Bla Gene Research Articles

Phenotypic and Molecular Detection of Extended Spectrum Beta-Lactamases Producing-Bacteria Isolated From Pregnant Women With Genital Tract Infection in Duhok Governorate

Beta-lactamase producing bacteria have a worldwide distribution with a high degree of prevalence in both community and hospital. Furthermore, multidrug resistant (MDR) and extended spectrum β-lactamases (ESBL) producing bacterial isolates from women patients may limit treatment options available. This study was designed to determine the frequency of bacterial isolates associated with genital tract infection in pregnant women and their antimicrobial resistance profile and to assess the prevalence of extended spectrum β-lactamases producing bacteria. Demonstrating the β-lactamase genes (blaTEM, blaSHV and blaCTX-M) by using polymerase chain reaction (PCR) assay with specific primers, was carried out on patients who were admitted to Maternity and Obstetric Hospital in Duhok city from November 2018 to October 2019. A total of 100 high vaginal swabs were collected from pregnant women patients between the ages 18-45 years. All clinical samples were cultured and standard microbiological methods were used to identify bacterial isolates, then confirmed by Vitek®2 compact automated system. All gram negative bacterial isolates were studied phenotypically and genotypically for extended spectrum β-lactamases-production. Out of 100 vaginal swabs, 88% confirmed positive culture; 90.9% of which were bacterial isolates. From the total bacterial isolates, 38.8% were gram negative bacteria, with a predominant 54.8% Klebsiella pneumoniae followed by Escherichia coli 35.5%. 54.8% of the isolates were characterized as multidrug resistance isolates, 29% isolates were extensive drug resistance, and no pan drug resistance were detected. Among these, the commonest extended spectrum β-lactamases producing isolates were Escherichia coli 81.8% followed by, Klebsiella pneumoniae 58.8%. Extended spectrum β-lactamases-producing isolates have showed significantly higher resistance than non- extended spectrum β-lactamases producing isolates to third and fourth generation cephalosporins. CTX-M was the most common β-lactamase gene 73.7% among extended spectrum β-lactamase producing strains, followed by blaSHV, 57.9% and blaTEM 52.6%, 21.1% had combination of all bla genes, 15.8% had CTX-M only and combination of blaCTX-M with blaSHV and blaTEM. 10.5% among extended spectrum β-lactamases producing isolates carried SHV type only and in combination with TEM type while TEM gene were observed in 5.3%. We concluded that the drug resistant isolates were common, worryingly high and it may limit treatment options available. In this study a high level of the blaCTX-M gene was demonstrated among extended spectrum β-lactamases producing isolates.

Plasmid-Encoded AmpC and Extended-Spectrum Beta-Lactamases in Multidrug-Resistant Escherichia coli Isolated from Piglets in Portugal.

Considering the concept of "One Health," the aim of this study was to determine susceptibility profiles of Escherichia coli in piglets' intestinal microbiota from different farms in Portugal. Beyond antimicrobial susceptibility, the occurrence of multiple antibiotic resistance and detection of phenotypic/genotypic extended-spectrum beta-lactamases (ESBLs) and plasmid mediated AmpC beta-lactamases (pAmpC) were done. From 10 different pig farms, 340 E. coli isolates were obtained from 75 feces samples. Susceptibility to amoxicillin-clavulanic acid (AMC), piperacillin (PIP), cefoxitin (FOX), ceftazidime (CAZ), cefepime (FEP), aztreonam (AZT), imipenem (IP), amikacin (AK), ciprofloxacin (CIP), and trimethoprim-sulfamethoxazole (SXT) was determined. Five-gene panel for amplification of bla genes was used for ESBL (TEM, SHV, CTX-M) and pAmpC (CMY-2, ACC). Among E. coli isolates, 209 were distributed in three resistance profiles: 57.7% MDR, 3.5% extensively drug-resistance (XDR) (resistant to CIP, SXT, and beta-lactams, except IP, with variability to AK) and 0.3% pandrug-resistance (PDR) (resistant to all antibiotics used). pAmpC and/or ESBLs genes were presented in 65% of the isolates. Presence of different associations of bla genes in the same isolate was the most observed (31%), and the most common were an ESBL (TEM) and a pAmpC (CMY-2). Presence of three or four bla genes in various associations were detected. These isolates were very resistant, especially those with four genes, which were resistant to beta-lactams (except IP), CIP, and SXT. This study showed a surprisingly high rate of MDR E. coli isolated in Portuguese piglets, with enzymes that impair activity of the most used antibiotics in human therapeutic.

Molecular characterization of carbapenem-resistant Acinetobacter baumannii clinical isolates from Egyptian patients.

Acinetobacter baumannii (A. baumannii) represents a global threat owing to its ability to resist most of the currently available antimicrobial agents. Moreover, emergence of carbapenem resistant A. baumannii (CR-AB) isolates limits the available treatment options. Enzymatic degradation by variety of ß-lactamases, have been identified as the most common mechanism of carbapenem resistance in A. baumannii. The alarming increase in the prevalence of CR-AB necessitates continuous screening and molecular characterization to appreciate the problem. The present study was performed to assess the prevalence and characterize carbapenemases among 206 CR-AB isolated from various clinical specimens collected from different intensive care units at Kasr Al-Aini Hospital. All isolates were confirmed to be A. baumannii by detection of the blaOXA-51-like gene. Molecular screening of 13 common Ambler class bla carbapenemases genes in addition to insertion sequence (IS-1) upstream OXA-23 were performed by using four sets of multiplex PCR, followed by identification using gene sequencing technology. Among the investigated genes, the prevalence of blaOXA-23, and blaOXA-58 were 77.7%, and 1.9%, respectively. The ISAba1 was detected in 10% of the blaOXA-23 positive isolates. The prevalence of metallo-β-lactamases (MBLs) studied; blaNDM-1, blaSPM, blaVIM, blaSIM-1 were 11.7%, 6.3%, 0.5%, and 0.5% respectively. One of class A; bla KPC was detected in 10.7% of the investigated isolates. blaOXA-24/40, blaIMP, blaGES, blaVEB and blaGIM were not detected in any of the studied isolates. Moreover, 18.4% of the isolates have shown to harbor two or more of the screened bla genes. We concluded that the most prevalent type of ß-lactamases genes among CR-AB isolates collected from Egyptian patients were blaOXA-23 followed by blaNDM-1 and blaKPC.

Antimicrobial resistance profiles and genetic basis of resistance among non-fastidious Gram-negative bacteria recovered from ready-to-eat foods in Kibera informal housing in Nairobi, Kenya

ObjectiveThis cross-sectional study conducted in Kibera, Kenya, sought to gain insights on relative microbial contamination levels of popular unprocessed food types, determine antimicrobial resistance (AMR) burden and the carriage of integrons that are essential elements for spreading antimicrobial resistance genes (ARG). Foods analysed consisted of cooked vegetables (kale, cabbage, and nightshades), boiled cereal foods (beans, rice, and Githeri, which is a mixture of beans and maize), meat, Omena fish (fried silver cyprinids), and Ugali (a product of simmered maize flour in boiled water).ResultsThe analysis detected contamination levels exceeding 2×104 c.f.u. ml−1 in 106 (38 %) of the 281 ready-to-eat foods analysed. The majority of food types had microbial contaminations of between 4.0×104 and 2.3×106 c.f.u. ml−1. Kale was the most contaminated with a mean of 2.3×106 c.f.u. ml−1, while Omena was the least contaminated with 4.0×104 c.f.u. ml−1. Foods sold close to open sewage and refuse sites were more contaminated than those sold in relatively ‘cleaner’ settings (P <0.0001, O.R 0.1162, C.I 0.1162–0.120). A total of 405 bacterial isolates were recovered and included; Klebsiella spp 116 (29 %), Escherichia coli 104 (26 %), Enterobacter agglomerans 88 (22 %), Proteus mirabilis 30 (7 %), Salmonella spp 28 (7 %), Citrobacter freundii 27 (7 %) and Serratia marcescens 12 (3 %). Imipenem (IPM, 100 %) was the most effective antimicrobial agent, followed by cefepime (98 %). Ampicillin (AMP, 33 %), trimethoprim (TMP, 27 %), and sulfamethoxazole (SMX, 23 %) on the other hand, were the least effective antimicrobials. The analysis also found ten isolates (2 %) that had co-resistance to third-generation cephalosporins, fluoroquinolone (CIP), quinolones (NAL) and aminoglycosides (GEN); hereby we refer to this phenotype as the βFQA. The prevalence of multidrug-resistant (MDR) strains was 23 % (93), while that of extended-spectrum β-lactamases (ESBL) producing strains was 4 % (17). The bla TEM was the most prevalent (55 %) β-lactamase (bla) gene among the screened 93 MDR-strains. Carriage of class one integrons (intI1) was more common (23 %) than intl2 (3 %) among these MDR-strains. Bacterial diversity analysis using the GTG5-PCR found no significant clusters for analysed E. coli and K. pneumoniae, suggesting recovered isolates were genetically diverse and not due to non-clonal expansion. The findings of this study are an indication that contaminated foods can be a reservoir for enteric pathogens and a source of AMR strains.

Carbapenem resistance in Acinetobacter baumannii clinical isolates from northwest Iran: high prevalence of OXA genes in sync.

Background and Objectives:Carbapenem treatment for Acinetobacter baumannii infections presently faces threats owing to the production of several types of carbapenemase enzymes, prevalence of which varies among different countries. We explored the current trend of antibiotic resistance in A. baumannii clinical isolates from North West Iran, sought the mechanism of carbapenem resistance and addressed the sequence type groups in carbapenem resistant A. baumannii (CRAB).Materials and Methods:A. baumannii (n=112) isolates were recovered from various clinical specimens of patients admitted in internal, surgery, burn, infectious diseases and various ICUs wards. Genetically confirmed A. baumannii isolates were screened for carbapenem resistance by the Kirby-Bauer and E-test and the presence of bla MBL, bla OXA-like, ISAba1 genes by PCR. Sequence groups were identified by multiplex PCR.Results:Multidrug-resistance (MDR) was a characteristic feature of all A. baumannii isolates. Frequency of oxacillinase genes in combination including bla OXA-51-like/bla OXA-23-like, bla OXA-51-like/blaOXA-24/40-likeand bla OXA-51-like/bla OXA-23-like/bla OXA-24/40-like was 82.1%, 36.6% and 25.8% respectively. Blending of oxacillinase and MBL genes was evident in eight bla OXA-23-like positive and 7 bla OXA-24-like positive isolates thereby depicting synchronous etiology of carbapenem resistance. Amongst CRAB isolates, 97.3% contained ISAba1 element and 50.9% belonged to the European clone II.Conclusion:Synchronicity among bla OXA-like with bla MBL and ISAba1 gene was a hallmark of this investigation. Though origin or route of transmission was not elucidated in this study but co-existence among OXA and MBL producing genes is a therapeutic concern demanding strict surveillance strategies and control programs to halt the dissemination of these isolates in the hospital setting.

Prevalence of extended-spectrum beta-lactamase-producing Escherichia coli strains in dairy farm wastewater in Chiang Mai

We investigated the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains in dairy farm wastewater in Chiang Mai, Thailand. We analyzed wastewater samples collected from 150 dairy farms and found that 88.7% of the farms (n = 133) were positive for ESBL-producing E. coli. Multiplex polymerase chain reaction (PCR) amplification was performed to characterize the presence of bla CTX-M, bla TEM, and blaSHV in ESBL-producing isolates. blaCTX-M was found in all isolates (n = 133), followed by blaTEM (80/133, 60.2%), whereas blaSHV was not detected in any isolate. blaCTX-M and blaTEM were present in 60.2% (80/133) of the isolates, and 39.8% (53/133) isolates carried bla CTX-M alone. Subgroup analysis showed that CTX-M-1 was the most prevalent subgroup among the isolates (129/133, 97.0%), followed by CTX-M-8 (2/133, 1.5%) and CTX-M-9 (2/133, 1.5%). The distribution of the phylogenetic groups was as follows: group A (100/133, 75.2%), followed by B1 (14/133, 10.5%), D (6/133, 4.5%), F (6/133, 4.5%), B2 (4/133, 3.0%), and E (3/133, 2.3%). Based on enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and dendrogram analysis, 24 isolates were classified into clades I (n = 21), II (n =1), and III (n =2). Minor genetic differences were found in all clade I isolates. Our data suggest that the circulating of ESBL-producing E. coli carried at least one bla gene strain distributed in dairy farm wastewater in Chiang Mai.

Molecular Detection of Extended Spectrum Beta-lactamase Resistance in Escherichia coli from Poultry Droppings in Karu, Nasarawa State, Nigeria

Aims: This study investigates and reports the production of extended spectrum beta-lactamase in Escherichia coli isolates in poultry droppings sourced from selected poultry farms in Karu, Nigeria&#x0D; Study Design: Cross sectional study&#x0D; Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between August 2019 and February 2020.&#x0D; Methodology: Escherichia coli was isolated from the samples using standard cultural and microbiological methods. Antibiotic susceptibility testing and minimum inhibitory concentrations were evaluated as described by the Clinical and Laboratory Standards Institute (CLSI). The detection of ESBL production in E. coli isolates was carried out using double disc synergy test. In addition, molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method.&#x0D; Results: All (100%) samples collected had E. coli. Antibiotic resistances in the isolates in decreasing order were as follows: ampicillin (96.7%), streptomycin (94.4%), sulphamethoxazole /trimethoprim (87.8%), amoxicillin/ clavulanic acid (61.1%), gentamicin (52.2%), ciprofloxacin (40.0%), ceftazidime (35.6%), cefotaxime (31.1%), imipenems (22.2%), cefoxitin (13.3%). The commonest antibiotic resistant phenotype was AMP-SXT-S-CTX-CN (8.8%). Multiple antibiotic resistance (MAR) was observed in 92.2% (83/90) of the isolates with the common MAR indices being 0.5 (26.5%), 0.6 (19.2%), 0.4 (13.2%) and 0.9 (10.8%). Fifty nine of the eighty beta-lactam resistant isolates (73.7%) were confirmed ESBL producers. 55 of the 59 ESBL positive isolates (93.2%) carried bla genes as follows: blaSHV (50/55, 90.9%), blaTEM (31/55, 56.3%) and blaCTX-M (46/55, 83.6%). Thirty six (65.5%) of the 55 isolates carried two bla genes (blaSHV and blaTEM, blaTEM and blaCTX-M, and blaCTX-M and blaSHV).&#x0D; Conclusion: The E. coli isolates showed lower resistances to cefoxitin, imipenem, cefotaxime, ceftazidime, and ciprofloxacin and most isolates were MAR, with resistance to 5 antibiotics being the most predominant. In addition, blaSHV gene was the most common ESBL gene detected in the confirmed ESBL-producing E. coli isolates.

Plasmids conferring resistance to extended-spectrum beta-lactamases including a rare IncN+IncR multireplicon carrying blaCTX-M-1 in Escherichia coli recovered from migrating barnacle geese ( Branta leucopsis).

Background: Increasing antimicrobial resistance (AMR) is a global threat and wild migratory birds may act as mediators of resistant bacteria across country borders. Our objective was to study extended-spectrum beta-lactamase (ESBL) and plasmid-encoded AmpC (pAmpC) producing Escherichia coli in barnacle geese using whole genome sequencing (WGS) and to identify plasmids harboring bla genes. Methods: Barnacle geese feces (n=200) were collected during fall 2017 and spring 2018 from an urban area in Helsinki, Finland. ESBL/AmpC-producing E. coli were recovered from nine samples (4.5%) and isolates were subjected to WGS on both short- and long-read sequencers, enabling hybrid assembly and determination of the genomic location of bla genes. Results: A rare multireplicon IncN+IncR was recovered from one isolate carrying bla CTX-M-1 in addition to aadA2b, lnu(F), and qnrS1. Moreover, rarely detected IncY plasmids in two isolates were found to harbor multiple resistance genes in addition to the human-associated bla CTX-M-15. Poultry-associated bla CMY-2 was identified from the widely distributed IncI1 and IncK plasmids from four different isolates. One isolate harbored an IncI1 plasmid with bla CTX-M-1 and flor. A chromosomal point mutation in the AmpC promoter was identified in one of the isolates. WGS analysis showed isolates carried multiple resistance and virulence genes and harbored multiple different plasmid replicons in addition to bla-carrying plasmids. Conclusions: Our findings suggest that wild migratory birds serve as a limited source of ESBL/AmpC-producing E. coli and may act as disseminators of the epidemic plasmid types IncI1 and IncK but also rarely detected plasmid types carrying multidrug resistance. Human and livestock-associated ESBL enzyme types were recovered from samples, suggesting a potential for interspecies transmission. WGS offers a thorough method for studying AMR from different sources and should be implemented more widely in the future for AMR surveillance and detection. Understanding plasmid epidemiology is vital for efforts to mitigate global AMR spread.

Spread of Multi-Antibiotic-Resistant Health-Care Pathogens in Hospitals to Treat COVID-19 Patients

Relevance. The COVID-19 pandemic has led to significant overloads in the work of health systems in many countries, a shortage of beds and staff, which contributes to a decrease in adherence to measures to prevent and control nosocomial infections, which can significantly worsen the course of viral pneumonia. Aim. To assess the possibility of the formation of hospital strains of multidrugresistant microorganisms in hospitals repurposed to provide medical care to patients with COVID-19. Materials and methods. The study included patients with severe and moderate forms of COVID-19 (ICD codes U07.1, U07.2), who were admitted to two large hospitals repurposed for the treatment of this infection. The data of microbiological studies of the biomaterial associated with the respiratory tract (sputum, bronchoalveolar lavage, tracheal aspirates) obtained from 1101 patients from May to January 2021 were analyzed using a combination of molecular genetic methods (RAPD-PCR, detection of integrons and the carbapenemase gene bla NDM.), and molecular typing of carbapenem-resistant strains of Klebsiella pneumoniae and Acinetobacter baumannii was carried out. Results. It was found that carbapenem resistant gram-negative bacteria predominate in the structure of the nosocomial microbiota of the respiratory tract of patients with COVID-19 in both hospitals. Based on molecular typing made the wide distribution of several genetic lines of integron-positive carbapenem resistant Klebsiella pneumoniae and Acinetobacter baumannii was detected. Conclusions. The COVID-19 pandemic has exacerbated the spread and circulation of bacteria with multiple antibiotic resistance in hospitals. This study has demonstrated the possibility of the formation of hospital strains of nosocomial infections in COVID-19 hospitals, which justifies the need to improve infection control measures in the context of a new coronavirus infection pandemic.

Identification of β-Lactamase-Encoding (bla) Genes in Phenotypically β-Lactam-Resistant Escherichia coli Isolated from Young Calves in Belgium.

The bla genes identification present in 94 phenotypically resistant Escherichia coli isolated from feces or intestinal contents of young calves with diarrhea or enteritis in Belgium was performed by microarrays (MA) and whole genome sequencing (WGS). According to their resistance phenotypes to 8 β-lactams at the disk diffusion assay these 94 E. coli produced a narrow-spectrum-β-lactamase (NSBL), an extended-spectrum-β-lactamase (ESBL) or a cephalosporinase (AmpC). All ESBL-encoding genes identified by MA and WGS belonged to the blaCTX-M family, with a majority to the blaCTX-M-1 subfamily. Two different genes encoding an AmpC, blaCMY-2, and blaDHA-1 were detected in isolates with an AmpC phenotype. The blaTEM-1 and the blaOXA-1 were detected alone in isolates with a NSBL phenotype or in combination with ESBL-/AmpC-encoding bla genes. Furthermore, the WGS identified mutations in the ampC gene promoter at nucleotides -42 (C>T) and/or -18 (G>A) that could not be identified by MA, in several isolates with an AmpC-like resistance phenotype. No carbapenemase-encoding gene was detected. To our knowledge this is the first survey on the identification of bla genes in E. coli isolated from young diarrheic or septicemic calves in Belgium.

Distribution of genetic elements associated with antibiotic resistance in treated and untreated animal husbandry waste and wastewater

Animal breeding for meat production based on swine, cattle, poultry, and aquaculture is an activity that generates several impacts on the environment, among them the spread of antibiotic resistance. There is a worldwide concern related to the massive use of antibiotics, which causes selective pressure on the microbial community, triggering bacteria that contain "antibiotic resistance genes." According to the survey here presented, antibiotic resistance-related genes such as tetracyclines (tet), erythromycin (erm), and sulfonamides (sul), as well as the genetic mobile element interferon (int), are the most reported genetic elements in qualitative and quantitative studies of swine, cattle, poultry, and aquaculture manure/wastewater. It has been observed that biological treatments based on waste composting and anaerobic digestion are effective in ARG removal, particularly for tet, bla, erm, and qnr (quinolone) genes. On the other hand, sul and intI genes were more persistent in such treatments. Tertiary treatments, such advanced oxidative processes, are suitable strategies to improve ARG reduction. In general temperature, hydraulic retention time, and penetration of sunlight are the main operational parameters for ARG reduction in treatments applied to animal waste, and therefore attention should be addressed to optimize their efficacy regarding ARG removal. Despite being reduced, the presence of ARG in treated effluents and in biosolids indicates that there is a potential risk of antibiotic resistance spread in nature, especially through the release of treated livestock waste into the environment.

Differential effects of chromosome and plasmid blaCTX-M-15 genes on antibiotic susceptibilities in extended-spectrum beta-lactamase-producing Escherichia coli isolates from patients with urinary tract infection.

To compare antibiotic susceptibilities between chromosomal and plasmid blaCTX-M-15 locations in urinary tract infection-causing extended-spectrum β-lactamases-producing Escherichia coli blaCTX-M-15 isolated in Indonesia. A total of 84 strains identified as extended-spectrum β-lactamases-producing E. coli were isolated from patients with urinary tract infection in Indonesia in 2015. Antimicrobial susceptibility tests were performed on these strains using 18 antibiotics, and extended-spectrum β-lactamase bla genes were detected by polymerase chain reaction. Gene localization of blaCTX-M-15 -positive strains was confirmed by Southern blot hybridization, and epidemiological typing was conducted using multilocus sequence typing. Of 54 strains harboring the blaCTX-M-15 gene, 27 showed localization on chromosome, 20 on plasmid, and seven on chromosome and plasmid. Most multilocus sequence typing sequence types of the 27 strains with chromosomal blaCTX-M-15 were ST405 (25.9%) and ST131 (22.2%) strains, whereas the 20 strains with plasmid-blaCTX-M-15 were mostly ST410 (55.0%). Extended-spectrum β-lactamases-producing E. coli blaCTX-M-15 with plasmid genes show significantly higher resistant rates against piperacillin-tazobactam but lower resistant rates against chloramphenicol compared to chromosomal strains in Indonesian patients with urinary tract infection. Mechanistic investigations will be necessary to advance our knowledge of antimicrobial resistance in urinary tract infection.

Treatment of pharmaceutical wastewater by ionizing radiation: Removal of antibiotics, antimicrobial resistance genes and antimicrobial activity

In present study, the treatment of real pharmaceutical wastewater from an erythromycin (ERY) production factory by gamma irradiation was investigated. Results showed that a variety of antimicrobial resistance genes (ARGs), involving MLSB, tet, bla, multidrug, sul, MGEs and van genes and plentiful 9 bacterial phyla were identified in the raw wastewater. In addition to ERY, sulfamethoxazole (SMX) and tetracycline (TC) were also identified with the concentration of 3 order of magnitude lower than ERY. Results showed that the abatement of ARGs and antibiotics was much higher than that of antimicrobial activity and COD. With the absorbed dose of 50 kGy, the removal percentage of ARGs, ERY, antimicrobial activity and COD was 96.5–99.8%, 90.0%, 47.8% and 10.3%, respectively. The culturable bacteria were abated fast and completely at 5.0 kGy during gamma irradiation. The genus Pseudomonas was predominant in raw wastewater (56.7%) and its relative abundance decreased after gamma irradiation, to 1.3% at 50 kGy. With addition of peroxymonosulfate (PMS, 50 mM), the antimicrobial activity disappeared completely and ERY removal reached as high as 99.2% at the lower absorbed dose of 25 kGy. Ionizing radiation-coupled technique is a potential option to treat pharmaceutical wastewater for reduction of antibiotics, ARGs and antimicrobial activity.

Draft Genome Sequences of Seven Extended-Spectrum β-Lactamase-Producing Escherichia coli Strains Isolated from New Zealand Waterways.

Draft genomes of seven extended-spectrum β-lactamase (ESBL)-producing Escherichia coli strains recovered from New Zealand waterways are described. The mean genome size was 5.1 Mb, with 4,724 coding sequences. All genomes contained the ESBL gene bla CTX-M, and one carried a plasmid-mediated AmpC gene, bla CMY-2 A multidrug-resistant genotype was detected in three isolates.

Characterization of antibiotic resistance profiles in Pseudomonas aeruginosa isolates from burn patients

ObjectiveTo investigate the prevalence of multidrug-resistant (MDR) Pseudomonas aeruginosa (PA) producing extended-spectrum beta-lactamases (ESBLs) and metallo-beta-lactamases (MBLs) in burn patients in Algeria. MethodsBetween April 2016 and October 2019, 47 non-redundant isolates of PA were collected from 47 burn patients admitted to the Department of Burns at the Military Hospital of Algiers in Algeria. Antibiotic susceptibility testing was performed by agar diffusion and the Phoenix automated method. Resistance genes were identified by PCR, and molecular typing of isolates was carried out by enterobacterial repetitive intergenic consensus (ERIC) sequences-polymerase chain reaction (PCR). ResultsAmong the 47 non-redundant MDR PA strains isolated, 59.57% were phenotypically ESBLs-positive, and 100% were phenotypically MBL-positive. The ESBL-positive isolates were subsequently screened for six groups of bla genes encoding ESBL-type enzymes, namely blaCTX-M2, blaPER, blaTEM, blaSHV, blaVEB, and blaGES. Out of the 28 ESBL-producing strains, 23 (82.14%) were blaCTX-M2 positive; 18 (38.29%) were blaPER positive, and 16 (34.04%) were blaTEM positive, while 5 (17.9%) were co-harboring blaCTX-M2, blaTEM, and blaPER genes. The blaSHV, blaVEB, and blaGES genes were not detected in any of the ESBL positive isolates. Since all isolates were MBL-positive, all 47 strains were screened for the blaNDM-1, blaIMP, blaVIM genes that produce MBLs; however, none of these genes were detected. Additional screening for the oprD gene demonstrated that 45 (95.74%) of the isolates were positive for this gene. Finally, ERIC PCR revealed 11 distinct PA clones among the blaCTX-M2 positive strains. ConclusionThis is the first study to report the presence of CTX-M2-producing PA in the North Africa region and the first to detect blaCTX-M2-positive and blaPER-positive PA clinical isolates in Algeria, therefore demonstrating the spread of such MDR strains to this part of the world. Identification of bacterial genotypic alterations that confer antibiotic resistance is critical in determining the most effective antimicrobial strategies to be employed. Therefore, our findings could potentially facilitate clinical decision making regarding the antibiotics of choice for the treatment of burn patients that suffer from PA infections in Algeria.

Rapid detection of extended spectrum β-lactamase producing Escherichia coli isolated from fresh pork meat and pig cecum samples using multiplex recombinase polymerase amplification and lateral flow strip analysis.

The emergence and dissemination of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli is a global health issue. Food-producing animals, including pigs, are significant reservoirs of antimicrobial resistance (AMR), which can be transmitted to humans. Thus, the rapid detection of ESBLs is required for efficient epidemiological control and treatment. In this study, multiplex recombinase polymerase amplification (RPA) combined with a single-stranded tag hybridization chromatographic printed-array strip (STH-PAS), as a lateral flow strip assay (LFA), was established for the rapid and simultaneous detection of multiple bla genes in a single reaction. Visible blue lines, indicating the presence of the blaCTX-M, blaSHV, and blaOXA genes, were observed within 10 min by the naked eye. The limit of detection of all three genes was 2.5 ng/25 μL, and no cross-reactivity with seven commensal aerobic bacteria was observed. A total of 93.9% (92/98) and 96% (48/50) of the E. coli isolates from pork meat and fecal samples, respectively, expressed an ESBL-producing phenotype. Nucleotide sequencing of the PCR amplicons showed that blaCTX-M was the most prevalent type (91.3-95.83%), of which the main form was blaCTX-M-55. The sensitivity and specificity of the RPA-LFA were 99.2% and 100%, respectively, and were in almost perfect agreement (κ = 0.949-1.000) with the results from PCR sequencing. Thus, the RPA-LFA is a promising tool for rapid and equipment-free ESBL detection and may facilitate clinical diagnosis in human and veterinary medicine, as well as AMR monitoring and surveillance.

Farmer’s Stance on Antibiotic Resistance to E. coli and Extended Spectrum - β -lactamase Producing (ESBL) E. coli Isolated from Poultry Droppings

Background: This study was conducted to explore the contribution of poultry farms to the contamination of the environment with ESBL-producing Escherichia coli and there with, potentially to the spread of these bacteria to humans and other animals. Hence, the present work is a poultry farm based study aimed to detect prevalence of ESBL producing E.coli among poultry of small scale farmers.Methods: ESBL-producing E. coli were detected at poultry farm (n=40). The E. coli was isolated from poultry droppings in irrespective of diseases. The required data were collected through well-structured interview schedule in farm premises. E.coli isolates were more susceptible to Gentamicin, Aztreonarm, Cefrtrazindime and Cefotoxime. Result: Detection of ESBL isolates was performed by Combined Disc Diffusion Methods. Out of 40 E.coli isolates 12 were phenotypically identified as ESBL producers. The prevalence of CTX-m gene is 50% and Bla (TEM) gene is 50%.

High fecal carriage of blaCTX-M, blaCMY-2, and plasmid-mediated quinolone resistance genes among healthy Korean people in a metagenomic analysis

To characterize the carriage of antibiotic resistance genes (ARGs) in the gut microbiome of healthy individuals. Fecal carriage of ARGs was investigated in 61 healthy individuals aged 30 to 59 years through whole metagenome sequencing of the gut microbiome and a targeted metagenomic approach. The number of ARGs in the gut microbiome was counted and normalized per million predicted genes (GPM). In the Korean population, the resistome ranged from 49.7 to 292.5 GPM (median 89.7). Based on the abundance of ARGs, the subjects were categorised into high (> 120 GPM), middle (60‒120 GPM), and low (< 60 GPM) ARG groups. Individuals in the high ARG group tended to visit hospitals more often (P = 0.065), particularly for upper respiratory tract infections (P = 0.066), and carried more blaCTX-M (P = 0.008). The targeted metagenome approach for bla and plasmid-mediated quinolone resistance (PMQR) genes revealed a high fecal carriage rate; 23% or 13.1% of the subjects carried blaCTX-M or blaCMY-2, respectively. Regarding PMQR genes, 59% of the subjects carried PMQR, and 83% of them harboured 2‒4 PMQR genes (qnrB 44.3%, qnrS 47.5% etc.). The presence of blaCTX-M correlated with ARG abundance in the gut resistome, whereas PMQR genes were irrelevant to other ARGs (P = 0.176). Fecal carriage of blaCTX-M and PMQR genes was broad and multiplexed among healthy individuals.

Seven-year surveillance of the prevalence of antimicrobial-resistant Escherichia coli isolates, with a focus on ST131 clones, among healthy people in Osaka, Japan

ObjectivesEscherichia coli (E. coli) is an indicator of antimicrobial resistance, and some strains of E. coli cause infectious diseases. E. coli sequence type 131 (ST131) – a global antimicrobial-resistant pandemic E. coli clone – is frequently detected in clinical specimens. Antimicrobial-resistant bacteria are monitored via national surveillance in clinical settings; however, monitoring information in non-clinical settings is limited. This study elucidated antimicrobial resistance trends of E. coli and dissemination of ST131 among healthy people in non-clinical settings. MethodsThis study collected 517 E. coli isolates from healthy people in Osaka, Japan, between 2013 and 2019. It analysed antimicrobial susceptibility of the isolates and detected the bla and mcr genes in ampicillin-resistant and colistin-resistant isolates, respectively, and the ST131 clone. ResultsAntimicrobial resistance rates of the bacteria isolated from healthy people in non-clinical settings were lower than for those in clinical settings. The resistance of the isolates to cefotaxime (4.4%) and ciprofloxacin (13.5%) gradually increased during the study period. In 23 cefotaxime-resistant isolates, the most frequent bla genes belonged to the blaCTX-M-9 group, followed by blaCTX-M-1 goup, blaTEM and blaCMY-2. One mcr-1-harbouring colistin-resistant isolate was detected in 2016. The incidence of the E. coli O25b-ST131 clone was approximately 5% until 2015 and 10% after 2016. ConclusionBoth ciprofloxacin resistance and O25b-ST131 clone frequency increased during the study period. Antimicrobial-resistant bacteria gradually spread in healthy people in non-clinical settings; one reason behind this spread was dissemination of global antimicrobial-resistant pandemic clones.

First evidence of blaNDM-1 and blaOXA-23 carbapenemase genes in human body lice infesting a second-hand T-shirt in a street market in Italy.

The spread of carbapenems resistance is a public health concern. The main group of carbapenemases encoding the β-lactamases activity (bla genes) is the Metallo-β-lactamases (MBLs). The presence of carbapenemase blaOXA-23-like, blaOXA-40-like, blaOXA-51-like, blaOXA-58-like, and blaNDM-1 genes was screened by real time PCR in 26 Pediculus humanus insects identified from second-hand clothes in a local market in Central Italy. Bacteria diversity was also characterized through shotgun metagenomic amplification for a deep sequencing of the host-associated bacterial microbiomes. The blaOXA-23 and blaNDM-1 carbapenemases genes were found and metagenomic analysis showed a great presence of Acinetobacter species. These results suggest a new potential transmission path for carbapenemase gene spread through bacteria ingested by insects infesting humans.