Browsing by Author "de Carvalho, Isabel Lopes"
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- An Overview of Monkeypox Virus Detection in Different Clinical Samples and Analysis of Temporal Viral Load DynamicsPublication . Cordeiro, Rita; Pelerito, Ana; de Carvalho, Isabel Lopes; Lopo, Sílvia; Neves, Raquel; Rocha, Raquel; Palminha, Paula; Verdasca, Nuno; Palhinhas, Cláudia; Borrego, Maria José; Manita, Carla; Ferreira, Idalina; Bettencourt, Célia; Vieira, Patrícia; Silva, Sónia; Água-Doce, Ivone; Roque, Carla; Cordeiro, Dora; Brondani, Greice; Santos, João Almeida; Martins, Susana; Rodrigues, Irene; Ribeiro, Carlos; Núncio, Maria Sofia; Gomes, João Paulo; Batista, Fernando da ConceiçãoMpox is a zoonotic disease caused by the Monkeypox virus (MPXV), and since May 2022, tens of thousands of cases have been reported in non-endemic countries. We aimed to evaluate the suitability of different sample types for mpox diagnostic and assess the temporal dynamics of viral load. We evaluated 1914 samples from 953 laboratory-confirmed cases. The positivity rate was higher for lesion (91.3%) and rectal swabs (86.1%) when compared with oropharyngeal swabs (69.5%) and urines (41.2%), indicating higher viral loads for the former. Supporting this, lesion and rectal swabs showed lower median PCR C values (C = 23 and C = 24), compared to oropharyngeal swabs and urines (C = 31). Stable MPXV loads were observed in swabs from lesions up to 30 days after symptoms onset, contrasting with a considerable decrease in viral load in rectal and oropharyngeal swabs. Overall, these results point to lesion swabs as the most suitable samples for detecting MPXV in the 2022-2023 multicountry outbreak and show comparable accuracy to rectal swabs up to 8 days after symptoms onset. These findings, together with the observation that about 5% of patients were diagnosed through oropharyngeal swabs while having negative lesions, suggest that multisite testing should be performed to increase diagnostic sensitivity.
- Borrelia garinii and Francisella tularensis subsp. holarctica detected in migratory shorebirds in PortugalPublication . de Carvalho, Isabel Lopes; Alves, Ana Sofia; Pardal, Sara; Lopes, Ricardo Jorge; Mendes, Luísa; Núncio, Maria Sofia; Zé-Zé, LíbiaMigratory shorebirds use, among many, the East Atlantic Flyway that links breeding areas as north as Tundra habitats to aquatic wintering grounds in West Africa. As a consequence, they are potentially important in the spread of global zoonotic diseases transmitted by ticks, such as Lyme borreliosis and tularemia—two diseases previously detected in Portugal. In this study, we looked at the infection status of seven populations of shorebirds during their migration, breeding, or wintering in the Portuguese wetlands to access if they carry these pathogens and to discuss their potential risk in the Portuguese wetlands. A total of 212 migratory shorebirds captured in the Tagus and Sado estuaries; key staging and wintering sites in this flyway and important breeding areas for some species were analyzed for the presence of Borrelia burgdorferi sensu lato and Francisella tularensis. In the present study, B. garinii was identified in seven (3%) specimens (five black-tailed godwits Limosa limosa, one common redshank Tringa totanus, and one little stint Calidris minuta), whereas F. tularensis subsp. holarctica was identified in one (0.4%) little stint. To our knowledge, this is the first evidence that shorebirds that migrate through or winter in Portugal transport these pathogens, potentially contributing for their introduction along the flyway, including the Mediterranean region.
- Molecular characterization of a new isolate of Borrelia lusitaniae derived from Apodemus sylvaticus in PortugalPublication . de Carvalho, Isabel Lopes; Zeidner, N.; Ullmann, A.; Hojgaard, A.; Amaro, F.; Zé-Zé, Líbia; Alves, M.J.; De Sousa, R.; Piesman, J.; Núncio, M.S.A total of 196 small mammals were collected in Portugal and tested for Borrelia burgdorferi sensu lato. Tissue samples were taken from each animal and cultured in Barbour-Stoenner-Kelly (BSK)-II medium. The single strain of spirochete isolated was confirmed as Borrelia lusitaniae by genetic analyses. This is the first report of B. lusitaniae isolated from Apodemus sylvaticus.
- Natural selection and recombination at host-interacting lipoprotein loci drive genome diversification of Lyme disease and related bacteriaPublication . Akther, Saymon; Mongodin, Emmanuel F.; Morgan, Richard D.; Di, Lia; Yang, Xiaohua; Golovchenko, Maryna; Rudenko, Natalie; Margos, Gabriele; Hepner, Sabrina; Fingerle, Volker; Kawabata, Hiroki; Norte, Ana Cláudia; de Carvalho, Isabel Lopes; Núncio, Maria Sofia; Marques, Adriana; Schutzer, Steven E.; Fraser, Claire M.; Luft, Benjamin J.; Casjens, Sherwood R.; Qiu, WeigangLyme disease, caused by spirochetes in the Borrelia burgdorferi sensu lato clade within the Borrelia genus, is transmitted by Ixodes ticks and is currently the most prevalent and rapidly expanding tick-borne disease in Europe and North America. We report complete genome sequences of 47 isolates that encompass all established species in this clade while highlighting the diversity of the widespread human pathogenic species B. burgdorferi. A similar set of plasmids has been maintained throughout Borrelia divergence, indicating that they are a key adaptive feature of this genus. Phylogenetic reconstruction of all sequenced Borrelia genomes revealed the original divergence of Eurasian and North American lineages and subsequent dispersals that introduced B. garinii, B. bavariensis, B. lusitaniae, B. valaisiana, and B. afzelii from East Asia to Europe and B. burgdorferi and B. finlandensis from North America to Europe. Molecular phylogenies of the universally present core replicons (chromosome and cp26 and lp54 plasmids) are highly consistent, revealing a strong clonal structure. Nonetheless, numerous inconsistencies between the genome and gene phylogenies indicate species dispersal, genetic exchanges, and rapid sequence evolution at plasmid-borne loci, including key host-interacting lipoprotein genes. While localized recombination occurs uniformly on the main chromosome at a rate comparable to mutation, lipoprotein-encoding loci are recombination hotspots on the plasmids, suggesting adaptive maintenance of recombinant alleles at loci directly interacting with the host. We conclude that within- and between-species recombination facilitates adaptive sequence evolution of host-interacting lipoprotein loci and contributes to human virulence despite a genome-wide clonal structure of its natural populations. Importance: Lyme disease (also called Lyme borreliosis in Europe), a condition caused by spirochete bacteria of the genus Borrelia, transmitted by hard-bodied Ixodes ticks, is currently the most prevalent and rapidly expanding tick-borne disease in the United States and Europe. Borrelia interspecies and intraspecies genome comparisons of Lyme disease-related bacteria are essential to reconstruct their evolutionary origins, track epidemiological spread, identify molecular mechanisms of human pathogenicity, and design molecular and ecological approaches to disease prevention, diagnosis, and treatment. These Lyme disease-associated bacteria harbor complex genomes that encode many genes that do not have homologs in other organisms and are distributed across multiple linear and circular plasmids. The functional significance of most of the plasmid-borne genes and the multipartite genome organization itself remains unknown. Here we sequenced, assembled, and analyzed whole genomes of 47 Borrelia isolates from around the world, including multiple isolates of the human pathogenic species. Our analysis elucidates the evolutionary origins, historical migration, and sources of genomic variability of these clinically important pathogens. We have developed web-based software tools (BorreliaBase.org) to facilitate dissemination and continued comparative analysis of Borrelia genomes to identify determinants of human pathogenicity.
- Phylogenomic characterization and signs of microevolution in the 2022 multi-country outbreak of monkeypox virusPublication . Isidro, Joana; Borges, Vítor; Pinto, Miguel; Sobral, Daniel; Santos, João Dourado; Nunes, Alexandra; Mixão, Verónica; Ferreira, Rita; Santos, Daniela; Duarte, Silvia; Vieira, Luís; Borrego, Maria José; Núncio, Sofia; de Carvalho, Isabel Lopes; Pelerito, Ana; Cordeiro, Rita; Gomes, João PauloThe largest monkeypox virus (MPXV) outbreak described so far in non-endemic countries was identified in May 2022 (refs. 1-6). In this study, shotgun metagenomics allowed the rapid reconstruction and phylogenomic characterization of the first MPXV outbreak genome sequences, showing that this MPXV belongs to clade 3 and that the outbreak most likely has a single origin. Although 2022 MPXV (lineage B.1) clustered with 2018-2019 cases linked to an endemic country, it segregates in a divergent phylogenetic branch, likely reflecting continuous accelerated evolution. An in-depth mutational analysis suggests the action of host APOBEC3 in viral evolution as well as signs of potential MPXV human adaptation in ongoing microevolution. Our findings also indicate that genome sequencing may provide resolution to track the spread and transmission of this presumably slow-evolving double-stranded DNA virus.
- Viral genetic clustering and transmission dynamics of the 2022 mpox outbreak in PortugalPublication . Borges, Vítor; Duque, Mariana Perez; Martins, João Vieira; Vasconcelos, Paula; Ferreira, Rita; Sobral, Daniel; Pelerito, Ana; de Carvalho, Isabel Lopes; Núncio, Maria Sofia; Borrego, Maria José; Roemer, Cornelius; Neher, Richard A.; O’Driscoll, Megan; Rocha, Raquel; Lopo, Sílvia; Neves, Raquel; Palminha, Paula; Coelho, Luís; Nunes, Alexandra; Isidro, Joana; Pinto, Miguel; Santos, João Dourado; Mixão, Verónica; Santos, Daniela; Duarte, Silvia; Vieira, Luís; Martins, Fátima; Machado, Jorge; Veríssimo, Vítor Cabral; Grau, Berta; Peralta-Santos, André; Neves, José; Caldeira, Margarida; Pestana, Mafalda; Fernandes, Cândida; Caria, João; Pinto, Raquel; Póvoas, Diana; Maltez, Fernando; Sá, Ana Isabel; Salvador, Mafalda Brito; Teófilo, Eugénio; Rocha, Miguel; Moneti, Virginia; Duque, Luis Miguel; e Silva, Francisco Ferreira; Baptista, Teresa; Vasconcelos, Joana; Casanova, Sara; Mansinho, Kamal; Alves, João Vaz; Alves, João; Silva, António; Alpalhão, Miguel; Brazão, Cláudia; Sousa, Diogo; Filipe, Paulo; Pacheco, Patrícia; Peruzzu, Francesca; de Jesus, Rita Patrocínio; Ferreira, Luís; Mendez, Josefina; Jordão, Sofia; Duarte, Frederico; Gonçalves, Maria João; Pena, Eduarda; Silva, Claúdio Nunes; Guimarães, André Rodrigues; Tavares, Margarida; Freitas, Graça; Cordeiro, Rita; Gomes, João PauloPathogen genome sequencing during epidemics enhances our ability to identify and understand suspected clusters and investigate their relationships. Here, we combine genomic and epidemiological data of the 2022 mpox outbreak to better understand early viral spread, diversification and transmission dynamics. By sequencing 52% of the confirmed cases in Portugal, we identified the mpox virus sublineages with the highest impact on case numbers and fitted them into a global context, finding evidence that several international sublineages probably emerged or spread early in Portugal. We estimated a 62% infection reporting rate and that 1.3% of the population of men who have sex with men in Portugal were infected. We infer the critical role played by sexual networks and superspreader gatherings, such as sauna attendance, in the dissemination of mpox virus. Overall, our findings highlight genomic epidemiology as a tool for the real-time monitoring and control of mpox epidemics, and can guide future vaccine policy in a highly susceptible population.
- Viral genetics and transmission dynamics in the second wave of mpox outbreak in Portugal and forecasting public health scenariosPublication . Cordeiro, Rita; Caetano, Constantino P.; Sobral, Daniel; Ferreira, Rita; Coelho, Luís; Pelerito, Ana; de Carvalho, Isabel Lopes; Namorado, Sónia; Loyens, Dinis B.; Mexia, Ricardo; Fernandes, Cândida; Neves, José Miguel; João, Ana Luísa; Rocha, Miguel; Duque, Luís Miguel; Correia, Inês; Baptista, Teresa; Brazão, Cláudia; Sousa, Diogo; Filipe, Paulo; Alpalhão, Miguel; Maltez, Fernando; Póvoas, Diana; Pinto, Raquel; Caria, João; Patrocínio de Jesus, Rita; Pacheco, Patrícia; Peruzzu, Francesca; Méndez, Josefina; Ferreira, Luís; Mansinho, Kamal; Alves, João Vaz; Vasconcelos, Joana; Domingos, João; Casanova, Sara; Duarte, Frederico; Gonçalves, Maria João; Salvador, Mafalda Brito; Guimarães, Mafalda Andresen; Martins, Sueila; Oliveira, Marvin Silva; Santos, Daniela; Vieira, Luís; Núncio, Maria Sofia; Borges, Vítor; Gomes, João PauloIn 2023, a second wave of the global mpox epidemic, which is mainly affecting men who have sex with men (MSM), was observed in some countries. Herein, we benefited from a large viral sequence sampling (76/121; 63%) and vast epidemiological data to characterise the re-emergence and circulation of the (MPXV) in Portugal during 2023. We also modelled transmission and forecasted public health scenarios through a compartmental susceptible-exposed-infectious-recovered (SEIR) model. Our results suggest that the 2023 mpox wave in Portugal resulted from limited introduction(s) of MPXV belonging to C.1.1 sublineage, hypothetically from Asia, followed by sustained viral transmission and potential exportation to other countries. We estimated that the contribution of the MSM high sexual activity group to mpox transmission was 120 (95% CrI: 30-3553) times higher than that of the low sexual activity group. However, among the high sexual activity group, vaccinated individuals likely contributed approximately eight times less [0.123 (95% CrI: 0.068-0.208)] than the unvaccinated ones. Vaccination was also linked to potential reduced disease severity, with a Mpox Severity Score of 6.0 in the vaccinated group compared to 7.0 in unvaccinated individuals. Scenario analysis indicated that transmission is highly sensitive to sexual behaviour, projecting that a slight increase in the MSM sub-population with high sexual activity can trigger new mpox waves. This study strongly supports that continued vaccination, targeted awareness among risk groups and routine genomic epidemiology is needed to anticipate and respond to novel MPXV threats (e.g. global dissemination of clade I viruses).