Diseño y evaluación de una trampa dirigida a la vigilancia entomológica de hembras de aedes aegypti (díptera : culicidae) utilizando cebos de origen microbiano.

Tesis (Doctorado en Ciencias con Acentuación en Entomología Médica) UANL, 2012.

Hongos entomopatógenos (mycota : deuteromycetes) aislados en el Noreste de México : impacto sobre la longevidad, fecundidad, fertilidad y tasas de cópula e inseminación en aedes aegypti (diptera-culicidae).

Tesis (Doctor en Ciencias con Especialidad en Entomología Médica) UANL, 2011.

Caracterización de cepas de bacillus thuringiensis y su toxicidad contra tres poblaciones de aedes aegypti.

Tesis (Doctor en Ciencias con Especialidad en Biotecnología) UANL, 2010.

Perfil de resistencia y mutación kdr asociada a insecticidas piretroides en aedes aegypti (L.) (DIPTERA: CULICIDAE) de Veracruz, México.

Tesis (Doctor en Ciencias con Acentuación en Entomología Médica) UANL, 2010.

Caracterización bioquímica y molecular de subpoblaciones silvestres de Aedes aegypti (L) de Venezuela sometidas a presión con insecticidas de uso común.

Tesis (Doctor en Ciencias con Acentuación en Entomología Médica) UANL, 2012.

Potencial de transmisión de dengue en zonas no domiciliares a través de la búsqueda de hembras Aedes aegypti infectadas por virus del dengue en el área metropolitana de Monterrey, Nuevo León, México.

Tesis (Doctor en Ciencias con Acentuación en Entomología Médica) UANL, 2012

Infección potencial secundaria a dengue asociada a la presencia de poblaciones de Aedes aegypti (Linnaeus, 1762) en áreas domiciliares de casos confirmados de dengue clásico y dengue hemorrágico en Monterrey, México.

Tesis (Doctor en Ciencias con Acentuación en Entomología Médica) UANL, 2012

Resistencia a insecticidas en el mosquito vector del dengue Aedes aegypti (L) en dos épocas de transmisión de la enfermedad en Mérida, Yucatán.

Tesis (Doctorado en Ciencias con Acentuación en Entomología Médica) UANL, 2013.

Análisis de la susceptibilidad de una línea celular de Aedes aegypti (Diptera: Culicidae) a la infección con Leishmania (L) chagasi y Leishmania (V) braziliensis

Se evaluó la susceptibilidad de los cultivos celulares derivados de tejidos embrionarios de Aedes aegypti a la infección con Leishmania (L) chagasi y Leishmania (V) braziliensis, agentes etiológicos de leishmaniasis visceral americana y leishmaniasis cutánea, respectivamente. Metodología: Se seleccionaron células de A. aegypti mantenidas en una mezcla de medio de cultivo Grace/L15, suplementado con suero fetal bovino al 15%, albendazol 5,4 mg/ml y una mezcla de antibióticos, e incubadas a una temperatura promedio de 26 °C. Los cultivos celulares fueron inoculados con promastigotes metacíclicos de la cepa MH/CO/84/CI-044B de L. chagasi y la cepa HOM/BR752903 de L. braziliensis en una concentración de 10 parásitos por célula. Como control positivo de la infección se utilizó la línea celular J774. Resultados: Los registros más altos en el porcentaje de infección y en el número de amastigotes por células en los cultivos celulares A. aegypti y en la línea celular J774 se obtuvieron en los días 6 y 9 pos-infección. Los resultados mostraron interacción, internalización y maduración in vitro de las dos especies del parásito en las células de este insecto no vector de Leishmania. Las células de A. aegypti infectadas mostraron cambios en el área por la influencia de los parásitos, contrario a lo registrado en las células no infectadas (P<0,05). Conclusión: Los cultivos celulares de A. aegypti emergen como un nuevo modelo in vitro para el estudio del ciclo biológico de L. chagasi y L. braziliensis.

Evaluation of the Toxic Effect from Eupatorium Microphyllum L.F. (Asteraceae) Extracts on Aedes Aegypti Larvae (Diptera: Culicidae) in Laboratory Conditions

In the present work the toxic activity of extracts of Eupatorium microphyllum L.F. was evaluated on 4th 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-1and 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-1with 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.

Caracterização funcional do gene da Glutationa-S-Transferase Epsilon 2 (GSTE2) em Aedes aegypti

As Glutationa-S-Transferases (GSTs) em insetos desempenham um papel fundamental na metabolização de inseticidas químicos, e provavelmente estão envolvidas na proteção contra o estresse oxidativo decorrente da exposição a xenobióticos. O objetivo do trabalho foi a caracterização funcional do gene GSTE2 em linhagens de Aedes aegypti com diferentes perfis de susceptibilidade ao temephos. Foram usadas uma colônia susceptível (RecLab) e outra resistente, (RecR). Larvas de ambas as linhagens foram divididas em dois grupos: exposto ao temephos com concentrações subletais e não exposto. Os indivíduos sobreviventes foram usados em ensaios enzimáticos para medir a atividade das GSTs totais contra os substratos CDNB (padrão) e o 4-HNE, um produto endógeno resultante da peroxidação de lipídeos. Adicionalmente, foi feito o sequenciamento do cDNA deste gene em amostras das duas linhagens e a sua expressão foi investigada. A GSTE2 das duas linhagens foi expressa em sistema heterólogo e purificada para avaliação da atividade metabólica contra o 4-HNE, através de testes de biocatálise. Os resultados revelaram que a atividade enzimática da GST usando o CDNB foi normal para RecLab, em ambas as condições estudadas, porém, para RecR houve alteração na atividade de GST, para os dois grugo estudados . Usando o 4-HNE como substrato, as duas linhagens apresentaram um perfil enzimático alterado para GST em relação à Rock, com uma resposta aumentada após a exposição ao temephos. Foram identificados polimorfismos que diferenciam as duas linhagens. Os resultados de expressão gênica indicaram que as larvas resistentes apresentam níveis de expressão significativamente maiores do que as susceptíveis, e em RecR a expressão caiu após a exposição, sugerindo o envolvimento dessa enzima nos processos de resistência metabólica na linhagem RecR. Esses dados abrem novas perspectivas de monitoramento da resistência metabólica em Ae. aegypti

Effects of Plasmodium gallinaceum on hemolymph physiology of Aedes aegypti during parasite development

Insect disease vectors show diminished fecundity when infected with Plasmodium. This phenomenon has already been demonstrated in laboratory models such as Aedes aegypti, Anopheles gambiae and Anopheles stephensi. This study demonstrates several changes in physiological processes of A. aegypti occurring upon infection with Plasmodium gallinaceum, such as reduced ecdysteroid levels in hemolymph as well as altered expression patterns for genes involved in vitellogenesis, lipid transport and immune response. Furthermore, we could show that P. gallinaceum infected A. aegypti presented a reduction in reproductive fitness, accompanied by an activated innate immune response and increase in lipophorin expression, with the latter possibly representing a nutritional resource for Plasmodium sporozoites. (C) 2010 Elsevier Ltd. All rights reserved.

A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombin coagulant inhibitory activity

Kazal-type inhibitors play several important roles in invertebrates, such as anticoagulant, vasodilator and antimicrobial activities. Putative Kazal-type inhibitors were described in several insect transcriptomes. In this paper we characterized for the first time a Kazal unique domain trypsin inhibitor from the Aedes aegypti mosquito. Previously, analyses of sialotranscriptome of A. aegypti showed the potential presence of a Kazal-type serine protease inhibitor, in female salivary glands, carcass and also in whole male, which we named AaTI (A. aegypti trypsin inhibitor). AaTI sequence showed amino acid sequence similarity with insect thrombin inhibitors, serine protease inhibitor from Litopenaeus vannamei hemocytes and tryptase inhibitor from leech Hirudo medicinalis (LDTI). In this work we expressed, purified and characterized the recombinant AaTI (rAaTI). Molecular weight of purified rAaTI was 7 kDa rAaTI presented dissociation constant (K(i)) of 0.15 and 3.8 nM toward trypsin and plasmin, respectively, and it weakly inhibited thrombin amidolytic activity. The rAaTI was also able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. AaTI transcription was confirmed in A. aegypti female salivary gland and gut 3 h and 24 h after blood feeding, suggesting that this molecule can act as anticoagulant during the feeding and digestive processes. Its transcription in larvae and pupae suggested that AaTI may also play other functions during the mosquito`s development. (C) 2010 Elsevier Masson SAS. All rights reserved.

The Aedes aegypti larval transcriptome: a comparative perspective with emphasis on trypsins and the domain structure of peritrophins

The genome sequence of Aedes aegypti was recently reported. A significant amount of Expressed Sequence Tags (ESTs) were sequenced to aid in the gene prediction process. In the present work we describe an integrated analysis of the genomic and EST data, focusing on genes with preferential expression in larvae (LG), adults (AG) and in both stages (SG). A total of 913 genes (5.4% of the transcript complement) are LG, including ion transporters and cuticle proteins that are important for ion homeostasis and defense. From a starting set of 245 genes encoding the trypsin domain, we identified 66 putative LG, AG, and SG trypsins by manual curation. Phylogenetic analyses showed that AG trypsins are divergent from their larval counterparts (LG), grouping with blood-induced trypsins from Anopheles gambiae and Simulium vittatum. These results support the hypothesis that blood-feeding arose only once, in the ancestral Culicomorpha. Peritrophins are proteins that interlock chitin fibrils to form the peritrophic membrane (PM) that compartmentalizes the food in the midgut. These proteins are recognized by having chitin-binding domains with 6 conserved Cys and may also present mucin-like domains (regions expected to be highly O-glycosylated). PM may be formed by a ring of cells (type 2, seen in Ae. aegypti larvae and Drosophila melanogaster) or by most midgut cells (type 1, found in Ae. aegypti adult and Tribolium castaneum). LG and D. melanogaster peritrophins have more complex domain structures than AG and T. castaneum peritrophins. Furthermore, mucin-like domains of peritrophins from T. castaneum (feeding on rough food) are lengthier than those of adult Ae. aegypti (blood-feeding). This suggests, for the first time, that type 1 and type 2 PM may have variable molecular architectures determined by different peritrophins and/or ancillary proteins, which may be partly modulated by diet.