Coli Isolates Research Articles

Molecular Detection of Virulence-Associated Markers in Campylobacter coli and Campylobacter jejuni Isolates From Water, Cattle, and Chicken Faecal Samples From Kajiado County, Kenya.

Campylobacter is a zoonotic foodborne pathogen that is often linked with gastroenteritis and other extraintestinal infections in humans. This study is aimed at determining the genetic determinants of virulence-encoding genes responsible for flagellin motility protein A (flaA), Campylobacter adhesion to fibronectin F (cadF), Campylobacter invasion antigen B (ciaB) and cytolethal distending toxin (cdt) A (cdtA) in Campylobacter species. A total of 29 Campylobacter coli isolates (16 from cattle, 9 from chicken, and 4 from water samples) and 74 Campylobacter jejuni isolates (38 from cattle, 30 from chicken, and 6 from water samples) described in an earlier study in Kajiado County, Kenya, were examined for the occurrence of virulence-associated genes using polymerase chain reaction (PCR) and amplicon sequencing. The correlations among virulence genes were analyzed using Pearson's correlation coefficient (R) method. Among the 103 Campylobacter strains screened, 89 were found to harbour a single or multiple virulence gene(s), giving an overall prevalence of 86.4%. C. jejuni strains had the highest prevalence of multivirulence at 64.9% (48/74), compared to C. coli (58.6%, 17/29). The ciaB and flaA genes were the most common virulence genes detected in C. jejuni (81.1% [60/74] and 62.2% [46/74], respectively) and in C. coli (each at 62.1%; 18/29). Campylobacter isolates from chicken harboured the most virulence-encoding genes. C. jejuni strains from chicken and cattle harboured the highest proportions of the cdtA and ciaB genes, respectively. All the C. coli strains from water samples harboured the cadF and flaA genes. The results obtained further revealed a significant positive correlation between cadF and flaA (R = 0.733). C. jejuni and C. coli strains from cattle, chicken, and water harbour virulence markers responsible for motility/colonization, invasion, adherence, and toxin production, evoking their important role in campylobacteriosis development among humans and livestock. The identification of cattle, chicken, and water samples as reservoirs of virulent Campylobacter spp. highlights the possible risk to human health. These data on some virulence genes of Campylobacter will assist food safety and public health officials in formulating policy statements.

Profiling of antimicrobial resistance (AMR) and multi drug resistance (MDR) of <em>Escherichia coli</em> isolates from livestock, wild animals and an ornamental fish farm in Polgahawela, Sri Lanka

The pattern of antimicrobial resistance (AMR) and the prevalence of multi drug resistance (MDR) of Escherichia coli (E. coli) isolates from livestock, wild animals and an ornamental fish farm were investigated in a selected area in Polgahawela, Sri Lanka. Sampling area of one square Km containing an ornamental fish farm, a broiler farm, cattle, goat, and backyard poultry which are located separately within the sampling area but not in an integrated farming system with its associated wildlife was mapped using GPS. Faecal samples were examined from 89 wild animals (terrestrial and avian species), 28 livestock animals (including poultry) as well as 40 samples from an ornamental fish farm (fish faecal material, pond water and pond sediment). E. coli was isolated using standard methods and AMR profiles were investigated for 12 antimicrobials using Kirby-Bauer disc diffusion method. Sixty nine percent (62/89) of wild animals and 86% (24/28) of livestock samples were positive for E. coli whereas the isolation rate was 17.5% (7/40) in the samples from the ornamental fish farm. A maximum of two randomly selected E. coli isolates from each sample, making a total of 164 isolates were tested for AMR of which 114, 43 and 7 were from wild animals, livestock and the ornamental fish farm, respectively. The E. coli isolates tested against 12 antimicrobials reflected that the proportions of E. coli isolates resistant to at least one antimicrobial were 30%, 63% and 71% in wild animals, livestock and the ornamental fish farm, respectively. Multidrug-resistant E. coli isolates were detected in wild animals, livestock and the samples from ornamental fish farm. Overall, according to the statistical analysis, E. coli isolated from livestock and the ornamental fish farm reflected a significantly higher chance of carrying AMR compared with E. coli from wild animals.

BlaCTX-M Mediated Extended Spectrum Beta-lactamase Production in Escherichia coli from Urine Samples

Objective: To assess the antimicrobial pattern of Escherichia coli isolates from urine samples, determine the proportion of multidrug resistant (MDR) and extended spectrum beta-lactamase (ESBL) and blaCTX-M gene of E. coli from the urine samples. Methods: A total of 2452 urine samples were collected and processed from April to October 2023 in clinically suspected urinary tract infection (UTI) patients attending B&B Hospital Pvt. Ltd to identify possible bacteria using standard microbiological procedures. E coli isolates were further subjected to antimicrobial susceptibility testing using Kirby Bauer disk diffusion method. Combination disk method was used to identify the ESBL phenotypes. Molecular detection of blaCTX-M gene was performed using conventional polymerase chain reaction. Results: A total of 338 (13.7%) samples showed significant bacterial growth. Maximum bacterial growth was found in outpatients (70.4%) and in patients above 60 years of age. Gram negative bacteria (87.8%) were predominant and E. coli (54.4%) was the most frequent. Nitrofurantoin, Aminoglycosides, Carbapenems and Polymixins were effective antibiotics. Among 184 E. coli isolates, 54 (29%) were MDR, 30 (16.3%) were ESBL producer. blaCTX-M gene was detected in 90% of the phenotypically confirmed ESBL producers. Conclusion: E. coli is the major cause of UTI and blaCTX-M is a prime contributor of ESBL production. High rate of resistance to third-generation Cephalosporins, Flouroquinolones and Cotrimoxazole make these antibiotics unsuitable for the treatment of UTI.

A Prospective Study to Identify Asymptomatic Bacteriuria: The Causative Organisms, Antibiotic Sensitivity and its Safety in Pregnant Females

Introduction: The common procedure of urinary catheterization during childbirth may introduce bacteria that can cause urinary tract infections. Immune function is relatively compromised during pregnancy. The current study was aimed to identify antenatal patients with bacteriuria, the causative organism and its antibiotic sensitivity in patients with bacteriuria. Materials and Methods: The current prospective crosssectional study involved antenatal patients who were being admitted to the delivery room. A clean catch midstream urine sample was collected in a sterile container of which 5 to 10 mL of the sample was obtained in a separate sterile vial (container 1) for screening tests for leucocyte esterase and nitrite, and the remaining sample (container 2) was taken for routine microscopy, culture, and sensitivity. The reports were collected and tabulated, and bacteriuria was confirmed and statistically analyzed. Results: A total of 200 patients were evaluated, and 40 of them had asymptomatic bacteriuria. In this study positive leukocyte esterase test and nitrite test results were considerably higher in the cases than the controls (p < 0.05) at 10.0 and 47.5%, respectively. Escherichia coli was found to be the most prevalent bacteria in this study, followed by, Pseudomonas, Enterococcus faecalis, Candida albicans, and coagulasenegative Streptococcus and Klebsiella. Most of the E. coli isolates (100.0%) were sensitive to amikacin, a category D drug. The majority of Pseudomonas isolates (100.0%) were sensitive to colistin, a category C drug. Amongst category A drugs, Pseudomonas was sensitive to – cefoperazone-sulbactam, meropenem and piperacillin-tazobactam. Conclusion: It is a standard practice to send urine routine and microscopy test in antenatal patients. As most patients are asymptomatic, investigations are required for early diagnosis of significant bacteria like leucocyte esterase, nitrite and culture and sensitivity. This will help us to institute appropriate treatment and improve maternal and fetal outcomes.

Characterization of Campylobacter jejuni and Campylobacter coli isolates from chicken offal in Metro Manila, Philippines: Insights from virulence gene prevalence and multilocus sequence typing analysis.

Campylobacteriosis is a human infection primarily caused by Campylobacter jejuni and Campylobacter coli. Consumption of contaminated chicken and poultry products is the main mode of transmission. These bacteria possess virulence factors, including adhesins and toxins, which contribute to their pathogenesis. Moreover, their large genomes undergo frequent genetic recombination, resulting in a high degree of genetic diversity. However, limited information is available regarding the virulence and genotypic diversity profiles of these microorganisms in the Philippines. The objective of this study was to address this knowledge gap by characterizing Campylobacter isolates obtained from chicken offal sold in wet markets in Metro Manila, Philippines. Multilocus Sequence Typing (MLST) analysis was performed to determine the sequence types, resulting in the identification of 13 unique sequence types, including nine previously unreported ones, and three clonal complexes. Notably, the widespread sequence type ST-305 was found in samples from different markets. Furthermore, six isolates deposited in the Campylobacter PubMLST database were identified as C. coli based on allele profiles. Profiling using 10 selected virulence genes revealed that more than half of the isolates carried these genes. The most prevalent virulence gene was cadF (100%), followed by flaA (95%), racR, cdtA, cdtB, and cdtC (85%). The genes dnaJ and ceuE were also present in 75% of the isolates. Despite the limited sample size, the findings of this study reveal a significant level of genotypic diversity among the Campylobacter isolates. This diversity has important implications for source attribution studies and the identification of strains involved in campylobacteriosis outbreaks. Furthermore, the investigation of virulence factors associated with colonization and invasion of the avian gut can provide insights for the development of practical applications in Campylobacter control strategies. Understanding and addressing these factors are crucial steps toward mitigating the risk of Campylobacter infections and enhancing public health efforts.

ASSOCIATION OF TLR 5 AND ESCHERICHIA COLI FLIC POLYMORPHISMS WITH RECURRENT URINARY TRACT INFECTIONS IN WOMEN

This study was aimed to investigate the role of some Toll-like receptor 5 (TLR5) and E. coli fliC gene polymorphisms with increased risk to recurrent urinary tract infections (rUTI). From 180 specimens (blood and urine) were collected from women of different age, 60 of them serve as control while 120 had rUTI symptoms. After culturing of urine specimens, 43 (35.8%) were identified as E. coli isolates. Four SNPs were identified when amplified and sequenced of TLR5 include rs5744168, rs775385356, rs2072493 and rs5744174. Twelve flagellar antigen serotypes were obtained from fliC sequence of 28 isolates. By using Expasy and Clustal Omega programs, the FliC proteins of H-serotypes arranged in different lengths ranging between 324-634 residues. The N-terminal and C-terminal were conserved region, in contrast, the central region was variable poorly preserved. The results also showed list of conserved amino acids in both FliC termini included L89, Q90,R91, L95, Q98, N101 and E115 in N- terminus in all studied serotypes. Further, the N277 residue that reported its central role for TLR5 interaction were found in some serotypes. Indeed, it was no distinct relation between genotypes or allele frequencies with fliC polymorphisms found in E. coli isolated from rUTI.

Clinical and microbiological characteristics of urinary tract infections in the first year after renal transplantation

Urinary tract infections (UTI) are the most frequent infection in kidney transplant (KT) patients. The main objective was to determine the clinical and microbiological characteristics of UTIs that occur during the first year after KT. Retrospective cohort study that included patients over 18 years of age who received KT between 2009-2020 in two hospitals in Córdoba city, Argentina. Patients were followed up during the first year after the transplant by recording them in the medical records, and those who presented at least one episode of UTI were analyzed. During the study period, 568 KTs were performed, of which 207 (36.4%) had at least one episode of UTI. In total, there were 419 UTI episodes, 6 (1.4%) polymicrobial UTI episodes, so a total of 426 microorganisms were identified in the urine cultures. Of the total number of episodes, 206 (49.2%) occurred between 31 and 180 days post-transplant. The main etiological agent was E. coli with 225 isolates (52.8%) followed by Klebsiella sp. with 94 (22.1%). 52.1% of all episodes were caused by multi-resistant microorganisms (MRM). Among the E. coli isolates, 94 (41.8%) were MRM. In the multivariate analysis, the risk factors associated with UTI due to MRM were a history of recurrent UTI (Odds ratio 2.43; 95% CI: 1.37-4.30) and induction with basiliximab (Odds ratio 1.53; 95% CI: 1.029-2.29). UTIs occurred in more than a third of kidney transplant patients, being slightly more than half caused by MOR.

Synthesis, Antimicrobial and Computational Studies of New Branched Azaphenothiazinones Derivatives

In a continued search for new medicinally active nonlinear phenothiazines, novel angular chloroazaphenothiazinone derivatives have been synthesized via transition metal-catalyzed cross-coupling reactions. The structural elucidation of the synthesized compounds was established by a combined spectroscopic and elemental analysis. The synthesized compounds were tested for their antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, Enterococusfaecalis, Escherichia coli, Candida albican,and Aspergillus niger isolates by the convectional agar-well dilution method and compounds 5c and 8cdisclosed excellent in vitro activity against some of the tested microorganisms. In silico,the study showed that the synthesized compounds possessed promising physichemical properties and fit well in the active site of a Biotin-Protein Ligase (BPL) forming essential hydrogen bonding and hydrophobic interactions.

Isolation and Characterization of Cefotaxime and Ciprofloxacin Co-Resistant Escherichia coli in Retail Chicken Carcasses

Transmission of antimicrobial-resistant bacteria to humans through the food chain is of great importance for public health. In this study, it was aimed to isolate and characterize the cefotaxime and ciprofloxacin-resistant Escherichia coli in retail chicken meat samples sold in Hatay. The isolates were subjected to phylogenetic group typing and antimicrobial susceptibility testing. The genetic relatedness of the isolates was determined using Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) technique. The isolates were also screened for the presence of both antimicrobial and plasmid-mediated quinolone resistance (PMQR) genes by PCR. Cefotaxime and ciprofloxacin co-resistant E. coli isolates with diverse genetic origins were recovered in 42.3% (22/52) of retail chicken carcasses. The E. coli isolates belonged to the phylogenetic group D2 were dominant (40.9%, 9/22), followed by B1 (27.3%, 6/22), B23 (18.2%, 4/22), and A1 (13.6%, 3/22), respectively. Based on dendrogram analysis, the ERIC-PCR method differentiated the isolates into 10 clusters (I-X). The multidrug resistance (MDR) was observed in 81.8% (18/22) of the isolates. PMQR determinants were not identified in any isolates tested. Molecular analysis revealed one or more β-lactamase-encoding genes in all isolates as a single or in combination: blaCTX-M-blaTEM (n=5), blaCMY-2 (n=5), blaCTX-M (n=5), blaCMY-2-blaSHV (n=3), blaCMY-2-blaTEM (n=3), and blaCTX-M-blaCMY-2 (n=1). This study highlights that retail chicken meat is an important reservoir of cefotaxime and ciprofloxacin co-resistant E. coli isolates. It is necessary to evaluate their contribution to the community and hospital infections.

UspA gene-based phylogenetic analysis and antigenic epitope prediction for Escherichia coli strains of avian origin.

Pathogenic Escherichia coli (E. coli) is responsible for various local and systemic infections in animal and human populations. Conventional methods for the detection and identification of E. coli are time-consuming and less reliable for atypical strains. The uspA gene has been widely used as a target for the detection of E. coli. The present study was aimed at phylogenetic analysis of the uspA gene sequences to determine the evolutionary relationships between the strains and other members of the Enterobacteriaceae family. In addition, the unique differences in the sequences of the current study with Salmonella and Shigella species were tested using Tajima's molecular clock test. Antigenic epitope prediction was performed to locate the B-cell epitope region of the UspA protein. Two E. coli isolates of avian origin and strains from the National Center for Biotechnology Information (NCBI) database were used for prediction. The Immune Epitope Database (IEDB) server, Bepitope, ABCpred, SVMTrip, and ElliPro server were used to identify B-cell epitopes. The 3D structure was predicted using SWISS-MODEL. Phylogenetic analysis of the isolates from the current study revealed that both OM837340 and OM837341 sequences from the current study had maximum nucleotide homology (nt) of 99.87%-100% with E. coli isolates and minimum nt homology of 84.08% with Salmonella enteritidis and S. Hissar. The isolates in the current study had a homology of 98.87%, while the homology with Shigella species was 99.25%. Seven silent mutations were observed in the coding region of the UspA protein of ECO9LTBW (current study). Modeling of the UspA protein revealed a maximum homology of 67.86% with the Protein Data Bank in Europe (PDBe), also validated by the Ramachandran plot. No significant differences were found in the coding regions of uspA of Salmonella, Shigella, and E. coli with Tajima's test. For the E. coli isolates, a total of 24 linear B-cell and seven discontinuous epitopes were predicted using in-silico analysis. When the results of the predicted peptides were compared, two peptides, namely ARPYNA and YSDLYTGLIDVNLGDMQKRISEE, were found suitable candidates. In conclusion, the uspA gene appears to be conserved among E. coli isolates and can be used for molecular detection.

Within-host genetic diversity of extended-spectrum beta-lactamase-producing Enterobacterales in long-term colonized patients

Despite recognition of the immediate impact of infections caused by extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales (ESBL-PE) on human health, essential aspects of their molecular epidemiology remain under-investigated. This includes knowledge on the potential of a particular strain to persist in a host, mutational events during colonization, and the genetic diversity in individual patients over time. To investigate long-term genetic diversity of colonizing and infecting ESBL-Klebsiella pneumoniae species complex and ESBL-Escherichia coli in individual patients over time, we performed a ten-year longitudinal retrospective study and extracted clinical and microbiological data from electronic health records. In this investigation, 76 ESBL-K. pneumoniae species complex and 284 ESBL-E. coli isolates were recovered from 19 and 61 patients. Strain persistence was detected in all patients colonized with ESBL-K. pneumoniae species complex, and 83.6% of patients colonized with ESBL-E. coli. We frequently observed isolates of the same strain recovered from different body sites associated with either colonization or infection. Antimicrobial resistance genes, plasmid replicons, and whole ESBL-plasmids were shared between isolates regardless of chromosomal relatedness. Our study suggests that patients colonized with ESBL-producers may act as durable reservoirs for ongoing transmission of ESBLs, and that they are at prolonged risk of recurrent infection with colonizing strains.

Phylogenetic monitoring of Escherichia coli in medical wastewater/Baghdad City

Clean drinking water access is a main factor that supports general health worldwide, where important investment is made to maintain water quality. The gene sequence of 16S rRNA was applied to study bacterial phylogenesis and classification. This studyaimed to isolate new bacterial strains from wastewater environments. Thisstudy collected water samples from differentwastewater sites for Baghdad Medical City’s Hospital. The results of Vitek 2Compact were with a probability of99% forEscherichia coli isolates .The bacterial isolates were identified using Polymerase chain reaction (PCR) based on the universaldiagnostic gene 16SrRNA, and the PCR product was obtained with a molecular weight of 1250bp. The PCR productsequencingshowed 166 isolates of E. coli different from the isolates registered in the NCBI database of E. coli after alignment between them.The present study reported 16 Iraqi isolates in NCBI with an accession number: OM032663.1; OM 032664.1; OM294659.2;ON724178.1; ON724264.1; ON724331.1; ON725076.1; ON725091.1; ON725139.1; ON725141.1; ON739200.1; ON739201.1;ON739207.1; ON739208.1; ON739209.1; ON739211.1. The identity was then observed(96-100%) with strains in Gene Bank.There were many different sequence position substitutions (transition, deletion, transversionfor the registered local isolates whenconducting multiple alignments and comparing themwith the isolates in the Gene Bank. According to the phylogenetic treebuilding by joining-the neighbor method, the local Iraqi isolates clustered into sub-descents. Moreover, local isolates of E.coliappeared more convergent to Saudi Arabia isolates (KY780347.1). This may be due to the geographical proximity of the two countries to the same Arab Gulf region .The present study identified 16 isolates of E. coli from the Iraqi aquatic environment (medical wastewater). The novelty of this study was represented to monitoring the evolution of E. coli in medical wastewater.

Tigecycline Sensitivity Reduction in Escherichia coli Due to Widely Distributed tet(A) Variants.

Despite scattered studies that have reported mutations in the tet(A) gene potentially linked to tigecycline resistance in clinical pathogens, the detailed function and epidemiology of these tet(A) variants remains limited. In this study, we analyzed 64 Escherichia coli isolates derived from MacConkey plates supplemented with tigecycline (2 μg/mL) and identified five distinct tet(A) variants that account for reduced sensitivity to tigecycline. In contrast to varied tigecycline MICs (0.25 to 16 μg/mL) of the 64 tet(A)-variant-positive E. coli isolates, gene function analysis confirmed that the five tet(A) variants exhibited a similar capacity to reduce tigecycline sensitivity in DH5α carrying pUC19. Among the observed seven non-synonymous mutations, the V55M mutation was unequivocally validated for its positive role in conferring tigecycline resistance. Interestingly, the variability in tigecycline MICs among the E. coli strains did not correlate with tet(A) gene expression. Instead, a statistically significant reduction in intracellular tigecycline concentrations was noted in strains displaying higher MICs. Genomic analysis of 30 representative E. coli isolates revealed that tet(A) variants predominantly resided on plasmids (n = 14) and circular intermediates (n = 13). Within China, analysis of a well-characterized E. coli collection isolated from pigs and chickens in 2018 revealed the presence of eight tet(A) variants in 103 (4.2%, 95% CI: 3.4-5.0%) isolates across 13 out of 17 tested Chinese provinces or municipalities. Globally, BLASTN analysis identified 21 tet(A) variants in approximately 20.19% (49,423/244,764) of E. coli genomes in the Pathogen Detection database. These mutant tet(A) genes have been widely disseminated among E. coli isolates from humans, food animals, and the environment sectors, exhibiting a growing trend in tet(A) variants over five decades. Our findings underscore the urgency of addressing tigecycline resistance and the underestimated role of tet(A) mutations in this context.

Prevalence and antimicrobial resistance profiles of extended-spectrum beta-lactamase-producing Escherichia coli in East Tennessee dairy farms.

The extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae, such as Escherichia coli, are emerging as a serious threat to global health due to their rapid spread and their multidrug-resistant (MDR) phenotypes. However, limited information is available regarding the prevalence and antimicrobial resistance (AMR) profile of ESBL-E. coli in the United States dairy farms. This study aimed to determine the prevalence and AMR pattern of ESBL-E. coli in East Tennessee dairy cattle farms. Rectal fecal samples from dairy cattle (n = 508) and manure (n = 30), water (n = 19), and feed samples (n = 15) were collected from 14 farms. The presumptive E. coli was isolated on CHROMagar™ ESBL and confirmed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Antimicrobial susceptibility testing was performed on the ESBL-E. coli isolates. From 572 fecal and farm environmental samples, a total of 233 (41%, n = 572) ESBL-E. coli were identified. The prevalence of fecal ESBL-E. coli was 47.5% (95% CI: 46.2-49.2). The within-farm prevalence of ESBL-E. coli ranged from 8 to 100%. Recent treatment history with third-generation cephalosporins (3GC), cow parity ≥3, and calves were the independent risk factors associated (P < 0.05) with fecal carriage of ESBL-E. coli. Overall, 99.6% (n = 231) ESBL-E. coli tested were phenotypically resistant to at least one of the 14 antimicrobial agents tested. The most common AMR phenotypes were against beta-lactam antibiotics, ampicillin (99.1%; n = 231 isolates), and ceftriaxone (98.7%, n = 231). Most ESBL-E. coli isolates (94.4%) were MDR (resistance to ≥3 antimicrobial classes), of which 42.6% showed co-resistance to at least six classes of antimicrobials. ESBL-E. coli isolates with concurrent resistance to ceftriaxone, ampicillin, streptomycin, tetracycline, sulfisoxazole, and chloramphenicol are widespread and detected in all the farms. The detection of MDR ESBL-E. coli suggests that dairy cattle can be a reservoir for these bacteria, highlighting the associated public health risk.

Genomic Profiling of Multidrug-Resistant Swine Escherichia coli and Clonal Relationship to Human Isolates in Peru.

The misuse of antibiotics is accelerating antimicrobial resistance (AMR) in Escherichia coli isolated from farm animals. The genomes of ten multidrug-resistant (MDR) E. coli isolates from pigs were analyzed to determine their sequence types, serotypes, virulence, and AMR genes (ARGs). Additionally, the relationship was evaluated adding all the available genomes of Peruvian E. coli from humans using the cgMLST + HierCC scheme. Two aEPEC O186:H11-ST29 were identified, of which H11 and ST29 are reported in aEPEC isolates from different sources. An isolate ETEC-O149:H10-ST100 was identified, considered a high-risk clone that is frequently reported in different countries as a cause of diarrhea in piglets. One ExPEC O101:H11-ST167 was identified, for which ST167 is an international high-risk clone related to urinary infections in humans. We identified many ARGs, including extended-spectrum β-lactamase genes, and one ETEC harboring the mcr-1 gene. CgMLST + HierCC analysis differentiated three clusters, and in two, the human isolates were grouped with those of swine in the same cluster. We observed that Peruvian swine MDR E. coli cluster with Peruvian E. coli isolates from healthy humans and from clinical cases, which is of great public health concern and evidence that AMR surveillance should be strengthened based on the One Health approach.

Molecular Analysis of Some Virulence Genes of Escherichia Coli Isolates from Wound and Burn Samples in Kirkuk City

Background: Gram-negative infections caused by non-spore producing E. coli that are facultatively anaerobic have been recognized as serious infections linked to higher mortality and significant costs associated with medical care.&#x0D; Aim: The goal of this study was to determine whether or not a number of E. coli strains are capable of producing virulence genes by using a technique called real time PCR.&#x0D; Materials and Methods: 155 people with burns and wounds had their cultures obtained, and susceptibility testing using swabs, tissue biopsies, and needle aspirations were performed. When compared to other techniques of specimen collection, swab cultures yield precise results and are a practicable, widely utilized, non-invasive, and cost-effective method. The two most recommended techniques for obtaining specimens are tissue biopsy and needle aspiration. Swab cultures, on the other hand, are appropriate since they are useful, widely applied, non-invasive, and reasonably priced. Samples from patients were collected in the consultation rooms, including swabs from burns and wounds. The sufferers had these taken from them. Before being identified, smear samples were initially cultivated in the lab on several selective media. Transport media-filled sterile collection containers containing these samples were brought to the lab.&#x0D; Results: According to the study's findings, E. coli was found in 27.10% of the burns and wounds samples. According to the study's findings, the majority of patients with E. coli infections (71.43 percent) were females, while only 28.57 percent were males. According to the study's findings, 28.57 percent of patients diagnosed with E. coli infections were between the ages of 31 and 40. The investigation also indicated that all of the sputum samples containing E. coli isolates were from patients above the age of 40. Furthermore, 45 percent of urine samples were collected from patients aged 31 to 40, and 42.86 percent of patients had wound infections. When examined using the MTP technique, fifty percent of the E. coli isolates tested positive for being potent biofilm producers, according to the study's findings. The DNA of 12 distinct E. coli bacterial isolates was isolated and purified using a genomic DNA purification kit. The bacteria were cultured on chromogenic agar medium to produce these E. coli isolates. The results were detected using electrophoresis on agarose gel at a concentration of 1.5 percent. The gel was then exposed to ultraviolet light, which revealed the DNA as compact bands. The virulence gene was found in 95.24 percent of the E. coli isolates taken from burn and wound sites, according to the study's findings. The majority of E. coli isolates tested positive for amikacin, imipenem, and gentamycin, according to the study's findings. The majority of E. coli isolates, on the other hand, were resistant to the medications amoxicillin, levofloxacin, cefotaxime, ceftriaxone, pefloxacin, cefepime, and ampicillin.

Study of the Genomic Characterization of Antibiotic-Resistant Escherichia Coli Isolated From Iraqi Patients with Urinary Tract Infections.

Urinary tract infection is one of the last diseases prevalent in humans, with various causative agents affecting 250 million people annually, This study analyzed UTIs in Iraqi patients caused by Escherichia coli. ESBL enzymes contribute to antibiotic resistance. The research aimed to analyze ESBL gene frequency, resistance patterns, and genetic diversity of E. coli strains; Between Dec 2020 and May 2021, 200 urine samples were collected, cultured on blood agar, EMB, and MacConkey's plates, samples incubated at 37°C for 24h. Positive samples (> 100cfu/ml) underwent Kirby-Bauer and CLSI antibiotic susceptibility testing. PCR detected virulence genes, Beta-lactamase coding genes, and biofilm-associated resistance genes in E. coli isolates; Out of 200 isolates, 80% comprised Gram-positive and Gram-negative bacteria. Specifically, 120 isolates (60%) were Gram-negative, while 40 isolates (20%) were Gram-positive. Among Gram-negative isolates, 20% were identified as E. coli. Remarkably, all E. coli strains showed resistance to all tested antibiotics, ranging from 80 to 95% resistance. The E. coli isolates harbored three identified resistance genes: blaTEM, blaSHV, and blaCTXM. Regarding biofilm production, 10% showed no formation, 12% weak formation, 62% moderate formation, and 16% strong formation; our study found that pathogenic E. coli caused 20% of UTIs. The majority of studied E. coli strains from UTI patients carried the identified virulence genes, which are vital for infection development and persistence.

Resistome and virulome of high-risk pandemic clones of multidrug-resistant extra-intestinal pathogenic Escherichia coli (ExPEC) isolated from tertiary healthcare settings in Uganda.

Multi-drug resistant (MDR) globally disseminated extraintestinal pathogenic high-risk Escherichia coli (ExPEC) clones are threatening the gains in bacterial disease management. In this study, we evaluated the genomic structure including the resistome and virulome of the E. coli isolates from extraintestinal infections using whole genome sequencing (WGS). The results highlight that isolates were highly resistant (≥ 90.0%) to commonly used antibiotics (Ampicillin, Trimethoprim-Sulfamethoxazole, Nalidixic acid, and Piperacillin) and were less (<14%) resistant to last resort antibiotics; Imipenem (10.94%) and Meropenem (10.20%). A greater proportion of the E. coli isolates belonged to phylogroup B2 (30.52%) and phylogroup A (27.37%). The sequence types ST131 of phylogroup B2 (21.05%) and ST648 of phylogroup F (9.3%) were the dominant pandemic high-risk clones identified in addition to the ST1193, ST410, ST69, ST38, ST405, and ST10. Many of the isolates were MDR and most (64.58%) carried the blaCTX-M-15 gene for extended-spectrum β-lactamases. There was a high correlation between phylogroups and the occurrence of both antimicrobial resistance and virulence genes. The cephalosporin-resistance gene blaEC-5 was only found in phylogroup B2 while blaEC-8 and blaEC-19, were only found within phylogroup D and phylogroup F respectively. Aminoglycoside gene (aadA1) was only associated with phylogroups D and C. The isolates were armed with a broad range of virulence genes including adhesins, toxins, secreted proteases, iron uptake genes, and others. The yfcv, chuA, and kpsE genes preferentially occurred among isolates of phylogroup B2. The study underlines the predominance of MDR internationally disseminated high-risk ExPEC clones with a broad range of virulence genes known to be highly transmissible in healthcare and community settings.

Trends of feline Escherichia coli minimum inhibitory concentrations over 14 years illustrate the need for judicious antimicrobial use in cats.

This study aims to assess the antimicrobial resistance (AMR) trends among Escherichia coli isolated from cats between 2008 and 2022, utilizing MIC data, within a one-health framework. The study analyzed MIC results from 1,477 feline E coli isolates that were obtained from samples submitted to the Cornell University Animal Health Diagnostic Center, primarily from the northeastern US. MIC values were categorized as susceptible or not susceptible using the Clinical and Laboratory Standards Institute breakpoints. Multidrug resistance (MDR) was analyzed using a Poisson regression model. Additionally, accelerated failure time models were employed to analyze MIC values. Out of the 1,477 E coli isolates examined, 739 (50%) showed susceptibility to all tested antimicrobials. Among the tested antimicrobials, cefazolin (69%) and ampicillin (74% for urinary tract isolates) exhibited the lowest susceptibility. Overall, 15% of isolates were not susceptible to cefovecin. E coli isolates were highly susceptible (> 95%) to antibiotics typically reserved for human use. Almost one-third of the isolates were classified as MDR, with nonurinary isolates more likely to exhibit an MDR pattern. A decrease in MICs for fluoroquinolones and gentamicin in recent years was identified. However, MICs for cephalexin increased from 2016 to 2022 and cefovecin from 2012 to 2019. This study highlights the challenge of AMR in feline medicine, emphasizing the importance of responsible antimicrobial use and surveillance to address E coli AMR. The related Currents in One Health by Cazer et al, JAVMA, December 2023, addresses additional feline antimicrobial stewardship topics.

Molecular epidemiology and pathogenomics of extended-spectrum beta-lactamase producing- Escherichia coli and - Klebsiella pneumoniae isolates from bulk tank milk in Tennessee, USA.

The rise in extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in dairy cattle farms poses a risk to human health as they can spread to humans through the food chain, including raw milk. This study was designed to determine the status, antimicrobial resistance, and pathogenic potential of ESBL-producing -E. coli and -Klebsiella spp. isolates from bulk tank milk (BTM). Thirty-three BTM samples were collected from 17 dairy farms and screened for ESBL-E. coli and -Klebsiella spp. on CHROMagar ESBL plates. All isolates were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and subjected to antimicrobial susceptibility testing and whole genome sequencing (WGS). Ten presumptive ESBL-producing bacteria, eight E. coli, and two K. pneumoniae were isolated. The prevalence of ESBL-E. coli and -K. pneumoniae in BTM was 21.2% and 6.1%, respectively. ESBL-E. coli were detected in 41.2% of the study farms. Seven of the ESBL-E. coli isolates were multidrug resistant (MDR). The two ESBL-producing K. pneumoniae isolates were resistant to ceftriaxone. Seven ESBL-E. coli strains carry the blaCTX-M gene, and five of them co-harbored blaTEM-1. ESBL-E. coli co-harbored blaCTX-M with other resistance genes, including qnrB19, tet(A), aadA1, aph(3'')-Ib, aph(6)-Id), floR, sul2, and chromosomal mutations (gyrA, gyrB, parC, parE, and pmrB). Most E. coli resistance genes were associated with mobile genetic elements, mainly plasmids. Six sequence types (STs) of E. coli were detected. All ESBL-E. coli were predicted to be pathogenic to humans. Four STs (three ST10 and ST69) were high-risk clones of E. coli. Up to 40 virulence markers were detected in all E. coli isolates. One of the K. pneumoniae was ST867; the other was novel strain. K. pneumoniae isolates carried three types of beta-lactamase genes (blaCTX-M, blaTEM-1 and blaSHV). The novel K. pneumoniae ST also carried a novel IncFII(K) plasmid ST. Detection of high-risk clones of MDR ESBL-E. coli and ESBL-K. pneumoniae in BTM indicates that raw milk could be a reservoir of potentially zoonotic ESBL-E. coli and -K. pneumoniae.