DDI - Posters/abstracts em congressos internacionais
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Browsing DDI - Posters/abstracts em congressos internacionais by Author "Albuquerque, Teresa"
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- Diversity of β-lactamase-encoding genes in Escherichia coli strains isolated from food-producing, companion and zoo animals in PortugalPublication . Clemente, Lurdes; Correia, Ivone; Albuquerque, Teresa; Geraldes, Margarida; Matos, Filipa; Themudo, Patrícia; Manageiro, Vera; Jones-Dias, Daniela; Ferreira, Eugénia; Caniça, ManuelaA rapid development of plasmid-mediated resistance to extended-spectrum cephalosporins has been observed in Enterobacteriaceae worldwide, predominantly due to the dissemination of extended-spectrum beta-lactamases (ESBL) and plasmid-mediated AmpC beta-lactamases (PMAB). The aim of the present study was to evaluate the extension of ESBL- and PMAB-producing E. coli strains isolated from different animal origins in Portugal. For surveillance purposes, 376 E. coli isolates identified at National Laboratory of Veterinary Research (2009-2011) were submitted to antimicrobial susceptibility testing: 123, 51 and 202 were isolated from food-producing, companion and zoo animals, respectively. Minimum Inhibitory Concentrations (MIC) of 11 antimicrobials for all isolates was determined through agar dilution method. Susceptibility towards cefoxitina was determined through disk diffusion method. Breakpoints were interpreted accordingly to EUCAST epidemiological cut-off values. ‘Non-wild type’ (NWT) isolates for cefotaxime (MIC>0.25mg/L) and/or cefoxitina (<19mm) were screened for the presence of ESBL (blaTEM, blaOXA, blaSHV, blaCTX) and PMAB encoding genes, using PCR method. Sequencing was applied to fully identify beta-lactamases. Seventeen isolates (4.5%) were ‘NWT’ strains for cefotaxime, being 5 (29.4%) from companion animals, 4 (23.5%) from food-producing animals and 8 (47.1%) from zoo animals. We identified blaCTX-M-14 (n=1) in a dog and blaCTX-M-15-type genes (n=9) in 6 zoo animals and 3 in food-producing animals. We also identified blaCMY-type genes (n=3) in ‘NWT’ isolates for cefoxitin, one from each animal category. Other beta-lactamase encoding genes were identified: blaOXA in 5 strains (29.4%) isolated from dolphins, blaTEM in 7 strains (41.2%) isolated from 3 companion animals, 2 food-producing and 2 zoo animals, and blaSHV identified in one isolate (5.9%) from a zoo animal; 13 beta-lactamase-producing isolates (76.5%) were multidrug resistant. Among ‘NWT’ E. coli isolates for cefotaxime, we identified an important diversity of ESBL encoding genes, belonging to different families, being blaCTX-M-15-type gene the predominant. The spread of ESBL-producing bacteria among species from different origins, such as food-producing, companion and zoo animals, is a concern at public health level. Thus, it should be a priority to monitor and identify the reservoirs of antimicrobial resistance, contributing to a single health for all.
- Epidemiology and zoonotic potential of Escherichia coli CTX-M-15-producing isolates in PortugalPublication . Caniça, Manuela; Clemente, Lurdes; Jones-Dias, Daniela; Manageiro, Vera; Themudo, Patrícia; Albuquerque, Teresa; Francisco, Ana Patrícia; Louro, Deolinda; Ferreira, EugéniaBackground: The recent spread of plasmid-located CTX-M-ESBL-encoding genes is a serious threat to the clinical efficacy of expanded-spectrum cephalosporins. This study proposes to identify the epidemiology of plasmid-mediated CTX-M-encoding genes between an Escherichia coli strain isolated from a dolphin and several E. coli strains of human origin and, explain the responsible mechanism and reservoirs of current spread of CTX-M-type enzymes. Molecular typing of these strains establishes the linkage of dissemination between human and animal isolates. Methods: Sixty two ESBL-positive E. coli strains isolated from different clinical specimens in seven hospitals (2004 to 2009), from four different geographic regions, were screened for the presence of CTX-M encoding genes. An E. coli isolated from a respiratory exsudate in a dolphin in 2009, at the National Laboratory of Veterinary Research (LNIV) and characterized as CTX-M-producer, was also included in this study. Antimicrobial susceptibility was performed by broth-microdilution method. PCR and sequencing were used to screen and identify bla genes. Genetic relatedness among all isolates was examined by PFGE using XbaI enzyme. MLST was performed among the clinical epidemic human isolates clustering together and the isolate from the dolphin, according to the MLST database. Results: Forty eight human clinical isolates (77%) were CTX-M producers. Susceptibility towards beta-lactams confirmed all isolates as ESBL producers and also suggesting CTX-M enzymes expression. We detected blaCTX-M-15 (n=34), blaCTX-M-1 (n=4), blaCTX-M-3 (n=3), blaCTX-M-32 (n=3), blaCTX-M-14 (n=4), blaTEM-1 (n=39), and blaSHV-12 (n=8) genes; the dolphin isolate presented the blaCTX-M-15 gene. Genetic relatedness analysis by PFGE revealed one major cluster corresponding to a single epidemic clone A, which included 22 (35%) of all human isolates and the dolphin isolate, which clustered together with this clone A and, exhibited the same combination of MLST alleles across the seven sequenced loci, corresponding to the ST131. Twenty-one clinical isolates corresponding to the CTX-M-15-positive clone A were multidrug-resistant (95%), as well as the dolphin isolate. Conclusions: This study illustrated that genetic relatedness between human and animal E. coli CTX-M-15-producing clone strengthens its zoonotic potential. It may also explain the current spread of CTX-M-type enzymes worldwide among species from different origins and highlights the importance to identify antimicrobial resistance reservoirs, contributing to a single health for all.
- Occurrence of extended-spectrum beta-lactamases in Salmonella enterica strains isolated from broilers and food of animal origin in PortugalPublication . Clemente, Lurdes; Correia, Ivone; Themudo, Patrícia; Albuquerque, Teresa; Manageiro, Vera; Jones-Dias, Daniela; Ferreira, Eugénia; Caniça, ManuelaSalmonella enterica is a zoonotic bacteria transmitted through the food chain and isolates harbouring extended-spectrum beta-lactamases (ESBLs) have emerged worldwide during the last decade, with the CTX-M group being particularly important. The aim of the present study was to determine the antimicrobial susceptibility of S. enterica strains isolated from broilers and food of animal origin and to characterize ESBLs producers. On the scope of the national antimicrobial resistance surveillance programme on Salmonella, a total of 283 strains isolated from broilers (n=100) and food of animal origin (n=183), were received at the National Laboratory of Veterinary Research in 2011. The minimum inhibitory concentration (MIC) of 11 antimicrobials (nalidixic acid, ciprofloxacin, ampicillin, cefotaxime, chloramphenicol, florfenicol, streptomycin, gentamicin, tetracycline, sulphamethoxazole and trimethoprim) for all isolates was determined by agar dilution method. Susceptibility towards cefoxitin was determined through disk diffusion method. Breakpoints were interpreted accordingly to EUCAST epidemiological cut-off values. ‘Non-wild type’ (‘NWT’) isolates for cefotaxime (MIC>0.5mg/L) and cefoxitin (<19mm) were screened for the presence of ESBL- (blaTEM, blaOXA, blaSHV, blaCTX) and PMA_-encoding genes, using PCR method. Sequencing was applied to fully identify beta-lactamases. Among broilers, we identified 62% of ‘NWT’ isolates for ciprofloxacin, 57% for nalidixic acid and 28% for sulphamethoxazole, whereas in isolates from food of animal origin, 71%, 63% and 56% were ‘NWT’ isolates for tetracycline, sulphamethoxazole and ampicillin, respectively. Among all, 5/283 (1.8%) strains presented ‘NWT’ MICs for cefotaxime and were multidrug resistant: 2 Salmonella Havana isolated from broilers and 3 Salmonella S. 4,[5],12:i:- isolated from food of animal origin (swine); these isolates had one blaCTX-M-type gene, and 2 from food of animal origin presented 1 blaTEM-type gene and 1 blaSHV-type gene, respectively; they were ‘wild type’ for cefoxitin and no PMAB-encoding gene was detected. To our knowledge, this is the first time in Portugal that ESBL-encoding genes, particularly from blaCTXM family, were detected in isolates of Salmonella Havana, a very common serotype isolated from our broiler population. It should also be emphasised that third generation cephalosporins are not allowed in the national poultry production, contrary to the large animal production, which may explain the detection of ESBL-encoding genes in our strains from swine origin. Horizontal gene transfer may be responsible for the coresistance of strains to non-beta-lactam antibiotics. This study shows that national animal health monitoring systems play an important role and should be improved in an international level.
- Occurrence of plasmid-mediated quinolone resistance among bacteria isolated in animals in PortugalPublication . Ferreira, Eugénia; Clemente, Lurdes; Jones-Dias, Daniela; Francisco, Ana Patrícia; Manageiro, Vera; Themudo, Patrícia; Albuquerque, Teresa; Amado, Alice; Caniça, ManuelaBackground: Plasmid-mediated quinolone resistance (PMQR) is increasingly identified worldwide in Enterobacteriaceae. The aim of this study was to evaluate the extension of PMQR in isolates from animals in Portugal. Methods: We screened 186 Enterobacteriaceae isolates for the presence of PMQR determinants, identified at National Laboratory of Veterinary Research (2008-2009). A total of 92 Salmonella isolates were isolated from broilers, layers and pigs, and 94 Escherichia coli were from farm animals, birds and mammals. Susceptibility testing of all isolates was performed by disk diffusion method, and MICs against nalidixic acid, ciprofloxacin, gatifloxacin, levofloxacin, ofloxacin, enrofloxacin, morbifloxacin and norfloxacin were determined by E-test for PMQR-positive isolates. PCR and nucleotide sequencing, by using specific primers, were used to screen for the presence of PMQR-encoding genes. The genetic context of PMQR genes was evaluated by using different molecular methods. Results: We identified 5 qnrC-positive isolates: 2 Salmonella enteritidis collected in 2 layer chicken and in 3 E. coli from 2 broilers and one pig; 3 qnrS1 genes were detected in E. coli isolates from a broiler (co-expressing a qnr-C gene), a dog and a turtle-dove. The aac-(6’)-Ib-cr gene was detected in an E. coli isolated from a mammalian. Seven PMQR-positive isolates showed diminished susceptibility to at least one quinolone, and one was detected in the range of susceptibility against the seven (fluoro)quinolones tested. Three E. coli and one S. enteritidis were PMQR- and TEM-1 and/or CTX-M-15-producing isolates. An E. coli with qnrC, qnrS1, and blaTEM-1 genes and an E. coli with qnrC gene were positive for genes coding to class 1 integrons. Conclusions: This survey showed that PMQR determinants are present in animals from different environments in Portugal, including food-producing animals, with a high frequency (3%) of QnrC-producing isolates. Susceptibility results demonstrate the difficulty to predict the PMQR mechanisms by phenotypic methods. Overall, the study suggests that PMQR genes are undergoing a dissemination process, which needs surveillance.
- Zoonotic potential of multidrug resistant Escherichia coli clonal groups in PortugalPublication . Jones-Dias, Daniela; Clemente, Lurdes; Manageiro, Vera; Themudo, Patrícia; Albuquerque, Teresa; Francisco, Ana Patrícia; Louro, Deolinda; Ferreira, Eugénia; Caniça, ManuelaMultidrug-resistant (MDR) plasmids together with clonal dissemination of resistant isolates are possibly the most successful combination of factors contributing to the spread of antibiotic resistance genes. Our aim was to identify clones associated with multidrug-resistance among human and dolphin isolates, in order to evaluate zoonotic potential and risk. Sixty two MDR human Escherichia coli isolates were randomly selected from NIH collection, being previously isolated from different clinical specimens in seven geographically apart Portuguese hospitals from 2004 to 2009. Two E. coli isolated from dolphin's respiratory exudates in 2009 and 2010, at the National Laboratory of Veterinary Research, were also included in this study for their zoonotic portencial analysis. Antimicrobial susceptibility was performed by broth-microdilution method (EUCAST). PCR and sequencing were used to screen and identify β-lactamase and Aac(6')-Ib-cr encoding genes, while PCR-based replicon typing was used to characterize plasmids from MDR isolates. Genetic relatedness of human and dolphin isolates was examined both by PFGE and MLST. Mobile genetic elements were also investigated through PCR mapping assays. Regarding the human isolates, 48 (77%) were CTX-M producers. We detected blaCTX-M-1 (n=4), blaCTX-M-3 (n=3), blaCTX-M-14, blaCTX-M-15 (n=34), blaCTX-M-32 (n=3), (n=4), blaTEM-1 (n=39), and blaSHV-12 (n=8) genes as well as aac(6')-Ib-cr (n=26). Concerning the isolates recovered from the dolphins, one of them produced TEM-1, OXA-30, CTX-M-15 and Aac(6')-Ib-cr and the other TEM-1, Aac(6')-Ib and Aac(6')-Ib-cr. Replicon-typing revealed a severe predominance of IncF plasmids in both animal and human isolates; IS26 and ISEcp1 were also detected in both groups, being associated with blaCTX-M-15 and Aac(6')-Ib-cr plus OXA-30, respectively, in one of the dolphin isolates. Genetic relatedness analysis by PFGE revealed one major cluster corresponding to a single epidemic clone A, which included 22 (35%) of all human isolates and both dolphin isolates. They exhibited the same combination of MLST alleles, corresponding to ST131. This study illustrated the dominance of common antibiotic resistance genes, plasmids and clonal groups, specifically blaCTX-M-15, aac(6')-Ib-cr, IncF plasmids and ST131 in both human and animal isolates, reflecting their linkage and enhancing their zoonotic potencial. Studies should be performed to further deepen their role as hotspots of resistance.
- Zoonotic potential of multidrug resistant Escherichia coli clonal groups in PortugalPublication . Jones-Dias, Daniela; Clemente, Lurdes; Manageiro, Vera; Themudo, Patrícia; Albuquerque, Teresa; Francisco, Ana Patrícia; Louro, Deolinda; Ferreira, Eugénia; Caniça, ManuelaObjectives: Multidrug-resistant (MDR) plasmids together with clonal dissemination of resistant isolates are possibly the most successful combination of factors contributing to the spread of antibiotic resistance genes. Our aim was to identify clones associated with multidrug-resistance among human and dolphin isolates, in order to evaluate zoonotic potential and risk. Methods: Sixty two MDR human Escherichia coli isolates were randomly selected from NIH collection, being previously isolated from different clinical specimens in seven geographically apart Portuguese hospitals from 2004 to 2009. Two E. coli isolated from dolphin’s respiratory exudates in 2009 and 2010, at the National Laboratory of Veterinary Research, were also included in this study for their zoonotic potencial analysis. Antimicrobial susceptibility was performed by broth-microdilution method (EUCAST). PCR and sequencing were used to screen and identify beta-lactamase and Aac(6’)-Ib-cr encoding genes, while PCR-based replicon typing was used to characterize plasmids from MDR isolates. Genetic relatedness of human and dolphin isolates was examined both by PFGE and MLST. Mobile genetic elements were also investigated through PCR mapping assays. Results: Regarding the human isolates, 48 (77%) were CTX-M producers. We detected blaCTX-M-1 (n = 4), blaCTX-M-3 (n = 3), blaCTX-M-14, blaCTX-M-15 (n = 34), blaCTX-M-32 (n = 3), (n = 4), blaTEM-1 (n = 39), and blaSHV-12 (n = 8) genes as well as aac(6’)-Ib-cr (n = 26). Concerning the isolates recovered from the dolphins, one of them produced TEM-1, OXA-30, CTX-M-15 and Aac(6’)-Ib-cr and the other TEM-1, Aac(6¢)-Ib and Aac(6’)-Ib-cr. Replicon-typing revealed a severe predominance of IncF plasmids in both animal and human isolates; IS26 and ISEcp1 were also detected in both groups, being associated with blaCTX-M-15 and Aac(6’)-Ib-cr plus OXA-30, respectively, in one of the dolphin isolates. Genetic relatedness analysis by PFGE revealed one major cluster corresponding to a single epidemic clone A, which included 22 (35%) of all human isolates and both dolphin isolates. They exhibited the same combination of MLST alleles, corresponding to ST131. Conclusion: This study illustrated the dominance of common antibiotic resistance genes, plasmids and clonal groups, specifically blaCTX-M-15, aac(6’)-Ib-cr, IncF plasmids and ST131 in both human and animal isolates, reflecting their linkage and enhancing their zoonotic potencial. Studies should be performed to further deepen their role as hotspots of resistance.