Morphological and molecular analyses demonstrate identification problems of Anopheles nuneztovari (Diptera: Culicidae) using dichotomous keys
Main Article Content
The Oswaldoi Group, subgenus Nyssorhynchus (Diptera: Culicidae), includes 16 species. Some of these species exhibit high intraspecific morphological variability and interspecific similarity; this can lead to difficulty in the identification of females by using morphological keys, as is the case with Anopheles nuneztovari, considered a primary malaria vector in Colombia. In this study we compared the usefulness of five morphological keys for the identification of A. nuneztovari specimens collected in the locality of Puerto Anchica, Montelibano, Córdoba, Colombia. Three morphometric ratios were analyzed and morphological identification results were confirmed using the PCR-RFLP technique based on ITS2 sequences. The analysis of 41 females using the keys showed that A. nuneztovari specimens presented overlap with other species of the Oswaldoi Group, such as A. rangeli, A. oswaldoi, A. evansae and A. benarrochi. Molecular analysis confirmed that all of the specimens corresponded to A. nuneztovari. Because the use of morphological keys continue to be a strategy of choice for the identification of anophelines, the results suggest that in the case of problematic species, it is convenient to confirm identification using molecular methods developed with the support of the identification of immature stages.
ALAM, M. T.; DAS, M. K.; DEV, V.; ANSARI, M. A.; SHARMA, Y. D. 2007. Identification of two cryptic species in the Anopheles (Cellia) annularis complex using ribosomal DNA PCRRFLP. Parasitology Research 100: 943-948.
ANYANWU, G. I.; DAVIES, D. H.; MOLYNEUX, D. H.; PHILLIPS, A. 1997. Variation in cuticular hydrocarbons among strains of Anopheles (Cellia) stephensi Liston possibly related to prior insecticide exposure. Annals of Tropical Medicine and Parasitology 91: 649-659.
ARRUDA, M.; CARVALHO, M. B.; NUSSENZWEIG, R. S.; MARACIC, M.; FERREIRA A. W.; COCHRANE, A. H. 1986. Potential vectors of malaria and their different susceptibility to Plasmodium falciparum and Plasmodium vivax in northern Brazil identified by inmuno-assay. American Journal of Tropical Medicine and Hygiene 35: 873-881.
BEEBE, N. W.; SAUL, A. 1995. Discrimination of all members of the Anopheles puntulatus complex by polymerase chain reaction-restriction fragment length polymorphism analysis. American Journal of Tropical Medicine and Hygiene 53: 478-481.
BROCHERO, H.; QUIÑONES, M. L. 2008. Retos de la entomología medica para la vigilancia en salud pública en Colombia: reflexión para el caso de malaria. Biomédica 28 (1): 18-24.
CALLE, D. A.; QUIÑONES, M. L.; ERAZO, H. F.; JARAMILLO, N. 2002. Morphometric discrimination of females of five species of Anopheles of the subgenus Nyssorhynchus from southern and northwest Colombia. Memórias do Instituto Oswaldo Cruz 97: 1191-1195.
CALLE, D. A; QUIÑONES, M. L; ERAZO, H. F.; JARAMILLO, N. 2008. Discriminación por morfometría geométrica de once species de Anopheles (Nyssorhynchus) presentes en Colombia. Biomédica 28: 371-385.
CHEN, B.; BUTLIN, R. K.; PEDRO, P. M.; WANG, X. Z.; HARBACH, R. E. 2006. Molecular variation, systematics and distribution of the Anopheles fluviatilis complex in southern Asia. Medical and Veterinary Entomology 20 (1): 33-43.
CIENFUEGOS, A.; CÓRDOBA, L.; GÓMEZ, G.; LUCKHART, S.; CONN, J.; CORREA, M. 2008. Diseño y evaluación de metodologías basadas en PCR-RFLP de ITS2 para la identificación molecular de mosquitos Anopheles spp. (Diptera: Culicidae) de la Costa Pacífica de Colombia. Revista Biomédica 19 (1): 35-44.
COLLINS, F. H.; PASKEWITZ, S. M. 1996. A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species. Insect Molecular Biology 5 (1): 1-9.
COLLINS, F. H.; KAMAU, L.; RANSON, H. A.; VULULE, J. M. 2000. Molecular entomology and prospects for malaria control. Bulletin of the World Health Organization 78 (12): 1412-1423.
DELGADO, N.; RUBIO-PALIS, Y. 1992. Morphometric characterization of the malaria vector Anopheles nuneztovari (Diptera: Culicidae) from western Venezuela. Mosquito Systematics 24: 231-241.
DELGADO, N.; RUBIO-PALIS, Y. 1993. Identification of Anopheles (Nyssorhynchus) (Diptera:Culicidae) occurring in Western Venezuela. Mosquito Systematics 25 (3): 222-230.
DUJARDIN, J. P. 2008. Morphometrics applied to medical entomology. Infection, Genetics and Evolution 8: 875-890.
EDWARDS, K. A.; DOESCHER, L. T.; KANESHIRO, K. Y.; YAMAMOTO, D. 2007. A database of wing diversity in the Hawaiian Drosophila. PLoS One 2 (5): 487.
ELLIOT, R. 1972. The influence of vector behavior on malaria transmission. American Journal of Tropical Medicine and Hygiene 21: 755-763.
FAJARDO, M.; GONZÁLEZ, R.; FIDEL, M.; LÓPEZ, D.; WILKERSON, R.; SALLUM, M. A. 2008. Morphological analysis of three populations of Anopheles (Nyssorhynchus) nuneztovari Gabaldón (Diptera: Culicidae) from Colombia. Memórias do Instituto Oswaldo Cruz 103 (1): 85-92.
FARAN, M. E. 1980. Mosquito studies (Diptera: Culicidae) XXXIV. A revision of the Albimanus section of the subgenus Nyssorhynchus of Anopheles. Contributions of the American Entomological Institute 15: 1-214.
FARAN, M. E.; LINTHICUM, K. J. 1981. A handbook of the Amazonian species of Anopheles (Nyssorhynchus) (Diptera: Culicidae). Mosquito Systematics 13: 1-81.
FERNÁNDEZ, R.; SCHOELER, G.; STANCIL, J. 2004. Presencia de Anopheles (Nyssorhynchus) benarrochi en áreas de selva con transmisión malárica. Revista Peruana de Medicina Experimental y Salud Pública 21 (4): 217-222.
FLORES-MENDOZA, C.; PEYTON, E.; WILKERSON, R.; LOURENÇO-DE-OLIVEIRA, R. 2004. Anopheles (Nyssorhynchus) konderi Galvão and Damasceno: neotype designation and resurrection from synonymy with Anopheles (Nyssohynchus) oswaldoi (Diptera: Culicidae). Proceedings of the Entomological Society of Washington 106: 118-132.
FRITZ, G.; ENGMAN, S.; RODRIGUEZ, R.; WILKERSON, R. 2004. Identification of four vectors of human Plasmodium spp. by multiplex PCR: Anopheles rangeli, An. strodei, An. Triannulatus, and An. Trinkae (Diptera: Culicidae: Nyssorhynchus). Journal of Medical Entomology 41: 1111–1115.
GARROS, C; KOEKEMOER, L. L.; KAMAU, L.; AWOLOLA, T. S.; VAN BORTEL, W.; COETZEE, M.; COOSEMANS, M.; MANGUIN, S. 2004. Restriction fragment length polymorphism method for the identification of major African and Asian malaria vectors within the Anopheles funestus and An. Minimus groups. American Journal of Tropical Medicine and Hygiene 70 (3): 260-265.
GOBERNACIÓN DE CÓRDOBA. 2008. Situación epidemiológica del programa de enfermedades transmitidas por vectores. Secretaría de desarrollo de la salud, Montería, Córdoba, Colombia. 19 p.
GONZÁLEZ, R.; CARREJO, N. 2007. Introducción al estudio taxonómico de Anopheles de Colombia: claves y notas de distribución. Universidad del Valle, Cali, Colombia. 237 p.
GUTIÉRREZ, L. A; NARANJO, N.; JARAMILLO, L. M.; MUSKUS, C. M.; LUCKHART, S.; CONN J. E.; CORREA M. M. 2008. Natural infectivity of Anopheles species from the Pacific and Atlantic Regions of Colombia. Acta Tropica 107: 99-105.
GUTIÉRREZ, L. A; GONZÁLEZ, J. J.; CASTRO, M. I.; GÓMEZ, G. F.; ROSERO, D. A.; LUCKHART, S.; CONN, J. E.; CORREA, M. M. 2009. Species composition and natural infectivity of anthropophilic Anopheles (Diptera: Culicidae) in states of Córdoba and Antioquia, Northwestern Colombia. Memórias do Instituto Oswaldo Cruz 104 (8): 1117-1124.
HAYES, J.; CALDERON, G.; FALCON, R.; ZAMBRANO, V. 1987. Newly incriminated anopheline vectors of human malaria parasites in Junin Department, Peru. Journal of American Mosquito Control Association 33: 418-422.
HRIBAR, L. J. 1994. Geographic variation of male genitalia of Anopheles nuneztovari (Diptera: Culicidae). Mosquito Systematics 26: 132-144.
HRIBAR, L. J. 1995. Costal wing spot variations within and among progeny of single female Anopheles nuneztovari (Diptera: Culicidae). Mosquito Systematics 27: 1-10.
INSTITUTO DE HIDROLOGÍA, METEOROLOGÍA Y ESTUDIOS MEDIOAMBIENTALES (IDEAM). 2008. Disponible en: http:// www.ideam.gov.co. Fecha última revisión: 1 octubre 2008.
INSTITUTO GEOGRÁFICO AGUSTÍN CODAZZI (IGAC). 2002. Atlas de Colombia. Bogotá. Diccionario geográfico de Colombia. Bogotá, Colombia. 342 p.
INSTITUTO NACIONAL DE SALUD –INS. SIVIGILA. 2007. Boletín Epidemiológico. Semana Epidemiológica 52. Subdirección de Vigilancia y Control en Salud Pública. Disponible en: http://www.ins.gov.co/vigilancia/nivel2.php?seccion=34. Fecha última revisión: 2 septiembre 2008.
JIRAKANJANAKIT, N.; LEEMINGSAWAT, S.; DUJARDIN J. P. 2008. The geometry of the wing of Aedes (Stegomyia) aegypti in isofemale lines through successive generations. Infection, Genetics and Evolution 8: 414-421.
LE SUEUR, D.; SHARP, B. L.; APPLETON, C. C. 1992. Dark-scaled areas on adult Anopheles mosquitoes are selectively affected by temperature-related size variation. Medical and Veterinary Entomology 6: 396-398.
LEISNHAM, P. T.; SALA, L. M.; JULIANO, S. A. 2008. Geographic variation in adult survival and reproductive tactics of the mosquito Aedes albopictus. Journal of Medical Entomology 45 (2): 210-221.
LI, C; WILKERSON, R. C. 2005. Identification of Anopheles (Nyssorhynchus) albitarsis complex species (Diptera:Culicidae) using rDNA ITS2-based PCR primers. Memórias do Instituto Oswaldo Cruz 100: 495-500.
MARRELLI, M. T.; SALLUM, M. A.; MARINOTTI, O. 2006. The second internal transcribed spacer of nuclear ribosomal DNA as a tool for Latin American anopheline taxonomy: a critical review. Memórias do Instituto Oswaldo Cruz 101: 817-832.
MATSON, R.; RIOS, C. T.; CHAVEZ, C. B.; GILMAN, R. H.; FLORIN, D.; SIFUENTES, V. L.; GREFFA, R. C.; YORI, P. P.; FERNANDEZ, R.; PORTOCARRERO, D. V.; VINETZ, J. M.; KOSEK, M. 2008. Improved molecular technique for the differentiation of neotropical anopheline species. American Journal of Tropical Medicine and Hygiene 78 (3): 492-498.
MIRABELLO, L.; CONN, J. E. 2008. Population analysis using the nuclear white gene detects Pliocene/Pleistocene lineage divergence within Anopheles nuneztovari in South America. Medical and Veterinary Entomology 22: 109-119.
NORRIS, D. E. 2002. Genetic markers for study of the anopheline vectors of human malaria. International Journal for Parasitology 32 (13): 1607-1615.
OLANO, V.; BROCHERO, H.; SÁENZ, R.; QUIÑONES, M.; MOLINA, J. 2001. Mapas preliminares de la distribución de especies de Anopheles vectores de malaria en Colombia. Biomédica 21: 402-408.
PÉREZ, L.; SUÁREZ, M.; MURCIA, L.; DE LA HOZ, F.; OLANO, V. A.; BROCHERO, H. 1999. La malaria en el Amazonas conocimientos, prácticas, prevalencia de parasitemia y evaluación entomológica en mayo de 1997. Biomédica 19: 93-102.
QUIÑONES, M. L; SUÁREZ, M. F; FLEMING, G. A. 1987. Distribución y bionomía de los anophelinos en la Costa Pacífica de Colombia. Colombia Médica 16: 19-24.
RUBIO-PALIS, Y. 2000. Anopheles (Nyssorhynchus) de Venezuela: taxonomía, bionomía, ecología e importancia médica. Escuela de Malariología y Saneamiento Ambiental “Doctor Arnoldo Gabaldon”. Maracay, Venezuela. 118 p.
RUIZ, F.; QUIÑONES, M. L.; ERAZO, H.; CALLE, D.; ALZATE, J.; LINTON, Y. M. 2005. Molecular differentiation of Anopheles (Nyssorhynchus) benarrochi and An. (N.) oswaldoi from Southern Colombia. Memórias do Instituto Oswaldo Cruz 100: 155-160.
SALLUM, M. A.; BERGO, E. S.; FLORES, D. C.; FORATTINI, O. P. 2002. Systematic studies on Anopheles galvaoi Causey, Deane & Deane from the subgenus Nysssorhynchus blanchard (Diptera: Culicidae). Memórias do Instituto Oswaldo Cruz 97 (8): 1177-1189.
SALLUM, M. A.; MARRELLI, M. T.; NAGAKI S. S.; LAPORTA G. Z.; DOS SANTOS, C. L. 2008. Insight into Anopheles (Nyssorhynchus) (Diptera: Culicidae) species from Brazil. Journal of Medical Entomology 45 (6): 970 -981.
SCARPASSA, V. M.; TADEI, W. P.; SUÁREZ, M. F. 1999. Population structure and genetic divergence in Anopheles nuneztovari< (Diptera: Culicidae) from Brazil and Colombia. The American Journal of Tropical Medicine and Hygiene 60 (6): 1010-1018.
SUÁREZ, M. F.; QUIÑONES, M. L.; ROBAYO, M. A. 1988. Clave para la determinación taxonómica de larvas y adultos hembras de los principales anofelinos de Colombia. Ministerio de Salud, Bogotá, Colombia. 49 p.
TADEI W. P.; DUTARY, T. B.; SANTOS, J.; SCARPASSA, V.; RODRIGUES, I. B.; SILVA, R. M. 1998. Ecologic observations on anopheline vectors of malaria in the Brazilian Amazon. American Journal of Tropical Medicine and Hygiene 59 (2): 325-335.
TRUJILLO, A.; AGUDELO, O.; MOLINA, P.; MORENO, M.; ZAPATA, M; LUCKHART, S.; CORREA, M. 2005. Análisis de PCR-RFLP del espaciador interno transcrito ITS1 y caracterización de las variaciones morfológicas de isofamilias de Anopheles nuneztovari, San Pedro de Urabá, 2004. Laboratorio Actual 38: 81.
VAN BORTEL, W; HARBACH, R. E; TRUNG, H. D; ROELANTS, P; BACKELJAU, T; COOSEMANS, M. 2001. Confirmation of Anopheles varuna in Vietnam previously misidentified and mistargeted as the malaria vector . American Journal of Tropical Medicine and Hygiene 65: 729-732.
WHITMAN, D. W.; ANANTHAKRISHNAN, T. N. 2009. Phenotypic plasticity of insects: mechanisms and consequences. Science Publishers, Enfield, NH, USA. 904 p.
WILKERSON, R. C.; PEYTON, E. L. 1990. Standardized nomenclature for the costal wing spots of the genus Anopheles and other spotted-wing mosquitoes (Diptera: Culicidae). Journal of Medical Entomology 27: 207-224.
WORLD HEALTH ORGANIZATION. 2003. Malaria entomology and vector control. Learner´s guide. Geneva: WHO. 109 p.
ZAPATA, M. A.; CIENFUEGOS, A. V.; QUIRÓS, O. I.; QUIÑONES, M. L.; LUCKHART, S.; CORREA, M. M. 2007. Discrimination of seven Anopheles species from San Pedro de Urabá, Antioquia, Colombia, by polymerase chain reaction-restriction fragment length polymorphism analysis of ITS sequences. American Journal of Tropical Medicine and Hygiene 77 (1): 67-72.
- DORIS A. ROSERO, LINA A. GUTIÉRREZ, ASTRID V. CIENFUEGOS, LUZ M. JARAMILLO, MARGARITA M. CORREA, Optimization of a DNA extraction procedure for anopheline mosquitoes , Revista Colombiana de Entomología: Vol. 36 No. 2 (2010)
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors retain the copyright on their work and are responsible for the ideas expressed in them. Once a manuscript is approved for publication, authors are asked for a publication license for the term of legal protection, for all territories that allows the use, dissemination and disclosure of the same.