in Enterobacterales has made empiric therapy for hospitalized patients with urinary tract infections (UTIs) more challenging. We analyzed the antibiotic susceptibility of nonduplicate Enterobacterales isolates from urine cultures tested at US hospitals in the BD Insights Research Database (2011–2020). Multivariable generalized estimating equation models were used to assess resistance trends over time. A total of 322 US hospitals provided data on 876,507 urinary Enterobacterales isolates (62.4% Escherichia coli). Enterobacterales antibiotic resistance rates were 64.6%, 29.3%, 27.6%, and 26.3% for beta-lactams, fluoroquinolones, nitrofurantoin, and trimethoprim/sulfamethoxazole, respectively, and 12.4% had an extended-spectrum beta-lactamase (ESBL) phenotype. In multivariable models, rates of ESBL isolates and isolates resistant to ≥3 drug classes increased significantly between 2011 and 2020, while other categories of resistance generally decreased. We conclude that antimicrobial resistance is common in urinary Enterobacterales isolates. Management of UTIs should be guided by urine culture data and may benefit from new therapies.

Prevalence of Extended Spectrum Beta Lactamase producing Escherichia coli and Klebsiella spp. from urinary specimen in a tertiary care hospital

Escherichia coli and Klebsiella spp. are two important uropathogens showing resistance to beta-lactams by producing extended spectrum beta lactamase (ESBL), leading to difficulty in treating infections with these bacteria. Majority of the ESBLs are of TEM, CTXM and SHV types reported from Nepal. This study was conducted in Clinical Microbiology Department of Nepal Medical College Teaching Hospital from November 2021 to mid-January 2022 to determine prevalence of ESBL and antimicrobial susceptibility pattern among two common uropathogens. A total of 402 E. coli and Klebsiella spp. isolated from urine samples were included in the study. ESBL producing strains were identified by combination disc method and antibiotic susceptibility testing was done by Kirby Bauer’s disc diffusion method. PCR was done to detect the TEM and CTXM genes. Prevalence of ESBL among E. coli and Klebsiella spp. was found to be 14.9% (60/402). Among the E. coli, 16.1% (58/360) and only 4.8% (2/42) of Klebsiella spp. were ESBL producers. Antibiogram showed, 56.7% ESBL producers were susceptible to ciprofloxacin, 46.7% to co-trimoxazole, 96.7% to nitrofurantoin. However, all isolates were susceptible to amikacin, piperacillin-tazobactam, and carbapenems (both meropenem and imipenem). Among total 30 ESBL isolates selected by systematic random sampling, 3 isolates (all three E. coli) showed presence of TEM and 3 other isolates (two E. coli and one Klebsiella spp.) showed CTXM genes.