Ranulfo González

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 (phi ST 0.1131, P < 0.001). The variation among individuals within populations (phi ST 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.

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% of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7% 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.

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.

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.

Molecular Taxonomy Provides New Insights into Anopheles Species of the Neotropical Arribalzagia Series

Phylogenetic analysis of partial mitochondrial cytochrome oxidase c subunit I (COI) and nuclear internal transcribed spacer 2 (ITS2) sequences were used to evaluate initial identification and to investigate phylogenetic relationships of seven Anopheles morphospecies of the Arribalzagia Series from Colombia. Phylogenetic trees recovered highly supported clades for An. punctimaculas.s., An. calderoni, An. malefactor s.l., An. neomaculipalpus, An. apicimacula s.l., An. mattogrossensis and An. peryassui. This study provides the first molecular confirmation of An. malefactorfrom Colombia and discovered conflicting patterns of divergence for the molecular markers among specimens from northeast and northern Colombia suggesting the presence of two previously unrecognized Molecular Operational Taxonomic Units (MOTUs). Furthermore, two highly differentiated An. apicimacula MOTUs previously found in Panama were detected. Overall, the combined molecular dataset facilitated the detection of known and new Colombian evolutionary lineages, and constitutes the baseline for future research on their bionomics, ecology and potential role as malaria vectors.

Evidence of Sexual Selection in Neoleucinodes elegantalis (Lepidoptera: Crambidae): Correlation of Female Moth Genitalia and Solanaceae Host Fruit Size

ABSTRACT Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae) is a major pest of fruits in the family Solanaceae in the Western Hemisphere. The objectives of this study were to determine whether life zone or host plant explained morphological variation in females, and if so, if there was evidence of sexual selection driving diversification in this species. We collected larvae feeding on cultivated (Capsicum annuum L., Solanum betaceum Cavanilles, Solanum lycopersicum Lamarck, Solanum melongena L., and Solanum quitoense Lamarck) and wild species (Solanum atropurpureum Schrank, Solanum acerifolium Dunal, Solanum crinitum Lamarck, and Solanum hirtum Vahl) of Solanceae in Colombia. The genitalia traits of 547 reared females were measured and correlations with host plant fruit size were estimated. Six female genitalia morphological characters, apophysis posterioris, apophysis anterioris, ostium bursae, ductus bursae length, corpus bursae, and the seventh abdominal segment were measured. Principal component analysis and cluster analysis classified individuals based on female morphological similarity and clustered them into four main groups according to host plant: 1) S. aceriflolium; 2) S. quitoense, S. lycopersicum, C. annuum and S. hirtum; 3) S. atroporpureum; 4) S. melongena, S. crinitum and S. betaceum. In this unique study, we found that variation in female genitalia size is directly correlated with the size of its host fruit, which suggests a mechanism for reproductive isolation and divergence of the four host races. Ours is one of the first studies that shows female genitalia morphology is correlated with species of host plants and represents a valuable contribution to the study of sexual selection in the evolution of insects.

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.

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.

Anopheles darlingi (Diptera: Culicidae) Rood, 1926: variaciones morfométricas en alas y patas de poblaciones de Colombia

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.

Rediscovery of Antilopsocus nadleri Gurney, 1965 (Psocodea, Troctomorpha, Pachytroctidae) in South America

We found 14 female specimens of Antilopsocus nadleri Gurney, 1965 in Distrito Federal and Minas Gerais, Brazil and in Cauca, Colombia, far from Trinidad and Tobago and Mato Grosso do Sul, Brazil, where the species was previously known. Antilopsocus nadleri had not been collected since 1968.