Co-Circulation of Leishmania Parasites and Phleboviruses in a Population of Sand Flies Collected in the South of Portugal

This article belongs to the Special Issue Leishmaniasis: Vector-Host-Pathogen Interactions in Health and Disease.In the Old World, phlebotomine sand flies from the genus Phlebotomus are implicated in the transmission of Leishmania spp. parasites (Kinetoplastida: Trypanosomatidae) and viruses belonging to the genus Phlebovirus (Bunyavirales: Phenuiviridae). Two of the five sand fly species known to occur in Portugal, Phlebotomus perniciosus and Ph. ariasi, the former being the most ubiquitous, are recognized vectors of Leishmania infantum, which causes visceral leishmaniasis, the most prevalent form of leishmaniasis in the country. Phlebotomus perniciosus is also the vector of the neurotropic Toscana virus, which can cause aseptic meningitis. Entomological surveillance is essential to provide fundamental data about the presence of vectors and the pathogens they can carry. As such, and given the lack of data in Portugal, an entomological survey took place in the Algarve, the southernmost region of the country, from May to October 2018. Polymerase chain reaction assays were performed in order to detect the presence of the above-mentioned pathogens in sand fly pools. Not only were both Leishmania parasites and phleboviruses detected during this study, but more importantly, it was the first time their co-circulation was verified in the same sand fly population collected in Portugal.This research was partially supported by the Portuguese Foundation for Science and Technology (FCT) through the project: “Phleboviruses in Portugal—vectors, pathogenesis and co-infections” (PTDC/DTP-SAP/0859/2014).info:eu-repo/semantics/publishedVersio

Chicken feather keratin peptides for the control of keratinocyte migration

FAO estimates that in 2030 the poultry meat production could reach 120 million tons, which is a challenge in terms of waste management. Feathers are mainly composed of keratin, an important biomaterial. Using feathers as a source of keratin will minimize the waste generated, while contributing to supply an important material for several industries, such as pharmaceutical and biomedical. The peptides were extracted from the feathers by microbial degradation. In this study, we evaluated the peptides effect on keratinocyte metabolic activity and migration. The influence of these peptides on non-activated and activated macrophages was also assessed. It was demonstrated that depending on the keratin peptide fraction in contact with keratinocytes, it is possible to modulate the migration rate of the keratinocytes. Peptide fraction with low molecular weight increases migration, while peptides with a high range of molecular sizes decreases it. Some peptide fractions induce the secretion of TNF- in non-activated macrophages and not on activated macrophages, demonstrating that these peptides should only be placed in contact with cells, in the context of an ongoing inflammatory process. This work is a step forward on the understanding of keratin peptides influence on keratinocytes and immune cells system cells, macrophages.This research was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit. It was also was financed by the Portuguese Mass Spectrometry Network, integrated in the National Roadmap of Research Infrastructures of Strategic Relevance (ROTEIRO/0028/2013; LISBOA-01-0145-FEDER-022125) and the MAC 2014-2020-MAC/1.1b/042-INTERREG V A España-Portugal, “BIOTRANSFER 2: Transferencia de la investigación biotecnológica orientada a rentabilidad empresarial y movilización de flujos de negocio 2”.info:eu-repo/semantics/publishedVersio

Monitoring of the invasive mosquito Aedes albopictus in Portugal by the Vector Surveillance Network (REVIVE), 2017-2019

O mosquito Aedes albopictus, conhecido vulgarmente como Mos2017-2019quito Tigre Asiático, é um mosquito invasor procedente da região Ásia-Pacífico. Esta espécie é importante em saúde pública e veterinária devido ao seu papel na transmissão de diversos agentes patogénicos, nomeadamente os arbovírus chikungunya, dengue e Zika. Introduzido e estabelecido ao longo da costa mediterrânica europeia, incluindo Espanha desde 2004, este mosquito vetor tem sido responsável pelos surtos autóctones de arbovírus que têm ocorrido na Europa desde 2007. Em Portugal, o Ae. albopictus foi detetado pela primeira vez no âmbito da Rede de Vigilância de Vetores (REVIVE) em dois eventos de introdução independentes, na região norte em 2017 e no Algarve em 2018. O presente estudo teve como objetivo caracterizar a distribuição geográfica e atividade sazonal da espécie invasora Ae. albopictus em Portugal. A vigilância ativa foi significativamente incrementada pela REVIVE após estas deteções. Foram utilizadas armadilhas específicas para mosquitos adultos (Biogents Sentinel traps) e para as fases imaturas (Ovitraps) para determinar a sua distribuição geográfica e sazonal, abundância e avaliar o processo de estabelecimento e dispersão da espécie a nível regional e nacional. Estes dados são indispensáveis no planeamento estratégico de medidas de controlo vetorial para evitar a dispersão das populações de mosquitos vetores e prevenir surtos de doenças associadas a Ae. albopictus.The mosquito Aedes albopictus, also known as the Asian Tiger Mosquito, is an invasive mosquito from the Asia-Pacific region. This species is important in public and veterinary health due to its role in the transmission of several pathogens, namely the arboviruses chikungunya, dengue and Zika. Introduced and established along the European Mediterranean coast, including Spain since 2004, this vector mosquito has been responsible for the autochthonous arboviruses outbreaks that have been occurring more and more frequently in the European region since 2007. In Portugal, Ae. albopictus was detected for the first time by the Vector Surveillance Network (REVIVE) in two independent introduction events, in the northern region in 2017 and Algarve in 2018. The aim of the present study was to characterize the geographic distribution and seasonal activity of the invasive species Ae. albopictus in Portugal. Active surveillance was significantly increased by the REVIVE after these detections. Specific traps were used for adult mosquitoes (Biogents Sentinel traps) and for the immature stages (ovitraps) to determine its geographical and seasonal distribution, abundance and evaluate its way of establishment and dispersion at a regional and national level. These data are necessary in the strategic planning of vector control measures to avoid the dispersion of these populations of vectors and prevent outbreaks of vector-borne diseases.info:eu-repo/semantics/publishedVersio

OMICs approaches in diarrhetic shellfish toxins research

ReviewDiarrhetic shellfish toxins (DSTs) are among the most prevalent marine toxins in Europe’s and in other temperate coastal regions. These toxins are produced by several dinoflagellate species; however, the contamination of the marine trophic chain is often attributed to species of the genus Dinophysis. This group of toxins, constituted by okadaic acid (OA) and analogous molecules (dinophysistoxins, DTXs), are highly harmful to humans, causing severe poisoning symptoms caused by the ingestion of contaminated seafood. Knowledge on the mode of action and toxicology of OA and the chemical characterization and accumulation of DSTs in seafood species (bivalves, gastropods and crustaceans) has significantly contributed to understand the impacts of these toxins in humans. Considerable information is however missing, particularly at the molecular and metabolic levels involving toxin uptake, distribution, compartmentalization and biotransformation and the interaction of DSTs with aquatic organisms. Recent contributions to the knowledge of DSTs arise from transcriptomics and proteomics research. Indeed, OMICs constitute a research field dedicated to the systematic analysis on the organisms’ metabolisms. The methodologies used in OMICs are also highly e ective to identify critical metabolic pathways a ecting the physiology of the organisms. In this review, we analyze the main contributions provided so far by OMICs to DSTs research and discuss the prospects of OMICs with regard to the DSTs toxicology and the significance of these toxins to public health, food safety and aquacultureinfo:eu-repo/semantics/publishedVersio

Cardiac Amyloidosis Associated with Apolipoprotein A-IV Deposition Diagnosed by Mass Spectrometry-Based Proteomic Analysis

Amyloidosis is a group of disorders characterised by the accumulation of extracellular deposits of insoluble protein aggregates. Clinical management depends on the accurate identification of the amyloid precursor and underlying cause. We describe a rare case of apolipoprotein A-IV cardiac amyloidosis, the diagnosis of which required mass spectrometry-based proteomic analysis

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named “AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108”. AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species

West nile virus in Portugal: vector population, host interaction and detection of new flaviviruses

This thesis describes the studies undertaken in the epidemiology of West Nile virus (WNV) in Portugal exploring several key components of its natural cycle: identification of mosquito vector-populations – geographical and seasonal distribution; analysis of genetic and environmental determinants on feeding patterns of mosquito vectors; experimental transmission of WNV; identification of flaviviruses detected in wild mosquitoes; serological survey of wild birds potentially involved in the epizootic cycle of WNV; finally, a description of a clinical human case of WNV infection reported in 2010. Culex pipiens was one of the most abundant mosquito species demonstrating ecological and intrinsic competence to transmit WNV. No positive mosquito pools for WNV were detected, but new mosquito-only flaviviruses were identified. The presence of antibodies anti-WNV in a juvenile turtle-dove suggests local virus circulation. In conclusion, Portugal meets suitable conditions for epizootic circulation of WNV and for the occurrence of accidental human infections; RESUMO O vírus West Nile em Portugal Vetores, interação com hospedeiros e deteção de novos flavivírus Esta tese descreve os estudos desenvolvidos na epidemiologia do vírus West Nile (VWN) em Portugal explorando várias componentes do seu ciclo natural: identificação das populações de mosquitos vetores – distribuição geográfica/ sazonal; análise de determinantes genéticos/ ambientais na preferência de hospedeiro de mosquitos vetores; transmissão experimental do VWN; identificação de flavivírus detectados em mosquitos selvagens; estudo serológico em aves potencialmente reservatórios do vírus; finalmente é descrito um caso clínico humano de infecção por VWN detetado em 2010. Culex pipiens foi uma das espécies de mosquito mais abundantes e demonstrou competência ecológica e intrínseca para transmitir o VWN. Não foram detectados pools de mosquitos positivos para o VWN, mas foram identificados novos flavivírus específicos de insetos. A presença de anticorpos anti-WNV numa rola juvenil sugere circulação local do vírus. Em conclusão, Portugal reúne condições adequadas para a circulação epizoótica do VWN e para a ocorrência de infecções humanas acidentais

Automated and remote surveillance of mosquitoes in the National Vector Surveillance Network (REVIVE): implementation of traps with a new optical sensor system for automatic classification of mosquitoes

No âmbito da Rede de Vigilância de Vetores (REVIVE ), é realizada a vigilância entomológica em mosquitos desde 2008 em Portugal. O projeto REVIVE é uma componente dos programas de vigilância epidemiológica indispensável à avaliação do risco de transmissão de agentes patogénicos transmitidos por vetores e é fundamental para a implementação de uma estratégia integrada de controlo de vetores e prevenção de doenças. As atividades previstas nos programas de vigilância entomológica e doenças associadas exigem elevados custos com recursos humanos especializados e com a logística que é investida nas inspeções periódicas das armadilhas de insetos. É essencial o desenvolvimento de estratégias sustentáveis para a monitorização de vetores. O projeto VECTRACK propõe o desenvolvimento de um método inovador na vigilância de mosquitos vetores de agentes de doença que vai possibilitar a identificação da espécie, sexo, idade e potencial de infeção das populações de mosquitos, mas de forma automatizada e remota. Neste artigo apresentamos os principais protocolos usados no projeto VECTRACK para a configuração de um sensor acoplado a uma armadilha que proceda à identificação automatizada de mosquitos através de machine learning.Within the scope of the Vector Surveillance Network (REVIVE ), entomological surveillance of mosquitoes has been carried out since 2008 in Portugal. REVIVE project is an essential component of epidemiological surveillance programs to assess the risk of transmission of vector-borne pathogens and is essential for the implementation of an integrated strategy for vector control and disease prevention. The activities foreseen in the entomological surveillance programs and associated diseases demand high costs with specialized human resources and with the logistics that are invested in the periodic inspections of insect traps. The development of sustainable strategies for vector monitoring is essential. The VECTRACK project proposes the development of an innovative method in the surveillance of mosquito vectors of disease agents that will make it possible to identify the species, sex, age and infection potential of mosquito populations, but in an automated and remote way. In this manuscript, we present the main protocols used in the VECTRACK project for the configuration of a sensor coupled to a trap that performs the automated identification of mosquitoes through machine learning.info:eu-repo/semantics/publishedVersio

Mitogenome diversity of Aedes (Stegomyia) albopictus: Detection of multiple introduction events in Portugal

Aedes albopictus, along with Ae. aegypti, are key arbovirus vectors that have been expanding their geographic range over the last decades. In 2017, Ae. albopictus was detected for the first time at two distinct locations in Portugal. In order to understand how the Ae. albopictus populations recently introduced in Portugal are genetically related and which is their likely route of invasion, we performed an integrative cytochrome C oxidase I gene (COI)- and mitogenome-based phylogeographic analysis of mosquitoes samples collected in Portugal in 2017 and 2018 in the context of the global Ae. albopictus diversity. COI-based analysis (31 partial sequences obtained from 83 mosquitoes) revealed five haplotypes (1 to 5), with haplotype 1 (which is widely distributed in temperate areas worldwide) being detected in both locations. Haplotypes 2 and 3 were exclusively found in Southern region (Algarve), while haplotype 4 and 5 were only detected in the North of Portugal (Penafiel, Oporto region). Subsequent high discriminatory analyses based on Ae. albopictus mitogenome (17 novel sequences) not only confirmed a high degree of genetic variability within and between populations at both geographic locations (compatible with the Ae. albopictus mosquito populations circulating in Europe), but also revealed two mitogenome mutational signatures not previously reported at worldwide level. While our results generally sustain the occurrence of multiple introduction events, fine mitogenome sequence inspection further indicates a possible Ae. albopictus migration within the country, from the Northern introduction locality to the Southern region. In summary, the observed scenario of high Ae. albopictus genetic diversity in Portugal, together with the detection of mosquitoes in successive years since 2017 in Algarve and Penafiel, points that both Ae. albopictus populations seem to be already locally established, as its presence has been reported for three consecutive years, raising the public health awareness for future mosquito-borne diseases outbreaks.Author summary: In 2017, Aedes albopictus was reported for the first time in Portugal at two distinct locations, in the premises of a tire company in Penafiel, in the North, and nearby a golf course in Algarve, a tourism destination in the southernmost country region. The geographical spread of this species is promoted by egg and larvae transport in aquatic trade goods, as tires and aquatic plants, and adult anthropophilic behavior that favors passive land transportation. In Portugal, especially in the Southern region, temperate climate conditions are adequate for adult mosquitoes to survive most of the year. In a way to understand the genetic variability of Ae. albopictus populations introduced in Portugal, we analyzed 31 cytochrome C oxidase I gene (COI) partial sequences and 17 mitogenome sequences, integrating them in the context of the global Ae. albopictus phylogeographic diversity (i.e., 183 COI and 26 mitogenome sequences previously reported at worldwide level). Although COI haplotype 1 predominated, four additional haplotypes (2 to 5) were detected in Portugal. Subsequent in-depth mitogenome analysis revealed considerable genetic diversity, including not only sequences relating to mitogenomes reported mainly from Italy, Japan and China, but also two novel mitogenome mutational signatures. Our study indicates that Ae. albopictus is locally established in Portugal and intra-country dispersal may have already happened, highlighting the challenges for vector surveillance and control programs aiming at restraining arbovirus disease burden in the future.info:eu-repo/semantics/publishedVersio