Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia

ABSTRACT: Anopheles (Nyssorhynchus) benarrochi, An. (N.) oswaldoi, and An. (N.) rangeli are the most common anthropophilic mosquitoes in the southern Colombian state of Putumayo. Adult females are most commonly collected in epidemiological studies, and this stage poses significant problems for correct identification, due to overlapping inter-specific morphological characters. Although An. rangeli is easy to identify, the morphological variant of An. benarrochi found in the region and An. oswaldoi are not always easy to separate. Herein we provide a rapid molecular method to distinguish these two species in Southern Colombia. Sequence data for the second internal transcribed spacer (ITS2) region of rDNA was generated for link-reared progeny of An. benarrochi and An. oswaldoi, that had been identified using all life stages. ITS2 sequences were 540 bp in length in An. benarrochi (n = 9) and 531 bp in An. oswaldoi (n = 7). Sequences showed no intra-specific variation and ungapped inter-specific sequence divergence was 6.4%. Species diagnostic banding patterns were recovered following digestion of the ITS2 amplicons with the enzyme Hae III as follows: An. benarrochi (365, 137, and 38 bp) and An. oswaldoi (493 and 38 bp). This polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay provides rapid, accurate, and inexpensive species diagnosis of adult females. This will benefit future epidemiological studies and, as PCR amplification can be achieved using a single mosquito leg, the remaining specimen can be either retained as a morphological voucher or further used in vector incrimination studies. That An. benarrochi comprises a complex of at least two species across Latin America is discussed.

• Autores: Quiñones Pinzón, Martha Lucía; Calle Londoño, David Alonso; Erazo, Holmes Francisco; Alzate Restrepo, Juan Fernando; Ruiz López, Jhon Fredy; Linton, Yvonne Marie
• Tipo de recurso: Article of investigation
• Fecha de publicación: 2005
• Institución: Universidad de Antioquia
• Repositorio: Repositorio UdeA
• Idioma: eng
• OAI Identifier: oai:bibliotecadigital.udea.edu.co:10495/39016
• Acceso en línea: https://hdl.handle.net/10495/39016
• Palabra clave: Anopheles; Secuencia de Bases; Base Sequence; Colombia; ADN Espaciador Ribosómico - genética; DNA, Ribosomal Spacer - genetics; Genes de Insecto - genética; Genes, Insect - genetics; Datos de Secuencia Molecular; Molecular Sequence Data; Reacción en Cadena de la Polimerasa; Polymerase Chain Reaction; Polimorfismo de Longitud del Fragmento de Restricción; Polymorphism, Restriction Fragment Length; Análisis de Secuencia de ADN; Sequence Analysis, DNA; https://id.nlm.nih.gov/mesh/D000852; https://id.nlm.nih.gov/mesh/D001483; https://id.nlm.nih.gov/mesh/D003105; https://id.nlm.nih.gov/mesh/D021903; https://id.nlm.nih.gov/mesh/D017344; https://id.nlm.nih.gov/mesh/D008969; https://id.nlm.nih.gov/mesh/D016133; https://id.nlm.nih.gov/mesh/D012150; https://id.nlm.nih.gov/mesh/D017422
• Rights: openAccess
• License: http://creativecommons.org/licenses/by/2.5/co/