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122200.9415Molecular Characterization and the Antimicrobial Resistance Profile of Salmonella spp. Isolated from Ready-to-Eat Foods in Ouagadougou, Burkina Faso. The emergence of antimicrobial-resistantfood-borne bacteria is a great challenge to public health. This study was conducted to characterize and determine the resistance profile of Salmonella strains isolated from foods including sesames, ready-to-eat (RTE) salads, mango juices, and lettuce in Burkina Faso. One hundred and forty-eight biochemically identified Salmonella isolates were characterized by molecular amplification of Salmonella marker invA and spiC, misL, orfL, and pipD virulence genes. After that, all confirmed strains were examined for susceptibility to sixteen antimicrobials, and PCR amplifications were used to identify the following resistance genes: bla (TEM), temA, temB, StrA, aadA, sul1, sul2, tet(A), and tet(B). One hundred and eight isolates were genetically confirmed as Salmonella spp. Virulence genes were observed in 57.4%, 55.6%, 49.1%, and 38% isolates for pipD, SpiC, misL, and orfL, respectively. Isolates have shown moderate resistance to gentamycin (26.8%), ampicillin (22.2%), cefoxitin (19.4%), and nalidixic acid (18.5%). All isolates were sensitive to six antibiotics, including cefotaxime, ceftazidime, aztreonam, imipenem, meropenem, and ciprofloxacin. Among the 66 isolates resistant to at least one antibiotic, 11 (16.7%) were multidrug resistant. The Multiple Antimicrobial Resistance (MAR) index of Salmonella serovars ranged from 0.06 to 0.53. PCR detected 7 resistance genes (tet(A), tet(B), bla (TEM), temB, sul1, sul2, and aadA) in drug-resistant isolates. These findings raise serious concerns because ready-to-eat food in Burkina Faso could serve as a reservoir for spreading antimicrobial resistance genes worldwide.202236406904
123710.9396Characterization of Gene Families Encoding Beta-Lactamases of Gram-Negative Rods Isolated from Ready-to-Eat Vegetables in Mexico City. Beta-lactam resistant bacteria, which are commonly resident in tertiary hospitals, have emerged as a worldwide health problem because of ready-to-eat vegetable intake. We aimed to characterize the genes that provide resistance to beta-lactam antibiotics in Enterobacteriaceae, isolated from five commercial salad brands for human consumption in Mexico City. In total, twenty-five samples were collected, grown in blood agar plates, and the bacteria were biochemistry identified and antimicrobial susceptibility testing was done. The carried family genes were identified by endpoint PCR and the specific genes were confirmed with whole genome sequencing (WGS) by Next Generation Sequencing (NGS). Twelve positive cultures were identified and their microbiological distribution was as follows: 8.3% for Enterobacter aerogene (n = 1), 8.3% for Serratia fonticola (n = 1), 16.7% for Serratia marcesens (n = 2), 16.7% for Klebsiella pneumoniae (n = 2), and 50% (n = 6) for Enterobacter cloacae. The endpoint PCR results showed 11 colonies positive for blaBIL (91.7%), 11 for blaSHV (91.7%), 11 for blaCTX (97.7%), 12 for blaDHA (100%), four for blaVIM (33.3%), two for blaOXA (16.7%), two for blaIMP (16.7%), one for blaKPC (8.3%), and one for blaTEM (8.3%) gen; all samples were negative for blaROB, blaCMY, blaP, blaCFX and blaLAP gene. The sequencing analysis revealed a specific genotype for Enterobacter cloacae (blaSHV-12, blaCTX-M-15, blaDHA-1, blaKPC-2); Serratia marcescens (blaSHV-1, blaCTX-M-3, blaDHA-1, blaVIM-2); Klebsiella pneumoniae (blaSHV-12, blaCTX-M-15, blaDHA-1); Serratia fonticola (blaSHV-12, blaVIM-1, blaDHA-1); and, Enterobacter aerogene (blaSHV-1, blaCTX-M-1, blaDHA-1, blaVIM-2, blaOXA-9). Our results indicate that beta-lactam-resistant bacteria have acquired integrons with a different number of genes that provide pan-resistance to beta-lactam antibiotics, including penicillins, oxacillins, cefalosporins, monobactams, carbapenems, and imipenems.201830477153
122620.9391Multi-drug resistant gram-negative enteric bacteria isolated from flies at Chengdu Airport, China. We collected flies from Chengdu Shuangliu International Airport to examine for the presence of bacteria and to determine the sensitivity patterns of those bacteria. A total of 1,228 flies were collected from 6 sites around Chengdu Shuangliu International Airport from April to September 2011. The predominant species was Chrysomya megacephala (n=276, 22.5%). Antimicrobial-resistant gram-negative enteric bacteria (n=48) were isolated from flies using MacConkey agar supplemented with cephalothin (20 microg/ml). These were identified as Escherichia coli (n=37), Klebsiella pneumoniae (n=6), Pseudomonas aeruginosa (n=3) and Aeromonas hydrophila (n=2). All isolated bacteria were tested for resistance to 21 commonly used antimicrobials: amoxicillin (100%), ticarcillin (100%), cephalothin (100%), cefuroxime (100%), ceftazidime 1 (93.8%), piperacillin (93.8%), cefotaxime (89.6%), ticarcillin-clavulanate (81.3%), trimethoprim-sulfamethoxazole (62.5%), ciprofloxacin (54.2%), gentamicin (45.8%), cefepime (39.6%), tobramycin (39.6%), ceftazidime (22.9%), cefoxitin (16.7%), amikacin (16.7%), netilmicin (14.6%), amoxicillin-clavulanate (6.3%) and piperacillin-tazobactam (2.1%). No resistance to meropenem or imipenem was observed. Antibiotic resistance genes among the isolated bacteria were analyzed for by polymerase chain reaction. Thirty of the 48 bacteria with resistance (62.5%) possessed the blaTEM gene.201324450236
123330.9383Prevalence, Antibiogram, and Resistance Profile of Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Pig Farms in Luzon, Philippines. This cross-sectional study was conducted to determine the prevalence, antibiogram, and resistance profile of extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC) isolates from healthy pigs and pig farms in Luzon, Philippines. A total of 162 rectal samples from healthy finisher and breeder pigs and boot swab samples from pig houses were collected from 54 randomly selected pig farms. Bacteria were isolated and screened using MacConkey agar plate supplemented with 1 mg/L cefotaxime. Identification of bacteria and antimicrobial susceptibility test were carried out through Vitek(®) 2 and combined disk test. PCR amplifications were carried out in all isolates targeting bla(CTX-M) and its five major groupings, bla(TEM), and bla(SHV). The farm prevalence of ESBL-EC was 57.41% (95% confidence interval [CI] = 43.21-70.77). A total of 48 (29.63%) ESBL-EC isolates were isolated from samples that showed 14 different phenotypic multidrug resistance patterns. The prevalence of bla(CTX-M) gene was 91.67% (95% CI = 80.02-97.68). All major bla(CTX-M-groups) except bla(CTX-M-25group) were detected. The bla(CTX-M-1) was the most prevalent bla(CTX-M) gene, 75.0% (95% CI = 60.40-86.36). The prevalence of bla(TEM) and bla(SHV) genes was 91.67% (95% CI = 80.02-97.68) and 60.42% (95% CI = 45.27-74.23), respectively. Coexistence of different bla(CTX-M), bla(TEM), and bla(SHV) genes was observed in 44 isolates with 20 different genotypic patterns. High prevalence, diverse antibiogram profile, and genotypic resistance pattern of ESBL-EC isolates from healthy pigs and pig farms were observed in this study that could result in possible transmission to farm workers, susceptible bacteria, and the environment.202031532307
141340.9383Occurrence of Carbapenemases, Extended-Spectrum Beta-Lactamases and AmpCs among Beta-Lactamase-Producing Gram-Negative Bacteria from Clinical Sources in Accra, Ghana. Beta-lactamase (β-lactamase)-producing Gram-negative bacteria (GNB) are of public health concern due to their resistance to routine antimicrobials. We investigated the antimicrobial resistance and occurrence of carbapenemases, extended-spectrum β-lactamases (ESBLs) and AmpCs among GNB from clinical sources. GNB were identified using matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDITOF-MS). Antimicrobial susceptibility testing was performed via Kirby-Bauer disk diffusion and a microscan autoSCAN system. β-lactamase genes were determined via multiplex polymerase chain reactions. Of the 181 archived GNB analyzed, Escherichia coli and Klebsiella pneumoniae constituted 46% (n = 83) and 17% (n = 30), respectively. Resistance to ampicillin (51%), third-generation cephalosporins (21%), and ertapenem (21%) was observed among the isolates, with 44% being multi-drug resistant (MDR). β-lactamase genes such as AmpCs ((bla(FOX-M) (64%) and bla(DHA-M) and bla(EDC-M) (27%)), ESBLs ((bla(CTX-M) (81%), other β-lactamase genes bla(TEM) (73%) and bla(SHV) (27%)) and carbapenemase ((bla(OXA-)(48) (60%) and bla(NDM) and bla(KPC) (40%)) were also detected. One K. pneumoniae co-harbored AmpC (bla(FOX-M) and bla(EBC-M)) and carbapenemase (bla(KPC) and bla(OXA-)(48)) genes. bla(OXA-)(48) gene was detected in one carbapenem-resistant Acinetobacter baumannii. Overall, isolates were resistant to a wide range of antimicrobials including last-line treatment options. This underpins the need for continuous surveillance for effective management of infections caused by these pathogens in our settings.202337370334
141750.9382Prevalence and Phenotypic and Molecular Characterization of Carbapenemase-Producing Gram-Negative Bacteria in Gabon. Data collection and monitoring of carbapenemase-producing (CP) Gram-negative bacteria (GNB) are often limited. This study determined CP-GNB prevalence in Gabon and the genetic origins of the resistance genes. From January 2016 to March 2018, 869 clinically significant GNB isolates from inpatients and outpatients, and 19 fecal samples (inpatients) were analyzed in the main hospitals of Gabon. Fecal samples were screened using ChromID® CARBA SMART selective chromogenic medium biplates. Species were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Antibiotic susceptibility was tested using the disk diffusion method on Müller-Hinton agar, and resistance genes were assessed by multiplex polymerase chain reaction and sequencing. Overall, 1.61% of clinical isolates (14 of 869) and 5.26% of fecal samples (1 of 19) were CP-GNB. The CP-GNB rate was higher among inpatients (2.98%) than outpatients (0.33%), in intensive care units (28.57%, 4 of 14), and in urine samples (35.71%, 5 of 14). The most common CP-GNB were Klebsiella pneumoniae (53.33%) and Acinetobacter baumannii (26.67%). blaOXA-48 was the predominant carbapenemase-encoding gene (40%), followed by blaNDM-5 (33.33%). The A. baumannii multilocus sequence types ST2 and ST78, Enterobacter cloacae ST78, Escherichia coli ST2, and K. pneumonia ST48 and ST147 were found. These data indicate that CP bacteria are present in clinical and carriage samples. Preventive measures are needed to avoid the spread of resistance genes.202336535247
141260.9382A highly multiplexed melt-curve assay for detecting the most prevalent carbapenemase, ESBL, and AmpC genes. Resistance to third-generation cephalosporins and carbapenems in Gram-negative bacteria is chiefly mediated by beta-lactamases including extended-spectrum beta-lactamase (ESBL), AmpC, and carbapenemase enzymes. Routine phenotypic detection methods do not provide timely results, and there is a lack of comprehensive molecular panels covering all important markers. An ESBL/carbapenemase high-resolution melt analysis (HRM) assay (SHV, TEM, CTX-M ESBL families, and NDM, IMP, KPC, VIM and OXA-48-like carbapenemases) and an AmpC HRM assay (16S rDNA control, FOX, MOX, ACC, EBC, CIT, and DHA) were designed and evaluated on 111 Gram-negative isolates with mixed resistance patterns. The sensitivity for carbapenemase, ESBL, and AmpC genes was 96.7% (95% confidence interval [CI]: 82.8-99.9%), 93.6% (95% CI: 85.7-97.9%), and 93.8% (95% CI: 82.8-98.7%), respectively, with a specificity of 100% (95% CI: 95.6-100%), 93.9% (95% CI: 79.8-99.3%), and 93.7% (95% CI: 84.5-98.2%). The HRM assays enable the simultaneous detection of the 14 most important ESBL, carbapenemase, and AmpC genes and could be used as a molecular surveillance tool or to hasten detection of antimicrobial resistance for treatment management.202032521424
141170.9381Detection and characterization of carbapenem resistant Gram-negative bacilli isolates recovered from hospitalized patients at Soba University Hospital, Sudan. BACKGROUND: Antimicrobial resistance (AMR) poses a complex threat to global health security and universal health coverage. Recently, nosocomial infections with carbapenemase-producing Gram-negative bacilli (GNB) is increasing worldwide. We report the molecular characterization and detection of genes associated with carbapenemase producing Gram negative bacteria isolated from hospitalized patients at Soba University Hospital (SUH) in Khartoum State, Sudan. RESULTS: Between October 2016 and February 2017, a total of 206 GNB clinical specimens were collected from hospitalized patients in SUH. Of 206 carbapenem resistance isolates, 171 (83 %) were confirmed as phenotypically resistant and 121 (58.7 %) isolates harboured one or more carbapenemase genes. New Delhi metallo-β-lactamase (NDM) types were the most predominant genes, blaNDM 107(52 %), followed by blaIMP 7 (3.4 %), blaOXA-48 5(2.4 %) and blaVIM 2 (0.9 %). Co-resistance genes with NDM producing GNB were detected in 87 (81.3 %) of all blaNDM producing isolates. NDM-1 was the most frequent subtype observed in 75 (70 %) blaNDM producing isolates. The highest percentage of resistance was recorded in ampicillin (98 %), cephalexin (93.5 %) amoxicillin clavulanic acid (90 %), cefotaxime (89.7 %), ceftriaxone (88.4 %), ceftazidime (84.2 %), sulfamethoxazole-trimethoprim (78.4 %) and nitrofurantoin (75.2 %), aztreonam (66 %) and temocillin (64 %). A close correlation between phenotypic and carbapenemase genes detection in all GNB was observed. CONCLUSIONS: The frequency of carbapenemase producing bacilli was found to be high in SUH. NDM was found to be the most prevalent carbapenemase gene among clinical isolates. Close surveillance across all hospitals in Sudan is required. The relative distribution of carbapenemase genes among GNB in nosocomial infections in Africa needs to be defined.202133947325
145580.9379Resistance to bacterial infection, complication occurring after cardiac surgery. To analyze the occurrence of resistant bacterial infection in patients undergoing cardiac surgery hospitalized in the surgical specialty hospital, in Erbil city, Iraq. A prospective study was done on a total of 138 patients operated and hospitalized in an intensive care unit and surgical wards. Bacterial isolates identification was done according to cultural characteristics, microscopic examination, some biochemical tests, analytic Profile Index 20E& API Staph, confirmed with VITEK® 2 compact system (BioMérieux). Antimicrobial susceptibility for disc diffusion tested to 17 antimicrobial agents. Resistance isolates were confirmed phenotypically for carbapenemase by Rapidec Carba NP Test (bioMe´rieux SA, Marcy-l'E´toile, France) for ESBLs producers by ESBL screening test VITEK 2 system. Molecularly blaIMP blaTEM, blaKPC, AmpC and blaCTX-M were detected by PCR. In 134 patients, 28.3% of patients got infected post-operatively. The most frequent source of isolation was from ICU patients (75%). Isolated bacteria included gram-positive 29 (54.7%) and gram-negative bacteria 24 (45.3%). Most frequently:  Staphylococcus aureus (24.4%), each of pseudomonas aeroginosa, Klebsiella pneumonia (15.1%), Streptococcus spp. (11.3%), Escherichia coli (9.4%). Whereas included Coagulase Negative Staphylococci species (CoNS) (13.2%) and Enterococci species (5.7) Statistical analysis showed significantly higher sensitive isolates as compared with resistance isolates. Resistance to Carbapenems calss was 18.9% and Cephalosporins class 41.5% of isolates. The antimicrobial resistance pattern indicated that MDR bacterial isolates (81.1%) were widespread. Of the 34 phenotypically ESBL positive isolates, the ESBL genes (AmpC, blaCTX-M, and blaTEM) were amplified in 7(20.6), 6(17.6) and 6(17.6) isolates respectively. Out of 8 K. pneumonia (37.5%) harboring both blaAmpC and bla-CTX-M genes, while 6(75%) carries blaTEM. The blaCTX-M gene was found in only 1 (12.5%) out of 8 isolates of P. aeruginosa. While blaAmpC genotyping revealed that 1(7.7%) out of 13 Staph. aureus isolates were harboring it. Finally, 3(60%) out of 5 E. coli isolates harboring both AmpC and bla-CTX-M genes. Cardiac surgery patients wound show increasingly emerging strains of ESBL-producing gram-negative bacteria K. pneumonia, P. aeruginosa and E. coli especially patients prolonged in the intensive care unit.202034174972
123190.9376Prevalence and Molecular Characterization of Plasmid-mediated Extended-Spectrum β-Lactamase Genes (balaTEM, blaCTX and blASHV) Among Urinary Escherichia coli Clinical Isolates in Mashhad, Iran. OBJECTIVES: Extended-spectrum beta-lactamase (ESBL) producing bacteria have an important role in nosocomial infections. Due to the limited availability of information about the molecular epidemiology of ESBL producing bacteria in Mashhad, we decided to investigate about TEM, CTX and SHV ESBLs among urinary Escherichia coli isolates in Mashhad, a city in northeast Iran. MATERIALS AND METHODS: One hundred and eleven clinical isolates of E. coli were diagnosed from hospitalized patients in 2009. After performing antibiogram and phenotypic confirmation test, polymerase chain reaction (PCR) was performed by blaTEM, blaSHV and blaCTX primers and restriction digestion was carried out using PstI and TaqI (Fermentas-Lithuania) for confirmation. RESULTS: ESBL producers of E. coli isolates were 33.3%. Among 37 ESBL-producing isolates, 35 (94.6%), 21 (56.8%) and 5 (13.5%) were shown to have blaCTX, blaTEM and blaSHV, genes respectively. Co-resistance to non-beta lactam antibiotics was observed more with ESBL producers (P < 0.05). CONCLUSION: The results showed that the studied ESBL genes are found with high prevalence and among them blaCTX is more widespread in urine E. coli isolates in Mashhad.201223493415
2094100.9376Evaluation of the antibacterial activity of Weissella confusa K3 cell-free supernatant against extended-spectrum βeta lactamase (ESBL) producing uropathogenic Escherichia coli U60. Different strategies have been approved for controlling extended-spectrum βeta lactamase (ESBL) producing uropathogenic bacteria. The antibacterial activity of Lactic acid bacteria (LAB) is an effective strategy due to its probiotic characteristics and beneficial effects on human health. The antibiotic susceptibility test, disk diffusion method, and double disc synergy test indicated that five enteric uropathogenic isolates were ESBL producers during the present study. They recorded diameters of inhibition zones as ≤ 18, ≤ 8, ≤ 19, and ≤ 8 mm against cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), and ceftriaxone (CRO). Genotypically, bla(TEM) genes are the most common, with (100 %) occurrence in all the five enteric tested uropathogens, followed by bla(SHV) and bla(CTX) genes (60 %). In addition, out of 10 LAB isolates from dairy products, the CFS of isolate no. K3 had high antibacterial activity against the tested ESBLs, especially no. U60, with a MIC of 600 µl. Additionally, the MIC and sub-MIC of K3 CFS inhibited the production of antibiotic-resistant bla (TEM) genes of U60. Analyzing the 16S rRNA sequence confirmed that the most potent ESBL-producing bacteria (U60) and LAB (K3) isolates were identified as Escherichia coli U60.1 and Weissella confuse K3 with accession numbers MW173246 and MW173299.1, respectively, in GenBank.202336873575
1426110.9376Phenotypic and genotypic detection of carbapenemase production among gram negative bacteria isolated from hospital acquired infections. OBJECTIVES: To identify the carbapenemase producing Gram-negative bacteria (GNB) by phenotypic methods and to confirm the presence of resistant genes using real-time polymerase chain reaction (PCR). METHODS: This was a prospective study carried out at the Department of Microbiology, Sri Venkata Sai Medical College and Hospital, Mahabubnagar, India, from March 2018-2021. All samples were screened for carbapenem resistance by disc diffusion method and the VITEK(®)2 compact system (bioMérieux, France). Detection of carbapenemase was carried out using RAPIDEC(®)CARBA NP test (Biomeriux Private Limited, South Delhi, India), screening for metallo-β-lactamases (MBL) was carried out by double disk synergy test (DDST), and genotypic characterization by real-time PCR. RESULTS: Among the 1093 Gram-negative bacilli identified, 220 (17.0%) were resistant to carbapenems by both tested methods. Carbapenemase detection using the RAPIDEC(®)CARBA NP test indicated that 207 (94.0%) were carbapenemase producers, of which 189 (91.2%) were MBL producers. The most common carbapenemase genes identified were New Delhi metallo-β-lactamase (NDM; 47.3%), followed by the co-existence of genes in combination of NDM, with Verona integron-mediated metallo-β-lactamase (VIM; 39.6%), VIM and oxacillin hydrolyzing enzymes-48 (OXA-48; 4.3%), and OXA-48 (1.4%).No gene of active on imipenem, Klebsiella pneumonia carbapenemase, VIM, or OXA-48 alone was detected. CONCLUSION: This study suggests routine carbapenem resistance testing among multi-drug resistant-GNBs, as most of these infections occur in hospitals. In addition, there is a possibility that these highly antibiotic-resistant genes could spread to other bacteria resulting in further dissemination.202235256490
1415120.9375Antibiogram and Molecular Characterization of AmpC and ESBL-Producing Gram-Negative Bacteria from Poultry and Abattoir Samples. BACKGROUND AND OBJECTIVE: The global antibiotic resistance threat posed by ESBL and AmpC-producing Gram-Negative Bacteria (GNB) is a public health menace that rolls back the gains of 'One Health'. This study investigated the antibiogram and prevalence of AmpC and ESBL genes in Escherichia coli, Klebsiella spp. and Pseudomonas spp. from poultry and abattoir milieus in Enugu and Ebonyi States, Nigeria. MATERIALS AND METHODS: Isolation, identification and characterization of GNB from samples (150 abattoirs and 300 poultry) were done using standard microbiological techniques. Antimicrobial Susceptibility Testing (AST), as well as phenotypic screening for ESBL and AmpC enzymes, was performed using the Kirby-Bauer disc diffusion technique. PCR technique was used to screen isolated GNB for AmpC and ESBL genes. RESULTS: Exactly 42 E. coli and 8 Klebsiella spp. isolate from poultry samples and another 5 P. aeruginosa isolates from abattoir samples were phenotypically confirmed to be ESBL-producers. AmpC enzymes were phenotypically detected in 8 E. coli and 13 P. aeruginosa isolates from poultry samples. All ESBL and AmpC-positive bacteria exhibited high resistance frequencies to tested antibiotics, especially to the carbapenems and cephalosporins. ESBL genes (CTX-M, SHV-1, TEM) and AmpC genes (ACC-M, MOX-M, DHA-M) were harbored by the isolated GNB in this study. Overall, the DHA-M and CTX-M genes, mediating AmpC and ESBL production respectively were the most prevalent genes harbored by the tested GNB. CONCLUSION: This study reported that AmpC and ESBL genes are harbored by Gram-negative bacteria (E. coli, Klebsiella species and P. aeruginosa) that emanated from poultry and abattoir milieus.202133683048
1427130.9375Prevalence and Characterization of Carbapenem-Resistant Enterobacteriaceae Isolated from Mulago National Referral Hospital, Uganda. INTRODUCTION: Carbapenemases have increasingly been reported in enterobacteriaceae worldwide. Most carbapenemases are plasmid encoded hence resistance can easily spread. Carbapenem-resistant enterobacteriaceae are reported to cause mortality in up to 50% of patients who acquire bloodstream infections. We set out to determine the burden of carbapenem resistance as well as establish genes encoding for carbapenemases in enterobacteriaceae clinical isolates obtained from Mulago National Referral Hospital, Uganda. METHODS: This was a cross-sectional study with a total of 196 clinical isolates previously collected from pus swabs, urine, blood, sputum, tracheal aspirates, cervical swabs, endomentrial aspirates, rectal swabs, Vaginal swabs, ear swabs, products of conception, wound biopsy and amniotic fluid. All isolates were subjected to phenotypic carbapenemase screening using Boronic acid-based inhibition, Modified Hodge and EDTA double combined disk test. In addition, all the isolates were subjected to PCR assay to confirm presence of carbapenemase encoding genes. RESULTS: The study found carbapenemase prevalence of 22.4% (44/196) in the isolates using phenotypic tests, with the genotypic prevalence slightly higher at 28.6% (56/196). Over all, the most prevalent gene was blaVIM (21,10.7%), followed by blaOXA-48 (19, 9.7%), blaIMP (12, 6.1%), blaKPC (10, 5.1%) and blaNDM-1 (5, 2.6%). Among 56 isolates positive for 67 carbapenemase encoding genes, Klebsiella pneumonia was the species with the highest number (52.2%). Most 32/67(47.7%) of these resistance genes were in bacteria isolated from pus swabs. CONCLUSION: There is a high prevalence of carbapenemases and carbapenem-resistance encoding genes among third generation cephalosporins resistant Enterobacteriaceae in Uganda, indicating a danger of limited treatment options in this setting in the near future.201526284519
1244140.9375Identification of antibiotic resistance genes in Escherichia coli from subclinical mastitis milk in dairy cows and goats, East Java Province. Antibiotics are still used to treat mastitis in dairy cows in Indonesia. This study aimed to analyse antibiotic resistance genes in Escherichia coli (E. coli) from subclinical mastitis milk in East Java Province, Indonesia. The samples consisted of subclinical mastitis milk from cows and goats. A total of 592-quarter cow's milk and 71 goat's milk samples from both halves of the udder were collected from 67 farms in Lumajang, Banyuwangi, Malang, Sidoarjo, Jember, Pasuruan, Probolinggo, and Mojokerto. Subclinical mastitis samples were screened using the California mastitis test (CMT). E. coli was identified by phenotypic and genotypic methods. E. coli was confirmed with a primer specific to the polymerase chain reaction (PCR) technique. Gene resistance of E. coli was tested using the multiplex-PCR (mPCR) technique with primers encoding the genes temoneira enzyme (TEM), oxacillinase (OXA), sulfhydryl variable (SHV), and cefotaximase-munich IV (CTX-M IV). These genes were chosen because mastitis treatment generally uses oxacilline and β-lactam antibiotics. All data obtained were analysed descriptively. The results show that six isolates of E. coli (46.15%) carried a single resistance gene (TEM or SHV) and two isolates (33.33%) were confirmed as multiple drug-resistant organisms (MDROs) (TEM and SHV). The resistance genes were found in samples originating from Blitar, Banyuwangi, Lumajang, and Pasuruan Regencies. This research implies that antibiotic-resistance genes found in E. coli on certain farms are dangerous and may allow gene transmission to other bacteria that make treatment for mastitis or other bacterial infections ineffective.202438550619
6054150.9372Lactobacillus Strains for Vegetable Juice Fermentation-Quality and Health Aspects. Vegetable juices are new carrier variants for beneficial bacteria, representing an alternative to dairy-fermented products, especially for vegan, strict vegetarian, or allergic consumers. The aim of this study was to characterize several Romanian native lactic acid bacteria (LAB) strains to select valuable nutritional and probiotic strains for vegetable juice fermentation. Nineteen LAB strains were analyzed for antibiotic susceptibility (disc-diffusion method), the presence of antibiotic resistance genes, the presence of functional genes. and the production of organic acids by HPLC. Antibiotic resistant strains were observed only with ampicillin (Amp10) and kanamycin (K30), 79% and 32%, respectively, with results partially confirmed by molecular analysis. Multiplex PCR revealed the presence of LBA1272, dltD, folP, agl, α-amy, malL, and ribA genes, related to stress resistance, starch metabolism, and production of vitamins, except for folK. HPLC analyses were performed on beet roots (SF), tomato (TM), and a mixture of carrots, celery, and beet (MTS) juices. High values of lactic acid were recorded in all cases of LAB fermentation (5034-14,176 µg/mL). The maximum values recorded for acetic acid did not exceed 2.5 mg/mL having a positive influence on the product's taste.202236359394
1418160.9372Nosocomial infections and antimicrobial susceptibility patterns among patients admitted to intensive care unit of Imam Khomeini hospital in Ilam, Iran. INTRODUCTION: Nosocomial infections (NIs) are a major challenge worldwide. Identification of antibiotic resistance pattern extended spectrum beta-lactamases (ESBLs) and carbapenem-resistant Enterobacteriaceae (CRE) were the objectives of this study. METHODS: In this cross-sectional study, the antimicrobial susceptibility pattern of bacterial isolates collected from patients with NIs in ICU was determined. Overall, 42 Escherichia coli and Klebsiella pneumoniae isolates from different infection sites were used to determine phenotypic tests of ESBLs, Metallo-β-lactamases (MBLs) and CRE. Detection of ESBLs, MBLs and CRE genes were performed by the polymerase chain reaction (PCR) method. RESULTS: From 71 patients with NIs, 103 different bacterial strains were isolated. The most frequently isolated bacteria were E. coli (n = 29; 28.16%), Acinetobacter baumannii (n = 15; 14.56%), and K. pneumoniae (n = 13; 12.26%). Also, the rate of multidrug-resistant (MDR) isolates was 58.25% (60/103). Based on phenotypic confirmation tests, 32 (76.19%) isolates of E. coli and K. pneumoniae produced ESBLs, and 6 (14.28%) isolates were identified as CRE producers. PCR showed the high prevalence of the bla(CTX-M) (n = 29; 90.62%) in ESBL genes. In addition, bla(NDM) was detected in 4 (66.66%), bla(OXA-23) in 3 (50%), and bla(OXA-48) gene in 1 (16.66%) isolates. The bla(VIM), bla(KPC), and bla(IMP) genes were not detected in any of the isolates. CONCLUSION: The Gram-negative bacteria E. coli, A. baumannii, and K. pneumoniae with high resistance levels were the most common bacteria causing NIs in the ICU. This study for the first time identified bla(OXA-11), bla(OXA-23), and bla(NDM-1) genes in E. coli and K. pneumoniae in Ilam city of Iran.202337155016
1221170.9372Invasive whistling frogs (Eleutherodactylus johnstonei) act as a reservoir for antimicrobial-resistant Enterobacteriaceae in Latin America's most populous city. Invasive species represent a significant threat to ecological balance and the maintenance of native populations. Besides, these have been associated with the emergence of pathogens of public health importance, including multidrug-resistant bacteria. This study aimed to screen and describe the antimicrobial resistance profile of clinically important Enterobacteriaceae species isolated from whistling frogs (Eleutherodactylus johnstonei), an invasive anuran species in São Paulo, Brazil. Clinically relevant Enterobacteriaceae strains (n = 35) were isolated from oral and skin swabs of 19 whistling frogs and tested for antimicrobial susceptibility and antimicrobial resistance encoding genes. Resistance to amoxicillin + clavulanate and cefoxitin were the most frequent (16.67%; 4/24), followed by cefotaxime (5.71%; 2/35), ceftriaxone (2.86%; 1/35), and tetracycline (2.86%; 1/35). Among the antimicrobial resistance genes screened, bla(CTX-M group 8), bla(TEM), and bla(CMY) were identified. The whole genome of the bla(CTX-M group 8)-positive E. coli strain was assessed and confirmed bla(CTX-M-8) presence and phylogenetic analysis. Given the synanthropic behavior of whistling frogs, these amphibians may act as carriers of antimicrobial-resistant bacteria.202540884707
1436180.9370Characterisation of carbapenem-resistant Gram-negative organisms from clinical specimens in Yola, Nigeria. OBJECTIVES: This study aimed to identify carbapenem-resistant Gram-negative bacteria from clinical specimens of patients in Yola, Nigeria. METHODS: Routine clinical specimens were screened for the presence of carbapenem-resistant Gram-negative bacteria using chromogenic agar plates. Susceptibility of all presumptive isolates to carbapenems was tested by MIC and disk diffusion methods. Real-time PCR was used to test for the presence of carbapenemase genes. RESULTS: Screening of 1741 clinical specimens yielded 119 (6.8%) presumptive carbapenem-resistant Gram-negative bacteria. Antimicrobial susceptibility testing confirmed carbapenem resistance in 105 of these isolates. New Delhi metallo-β-lactamase (bla(NDM)) gene was detected in 26 isolates and Verona integron-encoded metallo-β-lactamase (bla(VIM)) gene was detected in four. The mechanism of resistance could not be identified in approximately two thirds of the carbapenem-resistant isolates. CONCLUSION: While bla(NDM) and bla(VIM) accounted for 28.6% of the resistance seen, further molecular-based studies are needed to characterise the other mechanisms of carbapenem resistance in these isolates.202031472281
1464190.9370Detection of TEM and CTX-M genes from ciprofloxacin resistant Proteus mirabilis and Escherichia coli isolated on urinary tract infections (UTIs). The multidrug resistant Gram negative bacteria (MDRGNB) is an emerging burden and now represents a daily challenge for the management of antimicrobial therapy in healthcare settings. The present study was aimed to detect the prevalence of TEM and CTX-M type genes from GNB on urinary tract infection (UTIs). The ciprofloxacin resistant uropathogens were detected by HEXA UTI 5 disc diffusion method. The phenotypic detection of uropathogens producing extended spectrum beta lactamases (ESBLs) was confirmed by double disc combination test (DDCT) and phenotype confirmation test (PCT). The prevalence of TEM and CTX-M genes of uropathogens was identified by multiplex PCR analysis. The in vitro antimicrobial susceptibility of E. coli producing ESBL (26), 21 isolates of P. mirabilis, 17 P. aeruginosa, 14 K. pneumoniae and 6 Enterobacter sp. were detected. Based on the extension of the cephalosporin zone edge towards augmentin disc in the DDST method proved 84% of the isolates were ESBL positive. Similar results were obtained in phenotypic confirmatory test (PCT) by the increases of ≥5 mm zone of inhibition in the combination disc when compared with ceftazidime disc alone. The prevalence of TEM and CTX-M genes were determined from multidrug resistance uropathogens (MDU) respectively as 83%, 75%, 71%, 63%, 60%, 55%, 54%, 50%. The most prevalent (TEM + CTX-M) genes were also detected in ciprofloxacin resistant strains P. mirabilis BDUMS1 (KY617768) and E. coli BDUMS3 (KY617770). Due to the increase of ESBL genes in uropathogens, sustained supervision for using favorable antibiotics and decreasing the infection is essential.201829778819