Prevalence of canine visceral leishmaniasis in municipalities of huila, colombia

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.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

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Insect cell cultures are an important biotechnological tool for basic and applied studies. The objective of this work was to establish and characterise a new cell line from Culex quinquefasciatus embryonic tissues. Embryonated eggs were taken as a source of tissue to make explants that were seeded in L-15, Grace's, Grace's/L-15, MM/VP12, Schneider's and DMEM culture media with a pH range from 6.7-6.9 and incubated at 28ºC. The morphological, cytogenetic, biochemical and molecular characteristics of the cell cultures were examined by observing the cell shapes, obtaining the karyotypes, using a cellulose-acetate electrophoretic system and performing random amplified polymorphic DNA-polymerase chain reaction analysis, respectively. The Grace's/L-15 medium provided the optimal nutritional conditions for cell adhesion and proliferation. Approximately 40-60 days following the explant procedure, a confluent monolayer was formed. Cellular morphology in the primary cultures and the subcultures was heterogeneous, but in the monolayer the epithelioid morphology type predominated. A karyotype with a diploid number of six chromosomes (2n = 6) was observed. Isoenzymatic and molecular patterns of the mosquito cell cultures matched those obtained from the immature and adult forms of the same species. Eighteen subcultures were generated. These cell cultures potentially constitute a useful tool for use in biomedical applications

Experimental infection of Leishmania (L.) chagasi in a cell line derived from Lutzomyia longipalpis (Diptera:Psychodidae)

The present work describes the in vitro infection of a cell line Lulo, derived from Lutzomyia longipalpis embryonic tissue, by Leishmania chagasi promastigotes. This infection process is compared with a parallel one developed using the J774 cell line. The L. chagasi MH/CO/84/CI-044B strain was used for experimental infection in two cell lines. The cells were seeded on glass coverslips in 24-well plates to reach a final number of 2 x 10(5) cells/well. Parasites were added to the adhered Lulo and J774 cells in a 10:1 ratio and were incubated at 28 and 37ºC respectively. After 2, 4, 6, 8, and 10 days post-infection, the cells were extensively washed with PBS, fixed with methanol, and stained with Giemsa. The number of internalized parasites was determined by counting at least 400 cultured cells on each coverslip. The results showed continuous interaction between L. chagasi promastigotes with the cell lines. Some ultrastructural characteristics of the amastigote forms were observed using transmission electron microscopy. The highest percentage of infection in Lulo cells was registered on day 6 post-infection (29.6%) and on day 4 in the J774 cells (51%). This work shows similarities and differences in the L. chagasi experimental infection process in the two cell lines. However, Lulo cells emerge as a new model to study the life-cycle of this parasite