RESUMEN
Malaria is a serious public health problem, the control of which involves actions directed against its vector, mosquitoes of the genus Anopheles Meigan, 1818. The study of insect biology, ecology, and behavior is simplified when these insects are maintained at the laboratory. However, many of the species are eurygamic and require induced mating. Females of Anopheles marajoara Galvão e Damasceno, 1942 were collected at Mazagão county, State of Amapá, Brazil. F1 eggs were obtained through forced oviposition and raised until mosquito emergence. Around 300 mosquitoes were maintained in each cage and were fed with a 10% sugar solution. Induced mating was made to obtain the other generations. Females had their spermathecae examined for the presence of sperm. The efficacy of coupling in each generation was evaluated. The viability of a sample of generations F5, F9, F12, and F14 was followed from larvae to adult. Two free mating attempts were done. The results demonstrate adaptation of An. marajoara to laboratory conditions over 21 generations, with viability rates temporally increasing. There was no evidence of adaptation to free mating.
Asunto(s)
Animales de Laboratorio/fisiología , Anopheles/fisiología , Insectos Vectores/fisiología , Conducta Sexual Animal , Animales , Brasil , Femenino , Malaria , MasculinoRESUMEN
The constitutive ribosomal gene rp49 is frequently used as an endogenous control in Drosophila gene expression experiments. Using the degenerate primer PCR technique we have cloned a fragment homologous to this gene in Anopheles aquasalis Curry, a Neotropical vector of malaria. In addition, based on this first sequence, a new primer was designed, which allowed the isolation of fragments of rp49 in two other species, Aedes aegypti (Linnaeus) and Culex quinquefasciatus Say, suggesting that it could be used to clone fragments of this gene in a number of other mosquito species. Primers were also designed to specifically amplify rp49 cDNA fragments in An. aquasalis and Ae. aegypti, showing that rp49 could be used as a good constitutive control in gene expression studies of these and other vectorially important mosquito species.
Asunto(s)
Culicidae/genética , Proteínas de Drosophila/aislamiento & purificación , Drosophila/genética , Insectos Vectores/genética , Proteínas Ribosómicas/aislamiento & purificación , Aedes/genética , Animales , Anopheles/genética , Culex/genética , Cartilla de ADN/genética , ADN Complementario/genética , Proteínas de Drosophila/genética , Expresión Génica , Reacción en Cadena de la Polimerasa , Proteínas Ribosómicas/genéticaRESUMEN
The constitutive ribosomal gene rp49 is frequently used as an endogenous control in Drosophila gene expression experiments. Using the degenerate primer PCR technique we have cloned a fragment homologous to this gene in Anopheles aquasalis Curry, a Neotropical vector of malaria. In addition, based on this first sequence, a new primer was designed, which allowed the isolation of fragments of rp49 in two other species, Aedes aegypti (Linnaeus) and Culex quinquefasciatus Say, suggesting that it could be used to clone fragments of this gene in a number of other mosquito species. Primers were also designed to specifically amplify rp49 cDNA fragments in An. aquasalis and Ae. aegypti, showing that rp49 could be used as a good constitutive control in gene expression studies of these and other vectorially important mosquito species.
Asunto(s)
Animales , Aedes/genética , Anopheles/genética , Culex/genética , Proteínas de Drosophila/aislamiento & purificación , Drosophila/genética , Insectos Vectores/genética , ADN Complementario/genética , Proteínas de Drosophila/genética , Expresión Génica , Reacción en Cadena de la PolimerasaRESUMEN
Construction of transgenic Anopheles mosquitoes refractory to Plasmodium requires knowledge of mosquito developmental biology. In order to study Anopheles embryology the removal or, alternatively, the permeabilization of the melanized and sclerotized egg chorion were attempted. The protocol classically used for chorion removal of Drosophila eggs was applied, with partial efficacy, to Anopheles albitarsis, a neotropical malaria vector. Each step was monitored by scanning electron microscopy and the results suggest differences in chorion composition between the two taxa. As an alternative to chorion removal, mosquito eggs were permeabilized with benserazide, an inhibitor of Dopa Decarboxylase, one of the enzymes needed for mosquito eggshell sclerotization. Embryo morphology and viability were not affected by this treatment. Permeabilization of the egg chorion allowed the ultrastructural observation of an internal homogeneous endochorion and an external compound exochorion, the latter consisting of a basal lamellar layer and protruding tubercles.
RESUMEN
The development of genetically modified vectors refractory to parasites is seen as a promising strategy in the future control of endemic diseases such as malaria. Nevertheless, knowledge of mosquito embryogenesis, a pre-requisite to the establishment of transgenic individuals, has been presently neglected. We have here studied the eggs from two neotropical malaria vectors. Eggs from Anopheles (Nyssorhynchus) albitarsis and Anopheles (Nyssorhynchus) aquasalis were analyzed by laser scanning microscopy and scanning electron microscopy and compared to those of Drosophila melanogaster. We verified basic conflicting data such as mosquito egg polarity and ultrastructure of eggshell layers. A 180 degrees rotation movement of the mosquito embryo along its longitudinal axis, a phenomenon not conserved among all Diptera, was confirmed. This early event is not taken into account by several present groups, leading to a non-consensual assignment of eggshell dorsal and ventral poles. Since embryo and egg polarities, defined during oogenesis, are the same, we propose to consider the flattened egg side as the dorsal one. The structure of Anopheles eggshell was also examined. Embryos are covered by a smooth endochorion or inner chorion layer. Outside this coat lies the compound exochorion or outer chorion layer, assembled by a thin basal lamellar layer and external tubercles. The terminology related to eggshell layers is discussed.
RESUMEN
World ecosystems differ significantly and a multidisciplinary malaria control approach must be adjusted to meet these requirements. These include a comprehensive understanding of the malaria vectors, their behavior, seasonal distribution and abundance, susceptibility to insecticides (physiological and behavioral), methods to reduce the numbers of human gametocyte carriers through effective health care systems and antimalarial drug treatment, urban malaria transmission versus rural or forest malaria transmission, and the impact of vaccine development. Many malaria vectors are members of species complexes and individual relationship to malaria transmission, seasonal distribution, bitting behavior, etc. is poorly understood. Additionaly, malaria patients are not examined for circulating gametocytes and both falciparum and vivax malaria patients may be highly infective to mosquitoes after treatment with currently used antimalarial drugs. Studies on the physiological and behavioral effects of DDT and other insecticides are inconclusive and need to be evalusted