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297900.9997Quinolone-resistant Escherichia coli in Poultry Farming. Increasing bacterial resistance to quinolone antibiotics is apparent in both humans and animals. For humans, a potential source of resistant bacteria may be animals or their products entering the human food chain, for example poultry. Between July 2013 and September 2014, samples were collected and analyzed in the Moravian regions of the Czech Republic to isolate the bacterium Escherichia coli. As a result, 212 E. coli isolates were obtained comprising 126 environmental isolates from poultry houses and 86 isolates from cloacal swabs from market-weight turkeys. Subsequently, the E. coli isolates were tested for susceptibility to selected antibiotics. Resistance of the poultry isolates to quinolones ranged from 53% to 73%. Additionally, the presence of plasmid-mediated resistance genes was studied. The genes were confirmed in 58% of the tested strains. The data on resistance of isolates from poultry were compared with results of resistance tests in human isolates obtained in the same regions. The high levels of resistance determined by both phenotyping and genotyping methods and reported in the present study confirm the fact that the use of fluoroquinolones in poultry should be closely monitored.201728662329
291010.9997Phenotypic and genotypic characterization of tetracycline and minocycline resistance in Clostridium perfringens. The aim of this study was to determine the incidence of tetracycline resistance and the prevalence of tetracycline-resistance genes in strains of Clostridium perfringens isolated from different sources between 1994 and 2005. Susceptibility to tetracycline and minocycline in strains from humans (35 isolates), chickens (15 isolates), food (21 isolates), soil (16 isolates) and veterinary sources (6 isolates) was determined, and tetracycline-resistance genes were detected. Resistance was most common in strains isolated from chickens, followed by those from soils, clinical samples and foods. The most highly resistant strains were found among clinical and food isolates. tetA(P) was the most common resistance gene, and along with tetB(P) was found in all resistant strains and some sensitive strains. One tetracycline-resistant food isolate had an intact tet(M) gene. However, PCR fragments of 0.4 or 0.8 kb with high degrees of identity to parts of the tet(M) sequences of other bacteria were found, mainly in clinical isolates, and often in isolates with tetB(P). No correlation between level of sensitivity to tetracycline or minocycline and the presence of tetA(P), tetB(P) or part of tet(M) was found. The presence of part of tet(M) in some strains of C. perfringens containing tetB(P) may have occurred by recent gene transfer.201020661548
555420.9997High prevalence of multidrug-tolerant bacteria and associated antimicrobial resistance genes isolated from ornamental fish and their carriage water. BACKGROUND: Antimicrobials are used to directly control bacterial infections in pet (ornamental) fish and are routinely added to the water these fish are shipped in to suppress the growth of potential pathogens during transport. METHODOLOGY/PRINCIPAL FINDINGS: To assess the potential effects of this sustained selection pressure, 127 Aeromonas spp. isolated from warm and cold water ornamental fish species were screened for tolerance to 34 antimicrobials. Representative isolates were also examined for the presence of 54 resistance genes by a combination of miniaturized microarray and conventional PCR. Forty-seven of 94 Aeromonas spp. isolates recovered from tropical ornamental fish and their carriage water were tolerant to > or =15 antibiotics, representing seven or more different classes of antimicrobial. The quinolone and fluoroquinolone resistance gene, qnrS2, was detected at high frequency (37% tested recent isolates were positive by PCR). Class 1 integrons, IncA/C broad host range plasmids and a range of other antibiotic resistance genes, including floR, bla(TEM-1), tet(A), tet(D), tet(E), qacE2, sul1, and a number of different dihydrofolate reductase and aminoglycoside transferase coding genes were also detected in carriage water samples and bacterial isolates. CONCLUSIONS: These data suggest that ornamental fish and their carriage water act as a reservoir for both multi-resistant bacteria and resistance genes.200920027306
292130.9997Diversity 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
593540.9997Antibiotic resistance genes in anaerobic bacteria isolated from primary dental root canal infections. Fourty-one bacterial strains isolated from infected dental root canals and identified by 16S rRNA gene sequence were screened for the presence of 14 genes encoding resistance to beta-lactams, tetracycline and macrolides. Thirteen isolates (32%) were positive for at least one of the target antibiotic resistance genes. These strains carrying at least one antibiotic resistance gene belonged to 11 of the 26 (42%) infected root canals sampled. Two of these positive cases had two strains carrying resistance genes. Six out of 7 Fusobacterium strains harbored at least one of the target resistance genes. One Dialister invisus strain was positive for 3 resistance genes, and 4 other strains carried two of the target genes. Of the 6 antibiotic resistance genes detected in root canal strains, the most prevalent were blaTEM (17% of the strains), tetW (10%), and ermC (10%). Some as-yet-uncharacterized Fusobacterium and Prevotella isolates were positive for blaTEM, cfxA and tetM. Findings demonstrated that an unexpectedly large proportion of dental root canal isolates, including as-yet-uncharacterized strains previously regarded as uncultivated phylotypes, can carry antibiotic resistance genes.201223108290
558250.9997Detection and prevalence of antimicrobial resistance genes in Campylobacter spp. isolated from chickens and humans. Campylobacter spp. are common pathogenic bacteria in both veterinary and human medicine. Infections caused by Campylobacter spp. are usually treated using antibiotics. However, the injudicious use of antibiotics has been proven to spearhead the emergence of antibiotic resistance. The purpose of this study was to detect the prevalence of antibiotic resistance genes in Campylobacter spp. isolated from chickens and human clinical cases in South Africa. One hundred and sixty one isolates of Campylobacter jejuni and Campylobacter coli were collected from chickens and human clinical cases and then screened for the presence of antimicrobial resistance genes. We observed a wide distribution of the tetO gene, which confers resistance to tetracycline. The gyrA genes that are responsible quinolone resistance were also detected. Finally, our study also detected the presence of the blaOXA-61, which is associated with ampicillin resistance. There was a higher (p < 0.05) prevalence of the studied antimicrobial resistance genes in chicken faeces compared with human clinical isolates. The tetO gene was the most prevalent gene detected, which was isolated at 64% and 68% from human and chicken isolates, respectively. The presence of gyrA genes was significantly (p < 0.05) associated with quinolone resistance. In conclusion, this study demonstrated the presence of gyrA (235 bp), gyrA (270 bp), blaOXA-61 and tetO antimicrobial resistance genes in C. jejuni and C. coli isolated from chickens and human clinical cases. This indicates that Campylobacter spp. have the potential of resistance to a number of antibiotic classes.201728582978
555560.9997New sequence types and multidrug resistance among pathogenic Escherichia coli isolates from coastal marine sediments. The spread of antibiotic-resistant microorganisms is widely recognized, but data about their sources, presence, and significance in marine environments are still limited. We examined 109 Escherichia coli strains from coastal marine sediments carrying virulence genes for antibiotic susceptibility, specific resistance genes, prevalence of class 1 and 2 integrons, and sequence type. Antibiotic resistance was found in 35% of strains, and multiple resistances were found in 14%; the resistances detected most frequently were against tetracycline (28%), ampicillin (16.5%), trimethoprim-sulfamethoxazole (13%), and streptomycin (7%). The highest prevalence of resistant strains was in phylogenetic group A, whereas phylogroup B2 exhibited a significantly lower frequency than all the other groups. Sixty percent of multiresistant strains harbored class 1 or 2 integrase genes, and about 50% carried resistance genes (particularly dfrA and aadA) linked to a class 1 integron. Multilocus sequence typing of 14 selected strains identified eight different types characteristic of extraintestinal pathogens and three new allelic combinations. Our data suggest that coastal marine sediment may be a suitable environment for the survival of pathogenic and antimicrobial-resistant E. coli strains capable of contributing to resistance spread via integrons among benthic bacteria, and they highlight a role for these strains in the emergence of new virulent genotypes.201222447595
204170.9997Carrier flies of multidrug-resistant Escherichia coli as potential dissemination agent in dairy farm environment. The life cycle of synanthropic flies and their behavior, allows them to serve as mechanical vectors of several pathogens. Given that flies can carry multidrug-resistant (MDR) bacteria, this study aimed to investigate the spread of genes of antimicrobial resistance in Escherichia coli isolated from flies collected in two dairy farms in Brazil. Besides antimicrobial resistance determinants, the presence of virulence genes related to bovine colibacillosis was also assessed. Of 94 flies collected, Musca domestica was the most frequently found in the two farms. We isolated 198 E. coli strains (farm A=135 and farm B=63), and >30% were MDR E. coli. We found an association between bla(TEM) and phenotypical resistance to ampicillin, or chloramphenicol, or tetracycline; and bla(CTX-M) and resistance to cefoperazone. A high frequency (86%) of phylogenetic group B1 among MDR strains and the lack of association between multidrug resistance and virulence factors suggest that antimicrobial resistance possibly is associated with the commensal bacteria. Clonal relatedness of MDR E. coli performed by Pulsed-Field Gel Electrophoresis showed wide genomic diversity. Different flies can carry clones, but with distinct antimicrobial resistance pattern. Sanger sequencing showed that the same class 1 integron arrangement is displayed by apparently unrelated strains, carried by different flies. Our conjugation results indicate class 1 integron transfer associated with tetracycline resistance. We report for the first time, in Brazil, that MDR E. coli is carried by flies in the milking environment. Therefore, flies can act as carriers for MDR strains and contribute to dissemination routes of antimicrobial resistance.201829758886
292280.9997Tetracycline-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
592190.9997Prevalence 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
2915100.9997Detection of class 1 integron-associated gene cassettes and tetracycline resistance genes in Escherichia coli isolated from ready to eat vegetables. BACKGROUND: Ready to eat (RTE) vegetables are easily accessible healthy foods that are commonly consumed globally, including in Indonesia. However, these RTE vegetables contain potential contamination from pathogens and multi-drug resistant bacteria. Therefore, in the present study, we examined the presence of tetracycline-resistant E. coli (TRE) isolates from RTE vegetables. METHODS: Susceptibility to antimicrobial agents was determined using the Kirby-Bauer disc diffusion method. Characterisation of antibiotic resistant genes was performed using PCR and sequencing of tetracycline resistant gene, integron and gene cassette from the TRE isolates. RESULTS: The isolates collected in this study were resistant not only to tetracycline, but also to streptomycin. Some isolates also displayed resistance to kanamycin (77.8%), chloramphenicol (11.1%), and ciprofloxacin (5.6%). All of the isolates contained integrons (intI1) and the tetA gene; tetB was not detected in our study. Further analysis showed that some isolates (38.8%) contained the dfrA7 gene cassette, which encodes dihydrofolate reductase, which is responsible for resistance to trimethoprim. Of all the isolates that presented integrons, 11 isolates (61.1%) did not carry gene cassettes. These empty integrons have the potential to convert themselves rapidly into multigraviton strains. CONCLUSIONS: TRE isolates contain the tetA gene and integron 1. Only 38.8% of the isolates that have been identified contain the dfrA7 gene cassette, which is responsible for trimethoprim antibiotic resistance. Further identification of genes conferring resistance to other antibiotics is necessary to better characterise antibiotic resistance.202032566218
5922110.9997Incidence 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
2050120.9997Identification of a novel fosfomycin resistance gene (fosA2) in Enterobacter cloacae from the Salmon River, Canada. AIMS: To investigate the occurrence of fosfomycin-resistant (fos(R) ) bacteria in aquatic environments. METHODS AND RESULTS: A fos(R) strain of Enterobacter cloacae was isolated from a water sample collected at a site (50°41'33·44″N, 119°19'49·50″W) near the mouth of the Salmon River at Salmon Arm, in south-central British Columbia, Canada. The strain was identified by PCR screening for plasmid-borne, fosA-family amplicons, followed by selective plating. Sequencing of the resistance gene cloned using PCR primers to conserved flanking DNA revealed a new allele (95% amino acid identity to fosA), and I-Ceu I PFGE showed that it was chromosomally located. In Escherichia coli, the cloned DNA conferred a greater resistance to fosfomycin than its fosA counterpart. CONCLUSIONS: Gene fosA2 conferred fosfomycin resistance in an environmental isolate of Ent. cloacae. SIGNIFICANCE AND IMPACT OF THE STUDY: The repurposing of older antibiotics should be considered in the light of existing reservoirs of resistance genes in the environment.201121392044
5553130.9997Detection of class 1 integrons in Salmonella Weltevreden and silent antibiotic resistance genes in some seafood-associated nontyphoidal isolates of Salmonella in south-west coast of India. AIMS: To study the antibiogram of 40 seafood isolates of Salmonella and use of PCR to detect the presence of integrons and genes coding for antibiotic resistance. METHODS AND RESULTS: In this study, 40 isolates of Salmonella were used for antibiogram analysis. The multidrug-resistant isolates were analyzed for the presence of integron using integron-specific primers. Twenty-five percentage of the isolates were multidrug resistant while 67·50% were resistant to at least two antibiotics. Antibiotic resistance genes catA1 and tetA were present in 57·52 and 60%, respectively. Although widespread presence of genes was observed, only 26·08% of the catA1-carrying isolates exhibited phenotypic resistance against the respective antibiotic. Integrons present in representative isolates of Salmonella Weltevreden and Salmonella Newport were sequenced. The former contained class 1 integron with a single gene dfrA7 in the integron cassette and an adjacent dihydropteroate synthetase gene along with the usual quaternary ammonium compound resistance gene, while the later contained class 1 integron with dhfrA1, OrfC, in the integron cassette and an adjacent dihydropteroate synthetase gene along with the usual quaternary ammonium compound resistance gene. CONCLUSIONS: This study demonstrates the presence of silent antibiotic resistance genes and class I integrons in seafood-associated Salmonella strains. The study also demonstrates the first report of class I integron in Salm. Weltevreden. Detection of catA1 genes in phenotypically sensitive bacteria suggests that these could be reservoirs in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The manuscript provides novel results describing the existence of a high rate of antibiotic resistance in the Salmonella populations prevailing in environmental sources as well as an absence of correspondence between the presence of antibiotic resistance genes, and the exhibition of a the corresponding phenotypic trait of resistance against the respective antibiotic compound was observed. In addition, the manuscript reports the presence of the class I integron in Salm. Weltevreden.201222443444
5583140.9997High prevalence of antimicrobial-resistant Escherichia coli from animals at slaughter: a food safety risk. BACKGROUND: There has been concern about the increase of antimicrobial resistant bacteria and protection of animal and public health, along with food safety. In the present study, we evaluate the incidence of antimicrobial resistance among 192 strains of Escherichia coli isolated from faecal samples of healthy food-producing animals at slaughter in Portugal. RESULTS: Ninety-seven % of the pig isolates, 74% from sheep and 55% from cattle were resistant to one or more antimicrobial agents, with the resistances to ampicillin, streptomycin, tetracycline and trimethoprim-sulfamethoxazole the most common phenotype detected. Genes encoding resistance to antimicrobial agents were detected in most of the resistant isolates. Ninety-three % of the resistant isolates were included in the A or B1 phylogenetic groups, and the virulence gene fimA (alone or in association with papC or aer genes) was detected in 137 of the resistant isolates. Five isolates from pigs belonging to phylogroup B2 and D were resistant to five different antimicrobial agents. CONCLUSION: Our data shows a high percentage of antibiotic resistance in E. coli isolates from food animals, and raises important questions in the potential impact of antibiotic use in animals and the possible transmission of resistant bacteria to humans through the food chain.201322836880
1606150.9997Salmonella spp. profiles isolated from seabird samples from the Brazilian coast. In view of growing concerns, in a One Health context, regarding the transport and dissemination of pathogenic microorganisms among seabirds and other vertebrate animals, including humans, the aim of this study was to identify Salmonella spp. in stranded and non-stranded resident and migratory wild seabirds from the Brazilian coast. Antimicrobial susceptibility and molecular profiles, quinolone resistance genes and antigenic characterization of the isolates were also carried out. Fresh faeces and cloacal swabs were obtained totaling 122 seabirds sampled throughout different Brazilian coast regions. At the laboratory, sample culturing, Salmonella spp. isolation and biochemical identification were performed, followed by antigenic profile identification by serum agglutination, susceptibility profile characterization by the agar disc diffusion technique, detection of quinolone resistance genes (qnrA, qnrB, qnrS) using the multiplex polymerase chain reaction technique (multiplex PCR) and, finally, isolates profiles identification by pulsed field gel electrophoresis (PFGE). Salmonella enterica subsp. enterica was identified in 7% of the studied birds, comprising three different serovars: Panama (63 %), Typhimurium (25 %) and Newport (13 %). The most important findings reported herein are the first description of Salmonella panama in seabirds and the totality of isolates being resistant (or intermediate) to at least one tested antimicrobial, with emphasis on quinolone resistance. The molecular results suggest that the observed resistance cannot be explained by the presence of plasmid-mediated quinolone resistance genes. The PFGE suggests that the Panama and Newport profiles detected herein are not yet widespread in Brazil, unlike Typhimurium, which is already well distributed throughout the country. Considering this finding, we suggest that seabirds are an important link in the epidemiological chain of this serovar. The monitoring of these bacteria in seabirds, as well as of their susceptibility profiles to antimicrobials, must be continuous, strengthening the role of these animals as environmental health indicators and sentinels.202134175569
5630160.9997Preliminary Results on the Prevalence of Salmonella spp. in Marine Animals Stranded in Sicilian Coasts: Antibiotic Susceptibility Profile and ARGs Detection in the Isolated Strains. The presence of Salmonella spp. in marine animals is a consequence of contamination from terrestrial sources (human activities and animals). Bacteria present in marine environments, including Salmonella spp., can be antibiotic resistant or harbor resistance genes. In this study, Salmonella spp. detection was performed on 176 marine animals stranded in the Sicilian coasts (south Italy). Antibiotic susceptibility, by disk diffusion method and MIC determination, and antibiotic resistance genes, by molecular methods (PCR) of the Salmonella spp. strains, were evaluated. We isolated Salmonella spp. in three animals, though no pathological signs were detected. Our results showed a low prevalence of Salmonella spp. (1.7%) and a low incidence of phenotypic resistance in three Salmonella spp. strains isolated. Indeed, of the three strains, only Salmonella subsp. enterica serovar Typhimurium from S. coeruleoalba and M. mobular showed phenotypic resistance: the first to ampicillin, tetracycline, and sulphamethoxazole, while the latter only to sulphamethoxazole. However, all strains harbored resistance genes (bla(TEM), bla(OXA), tet(A), tet(D), tet(E), sulI, and sulII). Although the low prevalence of Salmonella spp. found in this study does not represent a relevant health issue, our data contribute to the collection of information on the spread of ARGs, elements involved in antibiotic resistance, now considered a zoonosis in a One Health approach.202134451393
2696170.9997Carriage of antimicrobial resistant Escherichia coli in adult intestinal flora. Knowledge of antibiotic resistance in bacteria strains colonizing healthy people is important for several reasons, one of which is that; these organisms form one of the largest reservoirs of resistant genes. Frequency of resistance to eleven different antimicrobial agents was examined in faecal flora of adults with no history of recent antimicrobial treatment. Using the disc diffusion sensitivity test, 106 strains of Escherichia coli were examined, 68% of these were resistant to tetracycline, and 57% were resistant to ampicillin and cotrimoxazole respectively. There was no resistance to cefuroxime but resistance to ceftazidime was 13%. Fifty six out of the eighty eight (64%) isolates, which showed any resistance, were resistant to three or more antimicrobials. The most common resistant pattern was to three drugs tetracycline, ampicillin and cotrimoxazole. Six strains were susceptible to all antibiotics. One strain of Escherichia coli was resistant to eight antimicrobials. Thirty per cent of the Escherichia coli were resistant to gentamicin. This study reveals a high prevalence of resistant bacteria in faecal flora of healthy adults.200212081343
2797180.9997Widespread distribution of tetracycline resistance genes in a confined animal feeding facility. We sought to determine the distribution of resistance and the tetracycline resistance genes among bacteria isolated from a swine confined animal feeding facility where tetracycline-containing feed had been in use for over 20 years. Samples collected from feed, hogs, hog houses, waste lagoon, soil, surface water and well water were screened for the presence of (a) resistant Escherichia coli and enterococci and (b) tetracycline-resistant strains of all species. Genomic DNA was extracted from the latter strain collection and fragments from 16S rDNA and ten tetracycline resistance genes (tetA, tetB, tetC, tetE, tetH, tetL, tetM, tetS, tetT and rumB) were polymerase chain reaction-amplified and a partial nucleotide sequence was obtained. In this environment, 77% of E. coli and 68% of enterococci isolated were tetracycline resistant. Tetracycline resistance was found in 26 different bacterial genera and in 60 species. Single resistance gene alleles (as defined by nucleotide sequence) were present in multiple species. There was evidence of gene recombination and multiple different tetracycline resistance genes were present in single bacterial isolates. These data provide further evidence for the widespread distribution of resistance genes in microbial populations in settings in which there is ongoing subtherapeutic antimicrobial use.200717287111
5926190.9997Prevalence and Characterization of Gentamicin Resistance Genes in Escherichia coli Isolates from Beef Cattle Feces in Japan. Gentamicin is an important antibiotic for the treatment of opportunistic infections in the clinical field. Gentamicin-resistant bacteria have been detected in livestock animals and can be transmitted to humans through the food supply or direct contact. We have previously revealed that gentamicin-resistant Escherichia coli are distributed at a comparatively high rate from beef cattle in Japan, but few studies have focused on the molecular epidemiology of gentamicin-resistant bacteria. To understand these bacteria, this study examined the prevalence of various gentamicin resistance genes in gentamicin-resistant E. coli isolates from beef cattle feces. Of the 239 gentamicin-resistant E. coli isolates, the presence of the aacC2, aadB, or aac(3)-VIa genes was confirmed in 147, 84, and 8 isolates, respectively. All aac(3)-VIa-harboring isolates had an MIC value of 64 μg/mL for gentamicin and exhibited resistance to 11 antibiotic agents. An analysis of the representative aac(3)-VIa-harboring E. coli strain GC1-3-GR-4 revealed that the aac(3)-VIa gene was present on the IncA/C plasmid together with the aadA and bla(CMY) genes. Furthermore, the upstream region of the aac(3)-VIa gene contained the aadA gene and the class 1 integron-integrase gene (intI1). The aac(3)-VIa gene was detected for the first time in Japan and is expected to be able to transfer between bacteria via the IncA/C plasmid and integron. These results reveal the expansion of the distribution or diversity of gentamicin resistance genes in Japan.202235704076