Phlebotomine sandflies (Diptera: Psychodidae) collected in rural area from San Bernardo del Viento (Córdoba, Colombia) Flebotomíneos (Diptera: Psychodidae) colectados en área rural de San Bernardo del Viento (Córdoba Colombia)

Objective. Our aim was evaluate the presence of phlebotomine sandflies in an important rural area with eco-epidemiological interest in San Bernardo del Viento (Córdoba, Colombia). Materials and Methods. The insects identified were collected using CDC-traps and manual aspirator in closed trees between May to August 2015. The specimens were conserved and fragmented for clarification of internal structures and identification of morphological keys. Results. Three species of phlebotomine sand flies were identified: Lutzomyia gomezi (Nitzulescu 1931), Pintomyia rangeliana (Ortiz 1952) y Micropygomyia cayennensis (Floch & Abonnenc 1941). Conclusions. The presence of L. gomezi, main vector of cutaneous leishmaniasis in Colombia, is a risk factor for rural people from the zone.


INTRODUCTION
Sandflies are small blood-feeding dipterous belonging mainly to Phlebotominae subfamily (Diptera: Psychodidae) -genus Lutzomyia in the new world, and containing approximately >400 species, of which 153 have been found in Colombia (1,2). These hematophagous insects are primary vectors of protozoa from genus Leishmania (Ross, 1903), causative agents of leishmaniasis, an infectious disease with high prevalence in Colombia, registering >17.000 annual cases (3). The visceral and cutaneous leishmaniasis cases reported at the Cordoba Department were 229 cases until 52th week of 2015 (4). This high prevalence of leishmaniasis is due to convergence reservoirs, vectors and susceptible humans host among a geographic zone that cross the departments of Cordoba, Bolivar and Sucre called "Montes de María" (5). The fauna of sand flies in Cordoba is poorly known and 13 species have been recorded in last studies (2,7). However, these registers were performed in endemic focus of leishmaniasis in municipalities as San Andres de Sotavento, Cereté, Chinú and Sahagún (1,6,7); highlighting the presence of the vectors of visceral leishmaniasis, Lutzomyia longipalpis (Lutz & Neiva 1912) and Pintomyia evansi (Nuñez-Tovar 1924).
However, further studies should be performed because they are necessary to establish the presence of phlebotomine sand flies species in other regions of Cordoba department. Even though these regions do not appears to be epidemiologically important for active disease.
Our aim was evaluate the presence of phlebotomine sandflies in an important rural area with eco-epidemiological interest in San Bernardo del Viento (Córdoba, Colombia), where there are not records of these insects and human cases of leishmaniasis. The species identified are discussed in the context of leishmaniasis and potential risk of transmission in the studied zone.

MATERIALS AND METHODS
Site estudy. San Bernardo del Viento (Córdoba, Colombia), is a municipality located in the Colombian Caribbean coast (approximate population 32.000) and is two meters above sea level, 30°C mean temperature, and is bordered by delta of Sinú River. The locality of our study is called "Chiquilito" (9°19´59.88" N, 75°58´0.12" W) and consist of rice fields, pastures and mangos; there is a high diversity wildlife, include 296 species of birds (belonging to 61 families, at which 64 species are migratory); reptiles and
Nuestro objetivo era evaluar la presencia de flebotomíneos en un área rural de interés ecoepidemiológico perteneciente al municipio de San Bernardo del Viento (Córdoba, Colombia), donde no hay registros de estos insectos ni de caos de leishmaniasis humana. Las especies identificadas se discuten en el contexto de la leishmaniasis y el riesgo potencial de transmisión en la zona de estudio.

Samples collection.
Captures were performed in the municipality of San Bernardo del Viento, from May to August 2015. Sandflies were collected in forested areas impacted by the presence of man around residences, and around the rice crops present in the peri-domicile, using CDC light traps during 14-hour (5:00 p.m. to 7:00 a.m.). Manual and manual aspirators were only used in resting sites of sandflies from 6:00 a.m. to 9:00 a.m.
Preparation. Preparation of the sandflies was performed individually using lacto-phenol and were identified to species level using microscopic observation of various internal/ external morphological characteristics, using the keys proposed by Young and Duncan (9).

RESULTS
A total of 78 phlebotomine sandflies belonging to three species and Lutzomyia genus were

DISCUSSION
M. cayennensis was predominant species captured in the area the study. This species is characterized by feeding on rodents and reptiles (9) and its high capacity for exploitation new resources in farm zones. Also, it is collected in areas closed to human houses, domestic animals and forest relicts (5,10-12). For instance, females of this species were collected in Ovejas (Department of Sucre, Colombia) and have been found infected naturally with Trypanosomatidae parasites using polymerase chain reaction for ssRNA (small subunit of RNA ribosomal) (5). Despite its zoophilic behavior, recent authors highlight that those species have high population densities in peaks of activity-bite in the range of 6:00 p.m. -10:00 p.m., evidencing endophilic preferences and being associated with rural and urban environments in localities of Córdoba, Sucre and Bolivar (10-12).
P. rangeliana was a second species captured in order of abundance; this species has been considered moderately anthropophilic and founded infected with flagellate promastigotes belonging to Leishmania sp. identified like as Leishmania venezuelensis (Bonfante-Garrido 1980) (13), the etiologic agent of cutaneous leishmaniasis in Venezuela.
In third place, L. gomezi is an anthropophilic species with endophilic behavior behavior on human settlements and have been considered probably vector of cutaneous leishmaniasis in several regions of Colombia (2,5,7,(10)(11)(12). This phlebotomine sand-fly has been detected naturally infected with Leishmania panamensis (Lainson & Shawn 1972) in Boyacá (14) and Leishmania braziliensis (Viannia 1911) in Venezuela. Also, has been found infected with promastigotes not identified in Panama, Ecuador and Colombia (10). L. gomezi is a vector with high adaptive abilities (ecological plasticity) being associated with disturbing environments and low abundances in forest areas, suggesting as species adapted to deforested zones, where is find a diversity of food resources (10)(11)(12)15). The records of species belonging to phlebotomine sandflies in San Bernardo del Viento (Córdoba, Colombia), have an epidemiological relevance
En tercer lugar, L. gomezi es una especie antropofílica y de comportamiento endofílico en asentamientos humanos, y ha sido considerada como un probable vector de leishmaniasis cutánea en varias regiones de Colombia (2,5,7,(10)(11)(12). Este flebotomíneo ha sido detectado con infección natural por Leishmania panamensis (Lainson & Shawn 1972) en Boyacá (14) y Leishmania braziliensis (Viannia 1911) en Venezuela. También se ha encontrado infectad con promastigotes no identificados en Panamá, from the point of view of leishmaniasis, also provides important information about the geographic distribution and extensive modeling about ecological niche and potential risk of transmission, allowing determine possible active zones and indicate ecological key-factors for presence of leishmaniasis. In this sense, we might to perform a taxonomic inventory more extensive in that municipality because is very probably that biodiversity increases, also is fundamental understand patterns of population fluctuations in species of genus Lutzomyia/Pintomyia, and natural infection rates with Leishmania parasites.

Acknowledgments
This work was supported by Departamento Administrativo de Ciencia, Tecnología e Innovación -Colciencias with grant project 111549326198. Richard Hoyos López was recipient of a doctoral fellowship from the "Programa de Doctorados Nacionales -Colciencias" (Convocatoria 528).