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125300.9435Phenotypic and Genotypic Assessment of Antibiotic Resistance and Genotyping of vacA, cagA, iceA, oipA, cagE, and babA2 Alleles of Helicobacter pylori Bacteria Isolated from Raw Meat. BACKGROUND: Foodstuffs with animal origins, particularly meat, are likely reservoirs of Helicobacter pylori. PURPOSE: An existing survey was accompanied to assess phenotypic and genotypic profiles of antibiotic resistance and genotyping of vacA, cagA, cagE, iceA, oipA, and babA2 alleles amongst the H. pylori bacteria recovered from raw meat. METHODS: Six-hundred raw meat samples were collected and cultured. H. pylori isolates were tested using disk diffusion and PCR identification of antibiotic resistance genes and genotyping. RESULTS: Fifty-two out of 600 (8.66%) raw meat samples were contaminated with H. pylori. Raw ovine meat (13.07%) had the uppermost contamination. H. pylori bacteria displayed the uppermost incidence of resistance toward tetracycline (82.69%), erythromycin (80.76%), trimethoprim (65.38%), levofloxacin (63.46%), and amoxicillin (63.46%). All H. pylori bacteria had at least resistance toward one antibiotic, even though incidence of resistance toward more than eight antibiotics was 28.84%. Total distribution of rdxA, pbp1A, gyrA, and cla antibiotic resistance genes were 59.61%, 51.92%, 69.23%, and 65.38%, respectively. VacA s1a (84.61%), s2 (76.92%), m1a (50%), m2 (39.13%), iceA1 (38.46%), and cagA (55.76%) were the most generally perceived alleles. S1am1a (63.46%), s2m1a (53.84%), s1am2 (51.92%), and s2m2 (42.30%) were the most generally perceived genotyping patterns. Frequency of cagA-, oipA-, and babA2- genotypes were 44.23%, 73.07%, and 80.76%, respectively. A total of 196 combined genotyping patterns were also perceived. CONCLUSION: The role of raw meat, particularly ovine meat, in transmission of virulent and resistant H. pylori bacteria was determined. VacA and cagA genotypes had the higher incidence. CagE-, babA2-, and oipA- H. pylori bacteria had the higher distribution. Supplementary surveys are compulsory to originate momentous relations between distribution of genotypes, antibiotic resistance, and antibiotic resistance genes.202032099418
94810.9426Multidrug-Resistant Bacteria in Aquaculture Systems in Accra, Ghana. BACKGROUND: Antibiotic resistance (ABR) poses a critical global health challenge, necessitating its surveillance across both human and animal health sectors. This study evaluated ABR in bacteria harboured in reared inland fishes sold in Accra and the pond water from which they originated. METHOD: The study was cross-sectional, involving fishes and water sampled from 80 ponds. The gastrointestinal organs of the fishes were homogenised and cultured for bacteria, as were the water samples. The bacteria were identified using matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF-MS). Antimicrobial susceptibility test was done using the Kirby-Bauer method. Multidrug-resistant (MDR) bacteria were selected for further testing. The double disc diffusion method was used to detect extended-spectrum beta-lactamase (ESBL) production in isolates that were resistant to third-generation cephalosporins. Whole genome sequencing was performed on the ESBL-positive isolates using the Illumina Miseq platform. RESULTS: In total, 39 different bacterial species, with their individual numbers totalling 391, were isolated. The bacteria were predominantly Escherichia coli (17%), Aeromonas veronii (11%), Citrobacter freundii (8%), Bacillus cereus (5%), and Klebsiella pneumoniae (5%). The overall ABR rates were cefotaxime (32%), gentamicin (1%), ciprofloxacin (4%), chloramphenicol (19%), tetracycline (37%), meropenem (0%), and ertapenem (0%). Overall MDR and ESBL bacteria prevalence were 13.6% and 1.3%, respectively. The sequence types of the ESBL isolates were ST4684 (80%, n = 4) and ST2005 (20%, n = 1), and the serotypes were H34:09 (80%, n = 4) and H7 (20%, n = 1); the ABR genes were blaCTX-M-15, fosA7, and qnrS1. CONCLUSION: The fishes and the pond water were contaminated with a diverse range of bacteria, mainly Escherichia coli and Aeromonas veronii. The ABR, MDR, and ESBL rates were low to moderate. Moreover, the main sequence type and serotype of the ESBL isolates were ST4684 and H34:09, respectively, and the ABR genes were blaCTX-M-15, fosA7, and qnrS1.202439600552
526320.9419Seasonal Variations in Water-Quality, Antibiotic Residues, Resistant Bacteria and Antibiotic Resistance Genes of Escherichia coli Isolates from Water and Sediments of the Kshipra River in Central India. OBJECTIVES: To characterize the seasonal variation, over one year, in water-quality, antibiotic residue levels, antibiotic resistance genes and antibiotic resistance in Escherichia coli isolates from water and sediment of the Kshipra River in Central India. METHODS: Water and sediment samples were collected from seven selected points from the Kshipra River in the Indian city of Ujjain in the summer, rainy season, autumn and winter seasons in 2014. Water quality parameters (physical, chemical and microbiological) were analyzed using standard methods. High-performance liquid chromatography⁻tandem mass spectrometry was used to determine the concentrations of antibiotic residues. In river water and sediment samples, antibiotic resistance and multidrug resistance patterns of isolated E. coli to 17 antibiotics were tested and genes coding for resistance and phylogenetic groups were detected using multiplex polymerase chain reaction. One-way analysis of variance (ANOVA) and Fisher tests were applied to determine seasonal variation. RESULTS: In river water, seasonal variation was significantly associated with various water quality parameters, presence of sulfamethoxazole residues, bacteria resistant to ampicillin, cefepime, meropenem, amikacin, gentamicin, tigecycline, multidrug resistance and CTX-M-1 gene. The majority of the Extended Spectrum Beta-Lactamase (ESBL)-producing E. coli isolates from river water and sediment in all different seasons belonged to phylogenetic group A or B1. CONCLUSIONS: Antibiotic pollution, resistance and resistance genes in the Kshipra River showed significant seasonal variation. Guidelines and regulatory standards are needed to control environmental dissemination of these “pollutants” in this holy river.201829914198
141530.9417Antibiogram 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
210040.9414Prevalence of Bacteria and Antimicrobial Resistance Genes in Hospital Water and Surfaces. Purpose Antimicrobial resistance (AMR) has become a worldwide environmental and public health problem, causing more than 250,000 deaths per year. Unregulated usage, unsafe hospital practices, and misuse in veterinary contribute to the development of multidrug resistance in various bacteria. Hospital water was hypothesized to be a hotspot for AMR transmission because of (1) increased exposure to antibiotic load, (2) poor drainage and sanitation system, (3) interaction between environmental and clinical microbes. The purpose of the research was to assess the biodiversity and AMR in hospital tap waters. Methodology In this study, the microflora of the hospital tap water and hospital surfaces was observed by obtaining water samples from the intensive care unit (ICU), surgical wards, and washrooms. These were processed through membrane filtration and spread on seven different media (Aeromonas Medium, Azide Dextrose Agar, MacConkey Agar, Mannitol Salt Agar, Pseudomonas Cetrimide Agar, Salmonella Shigella Agar, and Thiosulfate Citrate Bile Salts Sucrose Agar). Surface samples were collected from the faucet, basin, and drain and directly spread on the media plates. Isolates were identified using standard bacteriological and biochemical tests. Kirby-Bauer disk diffusion method was performed using 21 antibiotic disks from 10 different antibiotic classes. They included ampicillin (AMP), amoxicillin (AML), piperacillin-tazobactam (TZP), cefipime (FEP), cefoxitin (FOX), ceftazidime (CAZ), ceftriaxone (CRO), imipenem (IMP), meropenem (MEM), ciprofloxacin (CIP), moxifloxacin (MXF), levofloxacin (LEV), amikacin (AK), gentamicin (CN), tigecycline (TGC), aztreonam (ATM), erythromycin (E), clindamycin (DA), rifampicin (RD), colistin (CT), and chloramphenicol (C). The results were interpreted according to EUCAST guidelines for the antibiogram of the isolates; 38 isolates were selected out of 162 based on different parameters for genotyping and detection of six beta-lactamase genes (blaSHV, blaTEM, blaCTX-M, blaOXA, blaKPC, blaNDM). Results Among these 162 isolates, 82 were obtained from water sources and 80 were collected from surfaces (faucet, basin, drain). The isolates included a variety of bacteria including Aeromonas spp. (20%), Klebsiella spp. (13%), Staphylococcus aureus (13%), Pseudomonas spp.(10%), Escherichia coli (9%), Vibrio spp. (8%), Enterococcus spp. (6%), Shigella spp. (6%), Salmonella spp. (4%), Acinetobacter spp. (3%), Staphylococcus epidermitis (3%), Streptococci spp. (2%), Proteus spp. (1%), Citrobacter spp. (1%), and Serratia spp. (1%). A diverse range of microbes were identified including clinically relevant bacteria, which shows that the urban water cycle is already contaminated with multidrug-resistant microflora of the hospital settings. Macrolide and lincosamide showed the highest resistance followed by penicillin, monobactam, and cephalosporins. blaSHV and blaTEM were prevalent in samples. blaNDM was also found which manifests as a real threat since it causes resistance against carbapenems and colistin, antibiotics reserved as a last resort against infections. Conclusions This study presented the ground reality of antibiotic resistance in Pakistan and how its subsequent spread poses a great threat to the strides made in the field of medicine and public health. Strict regulations regarding antibiotic usage, hospital effluent, and urban water sanitation must be imposed to curb the devastating effects of this increasing phenomenon.202134790487
122550.9414Escherichia coli serogroups in slaughterhouses: Antibiotic susceptibility and molecular typing of isolates. This study aimed to investigate the contamination of carcasses and slaughterhouse environment with Escherichia coli O157:H7 and non-O157 serogroups (O45:H2, O103:H2, O121:H19, O145:H28, O26:H11, O111:H8). For this purpose, a total of 150 samples (30 carcasses, 30 shredding units, 30 knives, 30 slaughterhouse waste water and 30 wall surfaces) were collected from 5 different slaughterhouses in Kayseri, Turkey. The conventional and molecular methods were performed in order to detect Escherichia coli and its serogroups. Of the 150 samples, 55 (36%) were found to be contaminated with E. coli. Among isolates, E. coli serogroup (O157:H7) were detected in 2 (11%) carcass and 2 (11%) wastewater samples. None of the E. coli isolates harbored tested genes (stx1, stx2, eaeA, and hylA). Effective infection control measures and antibiotic stewardship programs should be adopted to limit the spread of multidrug-resistant bacteria. It was also deduced that these isolates resistance to different antibiotics could be hazardous for public health.202235427957
327960.9414A Three-Year Follow-Up Study of Antibiotic and Metal Residues, Antibiotic Resistance and Resistance Genes, Focusing on Kshipra-A River Associated with Holy Religious Mass-Bathing in India: Protocol Paper. BACKGROUND: Antibiotic resistance (ABR) is one of the major health emergencies for global society. Little is known about the ABR of environmental bacteria and therefore it is important to understand ABR reservoirs in the environment and their potential impact on health. METHOD/DESIGN: Quantitative and qualitative data will be collected during a 3-year follow-up study of a river associated with religious mass-bathing in Central India. Surface-water and sediment samples will be collected from seven locations at regular intervals for 3 years during religious mass-bathing and in absence of it to monitor water-quality, antibiotic residues, resistant bacteria, antibiotic resistance genes and metals. Approval has been obtained from the Ethics Committee of R.D. Gardi Medical College, Ujjain, India (No. 2013/07/17-311). RESULTS: The results will address the issue of antibiotic residues and antibiotic resistance with a focus on a river environment in India within a typical socio-behavioural context of religious mass-bathing. It will enhance our understanding about the relationship between antibiotic residue levels, water-quality, heavy metals and antibiotic resistance patterns in Escherichia coli isolated from river-water and sediment, and seasonal differences that are associated with religious mass-bathing. We will also document, identify and clarify the genetic differences/similarities relating to phenotypic antibiotic resistance in bacteria in rivers during religious mass-bathing or during periods when there is no mass-bathing.201728555050
106370.9413Enterobacteriaceae resistant to third-generation cephalosporins and quinolones in fresh culinary herbs imported from Southeast Asia. Since multidrug resistant bacteria are frequently reported from Southeast Asia, our study focused on the occurrence of ESBL-producing Enterobacteriaceae in fresh imported herbs from Thailand, Vietnam and Malaysia. Samples were collected from fresh culinary herbs imported from Southeast Asia in which ESBL-suspected isolates were obtained by selective culturing. Analysis included identification by MALDI-TOF mass spectrometry, susceptibility testing, XbaI-PFGE, microarray, PCR and sequencing of specific ESBL genes, PCR based replicon typing (PBRT) of plasmids and Southern blot hybridization. In addition, the quinolone resistance genotype was characterized by screening for plasmid mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance determining region (QRDR) of gyrA and parC. The study encompassed fifty samples of ten batches of culinary herbs (5 samples per batch) comprising nine different herb variants. The herbs originated from Thailand (Water morning glory, Acacia and Betel leaf), Vietnam (Parsley, Asian pennywort, Houttuynia leaf and Mint) and Malaysia (Holy basil and Parsley). By selective culturing 21 cefotaxime resistant Enterobacteriaceae were retrieved. Array analysis revealed 18 isolates with ESBL genes and one isolate with solely non-ESBL beta-lactamase genes. Mutations in the ampC promoter region were determined in two isolates with PCR and sequencing. The isolates were identified as Klebsiella pneumoniae (n=9), Escherichia coli (n=6), Enterobacter cloacae complex (n=5) and Enterobacter spp. (n=1). All isolates tested were multidrug resistant. Variants of CTX-M enzymes were predominantly found followed by SHV enzymes. PMQR genes (including aac(6')-1b-cr, qnrB and qnrS) were also frequently detected. In almost all cases ESBL and quinolone resistance genes were located on the same plasmid. Imported fresh culinary herbs from Southeast Asia are a potential source for contamination of food with multidrug resistant bacteria. Because these herbs are consumed without appropriate heating, transfer to human bacteria cannot be excluded.201424607424
122380.9413Characterization of Escherichia coli virulence genes, pathotypes and antibiotic resistance properties in diarrheic calves in Iran. BACKGROUND: Calf diarrhea is a major economic concern in bovine industry all around the world. This study was carried out in order to investigate distribution of virulence genes, pathotypes, serogroups and antibiotic resistance properties of Escherichia coli isolated from diarrheic calves. RESULTS: Totally, 76.45% of 824 diarrheic fecal samples collected from Isfahan, Chaharmahal, Fars and Khuzestan provinces, Iran were positive for E. coli and all of them were also positive for cnf2, hlyA, cdtIII, f17c, lt, st, stx1, eae, ehly, stx2 and cnf1 virulence genes. Chaharmahal had the highest prevalence of STEC (84.61%), while Isfahan had the lowest (71.95%). E. coli serogroups had the highest frequency in 1-7 days old calves and winter season. Distribution of ETEC, EHEC, AEEC and NTEC pathotypes among E. coli isolates were 28.41%, 5.07%, 29.52% and 3.49%, respectively. Statistical analyses were significant for presence of bacteria between various seasons and ages. All isolates had the high resistance to penicillin (100%), streptomycin (98.25%) and tetracycline (98.09%) antibiotics. The most commonly detected resistance genes were aadA1, sul1, aac[3]-IV, CITM, and dfrA1. The most prevalent serogroup among STEC was O26. CONCLUSIONS: Our findings should raise awareness about antibiotic resistance in diarrheic calves in Iran. Clinicians should exercise caution when prescribing antibiotics.201425052999
145890.9413Molecular characterization of extended spectrum β -lactamases enterobacteriaceae causing lower urinary tract infection among pediatric population. BACKGROUND: The β-lactam antibiotics have traditionally been the main treatment of Enterobacteriaceae infections, nonetheless, the emergence of species producing β- Lactamases has rendered this class of antibiotics largely ineffective. There are no published data on etiology of urinary tract infections (UTI) and antimicrobial resistance profile of uropathogens among children in Qatar. The aim of this study is to determine the phenotypic and genotypic profiles of antimicrobial resistant Enterobacteriaceae among children with UTI in Qatar. METHODS: Bacteria were isolated from 727 urine positive cultures, collected from children with UTI between February and June 2017 at the Pediatric Emergency Center, Doha, Qatar. Isolated bacteria were tested for antibiotic susceptibility against sixteen clinically relevant antibiotics using phoenix and Double Disc Synergy Test (DDST) for confirmation of extended-spectrum beta-lactamase (ESBL) production. Existence of genes encoding ESBL production were identified using polymerase chain reaction (PCR). Statistical analysis was done using non-parametric Kappa statistics, Pearson chi-square test and Jacquard's coefficient. RESULTS: 201 (31.7%) of samples were confirmed as Extended Spectrum β -Lactamases (ESBL) Producing Enterobacteriaceae. The most dominant pathogen was E. coli 166 (83%) followed by K. pneumoniae 22 (11%). Resistance was mostly encoded by (bla) CTX-M (59%) genes, primarily (bla) CTX-MG1 (89.2%) followed by (bla) CTX-MG9 (7.7%). 37% of isolated bacteria were harboring multiple (bla) genes (2 genes or more). E. coli isolates were categorized into 11 clusters, while K. pneoumoniae were grouped into five clonal clusters according to the presence and absence of seven genes namely (bla) TEM, (bla) SHV, (bla) CTX-MG1, (bla) CTX-MG2, (bla) CTX-MG8 (bla) CTX-MG9,(bla) CTX-MG25. CONCLUSIONS: Our data indicates an escalated problem of ESBL in pediatrics with UTI, which mandates implementation of regulatory programs to reduce the spread of ESBL producing Enterobacteriaceae in the community. The use of cephalosporins, aminoglycosides (gentamicin) and trimethoprim/sulfamethoxazole is compromised in Qatar among pediatric population with UTI, leaving carbapenems and amikacin as the therapeutic option for severe infections caused by ESBL producers.201830069306
2646100.9413Detection of Antimicrobial Resistance Genes in Escherichia coli Isolated from Black Howler Monkeys (Alouatta pigra) and Domestic Animals in Fragmented Rain-Forest Areas in Tabasco, Mexico. The appearance and spread of antimicrobial resistance (AMR) in bacteria in natural environments and wildlife are related to agricultural and livestock activities and are a global health and conservation problem. We assessed the presence of AMR genes in Escherichia coli isolated from black howler monkeys (Alouatta pigra), sheep (Ovis aries), cattle (Bos taurus), and horses (Equus caballus) from a highly fragmented forest in southern Mexico. Fresh fecal samples were collected using swabs, seeded on eosin-methylene blue agar, and E. coli colonies identified by PCR; multiplex-PCR was performed on E. coli DNA for the detection of 10 AMR genes from four families (sulfonamides, tetracycline, β-lactamase, and chloramphenicol). We detected E. coli in 94% (48/51) of fecal samples, of which 33% (16/48) tested positive for at least one AMR gene. We detected AMR genes in at least one individual from each sampled animal species, with the most prevalent genes being tet(B) 18% (9/48), sul2 14% (7/48), sul1, and blaTEM 12% (6/48). Sheep samples contained AMR genes from the four families of antibiotics detected in this study and 50% (5/10) tested positive for the presence of at least one gene. A total of 12% (2/16) of fecal samples from black howler monkeys tested positive for AMR genes. The presence of AMR genes in A. pigra and domestic animals has not been reported in the Balancán area of Tabasco, Mexico. Transmission of AMR bacteria from domestic animals to monkeys is rare; however, this is a potential health risk for wildlife and species conservation.202032402234
1237110.9412Characterization 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
2170120.9411Drug resistance in bacteria isolated from patients presenting with wounds at a non-profit Surgical Center in Phnom Penh, Cambodia from 2011-2013. BACKGROUND: Emerging antibiotic resistance amongst clinically significant bacteria is a public health issue of increasing significance worldwide, but it is relatively uncharacterized in Cambodia. In this study we performed standard bacterial cultures on samples from wounds at a Non-Governmental-Organization (NGO) Hospital in Phnom Penh, Cambodia. Testing was performed to elucidate pathogenic bacteria causing wound infections and the antibiotic resistance profiles of bacterial isolates. All testing was performed at the Naval Medical Research Unit, No.2 (NAMRU-2) main laboratory in Phnom Penh, Cambodia. METHODS: Between 2011-2013, a total of 251 specimens were collected from patients at the NGO hospital and analyzed for bacterial infection by standard bacterial cultures techniques. Specimens were all from wounds and anonymous. No specific clinical information accompanied the submitted specimens. Antibiotic susceptibility testing, and phenotypic testing for extended-spectrum beta-lactamase (ESBL) were performed and reported based on CLSI guidelines. Further genetic testing for CTX-M, TEM and SHV ESBLs was accomplished using PCR. RESULTS: One-hundred and seventy-six specimens were positive following bacterial culture (70 %). Staphlycoccus aureus was the most frequently isolated bacteria. Antibiotic drug resistance testing revealed that 52.5 % of Staphlycoccus aureus isolates were oxacillin resistant. For Escherichia coli isolates, 63.9 % were ciprofloxacin and levofloxacin resistant and 96 % were ESBL producers. Resistance to meropenem and imipenem was observed in one of three Acinetobacter spp isolates. CONCLUSIONS: This study is the first of its kind detailing the antibiotic resistance profiles of pathogenic bacteria causing wound infections at a single surgical hospital in Cambodia. The reported findings of this study demonstrate significant antibiotic resistance in bacteria from injured patients and should serve to guide treatment modalities in Cambodia.201528883936
1238130.9411Lineages, Virulence Gene Associated and Integrons among Extended Spectrum β-Lactamase (ESBL) and CMY-2 Producing Enterobacteriaceae from Bovine Mastitis, in Tunisia. Extended Spectrum Beta-Lactamase (ESBL) Enterobacteriaceae are becoming widespread enzymes in food-producing animals worldwide. Escherichia coli and Klebseilla pneumoniae are two of the most significant pathogens causing mastitis. Our study focused on the characterization of the genetic support of ESBL/pAmpC and antibiotic resistance mechanisms in cefotaxime-resistant (CTXR) and susceptible (CTXS) Enterobacteriaceae isolates, recovered from bovine mastitis in Tunisia, as well as the analyses of their clonal lineage and virulence-associated genes. The study was carried out on 17 ESBL/pAmpC E. coli and K. pneumoniae and 50 CTXS E. coli. Detection of resistance genes and clonal diversity was performed by PCR amplification and sequencing. The following β-lactamase genes were detected: blaCTX-M-15 (n = 6), blaCTX-M-15 + blaOXA-1 (2), bla CTX-M-15 + blaOXA-1 + blaTEM-1b (2), blaCTX-M-15 + blaTEM-1b (4), blaCMY-2 (3). The MLST showed the following STs: ST405 (n = 4 strains); ST58 (n = 3); ST155 (n = 3); ST471 (n = 2); and ST101 (n = 2). ST399 (n = 1) and ST617 (n = 1) were identified in p(AmpC) E. coli producer strains. The phylogroups A and B1 were the most detected ones, followed by the pathogenic phylogroup B2 that harbored the shigatoxin genes stx1/stx2, associated with the cnf, fimA, and aer virulence factors. The qnrA/qnrB, aac(6′)-Ib-cr genes and integrons class 1 with different gene cassettes were detected amongst these CTXR/S isolated strains. The presence of different genetic lineages, associated with resistance and virulence genes in pathogenic bacteria in dairy farms, may complicate antibiotic therapies and pose a potential risk to public health.202236015067
1346140.9410High prevalence of multidrug resistant Escherichia coli isolated from fresh vegetables sold by selected formal and informal traders in the most densely populated Province of South Africa. Contaminated fresh produce has increasingly been implicated in foodborne disease outbreaks. As microbiological safety surveillance in South Africa is limited, a total of 545 vegetable samples (spinach, tomato, lettuce, cucumber, and green beans) were purchased from retailers, street traders, trolley vendors and farmers' markets. Escherichia coli, coliforms and Enterobacteriaceae were enumerated and the prevalence of Escherichia coli, Salmonella spp. and Listeria monocytogenes determined. E. coli isolates were characterized phenotypically (antibiotic resistance) and genotypically (diarrheagenic virulence genes). Coliforms, E. coli and Enterobacteriaceae counts were mostly not significantly different between formal and informal markets, with exceptions noted on occasion. When compared to international standards, 90% to 98% tomatoes, 70% to 94% spinach, 82% cucumbers, 93% lettuce, and 80% green bean samples, had satisfactory (≤ 100 CFU/g) E. coli counts. Of the 545 vegetable samples analyzed, 14.86% (n = 81) harbored E. coli, predominantly from leafy green vegetables. Virulence genes (lt, st, bfpA, eagg, eaeA, stx1, stx2, and ipaH) were not detected in the E. coli isolates (n = 67) characterized, however 40.30% were multidrug-resistant. Resistance to aminoglycosides (neomycin, 73.13%; gentamycin, < 10%), penicillins (ampicillin, 38.81%; amoxicillin, 41.79%; augmentin, < 10%), sulfonamides (cotrimoxazole, 22.39%), tetracycline (19.4%), chloramphenicol (11.94%), cephalosporins (cefepime, 34.33%), and carbapenemases (imipenem, < 10%) were observed. This study highlights the need for continued surveillance of multidrug resistant foodborne pathogens in fresh produce retailed formally and informally for potential consumer health risks. PRACTICAL APPLICATION: The results indicate that the microbiological quality of different vegetables were similar per product type, regardless of being purchased from formal retailers or informal street traders, trolley vendors or farmers' markets. Although no pathogenic bacteria (diarrheagenic E. coli, Salmonella spp. or L. monocytogenes) were isolated, high levels of multidrug-resistance was observed in the generic E. coli isolates. These findings highlight the importance of microbiological quality surveillance of fresh produce in formal and informal markets, as these products can be a reservoir of multidrug resistant bacteria harboring antibiotic resistance and virulence genes, potentially impacting human health.202133294974
2941150.9410Uncovering hidden threats: prevalence, antibiotic resistance and virulence gene profiles of Escherichia coli strains isolated from Testudines and their aquatic habitats. BACKGROUND: The gut microbiota of Testudines is fundamental to their digestion and overall health, yet remains a poorly investigated area in their biology, particularly in wild freshwater turtle (terrapins) and tortoise populations within South Africa. This study investigated the occurrence, diversity, virulence genes and antibiotic resistance of Escherichia coli isolated from Testudine gut microbiota and sediments at Timbavati Private Nature Reserve, South Africa. METHODS AND RESULTS: Cloacal swab samples were collected from 36 wild Testudines and 20 sediment samples from temporary and permanent water bodies. Presumed E. coli isolates were confirmed by polymerase chain reaction (PCR) targeting the β-D glucuronidase (uidA) gene and further validated through 16 S rRNA gene sequencing. Phenotypic antibiotic resistance was evaluated with the Kirby-Bauer method, whilst resistance and virulence genes were identified using PCR assays. E. coli was detected in 54 (62%) of 87 isolates (23 Testudines and 31 sediments), confirmed by uidA PCR assay. Detected virulence genes included eaeA (42%), virF (22%), stx1 (16%), and stx2 (3%), and isolates exhibited resistance to erythromycin (53%), cephalothin (48%), and spectinomycin (40%). Resistance genes such as mcr-4 (70%), bla(SHV) (46%), bla(TEM) (64%), mcr-1 (42%), qnrA (16%), mcr-2 (22%), qnrD (11%), and tetW (2%) were also detected. CONCLUSIONS: This study demonstrates that wild Testudines harbour E. coli in their gut and that it also occurs in their surrounding environment, with notable antibiotic resistance and virulence potential. The findings underscore the complexity of host-microbial interactions and the influence of environmental and host factors on microbial diversity, informing potential conservation and health management strategies for these reptilian species.202540751752
1233160.9410Prevalence, 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
2129170.9409Screening of antibiotic resistance genes in pathogenic bacteria isolated from tiny freshwater shrimp (Macrobrachium lanchesteri) and "Kung Ten", the uncooked Thai food. OBJECTIVE: This study aimed to isolate and identify of pathogenic bacteria in tiny freshwater shrimp (Macrobrachium lanchesteri) and in Kung Ten, which is an unusual Thai cuisine that eaten alive shrimp directly. Antimicrobial susceptibility test and identification of antibiotic resistance genes for isolated bacteria were conducted. MATERIALS AND METHODS: Eighty of fresh shrimp samples and forty of Kung Ten salads were collected from four fresh markets, which were located in Bangkok and Nonthaburi province (N = 120). The isolation, identification, and antimicrobial susceptibility test of pathogenic bacteria were done following the Clinical and Laboratory Standards Institute guidelines. Antibiotic-resistant bacteria were screened for β-lactamase relating genes, such as AmpC (MOX and ACC genes), bla (CTX-M), and Int1 genes. RESULTS: The number of bacterial isolates in tiny freshwater shrimp and Kung Ten salad was 136 and 65, respectively. Aeromonas caviae, A. hydrophilla, Proteus penneri, Proteus vulgaris, and Klebsiella pneumoniae were commonly found. Ampicillin, amoxicillin/clavulanic, cefuroxime, tetracycline, and trimethoprim/sulfamethoxazole resistance were observed, and common antibiotic-resistant bacteria were A. caviae, P. vulgaris, Enterobacter Aerogenes, and K. pneumoniae. A. caviae, P. penneri, K. Pneumoniae, and A. hydrophilla were positive for MOX gene; bla (CTX-M), and Int1 genes; ACC and Int1 genes; and ACC gene, respectively. CONCLUSION: Raw or uncooked shrimps in Kung Ten salad may a risk in foodborne diseases due to positive for pathogenic bacterial isolates. However, hygienic control on food preparation is difficult to apply because of the difficulty of changing in local Thai food behavior.202032219114
971180.9409Paper currency harbours antibiotic-resistant coliform bacteria and integron integrase. AIMS: This study was designed to analyse the prevalence of class 1 and class 2 integron integrase genes among antibiotic-resistant coliform bacteria isolated from paper currency circulating in Pakistan. METHODS AND RESULTS: A total of 500 individual currency notes were collected from different food vending sites at Lahore, Pakistan. Bacterial population were identified by biochemical and PCR techniques. Antimicrobial susceptibility testing was performed by disc diffusion assay. The highest bacterial population on currency was found from street vendors and butcher shops. Escherichia coli was found to be the most prevalent coliform bacteria followed by Klebsiella sp. and Enterobacter sp. PCR amplification of antimicrobial resistance gene showed the presence of ampC, bla(TEM) , bla(NDM-1) , qnrA, tet(A) and tet(B) genes among coliform isolates. A total of 47 integron integrase bearing strains of coliform bacteria were analysed. Sequence analysis showed the presence of dfrA1-aadA1, dfrA1, dfrA5, dfrA7, aadA1, aadA4 cassette arrays in class 1 integron and dfrA1-sat2-aadA1 in class 2 integrase genes. CONCLUSION: Circulating currency was heavily contaminated with antimicrobial-resistant coliform bacteria bearing class 1 and class 2 integron integrase genes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study describes a potential threat of severe bacterial infections due to improper hand hygiene and community sanitation when dealing with the currency notes.202132966644
1434190.9408Molecular characterization of carbapenemases production among environmental Gram-negative isolates at Addis Ababa, Ethiopia: first detection of NDM Producers in hospital environments. INTRODUCTION: The Gram-Negative bacteria, particularly carbapenem-resistant strains (CR-GNB), pose a global health threat due to high morbidity and mortality. Detecting carbapenemase-encoding genes is essential for understanding their spread in hospital environments. This study investigated environmental colonization by CR-GNB in Ethiopian hospitals, including genetic characterization of resistance genes. METHODOLOGY: A cross-sectional study analyzed 103 environmental GNB isolates collected from inanimate surfaces at Tikur Anbessa Specialized Hospital (TASH) and ALERT Hospital (June-September 2021). Conventional microbiological methods identified the isolates, and antimicrobial susceptibility was tested using the Kirby-Bauer disk diffusion method. Carbapenemase production was screened using the Modified Hodge test (MHT) and combined disk test (CDT). Resistance genes (blaKPC, blaNDM, blaOXA-48) were detected via PCR in isolates with reduced meropenem susceptibility. RESULTS: The predominant GNB were Acinetobacter baumannii (47%), Pseudomonas aeruginosa (33%), and E. coli (12%). Among 103 isolates, 62% showed reduced meropenem susceptibility. The most common CR-GNB was Acinetobacter baumannii (37.5%), followed by E. coli (18.8%) and Klebsiella pneumoniae (12.5%). Carbapenemase production was detected in 41.7% of isolates via PCR, with blaNDM being the most common (43 isolates). Linens (26.4%) and beds (21.4%) had the highest contamination rates. Most carbapenemase-producing isolates were multidrug-resistant (MDR). CONCLUSIONS: The presence of blaNDM and blaKPC genes highlights hospital surfaces as reservoirs for resistance genes, contributing to healthcare-associated infections. Routine surveillance and early detection of carbapenemase producers are crucial for infection control and antimicrobial resistance management.202540305531