Beta-lactamase Gene Research Articles

Occurrence of Extended-Spectrum Beta-Lactamase and Quinolone Resistance Genes among Escherichia coli and Klebsiella pneumoniae Isolated from Poultry, Domestic Pigs and Environ in Msimbazi River Basin in Tanzania

Occurrence of Extended-Spectrum Beta-Lactamase and Quinolone Resistance Genes among Escherichia coli and Klebsiella pneumoniae Isolated from Poultry, Domestic Pigs and Environ in Msimbazi River Basin in Tanzania

Genomic landscape of NDM-1 producing multidrug-resistant Providencia stuartii causing burn wound infections in Bangladesh

The increasing antimicrobial resistance in Providencia stuartii (P. stuartii) worldwide, particularly concerning for immunocompromised and burn patients, has raised concern in Bangladesh, where the significance of this infectious opportunistic pathogen had been previously overlooked, prompting a need for investigation. The two strains of P. stuartii (P. stuartii SHNIBPS63 and P. stuartii SHNIBPS71) isolated from wound swab of two critically injured burn patients were found to be multidrug-resistant and P. stuartii SHNIBPS63 showed resistance to all the 22 antibiotics tested as well as revealed the co-existence of blaVEB-6 (Class A), blaNDM-1 (Class B), blaOXA-10 (Class D) beta lactamase genes. Complete resistance to carbapenems through the production of NDM-1, is indicative of an alarming situation as carbapenems are considered to be the last line antibiotic to combat this pathogen. Both isolates displayed strong biofilm-forming abilities and exhibited resistance to copper, zinc, and iron, in addition to carrying multiple genes associated with metal resistance and the formation of biofilms. The study also encompassed a pangenome analysis utilizing a dataset of eighty-six publicly available P. stuartii genomes (n = 86), revealing evidence of an open or expanding pangenome for P. stuartii. Also, an extensive genome-wide analysis of all the P. stuartii genomes revealed a concerning global prevalence of diverse antimicrobial resistance genes, with a particular alarm raised over the abundance of carbapenem resistance gene blaNDM-1. Additionally, this study highlighted the notable genetic diversity within P. stuartii, significant informations about phylogenomic relationships and ancestry, as well as potential for cross-species transmission, raising important implications for public health and microbial adaptation across different environments.

Polymyxin Resistance in Salmonella: Exploring Mutations and Genetic Determinants of Non-Human Isolates.

Until 2015, polymyxin resistance was primarily attributed to chromosomal mutations. However, with the first report of mobile colistin resistance (mcr-1) in commensal Escherichia coli from food animals in China, the landscape has changed. To evaluate the presence of polymyxin resistance in Salmonella spp., a drop screening test for colistin and polymyxin B was carried out on 1156 isolates of non-human origin (animals, food, and the environment), received in Brazil, between 2016 and 2021. Subsequently, 210 isolates with resistant results in the drop test were subjected to the gold-standard test (broth microdilution) for both colistin and polymyxin B. Whole-genome sequencing (WGS) of 102 resistant isolates was performed for a comprehensive analysis of associated genes. Surprisingly, none of the isolates resistant to colistin in the drop test harbored any of the mcr variants (mcr-1 to mcr-10). WGS identified that the most common mutations were found in pmrA (n= 22; T89S) and pmrB (n = 24; M15T, G73S, V74I, I83A, A111V). Other resistance determinants were also detected, such as the aac(6')-Iaa gene in 72 isolates, while others carried beta-lactamase genes (blaTEM-1blaCTX-M-2, blaCMY-2). Additionally, genes associated with fluoroquinolone resistance (qnrB19, qnrS1, oqxA/B) were detected in 11 isolates. Colistin and polymyxin B resistance were identified among Salmonella from non-human sources, but not associated with the mcr genes. Furthermore, the already-described mutations associated with polymyxin resistance were detected in only a small number of isolates, underscoring the need to explore and characterize unknown genes that contribute to resistance.

Antibiotic resistance profile in sediments and bacterial isolates from a human-impacted mangrove from Bahía Magdalena, Mexico

Abstract Antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) in the environment are considered emerging contaminants. One of the ecosystems that can harbor resistant bacteria and genes is mangroves. This report aims to assess ARB and ARG's presence at two mangrove locations in Bahia Magdalena, Mexico. Bacterial isolates were selected based on their phenotypic profiles of antibiotic resistance. DNA from mangrove sediments and bacterial isolates was tested for int1 and antibiotic resistance genes by final point PCR. From 90 ARB isolates, the phenotypic profile showed resistance to ciprofloxacin (90%), cefotaxime (85.7%), and ampicillin (78.6%), while the beta-lactamase gene (blaCTX-M) was present in 72.9% of the isolates. Antibiotic resistance genes identified in sediments were found even at 30–50 cm depth, but the integron Class gen (int1) was found mainly in shallow samples. Principal component analysis showed a close relationship between the integron gene, vancomycin, and sulfonamides resistance genes. No correlation was found between the phenotypic and genotypic profiles of the bacterial isolates. Also, no differences were observed between the two mangrove ecosystems studied. The impact of anthropogenic activities was equally significant on both sites. The role of mangrove ecosystems in the dispersion of antibiotic resistance needs to be further explored.

PREVALENCE AND CHARACTERISATION OF EXTENDED-SPECTRUM BETA-LACTAMASES AND PLASMID-MEDIATED QUINOLONES RESISTANCE IN Enterobacteriaceae ISOLATED FROM COMPANION ANIMALS

Antimicrobial resistance is a major public health concern worldwide. This study aims to determine the prevalence of Enterobacterales producing beta-lactamase (TEM, SHV, OXA) or extended-spectrum beta-lactamases (ESBL), as well as plasmid-mediated resistance to quinolones (PMQR) (qnrA, qnrB, qnrS) in companion animals from the northeast region of Romania. A total of 124 faecal samples were collected aseptically from healthy dogs attending the veterinary practice for vaccination and cultivated on Brilliance ESBL medium (Oxoid, UK). The ESBL production testing was performed using the combination disc test. The identification of Enterobacterales strains was achieved using molecular identification and based on biochemical tests. Antimicrobial susceptibility testing was performed using the disk diffusion method. Identification of genes encoding for beta-lactamase enzymes and genes encoding plasmid-mediated resistance to quinolones was performed by PCR according to the protocols previously described. After ESBL screening, 31 (31/124; 25%) extended-spectrum cephalosporin (ESC)-resistant Enterobacterales were obtained, and 67.74% (21/31) of them were confirmed as ESBL-producers. Regarding the Enterobacterales species, 27 (27/31; 87.1%) were Escherichia coli and 4 (4/31; 12.9%) strains were Klebsiella pneumoniae. Among the ESBL-producing isolates, the blaCTX-M-1 gene group was predominant (58.82%), followed by the blaCTX-M-9 group (41.18%). The blaTEM, blaSHV and blaOXA gene groups were identified in 54.83%, 29.03% and 3.22% of the analysed strains, respectively. The prevalence of PMQR genes was 22.58% and consisted only of qnrS (19.35%) and qnrA (3.22%) genes. The prevalence of ESBL strains related to the total number of analysed samples was 16.93% (21/124). The findings show a significant prevalence of ESBLs and PMQR genes in Enterobacterales strains isolated from the faeces of healthy dogs, implying that pets may pose a risk of transmitting ESBL strains to other animals or owners.

Transfer and accumulation of antibiotic resistance genes and bacterial pathogens in the mice gut due to consumption of organic foods

Over the last few decades, organic food demand has grown largely because of increasing personal health concerns. Organic farming introduces antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) into foods. However, potential effects of organic foods on the gut microbiome and ARGs have been overlooked. Using high-throughput quantitative PCR and 16S rRNA high-throughput sequencing technology, we examined 132 ARGs from major classes, eight transposase genes, universal class I integron-integrase gene (intI), clinical class I integron-integrase gene (cintI), and the bacterial community in mouse gut after 8 weeks with an either organic or inorganic lettuce and wheat diet. A total of 8 types of major ARGs and 10 mobile genetic elements (MGEs) were detected in mice gut, including tetracycline, multidrug, sulfonamide, aminoglycoside, beta-lactamase, chloramphenicol, MLSB and vancomycin resistance genes. We found that abundance and diversity of ARGs, mobile gene elements, and potential ARB in the gut increased with time after consumption of organic foods, whereas no significant changes were observed in inorganic treated groups. Moreover, MGEs, including IS613, Tp614 and tnpA_03 were found to play an important role in regulating ARG profiles in the gut microbiome following consumption of organic foods. Importantly, feeding organic food increased the relative abundance of the potentially antibiotic-resistant pathogens, Bacteroides and Streptococcus. Our results confirm that there is an increasing risk of ARGs and ARB in the gut microbiome, which highlights the importance of organic food industries taking into account the potential accumulation and transmission of ARGs as a risk factor.

Molecular detection of Shiga toxin and extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates from sheep and goats.

The Shiga toxin (Stx)-producing Escherichia coli (STEC) have become important global public health concerns. This study investigated the prevalence, antimicrobial resistance profile, and extended-spectrum beta-lactamase-producing E. coli in sheep and goat faeces. A total of 53 E. coli isolates were confirmed by PCR targeting the uidA [β-D glucuronidase] gene. The Shiga toxin genes stx1 and stx2, as well as bfpA, vir, eaeA, lt and aafII virulence genes, were detected in this study. Of the 53 isolates confirmed to be STEC, 100% were positive for stx2 and 47.2% for stx1. Three isolates possessed a combination of stx1 + stx2 + eaeA, while four isolates harboured stx1 + stx2 + vir virulence genes. The isolates displayed phenotypic antimicrobial resistance against erythromycin (66.04%), colistin sulphate (43.4%), chloramphenicol (9.4%) and ciprofloxacin (1.9%). A total of 28.8% of the strains were phenotypically considered ESBL producers and contained the beta-lactamase blaCTX-M-9 and blaCTX-M-25 gene groups. A larger proportion of the E. coli strains (86.8%) contained the antibiotic sulphonamide resistant (sulII) gene, while 62.3%, 62.3%, 52.8%, 43.4%, 41.5%, 20.8%, 18.9%, 11.3%, 11.3%, 9.4%, 9.4% and 5.7% possessed mcr-4, floR, mcr-1, tet(A), sulI, tet(O), tet(W), parC, mcr-2, ampC 5, qnrS and ermB genes, respectively. Thirteen isolates of the ESBL-producing E. coli were considered multi-drug resistant (MDR). One Shiga toxin (stx2) and two beta-lactamase genes (blaCTX-M-9 and blaCTX-M-25 groups) were present in 16 isolates. In conclusion, the E. coli isolates from the small stock in this study contained a large array of high antibiotic resistance and virulence profiles. Our findings highlight the importance of sheep and goats as sources of virulence genes and MDR E. coli. From a public health and veterinary medicine perspective, the characterization of ESBL producers originating from small livestock (sheep and goats) is crucial due to their close contact with humans.

Municipal wastewater treatment plant showing a potential reservoir for clinically relevant MDR bacterial strains co-occurrence of ESBL genes and integron-integrase genes

Municipal wastewater treatment plants (MWWTPs) are a milieu for co-occurrence of multiple antibiotic resistance genes (ARGs). This facilitates mixing and genetic exchange; and promotes dissemination of multidrug resistance (MDR) to wastewater bacterial communities which is hazardous for the effluent receiving environment. This study investigated the co-occurrence of extended-spectrum beta-lactamase (ESBL) genes (blaTEM, blaCTX-M, blaSHV, blaOXA), and integron-integrase genes (intI1, intI2, intI3) in MDR bacteria isolated from the Bharwara MWWTP in Lucknow, India. Thirty-one MDR bacterial colonies resistant to three or more antibiotics were isolated from three treatment stages of this MWWTP. Six of these: Staphylococcus aureus, Serratia marcescens, Salmonella enterica, Shigella sonnei, Escherichia coli, and Bacillus sp. Had co-occurrence of ESBL and integron-integrase genes. These six isolates were examined for the occurrence of MDR efflux genes (qacA, acrB) and ARGs (aac(3)-1, qnrA1, tetA, vanA) and tested for resistance against 12 different antibiotics. The highest resistance was against penicillin-G (100%) and lowest for chloramphenicol (16.66%). Bacillus sp. Isolate BWKRC6 had the highest co-occurrence of antibiotic resistance-determining genes and was resistant to all the 12 antibiotics tested. The co-occurrence of ESBL, integron-integrase, antibiotic resistance-determining and MDR efflux genes in bacteria isolated from the Bharwara MWWTP indicates that the wastewaters of this treatment plant may have become a hotspot for MDR bacteria and may present human and environmental health hazards. Therefore, there is need for a rapid action to limit the spread of this threat. Public regulatory authorities must urgently implement measures to prevent MWWTPs becoming reservoirs for evolution of antibiotic resistance genes and development of antibiotic resistance.

Molecular Detection of Beta Lactamase Genes from Salmonella Typhimurium Isolated from Poultry Droppings in Nyanya, Abuja

Salmonella infections are one of the major global public health problems. During the last decade, antibiotic resistance and multiresistance of S. typhimurium. This study aimed at molecular detection of Beta Lactamase genes from S. typhimurium Isolated from poultry droppings in Nyanya, Abuja. Fecal droppings samples were collected from two different Poultry farms using sterile screw tubes. S. Typhimurium was isolated and identified using standard microbiology methods. Antimicrobial susceptibility was carried out using Clinical and Laboratory Standards Institute method. Out of 180 samples collected the overall occurrence of S. typhimurium was 20 (11.1%). S. typhimurium isolated were higher in Farm A 9(10.0%) than in farm B 11(12.2%). The S. typhimurium isolates from Farm A were highly resistant to Ampicillin with 77.7% but less resistant to gentamicin with 33.3%, ofloxacin, ciprofloxacin and Sulphamethoxazole/trimethoprim with 44.4%. Isolates from Farm B were highly resistant to Ampicillin and Sulphamethoxazole/trimethoprim 81.8%, but less resistant to Cefuroxime 36.3%. the most common phenotypic resistant observed were C-CIP-CAZ-CRO-AUG- OFL -SXT- CEF (22.2%) from Farm A. CIP-CRO-CAZ- AMP -AUG- OFL -SXT- CEF (18.1%) from Farm B, The commonest MAR index observed in this study was 0.8(33.3%) in farm A and (36.3%) in farm B. The order of occurrences of the 4 Beta Lactamase genes detected was: blaCTX-M-9 and blaSHV (75.0%) > bla CMY2 and Bla CTX-M (50.0%). The resistance observed in this study indicates it will be difficult to use commonly use antibiotics to treat infection caused by S. typhimurium isolated in the study location

Genetic characterization of virulence and extended spectrum β-lactamase producing genes of Klebsiella pneumoniae isolated from bovine milk.

Mastitis associated Klebsiella pneumoniae species were isolated from bovine milk to characterize virulence genes (wabG and kfuBC) and extended spectrum β-lactamase (ESBL) genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-9, blaTEM, blaSHV and blaOXA). A total number of 325 bovine milk samples (195 raw and 130 mastitic milk specimens) collected from Banaskantha, a milk-shed district of Gujarat, India, were included in the study. A total number of 27 K. pneumoniae isolates were recovered, consisting of 17 (62.96%) isolates from raw milk and 10 (37.03%) isolates from mastitic milk samples, giving an overall prevalence of 8.31%. Antibiotic sensitivity patterns revealed that 20 out of 27 isolates were found to be multi-drug resistant. Based on combination disc diffusion test and HiCrome ESBL agar method, 20 (74.07%) and 25 (92.59%) isolates were detected as ESBL producers, respectively. Among virulence genes studied, presence of wabG (25/27; 92.59%) was higher than kfuBC (5/27; 18.51%). Beta-lactamase genes viz., blaSHV, blaTEM and blaCTX-M-1 were detected in 23/27 (85.18%), 3/27 (11.11%) and 2/27 (7.40%) of isolates, respectively; while, none of the isolates was found to be positive for blaCTX-M-9 and blaOXA-1 genes. Outcome of the study provided an insight into virulence genes and ESBL producing K. pneumoniae isolated from bovine milk samples in India.

Frequency of Fecal Carriage of ESBL Resistance Genes in Multidrug-Resistant Pseudomonas aeruginosa Isolates from Cancer Patients at Laquintinie Hospital, Douala, Littoral Region, Cameroon.

Background. Opportunistic infections are the second cause of death among cancer patients. This study aimed at determining the antimicrobial profile and the prevalence of extended-spectrum beta-lactamase (ESBL)-gene carriage of Pseudomonas aeruginosa isolates among cancer patients at the Douala Laquintinie Hospital, Littoral Region of Cameroon. Between October 2021 and March 2023, 507 study participants were recruited among whom 307 (60.55%) were cancer patients, compared to 200 (39.45%) noncancer patients. Fifty-eight P. aeruginosa isolates were isolated from fecal samples of forty-five cancer patients and thirteen noncancer patients using Cetrimide agar. The antimicrobial resistance profile of the isolates was determined using the Kirby-Bauer disk diffusion method. The polymerase chain reaction was used to detect the presence of extended-spectrum beta-lactamase genes among P. aeruginosa isolates. P. aeruginosa showed significant resistance rates in cancer patients compared to noncancer patients to imipenem, cefotaxime, and ceftazidime, piperacillin-tazobactam, ticarcillin-clavulanic acid, and ciprofloxacin. The multidrug resistance (MDR) rate was significantly (p < 0.05) higher in cancer patients than in noncancer patients. The frequency of beta-lactamase genes in the 58 ESBL-producing P. aeruginosa isolates was determined as 72.41% for bla TEM, 37.93% for bla OXA, 74.14% for blaCTX-M, and 44.83% for bla SHV genes. The study revealed an alarmingly high prevalence of fecal carriage of ESBL-producing P. aeruginosa with a high rate of MDR among cancer patients. It indicates that regular monitoring and surveillance of ESBL-producing P. aeruginosa among cancer patients are needed to improve the management of patients.

Pathogenic Detection by Metagenomic Next-generation Sequencing in Skin and Soft Tissue Infection.

Skin and soft tissue infections (SSTIs) can be life-threatening, but the conventional bacterial cultures have low sensitivity and are time-consuming. Metagenomic next-generation sequencing (mNGS) is widely used as a diagnostic tool for detecting pathogens from infection sites. However, the use of mNGS for pathogen detection in SSTIs and related research is still relatively limited. From January 2020 to October 2021, 19 SSTI samples from 16 patients were collected in a single center (Taichung Veterans General Hospital, Taichung, Taiwan). The clinical samples were simultaneously subjected to mNGS and conventional bacterial culture methods to detect pathogens. Clinical characteristics were prospectively collected through electronic chart review. The microbiological findings from conventional bacterial culture and mNGS were analyzed and compared. The mNGS method detected a higher proportion of multiple pathogens in SSTIs compared to conventional bacterial culture methods. Pseudomonas spp. was among the most commonly identified Gram-negative bacilli using mNGS. Additionally, the mNGS method identified several rare pathogens in patients with SSTIs, including Granulicatella adiacens, Bacillus thuringiensis, and Bacteroides fragilis. Antimicrobial resistance genes were detected in 10 samples (52.6%) using the mNGS method, including genes for extended-spectrum beta-lactamase, Ambler class C β-lactamases, and carbapenemase. mNGS not only plays an important role in the detection of pathogens in soft tissue infections, but also informs clinical professionals about the presence of additional microbes that may be important for treatment decisions. Further studies comparing conventional pathogen culture with the mNGS method in SSTIs are required.

Investigation of antibiotic resistance, virulence genes, and biofilm formation of Escherichia coli isolated from sheep feces in Shiraz industrial slaughterhouse, South of Iran.

With the increase in human population, the consumption of livestock products such as sheep meat has also increased. Sheep are the reservoir and shedder of Escherichia coli that can be transmitted to humans. Aims: Characterization of fecal E. coli isolated from sheep in slaughterhouse. Stool specimens were collected from 30 apparently healthy sheep from different flocks in Shiraz industrial slaughterhouse. The resistance of E. coli isolates against 10 antibiotics was determined by disk diffusion method. The presence of three major extended spectrum beta-lactamase (ESBL) genes and five tetracycline resistance genes as well as seven virulence genes were investigated by polymerase chain reaction (PCR) technique. Using the microtiter plate method, the biofilm formation ability of E. coli isolates was investigated. The highest frequency of resistance was to amoxicillin (100%) followed by tetracycline (25%). All E. coli isolates were susceptible to gentamicin and nitrofurantoin, and only one isolate was resistant to the tested third-generation cephalosporins. Multidrug resistance phenotype was observed in 16.7% of the isolates. bla TEM (25%) was the most prevalent ESBL gene and tetA (62.5%) was the most prevalent tetracycline resistance gene in the isolates. crl, csgA, fimH, and bcsA genes were present in all isolates, and the prevalence of papC and afa genes was 95.8% and 83.3%, respectively. In total, 62.5% of the isolates were biofilm producers. According to the concept of One Health, the presence of virulent antibiotic-resistant biofilm producing strains of E. coli in sheep is a risk to public health.

Antibiotypes and genetic characteristics of fluoroquinolone- and beta-lactam-resistant Escherichia coli isolated from food-producing animals

Background and Aim: Farm animals, including cattle, have been implicated as antimicrobial-resistant bacterial pathogen reservoirs. This study aimed to determine the antimicrobial resistance profiles and genetic characteristics of cattle colonized by fluoroquinolone-resistant and extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in Ebonyi state, Nigeria. Materials and Methods: We randomly collected 100 fresh fecal samples from 100 cattle in major abattoirs and analyzed the samples using standard microbiological methods. Isolates were further characterized by polymerase chain reaction (PCR) using 16S rRNA sequence primers. Phenotypic detection of ESBL production was performed using the double disk synergy test. Antimicrobial susceptibility profiles of ESBL-producing Escherichia coli were determined using the disk diffusion method, whereas molecular characterization of ESBL- and fluoroquinolone-resistant genes was performed by PCR using specific primers. Results: A total of 20 (20%) ESBL-producing E. coli were isolated from 100 animal fecal samples. Isolates were generally multidrug-resistant (MDR) with a resistance rate of 100% to 45% to trimethoprim-sulfamethoxazole, tetracycline, amoxicillin, cephalosporins, and ciprofloxacin. The average multiple antibiotic resistance index values of the isolates ranged from 0.5 to 0.8. BlaTEM (75%), followed by blaCTX-M (20%) and blaSHV (5.0%) was the most predominant ESBL gene among the isolates. The Aac-lb-6-cr fluoroquinolone-resistant gene was harbored by 90% of the isolates, whereas Qnr was absent. Conclusion: This study showed a high frequency of MDR ESBL-producing E. coli harboring ESBL and fluoroquinolone-resistant genes in fecal samples of cattle with serious public health consequences if not adequately addressed. Keywords: Escherichia coli, extended-spectrum beta-lactamase genes, fluoroquinolone resistance genes, cattle, multidrug-resistance.

Evaluation of Multidrug Resistance and TEM and SHV Broad-Spectrum Beta-Lactamase Genes in Escherichia coli Obtained from Patients with Urinary Tract Infections in Alborz Province, Iran

Evaluation of Multidrug Resistance and TEM and SHV Broad-Spectrum Beta-Lactamase Genes in Escherichia coli Obtained from Patients with Urinary Tract Infections in Alborz Province, Iran

Characterization of extended-spectrum beta-lactamase and carbapenemase genes in bacteria from environment in Burkina Faso.

This study aimed to characterize extended-spectrum beta-lactamase (ESBL) and carbapenemase genes in bacteria from the environment in Bobo-Dioulasso, Burkina Faso. This study was conducted from January 18 to December 31, 2019. Environmental samples were collected from the effluents of Souro Sanou University Hospital Center and the wastewater treatment plant at Bobo-Dioulasso. MacConkey agar media supplemented with 4 µg/mL cefotaxime was used for bacterial growth, and identification of bacteria was performed using API 20E system (BioMerieux SA, Lyon, France). Antibiotic susceptibility testing, synergy test, carbapenem inactivation method and molecular characterization were performed. A total of 180 bacterial isolates were identified from the different sites with a predominance of Klebsiella oxytoca and Klebsiella pneumoniae (27.5%). All 180 bacterial isolates were ESBL producers and 18 (10.0%) of them produced carbapenemases. Out of the 180 bacterial isolates, DNAs of 98.9% (178/180) bacterial isolates were extracted and tested through polymerase chain reaction (PCR) for characterization of resistant genes. The study showed that 89.8% (160/178) carried the bla-CTX-M genes including 54.4 (87/160) from hospital effluents and 45.6 (73/160) from the wastewater treatment plant. Regarding the carriage of carbapenemase genes, 7.9 (14/178) blaNDM-1 was found in all the sites including 71.4% (10/14) from hospital effluents and 28.6 (4/14) from the wastewater treatment plant. blaOXA-48-like was only found in bacteria from hospital effluents and represented 2.2% (4/178). This study highlights the need to build hospital effluent treatment plants to reduce the load of resistant bacteria before discharging the effluents into the urban wastewater system.

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&amp;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.

Visualizing and quantifying structural diversity around mobile resistance genes.

Understanding the evolution of mobile genes is important for understanding the spread of antimicrobial resistance (AMR). Many clinically important AMR genes have been mobilized by mobile genetic elements (MGEs) on the kilobase scale, such as integrons and transposons, which can integrate into both chromosomes and plasmids and lead to rapid spread of the gene through bacterial populations. Looking at the flanking regions of these mobile genes in diverse genomes can highlight common structures and reveal patterns of MGE spread. However, historically this has been a largely descriptive process, relying on gene annotation and expert knowledge. Here we describe a general method to visualize and quantify the structural diversity around genes using pangraph to find blocks of homologous sequence. We apply this method to a set of 12 clinically important beta-lactamase genes and provide interactive visualizations of their flanking regions at https://liampshaw.github.io/flanking-regions. We show that nucleotide-level variation in the mobile gene itself generally correlates with increased structural diversity in its flanking regions, demonstrating a relationship between rates of mutational evolution and rates of structural evolution, and find a bias for greater structural diversity upstream. Our framework is a starting point to investigate general rules that apply to the horizontal spread of new genes through bacterial populations.

Evaluation and clinical practice of pathogens and antimicrobial resistance genes of BioFire FilmArray Pneumonia panel in lower respiratory tract infections

BackgroundExisting panels for lower respiratory tract infections (LRTIs) are slow and lack quantification of important pathogens and antimicrobial resistance, which are not solely responsible for their complex etiology and antibiotic resistance. BioFire FilmArray Pneumonia (PN) panels may provide rapid information on their etiology.MethodsThe bronchoalveolar lavage fluid of 187 patients with LRTIs was simultaneously analyzed using a PN panel and cultivation, and the impact of the PN panel on clinical practice was assessed. The primary endpoint was to compare the consistency between the PN panel and conventional microbiology in terms of etiology and drug resistance, as well as to explore the clinical significance of the PN panel. The secondary endpoint was pathogen detection using the PN panel in patients with community-acquired pneumonia (CAP) or hospital-acquired pneumonia (HAP).ResultsFifty-seven patients with HAP and 130 with CAP were included. The most common pathogens of HAP were Acinetobacter baumannii and Klebsiella pneumoniae, with the most prevalent antimicrobial resistance (AMR) genes being CTX-M and KPC. For CAP, the most common pathogens were Haemophilus influenzae and Staphylococcus aureus, with the most frequent AMR genes being CTX-M and VIM. Compared with routine bacterial culture, the PN panel demonstrated an 85% combined positive percent agreement (PPA) and 92% negative percent agreement (NPA) for the qualitative identification of 13 bacterial targets. PN detection of bacteria with higher levels of semi-quantitative bacteria was associated with more positive bacterial cultures. Positive concordance between phenotypic resistance and the presence of corresponding AMR determinants was 85%, with 90% positive agreement between CTX-M-type extended-spectrum beta-lactamase gene type and phenotype and 100% agreement for mecA/C and MREJ. The clinical benefit of the PN panel increased by 25.97% compared with traditional cultural tests.ConclusionThe bacterial pathogens and AMR identified by the PN panel were in good agreement with conventional cultivation, and the clinical benefit of the PN panel increased by 25.97% compared with traditional detection. Therefore, the PN panel is recommended for patients with CAP or HAP who require prompt pathogen diagnosis and resistance identification.

Characterization of VIM, VEB and CTX-M Beta-lactamase Gene in Escherichia coli and Pseudomonas aeruginosa Isolated from Urine Samples of Patients Visiting a Tertiary Hospital in Abakaliki

The spread and convergence of multiple beta-lactamase genes across distinct resistant bacterial populations from various hosts and settings demonstrates increased risk of morbidity and mortality in humans. This study was undertaken to characterize blaVIM, blaVEB and blaCTX-M beta-lactamase gene in Escherichia coli and P. aeruginosa isolates from patients visiting a tertiary hospital in Abakaliki. A total of three hundred (300) urine samples were collected from patients and were subjected to bacteriological examination using culture, Gram staining and biochemical technique, for routine microbiological identification and further confirmed using the VITEK-2 Automated System (Biomerieux, France). Antimicrobial susceptibility studies were determined using the Kirby–Bauer disk diffusion method. All isolate were further screen for various beta-lactamase resistant gene by PCR using specific primer. Of the 300 urine samples collected, prevalence rate of 187 (62.3%) and 91 (30.3 %) E. coli and P. aeruginosa were recorded. The isolates exhibited 50.0-100% percentage of resistance to Amoxycillin-Clavulanic acid, Azetronam, Cefoxitin, Ceftriaxone and Piperacillin/tazobactam. The proportion of beta-lactamase gene in E. coli were as follows (VEB 143/76.5 %; CTX-M 175/93.5 %; VIM 77/41.2 %) while beta-lactamase gene in P. aeruginosa were as follows (VEB 91/100 %; CTX-M 63/69.2%; VIM 48/52.7 %). The presence of these gene in our study indicates the possibility of therapeutic failure, serious consequences for infection control and increased risk of morbidity and mortality in patients. Hence, continuous effort in hospital surveillance, infection control, and clinical audits must be conducted to fight against the rapid development and spread of antibiotic-resistant bacteria pathogens. Keywords: Beta-lactamase, Escherichia coli, Pseudomonas aeruginosa, VIM, VEB, CTX-M