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Anopheles darlingi is a main malaria vector in the neotropical region, but its viral component is not well studied, especially in the neotropics. This work aimed to analyze the virome in Anopheles darlingi from malaria endemic regions of Colombia. Specimens were collected from the Bajo Cauca, Chocoan Pacific and northwestern Amazonas regions and analyzed using an RNA-Seq approach. Results revealed a variety of RNA viral sequences with homology to those of Insect-Specific Viruses belonging to Rhabdoviridae , Partitiviridae , Metaviridae , Tymoviridae , Phasmaviridae , Totiviridae , Ortervirales and Riboviria . Despite geographical and ecological differences among regions, the An. darlingi viral composition remains consistent in different areas, with a core group of viral operational taxonomic units-vOTUs shared by the populations. Furthermore, diversity analysis uncovered greater dissimilarities in viral sequence among mosquitoes from geographically distant regions, particularly evident between populations located at both sides of the Andes Mountain range. This study provides the first characterization of the metavirome in An. darlingi from Colombia and lays the foundation for future research on the complex interactions among viruses, hosts, and microbiota; it also opens a new line of investigation on the viruses in Anopheles populations of Colombia.

Anopheles species composition, abundance, and Plasmodium natural infection are important aspects to assess malaria transmission risk. In this study, these aspects were evaluated in a large number of localities in the most important malaria-endemic regions of Colombia. Mosquitoes were collected from 2012 to 2015 in 55 localities of northwestern and western Colombia. Anopheles species composition, abundance, and Plasmodium infection were estimated. A total of 13,218 Anopheles specimens were evaluated. The highest species richness was detected in the northwest, where the main vectors, An. albimanus (27.8%) and An. nuneztovari (26.7%), were the most abundant species. In the west, An. nuneztovari predominated (51.6%), followed by An. darlingi (29.2%). Six species were infected with Plasmodium, An. darlingi, An. nuneztovari, An. albimanus, An. calderoni, An. triannulatus, and An. braziliensis. Results showed that in these localities the main Colombian vectors are widely spread, which suggests a high malaria risk. Notably, this study is the first to report An. braziliensis from Colombia as being naturally infected with Plasmodium. Infection results for species that are suspected local vectors indicate the importance of conducting further studies to assess their epidemiological importance. This information provides the basis for the application of directed vector control strategies that are species-specific.

Population subdivision among several neotropical malaria vectors has been widely evaluated; however, few studies have analyzed population variation at a microgeographic scale, wherein local environmental variables may lead to population differentiation. The aim of the present study was to evaluate the genetic and geometric morphometric structure of Anopheles nuneztovari and Anopheles albimanus in endemic localities of northwestern Colombia. Genetic and phenetic structures were evaluated using microsatellites markers and wing geometric morphometrics, respectively. In addition, entomological indices of importance in transmission were calculated. Results showed that the main biting peaks of Anopheles nuneztovari were between 20:00 and 22:00, whereas Anopheles albimanus exhibited more variation in biting times among localities. Infection in An . nuneztovari by Plasmodium spp. (IR: 4.35%) and the annual entomological inoculation rate (30.31), indicated high vector exposure and local transmission risk. We did not detect Plasmodium -infected An . albimanus in this study. In general, low genetic and phenetic subdivision among the populations of both vectors was detected using a combination of phenotypic, genetic and environmental data. The results indicated high regional gene flow, although local environmental characteristics may be influencing the wing conformation differentiation and behavioral variation observed in An . albimanus . Furthermore, the population subdivision detected by microsatellite markers for both species by Bayesian genetic analysis provides a more accurate picture of the current genetic structure in comparison to previous studies. Finally, the biting behavior variation observed for both vectors among localities suggests the need for continuous malaria vector surveys covering the endemic region to implement the most effective integrated local control interventions.

The understanding of factors affecting the gut bacterial communities in malaria vectors is essential for the design of vector control interventions, such as those based on a paratransgenic approach. One of the requirements of this method is the availability of bacteria from the mosquito susceptible to culture. Thus, the aim of this study was to evaluate the composition and structure of the culturable gut bacterial communities in field mosquitoes Anopheles albimanus from Colombia, in addition to generate a bacterial collection to further analyze microbial functional activity. Gut bacteria were isolated from An. albimanus larvae and adult mosquitoes collected in localities of the Atlantic and Pacific Coasts. The bacterial isolates were grouped in 28 morphospecies that corresponded to three phyla, three classes, nine families and 14 genera. The larvae guts from San Antero (Atlantic Coast) and Buenaventura (Pacific Coast) shared the genera Bacillus and Lysinibacillus and in adults, Bacillus and Bacillus cereus Group were registered in both localities. Gut bacterial richness was higher in adults from the Pacific with respect to the Atlantic Coast, while larval richness was similar in samples of both coasts. The Shannon index indicated uniformity in morphospecies abundances in both localities. Finally, the characterization of morphospecies from the gut of Anopheles albimanus mosquitoes from Colombia by culture-dependent methods complemented with 16S rRNA gene sequencing allowed the definition, at a finer resolution, of the composition and structure of these microbial communities. In addition, the obtained bacterial culture collection will allow further evaluation of the microorganisms for their potential as biocontrol agents.

Several species of Anopheles mosquitoes (Arthropoda, Insecta, Diptera, Culicidae) are important in public health due to their role in malaria transmission. Of the more than 500 Anopheles species worldwide, 47 have been reported in Colombia, but only nine are known to transmit malaria. Taxonomic classification of these mosquitoes is complicated by the existence of species complexes and groups of closely related species that are difficult to distinguish based on morphological characteristics. However, molecular techniques have contributed to resolving taxonomic uncertainties, definition of molecular variants and facilitated the correction of erroneous taxonomic assignments. This study aimed to update the list of Anopheles species reported for Colombia. A species checklist was compiled by reviewing catalogs, publications, databases, and unpublished data. Only formally characterized species were included, along with their geographic range and ecological distribution. The final list includes 44 formally characterized Anopheles species belonging to five subgenera. The Nyssorhynchus subgenus constituted the largest group with 17 species and the widest distribution, occurring in 18 ecoregions. The Anopheles subgenus was the second largest group with 16 species and occurrences in 16 ecoregions. Sixty-six new presence records were added to the checklist. The updated Anopheles checklist, encompassing presence records and ecological distributions, enhances our understanding of Anopheles mosquito biodiversity. Furthermore, it contributes to improved public health by providing a foundation for targeted vector control interventions.

Landscape structure influences the distribution and abundance of anopheline mosquitoes and has an indirect impact on malaria transmission. This work aimed to determine the effect of land cover and landscape fragmentation on anopheline mosquito abundance and diversity in an important Colombian malaria endemic area, the Bajo Cauca region. Diversity indices were calculated for Anopheles mosquitoes collected in various localities of the region. Land cover types were characterized using orthorectified aerial photographs to estimate landscape metrics. The relationship between landscape fragmentation and species diversity was evaluated by regression analysis. The correlation between species abundance and land cover types was determined using canonical correspondence analyses. Results showed a statistically significant tendency for a lower diversity of the Anopheles community in landscapes with higher patch number, patch density and effective mesh size. For most species, there was evidence of a significant relationship between species abundance and land covers modified by anthropic activities which generate forest loss. These results indicate that activities that modify the landscape structure and land cover composition generate changes that affect the spatial distribution and composition of epidemiologically-important Anopheles species, which may impact malaria distribution in a region. This information is useful to guide control interventions that promote unfavorable landscapes for malaria vector propagation.

Rapid transformation of natural ecosystems changes ecological conditions for important human disease vector species; therefore, an essential task is to identify and understand the variables that shape distributions of these species to optimize efforts toward control and mitigation. Ecological niche modeling was used to estimate the potential distribution and to assess hypotheses of niche similarity among the three main malaria vector species in northern Colombia: Anopheles nuneztovari, An. albimanus, and An. darlingi. Georeferenced point collection data and remotely sensed, fine-resolution satellite imagery were integrated across the Urabá -Bajo Cauca-Alto Sinú malaria endemic area using a maximum entropy algorithm. Results showed that An. nuneztovari has the widest geographic distribution, occupying almost the entire study region; this niche breadth is probably related to the ability of this species to colonize both, natural and disturbed environments. The model for An. darlingi showed that most suitable localities for this species in Bajo Cauca were along the Cauca and Nechí river. The riparian ecosystems in this region and the potential for rapid adaptation by this species to novel environments, may favor the establishment of populations of this species. Apparently, the three main Colombian Anopheles vector species in this endemic area do not occupy environments either with high seasonality, or with low seasonality and high NDVI values. Estimated overlap in geographic space between An. nuneztovari and An. albimanus indicated broad spatial and environmental similarity between these species. An. nuneztovari has a broader niche and potential distribution. Dispersal ability of these species and their ability to occupy diverse environmental situations may facilitate sympatry across many environmental and geographic contexts. These model results may be useful for the design and implementation of malaria species-specific vector control interventions optimized for this important malaria region.

Background The understanding of the roles of gut bacteria in the fitness and vectorial capacity of mosquitoes that transmit malaria, is improving; however, the factors shaping the composition and structure of such bacterial communities remain elusive. In this study, a high-throughput 16S rRNA gene sequencing was conducted to understand the effect of developmental stage, feeding status, species, and geography on the composition of the gut bacterial microbiota of two main Colombian malaria vectors, Anopheles nuneztovari and Anopheles darlingi . Results The results revealed that mosquito developmental stage, followed by geographical location, are more important determinants of the gut bacterial composition than mosquito species or adult feeding status. Further, they showed that mosquito gut is a major filter for environmental bacteria colonization. Conclusions The sampling design and analytical approach of this study allowed to untangle the influence of factors that are simultaneously shaping the microbiota composition of two Latin-American malaria vectors, essential aspect for the design of vector biocontrol strategies.

'Anopheles' mosquitoes are the vectors of 'Plasmodium', the etiological agent of malaria. In addition, 'Anopheles funestu' and 'Anopheles gambiae' are the main vectors of the O'nyong-nyong virus. However, research on the viruses carried by Anopheles is scarce; thus, the possible transmission of viruses by 'Anopheles' is still unexplored. This systematic review was carried out to identify studies that report viruses in natural populations of Anopheles or virus infection and transmission in laboratory-reared mosquitoes. The databases reviewed were EBSCO-Host, Google Scholar, Science Direct, Scopus and PubMed. After the identification and screening of candidate articles, a total of 203 original studies were included that reported on a variety of viruses detected in 'Anopheles' natural populations. In total, 161 viruses in 54 species from 41 countries worldwide were registered. In laboratory studies, 28 viruses in 15 'Anopheles' species were evaluated for mosquito viral transmission capacity or viral infection. The viruses reported in 'Anopheles' encompassed 25 viral families and included arboviruses, probable arboviruses and Insect-Specific Viruses (ISVs). Insights after performing this review include the need for (1) a better understanding of Anopheles-viral interactions, (2) characterizing the 'Anopheles' virome-considering the public health importance of the viruses potentially transmitted by 'Anopheles' and the significance of finding viruses with biological control activity-and (3) performing virological surveillance in natural populations of 'Anopheles', especially in the current context of environmental modifications that May potentiate the expansion of the 'Anopheles' species distribution.

The Colombian Pacific region is second nationally in number of malaria cases reported. This zone presents great ecological heterogeneity and Anopheles species diversity. However, little is known about the current spatial and temporal distribution of vector species. This study, conducted in three ecologically different localities of the Pacific region, aimed to evaluate the composition and distribution of Anopheles species and characterize transmission intensity. A total of 4,016 Anopheles mosquitoes were collected representing seven species. The composition and dominant species differed in each locality. Three species were infected with malaria parasites: Anopheles darlingi and An. calderoni were infected with Plasmodium falciparum and An. nuneztovari with Plasmodium vivax VK210 and VK247. Annual EIRs varied from 3.5-7.2 infective bites per year. These results confirm the importance of the primary vector An. nuneztovari in areas disturbed by human interventions, of An. darlingi in deforested margins of humid tropical rainforest and An. albimanus and the suspected vector An. calderoni in areas impacted by urbanization and large-scale palm oil agriculture close to the coast. This constitutes the first report in the Colombia Pacific region of naturally infected An. darlingi, and in Colombia of naturally infected An. calderoni. Further studies should evaluate the epidemiological importance of An. calderoni in the Pacific region.