Escherichia coli tetracycline efflux determinants in relation to tetracycline residues in chicken. - Related Documents




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232301.0000Escherichia coli tetracycline efflux determinants in relation to tetracycline residues in chicken. OBJECTIVE: To screen for Escherichia coli (E. coli) resistant to tetracycline, followed by identification of tet efflux genes by polymerase chain reaction (PCR). In addition, detection of tetracycline residues in chicken livers and kidneys were conducted using high performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS). METHODS: Strains of E. coli were isolated from samples of chicken colon and screened for tetracycline resistance. Tetracycline genes conferring resistance (Tc(r)) were detected by polymerase chain reaction (PCR). Most of the isolates were resistant to tetracycline (97.9%). RESULTS: PCR analysis indicated that Tc(r)E. coli R-plasmids contained tet(A), tet(B) and a combination of both efflux genes. None of the isolates contained other efflux tet genes tet (C, D, E and Y). High performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS), a sensitive technique, was used to detect residues of chlortetracycline (CTC), oxytetracycline (OTC), doxycycline (DC) in chicken livers and kidneys. The samples containing tetracycline residues were at 0.13-0.65 pg/μL levels. CONCLUSIONS: Tetracycline and other antibiotics are commonly used in the poultry and meat production industry for prevention of microbial infections. Multiple antibiotic resistant bacteria in Oman have increased to alarming levels, threatening public health, domestic and may have adverse effect on environment.201323827150
270310.9998Detection of multi-drug resistant (MDR) Escherichia coli and tet gene prevalence at a pig farm in Kupang , Indonesia. OBJECTIVE: The purpose of this study was to detect the incidence of multi-drug resistant (MDR) and the spread of tet genes that encode tetracycline (TE) resistance in E. coli in pig farms in the city of Kupang, Indonesia. MATERIALS AND METHODS: Samples of pig feces have been obtained from 96 pig farms in Kupang city, Indonesia. Escherichia coli bacteria were isolated and identified morphologically and biochemically, and finally confirmed by the API test. The disk diffusion method has been used to observe the antibiotic sensitivity effects and has been followed by observing resistant genes encoding TE resistance using the multiplex polymerase chain reaction (m-PCR) method to detect the presence of tet genes such as tet (A), tet (B), tet (C), tet (D), and tet (E), respectively. RESULTS: A total of 82 (85.4%) of E. coli isolates have been found in all pig feces samples obtained from 96 pig farms in Kupang city. This study has shown a high level of antibiotic resistance dominated by erythromycin (85.4%) and cephalothin (58.5%) and followed by several other antibiotics with a percentage below 34.1%. The prevalence of MDR E. coli was 57.3% by showing 39 different patterns. The most common pattern was showed by the Cephalothin-Colistin-Erythromycin pattern. The resistance of E. coli to TE appears to be related to the presence of tet (A) and tet (E) genes. CONCLUSION: This study has encouraged the need for public awareness (farmers) of the wise use of antibiotics in preventing the spread of resistant bacteria that can cause health problems in animals and humans.201831453148
554420.9998Assessing the Effect of Oxytetracycline on the Selection of Resistant Escherichia coli in Treated and Untreated Broiler Chickens. Oxytetracycline (OTC) is administered in the poultry industry for the treatment of digestive and respiratory diseases. The use of OTC may contribute to the selection of resistant bacteria in the gastrointestinal tract of birds or in the environment. To determine the effect of OTC on the selection of resistant Escherichia coli strains post-treatment, bacteria were isolated from droppings and litter sampled from untreated and treated birds. Bacterial susceptibility to tetracyclines was determined by the Kirby-Bauer test. A total of 187 resistant isolates were analyzed for the presence of tet(A), (B), (C), (D), (E), and (M) genes by PCR. Fifty-four strains were analyzed by PFGE for subtyping. The proportion of tetracycline-resistant E. coli strains isolated was 42.88%. The susceptibility of the strains was treatment-dependent. A high clonal diversity was observed, with the tet(A) gene being the most prevalent, followed by tet(C). Even at therapeutic doses, there is selection pressure on resistant E. coli strains. The most prevalent resistance genes were tet(A) and tet(C), which could suggest that one of the main mechanisms of resistance of E. coli to tetracyclines is through active efflux pumps.202338136686
290830.9998Detection of tetracycline and macrolide resistance determinants in Enterococci of animal and environmental origin using multiplex PCR. An occurrence of resistance to tetracycline (TET) and erythromycin (ERY) was ascertained in 82 isolates of Enterococcus spp. of animal and environmental origin. Using E test, 33 isolates were resistant to TET and three isolates to ERY. Using polymerase chain reaction (PCR; single and multiplex), the TET determinants tet(M) and tet(L) were detected in 35 and 13 isolates, respectively. Twelve isolates carried both tet(M) and tet(L) genes. Eight isolates possessed ermB gene associated with ERY resistance. Multiplex PCR was shown to be a suitable method for simultaneous determination of all three resistance determinants that occurred most frequently in bacteria isolated from poultry. This study also demonstrates that gastrointestinal tract of broilers may be a reservoir of enterococci with acquired resistance to both TET and ERY that can be transferred to humans via food chain.201121656006
286840.9998Detection and Analysis of Drug and Disinfectant Resistance Genes in the Sewage of a Center for Disease Control and Prevention. PURPOSE: Sewage is a significant reservoir for drug and disinfectant resistance genes and a medium for dissemination. This study aimed to evaluate the presence of drug and disinfectant resistance genes in the sewage of a Center for Disease Control and Prevention (CDC) and to assess the risks of their dissemination. METHODS: Sewage from a CDC in Hangzhou was collected, filtered, and enriched, and its microorganisms were cultured. The isolated bacteria were identified, and the minimum inhibitory concentration (MIC) was determined. The drug and disinfectant resistance genes in the sewage and bacteria were detected through polymerase chain reaction amplification. RESULTS: Three kinds of bacteria were isolated from the sewage sample. The MIC for Sphingomonas and Staphylococcus xylosus against chlorine-containing disinfectants was 250 mg/L, whereas the MIC for Bacillus firmus was 500 mg/L. The β-lactam resistance gene TEM and the disinfectant resistance gene qacA were positive in the bacteria, whereas the β-lactam resistance genes TEM, SHV, and VIM-1, the tetracycline resistance gene tetM, the aminoglycoside resistance genes aac(6')/aph(2') and aph3'-III, and the disinfectant resistance genes qacA, qacE, and qacEΔ1 were positive in the sewage. CONCLUSION: Drug and disinfectant resistance genes were found in the sewage of a CDC and were associated with bacteria. Thus, optimizing the monitoring and treatment of sewage is crucial.202540303605
285450.9998Occurrence of antibiotic resistance genes in culturable bacteria isolated from Turkish trout farms and their local aquatic environment. Antibiotic resistance and presence of the resistance genes were investigated in the bacteria isolated from water, sediment, and fish in trout farms. A total of 9 bacterial species, particularly Escherichia coli, were isolated from the water and sediment samples, and 12 species were isolated from fish. The antimicrobial test indicated the highest resistance against sulfamethoxazole and ampicillin in coliform bacteria, and against sulfamethoxazole, imipenem, and aztreonam in known pathogenic bacteria isolated from fish. The most effective antibiotics were rifampicin, chloramphenicol, and tetracycline. The multiple antibiotic resistance index was above the critical limit for almost all of the bacteria isolated. The most common antibiotic resistance gene was ampC, followed by tetA, sul2, blaCTX-M1, and blaTEM in the coliform bacteria. At least one resistance gene was found in 70.8% of the bacteria, and 66.6% of the bacteria had 2 or more resistance genes. Approximately 36.54% of the bacteria that contain plasmids were able to transfer them to other bacteria. The plasmid-mediated transferable resistance genes were ampC, blaCTX-M1, tetA, sul2, and blaTEM. These results indicate that the aquatic environment could play an important role in the development of antibiotic resistance and the dissemination of resistance genes among bacteria.201525993887
292260.9998Tetracycline-resistance genes in gram-negative isolates from estuarine waters. AIMS: To investigate the diversity and dissemination of tetracycline resistance genes in isolates from estuarine waters. METHODS AND RESULTS: Forty-two out of 164 multi-resistant isolates previously obtained were resistant or less-susceptible to tetracycline, as evaluated by the disc diffusion method. Minimal inhibitory concentration for resistant bacteria ranged from 16 to 256 mg l(-1). Screening of tet genes by polymerase chain reaction showed that 88% of the isolates carried at least one of the genes tested, namely tet(A) (present in 13 isolates), tet(B) (present in 13 isolates), tet(C) (present in 3 isolates), tet(D) (present in 1 isolate), tet(E) (present in 6 isolates) and tet(M) (present in 1 isolate). One isolate carried tet(A) and tet(M). To our knowledge, this study presents the first description of a tet(D) gene in Morganella morganii. Hybridization revealed that tet genes were plasmid-located in 31% of the isolates. Those isolates were included as donors in conjugation experiments and 38% transferred tetracycline resistance. CONCLUSIONS: A considerable diversity of tet genes was detected in the estuary. Frequently, these genes were associated with plasmids and could be transferred to Escherichia coli. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented provide further evidence of the role played by estuarine reservoirs in antibiotic resistance maintenance and dissemination.200819120920
268870.9998Intestinal and Extraintestinal Pathotypes of Escherichia coli Are Prevalent in Food Prepared and Marketed on the Streets from the Central Zone of Mexico and Exhibit a Differential Phenotype of Resistance Against Antibiotics. Background/Objectives: Antibiotic resistance is a serious public health problem threatening the treatment of infectious diseases caused by Escherichia coli, the main source of food contamination and responsible for many infectious diseases with high indices of AR profiles. Our objective was to study the presence of Escherichia coli in foods that are distributed and prepared on the street, characterizing its sensitivity profile and resistance to antibiotic drugs commonly prescribed in this geographical area. Methods: Standard procedures were performed to identify and isolate E. coli colonies from food samples collected during a three-year study. Susceptibility assays were conducted to determine the antibiotic resistance profile, and Colony PCR assays were performed to determine the pathogenic and antibiotic resistance genes. Results: A total of 189 food samples were collected, and 100% of the samples were positive for E. coli, with higher percentages of contamination for vegetables and fruits. ETEC (lt) and UPEC (vat, cnf1, hylA) genes were identified in 100% of the samples and DAEC (afa) in 27%. E. coli exhibited high percentages of resistance against ampicillin and amoxicillin/clavulanic acid (100%) and cephalexin (45%). The most effective antibiotics were tetracycline, TMP-SMX, polymyxin, and quinolones. The AR genes tetA, sul1, catA1, strA, qnrS, and floR were identified among the samples. Conclusions: Food prepared and marketed on the streets seriously threatens human health. Ampicillin and amoxicillin/clavulanic acid should not be used to treat infections caused by the multidrug-resistant ETEC and UPEC identified in this area. To our knowledge, this is the first study that explores the status of AR in this geographical area.202540298585
290480.9998The maintenance in the oral cavity of children of tetracycline-resistant bacteria and the genes encoding such resistance. OBJECTIVES: To investigate the maintenance of tetracycline-resistant oral bacteria and the genes encoding tetracycline resistance in these bacteria in children (aged 4--6 years) over a period of 12 months. METHODS: Plaque and saliva samples were taken from 26 children. Tetracycline-resistant bacteria were isolated and identified. The types of resistance genes and their genetic locations were also determined. RESULTS: Fifteen out of 18 children harboured tetracycline-resistant (defined as having a MIC>or=8 mg/L) oral bacteria at all three time points. The median percentage of tetracycline-resistant bacteria at 0, 6 and 12 months was 1.37, 1.37 and 0.85%, respectively; these were not significantly different. The MIC(50) of the group was 64 mg/L at all three time points compared with the MIC(90), which was 64 mg/L at 0 months, and 128 mg/L at 6 and 12 months. The most prevalent resistant species were streptococci (68%), which were isolated at all three time points in 13 children. The most prevalent gene encoding tetracycline resistance was tet(M) and this was found in different species at all three time points. For the first time, tet(32) was found in Streptococcus parasanguinis and Eubacterium saburreum. PCR and Southern-blot analysis (on isolates from three of the children) showed that the tet(M) gene was located on a Tn916-like element and could be detected at all three time points, in four different genera, Streptococcus, Granulicatella, Veillonella and Neisseria. CONCLUSIONS: The results of this study show that tetracycline-resistant bacteria and tet(M) are maintained within the indigenous oral microbiota of children, even though they are unlikely to have been directly exposed to tetracycline.200516027144
592190.9998Prevalence of tetracycline resistance genes in oral bacteria. Tetracycline is a broad-spectrum antibiotic used in humans, animals, and aquaculture; therefore, many bacteria from different ecosystems are exposed to this antibiotic. In order to determine the genetic basis for resistance to tetracycline in bacteria from the oral cavity, saliva and dental plaque samples were obtained from 20 healthy adults who had not taken antibiotics during the previous 3 months. The samples were screened for the presence of bacteria resistant to tetracycline, and the tetracycline resistance genes in these isolates were identified by multiplex PCR and DNA sequencing. Tetracycline-resistant bacteria constituted an average of 11% of the total cultivable oral microflora. A representative 105 tetracycline-resistant isolates from the 20 samples were investigated; most of the isolates carried tetracycline resistance genes encoding a ribosomal protection protein. The most common tet gene identified was tet(M), which was found in 79% of all the isolates. The second most common gene identified was tet(W), which was found in 21% of all the isolates, followed by tet(O) and tet(Q) (10.5 and 9.5% of the isolates, respectively) and then tet(S) (2.8% of the isolates). Tetracycline resistance genes encoding an efflux protein were detected in 4.8% of all the tetracycline-resistant isolates; 2.8% of the isolates had tet(L) and 1% carried tet(A) and tet(K) each. The results have shown that a variety of tetracycline resistance genes are present in the oral microflora of healthy adults. This is the first report of tet(W) in oral bacteria and the first report to show that tet(O), tet(Q), tet(A), and tet(S) can be found in some oral species.200312604515
5917100.9998Heavy metal co-resistance with antibiotics amongst bacteria isolates from an open dumpsite soil. Heavy metal co-resistance with antibiotics appears to be synergistic in bacterial isolates via similar mechanisms. This synergy has the potential to amplify antibiotics resistance genes in the environment which can be transferred into clinical settings. The aim of this study was to assess the co-resistance of heavy metals with antibiotics in bacteria from dumpsite in addition to physicochemical analysis. Sample collection, physicochemical analysis, and enumeration of total heterotrophic bacteria counts (THBC) were all carried out using standard existing protocols. Identified bacteria isolates were subjected to antibiotics sensitivity test using the Kirby Bauer disc diffusion technique and the resulting multidrug resistant (MDR) isolates were subjected to heavy metal tolerance test using agar dilution technique with increasing concentrations (50, 100, 150, 200 and to 250 μg/ml) of our study heavy metals. THBC ranged from 6.68 to 7.92 × 10(5) cfu/g. Out of the 20 isolates subjected to antibiotics sensitivity, 50% (n = 10) showed multiple drug resistance and these were B. subtilis, B. cereus, C. freundii, P. aeruginosa, Enterobacter sp, and E. coli (n = 5). At the lowest concentration (50 μg/ml), all the MDR isolates tolerated all the heavy metals, but at 250 μg/ml, apart from cadmium and lead, all test isolates were 100% sensitive to chromium, vanadium and cobalt. The control isolate was only resistant to cobalt and chromium at 50 μg/ml, but sensitive to other heavy metals at all concentrations The level of co-resistance shown by these isolates is a call for concern.202336820045
2921110.9998Diversity of tetracycline resistance genes in bacteria from aquaculture sources in Australia. AIMS: To determine the genetic determinants responsible for tetracycline resistance in oxytetracycline resistant bacteria from aquaculture sources in Australia. METHODS AND RESULTS: Twenty of 104 (19%) isolates tested were resistant to oxytetracycline (MIC > or = 16 microg ml(-1)). Using polymerase chain reaction (PCR) amplification, one or more tet genes were detected in 15/20 (75%) isolates tested, but none were found in 5/20 (25%). tetM (50%) was the most common determinant, followed by tetE (45%), tetA (35%) and tetD (15%). Five of 12 oxytetracycline resistant isolates studied were able to transfer their R-plasmid to Escherichia coli recipients of chicken, pig and human origin. tetA, tetD and tetM were found to be transferred while tetE was not transferred. Southern hybridization and PCR were used to confirm transfer of determinants. CONCLUSIONS: Bacterial isolates from aquaculture sources in Australia harbour a variety of tetracycline resistance genes, which can be transferred to other bacteria of different origin. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteria from aquaculture sources in Australia contribute to the resistance gene pool reservoir. The in vitro transfer of tetracycline R-plasmid from aquatic bacteria to E. coli isolates from various sources is an indication of the potential public health risk associated with these resistance determinants.200717953612
2862120.9997Regulation Transcriptional of Antibiotic Resistance Genes (ARGs) in Bacteria Isolated from WWTP. The incidence of antibiotics and transcriptional regulation of ARGs in isolated bacteria from wastewater needs to be explored. By HPLC, in samples of untreated wastewater, ampicillin (49.74 ± 5.70 µg/mL), chloramphenicol (0.60 ± 0.03 µg/mL), tylosin (72.95 ± 2.03 µg/mL), and oxytetracycline (0.22 ± 0.01 µg/mL) was determined. Through metagenomic analysis identified 58 bacterial species belonging to 9 phyla and at least 14 species have shown resistance to a variety of antibiotics. Twenty-two bacterial isolates were proved to be resistant to fifteen antibiotics of new generation and used in medical research to combat infectious diseases. Fourteen strains were shown to harbor plasmids in size ranges of 2-5 Kb, 6-10 Kb and plasmids with size greater than 10 Kb. By quantitative PCR it was possible to identify genes sul, qnr, cat1, aadA1, and sat-1 gene were shown to be present in gDNA samples from treated and untreated samples of wastewater and by relative expression analysis, differential expression of cat1, ermB, act, and tetA genes was demonstrated in strains that showed identity with Escherichia coli, Bacteroides fragilis, and Salmonella thyphi, and that were stressed with different concentrations of antibiotics. The presence of ARGs in untreated water samples, as well as in bacterial isolates, was indicative that in these habitats there are microorganisms that can resist β-lactams, aminoglycosides, tetracyclines, sulfonamides, and quinolones.202337672120
2869130.9997Antibiotic resistance and antibiotic-resistance genes of Pseudomonas spp. and Escherichia coli isolated from untreated hospital wastewater. Hospitals are considered an important factor in the spread of antibiotic-resistant bacteria (ARBs) and antibiotic-resistance genes (ARGs). The purpose of this research was to characterize the microbial populations in hospital wastewater and investigated the prevalence of β-lactamase, SulІ and QnrS resistance genes. In the first step, culture method was used to isolate Pseudomonas aeruginosa and Escherichia coli. In the next step, accurate identification of isolated bacteria was carried out using the polymerase chain reaction (PCR) method, then the resistance of the bacteria at different concentrations of antibiotics (8-128 μg/mL) was examined. Finally the ARGs were detected using the PCR method. The averages of heterotrophic plate count (HPC) and ARB concentration in wastewater samples were 1.8 × 10(8) and 4.3 × 10(6) CFU/100 mL, respectively. The highest resistance rates were found for sulfamethoxazole and the highest resistance rates in the β-lactamase group were for ceftazidime, while highest sensitivity was for gentamicin and there was no isolate that was sensitive to the studied antibiotics. SulІ and QnrS were the highest and lowest abundance of all ARGs in samples respectively and blaSHV was the highest β-lactam resistance gene. Our results indicated an increase in the resistance of identified bacteria to several antibiotics. So it can be concluded that numerous antibiotic-resistant pathogens and vast numbers of ARGs exist in the human body so that their release from hospitals without effective treatment can cause many dangers to the environment and human health.202134280162
2690140.9997Characterization of Cefotaxime- and Ciprofloxacin-Resistant Commensal Escherichia coli Originating from Belgian Farm Animals Indicates High Antibiotic Resistance Transfer Rates. Food-producing animals represent one of the sources of antibiotic resistant commensal bacteria. There is an increasing awareness that these bacteria might have the potential to transfer their resistance genes to other (pathogenic) bacteria. In this study, 50 commensal Escherichia coli strains originating from food-producing animals and resistant to the "highest priority, critically important antibiotics" cefotaxime and/or ciprofloxacin, were selected for further characterization. For each strain (i) an antibiogram, (ii) the phylogenetic group, (iii) plasmid replicon type, (iv) presence and identification of integrons, and (v) antibiotic resistance transfer ratios were determined. Forty-five of these strains were resistant to 5 or more antibiotics, and 6 strains were resistant to 10 or more antibiotics. Resistance was most common to ampicillin (100%), sulfamethoxazole, ciprofloxacin (82%), trimethoprim, tetracycline (74%), cefotaxime, (70%) and ceftazidime (62%). Phylogenetic groups A (62%) and B1 (26%) were most common, followed by C (8%) and E (4%). In 43 strains, more than 1 replicon type was detected, with FII (88%), FIB (70%), and I1 (48%) being the most encountered types. Forty strains, positive for integrons, all harbored a class I integron and seven of them contained an additional class II integron. No class III integrons were detected. The antibiotic resistance transfer was assessed by liquid mating experiments. The transfer ratio, expressed as the number of transconjugants per recipient, was between 10(-5) and 10(0) for cefotaxime resistance and between 10(-7) and 10(-1) for ciprofloxacin resistance. The results of the current study prove that commensal E. coli in food-production animals can be a source of multiple resistance genes and that these bacteria can easily spread their ciprofloxacin and cefotaxime resistance.201829148895
2322150.9997Antimicrobial Susceptibility of Fresh Produce-Associated Enterobacteriaceae and Enterococci in Oman. Fresh produce bacteria may have phenotypic and/or genotypic antimicrobial resistance traits that may lead to various consequences on the environment and human health. This study evaluated the susceptibility of fresh produce bacteria (banana, cabbage, capsicum, carrots, cucumber, dates, lettuce, mango, papaya, pomegranate, radish, tomato and watermelon) to chlorhexidine and the antibiotic resistance of enterococci. Eighty-eight Enterobacteriaceae bacteria and 31 enterococci were screened for their susceptibility to chlorhexidine using the broth microdilution method. Susceptibility of enterococci to various antibiotics was determined using agar dilution, colorimetric, and Kirby-Bauer disc diffusion methods. Enterococci were more susceptible to chlorhexidine than Enterobacteriaceae indicated by chlorhexidine minimum inhibitory concentration (MIC) of 1 to 8 µg/mL for the former and 1 to 64 µg/mL for the latter. The IntI 1, qacEΔ1, qacE and qacG genes were distributed weakly in three, two, two, and three Enterobacteriaceae isolates, respectively. Enterococci had resistance to chloramphenicol (3%), tetracycline (19%), erythromycin (68%), ciprofloxacin (55%), and vancomycin (10%) while 19% of them were multi-drug resistant. In conclusion, this research detected a low to moderate level of antibiotic resistance in enterococci. Some Enterobacteriaceae bacteria had reduced chlorhexidine MICs that were not 10x less than the recommended concentration (100-200 µg/mL) in food production areas which might challenge the success of the disinfection processes or have clinical implications if the involved bacteria are pathogens. The prevalence of antimicrobial-resistant bacteria in fresh produce should be monitored in the future.202236230161
5922160.9997Incidence of infectious drug resistance among lactose-fermenting bacteria isolated from raw and treated sewage. Raw and treated sewage samples were examined for antibiotic-resistant, lactose-fermenting bacteria. Approximately 1% of the total lactose-fermenting bacteria were multiply resistant. Of these organisms, 50% were capable of transferring all or part of their resistance to a drug-sensitive recipient. Only 43% of those isolated on media containing a single antibiotic were capable of resistance transfer, whereas 57% of those recovered on multiple antibiotic plates transferred resistance. R factors conferring resistance to chloramphenicol, streptomycin, and tetracycline; streptomycin and tetracycline; and ampicillin, streptomycin, and tetracycline accounted for 22, 19, and 15%, respectively, of those identified. The data indicate a significant level of infectious drug resistance among the intestinal bacteria of the urban population.19695370461
5265170.9997Prevalence of antibiotic-resistant fecal bacteria in a river impacted by both an antibiotic production plant and urban treated discharges. In this study, the abundance and spatial dynamics of antibiotic-resistant fecal bacteria (Escherichia coli, total coliforms and Enterococcus spp.) were determined in water and sediment samples from a river impacted by both antibiotic production plant (APP) and urban wastewater treatment plant (WWTP) discharges. Agar dilution and disk diffusion methods were also used for antimicrobial susceptibility testing. Two antimicrobial agents, cephalexin (25 μg/ml) and amoxicillin (50 μg/ml), were evaluated using the agar dilution method for E. coli, total coliforms (TC) and Enterococcus spp., whereas the degree of sensitivity or resistance of E. coli isolates to penicillin (10 U), ampicillin (10 μg), doxycycline (30 μg), tetracycline (30 μg), erythromycin (15 μg), azithromycin (15 μg) and streptomycin (10 μg) was performed using the disk diffusion method. Real-time PCR assays were used to determine the prevalence of three antibiotic-resistance genes (ARGs). The agar dilution method showed that most E. coli isolates and TC were resistant to amoxicillin, especially after receiving the APP discharges. Antibiotic resistances to amoxicillin and cephalexin were higher after the APP discharge point than after the WWTP effluent. The disk diffusion method revealed that 100% of bacterial isolates were resistant to penicillin and erythromycin. Multidrug-resistant bacteria were detected and showed a higher proportion at the WWTP discharge point than those in the APP. Highly multidrug-resistant bacteria (resistance to more than 4 antibiotics) were also detected, reaching mean values of 41.6% in water samples and 50.1% in sediments. The relative abundance of the blaTEM, blaCTX-M and blaSHV genes was higher in samples from the treatment plants than in those collected upstream from the discharges, especially for water samples collected at the APP discharge point. These results clearly demonstrate that both the APP and the WWTP contribute to the emergence and spread of antibiotic resistance in the environment.201424836130
2686180.9997Antimicrobial resistance in Escherichia coli and Salmonella spp. isolates from fresh produce and the impact to food safety. Foodborne diseases associated with fresh produce consumption have escalated worldwide, causing microbial safety of produce of critical importance. Bacteria that have increasingly been detected in fresh produce are Escherichia coli and Salmonella spp., both of which have been shown to progressively display antimicrobial resistance. The study focused on the assessment of antimicrobial resistance of these enteric bacteria from different kinds of fresh produce from various open air markets and supermarkets in the Philippines. Using the disk diffusion assay on a total of 50 bacterial isolates obtained from 410 fresh produce surveyed, monoresistance to tetracycline was observed to be the most prevalent (38%), followed by multidrug resistance to tetracycline, chloramphenicol, ciprofloxacin, and nalidixic acid (4%), and lastly by dual resistance to tetracycline and chloramphenicol (2%). Using multiplex and simplex polymerase chain reaction (PCR) assays, tetA (75%) and tetB (9%) were found in tetracycline resistant isolates, whereas catI (67%) and catIII (33%) were detected in chloramphenicol resistant isolates. Sequence analysis of gyr and par genes from the ciprofloxacin and nalidixic acid resistant isolates revealed different mutations. Based on the results, fresh produce act as a reservoir of these antibiotic resistant bacteria which may pose health threat to consumers.201728679083
2691190.9997Antibiotic Resistant and Biofilm-Associated Escherichia coli Isolates from Diarrheic and Healthy Dogs. Bacteria isolated from companion animals are attracting concerns in a view of public health including antimicrobial resistance and biofilm development, both contributing to difficult-to-treat infections. The purpose of this study was to evaluate the minimum inhibitory concentrations (MIC) of 18 antibiotics in Escherichia coli isolated from two groups of dogs (healthy and diarrheic). Isolates were classified into phylogroups, examined for the presence of resistance genes and biofilm-formation capacity. In healthy dogs, phylogenetic analysis showed that 47.37% and 34.22% of E. coli isolates belonged to commensal groups (A; B1) in contrast to diarrheic dogs; 42.2% of isolates were identified as the B2 phylogroup, and these E. coli bacteria formed a stronger biofilm. The results of healthy dogs showed higher MIC levels for tetracycline (32 mg/L), ampicillin (64 mg/L), ciprofloxacin (8 mg/L) and trimethoprim-sulphonamide (8 mg/L) compared to clinical breakpoints. The most detected gene encoding plasmid-mediated resistance to quinolones in the healthy group was qnrB, and in dogs with diarrhea, qnrS. The resistance genes were more frequently detected in healthy dogs. The presence of the integron int1 and the transposon tn3 increases the possibility of transfer of many different cassette-associated antibiotic-resistance genes. These results suggest that dogs could be a potential reservoir of resistance genes.202134205399