Ticks collected from humans and tick-borne agents within the National Network for Vector Surveillance (REVIVE), 2011-2015

A Rede de Vigilância de Vetores REVIVE – Ixodídeos foi desenvolvida em Portugal para vigiar e aumentar o conhecimento sobre as espécies de ixodídeos presentes e dos agentes patogénicos a estes associados. Esta rede foi estabelecida em 2011 apresentando este estudo os resultados obtidos nos primeiros cinco anos de vigilância nas carraças removidas de humanos, relativamente às espécies ixodológicas, sua abundância, sazonalidade e presença de Rickettsia e Borrelia.REVIVE - Ticks is a National Network for Vector Surveillance developed to enhance knowledge about ticks and the pathogens that they may carry. The Network was established in 2011 and this study presents the results obtained during five years of surveillance of ticks removed from humans concerning tick species, relative abundance, seasonality and the presence of Rickettsia and Borrelia.info:eu-repo/semantics/acceptedVersio

Francisella-like endosymbiont in Dermacentor reticulatus collected in Portugal

In Portugal, recent studies have confirmed the presence of Francisella tularensis in Dermacentor reticulatus. Bacterial endosymbionts with significant homology to F. tularensis have been described in several species of ticks. In this work we identified Francisella-like endosymbionts in D. reticulatus ticks (39%), confirming the presence of these bacteria in Portugal. This finding should be considered in future studies using molecular approaches to detect Francisella prevalence in ticks and environmental samples

First Case of Tularemia Reported in Portugal: Probably of Imported Origin

The authors report the case of a 47-year-old man who walked in the countryside on the island of Bornholm, during the summer period. Three days later, fever, myalgias and adynamia began. The serological tests, Real-time PCR and isolation of the bacteria from the culture of lymph biopsy confirmed the presence of Francisella tularensis subsp. holarctica

Portuguese hosts for Ornithodoros erraticus ticks

The hematophagous soft tick Ornithodoros erraticus feeds nocturnally on multiple warm-blooded vertebrate hosts. This tick is often found living buried in the soil of traditional pigpens. O. erraticus is an important infectious disease vector both for humans and animals. In the Iberian Peninsula, this tick serves as the vector of human tick-borne relapsing fever caused by the spirochete Borrelia hispanica. The natural ecosystems maintaining this spirochete are not well understood, with details of competent vertebrate reservoirs and tick–host interactions poorly understood. Investigation of arthropod blood meal composition provides evidence linking the vector to specific hosts, providing insights into possible disease reservoirs. Ticks collected from two pigpens located in southern Portugal were subjected to blood meal analysis. PCR amplification of vertebrate cytochrome b was used to disclose the original host from which 349 ticks had derived their previous blood meal. Host origins for blood meal analysis from 79 of 349 ticks revealed that 46.8% had previously fed from pigs, 35.4% human, 13.9% bovine, 5.1% sheep, 1.3% rodent, and 1.3% from birds. Three samples revealed mixed blood meals, namely, human–pig (1.3%), sheep–pig (1.3%), and bovine–pig (1.3%). The major role of pigs as hosts is consistent with fieldwork observations and underlines the importance of pigs for maintaining O. erraticus tick populations. Humans serve as accidental hosts, frequently confirmed by reports from both producers and veterinarians. Other livestock species and wildlife prevalent in the region appear only to have a minor role in maintaining this tick. The results demonstrate the importance of blood meal analysis to determine tick hosts providing a tool for investigation of sylvatic cycle for Borrelia hispanica

Portuguese Hosts for Ornithodoros erraticus Ticks

The hematophagous soft tick Ornithodoros erraticus feeds nocturnally on multiple warm-blooded vertebrate hosts. This tick is often found living buried in the soil of traditional pigpens. O. erraticus is an important infectious disease vector both for humans and animals. In the Iberian Peninsula, this tick serves as the vector of human tick-borne relapsing fever caused by the spirochete Borrelia hispanica. The natural ecosystems maintaining this spirochete are not well understood, with details of competent vertebrate reservoirs and tick–host interactions poorly understood. Investigation of arthropod blood meal composition provides evidence linking the vector to specific hosts, providing insights into possible disease reservoirs. Ticks collected from two pigpens located in southern Portugal were subjected to blood meal analysis. PCR amplification of vertebrate cytochrome b was used to disclose the original host from which 349 ticks had derived their previous blood meal. Host origins for blood meal analysis from 79 of 349 ticks revealed that 46.8% had previously fed from pigs, 35.4% human, 13.9% bovine, 5.1% sheep, 1.3% rodent, and 1.3% from birds. Three samples revealed mixed blood meals, namely, human–pig (1.3%), sheep–pig (1.3%), and bovine–pig (1.3%). The major role of pigs as hosts is consistent with fieldwork observations and underlines the importance of pigs for maintaining O. erraticus tick populations. Humans serve as accidental hosts, frequently confirmed by reports from both producers and veterinarians. Other livestock species and wildlife prevalent in the region appear only to have a minor role in maintaining this tick. The results demonstrate the importance of blood meal analysis to determine tick hosts providing a tool for investigation of sylvatic cycle for Borrelia hispanica

Vector Surveillance Network (REVIVE): ixodids and pathogenic bacteria detected in mainland Portugal during 2019

A Rede de Vigilância de Vetores (REVIVE) resulta da colaboração entre instituições do Ministério da Saúde (Direção-Geral da Saúde, Administrações Regionais de Saúde, Instituto dos Assuntos Sociais e da Saúde da Madeira, Direção Regional de Saúde dos Açores e Instituto Nacional de Saúde Doutor Ricardo Jorge). A Rede divide-se em duas vertentes, REVIVE-mosquitos e REVIVE-carraças. Neste artigo apresentam-se, de forma resumida, os resultados obtidos em 2019 no âmbito do REVIVE- -carraças. Este programa, que existe há nove anos, tem como principais objetivos monitorizar a atividade de artrópodes hematófagos; caracterizar as espécies e sua ocorrência sazonal; identificar agentes patogénicos importantes em saúde pública, a densidade dos vetores, o nível de infeção e monitorizar a introdução de espécies exóticas. Das atividades desenvolvidas em 2019, destaca-se: a participação das cinco Administrações Regionais de Saúde que realizaram colheitas de carraças em 166 concelhos; nos 2409 ixodídeos colhidos não foi identificada a presença de espécies exóticas; em 982 carraças foi pesquisada a presença de borrélias e rickettsias, tendo sido observada a prevalência média de 2% e 22%, respetivamente, sobretudo em carraças colhidas a parasitar humanos. O REVIVE-carraças continua a contribuir para um conhecimento sistemático da fauna de ixodídeos de Portugal, e do seu potencial papel de vetor, constituindo uma componente dos programas de vigilância epidemiológica indispensável à avaliação do risco de transmissão de doenças potencialmente graves.The Vector Surveillance Network (REVIVE) results from the collaboration between institutions of the Ministry of Health (Directorate-General for Health, Regional Health Administrations, Institute of Social Affairs and Health of Madeira, Regional Directorate of Health of the Azores and National Institute of Health Dr Ricardo Jorge). It is divided into two areas, REVIVE-mosquitoes and REVIVE-ticks. In this article, the results obtained in 2019 in the context of REVIVE-ticks are summarized. This program, which has completed nine years, aims to monitor the activity of blood-sucking arthropods; to characterize the species and their seasonal occurrence; to identify important pathogens in public health, the density of the vectors, the level of infection and monitor the introduction of exotic species. Among the activities developed in 2019 stands out: the participation of the five Regional Health Administrations that carried out tick harvesting in 166 municipalities; in the 2409 ixodids collected, the presence of exotic species was not identified; in 982 ticks, the presence of borrelias and rickettsiae was investigated, with an average prevalence of 2% and 22%, respectively, especially in ticks harvested when parasitizing humans. REVIVE-ticks continues to contribute to a systematic knowledge of the fauna of ixodids in Portugal, and its potential role as a vector, constituting a component of epidemiological surveillance programs indispensable to assess the risk of transmission of potentially serious diseases.info:eu-repo/semantics/publishedVersio

Whole Genome Sequencing-Based Approach for the Determination of Multiple Locus Variable Number Tandem Repeat Profiles

Funding Information: Funding: This work is a result of the GenomePT project (POCI-01- 0145-FEDER-022184), supported by the COMPETE 2020 – Operational Program for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Program (Lisboa2020), Algarve Portugal Regional Operational Program (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by the Fundação para a Ciência e a Tecnologia (FCT). The studies have arisen from the Project QUANDHIP (Chafea Grant Agreement no. 2010 21 02), which has been funded by the European Commission in the framework of the Health Program.Brucellosis is an important zoonosis that is emerging in some regions of the world, gaining increased relevance with the inclusion of the causing agent Brucella spp. in the class B bioterrorism group. Until now, multi-locus VNTR Analysis (MLVA) based on 16 loci has been considered as the gold standard for Brucella typing. However, this methodology is laborious, and, with the rampant release of Brucella genomes, the transition from the traditional MLVA to whole genome sequencing (WGS)-based typing is on course. Nevertheless, in order to avoid a disruptive transition with the loss of massive genetic data obtained throughout the last decade and considering that the transition timings will vary considerably among different countries, it is important to determine WGS-based MLVA alleles of the nowadays sequenced genomes. On this regard, we aimed to evaluate the performance of a Python script that had been previously developed for the rapid in silico extraction of the MLVA alleles, by comparing it to the PCR-based MLVA procedure over 83 strains from different Brucella species. The WGS-based MLVA approach detected 95.3% of all possible 1,328 hits (83 strains×16 loci) and showed an agreement rate with the PCR-based MLVA procedure of 96.4% for MLVA-16. According to our dataset, we suggest the use of a minimal depth of coverage of ~50x and a maximum number of ~200 contigs as guiding “boundaries” for the future application of the script. In conclusion, the evaluated script seems to be a very useful and robust tool for the in silico determination of MLVA profiles of Brucella strains, allowing retrospective and prospective molecular epidemiological studies, which are important for maintaining an active epidemiological surveillance of brucellosis.publishersversionpublishe

Phylogenomic characterization and signs of microevolution in the 2022 multi-country outbreak of monkeypox virus

Erratum: Nat Med. 2022 Oct;28(10):2220-2221. doi: 10.1038/s41591-022-02036-2. https://www.nature.com/articles/s41591-022-02036-2The 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.info:eu-repo/semantics/publishedVersio