Charles H. Porter

DNA Barcoding for the Identification of Sand Fly Species (Diptera, Psychodidae, Phlebotominae) in Colombia

Sand flies include a group of insects that are of medical importance and that vary in geographic distribution, ecology, and pathogen transmission. Approximately 163 species of sand flies have been reported in Colombia. Surveillance of the presence of sand fly species and the actualization of species distribution are important for predicting risks for and monitoring the expansion of diseases which sand flies can transmit. Currently, the identification of phlebotomine sand flies is based on morphological characters. However, morphological identification requires considerable skills and taxonomic expertise. In addition, significant morphological similarity between some species, especially among females, may cause difficulties during the identification process. DNA-based approaches have become increasingly useful and promising tools for estimating sand fly diversity and for ensuring the rapid and accurate identification of species. A partial sequence of the mitochondrial cytochrome oxidase gene subunit I (COI) is currently being used to differentiate species in different animal taxa, including insects, and it is referred as a barcoding sequence. The present study explored the utility of the DNA barcode approach for the identification of phlebotomine sand flies in Colombia. We sequenced 700 bp of the COI gene from 36 species collected from different geographic localities. The COI barcode sequence divergence within a single species was <2% in most cases, whereas this divergence ranged from 9% to 26.6% among different species. These results indicated that the barcoding gene correctly discriminated among the previously morphologically identified species with an efficacy of nearly 100%. Analyses of the generated sequences indicated that the observed species groupings were consistent with the morphological identifications. In conclusion, the barcoding gene was useful for species discrimination in sand flies from Colombia.

Mosquitoes of eastern Amazonian Ecuador: biodiversity, bionomics and barcodes

Two snapshot surveys to establish the diversity and ecological preferences of mosquitoes (Diptera: Culicidae) in the terra firme primary rain forest surrounding the Tiputini Biodiversity Station in the UNESCO Yasuní Biosphere Reserve of eastern Amazonian Ecuador were carried out in November 1998 and May 1999. The mosquito fauna of this region is poorly known; the focus of this study was to obtain high quality link-reared specimens that could be used to unequivocally confirm species level diversity through integrated systematic study of all life stages and DNA sequences. A total of 2,284 specimens were preserved; 1,671 specimens were link-reared with associated immature exuviae, all but 108 of which are slide mounted. This study identified 68 unique taxa belonging to 17 genera and 27 subgenera. Of these, 12 are new to science and 37 comprise new country records. DNA barcodes [658-bp of the mtDNA cytochrome c oxidase ( COI ) I gene] are presented for 58 individuals representing 20 species and nine genera. DNA barcoding proved useful in uncovering and confirming new species and we advocate an integrated systematics approach to biodiversity studies in future. Associated bionomics of all species collected are discussed. An updated systematic checklist of the mosquitoes of Ecuador (n = 179) is presented for the first time in 60 years.

Use of DNA barcoding to distinguish the malaria vector Anopheles neivai in Colombia

A reference 535 bp barcode sequence from a fragment of the mitochondrial gene cytochrome oxidase I (COI), acquired from specimens of An. neivai Howard, Dyar & Knab, 1913 from its type locality in Panama, was used as a tool for distinguishing this species from others in the subgenus Kerteszia. Comparisons with corresponding regions of COI between An. neivai and other species in the subgenus (An. bellator Dyar & Knab 1906, An. homunculus Komp 1937, An cruzii Dyar & Knab, 1908 and An. laneanus Corrêa & Cerqueira, 1944) produced K2P genetic distances of 8.3-12.6%, values well above those associated with intraspecific variation. In contrast, genetic distances among 55 specimens from five municipalities in the Colombian Pacific coastal state of Chocó were all within the range of 0-2.5%, with an optimized barcode threshold of 1.3%, the limit for unambiguous differentiation of An. neivai. Among specimens from the Chocó region, 18 haplotypes were detected, two of which were widely distributed over the municipalities sampled. The barcode sequence permits discrimination of An. neivai from sympatric species and indicates genetic variability within the species; aspects key to malaria surveillance and control as well as defining geographic distribution and dispersion patterns.

The Sabethines of Northern Andean Coffee-Growing Regions of Colombia

Abstract Sampling for sabethine mosquitoes occurred intermittently from September 2007 to April 2013 in 17 municipalities, located in 5 departments (divisions) in the northern Andean coffee-growing regions of Colombia. Of the 9 genera within the Sabethini tribe known to occur in the Neotropical region, 6 were encountered including 15 species: Jonhbelkinia ulopus, Limatus durhamii, Sabethes ignotus, Sa. luxodens, Sa. undosus, Shannoniana fluviatilis, Trichoprosopon compressum, Tr. digitatum, Tr. evansae, Tr. pallidiventer s.l., Tr. pallidiventer s.s., Wyeomyia arthrostigma, Wy. oblita, Wy. ulocoma, and Wy. undulata. The species Sa. luxodens and Wy. undulata constitute new records for Colombia. These records broaden the knowledge of this important group that includes some important species related to the arbovirus transmission. Records are from the northern Colombian Andes, a region noted for coffee cultivation and ecotourism.

Wyeomyia (Nunezia) Paucartamboensis, a New Species of Sabethini (Diptera: Culicidae) from the Peruvian Andes with a Diagnosis of the Subgenus Nunezia

Abstract. Wyeomyia (Nunezia)paucartamboensis Porter, new species is described from specimens reared from tank bromeliads growing in humid premontane forest on the eastern slopes of the Peruvian Andes. The description, with relevant illustrations and images, is of the adult male and female, as well as of the pupal and fourth-instar larval stages. In addition, a diagnosis of the subgenus Nunezia is presented with emphasis on differentiation from other subgenera of Wyeomyia.