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1. Sequencing and Description of the Mitochondrial Genome of Orthopodomyia fascipes (Diptera: Culicidae)

Author

Fábio Silva da Silva, Bruna Laís Sena do Nascimento, Ana Cecília Ribeiro Cruz, Sandro Patroca da Silva, Carine Fortes Aragão, Daniel Damous Dias, Lucas Henrique da Silva e Silva, Lúcia Aline Moura Reis, Hanna Carolina Farias Reis, Liliane Leal das Chagas, José Wilson Rosa Jr., Durval Bertram Rodrigues Vieira, Roberto Carlos Feitosa Brandão, Daniele Barbosa de Almeida Medeiros, and Joaquim Pinto Nunes Neto

Subjects
Culicidae, mitogenome, molecular taxonomy, mosquitoes, phylogenetics, Genetics, QH426-470
Abstract

The genus Orthopodomyia Theobald, 1904 (Diptera: Culicidae) comprises 36 wild mosquito species, with distribution largely restricted to tropical and temperate areas, most of which are not recognized as vectors of epidemiological importance due to the lack of information related to their bionomy and involvement in the cycle transmission of infectious agents. Furthermore, their evolutionary relationships are not completely understood, reflecting the scarcity of genetic information about the genus. Therefore, in this study, we report the first complete description of the mitochondrial genome of a Neotropical species representing the genus, Orthopodomyia fascipes Coquillet, 1906, collected in the Brazilian Amazon region. Using High Throughput Sequencing, we obtained a mitochondrial sequence of 15,598 bp, with an average coverage of 418.5×, comprising 37 functional subunits and a final portion rich in A + T, corresponding to the control region. The phylogenetic analysis, using Maximum Likelihood and Bayesian Inference based on the 13 protein-coding genes, corroborated the monophyly of Culicidae and its two subfamilies, supporting the proximity between the tribes Orthopodomyiini and Mansoniini, partially disagreeing with previous studies based on the use of molecular and morphological markers. The information generated in this study contributes to a better understanding of the taxonomy and evolutionary history of the genus and other groups of Culicidae.

Published
2024
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2. Occurrence of avian reovirus and picobirnavirus in wild birds from an environmental protection area in the Brazilian Amazon

Author

Diego PEREIRA, Lizandra Caroline dos Santos SOUTO, Sylvia de Fátima dos Santos GUERRA, Edvaldo Tavares da PENHA-JÚNIOR, Patrícia dos Santos LOBO, Luana da Silva SOARES, Helder Henrique Costa PINHEIRO, Elaine Hellen Nunes CHAGAS, Ricardo José de Paula Souza e GUIMARÃES, Bruna Alves RAMOS, Liliane Leal das CHAGAS, Maria Nazaré Oliveira FREITAS, Erilene Cristina da Silva FURTADO, Jéssica Cecília Pinheiro RODRIGUES, Alexandre do Rosário CASSEB, Lívia Caricio MARTINS, and Joana D’Arc Pereira MASCARENHAS

Subjects
enteric viruses, epidemiology, RT-PCR, phylogeny, Myrmotherula longipennis, Guira guira, Science (General), Q1-390
Abstract

ABSTRACT Wild birds have great prominence in the transmission of diseases to humans, mainly due to their ease of access to human populations, raising concerns about the potential impact of that proximity on public health. The present study reports ARV and PBV occurrence in wild birds from an environmental protection area in the Amazon biome, in Pará state, Brazil. We obtained 155 fecal specimens from 47 species of wild birds for RNA extraction, ARV and PBV detection utilizing molecular methods, nucleotide sequencing, and phylogenetic analysis. ARV prevalence was 0.6% (1/155), being positive in an individual of Myrmotherula longipennis, and PBV prevalence was 1.29% (2/155), affecting two individuals of Guira guira. The PBV strains were assigned to genogroup I based on phylogenetic analysis, and they shared a strong phylogenetic link with strains isolated from different geographic locations. The ARV strain was more closely related to strains that had previously circulated in the same region. The presence of ARV and PBV in this habitat suggests that infection cycles of these agents occur naturally in the wild ecosystem, potentially triggering transmission events between bird species and humans. This is the first study on ARV detection in wild birds in Brazil and the first report on the occurrence of PBV in wild Guira guira. Additional studies are required to determine the epidemiology, origin, evolution, and emergence of new potentially pathogenic viruses in the Amazon.

Published
2024
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3. INVESTIGAÇÃO SOROLÓGICA PARA AS ARBOVIROSES EM ÁREAS DE EXPLORAÇÃO MINERAL NO ESTADO DO PARÁ

Author

Letícia Françadas Mercês, Camille Ferreira de Oliveira, Franko de Arruda e Silva, Walter Felix Franco Neto, Liliane Leal das Chagas, Éder Barros dos Santos, Milene Silveira Ferreira, and Lívia Caricio Martins

Subjects
Arbovírus Exploração Mineral Sorologia, Infectious and parasitic diseases, RC109-216, Microbiology, QR1-502
Abstract

Introdução: O estado do Pará vem sofrendo grandes transformações ambientais, dentre elas a exploração mineral e o desmatamento, o que aumenta o risco de infecção humana por arbovírus. Assim, objetiva-se demonstrar, através da presença de anticorpos IgM e IgG a possível circulação de arbovírus em residentes e trabalhadores oriundos dos municípios de Parauapebas, no período de 2021 a 2022. Métodos: Foram analisados 532 soros humanos pelo teste de Inibição da Hemaglutinação (IH) para 17 tipos diferentes de arbovírus (Eastern equine encephalitis otma, Western equine encephalitis otma, Mayaro otma, Mucambo otma, Chikungunya otma, Orthobunyavirus tacaiumaense, Orthoflavivirus nilense, Orthoflavivirus flavi cepa selvagem e cepa vacinal (17D), Orthoflavivirus denguei sorotipos 1, 2, 3 e 4, Orthoflavivirus zikaense, Orthoflavivirus louisense, Rocio otma, Orthoflavivirus ilheusense e Orthobunyavirus oropoucheense). A partir dos resultados obtido no teste de IH foram realizada a captura de IgM pelo método de ELISA (MAC-ELISA) em 89 amostras para Mayaro otma, 90 Chikungunya otma, 504 Orthoflavivirus denguei, 504 Orthoflavivirus nilense, 504 Orthoflavivirus louisense, 504 Orthoflavivirus zikaense e 51 Orthobunyavirus oropoucheense. Resultados: Dentre as 532 amostras testadas por IH, 513 (96,43%) apresentaram anticorpos totais. 511 (96,05%) foram positivas para Orthoflavivirus, 101 (18,98%) Alphavirus, 56 (10,53%) Orthobunyavirus. Reação simultânea foi observado em 138 (25,94%) amostras e anticorpos para FAV (vacinal- 17D) em 489 (91,92%). No MAC ELISA, 4 (0,79%) amostras foram positivas para Orthoflavivirus denguei, 1 (0,20%) Orthoflavivirus nilense, 2 (0,40%) Orthoflavivirus louisense, 1 (0,20%) Orthoflavivirus zikaense, 1 (1,96%) Orthobunyavirus oropoucheense. Assim como, 228 (10,15%) apresentaram detecção de anticorpos na zona borderline para uma das espécies testadas e 97 (4,32%) apresentaram reatividade cruzada entre Orthoflavivirus e Orthobunyavirus. Conclusão: Foi possível detectar infecção recente para Orthoflavivirus denguei, Orthoflavivirus zikaense, Orthoflavivirus nilense, Orthobunyavirus oropoucheense e Orthoflavivirus louisense, bem como foi observada a circulação dos demais arbovírus testados na área estudada. Medidas de prevenção das arboviroses e o controle vetorial são fundamentais para evitar surtos e epidemias dessas arboviroses.

Published
2023
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4. First Isolation and Genome Sequence Analysis of West Nile Virus in Mosquitoes in Brazil

Author

Joaquim Pinto Nunes Neto, Lúcia Aline Moura Reis, Maria Nazaré Oliveira Freitas, Bruna Laís Sena do Nascimento, Liliane Leal das Chagas, Hernan Hermes Monteiro da Costa, Jéssica Cecília Pinheiro Rodrigues, Camila Margalho Braga, Eliana Vieira Pinto da Silva, Sandro Patroca Silva, and Lívia Caricio Martins

Subjects
West Nile virus infection, Culicidae, Culex, disease vectors, Medicine
Abstract

West Nile virus is a flavivirus transmitted by mosquitoes, mainly of the genus Culex. In Brazil, serological studies have already indicated the circulation of the virus since 2003, with the first human case detected in 2014. The objective of the present paper is to report the first isolation of WNV in a Culex (Melanoconion) mosquito. Arthropods were collected by protected human attraction and CDC light bait, and taxonomically identified and analyzed by viral isolation, complement fixation and genomic sequencing tests. WNV was isolated from samples of Culex (Melanoconion) mosquitoes, and the sequencing analysis demonstrated that the isolated strain belonged to lineage 1a. The finding of the present study presents the first evidence of the isolation and genome sequencing of WNV in arthropods in Brazil.

Published
2023
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5. Isolation of Flaviviruses and Alphaviruses with Encephalitogenic Potential Diagnosed by Evandro Chagas Institute (Pará, Brazil) in the Period of 1954–2022: Six Decades of Discoveries

Author

Ana Lucia Monteiro Wanzeller, Fabio Silva da Silva, Leonardo Henrique Almeida Hernández, Landerson Junior Leopoldino Barros, Maria Nazaré Oliveira Freitas, Maissa Maia Santos, Ercília de Jesus Gonçalves, Jamilla Augusta Sousa Pantoja, Creuza de Sousa Lima, Maxwell Furtado Lima, Luiz Roberto Oliveira Costa, Liliane Leal das Chagas, Iveraldo Ferreira Silva, Tania Cristina Alves da Silveira da Cunha, Bruna Lais Sena do Nascimento, Helena Baldez Vasconcelos, Elizabeth Salbe Travassos da Rosa, Sueli Guerreiro Rodrigues, Raimunda do Socorro da Silva Azevedo, Lívia Carício Martins, Lívia Medeiros Neves Casseb, Jannifer Oliveira Chiang, Joaquim Pinto Nunes Neto, Ana Cecília Ribeiro Cruz, Valéria Lima Carvalho, Pedro Fernando da Costa Vasconcelos, and Eliana Vieira Pinto da Silva

Subjects
arbovirus, viral isolation, cell culture, neurotropic, public health, Microbiology, QR1-502
Abstract

Viruses with encephalitogenic potential can cause neurological conditions of clinical and epidemiological importance, such as Saint Louis encephalitis virus, Venezuelan equine encephalitis virus, Eastern equine encephalitis virus, Western equine encephalitis virus, Dengue virus, Zika virus, Chikungunya virus, Mayaro virus and West Nile virus. The objective of the present study was to determine the number of arboviruses with neuroinvasive potential isolated in Brazil that corresponds to the collection of viral samples belonging to the Department of Arbovirology and Hemorrhagic Fevers, Evandro Chagas Institute (SAARB/IEC) of the Laboratory Network of National Reference for Arbovirus Diagnosis from 1954 to 2022. In the analyzed period, a total of 1,347 arbovirus samples with encephalitogenic potential were isolated from mice; 5,065 human samples were isolated exclusively by cell culture; and 676 viruses were isolated from mosquitoes. The emergence of new arboviruses may be responsible for diseases still unknown to humans, making the Amazon region a hotspot for infectious diseases due to its fauna and flora species characteristics. The detection of circulating arboviruses with the potential to cause neuroinvasive diseases is constant, which justifies the continuation of active epidemiological surveillance work that offers adequate support to the public health system regarding the virological diagnosis of circulating arboviruses in Brazil.

Published
2023
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6. Arboviruses in Free-Ranging Birds and Hematophagous Arthropods (Diptera, Nematocera) from Forest Remnants and Urbanized Areas of an Environmental Protection Area in the Amazon Biome

Author

Bruna Alves Ramos, Liliane Leal Das Chagas, Franko de Arruda e Silva, Eder Barros dos Santos, Jannifer Oliveira Chiang, Joaquim Pinto Nunes Neto, Durval Bertram Rodrigues Vieira, José Wilson Rosa Junior, Eliana Vieira Pinto da Silva, Maria Nazaré Oliveira Freitas, Maissa Maia Santos, Jamilla Augusta de Sousa Pantoja, Ercília de Jesus Gonçalves, Landeson Junior Leopoldino Barros, Sandro Patroca Silva, Carine Fortes Aragão, Ana Cecilia Ribeiro Cruz, Lívia Medeiros Neves Casseb, Lizandra Caroline dos Santos Souto, Joana D’Arc Pereira Mascarenhas, Erilene Cristina Da Silva Furtado, Raimundo Nelson Souza Da Silva, Alexandre do Rosário Casseb, and Lívia Carício Martins

Subjects
arboviruses, birds, hematophagous arthropods, urbanization, Amazon, Microbiology, QR1-502
Abstract

The rapid and disorderly urbanization in the Amazon has resulted in the insertion of forest fragments into cities, causing the circulation of arboviruses, which can involve hematophagous arthropods and free-ranging birds in the transmission cycles in urban environments. This study aimed to evaluate the circulation of arboviruses in free-ranging birds and hematophagous arthropods captured in an Environmental Protection Area in the Belem metropolitan area, Brazil. Birds were captured using mist nets, and hematophagous arthropods were collected using a human protected attraction technique and light traps. The birds’ sera were subjected to a hemagglutination inhibition test to detect antibodies against 29 arbovirus antigens. Arthropod macerates were inoculated into C6/36 and VERO cell cultures to attempt viral isolation and were tested using indirect immunofluorescence, subsequent genetic sequencing and submitted for phylogenetic analysis. Four bird sera were positive for arbovirus, and one batch of Psorophora ferox was positive for Flavivirus on viral isolation and indirect immunofluorescence. In addition, the Ilheus virus was detected in the sequencing and phylogenetic analysis. The presence of antibodies in sera from free-ranging birds and the isolation of Ilheus virus in Psorophora ferox indicate the circulation of arboviruses in forest remnants in the urban center of Belem.

Published
2022
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7. West Nile Virus in the State of Ceará, Northeast Brazil

Author

Flávia Löwen Levy Chalhoub, Eudson Maia de Queiroz-Júnior, Bruna Holanda Duarte, Marcos Eielson Pinheiro de Sá, Pedro Cerqueira Lima, Ailton Carneiro de Oliveira, Lívia Medeiros Neves Casseb, Liliane Leal das Chagas, Hamilton Antônio de Oliveira Monteiro, Maycon Sebastião Alberto Santos Neves, Cyro Facundo Chaves, Paulo Jean da Silva Moura, Aline Machado Rapello do Nascimento, Rodrigo Giesbrecht Pinheiro, Antonio Roberio Soares Vieira, Francisco Bergson Pinheiro Moura, Luiz Osvaldo Rodrigues da Silva, Kiliana Nogueira Farias da Escóssia, Lindenberg Caranha de Sousa, Izabel Leticia Cavalcante Ramalho, Antônio Williams Lopes da Silva, Leda Maria Simōes Mello, Fábio Felix de Souza, Francisco das Chagas Almeida, Raí dos Santos Rodrigues, Diego do Vale Chagas, Anielly Ferreira-de-Brito, Karina Ribeiro Leite Jardim Cavalcante, Maria Angélica Monteiro de Mello Mares-Guia, Vinícius Martins Guerra Campos, Nieli Rodrigues da Costa Faria, Marcelo Adriano da Cunha e Silva Vieira, Marcos Cesar Lima de Mendonça, Nayara Camila Amorim de Alvarenga Pivisan, Jarier de Oliveira Moreno, Maria Aldessandra Diniz Vieira, Ricristhi Gonçalves de Aguiar Gomes, Fernanda Montenegro de Carvalho Araújo, Pedro Henrique de Oliveira Passos, Daniel Garkauskas Ramos, Alessandro Pecego Martins Romano, Lívia Carício Martins, Ricardo Lourenço-de-Oliveira, Ana Maria Bispo de Filippis, and Alex Pauvolid-Corrêa

Subjects
West Nile virus, Brazil, equids, Boa Viagem, Ceará, PRNT, Biology (General), QH301-705.5
Abstract

In June 2019, a horse with neurological disorder was diagnosed with West Nile virus (WNV) in Boa Viagem, a municipality in the state of Ceará, northeast Brazil. A multi-institutional task force coordinated by the Brazilian Ministry of Health was deployed to the area for case investigation. A total of 513 biological samples from 78 humans, 157 domestic animals and 278 free-ranging wild birds, as well as 853 adult mosquitoes of 22 species were tested for WNV by highly specific serological and/or molecular tests. No active circulation of WNV was detected in vertebrates or mosquitoes by molecular methods. Previous exposure to WNV was confirmed by seroconversion in domestic birds and by the detection of specific neutralizing antibodies in 44% (11/25) of equids, 20.9% (14/67) of domestic birds, 4.7% (13/278) of free-ranging wild birds, 2.6% (2/78) of humans, and 1.5% (1/65) of small ruminants. Results indicate that not only equines but also humans and different species of domestic animals and wild birds were locally exposed to WNV. The detection of neutralizing antibodies for WNV in free-ranging individuals of abundant passerine species suggests that birds commonly found in the region may have been involved as amplifying hosts in local transmission cycles of WNV.

Published
2021
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8. Clinical and serological tests for arboviruses in free-living domestic pigeons (Columba livia)

Author

Bruna Alves Ramos, Jannifer Oliveira Chiang, Lívia Carício Martins, Liliane Leal das Chagas, Franko de Arruda e Silva, Milene Silveira Ferreira, Maria Nazaré Oliveira Freitas, Bianca Nascimento de Alcantara, Sandro Patroca da Silva, Stefânia Araújo Miranda, Barbara Alves Sepulvreda, Layna Thayssa Guimarães Corrêa, Andréa Maria Góes Negrão, Pedro Fernando da Costa Vasconcelos, and Alexandre do Rosário Casseb

Subjects
arbovirus, domestic pigeons, clinical, serological tests, Microbiology, QR1-502, Infectious and parasitic diseases, RC109-216
Abstract

BACKGROUND In this study, we evaluated the role of free-living domestic pigeons (Columba livia) as a reservoir of arboviruses in the city of Belém, state of Pará, Brazil. We investigated the presence of antibodies against the most prevalent arboviruses. OBJECTIVES This study was aimed at evaluating some clinical and physical parameters of domestic pigeons, including the presence of antibodies to Amazon-endemic arboviruses. METHODS Eighty-five healthy pigeons were captured in Mangal das Garças Park, in Belém, and were bled. Upon capture, the birds were subjected to a clinical examination in search of alterations that could indicate the presence of arboviruses. Blood samples were converted to serum and tested using the haemagglutination inhibition (HI) technique with a panel of 19 antigens of arboviruses circulating in the Amazon. The confirmation assay for the positive reactions to the viral species tested by HI was a neutralisation test in new-born Swiss albino mice (Mus musculus) [mouse neutralisation test (MNT)]. FINDINGS A total of 10 (11.8%) serum samples tested positive for antiflavivirus antibodies by HI. All the samples positive for the HI test were subjected to MNT for detection of viruses and yielded negative results (logarithmic neutralisation index < 1.7). MAIN CONCLUSION The results represent the first serological detection of antiarbovirus antibodies in domestic pigeons as potential hosts of arboviruses in Brazil. The detection of haemagglutination-inhibiting antibodies against genus Flavivirus indicated that there was recent contact between the analysed domestic pigeons and these arboviruses. Further studies are needed to evaluate the role of free-living pigeons in the maintenance cycle and spread of arboviruses in the Amazon.

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9. Isolation of Flaviviruses and Alphaviruses with Encephalitogenic Potential Diagnosed by Evandro Chagas Institute in the Period of 1954–2022: Six Decades of Discoveries

Author

Ana Lucia Monteiro Wanzeller, Landeson Junior Leopoldino Barros, Maria Nazaré Oliveira Freitas, Maissa Maia Santos, Ercília de Jesus Gonçalves, Jamilla Pantoja, Creuza de Sousa Lima Lima, Maxuell Furtado Lima, Luiz Roberto Oliveira Costa, Liliane Leal das Chagas, Iveraldo Ferreira Silva, Tania Cristina Alves da Silveira da Cunha, Fabio Silva da Silva, Bruna Laís Sena do Nascimento, Leonardo Henrique Almeida Hernández, Raimunda do Socorro da Silva Azevedo, Lívia Carício Martins, Jannifer Oliveira Chiang, Joaquim Pinto Nunes Neto, Ana Cecília Ribeiro Cruz, Livia Medeiros Neves Casseb, Elizabeth Salbe Travassos da Rosa, Sueli Guerreiro Rodrigues, Pedro Fernando da Costa Vasconcelos, Helena Baldez Vasconcelos, Valéria Lima Carvalho, and Eliana Vieira Pinto da Silva

Subjects
virology
Abstract

Viruses with encephalitogenic potential can cause neurological conditions of clinical and epidemiological importance, such as Saint Louis encephalitis virus, Venezuelan equine encephalitis virus, Eastern equine encephalitis virus, Western equine encephalitis virus, dengue virus, Zika virus, Chikungunya virus, Mayaro virus and West Nile virus. The objective of the present study was to determine the number of arboviruses with neuroinvasive potential isolated in Brazil that corresponds to the collection of viral samples belonging to the Section of Arbovirology and Hemorrhagic Fevers (SAARB/IEC) of the Laboratory Network of National Reference for Arbovirus Diagnosis from 1954 to 2022. A total of 1,090 arbovirus samples with encephalitogenic potential were isolated from mice. From a total of 7,054 samples intended for cell culture. The isolates from mosquitoes totaled 659 viruses. The emergence of new arboviruses may be responsible for diseases still unknown to humans, making the Amazon region a hotspot for infectious diseases due to its fauna and flora species characteristics. The detection of circulating arboviruses with the potential to cause neuroinvasive diseases is constant, which justifies the continuation of active epidemiological surveillance work that offers adequate support to the public health system regarding the virological diagnosis of circulating arboviruses in Brazil.

Published
2023

10. West Nile Virus in the State of Ceará, Northeast Brazil

Author

Maria Aldessandra Diniz Vieira, Ailton Carneiro de Oliveira, Paulo Jean da Silva Moura, Fernanda Montenegro de Carvalho Araújo, Lívia Carício Martins, Fábio Felix de Souza, Karina Ribeiro Leite Jardim Cavalcante, Antônio Williams Lopes da Silva, Kiliana Nogueira Farias da Escóssia, Eudson Maia de Queiroz-Júnior, Alessandro Pecego Martins Romano, Cyro Facundo Chaves, Maria Angélica Mares-Guia, Jarier de Oliveira Moreno, Nieli Rodrigues da Costa Faria, Vinícius Martins Guerra Campos, Pedro Henrique de Oliveira Passos, Anielly Ferreira-de-Brito, Ricristhi Gonçalves de Aguiar Gomes, Flávia Löwen Levy Chalhoub, Ricardo Lourenço-de-Oliveira, Aline Machado Rapello do Nascimento, Pedro Cerqueira Lima, Livia Medeiros Neves Casseb, Alex Pauvolid-Corrêa, Francisco Bergson Pinheiro Moura, Rodrigo Giesbrecht Pinheiro, Marcelo Adriano da Cunha e Silva Vieira, Marcos Eielson Pinheiro de Sá, Daniel Garkauskas Ramos, Raí dos Santos Rodrigues, Nayara Camila Amorim de Alvarenga Pivisan, Maycon Sebastião Alberto Santos Neves, Marcos Cesar Lima de Mendonça, Liliane Leal das Chagas, Diego do Vale Chagas, Bruna Holanda Duarte, Izabel Letícia Cavalcante Ramalho, Francisco das Chagas Almeida, Lindenberg Caranha de Sousa, Ana Maria Bispo de Filippis, Hamilton Antônio de Oliveira Monteiro, Leda Maria Simōes Mello, Luiz Osvaldo Rodrigues da Silva, and Antonio Roberio Soares Vieira

Subjects
Microbiology (medical), West Nile virus, QH301-705.5, equids, Zoology, Northeast brazil, medicine.disease_cause, Microbiology, V?rus do Nilo Ocidental / patogenicidade, Article, Serology, Boa Viagem, Virology, biology.animal, medicine, Seroconversion, Biology (General), Febre do Nilo Ocidental / veterin?ria, biology, Task force, Transmission (medicine), PRNT, virus diseases, Ceará, Passerine, Exposi??o Ambiental / an?lise, Cavalos / anatomia & histologia, Febre do Nilo Ocidental / transmiss?o, Christian ministry, Cavalos / virologia, Brazil
Abstract

Funda??o de Amparo ? Pesquisa do Estado do Rio de Janeiro (Faperj) under the grant no. E-26/202.930/2016, European Union?s Horizon 2020 grant agreement ZIKACTION no. 734857 and Coordena??o de Vigil?ncia em Sa?de e Laborat?rios de Refer?ncia da Funda??o Oswaldo Cruz/Minist?rio de Sa?de. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Ag?ncia de Defesa Agropecu?ria do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Minist?rio da Agricultura Pecu?ria e Abastecimento. Departamento de Servi?os T?cnicos. Secretaria de Defesa Agropecu?ria. Bras?lia, DF, Brazil. Funda??o BioBrasil. Ituber?, BA, Brazil. Minist?rio do Meio Ambiente. Instituto Chico Mendes de Conserva??o da Biodiversidade. Centro Nacional de Pesquisa para Conserva??o das Aves Silvestres. Cabedelo, PB, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswaldo Cruz. Laborat?rio de Mosquitos Transmissores de Hematozo?rios. Rio de Janeiro, RJ, Brazil. Autonomous Veterinarian. Boa Viagem, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Laborat?rio Central do Estado do Cear? . Fortaleza, CE, Brazil. Ag?ncia de Defesa Agropecu?ria do Estado do Cear?. Fortaleza, CE, Brazil. Laborat?rio Central do Estado do Cear? . Fortaleza, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Funda??o Oswaldo Cruz. Laborat?rio de Mosquitos Transmissores de Hematozo?rios. Rio de Janeiro, RJ, Brazil. Coordena??o Geral de Laborat?rios de Sa?de P?blica. Bras?lia, DF, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil / Secretaria de Estado da Sa?de do Piau?. Coordena??o de Epidemiologia. Teresina, PI, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Ag?ncia de Defesa Agropecu?ria do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria Municipal de Sa?de de Boa Viagem. Boa Viagem, CE, Brazil. Secretaria Estadual de Sa?de do Estado do Cear?. Fortaleza, CE, Brazil. Laborat?rio Central do Estado do Cear?. Fortaleza, CE, Brazil. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil. Secretaria de Vigil?ncia em Sa?de. Coordena??o-Geral de Vigil?ncia das Arboviroses. Departamento de Imuniza??o e Doen?as Transmiss?veis. Bras?lia, DF, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o Oswlado Cruz. Laborat?rio de Mosquitos Transmissores de Hematozo?rios. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Funda??o Oswaldo Cruz. Laborat?rio de Flaviv?rus. Rio de Janeiro, RJ, Brazil / Texas A&M University. College Station. Department of Veterinary Integrative Biosciences. TX, USA. Abstract: In June 2019, a horse with neurological disorder was diagnosed with West Nile virus (WNV) in Boa Viagem, a municipality in the state of Cear?, northeast Brazil. A multi-institutional task force coordinated by the Brazilian Ministry of Health was deployed to the area for case investigation. A total of 513 biological samples from 78 humans, 157 domestic animals and 278 free-ranging wild birds, as well as 853 adult mosquitoes of 22 species were tested for WNV by highly specific serological and/or molecular tests. No active circulation of WNV was detected in vertebrates or mosquitoes by molecular methods. Previous exposure to WNV was confirmed by seroconversion in domestic birds and by the detection of specific neutralizing antibodies in 44% (11/25) of equids, 20.9% (14/67) of domestic birds, 4.7% (13/278) of free-ranging wild birds, 2.6% (2/78) of humans, and 1.5% (1/65) of small ruminants. Results indicate that not only equines but also humans and different species of domestic animals and wild birds were locally exposed to WNV. The detection of neutralizing antibodies for WNV in free-ranging individuals of abundant passerine species suggests that birds commonly found in the region may have been involved as amplifying hosts in local transmission cycles of WNV.

Published
2021
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11. Reporter Virus Neutralization Test Evaluation for Dengue and Zika Virus Diagnosis in Flavivirus Endemic Area

Author

Eliana Vieira Pinto da Silva, Liliane Leal das Chagas, Raimunda do Socorro da Silva Azevedo, Rommel Mario Rodrigues Burbano, Pei Yong Shi, Bruno Tardelli Diniz Nunes, Jannifer Oliveira Chiang, Daniele Barbosa de Almeida Medeiros, Tais R Silva, Jannyce G. C Nunes, Lívia Carício Martins, Maria Helena Rodrigues de Mendonça, Livia Medeiros Neves Casseb, Daniele Freitas Henriques, Pedro Fernando da Costa Vasconcelos, Adriana Freitas Moraes, Chao Shan, and Franco A. e Silva

Subjects
0301 basic medicine, Microbiology (medical), diagnosis, viruses, Secondary infection, 030231 tropical medicine, Virus Neutralization, Article, reporter virus, Serology, Dengue fever, Zika virus, Dengue, 03 medical and health sciences, V?rus Rep?rter, 0302 clinical medicine, flavivirus, Immunology and Allergy, Medicine, Seroconversion, Molecular Biology, neutralization test, ZIKV, General Immunology and Microbiology, biology, business.industry, V?rus da Dengue / patogenicidade, Outbreak, biology.organism_classification, medicine.disease, Virology, Zika virus / patogenicidade, Flavivirus, 030104 developmental biology, Infectious Diseases, Testes de Neutraliza??o / m?todos, Flavivirus / isolamento & purifica??o, business
Abstract

Ministry of Health of Brazil and by the grants from CNPq (process 303999/2016-0 and 440405/2016-5) and CAPES (Zika fast track project). Pan American Health Organization SCON2016-01353, NIH grants AI142759, AI127744, and AI136126, and awards from the Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, Gilson Longenbaugh Foundation, and Summerfield Robert Foundation. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / State University of Par?. Post Graduation Program in Parasitary Biology in the Amazon. Bel?m, PA, Brazil. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Par? State University. Science and Health Institute. Bel?m, PA, Brazil. Federal University of Par?. Biological Sciences Institute. Bel?m, PA, Brazil. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA . University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / State University of Par?. Post Graduation Program in Parasitary Biology in the Amazon. Bel?m, PA, Brazil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Reporter virus neutralization test (RVNT) has been used as an alternative to the more laborious and time-demanding conventional PRNT assay for both DENV and ZIKV. However, few studies have investigated how these techniques would perform in epidemic areas with the circulation of multiple flavivirus. Here, we evaluate the performance of ZIKV and DENV Rluc RVNT and ZIKV mCh RVNT assays in comparison to the conventional PRNT assay against patient sera collected before and during ZIKV outbreak in Brazil. These samples were categorized into groups based on (1) acute and convalescent samples according to the time of disease, and (2) laboratorial diagnostic results (DENV and ZIKV RT-PCR and IgM-capture ELISA). Our results showed that DENV Rluc assay presented 100% and 78.3% sensitivity and specificity, respectively, with 93.3% accuracy, a similar performance to the traditional PRNT. ZIKV RVNT90, on the other hand, showed much better ZIKV antibody detection performance (around nine-fold higher) when compared to PRNT, with 88% clinical sensitivity. Specificity values were on average 76.8%. Even with these results, however, ZIKV RVNT90 alone was not able to reach a final diagnostic conclusion for secondary infection in human samples due to flavivirus cross reaction. As such, in regions where the flavivirus differential diagnosis represents a challenge, we suggest the establishment of a RVNT panel including other flaviviruses circulating in the region, associated with the other serological techniques such as IgM ELISA and the investigation of seroconversion, in order to help define an accurate diagnostic conclusion using serology.

Published
2021

12. Clinical and serological tests for arboviruses in free-living domestic pigeons (Columba livia)

Author

Pedro Fernando da Costa Vasconcelos, Liliane Leal das Chagas, Milene Silveira Ferreira, Jannifer Oliveira Chiang, Alexandre do Rosário Casseb, Bianca Nascimento de Alcantara, Layna Thayssa Guimarães Corrêa, Barbara Alves Sepulvreda, Maria Nazaré Oliveira Freitas, Stefânia Araújo Miranda, Franko de Arruda e Silva, Lívia Carício Martins, Bruna Alves Ramos, Sandro Patroca da Silva, and Andréa Maria Góes Negrão

Subjects
Male, serological tests, 0301 basic medicine, Microbiology (medical), Veterinary medicine, lcsh:Arctic medicine. Tropical medicine, Columbidae / parasitologia, lcsh:RC955-962, domestic pigeons, viruses, Testes de Inibi??o da Hemaglutina??o / m?todos, 030106 microbiology, 030231 tropical medicine, lcsh:QR1-502, Arbovirus Infections, Disease Vectors, Parque Mangal das Gar?as (PA), Antibodies, Viral, lcsh:Microbiology, clinical, Neutralization, Serology, Haemagglutination inhibition, 03 medical and health sciences, 0302 clinical medicine, Antigen, Genus Flavivirus, parasitic diseases, Animals, Columbidae, Arbovirus, Reservat?rios de Doen?as, biology, Bird Diseases, virus diseases, Bel?m (PA), Articles, Hemagglutination Inhibition Tests, Serum samples, arbovirus, biology.protein, Female, Antibody, Arboviruses, Brazil
Abstract

To Fapespa, for granting a Master?s scholarship, to PNMG, for capture of the birds, to the Microbiology and Immunology Laboratory of the UFRA, for collection of the samples, and to the SAARB/IEC, for their support in the development of serological tests. Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Parque Natural?stico Mangal das Gar?as. Setor Veterin?rio. Bel?m, PA, Brasil. Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Bel?m, PA, Brasil. Universidade Federal Rural da Amaz?nia. Instituto da Sa?de e Produ??o Animal. Laborat?rio de Biologia Molecular. Bel?m, PA, Brasil. Universidade Federal Rural da Amaz?nia. Instituto da Sa?de e Produ??o Animal. Laborat?rio de Biologia Molecular. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade Federal Rural da Amaz?nia. Instituto da Sa?de e Produ??o Animal. Laborat?rio de Biologia Molecular. Bel?m, PA, Brasil. BACKGROUND: In this study, we evaluated the role of free-living domestic pigeons (Columba livia) as a reservoir of arboviruses in the city of Bel?m, state of Par?, Brazil. We investigated the presence of antibodies against the most prevalent arboviruses. OBJECTIVES: This study was aimed at evaluating some clinical and physical parameters of domestic pigeons, including the presence of antibodies to Amazon-endemic arboviruses. METHODS: Eighty-five healthy pigeons were captured in Mangal das Gar?as Park, in Bel?m, and were bled. Upon capture, the birds were subjected to a clinical examination in search of alterations that could indicate the presence of arboviruses. Blood samples were converted to serum and tested using the haemagglutination inhibition (HI) technique with a panel of 19 antigens of arboviruses circulating in the Amazon. The confirmation assay for the positive reactions to the viral species tested by HI was a neutralisation test in new-born Swiss albino mice (Mus musculus) [mouse neutralisation test (MNT)]. FINDINGS: A total of 10 (11.8%) serum samples tested positive for antiflavivirus antibodies by HI. All the samples positive for the HI test were subjected to MNT for detection of viruses and yielded negative results (logarithmic neutralisation index < 1.7). MAIN CONCLUSION: The results represent the first serological detection of antiarbovirus antibodies in domestic pigeons as potential hosts of arboviruses in Brazil. The detection of haemagglutination-inhibiting antibodies against genus Flavivirus indicated that there was recent contact between the analysed domestic pigeons and these arboviruses. Further studies are needed to evaluate the role of free-living pigeons in the maintenance cycle and spread of arboviruses in the Amazon.

Published
2017

13. Investigation about the Occurrence of Transmission Cycles of Arbovirus in the Tropical Forest, Amazon Region

Author

Lívia Carício Martins, Thito Bezerra Paz, Leonardo Almeida Hernández, Eliana Vieira Pinto da Silva, Landeson L. Barros, Liliane Leal das Chagas, Joaquim P. Nunes-Neto, Milene Silveira Ferreira, Samir Mansour Moraes Casseb, José Wilson Rosa-Júnior, Jannifer Oliveira Chiang, Pedro Arthur da Silva Araújo, Pedro Fernando da Costa Vasconcelos, B. L. S Nascimento, Maria Nazaré Oliveira Freitas, Sandro Patroca da Silva, Raimunda do Socorro da Silva Azevedo, and Franko de Arruda e Silva

Subjects
0106 biological sciences, 0301 basic medicine, Ciclos de Transmiss?o, Hemagglutination, Arbov?rus / crescimento & desenvolvimento, viruses, Biodiversity, lcsh:QR1-502, 01 natural sciences, Genome, lcsh:Microbiology, law.invention, law, biodiversity, Mammals, education.field_of_study, Amazon rainforest, Biodiversidade, Caixuanã, Vetores de Doen?as, Infectious Diseases, Transmission (mechanics), Floresta Nacional de Caxiuan? (PA), Brazil, Rainforest, Population, Zoology, Genome, Viral, Mosquito Vectors, Biology, Arbovirus Infections, Arbovirus, Article, Birds, 03 medical and health sciences, Virology, medicine, Animals, education, Amazon, Hemagglutination assay, Reservat?rios de Doen?as, Genoma Viral / gen?tica, Hemagglutination Inhibition Tests, medicine.disease, 010602 entomology, 030104 developmental biology, Culicidae, arbovirus, Arboviruses, Pará
Abstract

Because of its ecological characteristics, the Caxiuan&atilde, National Forest (FLONA) is a potential area of arbovirus circulation. The present study aimed to investigate the occurrence of arbovirus transmission cycles at FLONA de Caxiuan&atilde, Five field trips were performed to capture mosquitoes and sylvatic vertebrates. For these vertebrates, we attempted viral isolation by cell monolayer inoculation from blood, and hemagglutination inhibition and further seroneutralization assays from sera. For mosquitoes, we performed tests of viral genome detection. A total of 338 vertebrates were captured, and the greatest representative was birds (251/74.26%). A total of 16,725 mosquitoes were captured, distributed among 56 species. There were no viruses isolated by newborn mouse inoculation. Among birds, antibodies against Ilheus virus were the most prevalent. Catu virus, Caraparu virus, and Mucambo virus were the most prevalent among mammals and reptiles. Fragments of Mucambo virus, Ilheus virus, Bussuquara virus, and Rocio virus genome were detected in a pool of mosquito samples. These results of the study suggest the occurrence of arbovirus transmission cycles in the FLONA of Caxiuan&atilde, The proximity of human populations with elements, involved in transmission cycles, makes surveillance necessary in this population to avoid dispersion of arboviruses to na&iuml, ve locations.

Published
2019
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