Resumo O gênero Culicoides é um diminuto diptero da família Ceratopogonidae com distribuição mundial. Devido aos seus hábitos hematófagos, as espécies desse gênero são um incômodo para os humanos e animais domésticos transmitem protozoários, vermes filariais, bem como, diversos arbovírus. A região Nordeste do Brasil é caracterizada por clima semiárido e poucos corpos d’água, o que não favorece criadouros de ceratopogonídeos. Além do fator climático, estudos sobre mosquitos picadores são escassos nesta região geográfica. O município de Pacoti, localizado na Serra de Baturité, no Ceará, é uma importante área de produção agrícola e ecoturismo; além disso, Pacoti está localizada na área de preservação ambiental. Devido à importância médica e veterinária desses maruins, foi então realizada uma pesquisa em Pacoti. Para a captura das amostras foram utilizadas armadilhas luminosas do tipo CDC em áreas florestadas is, onde as coletas foram realizadas em dois períodos: março de 2015 e agosto de 2022. Das 17 espécies encontradas nesse estudo, 12 são novos registros para o município de Pacoti, nove são novos registros para o estado do Ceará e quatro são registrados pela primeira vez para a região Nordeste. Das espécies vetoras incriminadas temos: C. paraensis, C. insignis e C. pusillus. O conhecimento da biodiversidade e a atualização da distribuição geográfica dos Culicoides poderão contribuir para medidas de controle de doenças transmitidas por vetores no Ceará. mundial hematófagos protozoários filariais como arbovírus dágua, dágua d água, água d’água ceratopogonídeos climático Baturité ecoturismo disso ambiental maruins is períodos 201 2022 1 estudo temos C paraensis pusillus 20 202 2

Abstract Culicoides is a small dipteran genus of Ceratopogonidae Family with worldwide distribution. Due to their hematophagous habits, species within this genus are a biting nuisance to humans and domestic animals. They also transmit protozoa, filarial worms, and several arboviruses. The Northeastern region of Brazil is characterized by a semi-arid climate and few bodies of water, which does not favor breeding sites for ceratopogonids. In addition to the climatic factor, studies on Biting Midges Culicoides are scarce in this geographic region. The municipality of Pacoti, located in Serra de Baturité, in Ceará, is an important area for agricultural production and ecotourism; also, Pacoti is located in an environmental protection area. Due to the medical and veterinary importance of these biting midges, a survey was conducted in Pacoti. For capture, CDC-type light traps were used near forested areas, where collections were carried out during two periods: March 2015, and August 2022. Of the 17 species found, 12 are new records for the municipality of Pacoti, nine are new records for the state of Ceará, and four are recorded for the first time for the Northeast region. Of the registered incriminated vectors, we found C. paraensis, C. insignis, and C. pusillus. The knowledge of biodiversity and geographic distribution update of Culicoides may contribute to prevention and control measures for vector-borne diseases in Ceará. habits animals protozoa worms arboviruses semiarid semi arid water ceratopogonids factor Baturité Ceará ecotourism midges capture CDCtype CDC type areas periods 2015 2022 1 vectors C paraensis insignis pusillus vectorborne vector borne 201 202 20 2

BACKGROUND Trypanosomatids are widespread and cause diseases - such as trypanosomiasis, sleeping sickness, Chagas disease, and cutaneous and visceral leishmaniasis - in animals and humans. These diseases occur in both rural and urban regions due to unplanned growth and deforestation. Thus, wild and synanthropic reservoir hosts living in residential areas are risk factors. OBJECTIVE We aimed to evaluate the diversity of small mammals (rodents and marsupials), and the occurrence of trypanosomatids, especially Leishmania, in the rural settlement of Presidente Figueiredo, Amazonas. METHODS Animals were collected using Sherman, Tomahawk, and Pitfall traps along 16 trails in four landscapes: continuous forest, forest with planting, planting, and peridomiciliar. Leishmania sp. was detected in liver samples by polymerase chain reaction targeting kDNA. FINDINGS Diversity was higher in forests with planting and lower around residences. In total, 135 mammals (81 rodents and 54 marsupials covering 14 genera) were captured. Rodents presented infection rates (IR) of 74% and marsupials of 48%. Rodents in domicile landscapes presented a higher IR (92.9%), while marsupials showed a higher IR in forests (53.3%). MAIN CONCLUSIONS The results suggest high prevalence of trypanosomatids across 12 mammalian genera possibly involved as reservoir hosts in the enzootic transmission of leishmaniasis in the Amazon’s rural, peridomiciliar landscape.

BACKGROUND Different strategies for improvement of malaria control and elimination are based on the blockage of malaria parasite transmission to the mosquito vector. These strategies include the drugs that target the plasmodial sexual stages in humans and the early developmental stages inside mosquitoes. OBJECTIVES Here we tested Malaria Box compounds in order to evaluate their activity against male and female gametocytes in Plasmodium berghei, mosquito infection in P. vivax and ookinete formation in both species. METHODS/FINDINGS The membrane feeding assay and the development of ookinetes by a 24 h ex vivo culture and the ookinete yield per 1000 erythrocytes were used to test transmission-blocking potential of the Malaria Box compounds in P. vivax. For P. berghei we used flow cytometry to evaluate male and female gametocyte time course and fluorescence microscopy to check the ookinete development. The two species used in this study showed similar results concerning the compounds’ activity against gametocytes and ookinetes, which were different from those in P. falciparum. In addition, from the eight Malaria Box compounds tested in both species, compounds MMV665830, MMV665878 and MMV665941 were selected as a hit compounds due the high inhibition observed. CONCLUSION Our results showed that P. berghei is suitable as an initial screening system to test compounds against P. vivax.

Abstract INTRODUCTION: The Culicoides transmit a variety of pathogens. Our aim was to survey the Culicoides species occurring in an Amazonian rural settlement, comparing abundance, richness, and diversity in different environments. METHODS: Culicoides were captured using CDC light traps. The Shannon-Wiener (H’) and Rényi indices were used to compare species diversity and evenness between environments, the equitability (J’) index was used to calculate the uniformity of distribution among species, and similarity was estimated using the Jaccard similarity index. A permutational multivariate analysis of variance was applied to assess the influence of environment on species composition. A non-metric dimensional scale was used to represent the diversity profiles of each environment in a multidimensional space. RESULTS: 6.078 Culicoides were captured, representing 84 species (45 valid species/39 morphotypes). H’ values showed the following gradient: forest > capoeira > peridomicile > forest edge. The equitability J’ was greater in capoeira and forests compared to peridomiciles and the forest edge. The population compositions of each environment differed statistically, but rarefaction estimates indicate that environments of the same type possessed similar levels of richness. Species of medical and veterinary importance were found primarily in peridomiciles: C. paraensis, vector of Oropouche virus; C. insignis and C. pusillus, vectors of Bluetongue virus; C. filariferus, C. flavivenula, C. foxi, and C. ignacioi, found carrying Leishmania DNA. CONCLUSIONS: This study indicates that diversity was higher in natural environments than in anthropized environments, while abundance and richness were highest in the most anthropized environment. These findings suggest that strictly wild Culicoides can adapt to anthropized environments.

BACKGROUND Aedes aegypti is considered the main Zika virus (ZIKV) vector, and is thought to be responsible for the 2015-2016 outbreak in Brazil. Zika positive Ae. aegypti males collected in the field suggest that vertical and/or venereal transmission of ZIKV may occur. OBJECTIVES In this study, we aimed to demonstrate that venereal transmission of ZIKV by Ae. aegypti can occur under laboratory conditions. METHODS Ae. aegypti collected in the city of Manaus, confirmed as negative for Zika, Dengue and Chikungunya virus by reverse transcription real-time polymerase chain reaction (RT-qPCR) (AaM3V- strain), were reared under laboratory conditions and used for the experiments. The ZIKV used in this study was isolated from a patient presenting with symptoms; ZIKV was confirmed by RT-qPCR. Experiment 1: virgin male mosquitoes of AaM3V- strain were intrathoracically inoculated with a ZIKV suspension; four days after injection, they were transferred to a cage containing virgin females of AaM3V- strain and left to copulate for five days. Experiment 2: virgin female mosquitoes of AaM3V- strain were orally infected with a ZIKV suspension by blood feeding membrane assay; nine days after blood feeding, they were placed in cages with Ae. aegypti AaM3V- virgin males and left to copulate for four days. After copulation, all mosquitoes were individually evaluated for viral infection by RT-qPCR. FINDINGS The mean infection rate in Experiment 1 and Experiment 2 was 45% and 35%, respectively. In both experiments, cycle threshold values ranged from 13 to 35, indicating the presence of viral genomes. MAIN CONCLUSION Ae. aegypti males intrathoracically inoculated with a ZIKV suspension are infected and can transmit the virus to uninfected females by mating. Moreover, Ae. aegypti females orally infected with a ZIKV suspension can transmit the virus to uninfected males by copulation. This study shows that ZIKV infection of Ae. aegypti mosquitoes occurs not only during blood feeding, but also during copulation.