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Mosquitoes are the most important vectors of emerging infectious diseases. During the past decade, our understanding of the diversity of viruses they carry has greatly expanded. Most of these viruses are considered mosquito-specific, but there is increasing evidence that these viruses may affect the vector competence of mosquitoes. Metagenomics approaches have focused on specific mosquito species for the identification of what is called the core virome. Despite the fact that, in most ecosystems, multiple species may participate in virus emergence and circulation, there is a lack of understanding of the virus-carrier/host network for both vector-borne and mosquito-specific viruses. Here, we studied the core virome of mosquitoes in a diverse local ecosystem that had 24 different mosquito species. The analysis of the viromes of these 24 mosquito species resulted in the identification of 34 viruses, which included 15 novel viruses, as determined according to the species demarcation criteria of the respective virus families. Most of the mosquito species had never been analysed previously, and a comparison of the individual viromes of the 24 mosquito species revealed novel relationships among mosquito species and virus families. Groups of related viruses and mosquito species from multiple genera formed a complex web in the local ecosystem. Furthermore, analyses of the virome of mixed-species pools of mosquitoes from representative traps of the local ecosystem showed almost complete overlap with the individual-species viromes identified in the study. Quantitative analysis of viruses' relative abundance revealed a linear relationship to the abundance of the respective carrier/host mosquito species, supporting the theory of a stable core virome in the most abundant species of the local ecosystem. Finally, our study highlights the importance of using a holistic approach to investigating mosquito viromes relationships in rich and diverse ecosystems., (© The Author(s) 2022. Published by Oxford University Press.)

Background: Haiti is one of the Caribbean countries where malaria persists. More than 99% of malaria cases are caused by Plasmodium falciparum, the main vector being the mosquito Anopheles albimanus. In this paper, we describe the epidemiological profile of malaria in Haiti between 2009 and 2018., Methods: We analyzed information on cases reported by the Ministry of Health of Haiti and the World Health Organization (WHO)., Results: Between 2009 and 2018, 232,479 malaria cases were reported by the Ministry of Public Health and Population (MSPP); an increase in the incidence of malaria in the country in 2010, followed by a decrease in 2011, was primarily observed. Due to recent efforts to reduce malaria by 2020, its incidence declined from 60,130 cases in 2010 to 8,978 cases in 2018. Controversially, in terms of the number of reported cases, the MSPP and WHO report conflicting data. However, the results from both datasets present the same trend in Haiti from 2009 to 2018. The results also illustrate the endemicity of the disease throughout Haiti, both in rural and urban areas, especially along the coast., Conclusions: This study emphasizes the need to promote official data collection and analyses, as well as the application of epidemiological surveillance of malaria at the municipal level, for a better understanding of the real impact of malaria on the Haitian population and to create more appropriate interventions.

Identifying the species of the subfamily Anophelinae that are Plasmodium vectors is important to vector and malaria control. Despite the increase in cases, vector mosquitoes remain poorly known in Brazilian indigenous communities. This study explores Anophelinae mosquito diversity in the following areas: (1) a Yanomami reserve in the northwestern Amazon Brazil biome and (2) the Pantanal biome in southwestern Brazil. This is carried out by analyzing cytochrome c oxidase ( COI ) gene data using Refined Single Linkage (RESL), Assemble Species by Automatic Partitioning (ASAP), and tree-based multi-rate Poisson tree processes (mPTP) as species delimitation approaches. A total of 216 specimens collected from the Yanomami and Pantanal regions were sequenced and combined with 547 reference sequences for species delimitation analyses. The mPTP analysis for all sequences resulted in the delimitation of 45 species groups, while the ASAP analysis provided the partition of 48 groups. RESL analysis resulted in 63 operational taxonomic units (OTUs). This study expands our scant knowledge of anopheline species in the Yanomami and Pantanal regions. At least 18 species of Anophelinae mosquitoes were found in these study areas. Additional studies are now required to determine the species that transmit Plasmodium spp. in these regions.

Background: Aedes albopictus is a very invasive mosquito, which has recently colonized tropical and temperate regions worldwide. Of concern is its role in the spread of emerging or re-emerging mosquito-borne diseases. Ae. albopictus from south-western Europe and Brazil were studied to infer genetic and phenetic diversity at intra-individual, intra-population and inter-population levels, and to analyse its spread., Methods: Genotyping was made by rDNA 5.8S-ITS-2 and mtDNA cox1 sequencing to assess haplotype and nucleotide diversity, genetic distances and phylogenetic networks. Male and female phenotyping included combined landmark-and outlined-based geometric morphometrics of wing size and shape., Results: Specimens from seven populations from Spain, France and Brazil provided 12 cox1 and 162 5.8S-ITS-2 haplotypes, with great genetic variability difference between both markers (0.9% vs 31.2%). Five cox1 haplotypes were shared with other countries, mainly Italy, USA and China, but none was shared between Europe and Brazil. The 5.8S-ITS-2 showed 2-7 intra-individual (mean 4.7) and 16-34 intra-/inter-population haplotypes (24.7), including haplotypes shared between Spain, France and Brazil. A 4.3% of ITS-2 haplotypes were shared, mainly with Italy, USA and Thailand, evidencing worldwide spread and introductions from areas where recent outbreaks of Ae. albopictus-transmitted pathogens occurred. Wing size showed sex differences. Wing shape distinguished between Brazilian and European specimens. Both genetic and morphometric markers showed differences between insular Spain and continental Spain, France and Brazil., Conclusions: ITS-2 proves to be a useful marker to assess Ae. albopictus spread, providing pronouncedly more information than cox1, including intra-individual, intra-population and inter-population levels, furnishing a complete overview of the evolutionary exchanges followed by this mosquito. Wing morphometry proves to be a useful phenotyping marker, allowing to distinguish different populations at the level of both male and female specimens. Results indicate the need for periodic surveillance monitorings to verify that no Ae. albopictus with high virus transmission capacity is introduced into Europe.

The oviposition behavior of mosquitoes varies between species. We documented the unusual mechanism of egg laying in the mosquito Sabethes albiprivus Theobald with the aid of high speed video recordings in the laboratory. A sapucaia (Lecythis pisonis Camb.) nut oviposition trap, described herein, was used to simulate a tree hole with a small opening, which is the natural larval habitat of Sa. albiprivus. We showed that females approach the opening and perform a sequence of rapid, short up-and-down flights before egg laying. At this time, the egg is already visible externally, being held at the very tip of the abdomen. Females catapult one egg at a time by rapidly curling their abdomen downward, sending the egg through the opening while their legs are positioned in different configurations throughout the event. The estimated velocity of the catapulted eggs was almost 1 m/s.

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) chikungunya (CHIKV), o'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group has been established to investigate natural history, epidemiology and clinical aspects of infection by these viruses. Here, we present a report dedicated to entomological aspects of CHIKV, ONNV and MAYV. Recent global expansion of chikungunya virus has been possible because CHIKV established a transmission cycle in urban settings using anthropophilic vectors such as Aedes albopictus and Aedes aegypti. MAYV and ONNV have a more limited geographic distribution, being confined to Africa (ONNV) and central-southern America (MAYV). ONNV is probably maintained through an enzootic cycle that has not been characterized yet, with Anopheles species as main vectors and humans as amplification hosts during epidemics. MAYV is transmitted by Haemagogus species in an enzootic cycle using non-human primates as the main amplification and maintenance hosts, and humans becoming sporadically infected when venturing in or nearby forest habitats. Here, we focused on the transmission cycle and natural vectors that sustain circulation of these viruses in their respective locations. The knowledge of the natural ecology of transmission and the capacity of different vectors to transmit these viruses is crucial to understand CHIKV emergence, and to assess the risk that MAYV and ONNV will expand on wide scale using anthropophilic mosquito species not normally considered primary vectors. Finally, the experts identified knowledge gaps and provided adapted recommendations, in order to address future entomological investigations in the right direction., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

BACKGROUND Studies on malaria vectors in the Pantanal biome, Central Brazil, were conducted more than half a century ago. OBJECTIVES To update anopheline records and assess receptivity and vulnerability to malaria transmission. METHODS Five-day anopheline collections were conducted bimonthly in Salobra, Mato Grosso do Sul state, for one year. Indoors, mosquitoes were collected from their resting places, while in open fields, they were captured using protected human-baited and horse-baited traps near the house and at the Miranda River margin, respectively. Hourly biting activity outdoors was also assessed. Secondary data were collected on the arrival of tourists, economic projects, and malaria cases. FINDINGS A total of 24,894 anophelines belonging to 13 species were caught. The main Brazilian malaria vector Anopheles darlingi was the predominant species, followed by An. triannulatus s.l. Hourly variation in anopheline biting showed three main peaks occurring at sunset, around midnight, and at sunrise, the first and last being the most prominent. The highest density of all species was recorded near the river margin and during the transition period between the rainy and early dry seasons. This coincides with the time of main influx of outsider workers and tourists, whose activities mostly occur in the open fields and frequently start before sunrise and last until sunset. Some of these individuals originate from neighbouring malaria-endemic countries and states, and are likely responsible for the recorded imported and introduced malaria cases. MAIN CONCLUSION Pantanal is a malaria-prone area in Brazil. Surveillance and anopheline control measures must be applied to avoid malaria re-emergence in the region.

Here, we describe the development of the MosqTent, an innovative double-chamber mosquito trap in which a human being attracts mosquitoes while is protected from being bitten within the inner chamber of the trap, while mosquitoes are lured to enter an outer chamber where they are trapped. The MosqTent previously collected an average of 3,000 anophelines/man-hour compared to 240 anophelines/man-hour for the human landing catch (HLC), thereby providing high numbers of human host-seeking mosquitoes while protecting the collector from mosquito bites. The MosqTent performed well by collecting a high number of specimens of Anopheles marajoara, a local vector and anthropophilic mosquito species present in high density, but not so well in collecting An. darlingi, an anthropophilic mosquito species considered the main vector in Brazil but is present in low-density conditions in the area. The HLC showed a higher efficiency in collecting An. darlingi in these low-density conditions. The MosqTent is light (<1 kg), portable (comes as a bag with two handles), flexible (can be used with other attractants), adaptable (can be deployed in a variety of environmental settings and weather conditions), and it can be used in the intra-, peri-, and in the extradomicile. Also, the MosqTent collected similar portions of parous females and anthropophilic mosquito species and collects specimens suitable for downstream analysis. Further developments may include testing for other fabric colors, different mesh sizes and dimensions for other hematophagous insects and conditions, additional chemical mosquito attractants, and even the replacement of the human attractant in favor of other attractants. MosqTent modifications that would allow the trap to be applied as a vector control tool with killing action could also be explored.

Here we present the first in a series of articles about the ecology of immature stages of anophelines in the Brazilian Yanomami area. We propose a new larval habitat classification and a new larval sampling methodology. We also report some preliminary results illustrating the applicability of the methodology based on data collected in the Brazilian Amazon rainforest in a longitudinal study of two remote Yanomami communities, Parafuri and Toototobi. In these areas, we mapped and classified 112 natural breeding habitats located in low-order river systems based on their association with river flood pulses, seasonality and exposure to sun. Our classification rendered seven types of larval habitats: lakes associated with the river, which are subdivided into oxbow lakes and nonoxbow lakes, flooded areas associated with the river, flooded areas not associated with the river, rainfall pools, small forest streams, medium forest streams and rivers. The methodology for larval sampling was based on the accurate quantification of the effective breeding area, taking into account the area of the perimeter and subtypes of microenvironments present per larval habitat type using a laser range finder and a small portable inflatable boat. The new classification and new sampling methodology proposed herein may be useful in vector control programs.

Distribution, abundance, feeding behaviour, host preference, parity status and human-biting and infection rates are among the medical entomological parameters evaluated when determining the vector capacity of mosquito species. To evaluate these parameters, mosquitoes must be collected using an appropriate method. Malaria is primarily transmitted by anthropophilic and synanthropic anophelines. Thus, collection methods must result in the identification of the anthropophilic species and efficiently evaluate the parameters involved in malaria transmission dynamics. Consequently, human landing catches would be the most appropriate method if not for their inherent risk. The choice of alternative anopheline collection methods, such as traps, must consider their effectiveness in reproducing the efficiency of human attraction. Collection methods lure mosquitoes by using a mixture of olfactory, visual and thermal cues. Here, we reviewed, classified and compared the efficiency of anopheline collection methods, with an emphasis on Neotropical anthropophilic species, especially Anopheles darlingi, in distinct malaria epidemiological conditions in Brazil.

Roraima is the northernmost state of Brazil, bordering both Venezuela and Guyana. Appropriate climate and vector conditions for dengue transmission together with its proximity to countries where all four dengue serotypes circulate make this state, particularly the capital Boa Vista, strategically important for dengue surveillance in Brazil. Nonetheless, few studies have addressed the population dynamics of Aedes aegypti in Boa Vista. In this study, we report temporal and spatial variations in Ae. aegypti population density using ovitraps in two highly populated neighbourhoods; Centro and Tancredo Neves. In three out of six surveys, Ae. aegypti was present in more than 80% of the sites visited. High presence levels of this mosquito suggest ubiquitous human exposure to the vector, at least during part of the year. The highest infestation rates occurred during the peak of the rainy seasons, but a large presence was also observed during the early dry season (although with more variation among years). Spatial distribution of positive houses changed from a sparse and local pattern to a very dense pattern during the dry-wet season transition. These results suggest that the risk of dengue transmission and the potential for the new serotype invasions are high for most of the year.

Knowledge of vector distribution is important for the design of effective local malaria control programs. Here we apply ecological niche modeling to analyze and predict the distributions of malaria vectors based on entomological collection points in the State of Roraima in the northern Brazilian Amazon Basin. Anopheline collections were conducted from 1999 to 2003 at 76 localities, all with active malaria transmission. A total of 13 anopheline species was identified from 17,074 adult females collected: Anopheles darlingi, An. albitarsis s.l., An. nuneztovari, An. triannulatus s.l., An. braziliensis, An. peryassui, An. oswaldoi s.l., An. mattogrossensis, An. strodei, An. evansae, An. squamifemur, An. mediopunctatus s.l, An. intermedius. Anopheles darlingi, and An. albitarsis were the most frequently found species. An. squamifemur was found for the first time in Roraima. A distributional prediction model (genetic algorithm for rule-set prediction-GARP) and environmental variables were used to predicted potential distribution range for six anopheline species that occurred at > or = 19 collection points. The method allows for the application of moderate sample sizes to produce distribution maps of vector species that could be used to maximize efficiency of surveys and optimize use of economic resources in epidemiology and control.

As for the entire Amazon Region, malaria continues to be a major health public problem in Roraima that presented an Annual Parasitic Index of 85.4 in 2005, the highest in Brazil. Information on anopheline breeding sites is an essential component in malaria control strategies. Aiming to contribute to the limited knowledge on anopheline immature forms in Roraima, collections and breeding site observations were performed in 10 breeding sites around the capital city Boa Vista. Collections were carried out in the rainy and dry season periods between April 2004 and January 2005. Breeding sites comprised natural and artificial water reservoirs. A total of 623 immature forms were collected belonging to Anopheles albitarsis s.l., An.triannulatus s.l., An. nuneztovari/dunhami, An. braziliensis, An. evansae, An. oswaldoi s.l., An. strodei and An. darlingi. An. albitarsis and An. braziliensis were the most frequently found species. Eight larvae of An. darlingi were found in only one breeding site located in the forest. An. triannulatus/An. nuneztovari and An. albitarsis/An. braziliensis were the pairs of species that mostly occurred together. Both pair of species displayed the highest affinity index what might indicate a high compatibility for the same breeding conditions and/or a synergistic co-occurrence. Species diversity index was higher for the dry season.

Toxorhynchites guadeloupensis (Dyar Knab), a poorly known mosquito species, was observed preying upon Aedes aegypti (L.) larvae, in an oviposition trap placed for routine dengue entomological surveillance, during 2003-2004 in the urban area of Boa Vista, Roraima, Brazil. This is the first report for Tx. guadeloupensis using Ae. aegypti oviposition traps as breeding places. This finding may have important consequences in the epidemiology and local dengue control since Ae. aegypti density is a basic variable in dengue prediction. Whether predation of Ae aegypti by Tx. guadeloupensis in the Amazon is of significance, is a question to be examined. Also, larval predation may be a cause for underestimation of the actual Ae aegypti numbers. Together these hypotheses need to be better investigated as they are directly related to dengue epidemiology, to the success of any outbreak prediction and surveillance program.

Parity and age composition for Anopheles darlingi and Anopheles albitarsis in the northern Amazon Basin, Brazil, were investigated. Anopheline ovaries and ovarioles were examined in order to determine whether hourly and seasonal parity status for the vectors An. albitarsis and An. darlingi would vary in two different landscapes (forest and savanna/forest) where malaria is endemic in the northern Amazon Basin. A total of 1,199 anophelines (535 An. darlingi and 664 An. albitarsis) was dissected for parity status, ovariole dilatations, and follicular stages. The total number of nulliparous and parous females for both species varied by time of collection, locality, and season. During the rainy season for the first two h of collection, more nulliparous An. albitarsis and An. darlingi females were collected in the first hour (18:00-19:00), but during the second hour (19:00-20:00) more parous females of both species were captured. During the dry season in Copaíbas, more parous females of An. albitarsis were observed in the first hour while more nulliparous females were observed in the second hour. Nulliparous and parous females of both species for both hours were not significantly different at Road 19 in the dry season. This location was characterized by a forest malaria pattern of transmission with higher numbers of parous females and population stability in the dry season. In Copaíbas, the density and parity of An. darlingi increased during the rainy season, and it could be classified as an alluvial malaria pattern of transmission. For Copaíbas, control measures would be more successful if adopted at the transition from dry to rainy season. Further investigation on longitudinal spatio-temporal change in longevity and survival rates would help us to clarify differences in vector competence for An. darlingi and An. albitarsis and add to the understanding of differences regarding prevailing landscapes in malaria epidemiology in the northern Amazon Basin.

Understanding the different background landscapes in which malaria transmission occurs is fundamental to understanding malaria epidemiology and to designing effective local malaria control programs. Geology, geomorphology, vegetation, climate, land use, and anopheline distribution were used as a basis for an ecological classification of the state of Roraima, Brazil, in the northern Amazon Basin, focused on the natural history of malaria and transmission. We used unsupervised maximum likelihood classification, principal components analysis, and weighted overlay with equal contribution analyses to fine-scale thematic maps that resulted in clustered regions. We used ecological niche modeling techniques to develop a fine-scale picture of malaria vector distributions in the state. Eight ecoregions were identified and malaria-related aspects are discussed based on this classification, including 5 types of dense tropical rain forest and 3 types of savannah. Ecoregions formed by dense tropical rain forest were named as montane (ecoregion I), submontane (II), plateau (III), lowland (IV), and alluvial (V). Ecoregions formed by savannah were divided into steppe (VI, campos de Roraima), savannah (VII, cerrado), and wetland (VIII, campinarana). Such ecoregional mappings are important tools in integrated malaria control programs that aim to identify specific characteristics of malaria transmission, classify transmission risk, and define priority areas and appropriate interventions. For some areas, extension of these approaches to still-finer resolutions will provide an improved picture of malaria transmission patterns.

Objective: To evaluate a new method of deriving the reproductive number for vector-borne diseases from the early epidemic curves for vector-borne diseases with incubations in the vectors and in the hosts., Method: We applied the model to several dengue epidemics in different climatic regions of Brazil: Brasilia, Belém, Fortaleza, Boa Vista., Results: The new method leads to higher estimates of the reproductive number than previous models., Conclusion: At present, Aedes aegypti densities, the meeting of more compatible strains of viruses and mosquitoes, may lead to re-emergence of urban yellow fever epidemics.

Even though Chagas disease is rare in the Brazilian Amazon, the conditions for the establishment of domiciliated cycles prevail in many areas where triatomines are of frequent occurrence. In Roraima, a previous serological and entomological survey in three agricultural settlements showed the existence of all transmission cycle elements, i.e., individuals infected by Trypanosoma cruzi, triatomine species previously found harboring T. cruzi in the broader Amazon region of neighboring countries and, domicile/ peridomicile conditions favorable to triatomine colonization. Triatoma maculata was the most frequent species, found in chicken houses in the peridomicile and sporadically within residences. Aiming to investigate the possibility of T. maculata to possess the potentiality to transmit T. cruzi in the area, bionomic characteristics were studied under laboratory conditions. These were feeding frequency, time for defecation after a blood meal, time elapsed in voluntary fasting pre- and pos-ecdysis, moulting time periods, pre-oviposition and oviposition periods and index of oviposition, incubation period, egg viability, longevity and mortality rate. Results show that the Passarão population of T. maculata should be considered a potential vector of T. cruzi since it shows a capacity to infest artificial ecotopes in the peridomicile, to carry out large number of meals during the nymphal cycle, to have a relatively short developmental cycle capable of producing 2.9 generations/year, to blood source eclecticism, to defecate immediately after the blood meal while still on the host and to the fact that has been previously found naturally infected by T. cruzi.

Chagas disease has been almost entirely eradicated from the arid zones in Central and Northeastern Brazil where rare or no autochthonous cases have been reported. However, in the last 10 years the disease has increasingly been registered in the Amazon Region. Aiming to investigate the possibility of the occurrence of autochthonous cycle of Chagas disease in Roraima, triatomine collections, vectorial susceptibility studies (this one to be shown elsewhere), parasitological and serological analyses were conducted in three agricultural settlement areas (Rorainópolis, Passarão Project and Ilha Community). Blood-donor candidates were also investigated. This is the first epidemiological survey on Chagas disease conducted in agricultural settlements in Roraima. Triatomine species found were Triatoma maculata, Rhodnius pictipes, Rhodnius robustus and Panstrongylus geniculatus. Trypanosoma cruzi detection analyses included xenodiagnosis, indirect immunofluorescence, indirect hemaglutination, ELISA and kinetoplast PCR amplification. Natural triatomine infection was not found in intestinal contents. Twenty-five adult settlers (1.4% out of 1821, all > 15 year-old, 20 migrants) presented anti-T. cruzi antibodies. Two migrant settlers (from Minas Gerais and Maranhão) tested positive for more than two serological tests, besides either being positive for xenodiagnosis or PCR. Results show that Chagas disease is not endemic in the areas studied. However, all elements of the transmission cycle are present, demanding for an adequate and continuous vigilance.

Laboratory-reared Lutzomyia longipalpis (Lutz and Neiva 1912) was tested with extracts of two ichthyotoxic plants, known as timbós, used as fishing poison in the Amazon. Phlebotomines, L. longipalpis, and plants, Antonia ovata and Derris amazonica, were collected in the Raposa-Serra do Sol Indian Reserve, a focus of visceral leishmaniasis in the State of Roraima, Brazil. Extracts were prepared from dried leaves of A. ovata and roots of D. amazonica that were percolated in water, filtered and dried out at 50 degrees C. The solid extract obtained was diluted in water at 150, 200 and 250 mg/ml. The solution was blotted in filter paper placed at the bottom of cylindric glass tubes containing sand flies. For each plant extract and dilution, two series of triplicates with 5 male and 5 female specimens of L. longipalpis were used. Mortality was recorded every 2 h during 72 h of exposure. At 72 h the mortality was as high as 80% for extracts of A. ovata (LD50 = 233 mg/ ml), and 100% for D. amazonica (LD50 = 212 mg/ ml) whereas in the control groups maximum mortality never surpassed 13%. Preliminary assays indicated that A. ovata and D. amazonica displayed significant insecticide effect against L. longipalpis.

The epidemiology of the transmission of malaria parasites varies ecologically. To observe some entomological aspects of the malaria transmission in an urban environment, a longitudinal survey of anopheline fauna was performed in Boa Vista, Roraima, Brazil. A total of 7,263 anophelines was collected in human bait at 13 de Setembro and Caranã districts: Anopheles albitarsis sensu lato (82.8%), An. darlingi (10.3%), An. braziliensis (5.5%), An. peryassui (0.9%) and An. nuneztovari (0.5%). Nightly 12 h collections showed that An. albitarsis was actively biting throughout the night with peak activities at sunset and at midnight. An. darlingi bit during all night and did not demonstrate a defined biting peak. Highest biting indices, entomological inoculation rates and malaria cases were observed seasonally during the rainy season (April-November). Hourly collections showed host seek activity for all mosquitoes peaked during the first hour after sunset. An. darlingi showed the highest plasmodial malaria infection rate followed by An. albitarsis, An. braziliensis and An. nuneztovari (8.5%, 4.6%, 3% and 2.6%, respectively). An. albitarsis was the most frequently collected anopheline, presented the highest biting index and it was the second most frequently collected infected species infected with malaria parasites. An. albitarsis and An. darlingi respectively, are the primary vectors of malaria throughout Boa Vista.

Studies were performed on five Brazilian populations of Lutzomyia longipalpis: Salvaterra (PA), São José do Ribamar (MA), Canindé (CE), Natal (RN) and Gruta da Lapinha, Lagoa Santa (MG). No morphological differences were observed that could distinguish between these populations. Homogeneity tests showed that the allopatric populations display a certain heterogeneity and that the sympatric populations, with different patterns of spots, are homogeneous. The Student-Newman-Keuls test, represented by Euler-Venn diagrams, showed a disjunction between the populations from the north/northeast and the one from Gruta da Lapinha. Genetic distances between the four populations (excluding the Canindé population) were within the range of intrapopulational differences. The Gruta da Lapinha population displayed a heterozygotic deficiency that could be a consequence of high levels of inbreeding due to cryptic habits of living in a small cave. These results do not favor the hypothesis of a L. longipalpis species complex in Brazil, and the species should be considered high polymorphic.

A closed colony of Lutzomyia longipalpis was established with specimens collected in the Raposa - Serra do Sol indian reservoir, one of the main foci of visceral leishmaniasis in the State of Roraima, Brazil. Biological observations were made on four generations of a L. longipalpis colony with emphasis on productivity. Aspects studied were the number of laid and retained eggs, and the number of adults (male and female) per generation. During the four generations the percentage of engorged females that laid eggs varied from 64.2% (third generation-F3) to 90.3% (second generation-F2). The mean number of eggs laid per female varied from 23.6 (F3) to 39. 9 (first generation-F1). The maximum number of eggs laid per female varied from 84 (F3) to 124 (F1). The mean number of retained eggs per female was 12.7 (parental generation-P and F1) to 22.1 (F2). The number of females exceeded the number of males in all generations. However, significant difference for male/female ratio was found only for F3. Fecundity rates were between 42.1 (F3) and 58.3 (F2). From a total of 439 blood-fed females, 355 females laid 12,257 eggs that yield 5,354 adults (2,525 males and 2,829 females) in four generations. F2 presented maximum productivity and fecundity rates.

Samples of the neotropical malaria vector Anopheles darlingi from Bolivia, Brazil, and Venezuela were analyzed to test for differences in mitochondrial haplotype frequencies. With the use of molecular variance components and F-statistics, significant genetic variability of An. darlingi was found apportioned primarily among populations within regions or within populations, with regions defined either as biomes (n = 5) or ecoregions (n = 2). The Mantel analysis resulted in a significant correlation [Prob (r) = 0.009] between genetic and geographic distances, evidence that these populations are genetically isolated by distance. Such isolation could reflect differences in phenotypes for factors affecting vector capacity.

Construction of transgenic Anopheles mosquitoes refractory to Plasmodium requires knowledge of mosquito developmental biology. In order to study Anopheles embryology the removal or, alternatively, the permeabilization of the melanized and sclerotized egg chorion were attempted. The protocol classically used for chorion removal of Drosophila eggs was applied, with partial efficacy, to Anopheles albitarsis, a neotropical malaria vector. Each step was monitored by scanning electron microscopy and the results suggest differences in chorion composition between the two taxa. As an alternative to chorion removal, mosquito eggs were permeabilized with benserazide, an inhibitor of Dopa Decarboxylase, one of the enzymes needed for mosquito eggshell sclerotization. Embryo morphology and viability were not affected by this treatment. Permeabilization of the egg chorion allowed the ultrastructural observation of an internal homogeneous endochorion and an external compound exochorion, the latter consisting of a basal lamellar layer and protruding tubercles.

The development of genetically modified vectors refractory to parasites is seen as a promising strategy in the future control of endemic diseases such as malaria. Nevertheless, knowledge of mosquito embryogenesis, a pre-requisite to the establishment of transgenic individuals, has been presently neglected. We have here studied the eggs from two neotropical malaria vectors. Eggs from Anopheles (Nyssorhynchus) albitarsis and Anopheles (Nyssorhynchus) aquasalis were analyzed by laser scanning microscopy and scanning electron microscopy and compared to those of Drosophila melanogaster. We verified basic conflicting data such as mosquito egg polarity and ultrastructure of eggshell layers. A 180 degrees rotation movement of the mosquito embryo along its longitudinal axis, a phenomenon not conserved among all Diptera, was confirmed. This early event is not taken into account by several present groups, leading to a non-consensual assignment of eggshell dorsal and ventral poles. Since embryo and egg polarities, defined during oogenesis, are the same, we propose to consider the flattened egg side as the dorsal one. The structure of Anopheles eggshell was also examined. Embryos are covered by a smooth endochorion or inner chorion layer. Outside this coat lies the compound exochorion or outer chorion layer, assembled by a thin basal lamellar layer and external tubercles. The terminology related to eggshell layers is discussed.

Three populations of Anopheles darlingi were studied for cuticular hydrocarbons, isoenzymes and patterns of peak biting activity. Differences were found in specimens from Costa Marques, a malaria endemic area; Dourado, a site with a very exophilic population and Juturnaíba, located near the type locality. Twelve hour collections from sunset to sunrise showed that An. darlingi from Costa Marques had a bimodal biting activity profile with a major peak at sunset and a minor peak at sunrise. At Dourado, the pattern was trimodal, with peaks at both morning and evening periods of twilight and near midnight. The Juturnaíba population showed a slight increase in activity near 2000 and 0100 h. Nei's genetic distances, determined by isoenzyme electrophoresis between pairs of populations, were low (D < or = 0.049). Using discriminant analysis for the cuticular hydrocarbons, 92.4% of the specimens from Costa Marques, 91.2% of the specimens from Dourado and 61.3% from Juturnaíba were correctly identified. Cuticular hydrocarbon and isoenzyme results matched very well: the smaller the Nei's distance, the more misidentifications occurred in the jackknife estimator used in the cuticular hydrocarbon analysis. This is the first report of cuticular hydrocarbon analysis in combination with isoenzymes to investigate neotropical anopheline species.