Monitoramento das Produções

Title: Production of yellow fever virus in microcarrier-based Vero cell cultures
Authors: Souza, Marta Cristina O.; Freire, Marcos S.; Schulze, Erica A.; Gaspar, Luciane P.; Castilho, Leda R.

Title: As Políticas de Educação Profissional em Saúde no Contexto da Dengue: a perspectiva dos agentes de combate às endemias
Authors: Evangelista, Janete Gonçalves
Abstract: A partir de um caráter multiprofissional e interdisciplinar, constitutivo da problemática da dengue, este trabalho tem por objetivo principal analisar os aspectos que constituem a qualificação profissional dos Agentes de Combate às Endemias (ACE), bem como suas vivências relacionadas à sua formação e práticas de trabalho. A partir de uma abordagem qualitativa, o percurso metodológico estruturou-se conforme as seguintes etapas: (1) Análise documental das diretrizes e programas de formação profissional voltados para os ACE no que se refere ao tema da dengue; (2)
Análise dos processos de construção de identidades dos ACE relacionados a sua formação e práticas de trabalho, a partir de grupos focais. Considera-se que ações e políticas mais integradas e intersetoriais podem auxiliar nas práticas de formação e de trabalho do ACE, auxiliando-o a se reconhecer como pertencente a uma categoria profissional, com espaço específico para sua organização e mobilização político-institucional. Verificou-se, ainda, que uma minoria dos ACE que participaram na pesquisa recebeu formação precípua. Todos desconheciam o protocolo de execução de suas atividades e, definiam por si mesmos os processos de trabalho em seu cotidiano. Os agentes apresentaram dificuldade de se identificarem como
pertencentes à categoria “trabalhadores da saúde”. Essas condições dificultaram o seu reconhecimento por parte da sociedade e pelo próprio setor da saúde, tornando-os “invisíveis” ou “não identificados”. Assim, as identidades possíveis construídas pelos ACE são aquelas alicerçadas no vínculo afetivo e social com os moradores e com o seu cotidiano de trabalho. Conclui-se que a institucionalização de uma política de educação profissional sólida e direcionada aos ACE, bem como o fortalecimento
das identidades profissionais desses agentes, podem contribuir para um avanço significativo na prevenção e no controle de vetores e da dengue.

Title: Production of yellow fever 17DD vaccine virus in primary culture of chicken embryo fibroblasts: yields, thermo and genetic stability, attenuation and immunogenicity
Authors: Freire, Marcos S.; Mann, George F.; Marchevsky, Renato S.; Yamamura, Anna M.Y.; Almeida, Luiz F. C.; Jabor, Alfredo V.; Malachias, José M. N.; Coutinho, Evandro S. F.; Galler, Ricardo

by Armel N. Tedjou, Basile Kamgang, Aurélie P. Yougang, Flobert Njiokou, Charles S. Wondji
Introduction Arboviral diseases including dengue are increasingly spreading in the tropical/subtropical world including Africa. Updated knowledge on the distribution and abundance of the major vectors Aedes aegypti and Aedes albopictus constitutes crucial surveillance action to prepare African countries such as Cameroon for potential arbovirus outbreaks. Here, we present a nationwide survey in Cameroon to assess the current geographical distribution and prevalence of both vectors including a genetic diversity profiling of Ae. albopictus (invasive species) using mitochondrial DNA. Methods Immature stages of Aedes were collected between March and August 2017 in 29 localities across Cameroon following north-south and east-west transects. Larvae and pupae were collected from several containers in each location, reared to adult and morphologically identified. Genetic diversity of Ae. albopictus from 16 locations were analysed using Cytochrome Oxidase I gene (COI). Results In total, 30,381 immature stages of Aedes with an average of 646.40±414.21 per location were identified across the country comprising 69.3% of Ae. albopictus and 30.7% of Ae. aegypti. Analysis revealed that Ae. aegypti is still distributed nation widely whereas Ae. albopictus is limited to the southern part, around 6°4’N. However, Ae. albopictus is the most prevalent species in all southern locations where both species are sympatric except in Douala where Ae. aegypti is predominant. This suggests that factors such as climate, vegetation, and building density impact the distribution of both species in Cameroon. Mitochondrial DNA analysis revealed a low genetic diversity in Ae. albopictus populations with a major common haplotype resulting in low haplotype diversity ranging from 0.13 to 0.65 and 0.35 for the total sample. Similarly, low nucleotide diversity was also reported varying from 0.0000 to 0.0017 with an overall index of 0.0008. This low genetic polymorphism is consistent with the recent introduction of Ae. albopictus in Cameroon. Conclusion This updated distribution of arbovirus vectors across Cameroon will help in planning vector control programme against possible outbreak of arbovirus related diseases in the country.

Title: Aedes Aegypti, vetor de epidemias anunciadas
Authors: Fundação Oswaldo Cruz. Gerência Regional de Brasília
Abstract: Apresenta publicações de textos extraídos das apresentações e reflexões elaboradas pelos autores convidados dos seminários As Relações da Saúde Pública com a Imprensa (RSPI), que, em 2017, teve sua primeira edição internacional. Debate sobre as possibilidades e as responsabilidades de todos os atores envolvidos na difícil — porém necessária — tarefa de fazer saúde em todas as suas dimensões, sejam elas práticas, sejam elas simbólicas. Neste número inaugural, são apresentados os olhares de um amplo leque de atores, que ajudam a compreender que papel a mídia exerce e pode exercer em suas coberturas sobre os impactos causados pela inescapável presença do Aedes aegypti em nossas vidas. Autores brasileiros e de outros países narram experiências e fazem refl exões sobre de que forma a população deve ser
mobilizada a agir em prol de sua própria saúde sem abrir mão da responsabilidade e presença do Estado na edificação de políticas e ações que garantam as condições adequadas para a saúde de todos. A trajetória dos seminários RSPI tem seu ponto de partida em 2008, quando o Distrito Federal viveu meses nebulosos de informações e desinformações sobre uma possível epidemia de febre amarela. Naquela época, em que as
mídias sociais ainda não pautavam o cotidiano das pessoas com a mesma intensidade com que o fazem hoje, a mídia tradicional exercia a centralidade na construção dos sentidos sobre saúde e sobre doença. Fato é que uma sensação muito próxima ao pânico se estabeleceu na capital do Brasil, fazendo com que pessoas corressem aos postos de saúde nem sempre para se vacinar, mas, em muitos casos, se re-vacinar.
Neste sentido, a Fiocruz Brasília, por meio de sua Assessoria de Comunicação, se viu instada a agir no sentido de estabelecer pontes entre agentes públicos estratégicos e promover reflexões sobre como imprensa e saúde pública podem interagir para assegurar o bem da sociedade. Assim, no auditório da Organização Pan-Americana da Saúde (Opas), especialistas da área da saúde, jornalistas e analistas da imprensa se aprofundaram, ao longo de apenas uma manhã, nas questões mais marcantes da relação saúde pública e imprensa.

Title: EAD e febre amarela: reflexões acerca de uma proposta de curso para os trabalhadores de Bio-Manguinhos
Authors: Assed, Marcela Luiza Soares
Abstract: Febre Amarela (FA) é uma doença infecciosa causada por um vírus e transmitida por mosquitos e pode, segundo a Organização Mundial de Saúde (OMS), ser categorizada em selvática, intermédia e urbana, este último onde o vírus é introduzido em zonas densamente povoadas. O Instituto Tecnológico em Imunobiológicos (Bio-Manguinhos/Fiocruz), é o maior produtor mundial da vacina contra a doença, principal fornecedor do Ministério da Saúde (MS) por meio do Programa Nacional de Imunizações (PNI). Há relatos de surto urbano da doença desde 1730, mas as recentes epidemias na África e Brasil preocuparam os epidemiologistas e evidenciaram a necessidade da instituição de apresentar aos seus trabalhadores, informações e esclarecimentos no tema. Entendeu-se tático não apenas informar, mas desenvolver uma experiência piloto, utilizando EaD, enquanto ferramenta que se propicia, de modo autônomo, reflexão e construção coletiva no conjunto de iniciativas já implementadas na organização de acordo com as políticas públicas vigentes. Por meio da investigação bibliográfica, referenciada no materialismo histórico dialético, apresenta-se ao fim da dissertação as análises necessárias para proposição de um curso a distância, de domínio público no tema definido. Para alicerce pedagógico, além da consulta a documentos normativos sobre EaD foi realizado a análise dos Referenciais de Qualidade para a EaD em busca da garantia das normas de qualidade no processo de ensino/aprendizagem e o caráter emancipador da educação politécnica.

by Priscila M. S. Castanha, Wayner V. Souza, Cynthia Braga, Thalia V. B. De Araújo, Ricardo A. A. Ximenes, Maria de Fátima P. M. Albuquerque, Ulisses R. Montarroyos, Demócrito B. Miranda-Filho, Marli T. Cordeiro, Rafael Dhalia, Ernesto T. A. Marques Jr., Laura C. Rodrigues, Celina M. T. Martelli, Microcephaly Epidemic Research Group

Laboratory confirmation of Zika virus (ZIKV) infection during pregnancy is challenging due to cross-reactivity with dengue virus (DENV) and limited knowledge about the kinetics of anti-Zika antibody responses during pregnancy. We described ZIKV and DENV serological markers and the maternal-fetal transfer of antibodies among mothers and neonates after the ZIKV microcephaly outbreak in Northeast Brazil (2016). We included 89 microcephaly cases and 173 neonate controls at time of birth and their mothers. Microcephaly cases were defined as newborns with a particular head circumference (2 SD below the mean). Two controls without microcephaly were matched by the expected date of delivery and area of residence. We tested maternal serum for recent (ZIKV genome, IgM and IgG3 anti-NS1) and previous (ZIKV and DENV neutralizing antibodies [NAbs]) markers of infection. Multiple markers of recent or previous ZIKV and DENV infection in mothers were analyzed using principal component analysis (PCA). At delivery, 5.6% of microcephaly case mothers and 1.7% of control mothers were positive for ZIKV IgM. Positivity for ZIKV IgG3 anti-NS1 was 8.0% for case mothers and 3.5% for control mothers. ZIKV NAbs was slightly higher among mothers of cases (69.6%) than that of mothers of controls (57.2%; p = 0.054). DENV exposure was detected in 85.8% of all mothers. PCA discriminated two distinct components related to recent or previous ZIKV infection and DENV exposure. ZIKV NAbs were higher in newborns than in their corresponding mothers (p

Title: Detection of TH1/TH2 cytokine signatures in yellow fever 17DD first-time vaccinees through ELISpot assay
Authors: Santos, A. P.; Matos, D. C. S.; Bertho, A. L.; Mendonça, S. C. F.; Marcovistz, R.

Title: Comparison of different uses of adult traps and ovitraps for assessing dengue vector infestation in endemic areas
Authors: Oliveira, Ricardo Lourenço de; Lima, José Bento P.; Peres, Roberto; Alves, Fernando da Costa; Eiras, Álvaro E.; Codeço, Cláudia Torres

Title: Zika Virus Epidemic in Brazil. II. Post-Mortem Analyses of Neonates with Microcephaly, Stillbirths, and Miscarriage
Authors: Azevedo, Raimunda S. S.; Araujo, Marialva T.; Oliveira, Consuelo S.; Filho, Arnaldo Jorge Martins; Nunes, Bruno T. D.; Henriques, Daniele F.; Silva, Eliana V. P.; Carvalho, Valéria L.; Chiang, Jannifer O.; Martins, Lívia C.; Vasconcelos, Barbara C. B.; Sousa, Jorge R.; Araujo, Fernanda Montenegro C.; Ribeiro, Erlane M.; Castro, Andrezza R. P.; de Queiroz, Maria G. L.; Verotti, Mariana P.; Nunes, Márcio R. T.; Cruz, Ana C. R.; Rodrigues, Sueli G.; Shi, Pei-Yong; Quaresma, Juarez A. S.; Tesh, Robert B.; Vasconcelos, Pedro F. C.
Description: We are grateful for the staff of the Department of Pathology at the IEC, especially Valter
Miranda Campos and Maria de Lourdes Gomes Lima, personal of the Department of Arbovirology and
Hemorrhagic Fevers at the IEC, and Adlai Raimundo Sousa for their technical support, as well as staff of
the Public Health Laboratories of Ceará, Espírito Santo, Pernambuco, and Rio Grande do Norte States.

by Kenneth H. Dinnon III, Emily N. Gallichotte, Ethan J. Fritch, Vineet D. Menachery, Ralph S. Baric

Zika virus (ZIKV) is a mosquito-borne positive sense RNA virus. Recently, ZIKV emerged into the Western hemisphere as a human health threat, with severe disease associated with developmental and neurological complications. The structural envelope protein of ZIKV and other neurotropic flaviviruses contains an extended CD-loop relative to non-neurotropic flaviviruses, and has been shown to augment ZIKV stability and pathogenesis. Here we show that shortening the CD-loop in ZIKV attenuates the virus in mice, by reducing the ability to invade and replicate in the central nervous system. The CD-loop mutation was genetically stable following infection in mice, though secondary site mutations arise adjacent to the CD-loop. Importantly, while shortening of the CD-loop attenuates the virus, the CD-loop mutant maintains antigenicity in immunocompetent mice, eliciting an antibody response that similarly neutralizes both the mutant and wildtype ZIKV. These findings suggest that the extended CD-loop in ZIKV is a determinant of neurotropism and may be a target in live-attenuated vaccine design, for not only ZIKV, but for other neurotropic flaviviruses.

by Felipe Gomes Naveca, Ingra Claro, Marta Giovanetti, Jaqueline Goes de Jesus, Joilson Xavier, Felipe Campos de Melo Iani, Valdinete Alves do Nascimento, Victor Costa de Souza, Paola Paz Silveira, José Lourenço, Mauricio Santillana, Moritz U. G. Kraemer, Josh Quick, Sarah C. Hill, Julien Thézé, Rodrigo Dias de Oliveira Carvalho, Vasco Azevedo, Flavia Cristina da Silva Salles, Márcio Roberto Teixeira Nunes, Poliana da Silva Lemos, Darlan da Silva Candido, Glauco de Carvalho Pereira, Marluce Aparecida Assunção Oliveira, Cátia Alexandra Ribeiro Meneses, Rodrigo Melo Maito, Claudeth Rocha Santa Brígida Cunha, Daniela Palha de Sousa Campos, Marcia da Costa Castilho, Thalita Caroline da Silva Siqueira, Tiza Matos Terra, Carlos F. Campelo de Albuquerque, Laura Nogueira da Cruz, André Luis de Abreu, Divino Valerio Martins, Daniele Silva de Moraes Vanlume Simoes, Renato Santana de Aguiar, Sérgio Luiz Bessa Luz, Nicholas Loman, Oliver G. Pybus, Ester C. Sabino, Osnei Okumoto, Luiz Carlos Junior Alcantara, Nuno Rodrigues Faria
Background Since its first detection in the Caribbean in late 2013, chikungunya virus (CHIKV) has affected 51 countries in the Americas. The CHIKV epidemic in the Americas was caused by the CHIKV-Asian genotype. In August 2014, local transmission of the CHIKV-Asian genotype was detected in the Brazilian Amazon region. However, a distinct lineage, the CHIKV-East-Central-South-America (ECSA)-genotype, was detected nearly simultaneously in Feira de Santana, Bahia state, northeast Brazil. The genomic diversity and the dynamics of CHIKV in the Brazilian Amazon region remains poorly understood despite its importance to better understand the epidemiological spread and public health impact of CHIKV in the country. Methodology/Principal findings We report a large CHIKV outbreak (5,928 notified cases between August 2014 and August 2018) in Boa vista municipality, capital city of Roraima’s state, located in the Brazilian Amazon region. We generated 20 novel CHIKV-ECSA genomes from the Brazilian Amazon region using MinION portable genome sequencing. Phylogenetic analyses revealed that despite an early introduction of the Asian genotype in 2015 in Roraima, the large CHIKV outbreak in 2017 in Boa Vista was caused by an ECSA-lineage most likely introduced from northeastern Brazil. Epidemiological analyses suggest a basic reproductive number of R0 of 1.66, which translates in an estimated 39 (95% CI: 36 to 45) % of Roraima’s population infected with CHIKV-ECSA. Finally, we find a strong association between Google search activity and the local laboratory-confirmed CHIKV cases in Roraima. Conclusions/Significance This study highlights the potential of combining traditional surveillance with portable genome sequencing technologies and digital epidemiology to inform public health surveillance in the Amazon region. Our data reveal a large CHIKV-ECSA outbreak in Boa Vista, limited potential for future CHIKV outbreaks, and indicate a replacement of the Asian genotype by the ECSA genotype in the Amazon region.

by Mariana Kikuti, Jaqueline S. Cruz, Moreno S. Rodrigues, Aline S. Tavares, Igor A. D. Paploski, Monaise M. O. Silva, Perla M. Santana, Laura B. Tauro, Greice A. O. F. Silva, Gúbio S. Campos, Josélio M. G. Araújo, Uriel Kitron, Mitermayer G. Reis, Guilherme S. Ribeiro
Background Rapid diagnosis tests (RDTs) are easy to carry out, provide fast results, and could potentially guide medical treatment decisions. We investigated the performance of a commercially available RDT, which simultaneously detects the non-structural 1 (NS1) dengue virus (DENV) antigen, and IgM and IgG DENV antibodies, using representative serum samples from individuals in a dengue endemic area in Salvador, Brazil. Methodology/Principal findings We evaluated the accuracy of the SD BIOLINE Dengue Duo RDT (Abbott, Santa Clara, USA; former Alere Inc, Waltham, USA) in a random collection of sera. Samples included acute-phase sera from 246 laboratory-confirmed dengue cases and 108 non-dengue febrile patients enrolled in a surveillance study for dengue detection, 73 healthy controls living in the same surveillance community, and 73 blood donors. RDT accuracy was blindly assessed based on the combined results for the NS1 and the IgM test components. The RDT sensitivity was 46.8% (38.6% for the NS1 component and 13.8% for the IgM component). Sensitivity was greater for samples obtained from patients with secondary DENV infections (49.8%) compared to primary infections (31.1%) (P: 0.02) and was also influenced by the result in the confirmatory dengue diagnostic test, ranging from 39.7% for samples of cases confirmed by IgM-ELISA seroconversion between paired samples to 90.4% for samples of cases confirmed by a positive NS1-ELISA. The RDT specificity was 94.4% for non-dengue febrile patients, 87.7% for the community healthy controls, and 95.9% for the blood donors. Conclusions/Significance The SD BIOLINE Dengue Duo RDT showed good specificities, but low sensitivity, suggesting that it may be more useful to rule in than to rule out a dengue diagnosis in dengue endemic regions.

by Jiaqi Cao, Hong Deng, Lei Ye, Xuezheng Ma, Shuru Chen, Xiaohong Sun, Xuemin Wu, Tao Yan, Liping Zhang, Lijuan Liu, Lili Li, Wuping Li, Kongxin Hu

Dengue virus (DENV), a single-stranded RNA virus and Flaviviridae family member, is transmitted by Aedes aegypti and Aedes albopictus mosquitoes. DENV causes dengue fever, which may progress to severe dengue. Hospital-based surveillance was performed in two Chinese regions, Guangzhou and Xishuangbanna, during the dengue epidemics in 2014 and 2015, respectively. Acute-phase serum was obtained from 133 patients with suspected dengue infections during the peak season for dengue cases. Viremia levels, virus sero-positivity, serotype distribution, infection type, clinical manifestations and virus phylogenetics were investigated. Of the 112 DENV-confirmed cases, 92(82.14%) were IgM antibody-positive for DENV, and 69(51.88%) were positive for DENV RNA. From these cases, 47(41.96%) were classified as primary infections, 39(34.82%) as secondary infections and 26 (23.21%) as undetermined infections. The viremia levels were negatively correlated with IgM presence, but had no relationship with the infection type. DENV-1 genotype V dominated in Guangzhou, whereas the DENV-2 Cosmopolitan genotype dominated in Xishuangbanna, where fewer DENV-1 genotype I cases occurred. DENV-2 is associated with severe dengue illness with more serious clinical issues. The strains isolated during 2014–2015 are closely related to the isolates obtained from other Chinese regions and to those isolated recently in Southeast Asian countries. Our results indicate that DENV is no longer an imported virus and is now endemic in China. An extensive seroepidemiological study of DENV and the implementation of vector control measures against it are now warranted in China.

Title: Arbovirus investigation in patients from Mato Grosso during Zika andChikungunya virus introdution in Brazil, 2015–2016
Authors: Costa, M. C. de Souza; Maia, L. M. Siqueira; Souza, Valdinete Costa de; Gonzaga, A. M.; Azevedo, V. Correa de; Martins, L. Ramos; Pavoni, J. H. Chavez; Naveca, Felipe Gomes; Slhessarenko, R. Dezengrini

by Chunling Zou, Chenxiao Huang, Jinyu Zhang, Qihan Wu, Xiaohua Ni, Jiufeng Sun, Jianfeng Dai

Dengue virus (DENV) is the most important vector-borne virus globally. The safe and effective vaccines are still under development and there are no antiviral drugs for DENV induced diseases. In this study, we obtained five DENV1 isolates (DENV1 A to E) from the outbreak of dengue fever in 2014 of Guangzhou, China, and analyzed their replication efficiency and virulence in vitro and in vivo. The results suggested that among the five DENV1 strains, DENV1 B has the highest replication efficiency in both human and mosquito cells in vitro, also causes the highest mortality to suckling mice. Further study suggested that nonstructural proteins from DENV1B have higher capacity to suppress host interferon signaling. In addition, the NS2B3 protease from DENV1B has higher enzymatic activity compared with that from DENV1 E. Finally, we identified that the 64th amino acid of NS2A and the 55th amino acid of NS2B were two virulence determining sites for DENV1. This study provided new evidences of the molecular mechanisms of DENV virulence.

Title: Extract from Aphloia theiformis, an edible indigenous plant from Reunion Island, impairs Zika virus attachment to the host cell surface
Authors: Clain, Elodie; Sinigaglia, Laura; Koishi, Andrea Cristine; Gorgette, Olivier; Gadea, Gilles; Viranaicken, Wildriss; Krejbich-Trotot, Pascale; Mavingui, Patrick; Desprès, Philippe; Santos, Claudia Nunes Duarte dos; Guiraud, Pascale; Jouvenet, Nolwenn; El-Kalamouni, Chaker

by Jung-Seok Lee, Vittal Mogasale, Jacqueline K. Lim, Sowath Ly, Kang Sung Lee, Sopheak Sorn, Esther Andia, Mabel Carabali, Suk Namkung, Sl-Ki Lim, Valéry Ridde, Sammy M. Njenga, Seydou Yaro, In-Kyu Yoon
Background Dengue fever is a rapidly growing public health problem in many parts of the tropics and sub-tropics in the world. While there are existing studies on the economic burden of dengue fever in some of dengue-endemic countries, cost components are often not standardized, making cross-country comparisons challenging. Furthermore, no such studies have been available in Africa. Methods/Principal findings A patient-specific survey questionnaire was developed and applied in Burkina Faso, Kenya, and Cambodia in a standardized format. Multiple interviews were carried out in order to capture the entire cost incurred during the period of dengue illness. Both private (patient’s out-of-pocket) and public (non-private) expenditure were accessed to understand how the economic burden of dengue is distributed between private and non-private payers.A substantial number of dengue-confirmed patients were identified in all three countries: 414 in Burkina Faso, 149 in Kenya, and 254 in Cambodia. The average cost of illness for dengue fever was $26 (95% CI $23-$29) and $134 (95% CI $119-$152) per inpatient in Burkina Faso and Cambodia, respectively. In the case of outpatients, the average economic burden per episode was $13 (95% CI $23-$29) in Burkina Faso and $23 (95% CI $19-$28) in Kenya. Compared to Cambodia, public contributions were trivial in Burkina Faso and Kenya, reflecting that a majority of medical costs had to be directly borne by patients in the two countries. Conclusions/Significance The cost of illness for dengue fever is significant in the three countries. In particular, the current study sheds light on the potential economic burden of the disease in Burkina Faso and Kenya where existing evidence is sparse in the context of dengue fever, and underscores the need to achieve Universal Health Coverage. Given the availability of the current (CYD-TDV) and second-generation dengue vaccines in the near future, our study outcomes can be used to guide decision makers in setting health policy priorities.

by Mark Gallivan, Ben Oppenheim, Nita K. Madhav

Zika virus infection during pregnancy can cause microcephaly and other birth defects. We hypothesized that the Latin America Zika epidemic resulted in pregnant women and their partners adopting behavioral changes to limit risk, leading them to forego travel to Zika-affected locations. We evaluated this hypothesis by studying travelers’ intent and behavior through Twitter data related to babymoon: a holiday taken by parents-to-be before their baby is born. We found the odds of mentioning representative Zika-affected locations in #babymoon tweets dropped significantly (Odds ratio: 0.29, 95% CI: 0.20–0.40) after the Zika-microcephaly association became well-known. This result was further corroborated through a content analysis of #babymoon tweets mentioning Zika-affected locations, which identified if the Twitter user was physically present in the Zika-affected locations. Conversely, we found a small but statistically insignificant increase in the odds of mentioning Zika-free locations from #babymoon tweets (Odds Ratio: 1.11, 95% CI: 0.97–1.27) after the Zika-microcephaly association became well-known.

by J. Guillermo Bond, Adriana R. Osorio, Nancy Avila, Yeudiel Gómez-Simuta, Carlos F. Marina, Ildefonso Fernández-Salas, Pablo Liedo, Ariane Dor, Danilo O. Carvalho, Kostas Bourtzis, Trevor Williams

The sterile insect technique (SIT) may offer a means to control the transmission of mosquito borne diseases. SIT involves the release of male insects that have been sterilized by exposure to ionizing radiation. We determined the effects of different doses of radiation on the survival and reproductive capacity of local strains of Aedes aegypti and Ae. albopictus in southern Mexico. The survival of irradiated pupae was invariably greater than 90% and did not differ significantly in either sex for either species. Irradiation had no significant adverse effects on the flight ability (capacity to fly out of a test device) of male mosquitoes, which consistently exceeded 91% in Ae. aegypti and 96% in Ae. albopictus. The average number of eggs laid per female was significantly reduced in Ae. aegypti at doses of 15 and 30 Gy and no eggs were laid by females that had been exposed to 50 Gy. Similarly, in Ae. albopictus, egg production was reduced at doses of 15 and 25 Gy and was eliminated at 35 Gy. In Ae. aegypti, fertility in males was eliminated at 70 Gy and was eliminated at 30 Gy in females, whereas in Ae. albopictus, the fertility of males that mated with untreated females was almost zero (0.1%) in the 50 Gy treatment and female fertility was eliminated at 35 Gy. Irradiation treatments resulted in reduced ovary length and fewer follicles in both species. The adult median survival time of both species was reduced by irradiation in a dose-dependent manner. However, sterilizing doses of 35 Gy and 50 Gy resulted in little reduction in survival times of males of Ae. albopictus and Ae. aegypti, respectively, indicating that these doses should be suitable for future evaluations of SIT-based control of these species. The results of the present study will be applied to studies of male sexual competitiveness and to stepwise evaluations of the sterile insect technique for population suppression of these vectors in Mexico.

Title: Experimental Adaptation of the Yellow Fever Virus to the Mosquito Aedes albopictus and Potential risk of urban epidemics in Brazil, South America
Authors: Amraoui, Fadila; Pain, Adrien; Piorkowski, Géraldine; Vazeille, Marie; Lima, Dinair Couto; Lamballerie, Xavier de; Oliveira, Ricardo Lourenço de; Failloux, Anna-Bella

by Fadila Amraoui, Wiem Ben Ayed, Yoann Madec, Chafika Faraj, Oumnia Himmi, Ameur Btissam, Mhammed Sarih, Anna-Bella Failloux

In 2015, the mosquito Aedes albopictus was detected in Rabat, Morocco. This invasive species can be involved in the transmission of more than 25 arboviruses. It is known that each combination of mosquito population and virus genotype leads to a specific interaction that can shape the outcome of infection. Testing the vector competence of local mosquitoes is therefore a prerequisite to assess the risks of emergence. A field-collected strain of Ae. albopictus from Morocco was experimentally infected with dengue (DENV), chikungunya (CHIKV), zika (ZIKV) and yellow fever (YFV) viruses. We found that this species can highly transmit CHIKV and to a lesser extent, DENV, ZIKV and YFV. Viruses can be detected in mosquito saliva at day 3 (CHIKV), day 14 (DENV and YFV), and day 21 (ZIKV) post-infection. These results suggest that the local transmission of these four arboviruses by Ae. albopictus newly introduced in Morocco is a likely scenario.
Trial registration: ClinicalTrials.gov APAFIS#6573-201606l412077987v2.

by Alioune Gaye, Eryu Wang, Nikos Vasilakis, Hilda Guzman, Diawo Diallo, Cheikh Talla, Yamar Ba, Ibrahima Dia, Scott C. Weaver, Mawlouth Diallo

Dengue fever (DEN) is the most common arboviral disease in the world and dengue virus (DENV) causes 390 million annual infections around the world, of which 240 million are inapparent and 96 million are symptomatic. During the past decade a changing epidemiological pattern has been observed in Africa, with DEN outbreaks reported in all regions. In Senegal, all DENV serotypes have been reported. These important changes in the epidemiological profile of DEN are occurring in a context where there is no qualified vaccine against DEN. Further there is significant gap of knowledge on the vector bionomics and transmission dynamics in the African region to effectively prevent and control epidemics. Except for DENV-2, few studies have been performed with serotypes 1, 3, and 4, so this study was undertaken to fill out this gap. We assessed the vector competence of Aedes (Diceromyia) furcifer, Ae. (Diceromyia) taylori, Ae. (Stegomyia) luteocephalus, sylvatic and urban Ae. (Stegomyia) aegypti populations from Senegal for DENV-1, DENV-3 and DENV-4 using experimental oral infection. Whole bodies and wings/legs were tested for DENV presence by cell culture assays and saliva samples were tested by real time RT-PCR to estimate infection, disseminated infection and transmission rates. Our results revealed a low capacity of sylvatic and urban Aedes mosquitoes from Senegal to transmit DENV-1, DENV-3 and DENV-4 and an impact of infection on their mortality. The highest potential transmission rate was 20% despite the high susceptibility and disseminated infection rates up to 93.7% for the 3 Ae. aegypti populations tested, and 84.6% for the sylvatic vectors Ae. furcifer, Ae. taylori and Ae. luteocephalus.

by Kanya C. Long, Juan Sulca, Isabel Bazan, Helvio Astete, Hugo L. Jaba, Crystyan Siles, Claudine Kocher, Stalin Vilcarromero, Julia Schwarz, Karin S. Escobedo-Vargas, Fanny Castro-Llanos, Leslye Angulo, Guadalupe Flores, Cesar Ramal-Asayag, Eric S. Halsey, Robert D. Hontz, Valerie A. Paz-Soldan, Thomas W. Scott, Louis Lambrechts, Amy C. Morrison
Background Transmission of dengue virus (DENV) from humans to mosquitoes represents a critical component of dengue epidemiology. Examinations of this process have generally been hampered by a lack of methods that adequately represent natural acquisition of DENV by mosquitoes from humans. In this study, we assessed artificial and natural blood feeding methods based on rates of DENV infection and dissemination within mosquitoes for use in a field-based epidemiological cohort study in Iquitos, Peru. Methodology/Principal findings Our study was implemented, stepwise, between 2011 and 2015. Participants who were 5 years and older with 5 or fewer days of fever were enrolled from ongoing clinic- and neighborhood-based studies on dengue in Iquitos. Wild type, laboratory-reared Aedes aegypti were fed directly on febrile individuals or on blood collected from participants that was either untreated or treated with EDTA. Mosquitoes were tested after approximately 14 days of extrinsic incubation for DENV infection and dissemination. A total of 58 participants, with viremias ranging from 1.3 × 102 to 2.9 × 106 focus-forming units per mL of serum, participated in one or more feeding methods. DENV infection and dissemination rates were not significantly different following direct and indirect-EDTA feeding; however, they were significantly lower for mosquitoes that fed indirectly on blood with no additive. Relative to direct feeding, infection rates showed greater variation following indirect-EDTA than indirect-no additive feeding. Dissemination rates were correlated across all feeding methods. No differences were detected in DENV infection or dissemination rates in mosquitoes fed directly on participants with different dengue illness severity. Conclusions/Significance Our study demonstrates the feasibility of using direct and indirect feeding methods for field-based studies on vector competence. Direct mosquito feeding is preferable in terms of logistical ease, biosecurity, and reliability.

by Amy C. Morrison, Julia Schwarz, Kanya C. Long, Jhonny Cordova, Jennifer E. Rios, W. Lorena Quiroz, S. Alfonso Vizcarra, Robert D. Hontz, Thomas W. Scott, Louis Lambrechts, Valerie A. Paz Soldan

As part of a study to investigate drivers of dengue virus (DENV) transmission dynamics, this qualitative study explored whether DENV-infected residents of Iquitos, Peru, considered it acceptable (1) to participate in direct mosquito feeding experiments (lab-reared Aedes aegypti mosquitoes fed directly on human volunteers) and (2) to provide blood meals indirectly (Ae. aegypti fed on blood drawn from participants by venipuncture). Twelve focus group discussions (FGDs; 94 participants: 82 females and 12 males) were conducted in January 2014 to explore six themes: (1) concerns and preferences regarding direct mosquito feeds and blood draws, (2) comprehension of and misconceptions about study procedures, (3) motivating factors for participation, (4) acceptability of children’s participation, (5) willingness to provide multiple samples over several days, and (6) preference for direct feedings in homes versus the study laboratory. Results of FGDs, including one with 5 of 53 past direct mosquito feed participants, indicated that mosquito feeding procedures are acceptable to Iquitos residents when they are provided with information and a few key messages are properly reinforced. FGD participants’ concerns focused primarily on safety issues rather than discomfort associated with mosquito bites. A video explaining the study dramatically increased comprehension of the study procedures. The majority of participants expressed a preference for mosquito feeding over venipuncture. Adults supported child participation if the children themselves assented. For most participants, home feedings were preferred over those in a laboratory. A major impetus for participation was the idea that results would contribute to an improved understanding of DENV transmission in Iquitos. Findings from our study will support future large-scale studies that employ direct mosquito feeding, a low-risk, non-invasive procedure that is experimentally superior to artificial mosquito feeding methods.

by Jose Angel Regla-Nava, Karla M. Viramontes, Teodora Vozdolska, Anh-Thy Huynh, Tom Villani, Graeme Gardner, Michael Johnson, Pamela J. Ferro, Sujan Shresta, Kenneth Kim

Clinical reports of Zika Virus (ZIKV) RNA detection in breast milk have been described, but evidence conflicts as to whether this RNA represents infectious virus. We infected post-parturient AG129 murine dams deficient in type I and II interferon receptors with ZIKV. ZIKV RNA was detected in pup stomach milk clots (SMC) as early as 1 day post maternal infection (dpi) and persisted as late as 7 dpi. In mammary tissues, ZIKV replication was demonstrated by immunohistochemistry in multiple cell types including cells morphologically consistent with myoepithelial cells. No mastitis was seen histopathologically. In the SMC and tissues of the nursing pups, no infectious virus was detected via focus forming assay. However, serial passages of fresh milk supernatant yielded infectious virus, and immunohistochemistry showed ZIKV replication protein associated with degraded cells in SMC. These results suggest that breast milk may contain infectious ZIKV. However, breast milk transmission (BMT) does not occur in this mouse strain that is highly sensitive to ZIKV infection. These results suggest a low risk for breast milk transmission of ZIKV, and provide a platform for investigating ZIKV entry into milk and mechanisms which may prevent or permit BMT.

Title: Concomitant transmission of dengue, chikungunya and Zika viruses in Brazil: Clinical and epidemiological findings from surveillance for acute febrile illness
Authors: Silva, Monaíse Madalena Oliveira; Tauro, Laura Beatriz; Kikuti, Mariana; Anjos, Rosângela Oliveira; Santos, Viviane Corrêa; Gonçalves, Thaiza Soares Ferreira; Paploski, Igor Adolfho Dexheimer; Moreira, Patrícia Sousa dos Santos; Nascimento, Leile Camila Jacob; Campos, Gúbio Soares; Ko, Albert Icksang; Weaver, Scott C; Reis, Mitermayer Galvão dos; Kitron, Uriel; Ribeiro, Guilherme de Sousa

by Julia Ledien, Kimsan Souv, Rithea Leang, Rekol Huy, Anthony Cousien, Muslim Peas, Yves Froehlich, Raphaël Duboz, Sivuth Ong, Veasna Duong, Philippe Buchy, Philippe Dussart, Arnaud Tarantola

Dengue is a national priority disease in Cambodia. The Cambodian National Dengue Surveillance System is based on passive surveillance of dengue-like inpatients reported by public hospitals and on a sentinel, pediatric hospital-based active surveillance system. This system works well to assess trends but the sensitivity of the early warning and time-lag to usefully inform hospitals can be improved. During The ECOnomic development, ECOsystem MOdifications, and emerging infectious diseases Risk Evaluation (ECOMORE) project’s knowledge translation platforms, Cambodian hospital staff requested an early warning tool to prepare for major outbreaks. Our objective was therefore to find adapted tools to improve the early warning system and preparedness. Dengue data was provided by the National Dengue Control Program (NDCP) and are routinely obtained through passive surveillance. The data were analyzed at the provincial level for eight Cambodian provinces during 2008–2015. The R surveillance package was used for the analysis. We evaluated the effectiveness of Bayesian algorithms to detect outbreaks using count data series, comparing the current count to an expected distribution obtained from observations of past years. The analyses bore on 78,759 patients with dengue-like syndromes. The algorithm maximizing sensitivity and specificity for the detection of major dengue outbreaks was selected in each province. The overall sensitivity and specificity were 73% and 97%, respectively, for the detection of significant outbreaks during 2008–2015. Depending on the province, sensitivity and specificity ranged from 50% to 100% and 75% to 100%, respectively. The final algorithm meets clinicians’ and decisionmakers’ needs, is cost-free and is easy to implement at the provincial level.

by Cécile Beck, Isabelle Leparc-Goffart, Denise Desoutter, Estelle Debergé, Hervé Bichet, Steeve Lowenski, Marine Dumarest, Gaelle Gonzalez, Camille Migné, Jessica Vanhomwegen, Stéphan Zientara, Benoit Durand, Sylvie Lecollinet

New Caledonia and French Polynesia are areas in which arboviruses circulate extensively. A large serological survey among horses from New Caledonia and French Polynesia was carried out to investigate the seroprevalence of flaviviruses in the horse population. Here, 293 equine sera samples were screened for flaviviruses using a competitive enzyme-linked immunosorbent assay (cELISA). The positive sera were then confirmed using a flavivirus-specific microsphere immunoassay (MIA) and seroneutralization tests. This serosurvey showed that 16.6% (27/163) and 30.8% (40/130) of horses were positive for cELISA tests in New Caledonia and French Polynesia, respectively, but the MIA technique, targeting only flaviviruses causing neuro-invasive infections in humans and horses (i.e. West Nile virus [WNV], Japanese encephalitis virus [JEV] and tick-borne encephalitis virus [TBEV]), showed negative results for more than 85% (57/67) of the cELISA-positive animals. Seroneutralization tests with the main flaviviruses circulating in the South Pacific revealed that 6.1% (10/163; confidence interval [95% CI] 3.0%-11.0%) of sera in New Caledonia and 7.7% (10/130; 95% CI 3.8%-13.7%) in French Polynesia were positive for dengue virus serotype 1 (DENV1) and 4.3% (7/163; 95% CI 1.7%-8.6%) in New Caledonia and 15.4% (20/130, 95% CI 9.7%-22.8%) in French Polynesia were found positive for Zika virus (ZIKV). Seroprevalence of the JEV and WNV flaviviruses on the 293 samples from both island groups were comparatively much lower (less than 2%). This seroprevalence study in the horse population shows that horses can be infected with dengue and Zika viruses and that these infections lead to seroconversions in horses. The consequences of these infections in horses and their role in ZIKV and DENV epidemiological cycles are two issues that deserve further investigation.

by Ashley Malmlov, Collin Bantle, Tawfik Aboellail, Kaitlyn Wagner, Corey L. Campbell, Miles Eckley, Nunya Chotiwan, Rebecca C. Gullberg, Rushika Perera, Ronald Tjalkens, Tony Schountz

The emergence of Zika virus (ZIKV) in the New World has led to more than 200,000 human infections. Perinatal infection can cause severe neurological complications, including fetal and neonatal microcephaly, and in adults there is an association with Guillain-Barré syndrome (GBS). ZIKV is transmitted to humans by Aedes sp. mosquitoes, yet little is known about its enzootic cycle in which transmission is thought to occur between arboreal Aedes sp. mosquitos and non-human primates. In the 1950s and ‘60s, several bat species were shown to be naturally and experimentally susceptible to ZIKV with acute viremia and seroconversion, and some developed neurological disease with viral antigen detected the brain. Because of ZIKV emergence in the Americas, we sought to determine susceptibility of Jamaican fruit bats (Artibeus jamaicensis), one of the most common bats in the New World. Bats were inoculated with ZIKV PRVABC59 but did not show signs of disease. Bats held to 28 days post-inoculation (PI) had detectable antibody by ELISA and viral RNA was detected by qRT-PCR in the brain, saliva and urine in some of the bats. Immunoreactivity using polyclonal anti-ZIKV antibody was detected in testes, brain, lung and salivary glands plus scrotal skin. Tropism for mononuclear cells, including macrophages/microglia and fibroblasts, was seen in the aforementioned organs in addition to testicular Leydig cells. The virus likely localized to the brain via infection of Iba1+ macrophage/microglial cells. Jamaican fruit bats, therefore, may be a useful animal model for the study of ZIKV infection. This work also raises the possibility that bats may have a role in Zika virus ecology in endemic regions, and that ZIKV may pose a wildlife disease threat to bat populations.