Browsing by Author "Correia, Ivone"
Now showing 1 - 9 of 9
Results Per Page
Sort Options
- Antimicrobial susceptibility and oxymino-β-lactam resistance mechanisms in Salmonella enterica and Escherichia coli isolates from different animal sourcesPublication . Clemente, Lurdes; Manageiro, Vera; Jones-Dias, Daniela; Correia, Ivone; Themudo, Patricia; Albuquerque, Teresa; Geraldes, Margarida; Matos, Filipa; Almendra, Cláudia; Ferreira, Eugénia; Caniça, ManuelaThe impact of extended-spectrum β-lactamases (ESBLs) and plasmid-mediated AmpC β-lactamases (PMAβs) of animal origin has been a public health concern. In this study, 562 Salmonella enterica and 598 Escherichia coli isolates recovered from different animal species and food products were tested for antimicrobial resistance. Detection of ESBL-, PMAβ-, plasmid-mediated quinolone resistance (PMQR)-encoding genes and integrons was performed in isolates showing non-wild-type phenotypes. Susceptibility profiles of Salmonella spp. isolates differed according to serotype and origin of the isolates. The occurrence of cefotaxime non-wild-type isolates was higher in pets than in other groups. In nine Salmonella isolates, blaCTX-M (n = 4), blaSHV-12 (n = 1), blaTEM-1 (n = 2) and blaCMY-2 (n = 2) were identified. No PMQR-encoding genes were found. In 47 E. coli isolates, blaCTX-M (n = 15), blaSHV-12 (n = 2), blaCMY-2 (n = 6), blaTEM-type (n = 28) and PMQR-encoding genes qnrB (n = 2), qnrS (n = 1) and aac(6')-Ib-cr (n = 6) were detected. To the best of our knowledge, this study is the first to describe the presence of blaCMY-2 (n = 2) and blaSHV-12 (n = 1) genes among S. enterica from broilers in Portugal. This study highlights the fact that animals may act as important reservoirs of isolates carrying ESBL-, PMAβ- and PMQR-encoding genes that might be transferred to humans through direct contact or via the food chain.
- Antimicrobial susceptibility of Escherichia coli strains isolated from food-producing, companion and wild animals, in Portugal – Characterization of isolates with reduced susceptibility to third generation cephalosporins and cephamycinsPublication . Clemente, Lurdes; Manageiro, Vera; Jones-Dias, Daniela; Ferreira, Eugénia; Correia, Ivone; Albuquerque, Teresa; Geraldes, Margarida; Matos, Filipa; Almendra, Claúdia; Themudo, Patrícia; Caniça, ManuelaEscherichia coli is a common inhabitant of the gastrointestinal tract of humans and animals, but is also a causative agent of diarrhea and extraintestinal infections. There is ongoing concern about the risks posed to human health by antimicrobial resistant bacteria isolated from animals. This study was conducted on 602 Escherichia coli isolates recovered from food-producing (n=217), companion (n=114) and wild animals (n=271), over the period of 2009-2013, to determine their antimicrobial susceptibility to a panel of ten antimicrobials (ampicillin, cefotaxime, ciprofloxacin, nalidixic acid, trimethoprim, sulfamethoxazole, chloramphenicol, streptomycine, gentamicine and tetracycline), through the determination of Minimum Inhibitory Concentration (MIC), using the agar dilution technique. Susceptibility to cefoxitin was determined through disk diffusion method. Molecular characterization of isolates showing a non-wild type MIC to cefotaxime was performed, to determine extended spectrum β-lactamases (ESBL), plasmid mediated AmpC (PMAβ), plasmid mediated quinolone (PMQR) resistance determinants and mobile genetic elements involved in the dissemination of resistance genes. Overall, isolates recovered from food producing animals showed higher frequencies of resistance towards all antimicrobials tested and multidrug resistance (MDR) (53%), followed by companion (43%) and wild animals (30%). Fifty isolates presented a non-wild phenotype to cefotaxime and resistance or intermediate susceptibility to cefoxitin, being 14 (12.3%) from pets, 19 (8.8%) from food producing animals and 17 (6.3%) from wild animals. Polymerase chain reaction and sequencing of the amplicons confirmed the presence of blaCTX-M-type (n=14), blaSHV-type (n=2), blaTEM-type (n=31), blaOXA-type (n=6) and plasmid-mediated AmpC β-lactamases (PMAβ) genes (n=8). Plasmid-mediated quinolone resistance-encoding genes were detected in ten isolates: aac(6')-Ib-cr (n=6) qnrB17 (n=1), qnrS1(n=1) and qnrB19 (n=2). Twenty five isolates carried class 1 integrons and two carried class 2 integrons. This study agrees that animals may act as important reservoirs of E. coli isolates carrying ESBL and PMAβ-encoding genes, which might be transmissible to humans through direct contact or the food chain and, a potential source for human pathogens to acquire these resistance genes.
- Antimicrobial susceptibility of Salmonella enterica isolated from food-producing animals, animal feed and food products of animal origin, in Portugal - Genetic analysis of isolates with reduced susceptibility/resistance to third generation cephalosporins and cephamycinsPublication . Clemente, Lurdes; Manageiro, Vera; Jones-Dias, Daniela; Ferreira, Eugénia; Correia, Ivone; Themudo, Patrícia; Albuquerque, Teresa; Caniça, ManuelaSalmonella is a widely distributed foodborne pathogen and one of the most common causes of bacterial foodborne illnesses in humans. An epidemiologic study was conducted on 1600 Salmonella spp isolates recovered from poultry, swine, other animal species, animal feed and food products of animal origin, over the period of 2009-2013, to determine their serotype and antimicrobial susceptibility to a panel of ten antimicrobials (ampicillin, cefotaxime, ciprofloxacin, nalidixic acid, trimethoprim, sulfamethoxazole, chloramphenicol, streptomycine, gentamicine and tetracycline), through the determination of Minimum Inhibitory Concentration (MIC), using the agar dilution technique. Molecular characterization of isolates showing a non-wild type MIC to cefotaxime was performed, to determine extended spectrum β-lactamases (ESBL), plasmid mediated AmpC (PMAβ), plasmid mediated quinolone (PMQR) resistance determinants and mobile genetic elements involved in the dissemination of resistance genes. In live poultry (breeders, broilers, layers) of the 843 isolates recovered, 27.9% comprised S. Enteritidis, 23,5% Salmonella Havana and 14.1% Salmonella Mbandaka; in turkeys, Salmonella Derby was the most common serovar isolated (44%), followed by Salmonella I 4,[5],12:i:- (16%). In swine, of 101 isolates 21.8% comprised Salmonella Rissen and Salmonella Typhimurium, 10.9% Salmonella Derby and Salmonella London. In other animal species, Salmonella Typhimurium was the prevalent serovar with 65.6% of the isolates, followed by Salmonella I 4,[5],12:i:- (9.8%). Overall, S. 4,[5],12:i:- was the most common serotype recovered from food products (25.8%), followed by S. Typhimurium (19.2%) and Salmonella Rissen (18.4%). S. Enteritidis was the most frequent serotype in poultry products (36.3%). Susceptibility profiles differed according with the serotype and the origin of the isolates. A higher frequency of multidrug resistant isolates was recovered from food of swine and bovine origin, with 62.6% and 59.4%, respectively. Polymerase chain reaction and sequencing of the amplicons confirmed the presence of blaCTX-M-type (n=8), blaSHV-type (n=2), blaTEM-type (n=2) and plasmid-mediated AmpC β-lactamases (PMAβ) genes (n=2). No plasmid mediated quinolone resistance-encoding genes were detected. Six isolates( three S. I 4,[5],12:i:-, two S. Havana and one S. Enteritidis) carried class 1 integrons and one S. I 4,[5],12:i:- isolate harboured a class 2 integron. In conclusion, the growing concern of the emergence of bacterial strains bearing ESBL in food-producing animals highlights the importance of continuous monitoring.
- Antimicrobial susceptibility of Salmonella enterica isolates from healthy breeder and broiler flocks in PortugalPublication . Clemente, Lurdes; Correia, Ivone; Themudo, Patrícia; Neto, Isabel; Caniça, Manuela; Bernardo, FernandoThree hundred and thirty-three isolates representing 40 different serotypes of Salmonella enterica, recovered from environmental and faecal samples of breeder and broiler flocks from 2009 to 2011, were studied. Antimicrobial susceptibility was determined by measuring the minimal inhibitory concentration of 11 antimicrobials using the agar dilution method. Salmonella Havana, S. Enteritidis and S. Mbandaka were the most common serotypes isolated from broiler flocks, while S. Enteritidis was the common isolate from breeder flocks. The frequency of non-wild-type Salmonella isolates (those with decreased susceptibility to the different antimicrobials) varied according to serotype. S. Mbandaka in broilers and S. Enteritidis in both breeders and broilers showed higher frequencies of reduced susceptibility to quinolones, but clinical resistance towards ciprofloxacin was not observed. Reduced susceptibility to sulfamethoxazole, tetracycline, ampicillin and streptomycin were common in Salmonella Typhimurium isolates. Two isolates of S. Havana from broilers were resistant to cefotaxime and phenotypically categorised as extended-spectrum β-lactamase producers. The results presented in this study provide useful data on the antimicrobial susceptibility of different Salmonella serotypes and highlight the high diversity of multi-drug resistance patterns present.
- 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.
- New insights into resistance to colistin and third-generation cephalosporins of Escherichia coli in poultry, Portugal: Novel blaCTX-M-166 and blaESAC genesPublication . Manageiro, Vera; Clemente, Lurdes; Graça, Rafael; Correia, Ivone; Albuquerque, Teresa; Ferreira, Eugénia; Caniça, ManuelaThe increasing incidence of intestinal colonization with extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae and Gram negative organisms that has been observed in food animals such as poultry, cattle and pigs, are suggestive that animals, food and environment are potential sources of ESBL-producing bacteria. Hence, the aim of this study was to characterized commensal E. coli obtained from healthy broiler and turkey flocks at slaughter for the presence of penicillinases-, ESBL-, extended-spectrum AmpC (ESAC)-, plasmid-mediated quinolone resistance- and MCR-encoding genes. Study of clonal relatedness showed genetic diversity among CTX-M-type, SHV-12 and TEM-52 producing isolates with human isolates of the same type, was also assessed. We detected that eleven (5.4%, 11/202) and forty-five (2.2%, 45/185) E. coli isolates from broilers and turkeys, respectively, carried blaESBLor blaESACgenes and two isolates from turkeys carried mcr-1 gene. A new variant blaCTX-M-166was reported in a multidrug resistant isolate from a broiler flock. Overall, we detected a diversity of resistance mechanisms among E. coli from food-producing animals, all of them with high importance at a public health level.
- 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 extended-spectrum β-lactamases among isolates of Salmonella enterica subsp. enterica from food-producing animals and food products, in PortugalPublication . Clemente, Lurdes; Manageiro, Vera; Ferreira, Eugénia; Jones-Dias, Daniela; Correia, Ivone; Themudo, Patrícia; Albuquerque, Teresa; Caniça, ManuelaA total of 1120 Salmonella spp. isolates, recovered from poultry, swine and food products of animal origin (bovine, swine and poultry) over the period of 2009-2011, were investigated in order to determine their serotype, susceptibility to a panel of eleven antimicrobials (A, ampicillin; Ct, cefotaxime; Cp, ciprofloxacin; Tm, trimethoprim; Su, sulfamethoxazole; C, chloramphenicol; S, streptomycin; G, gentamicin; T, tetracycline; NA, nalidixic acid; Fl, florfenicol), and the presence of resistance determinants of extended-spectrum cephalosporins. Overall, Salmonella Enteritidis was the most common serotype in all three animal species. In 618 isolates of poultry, 32.8% comprised S. Enteritidis, 18.3% Salmonella Havana and 16.5% Salmonella Mbandaka; in 101 isolates of pigs, 21.8% comprised Salmonella Rissen and Salmonella Typhimurium, 10.9% Salmonella Derby and Salmonella London. Salmonella I 4,[5],12:i:- was the most common serotype recovered from pork and beef food products comprising 32.6% and 30% of isolates respectively, followed by S. Rissen (26% and 24%) and S. Typhimurium (18.2% and 19%), respectively. In poultry products, S. Enteritidis was the most frequent serotype (62.7%), followed by S. Mbandaka (10.2%) and S. Derby (8.5%). Susceptibility profiles differed according to the origin of the isolates. Five multidrug resistant isolates (0.45%) were further characterized as extended-spectrum β-lactamase (ESBL) producers. Polymerase chain reaction and sequencing of the amplicons confirmed the presence of bla(CTX-M-1) (n = 2), bla(CTX-M-14) (n = 1), bla(CTX-M-15) (n = 1) and bla(CTX-M-32) (n = 1); bla(SHV-12) and bla(TEM-1) genes were also detected in two isolates of S. I 4,[5],12:i:-. Four isolates, two S. Havana and two S. I 4,[5],12:i:-, carried class 1 integrons and in three, two S. I 4,[5],12:i:- and one S. Havana, ISEcp1 was identified associated to bla(CTX-M-1), bla(CTX-M-32) and bla(CTX-M-14) genes. Additionally, in one S. I 4,[5],12:i:- isolate, orf477 was identified linked to bla(CTX-M-32). No plasmid mediated quinolone resistance-encoding genes were detected. Here, we report for the first time the presence of bla(CTX-M) genes in Salmonella enterica subsp enterica isolates recovered from poultry and food products of swine origin, in Portugal.
- Revealing mcr-1-positive ESBL-producing Escherichia coli strains among Enterobacteriaceae from food-producing animals (bovine, swine and poultry) and meat (bovine and swine), Portugal, 2010-2015Publication . Clemente, Lurdes; Manageiro, Vera; Correia, Ivone; Amaro, Ana; Albuquerque, Teresa; Themudo, Patrícia; Ferreira, Eugénia; Caniça, ManuelaWe screened 1840 Enterobacteriaceae isolates from food-producing animals, meat, meat products and animal feed, for the detection of plasmid-mediated colistin resistance, during 2010-2015. The mcr-1 gene was detected in 8.0% (97/1206) Escherichia coli and in 0.47% (3/634) Salmonella enterica isolates, with a high number of mcr-1 positive E. coli isolates (45.7%) being extended-spectrum β-lactamase or plasmid-mediated AmpC β-lactamase co-producers. No mcr-2 gene was detected. Our findings highlight the spread of mcr-1 genes within a wide-ranging sample of food-producing animals and meat, in Portugal.