A Systematic Review on the Viruses of Anopheles Mosquitoes: The Potential Importance for Public Health.

Anopheles mosquitoes are the vectors of Plasmodium, the etiological agent of malaria. In addition, Anopheles funestus 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.

Entomological parameters and population structure at a microgeographic scale of the main Colombian malaria vectors Anopheles albimanus and Anopheles nuneztovari.

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.

Anopheles blood meal sources and entomological indicators related to Plasmodium transmission in malaria endemic areas of Colombia.

Malaria is an important public health problem, caused by Plasmodium parasites which are transmitted by female Anopheles mosquitoes that bite humans to obtain blood. The aim of this work was to identify the blood feeding sources of Anopheles female mosquitoes and calculate their entomological indices in relation to Plasmodium transmission. Mosquitoes were collected in malaria endemic localities of the Bajo Cauca and Pacific regions of Colombia using human landing catch and barrier screens, from 18:00 to 24:00 hr, in 2018-2021. Animal censuses within a radius of ∼250 m were carried out at each sampling site. A total of 2018 Anopheles specimens were collected and the most abundant species were Anopheles (Nys.) darlingi and Anopheles (Nys.) nuneztovari. The highest human biting rate was 77.5 bites per person per night (b/p/n) for An. nuneztovari in Córdoba-Pacific and 17.5 b/p/n for An. darlingi in Villa Grande-Bajo Cauca. Both species were active mainly in indoor unwalled rooms of the houses. Only An. nuneztovari from Córdoba-Pacific was infected with Plasmodium, with an entomological inoculation rate of 91.25 infective bites per year. Detection of blood feeding sources demonstrate that humans were the most common host, however, An. nuneztovari showed a preference for feeding on dogs and An. darlingi on pigs, dogs and Galliformes, rather than humans. These results contribute to entomological surveillance information and provide valuable data that can be used to tailor effective control interventions to minimize human-vector contact in these malaria endemic regions.

Genetic and toxigenic diversity of Bacillus cereus group isolated from powdered foods.

Bacillus cereus is a human pathogenic bacterium that produces emetic and diarrheal foodborne diseases. This study evaluated the genetic and toxigenic diversity in B. cereus group isolates from powdered foods collected in public educational institutions, bakeries and powdered food companies located in Medellín, Colombia. B. cereus was detected in 35 of 305 (11%) powdered food samples and 52 B. cereus were isolated. The presence of ten toxin genes, hblCDAB, nheABC, cytK2, entFM and cesB, was evaluated in the isolates by multiplex PCR. The nheABC operon was found in all isolates (100%), hblCDAB in 22 (42%), hblCDA in 8 (15%) and hblCD in 3 (6%); the cytK2 gene was detected in 32 isolates (62%) and entFM in 32 (62%). Notably, the cesB gene was not detected. According to the presence of toxin genes, fifteen profiles were identified. The predominant toxigenic profile contained all toxin genes but cesB. A large genetic diversity was observed by GTG5 fingerprinting with 46 isolates grouped in seven clusters and the remaining six clustering individually. There was no relationship between toxigenic profiles and genetic clusters, but some genetic clusters seemed to be related to particular powdered food types. In general, the results evidenced high genetic and enterotoxigenic diversity among the B. cereus group isolates.

Enterotoxin Gene Distribution and Genotypes of Bacillus cereussensu lato Isolated from Cassava Starch.

Bacillus cereus is a human pathogenic bacterium found in foods with the potential to cause emesis and diarrhea. This study estimated the presence, toxigenic and genomic diversity of B. cereus s.l. obtained from cassava starch samples collected in bakeries and powdered food companies in Medellín (Colombia). Bacillus cereuss.l. was found in 43 of 75 (57%) cassava starch samples and 98 isolates were obtained. The nheABC, hblCDAB, cytK2, entFM and cesB toxin genes were detected by multiplex PCR and the most frequent operon was nheABC, whereas cesB gene was not found. Twelve toxigenic profiles were determined by the detection of toxin genes, and the most frequent profiles harbored all enterotoxin genes. A broad genomic diversity was detected according to GTG5-PCR fingerprinting results with 76 B. cereus s.l. grouped in sixteen clusters and the 22 isolates clustering separately. No relationship was observed between genomic background and toxigenic profiles. In general, the results showed a high genomic and enterotoxigenic diversity in B. cereus s.l. found in cassava starch. These results should incentive future studies to understand the distribution of B. cereus s.l. isolated on raw materials in comparison with finished products.

Effect of land cover and landscape fragmentation on anopheline mosquito abundance and diversity in an important Colombian malaria endemic region.

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.

Spatial fine-resolution model of malaria risk for the Colombian Pacific region.

OBJECTIVE: To categorise and map, at high resolution, the risk of malaria incidence in the Pacific region, the main malaria-endemic region of Colombia. METHODS: The relationship between the environmental variables Normalized Difference Vegetation Index Normalized Difference Water Index, Topographic Wetness Index, precipitation and temperature with the observed Annual Parasitic Index was evaluated using a generalised linear model. An incidence risk map at a resolution of 1 km2 was constructed and projected to the entire endemic region. Associations of malaria risk categories with both presence records and co-occurrence of the three main malaria vectors were determined. RESULTS: A significant correlation was found for the incidence of malaria with precipitation and Normalized Difference Vegetation Index (R2  = 0.98, P < 0.05), whereas there was no significant correlation with the remaining environmental and topographic variables. Moderate- to high-risk areas were located mainly in central Chocó Department along the San Juan and Atrato rivers and in areas west of the Cauca River and Pacific lowlands of the Andes Mountains. There was a statistically significant relationship for the presence of the two main vectors Anopheles darlingi and Anopheles nuneztovari with the high malaria risk category. Furthermore, malaria risk was directly proportional to the number of co-occurring vector species. CONCLUSIONS: The map obtained provides useful information on the risk of malaria in particular places of the Colombian Pacific region. The data can be used by public entities to optimise the allocation of economic resources for vector control interventions and surveillance.

OBJECTIF: Classifier et cartographier, à haute résolution, le risque d'incidence du paludisme dans la région du Pacifique, la principale région de Colombie où le paludisme est endémique. MÉTHODES: La relation entre les variables environnementales, l'Indice de différence de végétation normalisée, l'Indice différence hydrique normalisée, l'Indice d'humidité topographique, les précipitations et la température avec l'indice parasitique annuel observé a été évaluée à l'aide d'un modèle linéaire généralisé. Une carte des risques d'incidence à une résolution de 1 km2 a été construite et projetée sur l'ensemble de la région endémique. Les associations des catégories de risque de paludisme avec les deux, les registres de présence et la co-occurrence des trois principaux vecteurs du paludisme ont été déterminées. RÉSULTATS: Une corrélation significative a été trouvée pour l'incidence du paludisme avec les précipitations et l'indice de différence de végétation normalisée (R2  = 0,98 ; P < 0,05), alors qu'il n'y avait pas de corrélation significative avec les autres variables environnementales et topographiques. Les zones de risque modéré à élevé étaient principalement situées dans le département central du Chocó, le long des rivières San Juan et Atrato, et dans les zones à l'ouest de la rivière Cauca et des basses régions Pacifiques de la Cordillère des Andes. Il y avait une relation statistiquement significative entre la présence des deux principaux vecteurs Anopheles darlingi et Anopheles nuneztovari et la catégorie à haut risque de paludisme. De plus, le risque de paludisme était directement proportionnel au nombre d'espèces de vecteurs co-occurrents. CONCLUSIONS: La carte obtenue fournit des informations utiles sur le risque de paludisme dans des endroits particuliers de la région Pacifique de la Colombie. Les données peuvent être utilisées par les entités publiques pour optimiser l'allocation des ressources économiques pour les interventions de lutte contre les vecteurs et la surveillance.

Potential distribution of main malaria vector species in the endemic Colombian Pacific region.

OBJECTIVE: To assess the existing fundamental niche, potential distribution and degree of niche overlap for the three main Colombian malaria vectors Anopheles albimanus, Anopheles darlingi and Anopheles nuneztovari in the major malaria endemic Pacific region. METHODS: We used models based on presence records and Normalised Difference Vegetation Index (NDVI) data, created using the maximum entropy algorithm. RESULTS: The three vector species occupied heterogeneous environments, and their NDVI values differed. Anopheles albimanus had the largest niche amplitude and was distributed mainly on coastal areas. Environmentally suitable areas for An. albimanus and An. nuneztovari were the dry forest of inter-Andean Valleys in south-western Colombia, as confirmed for An. albimanus during model validation. There was a slight degree of niche overlap between An. darlingi and An. nuneztovari, and the species co-occurred in humid forests, predominantly in riparian zones of the San Juan and Atrato rivers. CONCLUSION: The information obtained may be used for the implementation of vector control interventions in selected priority areas to reduce malaria risk in this region while optimising resources.

OBJECTIF: Evaluer la niche fondamentale existante, la distribution potentielle et le degré de superposition de niches pour les trois principaux vecteurs colombiens du paludisme: Anopheles albimanus, Anopheles darlingi et Anopheles nuneztovari, dans la principale région endémique du Pacifique pour le paludisme. MÉTHODES: Nous avons utilisé des modèles basés sur des enregistrements de présence et des données de Différence Normalisée d'Indice de Végétation par (NDVI), créés à l'aide de l'algorithme d'entropie maximale. RÉSULTATS: Les trois espèces vectorielles occupaient des environnements hétérogènes et leurs valeurs NDVI différaient. Anopheles albimanus avait la plus grande amplitude de niche et était distribué principalement sur les zones côtières. Les zones convenables environnementalement pour An. albimanus et An. nuneztovari était la forêt sèche des vallées inter-andines du sud-ouest de la Colombie, telles que confirmées pour An. albimanus lors de la validation du modèle. Il y avait un léger degré de superposition de niches entre An. darlingi et An. nuneztovari et les espèces coexistaient dans les forêts humides, principalement dans les zones riveraines des fleuves San Juan et Atrato. CONCLUSION: Les informations obtenues peuvent être utilisées pour la mise en œuvre d'interventions de lutte antivectorielle dans certaines zones prioritaires afin de réduire le risque de paludisme dans cette région tout en optimisant les ressources.

Trans-stadial fate of the gut bacterial microbiota in Anopheles albimanus.

Gut microbiota communities in mosquitoes are influenced among others, by developmental stage. There is evidence that the aquatic environment where larvae feed influences the mosquito gut bacterial community composition with only a subgroup of these bacteria been transmitted trans-stadially to adults. This study evaluated the gut bacterial composition of Anopheles albimanus larvae, emerged and circulating mosquitoes, as well as water from the larval habitat, to elucidate transitions in these bacterial communities and determine the final composition in circulating mosquitoes. A 16S rRNA Illumina sequencing allowed to determine that Proteobacteria was the most abundant phylum in larvae (72.4%), emerged mosquitoes (75%), circulating adults (45.4%) and water from the larval habitat (79.1%). A core microbiome analysis evidenced that Enterobacter, Bacillus and Staphylococcus genera were the core bacterial microbiota (OTUs detected in >90%) in the four groups evaluated. PCoA cluster based on Jaccard and Bray Curtis distances showed two main bacterial clusters, one comprising the emerged and circulating adults, and the other the larvae. The results indicated that the gut microbiota of An. albimanus larvae is composed of bacteria acquired from the larval habitat; then, a rearrangement of the bacterial communities occurs in the trans-stadial passage. However, the higher bacterial richness detected in circulating adults suggests bacterial acquisition from the terrestrial environment where the mosquito feeds. Finally, the trans-stadially passage of some bacteria makes of interest their evaluation as candidates for paratransgenic control.

Genomic and Toxigenic Heterogeneity of Bacillus cereus sensu lato Isolated from Ready-to-Eat Foods and Powdered Milk in Day Care Centers in Colombia.

Bacillus cereus sensu lato (s.l.) is a group of bacteria commonly found in diverse environments, including foods, with potential to cause emesis and diarrhea. In Colombia, it is one of the main foodborne pathogens. The aim of this study was to determine the genomic and toxigenic heterogeneity of B. cereus s.l. isolated from ready-to-eat foods and powdered milk collected in day care centers of Medellin, Colombia. Of 112 B. cereus s.l. isolates obtained, 94% were ß-hemolytic. Toxigenic heterogeneity was established by the presence of nheABC, hblCDAB, cytK2, entFM, and cesB toxigenic genes. The nheABC operon and entFM gene were most frequently detected in the isolates, whereas the cesB gene was not found. According to the toxin genes content, nine toxigenic profiles were identified. A 44% of isolates had profiles with all genes for nonhemolytic enterotoxin, hemolysin BL, and enterotoxin FM production (profiles II and IV). Pulsed-field gel electrophoresis analysis indicated a high genomic heterogeneity among the B. cereus s.l., with 68 isolates grouping into 16 clusters and 33 placed separately in the dendrogram. This study provides useful information on the safety of ready-to-eat foods and powdered milk in day care centers where children, a susceptible population, are exposed and it should incentive for more studies to understand the distribution of different toxin-encoding genes among B. cereus s.l. isolates, enabling detailed risk assessment.

Relationship between land cover and Anophelinae species abundance, composition and diversity in NW Colombia.

Anthropic activities, mainly deforestation, have produced rapid transformation of land cover types in the Urabá region at northwest Colombia. Land cover alterations impact the abundance and composition of the Anophelinae community, affecting malaria transmission dynamics. Therefore, this study used landscape metrics to evaluate the relationship of land cover types with Anophelinae species abundance, composition and diversity in the important malaria endemic Urabá region, in NW Colombia. Orthorectified aerial photographs were used to identify land cover types in four localities of the region. Landscape metrics were obtained and diversity indices were estimated for both, land covers and Anophelinae species collected. The impact of land cover type on the presence and abundance of Anophelinae species was evaluated using a canonical correspondence analysis. Diversity indices showed differences in the Anophelinae community and land covers. The variables with more influence in the Anophelinae community composition were locality, bare soil and the interaction between forest and bare soil covers. The most abundant and dominant species Nyssorhynchus nuneztovari (former Anopheles nuneztovari), related with impacted environments was associated with grass, shrub and bare soil land covers. In conclusion, land covers derived from anthropic activities favored the presence and abundance of the main malaria vectors; but, regardless of differences in landscape, unknown specific factors varying among localities lead to a unique configuration in each site that directly shaped anopheline community composition locally. This information is essential for the development of malaria risk maps and for the design of integrated vector control interventions that include the recognition of the landscape features favoring human-vector contact.

Composition and structure of the culturable gut bacterial communities in Anopheles albimanus from Colombia.

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.

Anopheles species composition and entomological parameters in malaria endemic localities of North West Colombia.

Environmental variations associated with alterations derived from human activities affect mosquito abundance and composition. The objective of this study was to evaluate species abundance, composition, biting behavior and human biting rates for Anopheles specimens collected in localities of an important malaria endemic region in NW Colombia. A total of 2041 specimens belonging to nine species were collected in six malaria endemic localities of the Bajo Cauca region. Anopheles braziliensis was the most abundant species (n = 850, 41.7%) and exhibited the highest human biting rate (64.8 b.p.n), followed by the primary vectors Anopheles nuneztovari (n = 614, 30%) and Anopheles darlingi (n = 368, 18%). Biting activity of the main malaria vectors An. nuneztovari and An. darlingi and the opportunistic species An. braziliensis suggests high biting exposure for humans, mainly from 19:00 h to 23:00 h, and therefore, increased malaria risk at these hours. Regarding mosquito species, Puerto Astilla locality in Nechí municipality exhibited the highest α diversity, but in general, diversity in all localities was low. This information provides the bases for the implementation of targeted and effective vector control interventions directed to reduce human vector-contact.

Factors shaping the gut bacterial community assembly in two main Colombian malaria vectors.

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.

Downscaling incidence risk mapping for a Colombian malaria endemic region.

OBJECTIVE: To map at a fine spatial scale, the risk of malaria incidence for the important endemic region is Urabá-Bajo Cauca and Alto Sinú, NW Colombia, using a new modelling framework based on GIS and remotely sensed environmental data. METHODS: The association between environmental and topographic variables obtained from remote sensors and the annual parasite incidence (API) for the years 2013-2015 was calculated using multiple regression analysis; subsequently, a model was constructed to estimate the API and to project it to the entire endemic region in order to design the risk map. The model was validated by relating the obtained API values with the presence of the three main Colombian malaria vectors, Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. RESULTS: Temperature and Normalized Difference Water Index (NDWI) showed a significant correlation with the observed API. The risk map of malaria incidence showed that the zones at higher risk in the Urabá-Bajo Cauca and Alto Sinú region were located south-east of the region, while the northern area presented the lowest malaria risk. A method was generated to estimate the API for small urban centres, instead of the used reports at the municipality level. CONCLUSIONS: These results provide evidence of the utility of risk maps to identify environmentally vulnerable areas at a fine spatial resolution in the Urabá-Bajo Cauca and Alto Sinú region. This information contributes to the implementation of vector control interventions at the microgeographic scale at areas of high malaria risk.

Discrimination of Anopheles species of the Arribalzagia Series in Colombia using a multilocus approach.

The Arribalzagia Series of the Anopheles Subgenus comprises morphologically similar species or members of species complexes which makes correct species identification difficult. Therefore, the aim of this work was to discriminate the morphospecies of the Arribalzagia Series present in Colombia using a multilocus approach based on ITS2, COI and CAD sequences. Specimens of the Arribalzagia Series collected at 32 localities in nine departments were allocated to seven species. Individual and concatenated Bayesian analyses showed high support for each of the species and reinforced the previous report of the Apicimacula species Complex with distribution in the Pacific Coast and northwestern Colombia. In addition, a new molecular operational taxonomic unit-MOTU was identified, herein denominated near Anopheles peryassui, providing support for the existence of a Peryassui species Complex. Further, the CAD gene, just recently used for Anopheles taxonomy and phylogeny, demonstrated its power in resolving phylogenetic relationships among species of the Arribalzagia Series. The divergence times for these species correspond to the early Pliocene and the Miocene. Considering the epidemiological importance of some species of the Series and their co-occurrence in malaria endemic regions of Colombia, their discrimination constitutes an important step for vector incrimination and control in the country.

Mosquito (Diptera: Culicidae) grouping based on larval habitat characteristics in high mountain ecosystems of Antioquia, Colombia.

Information about mosquito ecology in the high mountain ecosystems of the Neotropical region is sparse. In general, few genera and species have been reported in these ecosystems and there is no information available on habitats and the mosquitoes occupying them. In the present study, specimens collected from NW Colombia in HME were grouped using larval habitat data via an Operational Taxonomic Unit (OTU) determination. A total of 719 mosquitoes was analyzed belonging to 44 OTUs. The analysis considered habitat features and clustered the specimens into six groups from A-F. Five of these included species from different genera, suggesting common habitat requirements. Group E with four genera, seven subgenera, and six species occupied the highest areas (above 3,000 m), whereas three groups (B, D, F) were detected at lower altitudes (1,960-2,002 m). Bromeliads were the most common larval habitat, with 47% (335/719) of the specimens; five genera, six subgenera, and eight species were identified and classified into 66% (29/44) of the OTUs. This work showed some similarities to the habitat requirements and provides a grouping system that constitutes an important baseline for the classification of mosquito fauna from high mountain ecosystems according to altitude and larval habitat.

Subtyping of Clostridium difficile PCR ribotypes 591, 106 and 002, the dominant strain types circulating in Medellin, Colombia.

We aimed to achieve a higher typing resolution within the three dominant Clostridium difficile ribotypes (591,106 and 002) circulating in Colombia. A total of 50 C. difficile isolates we had previously typed by PCR-ribotyping, representing the major three ribotypes circulating in Colombia, were analyzed. Twenty-seven isolates of ribotype 591, 12 of ribotype 106 and 11 of ribotype 002 were subtyped by multiple locus variable-number tandem-repeat analysis (MLVA). The presence of the PaLoc genes (tcdA/tcdB), toxin production in culture and antimicrobial susceptibility were also determined. From the total C. difficile ribotypes analyzed, 20 isolates (74%) of ribotype 591, nine (75%) of ribotype 106 and five (45.5%) of ribotype 002 were recovered from patients with Clostridium difficile infection (CDI). MLVA allowed us to recognize four and two different clonal complexes for ribotypes 591 and 002, respectively, having a summed tandem-repeat difference (STRD) <2, whereas none of the ribotype 106 isolates were grouped in a cluster or clonal complex having a STRD >10. Six ribotype 591 and three ribotype 002 isolates belonging to a defined clonal complex were isolated on the same week in two different hospitals. All ribotypes harbored either tcdA+/tcdB+ or tcdA-/tcdB+ PaLoc genes. Moreover, 94% of the isolates were positive for toxin in culture. All isolates were susceptible to vancomycin and metronidazole, while 75% to 100% of the isolates were resistant to clindamycin, and less than 14.8% of ribotype 591 isolates were resistant to moxifloxacina. No significant differences were found among ribotypes with respect to demographic and clinical patients' data; however, our results demonstrated a high molecular heterogeneity of C. difficile strains circulating in Colombia.

[Abundance, composition and natural infection of Anopheles mosquitoes from two malaria-endemic regions of Colombia].

INTRODUCTION: In Colombia there are three Anopheles species implicated in malaria transmission as primary vectors; however, the local role of some Anopheles species must still be defined. OBJECTIVE: To determine the abundance, composition and natural infection rates for Anopheles mosquitoes with Plasmodium spp. in two malaria-endemic regions of Colombia. MATERIALS AND METHODS: Anopheles mosquitoes were collected using the human-landing catches and while resting in livestock corrals in nine localities of two malaria-endemic regions of Colombia. Mosquitoes were morphologically identified and confirmed by PCR-RFLP-ITS2. Identified mosquitoes were processed and tested for Plasmodium parasite infection by ELISA and ssrRNA-based nested PCR. RESULTS: We collected 1,963 Anopheles mosquitoes corresponding to nine species. The most abundant species were Anopheles nuneztovari (53.5%) and A. darlingi (34.5%), followed by A. triannulatus s.l. (6%), and other species (≈5.9%). Three species were naturally infected with Plasmodium spp.: A. darlingi, A. nuneztovari and A. triannulatus s.l. CONCLUSIONS: Natural infection of A. darlingi and A. nuneztovari indicate that these malaria vectors continue to be effective carriers of Plasmodium in the localities under study in Valle del Cauca and Chocó. Additionally, the infected A. triannulatus s.l. collected in livestock corrals in the locality of the department of Córdoba suggests the need for further studies to define the epidemiological importance of this species given its abundance and opportunistic anthropophilic behavior.

Genetic structure and phenotypic variation of Anopheles darlingi in northwest Colombia.

This study evaluated the influence of environmental heterogeneity on Anopheles darlingi genetic and morphometric traits at a microgeographic level. Specimens of An. darlingi collected from multiple municipalities in the Colombian malaria endemic region Urabá-Bajo Cauca and Alto Sinú (UCS) were analyzed using 13 microsatellite loci. Spatial genetic structure, population variation and wing geometric morphometric analyses were performed. Microsatellite results showed low genetic differentiation and high gene flow among populations; four highly admixed subpopulations were detected with no particular association to the municipalities. Wing geometric morphometrics analysis showed a subtle but significant difference in wing shape for El Bagre vs. Mutatá populations, possibly influenced by geographical distance. Discrimination among populations in the morphospace showed a slight separation of the Tierralta population. There was no significant correlation between the genetic and geographic or genetic and environmental distances. We hypothesize that environmental heterogeneity in the UCS region does not reach a threshold to affect population structure of An. darlingi. Another possibility is that microsatellites are not sensitive enough to detect existing structure. It remains to be determined which local factors govern phenotypic variation among these populations and how, or whether these may affect mosquito biology and transmission capacity.