Browsing by Author "Pista, Angela"
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- Are Food Producing animals a source of multidrug-resistant E. coli and Salmonella spp.?Publication . Pista, Angela; Silveira, Leonor; Gonçalves, Carlota; Costa, Sara; Carolino, Catarina; Santo, Bianca; Rosa, Rute; Penteado, Margarida; Alves, Margarida; Belas, Adriana; Santos, Isabel; Lima, Ana; Mota, Joana; Pedroso, Laurentina; Gomes, João Paulo; Nunes, Alexandra; Ramos, SóniaObjectives: To assess the role of food-producing animals as potential transmission vehicles of Salmonella spp. and E. coli to Humans, in order to understand the epidemiology and population structure of these zoonotic agents in Portugal.
- Aumento de casos de Salmonella Typhimurium em 2024: caracterização fenotípica e genotípica dos isoladosPublication . Tomás, Alexandra; Henriques, Ana Margarida; Caeiro, Raquel; Pista, Angela; Silveira, LeonorSalmonelose é a segunda zoonose mais frequente a nível mundial. É causada por Salmonella enterica, sendo S. Typhimurium um dos serotipos mais reportados na União Europeia. Um dos maiores riscos para a saúde pública é a presença de Salmonella spp. em alimentos, e consequente associação a surtos que podem atingir uma dimensão transfronteiriça. Este estudo teve como principal objetivo caraterizar fenotípica e genotipicamente isolados de Salmonella Typhimurium recebidos no Laboratório Nacional de Referência de Infeções Gastrintestinais do Instituto Nacional de Saúde Doutor Ricardo Jorge, na sequência do aumento considerável de isolados deste serotipo observado entre abril e maio de 2024. Foram estudados 83 isolados, dos quais 64 integravam o mesmo cluster genético (77,1%), Cluster 1, identificado por core genome multilocus sequence typing (Enterobase cgMLST V2 + HierCC V1). Todos os isolados pertenciam à sequência tipo 19. Foi identificada a presença dos genes de resistência blaCARB-2, sul1, aadA2, aac(6’)-Iaa, tet(G) e floR em 67 isolados, perfil concordante com o fenótipo encontrado, AMP-AMC-TET-CHL. A análise de cgMLST permitiu ainda identificar quatro isolados europeus que integravam o Cluster 1.
- Budget impact analysis of cervical cancer screening in Portugal: comparison of cytology and primary HPV screening strategiesPublication . Pista, Angela; Costa, Carlos; Saldanha, Conceição; Moutinho, José Alberto Fonseca; Moutinho, José Maria; Arrobas, Fernando; Catalão, Carlos; Kempers, JariBackground: Primary Human Papilloma Virus (HPV) testing is the currently recommended cervical cancer (CxCa) screening strategy by the Portuguese Society of Gynecology (SPG) clinical consensus. However, primary HPV testing has not yet been adopted by the Portuguese organized screening programs. This modelling study compares clinical benefits and costs of replacing the current practice, namely cytology with ASCUS HPV triage, with 2 comparative strategies: 1) HPV (pooled) test with cytology triage, or 2) HPV test with 16/18 genotyping and cytology triage, in organized CxCa screenings in Portugal. Methods: A budget impact model compares screening performance, clinical outcomes and budget impact of the 3 screening strategies. A hypothetical cohort of 2,078,039 Portuguese women aged 25-64 years old women is followed for two screening cycles. Screening intervals are 3 years for cytology and 5 years for the HPV strategies. Model inputs include epidemiological, test performance and medical cost data. Clinical impacts are assessed with the numbers of CIN2-3 and CxCa detected. Annual costs, budget impact and cost of detecting one CIN2+ were calculated from a public healthcare payer's perspective. Results: HPV testing with HPV16/18 genotyping and cytology triage (comparator 2) shows the best clinical outcomes at the same cost as comparator 1 and is the most cost-effective CxCa screening strategy in the Portuguese context. Compared to screening with cytology, it would reduce annual CxCa incidence from 9.3 to 5.3 per 100,000, and CxCa mortality from 2.7 to 1.1 per 100,000. Further, it generates substantial cost savings by reducing the annual costs by €9.16 million (- 24%). The cost of detecting CIN2+ decreases from the current €15,845 to €12,795. On the other hand, HPV (pooled) test with cytology triage (comparator 1) reduces annual incidence of CxCa to 6.9 per 100,000 and CxCa mortality to 1.6 per 100,000, with a cost of €13,227 per CIN2+ detected with annual savings of €9.36 million (- 24%). The savings are mainly caused by increasing the length of routine screening intervals from three to five years. Conclusion: The results support current clinical recommendations to replace cytology with HPV with 16/18 genotyping with cytology triage as screening algorithm.
- Building an International One Health Strain Level Database to Characterise the Epidemiology of AMR Threats: ESBL—AmpC Producing E. coli as An Example—Challenges and PerspectivesPublication . Perestrelo, Sara; Amaro, Ana; Brouwer, Michael S.M.; Clemente, Lurdes; Ribeiro Duarte, Ana Sofia; Kaesbohrer, Annemarie; Karpíšková, Renata; Lopez-Chavarrias, Vicente; Morris, Dearbháile; Prendergast, Deirdre; Pista, Angela; Silveira, Leonor; Skarżyńska, Magdalena; Slowey, Rosemarie; Veldman, Kees T.; Zając, Magdalena; Burgess, Catherine; Alvarez, JulioAntimicrobial resistance (AMR) is one of the top public health threats nowadays. Among the most important AMR pathogens, Escherichia coli resistant to extended spectrum cephalosporins (ESC-EC) is a perfect example of the One Health problem due to its global distribution in animal, human, and environmental sources and its resistant phenotype, derived from the carriage of plasmid-borne extended-spectrum and AmpC β-lactamases, which limits the choice of effective antimicrobial therapies. The epidemiology of ESC-EC infection is complex as a result of the multiple possible sources involved in its transmission, and its study would require databases ideally comprising information from animal (livestock, companion, wildlife), human, and environmental sources. Here, we present the steps taken to assemble a database with phenotypic and genetic information on 10,763 ESC-EC isolates retrieved from multiple sources provided by 13 partners located in eight European countries, in the frame of the DiSCoVeR Joint Research project funded by the One Health European Joint Programme (OH-EJP), along with its strengths and limitations. This database represents a first step to help in the assessment of different geographical and temporal trends and transmission dynamics in animals and humans. The work performed highlights aspects that should be considered in future international efforts, such as the one presented here.
- Caracterização fenotípica e genotípica de isolados de Salmonella Typhi e Paratyphi em Portugal, entre janeiro de 1994 e maio de 2024Publication . Henriques, Ana Margarida; Caeiro, Raquel; Tomás, Alexandra; Silveira, Leonor; Pista, AngelaOs serotipos Typhi e Paratyphi causam infeções sistémicas graves, sendo crucial a administração de antibioterapia adequada. No entanto, o aumento crescente da taxa de multirresistência aos antibióticos, especialmente no caso de Salmonella Typhi, dificulta a escolha dos antibióticos a administrar. Em Portugal, os casos de febre tifoide e paratifoide são raros. Com este estudo pretendeu-se identificar e caracterizar os perfis de resistência aos antibióticos de isolados de Salmonella Typhi e Salmonella Paratyphi A, B e C, entre janeiro de 1994 e maio de 2024, e avaliar a sua proximidade genética. Após confirmação do serotipo, 86 isolados foram submetidos a testes de suscetibilidade aos antibióticos e sequenciação total do genoma. Neste estudo, 60,5% dos isolados foram resistentes a pelo menos um antibiótico, dos quais 15,4% eram multirresistentes. Entre 1994-2016 todos os isolados foram suscetíveis, mas a partir de 2017 foi possível observar uma tendência crescente na resistência aos antibióticos, especialmente para S. Typhi. A resistência à pefloxacina foi a mais frequente, seguida da resistência ao ácido nalidíxico, ampicilina, trimetoprim, sulfametoxazol e cloranfenicol. Dos diversos genes de resistência identificados, destaca-se a presença do gene blaCTX- M-15 num isolado de Salmonella Typhi de 2020, resistente a antibióticos de importância crítica. Foram identificados 11 pequenos clusters. Apesar dos serotipos tifoides e paratifoides não serem endémicos em Portugal, é essencial manter uma vigilância laboratorial contínua, de forma a caracterizar os isolados circulantes em Portugal e detetar potenciais surtos.
- Caracterização genómica de Salmonella Newport: identificação de clusters genéticos, janeiro 1991-maio 2024Publication . Caeiro, Raquel; Tomás, Alexandra; Henriques, Ana Margarida; Pista, Angela; Silveira, LeonorSalmonella enterica subsp. enterica serovar Newport (S. Newport) é um serotipo pouco frequente na União Europeia. No entanto, devido à sua associação com alguns surtos de origem alimentar, a sua importância não deve ser negligenciada. Em Portugal, os dados sobre a infeção por S. Newport são escassos. Este estudo teve como principais objetivos caracterizar fenotípica e genotipicamente isolados clínicos de S. Newport entre 1991 e maio de 2024, e identificar possíveis clusters genéticos. Após serotipagem, 98 isolados foram submetidos a testes de suscetibilidade aos antibióticos e sequenciação total do genoma. Apesar da maioria dos isolados estudados (96,9%) terem sido suscetíveis aos antibióticos testados, em 2024 foi identificado um isolado multirresistente, que, entre outros, continha o gene blaDHA-1. A análise core genome multilocus sequence typing (Enterobase cgMLST V2 + HierCC V1), considerando um threshold de cinco diferenças alélicas, permitiu identificar oito clusters distintos, dois dos quais de maiores dimensões: o Cluster 1, com 32 isolados de 2019, e o Cluster 2, com quatro isolados de 2022 e 18 de 2023. Embora S. Newport seja um serotipo pouco frequente em Portugal e não apresentar, na maioria dos casos, resistências aos antibióticos, é essencial manter uma vigilância laboratorial contínua, que permita de forma atempada implementar medidas de prevenção e controlo eficazes.
- Clinical Performance of the CLART Human Papillomavirus 2 Assay Compared With the Hybrid Capture 2 TestPublication . Pista, Angela; Verdasca, Nuno; Oliveira, AnaPersistent infection by high-risk human papillomavirus (HR-HPV) is a cause of cervical cancer. The use of HPV detection in cervical screening programs may improve the ability to identify women at risk of cervical cancer. Therefore, the development of appropriate methods for the detection of HR-HPV is essential. The aim of this study was to evaluate the clinical performance of the CLART Human Papillomavirus 2 assay (CLART) in comparison with the Hybrid Capture 2 test (HC2), using a clinical cut-off of cervical intraepithelial neoplasia grade 2 or worse. Discrepant results were analyzed further by the PapilloCheck HPV genotyping system. In the 425 studied women, HR-HPV positivity rates were similar by both tests (CLART-13 HR-HPV: 63.1%; CLART-17 HR-HPV: 64.7%; HC2: 64.5%). Agreement between CLART-13 HR-HPV (k¼ 0.969; concordance level 98.6%), CLART-17 HR-HPV (k¼ 0.974; concordance level 98.8%), and HC2 were very good. When 13 HR-HPV types were considered, the two tests showed a clinical sensitivity of 96% (95% CI: 92.6–97.9). The clinical specificity of CLART-13 HR-HPV was 73.6% (95% CI: 66.7–79.5) for cervical intraepithelial neoplasia grade 2 or worse, which was comparable to HC2 (71.4%; 95% CI: 64.3–77.5). When all 17 HR-HPV types were considered, CLART showed a clinical sensitivity of 96.9% (95% CI: 93.8–98.5) and a clinical specificity of 71.9% (95% CI: 64.9–78.0). In conclusion, the CLART assay is efficient, sensitive, reproducible, and has a similar performance to HC2 for cervical intraepithelial neoplasia grade 2 or worse. Furthermore, this assay has the advantage of detecting and genotyping 35 HPV types by a single test, which can provide additional information on the predictive value of infection with HR-HPV
- Genetic Diversity of Salmonella enterica subsp. enterica Serovar Enteritidis from Human and Non-Human Sources in PortugalPublication . Leão, Célia; Silveira, Leonor; Usié, Ana; Gião, Joana; Clemente, Lurdes; Themudo, Patricia; Amaro, Ana; Pista, AngelaSalmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is one of the leading causes of foodborne infections associated with broilers and laying hens. Portugal has had the lowest notification rates of salmonellosis in recent years, due to the vaccinations of layer and breeder flocks and strict compliance with biosecurity measures. However, data about the genetic diversity of S. Enteritidis in Portugal are scarce. In this study, 102 S. Enteritidis isolates selected from human (n = 63) and non-human sources (n = 39) were characterized by serotyping, antimicrobial susceptibility, and whole genome sequencing. The S. Enteritidis population was mainly resistant to fluoroquinolones, and a sole isolate showed resistance to extended-spectrum cephalosporins. ST11 was the most frequent sequence type, and three novel STs from human isolates (ST9236, ST4457, and ST9995) were assigned. Several Salmonella pathogenic islands (SPI) and Putative SPI were present in the genomes, namely SPI-1, 2, 3, 4, 5, 9, 10, 12, 13, and 14, C63PI, CS54_island, and 170 virulence genes were identified. The phylogenetic analysis revealed that strains from Portugal are genetically heterogeneous regarding sample type, collection date, and genetic content. This study increases the available data, essential to a better characterization of strains in a global context.
- Genotypic and Phenotypic Characterization of Pathogenic Escherichia coli, Salmonella spp., and Campylobacter spp., in Free-Living Birds in Mainland PortugalPublication . Batista, Rita; Saraiva, Margarida; Lopes, Teresa; Silveira, Leonor; Coelho, Anabela; Furtado, Rosália; Castro, Rita; Correia, Cristina Belo; Rodrigues, David; Henriques, Pedro; Lóio, Sara; Soeiro, Vanessa; da Costa, Paulo Martins; Oleastro, Mónica; Pista, AngelaBirds are potential carriers of pathogens affecting humans and agriculture. Aiming to evaluate the occurrence of the top three most important foodborne pathogens in free-living birds in Portugal, we investigated 108 individual fecal samples from free-living birds and one pooled sample of gull feces (n = 50) for the presence of Escherichia coli (pathogenic and non-pathogenic), Salmonella spp. and Campylobacter spp. Virulence- and antimicrobial resistance- (AMR) associated genes were detected by PCR and Whole-Genome Sequencing (WGS), and phenotypic (serotyping and AMR profiles) characterization was performed. Overall, 8.9% of samples tested positive for pathogenic E. coli, 2.8% for Salmonella spp., and 9.9% for Campylobacter spp. AMR was performed on all pathogenic isolates and in a fraction of non-pathogenic E. coli, being detected in 25.9% of them. Ten of the tested E. coli isolates were multidrug-resistant (MDR), and seven of them were Extended-spectrum β-lactamase (ESBL) producers. Among Salmonella (n = 3) and Campylobacter (n = 9), only one strain of C. jejuni was identified as MDR. Most of the identified serotypes/sequence types had already been found to be associated with human disease. These results show that free-living birds in Portugal may act as carriers of foodborne pathogens linked to human disease, some of them resistant to critically important antimicrobials.
- HPV – O Cancro do Colo do ÚteroPublication . Pista, Angela
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