Occurence of larval Culicidae (Diptera) in water retained in Aquascypha hydrophora (Fungus: Stereaceae) in Central Amazônia, Brazil

The community structure of insects, especially mosquito larvae, in water held in the fungus Aquascypha hydrophora (Berk.) Reid (Stereaceae) is reported. The study was done in the Adolpho Ducke Forest Reserve, 26 km east of Manaus, AM, Brazil, from September 1998 through November 1999. The most abundant entomofauna were immature Culicidae (n = 121) 91.7%, followed by adult Dytiscidae (n = 3) 2.3%, immature Chironomidae (n = 5) 3.8% and immature Tipulidae (n = 3) 2.3%. Culicidae associated with A. hydrophora comprised species of the subfamilies Anophelinae and Culicinae.

Infestation of Rhynchopsyllus pulex (Siphonaptera: Tungidae) on Molossus molossus (Chiroptera) in Southestern Brazil

The infestation of Rhynchopsyllus pulex on the bat Molossus molossus was observed using mist-nets opened between constructions in the Primatological Center, in Guapimirim, State of Rio de Janeiro, Brazil.

Derris (Lonchocarpus) urucu (Leguminosae) Extract Modifies the Peritrophic Matrix Structure of Aedes aegypti (Diptera:Culicidae)

Aqueous suspension of ethanol extracts of Derris (Lonchocarpus) urucu (Leguminosae), collected in the state of Amazonas, Brazil, were tested for larvicidal activity against the mosquito Aedes aegypti (Diptera:Culicidae). The aim of this study was to observe the alterations of peritrophic matrix in Ae. aegypti larvae treated with an aqueous suspension of D. urucu extract. Different concentrations of D. urucu root extract were tested against fourth instar larvae. One hundred percent mortality was observed at 150 µg/ml (LC50 17.6 µg/ml) 24 h following treatment. In response to D. urucu feeding, larvae excreted a large amount of amorphous feces, while control larvae did not produce feces during the assay period. Ultrastructural studies showed that larvae fed with 150 µg/ml of D. urucu extract for 4 h have an imperfect peritrophic matrix and extensive damage of the midgut epithelium. Data indicate a protective role for the peritrophic matrix. The structural modification of the peritrophic matrix is intrinsically associated with larval mortality.

Variation of the Oviposition Preferences of Aedes aegypti in Function of Substratum and Humidity

Two Aedes aegypti (L.) populations were studied in the laboratory regarding the preference for three types of breeding sites, i.e., flasks containing only water, flasks with a plant and flasks with a stick. Each of these breeding units was placed in one cage and the choice of the oviposition sites was determined for individual females and three females per experimental unit at two humidity levels. Preference for ovipositing on the water surface was observed and varied according to experimental unit and humidity. Mean hatching of eggs in water surface was 46.6%. Experiments with three females showed a more marked difference than when only one female was used. Inter and intrapopulation variability regarding oviposition sites was observed. The discrimination between the different oviposition substrates, hatching in water surface and its implication for mosquito control are discussed.

Oral Susceptibility to Yellow Fever Virus of Aedes aegypti from Brazil

The oral susceptibility to yellow fever virus was evaluated in 23 Aedes aegypti samples from Brazil. Six Ae. aegypti samples from Africa, America and Asia were also tested for comparison. Mosquito samples from Asia showed the highest infection rates. Infection rates for the Brazilian Ae. aegypti reached 48.6%, but were under 13% in 60% of sample tested. We concluded that although the low infection rates estimated for some Brazilian mosquito samples may not favor the establishment of urban cycle of yellow fever in some parts of the country, the founding of Ae. aegypti of noteworthy susceptibility to the virus in cities located in endemic and transition areas of sylvatic yellow fever, do pose a threat of the re-emergence of the urban transmission of the disease in Brazil.

Anopheles albitarsis Embryogenesis: Morphological Identification of Major Events

Anopheles albitarsis embryogenesis was analyzed through confocal microscopy of clarified eggs. Using Drosophila melanogaster as reference system, the major morphogenetic events (blastoderm, gastrulation, germ band extension, germ band retraction, dorsal closure) were identified. The kinetics of early events is proportionally similar in both systems, but late movements (from germ band retraction on) progress slower in An. albitarsis. Major differences in An. albitarsis related to D. melanogaster were: (1) pole cells do not protrude from the blastoderm; (2) the mosquito embryo undergoes a 180º rotation movement, along its longitudinal axis; (3) the head remains individualized throughout embryogenesis; (4) extraembryonary membranes surround the whole embryo. A novel kind of malaria control is under development and is based on the use of genetically modified mosquitoes. Phenotypic analysis of the embryonic development of mutants will be imposed as part of the evaluation of effectiveness and risk of employment of this strategy in the field. In order to accomplish this, knowledge of the wild type embryo is a prerequisite. Morphological studies will also serve as basis for subsequent development biology approaches.

Ochlerotatus albifasciatus in rain pools of Buenos Aires: seasonal dynamics and relation to environmental variables

The immature stages of Ochlerotatus albifasciatus develop in temporary pools. The present study aims at evaluating the seasonal dynamics of the aquatic stages of this mosquito, also analyzing the relationship among their presence and breeding success to some relevant climatic and environmental variables in the ephemeral rain pools of an urban park. Nineteen cohorts of O. albifasciatus that developed synchronously after rain events were recorded in all seasons. The proportions of mosquito-positive pools were significantly higher during the fall-winter period than in the spring-summer months (p < 0.001). The presence of this mosquito species was positively related to the amount of rain (p < 0.001), whereas negatively correlated to air temperature (p < 0.05) within a 5.2 to 29.7ºC range. The distribution of the number of cohorts per pool throughout the year was grouped (variance/mean: 3.96), indicating that these habitats were not equally suitable as breeding sites. The immature stages of O. albifasciatus were detected in pools belonging to all of the categories of surface area, depth, duration, vegetation cover, and insolation. However, the proportion of pools where immature mosquitoes were detected was positively and significantly related to surface, depth, duration, and vegetation cover. On the other hand, the proportion of mosquito-positive pools was higher at an intermediate insolation degree. Our results suggest that although preimaginal stages were present in all seasons, high temperatures may be unfavorable to larval development, and substrate vegetation may regulate water temperature. The positive relationship between the proportion of mosquito-positive pools and pool size and duration might reflect a strategy of O. albifasciatus to accomplish immature development.

Laboratory evaluation of Mesocyclops annulatus (Wierzejski, 1892) (Copepoda: Cyclopidea) as a predator of container-breeding mosquitoes in Argentina

In laboratory bioassays we tested the predatory capacity of the copepod Mesocyclops annulatus on Aedes aegypti and Culex pipiens larvae. A single adult female of M. annulatus caused 51.6% and 52.3% mortality of 50 first instar larvae of Ae. aegypti and Cx. pipiens respectively, in a 72 h test period. When alternative food was added to the containers, mortality rates declined to 16% and 10.3% for Ae. aegypti and Cx. pipiens respectively. When 50 first instar larvae of each of the two mosquito species tested were placed together with a single adult female of M. annulatus, mortality rates were 75.5% for Ae. aegypti larvae and 23.5% for Cx. pipiens larvae in a three day test period. Different density of adult females of M. annulatus ranged from 5 to 25 females produced mortality rates of Ae. aegypti first instar larvae from 50% to 100% respectively. When a single adult female of M. annulatus was exposed to an increasing number of first-instar Ae. aegypti larvae ranging from 10 to 100, 100% mortality was recorded from 1 to 25 larvae, then mortality declined to 30% with 100 larvae. The average larvae killed per 24 h period by a single copepod were 29.

A technique for preparing polytene chromosomes from Aedes aegypti (Diptera, Culicinae)

Polytene chromosome preparations were obtained from larval, pupal and adult female Malpighian tubules of Aedes aegypti. The Malpighian tubules of the pupae (0-4 h old) from larvae reared at 20ºC provided the best cytogenetic analysis. The interaction of nucleic acids and proteins that influence the spreading of the chromosomes could be reduced with the preparation technique of the sheets submitted to a stronger treatment starting with the hypotony of tissue and successive bathings with acetic acid. A simple technique should facilitate molecular cytogenetics used in the location of resistance and vector competence genes.

Larvicidal activity of the essential oil from Lippia sidoides cham. against Aedes aegypti linn

The aim of this work was to study the larvicidal activity of Lippia sidoides essential oil against Aedes aegypti larvae. The essential oil and its hydrolate (saturated solution of essential oil in water) were obtained by vapor extraction and their chemical composition determined by GL-chromatography coupled to mass spectroscopy. Bioassays were run with the essential oil, pure and diluted hydrolate and with their main constituents thymol and carvacrol. The results obtained showed that L. sidoides essential oil and its hydrolate have larvicidal action against the mosquito A. aegypti, causing an almost instantaneous mortality. Thymol, an alkylated phenol derivative and one of the major components of L. sidoides essential oil, was identified as the active principle responsible for the larvicidal action, causing 100% larval mortality at the lowest tested concentration of 0.017% (w/v). These results suggest that the essential oil of L. sidoides is promising as larvicide against A. aegypti and could be useful in the search of newer, more selective, and biodegradable larvicidal natural compounds to be used in official combat programs and at home.

Are vectors able to learn about their hosts? A case study with Aedes aegypti mosquitoes

The way in which vectors distribute themselves amongst their hosts has important epidemiological consequences. While the role played by active host choice is largely unquestioned, current knowledge relates mostly to the innate response of vectors towards stimuli signalling the presence or quality of their hosts. Many of those cues, however, can be unpredictable, and therefore prevent the incorporation of the appropriate response into the vector's behavioural repertoire unless some sort of associative learning is possible. We performed a wide range of laboratory experiments to test the learning abilities of the mosquito, Aedes aegypti. Mosquitoes were exposed to choice procedures in (1) an olfactomenter and (2) a 'visual arena'. Our goal was to determine whether the mosquitoes were able to associate unconditional stimuli (blood feeding, human breath, vibration and electrical shock) with particular odours (citral, carvone, citronella oil and eugenol) and visual patterns (horizontal or vertical black bars) to which they had been previously observed to be responsive. We found no evidence supporting the hypothesis that associative learning abilities are present in adult Ae. aegypti. We discuss the possibilities that the assays employed were either inappropriate or insufficient to detect associative learning, or that associative learning is not possible in this species.

Isoenzymatic analysis of four Anopheles (Kerteszia) bellator Dyar & Knab (Diptera: Culicidae) populations

Anopheles bellator is a small silvatic bromelia-breeding mosquito and is a primary human malaria vector species in Southern Brazil. The bromelia-breeding habitat of the species should accompany the Atlantic forest coastal distribution, where bromeliads are abundant. Nonetheless, records on An. bellator collections show a gap in the species geographical distribution. An. bellator has been recorded in Southern Brazil and in the Brazilian states of Bahia and Paraíba. It appers again in the island of Trinidad, in Trinidad and Tobago. The aim of this work was to measure gene flow between different populations of An. bellator collected in the northern and southern extremes of the geographic distribution of this species. Mosquitoes were captured in forest borders in Santa Catarina, São Paulo, and Bahia states in Brazil and in the island of Trinidad in Republic of Trinidad and Tobago. Genetic distances varied between 0.076 and 0.680, based on enzymatic profiles from 11 distinct isoenzymes. Results indicate the existence of low-level gene flow between Brazilian populations of An. bellator, and a gene flow was even lower between the Brazilian and the Trinidad populations. This finding lead us to hypothesize that An. bellator did not spread along the coast, but reached northeastern areas through inland routes.

Laboratory evaluation of methanolic extract of Atlantia monophylla (Family: Rutaceae) against immature stages of mosquitoes and non-target organisms

Methanolic extracts of the leaves of Atlantia monophylla (Rutaceae) were evaluated for mosquitocidal activity against immature stages of three mosquito species, Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti in the laboratory.Larvae of Cx. quinquefasciatus and pupae of An. stephensi were found more susceptible, with LC50 values of 0.14 mg/l and 0.05 mg/l, respectively. Insect growth regulating activity of this extract was more pronounced against Ae. aegypti, with EI50 value 0.002 mg/l. The extract was found safe to aquatic mosquito predators Gambusia affinis, Poecilia reticulata, and Diplonychus indicus, with the respective LC50 values of 23.4, 21.3, and 5.7 mg/l. The results indicate that the mosquitocidal effects of the extract of this plant were comparable to neem extract and certain synthetic chemical larvicides like fenthion, methoprene, etc.

Seasonal pattern of abundance of Aedes aegypti (Diptera: Culicidae) in Buenos Aires city, Argentina

In Buenos Aires, the most crowded city of Argentina, there is a potential risk of dengue virus transmission by the mosquito Aedes aegypti during late summer. The temporal patterns of oviposition activity and abundance of breeding sites of this vector were studied in two cemeteries of the city. Between September 1998 and August 1999, we examined 142 ovitraps weekly and a total of 18,010 water-filled containers. Both study areas showed remarkable differences in the percentages of positive ovitraps (19% vs 8%) and breeding sites (18% vs 1%), but similar temporal abundance patterns. The percentage of breeding sites was higher in summer and autumn than in spring and winter, and the percentage of positive ovitraps was higher in summer than in the other three seasons. Immatures were recorded from the first week of October to the second week of July, and oviposition activity from the third week of October until the end of April. In both cemeteries and with both methodologies the highest infestation levels were registered in March (ovitraps: 41.8% and 20.6%, breeding sites: 39.2% and 3.4%). These highest abundances took place after several months with mean temperatures above 20ºC and accumulated rainfalls above 150 mm. A sharp decline in oviposition activity was observed when monthly mean temperature decreased to 16.5ºC, and no eggs were found below 14.8ºC. Seasonal fluctuation of Ae. aegypti abundances in mid-latitudes like Buenos Aires would allow reduction of the egg mosquito population through the elimination of containers during the coldest months, which are free of adults.

Isoenzimatic analysis of four Anopheles (Kerteszia) cruzii (Diptera: Culicidae) populations of Brazil

Anopheles cruzii is a small sylvatic mosquito and primary human Plasmodium vector in Southern Brazil. The distribution of this bromeliad-breeding mosquito follows the Atlantic forest coastal distribution, where bromeliads are abundant. Morphological, genetic, and molecular polymorphisms among different populations have been reported and it has recently been suggested that An. cruzii is a complex of cryptic species. The aim of this work is to analyze the gene flow between different populations of An. cruzii collected in four localities within the geographic distribution range of the species, and to examine if An. cruzii is a complex of cryptic species. The genetic distances show that populations of the states of Santa Catarina, São Paulo, and Rio de Janeiro are genetically closer (0.032 to 0.083) than populations of Bahia (0.364 to 0.853) based on profiles from 10 distinct isoenzyme loci. The Fst was lower (0.077) when the Bahia population was excluded than when it was included (0.300) in the analyses. The inferred number of migrants per generation was 2.99 individuals among populations from the states of Santa Catarina, São Paulo, and Rio de Janeiro and 0.58 migrants per generation among all populations. Results suggest that An. cruzii is a complex of species and that the specimes of state of Bahia can be considered as belonging to a species that is distinct from other three closely-related populations studied.