Anopheles darlingi Root 1926 (Diptera: Culicidae): variaciones morfométricas en alas y patas de poblaciones de Colombia / Anopheles darlingi (Diptera: Culicidae) Rood 1926: Morphometric variations in wings and legs of populations from Colombia

Resumen Introducción. Las poblaciones naturales de Anopheles darlingi, principal transmisor de la malaria en Colombia, han mostrado plasticidad fenotípica en algunos de sus caracteres diagnósticos. Objetivo. Caracterizar las variaciones morfométricas de los patrones alares y de la pata posterior en poblaciones naturales de An. darlingi recolectados en localidades colombianas donde la malaria es endémica. Materiales y métodos. Se analizaron los patrones de manchas de la vena costal del ala de hembras silvestres recolectadas en los departamentos de Chocó, Guaviare, Meta y Vichada mediante morfometría lineal y geométrica. El segundo tarsómero posterior de las hembras se analizó mediante morfometría lineal. Resultados. Se encontraron 19 patrones de manchas de la vena costal. Los patrones I, con 49 % (n=118/240) y VI, con 28 % (n=66), fueron los más frecuentes. La proporción del área oscura basal (DS-III2 ) y del tarsómero posterior (Ta-III2 ) constituyó una característica sólida de diagnóstico, pues representó el 89 % (n=213/240) del total de especímenes analizados. Se encontraron diferencias significativas en la forma (F=1,65; gl=50; p<0,001) y el tamaño (F=3,37; gl=5; p=0,005) del ala entre las poblaciones de diferentes localidades. El centroide de menor tamaño (2,64 mm) se encontró en las poblaciones de Chocó. Conclusiones. Se registraron 11 patrones nuevos para las manchas de la vena costal y se confirmó la dominancia de los patrones alares I y VI en las poblaciones de An. darlingi de Colombia, así como la relación de DS-III2 y Ta-III2 como un rasgo sólido de diagnóstico para la taxonomía de la especie. Se encontraron diferencias en el tamaño corporal de las poblaciones evaluadas, lo cual reviste importancia para el análisis de aspectos bionómicos de la especie.

Abstract Introduction: Natural populations of Anopheles darlingi, the main malaria vector in Colombia, have shown phenotypic variations in some of their diagnostic characters. Objective: To characterize morphometric variations in wing spot patterns and diagnostic characters of the hind leg of adult females of An. darlingi collected in areas where malaria is endemic. Materials and methods: Patterns of the costal vein spots of wild females of An. darling populations collected in the departments of Chocó, Guaviare, Meta and Vichada were analyzed using linear and morphometric approaches. The second tarsomere of the hind leg of females was characterized by linear morphometric analysis. Results: We found 19 patterns of spots in the costal vein; patterns I (n=118/240, 49%) and VI (n=66, 28%) were the most frequent. The proportion of the basal dark area of hind tarsomere II and the length of hind tarsomere II (DSIII2 /Ta-III2) constituted a robust diagnostic character as it represented 89% (n=213/240) of the total specimens studied. Significant differences were found in the wing shape (F=1.65, df =50, p<0.001) and the wing size (F=3.37, df=5, p=0.005) among populations from different locations. The smallest centroid size (2.64 mm) was found in populations from Chocó. Conclusions. We registered 11 new wing spot patterns in the costal vein and the dominance of the patterns I and VI for populations of An. darlingi from Colombia. We confirmed DSIII2/TaIII2 ratio as a robust diagnostic character for the taxonomy of this species. We found differences between the size and shape of the wings of An. darlingi populations in accordance to their geographical distribution, which constitute important bionomic aspects for this malaria vector.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia) pholidotus (Diptera: Culicidae) from Colombia

Since 1984, Anopheles (Kerteszia) lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Anthropophilic biting behaviour of Anopheles (Kerteszia) neivai Howard, Dyar & Knab associated with Fishermen’s activities in a malaria-endemic area in the Colombian Pacific

On the southwest Pacific Coast of Colombia, a field study was initiated to determine the human-vector association between Anopheles (Kerteszia) neivai and fishermen, including their nearby houses. Mosquitoes were collected over 24-h periods from mangrove swamps, marshlands and fishing vessels in three locations, as well as in and around the houses of fishermen. A total of 6,382 mosquitoes were collected. An. neivai was most abundant in mangroves and fishing canoes (90.8%), while Anopheles albimanus was found indoors (82%) and outdoors (73%). One An. neivai and one An. albimanus collected during fishing activities in canoes were positive for Plasmodium vivax , whereas one female An. neivai collected in a mangrove was positive for P. vivax . In the mangroves and fishing canoes, An. neivai demonstrated biting activity throughout the day, peaking between 06:00 pm-07:00 pm and there were two minor peaks at dusk and dawn. These peaks coincided with fishing activities in the marshlands and mangroves, a situation that places the fishermen at risk of contracting malaria when they are performing their daily activities. It is recommended that protective measures be implemented to reduce the risk that fishermen will contract malaria.

Malaria vector species in Colombia: a review

Here we present a comprehensive review of the literature on the vectorial importance of the major Anopheles malaria vectors in Colombia. We provide basic information on the geographical distribution, altitudinal range, immature habitats, adult behaviour, feeding preferences and anthropophily, endophily and infectivity rates. We additionally review information on the life cycle, longevity and population fluctuation of Colombian Anopheles species. Emphasis was placed on the primary vectors that have been epidemiologically incriminated in malaria transmission: Anopheles darlingi, Anopheles albimanus and Anopheles nuneztovari. The role of a selection of local, regional or secondary vectors (e.g., Anopheles pseudopunctipennis and Anopheles neivai) is also discussed. We highlight the importance of combining biological, morphological and molecular data for the correct taxonomical determination of a given species, particularly for members of the species complexes. We likewise emphasise the importance of studying the bionomics of primary and secondary vectors along with an examination of the local conditions affecting the transmission of malaria. The presence and spread of the major vectors and the emergence of secondary species capable of transmitting human Plasmodia are of great interest. When selecting control measures, the anopheline diversity in the region must be considered. Variation in macroclimate conditions over a species' geographical range must be well understood and targeted to plan effective control measures based on the population dynamics of the local Anopheles species.

Confirmation of Anopheles (Anopheles) calderoni Wilkerson, 1991 (Diptera: Culicidae) in Colombia and Ecuador through molecular and morphological correlation with topotypic material

The morphologically similar taxa Anopheles calderoni, Anopheles punctimacula, Anopheles malefactor and Anopheles guarao are commonly misidentified. Isofamilies collected in Valle de Cauca, Colombia, showed morphological characters most similar to An. calderoni, a species which has never previously been reported in Colombia. Although discontinuity of the postsubcostal pale spots on the costa (C) and first radial (R1) wing veins is purportedly diagnostic for An. calderoni, the degree of overlap of the distal postsubcostal spot on C and R1 were variable in Colombian specimens (0.003-0.024). In addition, in 98.2 percent of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7 percent of specimens examined. Correlation of DNA sequences of the second internal transcribed spacer and mtDNA cytochrome c oxidase subunit I gene (COI) barcodes (658 bp of the COI gene) generated from Colombian progeny material and wild-caught mosquitoes from Ecuador with those from the Peruvian type series of An. calderoni confirmed new country records. DNA barcodes generated for the closely related taxa, An. malefactor and An. punctimacula are also presented for the first time. Examination of museum specimens at the University of the Valle, Colombia, revealed the presence of An. calderoni in inland localities across Colombia and at elevations up to 1113 m.

A population genetics study of Anopheles darlingi (Diptera: Culicidae) from Colombia based on random amplified polymorphic DNA-polymerase chain reaction and amplified fragment lenght polymorphism markers

The genetic variation and population structure of three populations of Anopheles darlingi from Colombia were studied using random amplified polymorphic markers (RAPDs) and amplified fragment length polymorphism markers (AFLPs). Six RAPD primers produced 46 polymorphic fragments, while two AFLP primer combinations produced 197 polymorphic fragments from 71 DNA samples. Both of the evaluated genetic markers showed the presence of gene flow, suggesting that Colombian An. darlingi populations are in panmixia. Average genetic diversity, estimated from observed heterozygosity, was 0.374 (RAPD) and 0.309 (AFLP). RAPD and AFLP markers showed little evidence of geographic separation between eastern and western populations; however, the F ST values showed high gene flow between the two western populations (RAPD: F ST = 0.029; Nm: 8.5; AFLP: F ST = 0.051; Nm: 4.7). According to molecular variance analysis (AMOVA), the genetic distance between populations was significant (RAPD:phiST = 0.084; AFLP:phiST = 0.229, P < 0.001). The F ST distances and AMOVAs using AFLP loci support the differentiation of the Guyana biogeographic province population from those of the Chocó-Magdalena. In this last region, Chocó and Córdoba populations showed the highest genetic flow.

Random amplified polymorphic DNA analysis of Anopheles nuneztovari (Diptera: Culicidae) from Western and Northeastern Colombia

Random amplified polymorphic DNA (RAPD) markers were used to analyze 119 DNA samples of three Colombian Anopheles nuneztovari populations to study genetic variation and structure. Genetic diversity, estimated from heterozygosity, averaged 0.34. Genetic flow was greater between the two populations located in Western Colombia (F ST: 0.035; Nm: 6.8) but lower between these two and the northeastern population (F ST: 0.08; Nm: 2.8). According to molecular variance analysis, the genetic distance between populations was significant (phiST 0.1131, P < 0.001). The variation among individuals within populations (phiST 0.8869, P < 0.001)was also significant, suggesting a greater degree of population subdivision, not considered in this study. Both the parameters evaluated and the genetic flow suggest that Colombian An. nuneztovari populations are co-specific

Man-biting activity of Anopheles (Nyssorhynchus) albimanus and An. (Kerteszia) neivai (Diptera: Culicidae) in the Pacific lowlands of Colombia

The daily man-biting activity of Anopheles (Nyssorhynchus) albimanus and An. (Kerteszia) neivai was determined in four ecologically distinct settlements of the Naya River, Department of Valle, Colombia. Differences were found among the settlements with respect to the mosquito species present, intradomiciliary and extradomiciliary biting activity and population densities.

Ciclo biologico y establecimiento de una colonia de Anopheles pseudopunctipennis the Obald 1901, variedad bifoliata (Diftera: Culicidae) en Cali, Colombia / Biologic cycle and establishment of a colony of Anopheles pseudopunctipennis, the Obald 1901, Bifoliate variety (Diftera: Culicidae> in Cali, Colombia

Anopheles pseudopunctipennis, variedad bifoliata, fue colonizada por cinco generaciones, en condiciones de laboratorio, utilizado copula inducida. Los ejemplares fueron expuestos a un fotoperiodo de 12 horas, 24-29 grados C de temperatura y 72-80% de humedad relativa. Se tomaron medidas de crecimiento y duracion del ciclo biologico. La duracion de los estado inmaduros fue de 12.6 dias manteniendose uniforme el tamano promedio larval en las cinco generaciones, mientras que la longevidad obtenida para las hembras y machos fue de 26.0 y 30.8 dias respectivamente