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Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae): Nuevos cultivos celulares de Aedes aegypti
Ariosto,Ardila; Escovar,Jesús; Bello,Felio;
Biomédica , 2005,
Abstract: 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.
Establecimiento, mantenimiento y productividad de una colonia de laboratorio de Lutzomyia spinicrassa Morales, Osorno-Mesa, Osorno y Hoyos, 1969 (Diptera: Psychodidae) en Colombia
Morales,Alberto; Bello,Felio; Cárdenas,Estrella;
Revista Ciencias de la Salud , 2005,
Abstract: lutzomyia spinicrassa is a vector of leishmania braziliensis. this sand fly has a broad geographical distribution in colombia and venezuela and it's founded mainly in coffee plantations. methodology: starting from 600 females of l. spinicrassa captured in field a laboratory colony was established. the development time from egg to adult ranged from 58 to 78 days, 11 weeks in average. population parameters of five successive generations maintained in groups were compared with a generation reared individually. results: the following parameters were obtained in each experimental condition: net rate of reproduction (6.92 and 7 females per female per generation), intrinsic rate of population increment (0.17 and 0.18 females per female per week) and finite rate of population increment (1.06 and 1.19 individuals per female per week). conclusion: these data suggest that the colony of l. spinicrassa had a constant increment during the six analyzed generations.
Evaluación del efecto tóxico de extractos de Eupatorium microphyllum L.F. (Asteraceae) sobre larvas de Aedes aegypti (Diptera: Culicidae) en condiciones de laboratorio
Rozo,álvaro; Zapata,Cristina; Bello,Felio J.;
Revista Ciencias de la Salud , 2008,
Abstract: in the present work the toxic activity of extracts of eupatorium microphyllum l.f. was evaluated on 4 th instar larvae of the mosquito aedes aegypti (linneaus), under laboratory conditions. aqueous extracts were utilized in concentrations of 500 mg l-1, 1,500 mg l-1 and 2,500 mg l-1 and acetone in concentrations of 10 mg l-1, 20 mg l-1, 30 mg l-1, 40 mg l-1 and 50 mg l-1. the bioassays were carried out for triplicate each one with 20 larvae, exposed for 24 hours to 150 ml of solution. in all the bioassays were employed control groups. in the evaluation of the acetone extracts, a negative control was employed to avoid that the mortality of the larvae to occur on account of the solvent. the aqueous extracts showed low moderate action in the mortality of larvae, less than 20%. on the contrary, the action of the acetone extracts was observed to 10 and 20 mg l-1 with 15% of mortality, while to 30 and 40 mg l-1 were registered 22 to 38% of mortality. however, to 50 mg l-1 the mortality was of 95.4% with highly significant statistical results. the concentrations of the acetone extracts showed to be the most efficient for the control of the mosquitoes selected. both types of extracts showed toxic effect in larvae of a. aegypti, nevertheless, greater effect in the acetone extracts was observed relating to the aqueous extracts of e. microphyllum, which constitutes a viable alternative in the search of new larvicides from composed natural.
Evaluation of the Toxic Effect from Eupatorium Microphyllum L.F. (Asteraceae) Extracts on Aedes Aegypti Larvae (Diptera: Culicidae) in Laboratory Conditions
álvaro Rozo,Cristina Zapata,Felio J. Bello
Revista Ciencias de la Salud , 2008,
Abstract: In the present work the toxic activity ofextracts of Eupatorium microphyllum L.F. wasevaluated on 4th instar larvae of the mosquitoAedes aegypti (Linneaus), under laboratoryconditions. Aqueous extracts were utilized inconcentrations of 500 mg L-1, 1,500 mg L-1 and2,500 mg L-1 and acetone in concentrations of10 mg L-1, 20 mg L-1, 30 mg L-1, 40 mg L-1and50 mg L-1. The bioassays were carried out fortriplicate each one with 20 larvae, exposedfor 24 hours to 150 mL of solution. In all thebioassays were employed control groups. Inthe evaluation of the acetone extracts, a negativecontrol was employed to avoid that themortality of the larvae to occur on account ofthe solvent. The Aqueous extracts showed lowmoderate action in the mortality of larvae, lessthan 20%. On the contrary, the action of theacetone extracts was observed to 10 and 20 mgL-1with 15% of mortality, while to 30 and 40mg L-1 were registered 22 to 38% of mortality.However, to 50 mg L-1 the mortality wasof 95.4% with highly significant statisticalresults. The concentrations of the acetone extractsshowed to be the most efficient for thecontrol of the mosquitoes selected. Both typesof extracts showed toxic effect in larvae of A.aegypti, nevertheless, greater effect in theacetone extracts was observed relating to theaqueous extracts of E. microphyllum, whichconstitutes a viable alternative in the search ofnew larvicides from composed natural.
CARACTERíSTICAS DE CULTIVOS CELULARES PRIMARIOS DERIVADOS DE Sarconesiopsis magellanica (LE GUILLOU, 1842) (DIPTERA: CALLIPHORIDAE) PRIMARY CELL CULTURES DERIVED FROM Sarconesiopsis magellanica (LE GUILLOU, 1842) (DIPTERA: CALLIPHORIDAE)
Mónica Cruz B.,Felio J. Bello
Revista U.D.C.A Actualidad & Divulgación Científica , 2012,
Abstract: El objetivo principal del presente trabajo fue obtener cultivos celulares primarios derivados de tejido embrionario de Sarconesiopsis magellanica (Diptera: Calliphoridae), mosca importante por sus aplicaciones en el establecimiento del intervalo post-mortem. Se evaluaron siete medios de cultivo (Grace, Grace/L15, MM, VP12, MM/VP12, Eagle y Schneider), suplementados con 20% de suero fetal bovino. Se observó adhesión, crecimiento y proliferación celular en los medios L15, Schneider y Grace/L15, lográndose mejores resultados en los dos últimos. La obtención de la monocapa confluente, se presentó en un tiempo promedio de doce días, después de realizados los explantes. El patrón de crecimiento de los cultivos primarios mostró la presencia de vesículas con células adheridas a sus paredes, las cuales, ayudaron a la formación de la monocapa confluente. La morfología celular predominante en la monocapa correspondió a formas fibroblastoides y, en menor proporción, a epitelioides, mostrando las primeras una apariencia similar a células nerviosas. Adicionalmente, se registraron en los primeros días de los cultivos primarios, especialmente, en algunos conglomerados de células, movimientos contráctiles semejantes a los realizados por células musculares. Estos cultivos celulares primarios derivados de S. mangellanica representan, potencialmente, sustratos adecuados para realizar ensayos posteriores de susceptibilidad a infección con arbovirus y parásitos. The present work was aimed at obtaining and characterizing cell cultures from Sarconesiopsis magellanica embryonic tissue (Diptera: Calliphoridae). This fly is important by its applications in the establishment of post-mortem interval. This study evaluated seven culture medium (Grace, Grace/L15, MM, VP12, MM/VP12, Eagle and Schneider), supplemented with 20% fetal bovine serum. The adhesion, cell growth and proliferation were observed in L15 medium, Schneider and Grace/L15, achieving better results in the last two. The confluent monolayer was present in an average of twelve days after that the explants were made. The growth pattern of the primary cultures showed the presence of vesicles with cells adhering to its walls, which helped to form a confluent monolayer. The predominant cell morphology in monolayer correspond to fibroblastoid a lesser extent to epithelioid, showing the first similar to nerve cells. Additionally, there was in the early days of primary cultures, especially in some clusters of cells, contractile movement, similar to those made by muscle cells. These primary cell cultures derived from S. magellan
Establishment, maintenance and productivity of a colony of laboratory from Lutzomyia spinicrassa Morales, Osorno-Mesa, Osorno & Hoyos, 1969 (Diptera: Psychodidae) in Colombia.
Alberto Morales*, Felio Bello?, Estrella Cárdenas,Felio Bello,Estrella Cárdenas
Revista Ciencias de la Salud , 2005,
Abstract: Lutzomyia spinicrassa is a vector of Leishmaniabraziliensis. This sand fly has a broadgeographical distribution in Colombia and Venezuelaand it’s founded mainly in coffeeplantations. Methodology: Starting from 600females of L. spinicrassa captured in field alaboratory colony was established. The developmenttime from egg to adult ranged from 58 to78 days, 11 weeks in average. Population parametersof five successive generations maintainedin groups were compared with a generationreared individually. Results: The followingparameters were obtained in each experimentalcondition: net rate of reproduction (6.92 and 7females per female per generation), intrinsic rate of population increment (0.17 and 0.18 females perfemale per week) and finite rate of populationincrement (1.06 and 1.19 individuals per femaleper week). Conclusion: These data suggest thatthe colony of L. spinicrassa had a constant incrementduring the six analyzed generations.
Establishment and characterization of a new continuous cell line from Lutzomyia longipalpis (Diptera: Psychodidae) and its susceptibility to infections with arboviruses and Leishmania chagasi
Rey, Gloria J;Ferro, Cristina;Bello, Felio J;
Memórias do Instituto Oswaldo Cruz , 2000, DOI: 10.1590/S0074-02762000000100017
Abstract: embryonic tissue explants of the sand fly lutzomyia longipalpis (lutz & neiva 1912) the main vector of leishmania chagasi (cunha and chagas), were used to obtain a continuous cell line (lulo). the tissues were seeded in mm/vp12 medium and these were incubated at 28oc. the first subculture was obtained 45 days after explanting and 96 passages have been made to date. lulo is composed of epithelioid cells, showed a 0.04 generations/hour exponential growth rate and population doubling time at 24.7 h. the cell line isoenzymatic profiles were determined by using pgi, pgm, mpi and 6-pgdh systems, coinciding with patterns obtained from the same species and colony's pupae and adults. the species karyotype characteristics were recognized (2n = 8), in which pair 1 is subtelocentric and pairs 2, 3 and 4 are metacentric. lulo was free from bacterial, fungal, mycoplasmic and viral infection. susceptibility to five arbovirus was determined, the same as lulo interaction with leishmania promastigotes.
Análisis de la susceptibilidad de una línea celular de Aedes aegypti (Diptera: Culicidae) a la infección con leishmania (L) chagasi y Leishmania (V) braziliensis
Mu?oz-Camargo,Carolina; Barreto,Alfonso; Bello,Felio;
Revista Ciencias de la Salud , 2005,
Abstract: the susceptibility of culture cells derived from embryonic tissues of aedes aegypti to the infection with leishmania (l) chagasi and leishmania (v) braziliensis was evaluated. methodology: these parasites are etiological agents of american visceral leishmaniasis and cutaneous leishmaniasis, respectively. selected cells of aedes aegypti were maintained in culture medium grace/l15, supplement with 15% bovine fetal serum, 5,4 mg/ml of albendazol and an antibiotic mixture and incubated at an average temperature of 26°c. the cultures were inoculated with metacyclic promastigotes of the strain mh/ co/84/ci-044b of l. chagasi and the strain hom/br752903 of l. braziliensis in a concentration of 10 parasites by cell. the j774 cell line was used as positive control of infection. results: the highest percentage of infection represented as the number of amastigotes per cell in a. aegyti cell cultures and in the j774 cell line were obtained on days 6 and 9 post-infection. the results showed interaction, internalization and maturation in vitro of the two species of the parasite in the cells of a non-vector insect of leishmania. infected a. aegypti cells showed changes in its area because of the influence of the parasites that differ significantly (p <0.05) compared to not infected cells. conclusion: cell cultures from a. aegypti emerge as a new in vitro model for the study of the biological cycle of l. chagasi and l. braziliensis.
Susceptibility Analysis of a Cell Line Derived from Aedes aegypti (Diptera: ulicidae)
Carolina Mu?oz-Camargo,Alfonso Barreto,Felio Bello
Revista Ciencias de la Salud , 2005,
Abstract: The susceptibility of culture cells derived fromembryonic tissues of Aedes aegypti to theinfection with Leishmania (L) chagasi andLeishmania (V) braziliensis was evaluated.Methodology: These parasites are etiologicalagents of American visceral leishmaniasis andcutaneous leishmaniasis, respectively. Selectedcells of Aedes aegypti were maintained in culturemedium Grace/L15, supplement with 15% bovinefetal serum, 5,4 mg/ml of albendazol and anantibiotic mixture and incubated at an average temperature of 26°C. The cultures were inoculatedwith metacyclic promastigotes of the strain MH/CO/84/CI-044B of L. chagasi and the strainHOM/BR752903 of L. braziliensis in a concentrationof 10 parasites by cell. The J774 cellline was used as positive control of infection.Results: The highest percentage of infectionrepresented as the number of amastigotes per cellin A. aegyti cell cultures and in the J774 cell linewere obtained on days 6 and 9 post-infection.The results showed interaction, internalizationand maturation in vitro of the two species of theparasite in the cells of a non-vector insect ofLeishmania. Infected A. aegypti cells showedchanges in its area because of the influence of theparasites that differ significantly (P <0.05)compared to not infected cells. Conclusion: Cellcultures from A. aegypti emerge as a new in vitromodel for the study of the biological cycle of L.chagasi and L. braziliensis.
Trypanosoma rangeli infected mouse sera reactivity with Trypanosoma cruzi synthetic peptides
CLAUDIO ZUNIGA,RAMON VARGAS,MARIA TERESA PALAU,FELIO BELLO
Parasitología latinoamericana , 2007,
Abstract: In man, differential diagnosis of Trypanosoma cruzi and Trypanosoma rangeli infections represents a serious problem, not only because both parasites present similar geographical distribution, the same hosts and sometimes the same insect vector, but also because they have common antigen determinants. In this work IgM and IgG humoral responses to T. cruzi syntethic peptides in mice infected with T. cruzi and with T. rangeli were analysed. In a immunoradiometric assay (IRMA ) 6 syntethic peptides were used, denominated as clones 1, 2, SAPA, 13, 30 and 36. The results showed that sera from infected mice with T. rangeli recognized all peptides derived from T. cruzi proteins, at IgM as well as IgG level. Reactivity with peptide SAPA is discussed as previous work indicated that SAPA is not codified in the T. rangeli genome. Our results support the suggestion that crossed reactions are due to the fact that both parasites present common antigens
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