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1. Epidemiological surveillance of West Nile virus in the world and Brazil

Author

Erica Azevedo Costa, José Joffre Martins Bayeux, Aila Solimar Gonçalves Silva, Guilherme Alves de Queiroz, Beatriz Senra Álvares da Silva Santos, Marcele Neves Rocha, Izabelle Silva Rehfeld, Luisa Feliciano de Souza Franklin, Livia Braga Valle, Maria Isabel Maldonado Coelho Guedes, and Raffaella Bertoni Cavalcanti Teixeira

Subjects
Equine surveillance, West Nile virus, Flavivirus, Awareness, Animal culture, SF1-1100
Abstract

West Nile virus (WNV) is a neurovirulent mosquito-borne Flavivirus that is maintained in nature by a zoonotic transmission cycle between avian hosts and ornithophilic mosquito vectors, mostly from the Culex genus. Until the 1990s, WNV was considered to be an old-world arbovirus, but in 1999, WNV emerged in the United States (US) and spread rapidly, becoming a major threat to public health. WNV adapted to the transmission cycle involving American mosquitoes and birds and reached Central and South America in subsequent years. In 2003, the National West Nile Fever Surveillance System was created in Brazil based on serological screening of animals and sentinel vectors, as recommended by the Pan American Health Organization (PAHO) and the World Health Organization (WHO). Since 2008, serological evidence of WNV infection in Brazilian horses has been reported, and the circulation of WNV has been monitored through the regular serological screening of sentinel horses and reporting of encephalomyelitis cases. Horses are highly susceptible to WNV infection, and outbreaks of neurological disease among horses often precede human cases. In this regard, equine surveillance has been essential in providing early warning to public and animal health authorities in several countries, including Brazil. This demonstrates the need for animal and public health intervention programs to allocate resources to make veterinarians aware of the role they can play in the human surveillance processes by monitoring horses. This review discusses the importance of equine surveillance and the gap that veterinarians can fill on the front line in human surveillance, in Brazil and worldwide, in the context of “One Health”

Published
2020
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2. Construção e avaliação do potencial protetor, em modelo murino, de diferentes construções do Vaccinia Virus Ankara Modificado (MVA) expressando a proteína E de Dengue Virus sorotipos 1, 2 e 4

Author

Sabrynna Brito Oliveira, Flávio Guimarães da Fonseca, Bárbara Resende Quinan, Erna Geessien Kroon, Beatriz Senra Álvares da Silva Santos, Cíntia Lopes de Brito Magalhães, and Marcele Neves Rocha

Subjects
vacina recombinante, Vírus da Dengue, Microbiologia, Vacinas sintéticas, MVA, Vírus Vaccinia
Abstract

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior A dengue é uma das principais arboviroses que afeta o ser humano e um grave problema de saúde pública. Os quatro sorotipos do Dengue virus (DENV1, 2, 3 e 4) são transmitidos através da picada de mosquitos fêmea do gênero Aedes. A vacina disponível atualmente apresenta lacunas imunológicas, o que motiva a continuidade de pesquisas em busca de uma vacina eficaz. O vírus Vaccinia Ankara Modificado (MVA) está entre os vetores virais mais promissores e intensamente estudados mundialmente e é considerado como uma excelente ferramenta para a geração de vacinas. O objetivo dessa pesquisa foi construir MVA recombinantes expressando a proteína E de DENV1, 2 e 4 e avaliar o potencial protetor dessas construções em modelo murino. Genes sintéticos da proteína E de DENV1, 2 e 4 foram desenhados, otimizados, obtidos comercialmente e subclonados no plasmídeo pLW44, dando origem aos plasmídeos de transferência, utilizados na construção dos vírus recombinantes. Culturas de fibroblasto de embrião de galinha (CEF) ou BHK-21 foram infectadas por MVA e transfectadas pelos plasmídeos de transferência pLW44-DENV1, pLW44-DENV2 ou pLW44-DENV4, separadamente, para a construção dos MVAs recombinantes (rMVAs). Os rMVAs gerados tiveram a sua construção confirmada por PCR, sequenciamento e citometria de fluxo. O potencial protetor do rMVA-DENV1 foi testado em camundongos C57/BL6 em esquema de imunização dose-reforço homólogo, seguido de desafio intracranial com o DENV correspondente. Os animais foram imunizados com 107 pfu ou 108 pfu de rMVA-DENV1 e desafiados com DENV1 Mochizucki. Durante o desafio os animais foram monitorados e avaliados quanto ao seu estado geral de saúde pelo protocolo SHIRPA. Com base nos resultados obtidos podemos concluir que as construções dos diferentes vírus recombinantes se deram de maneira eficaz, sobretudo a de rMVA-DENV1, a única testada quanto à capacidade protetora. Os dados gerados nessa pesquisa serão somados aos obtidos anteriormente para DENV3 visando à obtenção de uma vacina tetravalente contra DENV. Dengue is the main arbovirus that affecting humans and a serious public health problem. The four dengue virus serotypes (DENV1, 2, 3 and 4) are transmitted through the bite of female Aedes mosquitoes. A vaccine available worldwide has immunological gaps, which motivates further research for an effective vaccine. The Modified Vaccinia Ankara Virus (MVA) is among the most advanced and intensely studied viral vectors worldwide, and is considered as an excellent tool for vaccine generation. The aim of this work was to construct recombinant MVAs expressing the protein E of DENV1, 2 and 4 and to evaluate the protective potential of these constructs in a murine model. Synthetic genes coding for protein E from DENV1, 2 and 4 were designed, optimized, commercially obtained and subcloned into plasmid pLW44, giving rise to transfer plasmids used in the construction of recombinant viruses. Chicken embryo fibroblast cultures – CEF, or BHK-21 were infected with MVA and transfected with the transfer plasmids pLW44-DENV1, pLW44-DENV2 and pLW44-DENV4 for the construction of recombinant MVAs (rMVA). The generated rMVAs had their correct construction confirmed by PCR, sequencing and flow cytometry. The protective potential of rMVA-DENV1 was tested in C57 / BL6 mice on a homologous dose-boosting immunization schedule, followed by intracranial challenge with the corresponding DENV. Animals were immunized with 107 pfu or 108 pfu of rVA-DENV1 and challenged with DENV1 Mochizuki. During the challenge, they were monitored evaluated through a SHIRPA protocol. Based on the results we can conclude that the construction of the different recombinant viruses was effective, especially rMVA-DENV1, the only one tested for protective capacity. The data generated in this research can be added to those previously obtained for DENV3 in order to obtain a tetravalent vaccine against DENV.

Published
2019

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