blaCTX-M Genes Research Articles

Prevalence, transmission and genomic epidemiology of mcr-1-positive colistin-resistant Escherichia coli strains isolated from international airplane waste, local resident fecal and wastewater treatment plants.

The emergence and dissemination of mcr-1-positive Escherichia coli (MCRPEC) represent a critical public health threat. Here, we conducted a prospective analysis of MCRPEC isolates from wastewater treatment plants (WWTPs), local residents' fecal (LRF), and international airplane waste (IAW) to investigate their genetic characteristics and transmission patterns circulating in human-environment domains. The MCRPEC prevalence was 2.43 % in WWTPs, 1.37 % in IAW and 0.69 % in LRF. MCRPEC showed substantial genetic diversity, encompassing 61 sequence types (primarily ST1011, ST101, and ST2705), 7 plasmid types (primarily IncI2), 8 phylogroups (primarily A and B1), 9 mcr-1-flanked lineages (primarily L5), 6 clusters (primarily C2 and C4), diverse serotypes, and 61.95 % transposon-containing strains. The mcr-1 gene co-existed with 46 antibiotic resistance genes (ARGs) and 19 virulence factor genes (VFGs). Notably, 6 IncI2 plasmids carried the blaCTX-M, IS1380, and mcr-1 genes. MCRPEC from WWTPs harbored a greater number of ARGs (56.95 ± 5.99) but fewer VFGs (15.03 ± 6.40) compared to those from human-associated sources (LRF and IAW). ST1011, ST2705, IncHI2, and L7 were prevalent in WWTP-derived MCRPEC, whereas IncX4 and L3 were more common in human-derived MCRPEC. Genetic features such as ST101, ST48, IncI2, L4, L5, C2, and C4 were simultaneously present in strains from LRF, IAW, and WWTPs. Core genetic analyses also showed genetically similar MCRPEC strains across various geographic locations. The findings underscore the extensive dissemination, strong environmental adaptation, and clonal transmission of MCRPEC across diverse reservoirs, reinforcing the urgent need for coordinated multisectoral surveillance of human and environment interfaces to effectively mitigate further transmission.

Distribution and Molecular Characterization of Antibiotic-Resistant Pseudomonas aeruginosa in Hospital Settings of Sulaymaniyah, Iraq.

Pseudomonas aeruginosa is a significant pathogen in hospital settings, notorious for its role in hospital-acquired infections and its ability to develop resistance to multiple antibiotics. This study investigates the prevalence, distribution, and antibiotic resistance gene profiles of P. aeruginosa in seven hospitals in Sulaymaniyah City. A total of 300 samples were collected from various hospital surfaces including mops, sinks, medical equipment, beds, desks, and floors. Using bacteriological, biochemical, and molecular methods, 66 isolates were confirmed as Pseudomonas species, with 26 identified as P. aeruginosa. Antibiotic susceptibility testing revealed resistance rates of 23.3% to streptomycin, 13.6% to tobramycin, 22.7% to moxifloxacin, 21.2% to levofloxacin, and 22.7% to norfloxacin. Furthermore, the antibiotic resistance gene detection showed the presence of the bla CTX-M, bla SHV, qnrB, and bla ACC-1 genes among the isolates. The study highlights a 22% contamination rate of hospital surfaces with Pseudomonas species, emphasizing the urgent need for enhanced infection control measures and targeted antimicrobial stewardship to manage and reduce the spread of multidrug-resistant P. aeruginosa.

Enterobacterales Producing ESBLs and AmpC in Fresh Vegetables from Tebessa City, Algeria.

This study aimed to evaluate the contamination levels of fresh products by ESBLs-producing Enterobacterales (ESBLs-E) or AmpC-producing Enterobacterales and characterize ESBLs genes. A total of 132 samples (67 vegetables and 65 fruits) were collected from markets in Tebessa, eastern Algeria. Among the samples, 16 third-generation cephalosporin-resistant Enterobacterales isolates were identified with a prevalence of 19.40% in vegetable samples, while there was no positive finding in fruit samples. Isolates showed resistance to most β-lactams, and all of them displayed multidrug resistance. Phenotypic tests for ESBLs detection, using double-disk synergy test and double-disk test were positive for 14 strains, including Klebsiella pneumoniae (n = 5), Klebsiella oxytoca (n = 4), Klebsiella terrigena (n = 2), Kluyvera spp. (n = 2), and Enterobacter cloacae (n = 1). Two AmpC-producing strains (Citrobacter freundii and E. cloacae) were identified through the AmpC disk test. Contamination rates of vegetables by ESBLs-E and AmpC-producing Enterobacterales were 19.40% and 2.98%, respectively. PCR results showed the presence of at least one ESBL gene in seven selected strains, with the dominance of blaCTX-M gene. Notably, K. pneumoniae strains showed the co-occurrence of two or three genes. Sequencing identified uncommon variants of ESBLs genes for the first time in Algeria, including blaCTX-M-79 (2/7), blaCTX-M-107 (2/7), blaCTX-M-117 (2/7), blaTEM-112 (1/7), blaTEM-125 (2/7), blaTEM-194 (1/7), and blaSHV-176 (3/7).

Prevalence of Resistance Genes Among Multidrug-Resistant Gram-Negative Bacteria Isolated from Waters of Rivers Swat and Kabul, Pakistan.

The waters of rivers Swat and Kabul are the main water source for domestic and irrigation purposes in the northwestern part of Pakistan. However, this water has been contaminated due to human activities. This study aimed to analyze the water of these rivers for occurrence of antibiotic resistance genes among Gram-negative bacteria. Samples were collected from 10 different locations of these rivers. The samples were processed for the isolation of Gram-negative bacteria. Isolated bacteria were checked against 12 different antibiotics for susceptibility. The isolates were also analyzed for the presence of seven antibiotic resistance genes. A total of 50 bacterial isolates were recovered that belonged to five different bacterial genera, that is, Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa, Raoultella terrigena (Klebsiella terrigena), and Pseudomonas fluorescens. Antibiotic resistance pattern was cefixime 72%, cephalothin 72%, ampicillin 68%, nalidixic acid 68%, kanamycin 54%, streptomycin 42%, sulfamethoxazole-trimethoprim 28%, chloramphenicol 28%, meropenem 8%, gentamicin 8%, amikacin 2%, and tobramycin 2%. The prevalence of bla-TEM gene was 72% (n = 36), aadA gene 34% (n = 17), sul gene 32% (n = 16), bla-CTXM gene 12% (n = 6), int gene 66% (n = 33), and int1 gene 6% (n = 3). This information highlights the need for controlling and monitoring the release of domestic wastes to rivers.

Lack of wastewater treatment in a small town drives the spread of ESBL-producing Escherichia coli in irrigation waters.

Antibiotic resistance (ABR) is a critical and growing global challenge, especially in low- and middle-income countries. Ecuador has made great progress in connecting households to piped water supplies; however, the collection and treatment of domestic wastewater has lagged. This infrastructural gap may be accelerating the spread of ABR into surface waters used downstream for irrigation. We studied the contributions of a small town in Ecuador to the prevalence of extended-spectrum β-lactamase-producing Escherichia coli in a glacial stream used for irrigating crops. The study analyzed water samples upstream (n=60) and downstream (n=60) of the town of Píntag as well as 30 lettuce samples irrigated by surface waters downstream of the town. A subset of third generation cephalosporin resistant E. coli (3GCR-EC) isolates (n=58) were sequenced to characterize antibiotic resistance genes and pathogenic lineages. Our results showed that there was nearly a three-log increase in mean E. coli colony forming units in the downstream samples versus upstream. At the upstream sites above the town of Píntag, 6.7% of water samples were positive for 3GCR-EC compared to 100% of samples collected at the downstream sites. Additionally, 70.1% of sequenced 3GCR-EC isolates collected at downstream sites carried blaCTX-M genes and 3.4% belonged to pandemic lineages ST131 and ST10. As countries develop household piped water infrastructure, attention should focus on how the lack of domestic wastewater collection and treatment may accelerate the spread of ABR in waterways and the food system.

Detecting and Reporting Extended Spectrum Beta-Lactamases Producers: Evaluation of Three Approaches and the Clinical Impact

Abstract Introduction/Objective Since the COVID-19 pandemic, bacteria producing extended spectrum beta-lactamases (ESBLs) have increased >30%. Laboratories can perform phenotypic microbroth dilution or molecular approaches to identify ESBL producers. However, the gold standard is a laborious double disc synergy test. We evaluated the concordance among the three approaches, the clinical impact, and justification of reporting ESBL producers. Methods/Case Report Seventy-eight ESBL producers identified by microbroth dilution (MicroScan WalkAway, Beckman Coulter, Brea, California, USA) or detection of blaCTX-M gene from blood cultures (BCID2 Panel, BioFire, Biomerieux, Salt Lake City, Utah, USA) were tested using double disc synergy test retrospectively. Concordance among the methods evaluated. Discordant cases were reviewed with subsequent patient chart review. Results (if a Case Study enter NA) Tested isolates included E. coli (n=44, 57%), K. pneumoniae (n=19, 24%), K. oxytoca (n=7, 9%), and P. mirabilis (n=8, 10%). Concordance between microbroth dilution and disk diffusion was 86% (67/78). For isolates with genotypic testing (n=16), concordance with microbroth dilution and disk diffusion was 93% and 88%, respectively. Overall concordance was 88%. There were 12 isolates (E. coli:8, P. mirabilis: 2, and K. pneumoniae:2) with discordant results from one method. The susceptibility profile of discordant isolates revealed resistance to ceftriaxone or ceftazidime (11/12), penicillin (11/12), and aztreonam (6/12); 10/12 isolates were resistant to at least 2 groups of these antibiotics, a phenotype of ESBL producers. Clinical teams acknowledged presence of ESBLs in 83% (10/12) of cases and 58% (7/12) warranted treatment with meropenem or combinational therapy. Infection prevention implemented contact isolate procedures in all cases. Conclusion Our current in-house methods (e.g., microbroth dilution and genotypic testing) are still justified for ESBL testing to optimize patient care in a timely fashion. In the minor cases with discordant results, clinical care was unaffected. Laboratory workflow changes need to take laboratory efficiency, patient outcomes, and actionable clinical changes into consideration.

Isolation and molecular characterization of multi-drug resistant uropathogenic Escherichia coli from urine samples: Insights into urinary tract infection management

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) present a significant global health burden. With UPEC responsible for approximately 90 % of UTIs, understanding its mechanisms of virulence and antibiotic resistance is paramount for effective treatment strategies. This study aimed to investigate the prevalence of multi-drug resistant (MDR) E. coli strains in urinary tract infections and to elucidate the molecular mechanisms underlying their resistance. A total of 150 E. coli isolates were obtained from urinary tract infection specimens collected patients at Doctor’s Diagnostic Centre in Kalyan Nagar, Bangalore over four months November 2022 to February 2023. Isolation and screening of pathogenic E. coli were conducted using Eosin Methylene Blue Agar and Nutrient Agar, followed by identification through gram staining and biochemical characterization tests. Antibiotic susceptibility testing was performed using Mueller-Hinton agar and various antibiotic discs. Phenotypic and molecular identification of extended-spectrum beta-lactamase (ESBL) producers were conducted, along with the screening for specific antibiotic resistance genes using polymerase chain reaction (PCR). Of the 150 E. coli isolates, 57.3 % were found to be multi-drug resistant, with all 35 isolates from hospitalized patients exhibiting MDR characteristics. Phenotypic and molecular analyses revealed the presence of ESBL-producing isolates, with the blaCTX-M, blaTEM, and blaSHV genes detected in PCR assays. Plasmid extraction and PCR assays further identified the presence of plasmid-encoded carbapenemase genes (blaOXA-48, blaNDM-1, blaKPC-2), as well as other antibiotic resistance genes including those conferring resistance to fluoroquinolones. The results underscore the alarming prevalence of MDR E. coli strains in urinary tract infections, particularly in hospitalized patients. Molecular characterization revealed the diverse genetic mechanisms contributing to antibiotic resistance, including the presence of ESBLs and plasmid-encoded resistance genes. These findings emphasize the urgent need for surveillance and infection control measures to mitigate the spread of MDR pathogens and the development of alternative therapeutic strategies to combat antibiotic resistance in UTIs.

Detection of extended-spectrum beta-lactamase genes among Escherichia coli isolates of buffalo mastitis milk

In recent years, bacteria carrying extended-spectrum beta-lactamase (ESBL) genes have become increasingly prevalent. These genes provide resistance to antibiotics, making treatment more challenging. Escherichia coli is a major cause of mastitis in buffaloes, and there have been reports of ESBL genes in E. coli isolated from bovine mastitis milk. This study was aimed to detect ESBL genes in E. coli found in mastitis buffalo milk. A total of 100 samples of mastitis milk were collected from different buffalo farms in Karachi for gene detection and antibiotic sensitivity evaluation. Among the samples, 44 % were positive for E. coli, confirmed by specific characteristics and biochemical properties. To check the antimicrobial sensitivity of isolates MIC test was carried out using micro broth dilution method. Antibiotic sensitivity tests revealed that out of the 44 isolates, 68 % were resistant to ampicillin, 81 % to tetracycline, 52 % to levofloxacin, 27 % to cefoxitin, and 25 % to ceftriaxone at various concentrations. Additionally, 7 % of the isolates showed resistance to all classes of antibiotics tested. PCR results indicated that 25 % of the total isolates carried the blaCTX-M-1 ESBL gene, while no other types of blaCTX-M, blaSHV, or blaTEM genes were detected. It was discovered that multidrug resistance and the presence of the blaCTX-M-1 ESBL gene in E. coli isolates from mastitis milk pose a significant threat to veterinarians and human clinicians, as they are highly resistant to beta-lactams and other commonly used antibiotics for mastitis treatment.

Temporal dynamics of extended-spectrum β-lactamase-producing Escherichia coli and carbapenemase-producing Gram-negative bacteria in hospital wastewater

Hospital wastewater is a reservoir for the environmental spread of clinically relevant antimicrobial-resistant bacteria and resistance genes. The aim of this study was to quantify total Escherichia coli, extended-spectrum β-lactamase (ESBL)-producing E. coli, and carbapenemase-producing organisms (CPOs) and perform whole-genome sequencing-based characterization of these bacterial isolates in hospital wastewater samples collected bimonthly in Japan from January to November 2021. Total E. coli counts were 8.1 × 103–8.8 × 104 colony-forming units (CFU)/mL. ESBL-producing E. coli were detected in the sampling months of January, March, May, and July, with the ratio of ESBL-producing E. coli to total E. coli being remarkably highest (95 %) in July. In contrast, DHA-1 Ambler class C β-lactamase (AmpC)-producing E. coli was detected in September and November, accounting for 28 % and 3 % of total E. coli counts, respectively. All 140 ESBL-producing E. coli isolates harbored the blaCTX-M genes, with blaCTX-M-14 being the most common genotype (94.3 %), the vast majority of which were associated with the human virulent B2-O25b: H4-ST131-fimH30R/non-Rx. In September, E. coli clade I-O8:H33-ST3910-fimH1074 was primarily associated with blaDHA-1. Among 26 representative CPO isolates, Aeromonas caviae (34.6 %) and A. hydrophila subsp. hydrophila (30.8 %) were dominant. The most frequently detected carbapenemase gene was blaIMP-1 (57.7 %), followed by blaGES-24 (34.6 %) and blaGES-4 (7.7 %). Estimated bacterial counts of CPOs ranged from 4.0 × 10−1 to 4.7 × 103 CFU/mL over the six sampling months. blaIMP-1-positive A. hydrophila subsp. hydrophila ST860, which was repeatedly detected over the five sampling months, accounted for the highest total number of this bacterial clone (79 %).Overall, this study provides insights into the overwhelming presence and persistence of E. coli B2-O25b:H4-ST131-H30R/non-Rx with blaCTX-M-14 and Aeromonas spp. with blaIMP-1 in hospital wastewater, and the change in the dynamics of resistance gene prevalence from blaCTX-M-positive E. coli to blaDHA-1-positive E. coli.

Molecular characterization of blaTEM and blaCTX-M ESBLs genes producing Escherichia coli isolates from urinary tract infections (UTIs) in Al-Basrah province, Iraq

Urinary tract infections (UTIs) are primarily caused by Gram-negative Enterobacteriaceae bacteria, accounting for 80–90% of uropathogenic Escherichia coli infections. Multidrug-resistant (MDR) bacteria, such as extended-spectrum β-lactamases (ESBLS), pose a significant threat to global healthcare systems. The current study relied on 200 urine samples collected from patients suffering urinary tract infections (UTIs) in Al Sadr Teaching Hospital in Al-Basrah Province,Iraq from the 5th January to 22nd February. A result of cultivated 200 samples collected from urinary tract infection patients showed 71 (35.5%) positive samples on MacConkey agar, eosin methylene blue (EMB), and HiChromeTM E.coli agar, and bacterial growth was distributed to 47 (66.2%) Escherichia coli and 24 (33.8%) of Gram-negative species. The diagnostic gene 16S rDNA by PCR method showed that all 47 E. coli isolates had a molecular weight of around 585 bp at 100%. Furthermore, positive results were shown in 44 (93.6%) of the E.coli isolates in the current study that produced ESBLs by using the double-disc approximation method (DAM). While 3 (6.4%) isolates revealed negative results for ESBLs, whereas the13(27.7%) of the E.coli isolates gave positive results for ESBL. While 34(72.3%) E.coli isolates showed negative results for produced ESBLs by using the double-disc synergy test (DDST), there were significant differences (P<0.01) between positive and negative isolates in both methods used to detect ESBL-producing isolates. Extended-spectrum β-lactamses (ESBLs) genes were detected by using PCR to amplify the blaTEM and blaCTX-M genes in E.coli isolates. The amplified genes' bands were characterised approximately at (800 bp) for blaTEM and (754 bp) for blaCTX-M , the products were compared to the standard molecular DNA ladder at (2000 bp). According to the results, 47(100%) E.coli isolates gave positive results for the blaTEM gene. While blaCTX-M gene was shown, only 23(48.9%) E.coli isolates had positive results for the blaCTX-M gene.

Molecular Characterization of Carbapenem and Colistin Resistance in Klebsiella pneumoniae Isolates Obtained from Clinical Samples at a University Hospital Center in Algeria.

The current study aimed to determine the molecular mechanisms of carbapenem and colistin resistance among the clinical isolates of Klebsiella pneumoniae from hospitalized patients admitted to a university hospital in Eastern Algeria. In total, 124 non-duplicate isolates of K. pneumoniae were collected from September 2018 to April 2019. Bacterial identification was performed using MALDI-TOF MS. The presence of extended spectrum β-lactamase (ESBL) genes, carbapenemase genes, chromosomal mutation and mcr genes in colistin-resistant K. pneumoniae were evaluated by PCR. ESBLs represented a rate of 49.1% and harbored blaCTX-M, blaTEM and blaSHV genes. Concerning carbapenems, 12 strains (9.6%) were resistant to ertapenem (MIC: 1-32 μg/mL), of which one strain (0.8%) was also resistant to imipenem (MIC: 32 μg/mL). Among these strains, nine (75%) harbored blaOXA-48 gene. Seven strains (5.6%) expressed resistance to colistin (MIC: 2-32 μg/mL), of which two harbored mcr-8 and mgrB genes simultaneously. The existence of a double resistance to colistin in the same strain is new in Algeria, and this could raise concerns about the increase in levels of resistance to this antibiotic (MIC: 32 μg/mL). The mgrB gene alone was observed in five isolates (71.4%), including two strains harboring blaOXA-48. This is the first report revealing the presence of K. pneumoniae strains carrying the blaOXA-48 gene as well as a mutation in the mgrB gene. Large-scale surveillance and effective infection control measures are also urgently needed to prevent the outbreak of various carbapenem- and colistin-resistant isolates.

Phenotypic and Genotypic Profiles of Extended-Spectrum Beta-Lactamase-Producing Multidrug-Resistant Klebsiella pneumoniae in Northeastern Thailand.

The global emergence of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae presents a significant public health threat and complicates antibiotic treatment for infections. This study aimed to determine the prevalence of ESBL-producing K. pneumoniae in a clinical setting, analyze their antimicrobial susceptibility profiles, and characterize both phenotypic and genetic determinants. A total of 507 non-duplicate clinical isolates of Enterobacterales were collected between 2019 and 2020, and third-generation cephalosporin resistance was screened by disk diffusion. Identification of K. pneumoniae was confirmed using biochemical tests and PCR with species-specific primers. Antimicrobial susceptibility testing was conducted using disk diffusion, and phenotypic ESBL production was confirmed using the combined disk method. Multiplex PCR detected ESBL genes (blaTEM, blaSHV, and blaCTX-M) and identified blaCTX-M groups. The genetic relatedness of ESBL-producing strains was assessed using the ERIC-PCR approach. Fitty-four isolates were confirmed as ESBL producers, all classified as multidrug-resistant (MDR). All ESBL-producing K. pneumoniae isolates exhibited resistance to ampicillin and cefotaxime, with high resistance rates for ciprofloxacin (98.2%), azithromycin (94.4%), piperacillin-tazobactam (88.9%), and trimethoprim (83.3%). Genotypic analysis revealed blaCTX-M was present in 94.4% of isolates, blaSHV in 87%, and blaTEM in 55.5%. The blaCTX-M-1 group was the most prevalent, accounting for 96.1% of isolates. Co-harboring of blaCTX-M, blaSHV, and blaTEM occurred in 42.6% of isolates, with co-carrying of blaCTX-M, and blaSHV was observed in 23/54 isolates. The ERIC-PCR analysis revealed 15 distinct types, indicating high genetic diversity. These findings highlight the urgent need for ongoing monitoring to control the spread of ESBL among K. pneumoniae and emphasize the importance of early detection and appropriate antibiotic selection for effectively treating infection caused by these pathogens.

Prevalence and molecular characterization of ESBL/pAmpC producing faecal Escherichia coli strains with widespread detection of CTX-M-15 isolated from healthy poultry flocks in Eastern Algeria

The intensification of livestock farming has led to the widespread use of massive amounts of antibiotics worldwide. Poultry production, including white meat, eggs and the use of their manure as fertiliser, has been identified as one of the most crucial reservoirs for the emergence and spread of resistant bacteria, including E. coli in poultry as an important opportunistic pathogen representing the greatest biological hazard to human and wildlife health. Thus, this study aimed to analyse E. coli in the faecal carriage of healthy poultry flocks and to investigate the phenotypic and genotypic characteristics of antimicrobial resistance, including integrons genes and phylogenetic groups. A total of 431 cloacal swabs from apparently healthy poultry from four regions in Eastern Algeria from December 2021 to October 2022. 360 E. coli were isolated; from broilers (n = 151), broiler breeders (n = 91), laying hens (n = 72), and breeding hens (n = 46). Among this, 281 isolates exhibited multidrug resistance (MDR) phenotype, 17 of the 360 E. coli isolates exhibited ESBL, and one isolate exhibited both ESBL/pAmpC. A representative collection of 183 among 281 MDR E. coli was selected for further analysis by PCR to detect genes encoding resistance to different antibiotics, and sequencing was performed on all positive PCR products of blaCTX-M and blaCMY-2 genes. Phylogenetic groups were determined in 80 E. coli isolates (20 from each of the four kinds of poultry). The blaCTX-M gene was found in 16 (94.11 %) ESBL-producing E. coli isolates within 11 strains co-expressing the blaSHV gene and 8 strains co-expressing the blaTEM gene. Sequence analysis showed frequent diversity in CTX-M-group-1, with blaCTX-M-15 being the most predominant (n = 11), followed by blaCTX-M-1 (n = 5). The blaCMY-2 gene was detected only in one ESBL/pAmpC isolate. Among the 183 tested isolates, various antimicrobial resistance genes were found (number of strains) blaTEM (n = 121), blaSHV (n = 12), tetA (n = 100), tetB (n = 29), sul1(n = 67), sul2 (n = 32), qnrS (n = 45), qnrB (n = 10), qnrA (n = 1), catA1(n = 13), aac-(6′)-Ib (n = 3). Furthermore, class 1 and class 2 integrons were found in 113 and 2 E. coli, respectively. The isolates were classified into multiple phylogroups, including A (35 %), B1 (27.5 %), B2 and D each (18.75 %). The detection of integrons and different classes of resistance genes in the faecal carriage of healthy poultry production indicates that commensal E. coli could potentially act as a reservoir for antimicrobial resistance, posing a significant One Health challenge encompassing the interconnected domains of human, animal health and the environment. Here, we present the first investigation to describe the diversity of blaCTX-M producing E. coli isolates with widespread detection of CTX-M-15 and CTX-M-1 in healthy breeders (Broiler and breeding hens) in Eastern Algeria.

Association of antibiotic resistance and biofilm formation in Escherichia coli ST131/O25b.

Urinary tract infections are becoming difficult to treat every year due to antibiotic resistance. Uropathogenic Escherichia coli (UPEC) isolates pose a threat with a combined expression of multidrug-resistance and biofilm formation. ST131 clone is a high-risk pandemic clone due to its strong association with antimicrobial resistance, which has been reported frequently in recent years. This study aims to define risk factors, clinical outcomes, and bacterial genetics associated with ST131/O25b UPEC. In this study, antibiotic susceptibility and species-level identification of 61 clinical E. coli strains were determined by automated systems. Detection of extended-spectrum beta-lactamases was assessed by double-disk synergy test. Biofilm formation was quantified by spectrophotometric method. Virulence genes (iutA, sfa cnf-1, iroN, afa, papA, fimA), antibiotic resistance genes (blaCTX-M, blaTEM, blaSHV, blaOXA, qnrA, qnrB, qnrS, ant(2')-Ia, ant(3)-Ia, aac(3)-IIa, mcr-1, mcr-2, mcr-3, mcr-4) were investigated by PCR. The following beta-lactamase genes were identified, blaTEM (n = 53, 86.8%), blaCTX-M (n = 59, 96.7%), blaSHV (n = 47, 77.0%), and blaOXA-1 (n = 27, 44.2%). Our data revealed that 93.4% of (57/61) E. coli isolates were biofilm-producers. O25pabBspe and trpA2 were investigated for the presence of ST131/O25b clone. Among multidrug resistant isolates, co-existence of O25pabBspe and trpA2 was detected in 29 isolates (47.5%). The fimH30 and H30Rx subclones were detected in four isolates that are strong biofilm-producers. These results suggest that clinical E. coli strains may become reservoirs of virulence and antibiotic resistance genes. This study demonstrates a significant difference in biofilm formation between E. coli ST131 and non-ST131 isolates. Moreover, 86.21% (n = 25) of ST131 isolates produced strong to moderate biofilms, while only 43.75% (n = 14) of non-ST131 isolates showed the ability to form strong biofilms. Presence of iutA and fimA genes in the majority of ST131 strains showed an important role in biofilm formation. These findings suggest application of iutA and fimA gene suppressors in treatment of infections caused by biofilm-producing drug-resistant ST131 strains.

Prevalence and characteristics of ESBL-producing Salmonella in Weifang, China.

This study examined the prevalence and antibiotic resistance pattern of blaCTX-M extended-spectrum β-lactamase positive Salmonella species isolated from a hospital in Weifang. Salmonella strains were isolated from hospitalized patients from January 2018 to April 2023. Whole-genome sequencing was performed by Illumina platform. CTX-M-producing Salmonella were identified by Comprehensive Antibiotic Research Database (CARD). Strain susceptibility to six antimicrobial agents was assessed by BD Phoenix™ M50 System. MLST analysis confirmed sequence types and additionally, serotypes were determined by SeqSero2. Genetic environments of blaCTX-M genes were analyzed by Isfinder and BLASTn. Single nucleotide polymorphisms were used to construct a phylogenetic tree to analyze homology.A total of 34 CTX-M-producing Salmonella were detected. The most prevalent serotype was Salmonella enterica subsp. enterica 1,4,[5],12:i:- (14/34, 41.18%), belonging to ST34, followed by SalmonellaEnteritidis (10/34, 29.41%), belonging to ST11. The highest resistance rate was detected to ampicillin (97.06%), followed by ceftriaxone (94.12%) and ceftazidime (58.83%). In CTX-M-producing Salmonella five types of blaCTX-M genes were identified, the most prevalent was blaCTX-M-55 (47.06%, 16/34), followed by blaCTX-M-14, blaCTX-M-65, blaCTX-M-125, and blaCTX-M-27 at 26.47% (9/34), 11.77% (4/34), 8.82% (3/34), and 5.88% (2/34), respectively. Apart from blaCTX-M, 40 antibiotic resistance genes were also detected, conveying resistance to multiple drugs and the most frequent genes were namely, mcr-1.1, aph(6)-Id, aph(3″)-Ib, oqxAB, qnrB6, qnrS1. According to genetic environment analysis, the insertion sequence ISEcp1 was prevalent upstream of the blaCTX-M gene. Our study demonstrates that multiple resistance genes are carried by clinical isolates of Salmonella spp. however, the dominant ESBL genotype is CTX-M-55, that is associated with ISEcp1.

Detection of extended-spectrum β-lactamase producing Escherichia coli in table eggs from Istanbul.

ESBL-producing Escherichia coli strains threaten public health and obligate the use of last-resort antibiotics. This study identified 15 E. coli isolates through 16S rRNA and gyrB genes, specific to E. coli, in 120 egg samples (12.5%). Antibiotic resistance was detected according to the EUCAST and CLSI in E. coli isolates. 2 isolates were susceptible to all antibiotics, one isolate was resistant to one antibiotic, one isolate was resistant to 2 antibiotics, and 11 E. coli isolates (73.3%) had multidrug resistance. Most frequent antibiotic resistances were detected against ampicillin (80%), tetracycline (66.6%), and chloramphenicol (66.6%). A double-disc confirmation test was used to detect ESBL production, and blaTEM, blaSHV, blaCTX-M and blaOXA genes were searched by PCR. The blaTEM (100%) gene was found in all resistant E. coli isolates, and the blaCTX-M gene was detected in only 3 (20%) E. coli isolates. None of the E. coli isolates contained the genes responsible for carbapenem and colistin resistance. Our results show that multi-drug antibiotic resistance and the blaTEM gene are frequent in E. coli from table eggs in Istanbul. This is the first preliminary study on ESBL-producing E. coli isolates in table eggs in Türkiye.

Antimicrobial Susceptibility of Commensal Escherichia coli from Pig Fecal Samples and Enhanced Sensitivity for Direct Detection of the blaCTX-M Gene by Nested PCR.

The commensal Escherichia coli in the gut of pigs is a major reservoir of antimicrobial resistance and can result in possible transmission to humans through the food chain. Direct detection of E. coli from fecal samples is challenging and can be used as a bioindicator of antimicrobial resistance. This study aimed to compare the antimicrobial susceptibility profiles in commensal E. coli from antibiotic- and nonantibiotic-using pig farms and developed the direct detection of ESBL genes in pig fecal samples using nested PCR (nPCR) and multiplex PCR (mPCR) techniques. All direct genotypic results were validated with the results of PCR sequencing of isolated E. coli colonies. The ESBL-producing E. coli were found in 98.6% (145 isolates) and 96.6% (144 isolates) of antibiotic-using and nonantibiotic-using farms, respectively, predominantly CTX-M-55. The nPCR decreased the limit of detection (LOD) from sPCR about 100 times, and the lower LODs of 102, 101, and 1 CFU/mL were reached after incubating samples in an enrichment medium for 2, 4, and 8 h, respectively. The mPCR, sPCR, and nPCR techniques showed sensitivities of 30.15%, 69.85%, and 91.91%, respectively, compared to PCR sequencing. The stability and recycling of ESBL genes were independent of antibiotic usage in commensal E. coli originating in pig farms.

Genomic insights into epidemic plasmids carrying bla CTX-M and mcr-1 genes in Escherichia coli from Lebanese broiler production.

In a previous nationwide survey in the Lebanese broiler production, multidrug-resistant CTX-M-producing E. coli were found to carry the mobile colistin resistance gene mcr-1. To investigate the mobile genetic supports responsible for the spread of these resistance genes among E. coli in healthy broilers in Lebanon. Thirty-three bla CTX-M and mcr-1 positive E. coli of various sequence types from 17 broilers farms were subjected to conjugation assays. Long-read sequencing (Oxford Nanopore Technologies) and hybrid assembly were performed to determine complete plasmid sequences and their phylogenetic diversity. Twenty-nine conjugative IncFII plasmids harboured the extended-spectrum β-lactamase genes bla CTX-M-3 (n = 25) or bla CTX-M-55 (n = 4). Highly related IncF2:A-:B-/bla CTX-M-3 plasmids differing only through IS-mediated genetic rearrangements in antibiotic resistance gene clusters were found in genetically diverse E. coli strains isolated from distant farms. The mobile colistin resistance genes mcr-1.1 and mcr-1.26 were carried by IncX4 and IncI2 plasmids. Worryingly, in one isolate, the ISEcp1-bla CTX-M-55 transposable unit was found integrated in a mcr-1.26-carrying IncX4 plasmid. Beside expanded cephalosporins and colistin resistances, all E. coli isolates were multidrug-resistant with different additional resistances against aminoglycosides, (fluoro)quinolones, fosfomycin, phenicols, sulphonamides, tetracycline and trimethoprim. Closely related blaCTX-M-3/55-borne IncF2:A-:B- plasmids harbouring variable MDR regions and mcr-1 carrying IncX4 plasmids are widely disseminated in the E. coli population of healthy broilers in Lebanon. Further surveillance programmes of antimicrobial resistance and interventions to reduce the abusive use of medically important antibiotics are necessary to limit the spread of resistances in food-producing animals in Lebanon.

Presence of extended-spectrum beta-lactamase-producing Escherichia coli and carbapenem resistance in ready-to-eat stuffed mussels in Istanbul

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

Prevalence of plasmid-mediated quinolone resistance genes in extended-spectrum beta-lactamase producing Klebsiella pneumoniae isolates in northern Iran

Plasmid-mediated quinolone resistance (PMQR) in extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae (K. pneumoniae) contributes to treatment failures, extended hospital stays, and increased mortality percentages. We aimed to determine the prevalence of PMQR genes in ESBL-producing K. pneumoniae isolates from clinical samples in Babol, North of Iran region. This is the first study in this region to investigate this specific association. A total of 95 K. pneumoniae isolates were obtained from hospitalized patients with various clinical infections during March 2022 to February 2023. Disk diffusion and Combination disk method were performed to identification of antimicrobial resistance profiles and ESBL-producing strains. The presence of ESBL and PMQR genes among K. pneumoniae isolates was assessed using polymerase chain reaction (PCR) method. Of the isolates, 68 (71.57 %) were considered as ESBL-producers. The blaTEM, blaSHV and blaCTX-M genes were detected in 74.73 %, 57.89 %, and 41.05 % of K. pneumoniae isolates, respectively. Among the PMQR encoding genes, the highest and lowest frequency was associated to qepA (67.3 %) and qnrA (4.2 %), respectively. The frequency of qnrA, qnrB, qnrS, acc (6′)-Ib-cr, qepA, oqxA, and oqxB genes in 26 MDR-Kp isolates was 11.53 % (n; 3), 69.23 % (n; 18), 65.38 % (n; 17), 73.07 % (n; 19), 80.76 % (n; 21), 84.61 % (n; 22), and 76.92 % (n; 20), respectively. Our result revealed of the 68 ESBL gene-positive isolates, 60 (88.23 %) were positive for the PMQR gene. The co-occurrence of these genes within resistant isolates suggests potential linkage on mobile genetic elements such as plasmids. These findings highlight the significant burden of PMQR determinants in ESBL-producing K. pneumoniae and underscore the urgent need for effective control measures. Implementing robust antimicrobial stewardship programs and strengthening drug-resistance surveillance and control protocols are crucial to prevent the spread of resistant isolates.