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.

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.

Se determinaron las diferencias en desarrollo, sobrevivencia y mortalidad de tres poblaciones del mosquito Culex quinquefasciatus procedente de tres pisos térmicos del departamento de Cundinamarca: Mosquera (2516 msnm, 15°C), Fusagasugá (1728 msnm, 22°C) y Girardot (269 msnm, 28°C). Las larvas y adultos de cada población se mantuvieron en el laboratorio en condiciones similares a los sitios de origen. Se registraron diariamente la supervivencia y mortalidad en cada colonia. Para el análisis estadístico se utilizó la prueba de Kruskal Wallis con la modificación de Bonferroni. Los tiempos de desarrollo por sitio fueron así: Mosquera 15,05 días, Fusagasugá 10,62 días y Girardot 7,72 días. Se encontraron diferencias significativas solamente entre Mosquera y Girardot, pero al analizar el tiempo de desarrollo solo de las larvas hubo diferencias significativas entre las tres poblaciones. Las diferencias intrapoblacionales en las tres cohortes no fueron significativas. La supervivencia y la mortalidad de los estadios pre-reproductivos no mostraron diferencias significativas en las tres cohortes y la supervivencia muestra una curva tipo I. La expectativa de vida de un huevo recién ovipositado procedente de Girardot fue 26,06 días, de Fusagasugá fue de 24,12 días y de Mosquera 40,77 días; las diferencias entre ellos no fueron significativas. Estos datos muestran que Culex quinquefasciatus se ha ido adaptando a zonas más altas con temperaturas menores sin detrimento en los parámetros de la tabla de vida.

The differences in development time, survival, mortality and life expectancy were determined for three populations of the mosquito Culex quinquefasciatus from three temperature zones in the Cundinamarca department of Colombia: Mosquera (2516 m, 15°C), Fusagasugá (1728 m, 22°C) and Girardot (269 m, 33°C). The larvae and adults from each population were kept in laboratory conditions similar to those of their place of origin. Survival and mortality for each colony were recorded daily. For the statistical analysis, the Kruskal Wallis test was used with Bonferroni´s modification. Development times by site were as follows: Mosquera 15.05 days, Fusagasugá 10.62 days and Girardot 7.72 days. Significant differences were only found between Mosquera and Girardot, but by analysis of the development time for larvae separately, were significant differences among all three populations. Intrapopulation differences in the three cohorts were not significant. Survival and mortality of the pre-reproductive stages showed no significant differences in the three cohorts and survival showed a type-1 curve. The life expectancy for a recently oviposited egg from Girardot was 26.06 days, from Fusagasugá it was 24.12 days and from Mosquera it was 40.77 days. These data show that Culex quinquefasciatus has been adapting to higher elevation zones with lower temperatures without detriment to life table parameters.

Objetivo Evaluar el desempeño de las pruebas empleadas en Colombia para el diagnóstico de la leishmaniasis visceral canina y adaptar una técnica de Western blot empleando animales experimental y naturalmente infectados. Metodología Se obtuvieron sueros de 10 perros infectados experimentalmente con L. infantum, 5 perros infectados naturalmente, 16 perros sanos, 26 de reacción cruzada (infectados con Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi, Leishmania (Viannia) spp.), 40 de zonas no endémicas y 150 de zona endémica. Todos fueron evaluados mediante las pruebas de inmunofluorescencia indirecta (IFI), ELISA y Western blot (WB). Resultados Se encontró que IFI tuvo el mayor porcentaje de positividad en los perros infectados (73 %) mientras que el menor porcentaje de falsos positivos se obtuvo por WB (2,5 %). La prueba de ELISA fue la menos eficiente. Fueron reconocidas 24 fracciones antigénicas, las bandas de 29, 34, 50, 69, 75, 86, 99 y 123 kDa fueron responsables de reacciones inespecíficas en los sueros de perros sanos, de zona no endémica y de reacción cruzada. Las bandas por debajo de 29 kDa mostraron ser potencialmente diagnósticas, especialmente la fracción de 13 kDa. Conclusiones Los métodos directos y serológicos pueden subdiagnosticar la infección por Leishmania, solamente un constructo que combine tanto pruebas directas como indirectas sería la forma más eficiente de diagnóstico.

Objective Evaluating canine visceral leishmaniasis diagnostic test performance in Colombia and adapting the Western blot test in naturally and experimentally infected dogs. Methods Sera were obtained from 10 experimentally L. Infantum-infected dogs, 5 naturally infected dogs, 16 healthy dogs, 26 Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi and Leishmania (Viannia) spp infected dogs, 40 dogs from non-endemic areas and 150 from endemic areas. Sera were tested for L. infantum infection using immunofluorescent antibody (IFAT), ELISA and Western blot (WB) tests. Results Positives results were obtained for 73 % of known infected dogs by the IFAT test and false positives were obtained for 2.5 % of non-infected dogs using WB. ELISA was not efficient for diagnosis. 24 antigenic fractions were recognised in tested sera using WB; however, 29, 34, 50, 69, 75, 86, 99 and 123 kDa bands were recognised in sera from dogs from non-endemic areas, healthy dogs and Trypanosoma cruzi, Erhlichia canis, Dirofilaria immitis and Babesia canis infected dogs. The 13 kDa fraction proved potentially useful for diagnosing canine visceral leishmaniasis. ConclusionsThe separate use of parasitological and serological test could lead to misdiagnosis of Leishmania infection; using both kinds of technique simultaneously is thus highly recommended.

Objective Evaluating canine visceral leishmaniasis diagnostic test performance in Colombia and adapting the Western blot test in naturally and experimentally infected dogs. Methods Sera were obtained from 10 experimentally L. Infantum-infected dogs, 5 naturally infected dogs, 16 healthy dogs, 26 Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi and Leishmania (Viannia) spp infected dogs, 40 dogs from non-endemic areas and 150 from endemic areas. Sera were tested for L. infantum infection using immunofluorescent antibody (IFAT), ELISA and Western blot (WB) tests. Results Positives results were obtained for 73 % of known infected dogs by the IFAT test and false positives were obtained for 2.5 % of non-infected dogs using WB. ELISA was not efficient for diagnosis. 24 antigenic fractions were recognised in tested sera using WB; however, 29, 34, 50, 69, 75, 86, 99 and 123 kDa bands were recognised in sera from dogs from non-endemic areas, healthy dogs and Trypanosoma cruzi, Erhlichia canis, Dirofilaria immitis and Babesia canis infected dogs. The 13 kDa fraction proved potentially useful for diagnosing canine visceral leishmaniasis. ConclusionsThe separate use of parasitological and serological test could lead to misdiagnosis of Leishmania infection; using both kinds of technique simultaneously is thus highly recommended.

Objetivo Evaluar el desempeño de las pruebas empleadas en Colombia para el diagnóstico de la leishmaniasis visceral canina y adaptar una técnica de Western blot empleando animales experimental y naturalmente infectados. Metodología Se obtuvieron sueros de 10 perros infectados experimentalmente con L. infantum, 5 perros infectados naturalmente, 16 perros sanos, 26 de reacción cruzada (infectados con Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi, Leishmania (Viannia) spp.), 40 de zonas no endémicas y 150 de zona endémica. Todos fueron evaluados mediante las pruebas de inmunofluorescencia indirecta (IFI), ELISA y Western blot (WB). Resultados Se encontró que IFI tuvo el mayor porcentaje de positividad en los perros infectados (73 %) mientras que el menor porcentaje de falsos positivos se obtuvo por WB (2,5 %). La prueba de ELISA fue la menos eficiente. Fueron reconocidas 24 fracciones antigénicas, las bandas de 29, 34, 50, 69, 75, 86, 99 y 123 kDa fueron responsables de reacciones inespecíficas en los sueros de perros sanos, de zona no endémica y de reacción cruzada. Las bandas por debajo de 29 kDa mostraron ser potencialmente diagnósticas, especialmente la fracción de 13 kDa. Conclusiones Los métodos directos y serológicos pueden subdiagnosticar la infección por Leishmania, solamente un constructo que combine tanto pruebas directas como indirectas sería la forma más eficiente de diagnóstico.

Introducción. Los cultivos celulares de insectos son una metodología útil en estudios biomédicos y tecnológicos. Objetivo. El propósito principal del presente trabajo fue obtener y caracterizar cultivos celulares derivados de tejidos embrionarios de Aedes aegypti. Materiales y métodos. Se emplearon huevos embrionados para los explantes de tejidos en los medios de cultivos MM/VP12 y L-15/Grace, con suplemento de 20% de suero fetal bovino y una mezcla al 1% de antibiótico y antimicótico, con un rango de pH entre 6,8 y 7,0. Los cultivos se incubaron a una temperatura de 28ºC sin atmósfera de CO2. Resultados. El crecimiento celular se obtuvo en el medio L-15/Grace, 3 semanas después de haber sido sembrados los tejidos embrionarios; sin embargo, se necesitaron 6 meses para la formación de la monocapa confluente. Desde agosto de 2003 hasta junio de 2004, se habían realizado 28 subcultivos. Las células se caracterizaron morfológicamente; predominaron las formas epitelioides en subcultivos de pases altos. También se reconocieron las particularidades morfométricas del cariotipo y, además, se determinaron los perfiles isoenzimáticos y moleculares de los cultivos celulares, los cuales se compararon con muestras de adultos de la especie tomadas de la misma colonia y con líneas celulares derivadas de otros insectos. Discusión. Estas células representan, potencialmente, un importante sistema in vitro en investigaciones básicas y aplicadas.

Introduction. Cell cultures from insects are a useful methodology in technological and biomedical studies. Objective. The present work was aimed at obtaining and characterizing cell cultures derived from Aedes aegypti embryonic tissues. Materials and methods. Embryonated eggs were used for embryonic tissue explants in L-15/Grace and MM/VP12 culture media, supplemented with 20% fetal bovine serum and a mixture of 1% antimycotic and antibiotics, at a pH ranging from 6.8 to 7.0. The incubation temperature was 28º C; a CO2 atmosphere was not required. Results. Cell growth was obtained in L-15/Grace medium three weeks after embryonic tissues explants. Six months were required for achieving a confluent monolayer. Twenty eight serial cell subcultures were carried out from August 2003 to June 2004. Cell morphology was characterized as being epithelial in subcultures of high passages; karyotype morphometric characteristics were recognized and the molecular and isozymatic profiles were also established. The cultures were compared with adult samples from the species taken from the same colony and with cell lines derived from other insects. Discussion. These cells are an important in vitro system in applied and basic research.

Lutzomyia spinicrassa is a vector of Leishmania braziliensis in Colombia. This sand fly has a broad geographical distribution in Colombia and Venezuela and it is found mainly in coffee plantations. Baseline biological growth data of L. spinicrassa were obtained under experimental laboratory conditions. The development time from egg to adult ranged from 59 to 121 days, with 12.74 weeks in average. Based on cohorts of 100 females, horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (8.4 females per cohort female), generation time (12.74 weeks), intrinsic rate of population increase (0.17), and finite rate of population increment (1.18). The reproductive value for each class age of the cohort females was calculated. Vertical life tables were elaborated and mortality was described for the generation obtained of the field cohort. In addition, for two successive generations, additive variance and heritability for fecundity were estimated.

Con el propósito de establecer la prevalencia de leishmaniosis canina se efectuó un estudio descriptivo en 307 caninos, ubicados en 17 veredas de los municipios de Neiva, Tello y Algeciras, a los cuales se les practicó examen clínico, punción y aspiración del ganglio linfático poplíteo para extendido y coloración en lámina; toma de sangre completa por venopunción para análisis mediante la técnica de inmunofluorescencia indirecta. La población canina estuvo conformada por perros mestizos con edad promedio de tres años; el 30,6 % fueron hembras y el 69,4 % machos. En el examen físico de los caninos se observó: enflaquecimiento, 31 %; onicogrifosis, 29,3 %; linfadenitis de ganglio poplíteo 3,2 %; áreas alopécicas 21,8 %; lesiones eritematosas 11,5 % y úlceras cutáneas 3,3 %. El 1,4 % de los caninos analizados presentaron amastigotes en el aspirado y el 17,2 % fueron seropositivos. La prevalencia de anticuerpos contra L. chagasi es alta comparada con otros estudios realizados en Colombia y otros países, corroborando la endemicidad de la enfermedad en el área. Por consiguiente, es necesario fortalecer las estrategias de vigilancia epidemiológica y la búsqueda activa de casos humanos.

With the purpose of establishing the prevalence of canine leishmaniasis a descriptive study was carried out in 17 villages of the municipalities of Neiva, Tello and Algeciras, in the department of Huila, Colombia. The selected canine population was composed of 307 dogs, which were submitted to clinical examination, stained smears of lymph node needle aspiration, and bleeding for serological analysis by the Immunofluorescent Antibody Tests (IFAT). The canine population was conformed by mestizo dogs, with a 3 year age average; 30,6 % were females. Upon physical examination loss of weight was observed in 31% of dogs; lynfadenopathy of the popliteal node in 3,2 %; alopecia in 21,8 %, eritematous lesions in 11,5 % and skin ulcers in 3,3 %. 17,2 % of the dogs has a positive serology and in 1.43% of them, amastigotes were observed in the smears of lymph node aspirates. These prevalence of anti-L. chagasi antibodies is relatively high compared with other studies in Colombia and in other countries. The present study, confirms the endemic nature of the illness in the area, high lighting the need for strengthening public health surveillance and active search for human cases.

A new cell line, PC-0199-BR, was established from embryonated eggs of the mosquito Psorophora confinnis. To date (September 2000) it has had 62 continuous passages. This is the first report of a cell line of mosquitoes belonging to the genus Psorophora. Cell growth initially was achieved in the MM/VP12 medium, supplemented with 20% fetal bovine serum; however, the subcultures were later adapted to Grace's medium with 10% fetal bovine serum. Cell morphology in the primary cultures was heterogeneous; but later in the established cell line, the predominant cell type was epithelioid. Cultured cells were predominantly diploid (2n=6); however, chromosome abnormalities were observed in a small proportion of the cells in later passages. C and G band patterns were also determined in the karyotype. The cell line isozyme profiles coincided with pupae and adult samples of the species taken from the same colony. A preliminary arbovirus susceptibility study for the cell line was undertaken. No evidence was observed of contamination of the cell line with bacteria, fungi or mycoplasma.