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This study aimed to evaluate the influence of different anthropic landscape profiles on the diversity and distribution of mosquito species in a rural settlement of the Brazilian Amazon. Eight field collections were conducted at 18 sampling points interspersed throughout 2020-2021. Plastic containers, bamboo internodes, and tires were used as traps to capture immature mosquitoes in three distinct habitats: forest, forest edge, and peridomicile. A total of 15,547 individuals, distributed in 26 species of culicids, were collected. The most abundant species were Culex urichii (8,376 specimens), Culex (Melanoconion) (2,473 specimens), and Aedes albopictus (1,252 specimens). Forest habitat showed the highest abundance, and forest edge showed the highest species richness. Different types of environments influenced both the abundance and richness of mosquitoes. The species composition was also significantly different between the analyzed sites, mainly between forest and peridomicile environments. The change in species dominance could largely explain this change in mosquito community composition. Haemagogus janthinomys, an important sylvatic arbovirus vector, was found in peridomicile habitats and Ae. albopictus, a vector associated with human environments, was found in forest habitats, thus providing evidence of species spillover. Our results indicated that landscape changes affect mosquito communities, influencing their richness and abundance. These changes may have implications for future arboviral outbreaks in this rural settlement due to the possible establishment of sylvatic vector species in anthropic environments.

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INTRODUCTION: Overcoming misinformation is essential considering stigma and discrimination in the HIV/AIDS epidemic. This report presents the preliminary results of a health education strategy based on the massive open online course (MOOC) on Zero Discrimination in Brazil. METHODS: Case study describing the development of the MOOC and its validation using the Delphi technique. Pre- and posttests were administered. People who enrolled from October 2021 to March 2022 were included in the study. RESULTS AND DISCUSSION: MOOC was made available free of charge for mobile phones, tablets, and desktops and included a 90-hr study certificate. Over 6 months, there were n = 665 people enrolled from different regions, mainly from the health field or working in the public health system. The completion rate of people included in the study (26.62%; n = 177) was above the average for other MOOCs. CONCLUSIONS: Initial results are promising but demand more extensive monitoring.

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In the fight against malaria, the key is early treatment with antimalarial chemotherapy, such as artemisinin-based combination treatments (ACTs). However, Plasmodium has acquired multidrug resistance, including the emergence of P. falciparum strains with resistance to ACT. The development of novel antimalarial molecules, that are capable of interfering in the asexual and sexual blood stages, is important to slow down the transmission in endemic areas. In this work, we studied the ability of the mettalo copper-cinchonine complex to interfere in the sexual and asexual stages of Plasmodium. The tested compound in the in vitro assay was a cinchonine derivative, named CinCu (Bis[Cinchoninium Tetrachlorocuprate(II)]trihydrate). Its biological functions were assessed by antiplasmodial activity in vitro against chloroquine-resistant P. falciparum W2 strain. The mice model of P. berghei ANKA infection was used to analyze the antimalarial activity of CinCu and chloroquine and their acute toxicity. The oocyst formation-blocking assay was performed by experimental infection of Anopheles aquasalis with P. vivax infected blood, which was treated with different concentrations of CinCu, cinchonine, and primaquine. We found that CinCu was able to suppress as high as 81.58% of parasitemia in vitro, being considered a molecule with high antiplasmodial activity and low toxicity. The in vivo analysis showed that CinCu suppressed parasitemia at 34% up to 87.19%, being a partially active molecule against the blood-stage forms of P. berghei ANKA, without inducing severe clinical signs in the treated groups. The transmission-blocking assay revealed that both cinchonine and primaquine were able to reduce the infection intensity of P. vivax in A. aquasalis, leading to a decrease in the number of oocysts recovered from the mosquitoes' midgut. Regarding the effect of CinCu, the copper-complex was not able to induce inhibition of P. vivax infection; however, it was able to induce an important reduction in the intensity of oocyst formation by about 2.4 times. It is plausible that the metallo-compound also be able to interfere with the differentiation of parasite stages and/or ookinete-secreted chitinase into the peritrophic matrix of mosquitoes, promoting a reduction in the number of oocysts formed. Taken together, the results suggest that this compound is promising as a prototype for the development of new antimalarial drugs. Furthermore, our study can draw a new pathway for repositioning already-known antimalarial drugs by editing their chemical structure to improve the antimalarial activity against the asexual and sexual stages of the parasite.

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Anopheles darlingi is the main malarial vector in the Brazilian Amazon region. An. nuneztovari s.l., An. triannulatus s.l., An. evansae, and An. benarrochi s.l. do not have a defined role as malarial vectors, although they have been found to be naturally infected with Plasmodium vivax, and some develop oocysts. In this study, we evaluated the importance of low numbers of oocysts in sporozoite salivary gland invasion and transmission. Field-collected mosquitoes were experimentally infected with P. vivax. The infection rates and oocyst and sporozoite infection intensities were evaluated and compared with those of An. aquasalis. We found the highest number of oocysts in An. darlingi (mean = 39.47) and the lowest in An. nuneztovari s.l. (mean = 2). The highest number of sporozoites was observed in An. darlingi (mean = 610) and lowest in An. benarrochi s.l. (mean = 30). Plasmodium vivax DNA was detected in the saliva of all mosquito species after a blood meal. Regardless of the number of oocysts, all species transmitted sporozoites during blood meals. Considering the abundance of these mosquitoes and transmission of sporozoites, it is logical to assume that An. nuneztovari s.l. and An. triannulatus s.l. are involved in the transmission of P. vivax.

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Cutaneous leishmaniasis is endemic in Pernambuco. Aiming to determine the vector species of cutaneous leishmaniasis in an endemic area of the Northeast region of Brazil, this study aimed to use the spatial mapping of human cases of CL and correlate with ecological studies of the vectors in the municipality of Timbaúba, Pernambuco, Brazil. Individuals infected with CL were recruited through active search in their homes and clinically and serologically diagnosed during the period from 2018 to 2019. Sandflies were captured with CDC-type light traps in peridomiciliary environments and these were identified at the species level. Females were separated for DNA extraction and subsequent analysis by PCR. The points of collection of phlebotomes and the residences of individuals with lesions were marked with GPS. During the study period, 60 cases of CL were diagnosed. A higher concentration of CL cases was observed in proximity to Atlantic forest remnants confirmed by heat map. A total of 3744 sandflies was captured and five distinct species were identified, with the predominance of Nyssomyia whitmani. From the females separated for the identification of Leishmania braziliensis DNA, a rate of 0.68% of infected sandflies was obtained. It was concluded that cutaneous leishmaniasis continues to be a rural feature of the area. And from this study, it is concluded that Ny. whitmani is the carrier species of CL in the municipality of Timbaúba, Pernambuco. This is due to abundance in catching, specialization of species and PCR positivity for Leishmania braziliensis.

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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.

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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.

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Mosquito diversity and disease transmission are influenced by landscape modifications, i.e., vectors and pathogens previously found only in forests are now found close to human environments due to anthropic changes. This study determined the diversity and distribution of mosquitoes in forest environments in order to analyze the potential vectors of Amazonian forest arboviruses. Mosquitoes were collected by 1) vertical stratification from forest canopy and ground areas using Hooper Pugedo (HP) light traps and human attraction and 2) horizontal stratification using HP light traps in peridomicile, forest edge, and forest environments near the Rio Pardo rural settlement, Amazonas, Brazil. A total of 3,750 mosquitoes were collected, representing 46 species. 3,139 individuals representing 46 species were sampled by vertical stratification. Both the Shannon-Weaver diversity index (H') and equitability (J') were higher in the canopy than on the ground. 611 individuals representing 13 species were sampled by horizontal stratification. H' decreased in the following order: forest edge > forest > peridomicile, and J' was greater at the forest edge and smaller in the peridomicile environment. Moreover, H' was higher for the human attraction collection method than the HP traps. A total of 671 pools were analyzed by RT-qPCR; three species were positive for Oropouche-like viruses (Ochlerotatus serratus, Psorophora cingulata, and Haemagogus tropicalis) and the minimum infection rate was 0.8%. The composition of mosquito species did not differ significantly between anthropic and forest environments in Rio Pardo. Some mosquito species, due to their abundance, dispersion in the three environments, and record of natural infection, were hypothesized to participate in the arbovirus transmission cycle in this Amazonian rural settlement.

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Experimental studies for understanding the relationship between Plasmodium vivax and its vector hosts are difficult, because of to the lack of a long-term, in vitro continuous culture system unavailability of infected blood samples, seasonality of the disease, and the concentration of most cases in remote areas. This study evaluates the duration of the infectivity of P. vivax to Anopheles aquasalis after collecting blood from malaria-infected patients. Blood was collected from patients and stored at 4 °C and 37 °C. Every day, for 4 days, the blood was fed to An. aquasalis adult females, and a Giemsa-stained thick blood smear was mounted to account for sexual (gametocytes) and asexual (trophozoites and schizonts) stages and calculate parasitemia. Oocysts in the midgut of the mosquitoes were counted on the seventh day after feeding. Kruskal-Wallis test was used to compare the mean number of oocysts (MO) and the parasite density (PD) in each storage condition and post-infection time-points. The Mann-Whitney test was used to compare the number of oocysts for each day between temperatures. The results show that P. vivax stored at 4 °C and at 37 °C has its infectivity to An. aquasalis preserved for 2 days and 3 days, respectively. Infection rate (IR), PD and MO were higher on the day of blood collection and decreased gradually over time. The parasite density (number of parasites/µL) diminished faster at 4 °C than at 37 °C. In this study, a preservation protocol is shown for long-lasting infectivity of P. vivax in a blood sample taken from malaria-infected patients. These results show that infectivity of P. vivax stored at 4 °C and at 37 °C to An. aquasalis persist until 3 days after blood collection, but parasite density, infection rate, and mean of oocysts decreased 24h after blood collection. Since the malaria cases are increasingly far from the urban areas these results indicate that is possible, losing some infectivity, to realize experimental infections several dozen hours after the blood collection. However, it is necessary to improve the procedures for preserving P. vivax gametocytes for mosquito infection in the laboratory.

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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.

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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.

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We describe the immature stages of Migonemyia migonei, which is the vector of Leishmania (Viannia) braziliensis, the etiological agent of cutaneous leishmaniasis in South America, and a putative vector of Leishmania infantum chagasi. Scanning Electron Microscopy (SEM) was used to refine the description of the structures of the egg, all instar larvae, and the pupa. The eggs have polygonal cells on the egg exochorion, and differences between larval and pupal chaetotaxy have been highlighted. Different sensillary subtypes-trichoidea, basiconica, coelonica and campanoformia-were observed in the larval stages. The results presented herein contribute to the taxonomy of Mg. migonei and may contribute to future studies on the phylogeny of this important vector species.

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Anthropogenic environments provide favorable conditions for some species, which is especially true of mosquitoes that present eclecticism at the moment of choice for the site of oviposition. In the present study, the diversity of mosquitoes was assessed by providing plastic containers, bamboo internodes, and tires in a forest, the forest edge, and peridomicile environments in a rural settlement area. Eighteen sampling points were chosen, delimited by a buffer of 200 m, placed in three environments: forest, forest edge, and peridomicile. In each environment, larvitraps were installed, separated by a minimum distance of 7 m and 1 m from the ground. A total of 10,131 immature mosquitoes of 20 species were collected. The most abundant species was Culex urichii (29.5%), followed by Trichoprosopon digitatum (27.1%), and Cx. (Melanoconion) spp. (10.4%). There was a difference in the composition of immature mosquito populations between larvitraps (p < 0.0005), and the plastic container hosted a greater diversity of species, whereas tires presented a greater abundance of individuals. The forest, forest edge, and peridomicile environments were also different with regard to diversity of immature mosquito populations (p < 0.0010). The forest edge was the environment with the greatest diversity of species, followed by the peridomicile and forest environments. In the forest and peridomicile, plastic container larvitraps had the greatest diversity, whereas the forest edge tire presented the largest number of individuals. Further, tire larvitraps collected the largest number of individuals in all environments. Ten species associated with the bamboo internode and tire were identified. The preference of species for artificial larvitraps, such as the plastic container and tire, even in wild environments was noted. These artificial objects may represent a risk factor for the population living in this region, as all vector species found in the study were present in plastic containers and tires.

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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.

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ANTECEDENTES: Se desconoce el contenido de grasas saturada (GS) y trans (Gtrans) en panes sin etiqueta y sin vigilancia por entidad regulatoria de Colombia. PROPÓSITO: Comparar el contenido de GS y Gtrans en panes de algunas panaderías de Medellín con lo reglamentado: 0,5 g/50 g de pan, para ambos tipos de grasa según resolución 2508 de 2012 del Ministerio de Salud. MATERIALES Y MÉTODOS: Se muestrearon 45 panes de tres tipos (aliñado, leche y croissant) en tres niveles socioeconómicos (NSE) según ubicación de la panadería. Se cuantificó grasa total gravimétricamente y GS y Gtrans por cromatografía de gases. RESULTADOS: El porcentaje de GS y Gtrans con respecto a la grasa total extraíble fue 43,8% y 0,6% en pan aliñado, 37,4% y 0,6% en pan leche y 32,2% y 0,9% en pan croissant, sin diferencia significativa entre ellos (p= 0,155 y 0,184, respectivamente). CONCLUSIONES: Comparado con la resolución 2508, el contenido de GS fue mayor al límite establecido en resolución y el contenido de Gtrans fue menor en pan aliñado, leche o croissant. Los panes evaluados en las panaderías del NSE bajo contenían mayor Gtrans que los otros.

BACKGROUND: The content of saturated (GS) and trans (Gtrans) fats in unlabeled and breads that are not regulated by the Colombian government is unknown. PURPOSE: To compare the content of GS and Gtrans in breads of bakeries in Medellín with the following regulation: 0.5 g/50 g of bread, for both types of fat according to the 2012 Ministry of Health resolution #2508. MATERIALS AND METHODS: 45 breads of three types (seasoned, milk and croissant) were sampled at bakeries from three socioeconomic levels (NSE) according to the location. Total fat was quantified gravimetrically and GS and Gtrans by gas chromatography. RESULTS: The percentage of GS and Gtrans with respect to the total extractable fat was 43.8% and 0.6% in seasoned bread, 37.4% and 0.6% in milk bread and 32.2% and 0.9% in croissant bread, with no significant difference by bread type (p= 0.155 and 0.184, respectively). CONCLUSIONS: GS content of sampled breadswas greater than the limit established in the resolution. Gtrans content was lower in seasoned bread andmilk bread compared to croissant. The breads evaluated in the bakeries of lower NSE contained higher Gtrans than the others.

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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.

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Objetivo: Estimar las frecuencias de mutaciones y de polimorfismos adicionales asociados con resistencia a los fármacos inhibidores de la integrasa del virus de inmunodeficiencia humana tipo 1 (VIH-1). Metodología: Estudio descriptivo, de corte transversal, en individuos VIH-1 positivos de la ciudad de Medellín, quienes no habían recibido tratamiento antirretroviral. En ellos se determinó, a través del método de 2-dideoxinucleótidos y el sistema ABI3730XL, la secuencia del gen de la integrasa del VIH-1 a partir del ARN viral circulante, la cual fue analizada en la base de datos de resistencia a medicamentos antirretrovirales de la Universidad de Stanford y según reportes de literatura científica. Resultados: Se encontraron las siguientes mutaciones (con sus respectivas frecuencias): una mutación mayor, E138K (1/46), tres mutaciones accesorias G163E (3/46), L74I (3/50) y E157Q (2/48), una mutación no polimórfica A128T (1/49) y otras dos mutaciones potencialmente asociadas con resistencia a inhibidores de integrasa S230N (9/39) y S119P/R/T (4/47, 2/47 y 14/47, respectivamente). Conclusiones: En las secuencias analizadas, llama la atención la presencia de al menos una mutación asociada a resistencia a inhibidores de integrasa en el 14% de los individuos estudiados, sugiriendo una pobre presión selectiva de este tipo de fármacos en la población viral circulante en la zona.

Aim: To estimate the frequencies of major and accessory mutations, as well as additional polymorphisms associated with resistance to human immunodeficiency virus type 1 (VIH-1) integrase strand transfer inhibitors. Materials and methods: Descriptive cross-sectional study, focused on HIV-1 positive individuals from Medellín, recruited between 2013 and 2015, and that had not received antiretroviral therapy. In these patients, the sequence from HIV-1 integrase was determined from circulating viral RNA through Sanger chain termination method with the ABI3730XL system, and the sequences were analyzed using the HIV Drug Resistance Database from the University of Stanford, together with previous literature reports. Results: The following mutations associated with resistance to integrase strand transfer inhibitors, along with its respective frequencies, were found: one major mutation, E138K (1/46), three accessory mutations, G163E (3/46), L74I (3/50) and E157Q (2/48); one non-polymorphic mutation, A128T (1/49); and two mutations potentially associated with resistance to integrase strand transfer inhibitors, S230N (9/39) and S119P/R/T (4/47, 2/47 and 14/47, respectively). Conclusions: In the sequences analyzed, it is noteworthy the presence of at least one mutation related with resistance to integrase inhibitors in 14% of the studied patients, suggesting a poor selective pressure of this kind of drugs in the circulating viral population in our region.

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Whole mitogenome sequences (mtDNA) have been exploited for insect ecology studies, using them as molecular markers to reconstruct phylogenies, or to infer phylogeographic relationships and gene flow. Recent Anopheles phylogenomic studies have provided information regarding the time of deep lineage divergences within the genus. Here we report the complete 15,393 bp mtDNA sequences of Anopheles aquasalis, a Neotropical human malaria vector. When comparing its structure and base composition with other relevant and available anopheline mitogenomes, high similarity and conserved genomic features were observed. Furthermore, 22 mtDNA sequences comprising anopheline and Dipteran sibling species were analyzed to reconstruct phylogenies and estimate dates of divergence between taxa. Phylogenetic analysis using complete mtDNA sequences suggests that A. aquasalis diverged from the Anopheles albitarsis complex ~28 million years ago (MYA), and ~38 MYA from Anopheles darlingi. Bayesian analysis suggests that the most recent ancestor of Nyssorhynchus and Anopheles + Cellia was extant ~83 MYA, corroborating current estimates of ~79-100 MYA. Additional sampling and publication of African, Asian, and North American anopheline mitogenomes would improve the resolution of the Anopheles phylogeny and clarify early continental dispersal routes.

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INTRODUCTION: Malaria and leishmaniases are transmitted by vectors during blood-feeding. Vector-infected animals develop antibodies against the vector's saliva. This study evaluated IgY antibody detection in the chicken eggs exposed to bites from Migonemyia migonei, Lutzomyia longipalpis and Anopheles aquasalis. METHODS: We used ELISA to quantify the antibody levels in the sera and exposed chicken eggs. RESULTS: High IgY levels were observed following immunization; furthermore, higher reactivity was observed in the eggs and species-specific immune response was observed post final immunization. CONCLUSIONS: Chicken eggs can be used as sentinels to surveil vector saliva antibodies.

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Roads and highways can affect the spread of insect-borne diseases by limiting or amplifying the spatiotemporal distribution of vectors, pathogens, and hosts, which can, in turn, lead to the creation of a nidus of infection. The aim of this study was to compare the diversity (richness and abundance) of phlebotomine sand flies in household and forest edge environments found along two different segments of an Amazonian highway. Sampling was conducted along the northern and southern portions of highway BR-319, in Amazonas State, Brazil. At each sampling point, Hoover Pugedo traps were set in indoor and outdoor habitats, and at forests edges, and captures were made between 06:00 pm and 06:00 am. A total of 1,189 sand flies were captured and 48 species were identified. As expected, a greater number of species and individuals were captured in forest edge environments. Permutational Multivariate Analyses of Variance (PERMANOVA) and Permutational Analyses of Multivariate Dispersions (PERMDISP) analyses showed that sand fly fauna differed significantly among habitats, but no variance in species composition was observed between the two road segments. Some of the captured species were species that have been implicated as vectors of Leishmania spp. Ross, 1903 (Kinetoplastida: Trypanosomatidae).

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