Rafaela Bruno - Academia.edu (original) (raw)

Papers by Rafaela Bruno

Research paper thumbnail of Everybody loves sugar: first report of plant feeding in triatomines

Background: Triatomines, which are the vectors of Trypanosoma cruzi, have been considered to be e... more Background: Triatomines, which are the vectors of Trypanosoma cruzi, have been considered to be exclusive blood feeders for more than 100 years, since the discovery of Chagas disease.

Research paper thumbnail of DNA Barcoding of Neotropical Sand Flies (Diptera, Psychodidae, Phlebotominae): Species Identification and Discovery within Brazil

PLOS ONE, 2015

DNA barcoding has been an effective tool for species identification in several animal groups. Her... more DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23-19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.

Research paper thumbnail of Clocks do not tick in unison: isolation of Clock and vrille shed new light on the clockwork model of the sand fly Lutzomyia longipalpis

Parasites & Vectors, 2015

Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission ... more Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission to humans. The sand fly Lutzomyia longipalpis is the main vector of visceral leishmaniasis in America and concentrates its activity around dusk. Despite the accumulation of behavioral data, very little is known about the molecular bases of the clock mechanism in this species. This study aims to characterize, within an evolutionary perspective, two important circadian clock genes, Clock and vrille. We have cloned and isolated the coding sequence of L. longipalpis' genes Clock and vrille. The former is structured in eight exons and encodes a protein of 696 amino acids, and the latter comprises three exons and translates to a protein of 469 amino acids. When compared to other insects' orthologues, L. longipalpis CLOCK shows a high degree of conservation in the functional domains bHLH and PAS, but a much shorter glutamine-rich (poly-Q) C-terminal region. As for L. longipalpis VRILLE, a high degree of conservation was found in the bZIP domain. To support these observations and provide an elegant view of the evolution of both genes in insects, phylogenetic analyses based on maximum-likelihood and Bayesian inferences were performed, corroborating the previously known insect systematics. The isolation and phylogenetic analyses of Clock and vrille orthologues in L. longipalpis bring novel and important data to characterize this species' circadian clock. Interestingly, the poly-Q shortening observed in CLOCK suggests that its transcription activity might be impaired and we speculate if this effect could be compensated by other clock factors such as CYCLE.

Research paper thumbnail of Phenotypic differentiation in love song traits among sibling species of the Lutzomyia longipalpis complex in Brazil

Parasites & vectors, 2015

Brazilian populations of Lutzomyia longipalpis may constitute a complex of cryptic species, and t... more Brazilian populations of Lutzomyia longipalpis may constitute a complex of cryptic species, and this report investigates the distribution and number of potential sibling species. One of the main differences observed among Brazilian populations is the type of acoustic signal produced by males during copulation. These copulation song differences seem to be evolving faster than neutral molecular markers and have been suggested to contribute to insemination failure observed in crosses between these sibling species. In previous studies, two main types of copulation songs were found, burst-type and pulse-type. The latter type can, in turn, be further subdivided into five different patterns. We recorded male song from 13 new populations of the L. longipalpis complex from Brazil and compared the songs with 12 already available. Out of these 25 populations, 16 produce burst-type and 9 produce pulse-type songs. We performed a principal component analysis in these two main groups separately an...

Research paper thumbnail of Fine Structure of the Male Reproductive System and Reproductive Behavior of Lutzomyia longipalpis Sandflies (Diptera: Psychodidae: Phlebotominae)

PLoS ONE, 2013

Background: The male reproductive system of insects can have several tissues responsible for the ... more Background: The male reproductive system of insects can have several tissues responsible for the secretion of seminal fluid proteins (SFPs), such as testes, accessory glands, seminal vesicles, ejaculatory duct and ejaculatory bulb. The SFPs are transferred during mating and can induce several physiological and behavioral changes in females, such as increase in oviposition and decrease in sexual receptivity after copulation. The phlebotomine Lutzomyia longipalpis is the main vector of visceral leishmaniasis. Despite its medical importance, little is known about its reproductive biology. Here we present morphological aspects of the male L. longipalpis reproductive system by light, scanning and transmission electron microscopy, and compare the mating frequency of both virgin and previously mated females.

Research paper thumbnail of Potential impact of a presumed increase in the biting activity of dengue-virus-infected Aedes aegypti (Diptera: Culicidae) females on virus transmission dynamics

Memórias do Instituto Oswaldo Cruz, 2011

Recently, we showed that infection with dengue virus increases the locomotor activity of Aedes ae... more Recently, we showed that infection with dengue virus increases the locomotor activity of Aedes aegypti females. We speculate that the observed increased locomotor activity could potentially increase the chances of finding a suitable host and, as a consequence, the relative biting rate of infected mosquitoes. We used a mathematical model to investigate the impact of the increased locomotor activity by assuming that this activity translated into an increased biting rate for infected mosquitoes. The results show that the increased biting rate resulted in dengue outbreaks with greater numbers of primary and secondary infections and with more severe biennial epidemics.

Research paper thumbnail of Effects of insemination and blood-feeding on locomotor activity of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) females under laboratory conditions

Parasites & Vectors, 2014

Dengue is an arbovirus disease transmitted by two Aedes mosquitoes: Ae. aegypti and Ae. albopictu... more Dengue is an arbovirus disease transmitted by two Aedes mosquitoes: Ae. aegypti and Ae. albopictus. Virgin females of these two species generally show a bimodal and diurnal pattern of activity, with early morning and late afternoon peaks. Although some studies on the flight activity of virgin, inseminated and blood-fed Ae. aegypti females have been carried out under laboratory conditions, little is known about the effects of such physiological states on the locomotor activity of Ae. albopictus and Ae. aegypti females. The aim of this study was to analyze, under laboratory conditions, the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under LD 12:12, at 25°C. Both Ae. albopictus and Ae. aegypti females were obtained from established laboratory colonies. Control groups were represented by virgin/unfed Ae. albopictus and Ae. aegypti females. Experiments were conducted under laboratory conditions, using an activity monitor that registers individual activity every thirty minutes. Virgin/unfed Ae. albopictus and Ae. aegypti females showed a diurnal and bimodal pattern of locomotor activity, with peaks at early morning and late afternoon. Insemination and blood-feeding significantly decreased the locomotor activity of Ae. aegypti females, but inseminated/blood-fed Ae. aegypti and Ae. albopictus females showed a similar significant decrease on the locomotor activity compared to virgin/unfed females. This study is the first demonstration of the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under artificial conditions. Data suggest that Ae. albopictus and Ae. aegypti females respond in different ways to physiological status changes and such divergence between these two dengue vectors, associated with several ecological differences, could be related to the greater dengue vectorial capacity of Ae. aegypti in Americas in comparison to Ae. albopictus.

Research paper thumbnail of Circadian clock of Aedes aegypti: effects of blood-feeding, insemination and RNA interference

Memórias do Instituto Oswaldo Cruz, 2013

Mosquitoes are the culprits of some of the most important vector borne diseases. A species' poten... more Mosquitoes are the culprits of some of the most important vector borne diseases. A species' potential as a vector is directly dependent on their pattern of behaviour, which is known to change according to the female's physiological status such as whether the female is virgin/mated and unfed/blood-fed. However, the molecular mechanism triggered by and/or responsible for such modulations in behaviour is poorly understood. Clock genes are known to be responsible for the control of circadian behaviour in several species. Here we investigate the impact mating and blood-feeding have upon the expression of these genes in the mosquito Aedes aegypti. We show that blood intake, but not insemination, is responsible for the down-regulation of clock genes. Using RNA interference, we observe a slight reduction in the evening activity peak in the fourth day after dstim injection. These data suggest that, as in Drosophila, clock gene expression, circadian behaviour and environmental light regimens are interconnected in Ae. aegypti.

Research paper thumbnail of Male accessory gland substances from Aedes albopictus affect the locomotor activity of Aedes aegypti females

Memórias do Instituto Oswaldo Cruz, 2013

Dengue is one of the world&am... more Dengue is one of the world's most important mosquito-borne diseases and is usually transmitted by one of two vector species: Aedes aegypti or Aedes albopictus . These two diurnal mosquitoes are frequently found coexisting in similar habitats, enabling interactions between adults, such as cross-mating. The objective of this study was to assess cross-mating between Ae. aegypti females and Ae. albopictus males under artificial conditions and evaluate the locomotor activity of Ae. aegypti virgin females injected with male accessory gland (MAG) homogenates to infer the physiological and behavioural responses to interspecific mating. After seven days of exposure, 3.3-16% of Ae. aegypti females mated with Ae. albopictus males. Virgin Ae. aegypti females injected with conspecific and heterospecific MAGs showed a general decrease in locomotor activity compared to controls and were refractory to mating with conspecific males. The reduction in diurnal locomotor activity induced by injections of conspecific or heterospecific MAGs is consistent with regulation of female reproductive activities by male substances, which are capable of sterilising female Ae. aegypti through satyrisation by Ae. albopictus.

Research paper thumbnail of Effects of temperature and photoperiod on daily activity rhythms of Lutzomyia longipalpis (Diptera: Psychodidae)

Parasites & Vectors, 2014

Background: Insect vectors have been established as models in Chronobiology for many decades, and... more Background: Insect vectors have been established as models in Chronobiology for many decades, and recent studies have demonstrated a close relationship between the circadian clock machinery, daily rhythms of activity and vectorial capacity. Lutzomyia longipalpis, the primary vector of Leishmania (Leishmania) infantum in the New World, is reported to have crepuscular/nocturnal activity in the wild. However, most of these studies applied hourly CDC trap captures, which is a good indicative of L. longipalpis behaviour, but has limited accuracy due to the inability to record the daily activity of a single insect during consecutive days. In addition, very little is known about the activity pattern of L. longipalpis under seasonal variations of average temperature and day length in controlled laboratory conditions. Methods: We recorded the locomotor activity of L. longipalpis males under different artificial regimes of temperature and photoperiod. First, in order to test the effects of temperature on the activity, sandflies were submitted to regimes of light/dark cycles similar to the equinox photoperiod (LD 12:12) combined with different constant temperatures (20°C, 25°C and 30°C). In addition, we recorded sandfly locomotor activity under a mild constant temperature (25°C with different day length regimes: 8 hours, 12 hours and 16 hours). Results: L. longipalpis exhibited more activity at night, initiating dusk-related activity (onset time) at higher rather than lower temperatures. In parallel, changes of photoperiod affected anticipation as well as all the patterns of activity (onset, peak and offset time). However, under LD 16:08, sandflies presented the earliest values of maximum peak and offset times, contrary to other regimes. Conclusions: Herein, we showed that light and temperature modulate L. longipalpis behaviour under controlled laboratory conditions, suggesting that sandflies might use environmental information to sustain their crepuscular/ nocturnal activity, as well as other important aspects as mating and host-seeking at appropriate times in different seasons. Our results depict previously unappreciated aspects of the L. longipalpis daily rhythms of activity that might have important epidemiological implications.

Research paper thumbnail of Evidence for gene duplication in the voltage-gated sodium channel gene of Aedes aegypti

Evolution, Medicine, and Public Health, 2013

Background and objectives: Mutations in the voltage-gated sodium channel gene (Na V ), known as k... more Background and objectives: Mutations in the voltage-gated sodium channel gene (Na V ), known as kdr mutations, are associated with pyrethroid and DDT insecticide resistance in a number of species. In the mosquito dengue vector Aedes aegypti, besides kdr, other polymorphisms allowed grouping AaNa V sequences as type 'A' or 'B'. Here, we point a series of evidences that these polymorphisms are actually involved in a gene duplication event.

Research paper thumbnail of Dengue Infection Increases the Locomotor Activity of Aedes aegypti Females

PLoS ONE, 2011

Background: Aedes aegypti is the main vector of the virus causing Dengue fever, a disease that ha... more Background: Aedes aegypti is the main vector of the virus causing Dengue fever, a disease that has increased dramatically in importance in recent decades, affecting many tropical and sub-tropical areas of the globe. It is known that viruses and other parasites can potentially alter vector behavior. We investigated whether infection with Dengue virus modifies the behavior of Aedes aegypti females with respect to their activity level.

Research paper thumbnail of Multilocus Analysis of Divergence and Introgression in Sympatric and Allopatric Sibling Species of the Lutzomyia longipalpis Complex in Brazil

PLoS Neglected Tropical Diseases, 2013

Background: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in Latin America, is... more Background: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in Latin America, is a complex of sibling species. In Brazil, a number of very closely related sibling species have been revealed by the analyses of copulation songs, sex pheromones and molecular markers. However, the level of divergence and gene flow between the sibling species remains unclear. Brazilian populations of this vector can be divided in two main groups: one producing Burst-type songs and the Cembrene-1 pheromone and a second more diverse group producing various Pulse song subtypes and different pheromones.

Research paper thumbnail of Distribution and dissemination of the Val1016Ile and Phe1534Cys Kdr mutations in Aedes aegypti Brazilian natural populations

Parasites & Vectors, 2014

Background: The chemical control of the mosquito Aedes aegypti, the major vector of dengue, is be... more Background: The chemical control of the mosquito Aedes aegypti, the major vector of dengue, is being seriously threatened due to the development of pyrethroid resistance. Substitutions in the 1016 and 1534 sites of the voltage gated sodium channel (AaNa V ), commonly known as kdr mutations, confer the mosquito with knockdown resistance. Our aim was to evaluate the allelic composition of natural populations of Brazilian Ae. aegypti at both kdr sites.

Research paper thumbnail of Why do we need alternative tools to control mosquito-borne diseases in Latin America?

Memórias do Instituto Oswaldo Cruz, 2012

In this opinion paper, we discuss the potential and challenges of using the symbiont Wolbachia to... more In this opinion paper, we discuss the potential and challenges of using the symbiont Wolbachia to block mosquito transmitted diseases such as dengue, malaria and chikungunya in Latin America.

Research paper thumbnail of Analysis of cycle Gene Expression in Aedes aegypti Brains by In Situ Hybridization

Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors... more Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors, relatively little is known about the molecular mechanisms underlying processes involved in the temporal pattern of its activity and host seeking behavior. In this study, we analyzed the expression of the cycle (cyc) gene, one of the core components of the circadian clock, in Ae. aegypti brains by in situ hybridization at two different time points in light-dark conditions and compared the results with those obtained using a quantitative PCR assay (qPCR). Within the brain, differential labeling was detected according to distinct areas empirically pre-defined. Six out of seven of these areas showed significantly higher staining at ZT3 (three hours after light-on) compared to ZT11 (one before light-off), which is consistent with the qPCR data. Predominant staining was observed in three of those areas which correspond to positions of the optical and antennal lobes, as well as the region where the neurons controlling activity rhythms are presumably localized.

Research paper thumbnail of Alexandre Afranio Peixoto (1963-2013)

Genetics and Molecular Biology, 2013

Research paper thumbnail of Everybody loves sugar: first report of plant feeding in triatomines

Background: Triatomines, which are the vectors of Trypanosoma cruzi, have been considered to be e... more Background: Triatomines, which are the vectors of Trypanosoma cruzi, have been considered to be exclusive blood feeders for more than 100 years, since the discovery of Chagas disease.

Research paper thumbnail of DNA Barcoding of Neotropical Sand Flies (Diptera, Psychodidae, Phlebotominae): Species Identification and Discovery within Brazil

PLOS ONE, 2015

DNA barcoding has been an effective tool for species identification in several animal groups. Her... more DNA barcoding has been an effective tool for species identification in several animal groups. Here, we used DNA barcoding to discriminate between 47 morphologically distinct species of Brazilian sand flies. DNA barcodes correctly identified approximately 90% of the sampled taxa (42 morphologically distinct species) using clustering based on neighbor-joining distance, of which four species showed comparatively higher maximum values of divergence (range 4.23-19.04%), indicating cryptic diversity. The DNA barcodes also corroborated the resurrection of two species within the shannoni complex and provided an efficient tool to differentiate between morphologically indistinguishable females of closely related species. Taken together, our results validate the effectiveness of DNA barcoding for species identification and the discovery of cryptic diversity in sand flies from Brazil.

Research paper thumbnail of Clocks do not tick in unison: isolation of Clock and vrille shed new light on the clockwork model of the sand fly Lutzomyia longipalpis

Parasites & Vectors, 2015

Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission ... more Behavior rhythms of insect vectors directly interfere with the dynamics of pathogen transmission to humans. The sand fly Lutzomyia longipalpis is the main vector of visceral leishmaniasis in America and concentrates its activity around dusk. Despite the accumulation of behavioral data, very little is known about the molecular bases of the clock mechanism in this species. This study aims to characterize, within an evolutionary perspective, two important circadian clock genes, Clock and vrille. We have cloned and isolated the coding sequence of L. longipalpis' genes Clock and vrille. The former is structured in eight exons and encodes a protein of 696 amino acids, and the latter comprises three exons and translates to a protein of 469 amino acids. When compared to other insects' orthologues, L. longipalpis CLOCK shows a high degree of conservation in the functional domains bHLH and PAS, but a much shorter glutamine-rich (poly-Q) C-terminal region. As for L. longipalpis VRILLE, a high degree of conservation was found in the bZIP domain. To support these observations and provide an elegant view of the evolution of both genes in insects, phylogenetic analyses based on maximum-likelihood and Bayesian inferences were performed, corroborating the previously known insect systematics. The isolation and phylogenetic analyses of Clock and vrille orthologues in L. longipalpis bring novel and important data to characterize this species' circadian clock. Interestingly, the poly-Q shortening observed in CLOCK suggests that its transcription activity might be impaired and we speculate if this effect could be compensated by other clock factors such as CYCLE.

Research paper thumbnail of Phenotypic differentiation in love song traits among sibling species of the Lutzomyia longipalpis complex in Brazil

Parasites & vectors, 2015

Brazilian populations of Lutzomyia longipalpis may constitute a complex of cryptic species, and t... more Brazilian populations of Lutzomyia longipalpis may constitute a complex of cryptic species, and this report investigates the distribution and number of potential sibling species. One of the main differences observed among Brazilian populations is the type of acoustic signal produced by males during copulation. These copulation song differences seem to be evolving faster than neutral molecular markers and have been suggested to contribute to insemination failure observed in crosses between these sibling species. In previous studies, two main types of copulation songs were found, burst-type and pulse-type. The latter type can, in turn, be further subdivided into five different patterns. We recorded male song from 13 new populations of the L. longipalpis complex from Brazil and compared the songs with 12 already available. Out of these 25 populations, 16 produce burst-type and 9 produce pulse-type songs. We performed a principal component analysis in these two main groups separately an...

Research paper thumbnail of Fine Structure of the Male Reproductive System and Reproductive Behavior of Lutzomyia longipalpis Sandflies (Diptera: Psychodidae: Phlebotominae)

PLoS ONE, 2013

Background: The male reproductive system of insects can have several tissues responsible for the ... more Background: The male reproductive system of insects can have several tissues responsible for the secretion of seminal fluid proteins (SFPs), such as testes, accessory glands, seminal vesicles, ejaculatory duct and ejaculatory bulb. The SFPs are transferred during mating and can induce several physiological and behavioral changes in females, such as increase in oviposition and decrease in sexual receptivity after copulation. The phlebotomine Lutzomyia longipalpis is the main vector of visceral leishmaniasis. Despite its medical importance, little is known about its reproductive biology. Here we present morphological aspects of the male L. longipalpis reproductive system by light, scanning and transmission electron microscopy, and compare the mating frequency of both virgin and previously mated females.

Research paper thumbnail of Potential impact of a presumed increase in the biting activity of dengue-virus-infected Aedes aegypti (Diptera: Culicidae) females on virus transmission dynamics

Memórias do Instituto Oswaldo Cruz, 2011

Recently, we showed that infection with dengue virus increases the locomotor activity of Aedes ae... more Recently, we showed that infection with dengue virus increases the locomotor activity of Aedes aegypti females. We speculate that the observed increased locomotor activity could potentially increase the chances of finding a suitable host and, as a consequence, the relative biting rate of infected mosquitoes. We used a mathematical model to investigate the impact of the increased locomotor activity by assuming that this activity translated into an increased biting rate for infected mosquitoes. The results show that the increased biting rate resulted in dengue outbreaks with greater numbers of primary and secondary infections and with more severe biennial epidemics.

Research paper thumbnail of Effects of insemination and blood-feeding on locomotor activity of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) females under laboratory conditions

Parasites & Vectors, 2014

Dengue is an arbovirus disease transmitted by two Aedes mosquitoes: Ae. aegypti and Ae. albopictu... more Dengue is an arbovirus disease transmitted by two Aedes mosquitoes: Ae. aegypti and Ae. albopictus. Virgin females of these two species generally show a bimodal and diurnal pattern of activity, with early morning and late afternoon peaks. Although some studies on the flight activity of virgin, inseminated and blood-fed Ae. aegypti females have been carried out under laboratory conditions, little is known about the effects of such physiological states on the locomotor activity of Ae. albopictus and Ae. aegypti females. The aim of this study was to analyze, under laboratory conditions, the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under LD 12:12, at 25°C. Both Ae. albopictus and Ae. aegypti females were obtained from established laboratory colonies. Control groups were represented by virgin/unfed Ae. albopictus and Ae. aegypti females. Experiments were conducted under laboratory conditions, using an activity monitor that registers individual activity every thirty minutes. Virgin/unfed Ae. albopictus and Ae. aegypti females showed a diurnal and bimodal pattern of locomotor activity, with peaks at early morning and late afternoon. Insemination and blood-feeding significantly decreased the locomotor activity of Ae. aegypti females, but inseminated/blood-fed Ae. aegypti and Ae. albopictus females showed a similar significant decrease on the locomotor activity compared to virgin/unfed females. This study is the first demonstration of the effects of insemination and blood-feeding on the locomotor activity of Ae. albopictus and Ae. aegypti females under artificial conditions. Data suggest that Ae. albopictus and Ae. aegypti females respond in different ways to physiological status changes and such divergence between these two dengue vectors, associated with several ecological differences, could be related to the greater dengue vectorial capacity of Ae. aegypti in Americas in comparison to Ae. albopictus.

Research paper thumbnail of Circadian clock of Aedes aegypti: effects of blood-feeding, insemination and RNA interference

Memórias do Instituto Oswaldo Cruz, 2013

Mosquitoes are the culprits of some of the most important vector borne diseases. A species' poten... more Mosquitoes are the culprits of some of the most important vector borne diseases. A species' potential as a vector is directly dependent on their pattern of behaviour, which is known to change according to the female's physiological status such as whether the female is virgin/mated and unfed/blood-fed. However, the molecular mechanism triggered by and/or responsible for such modulations in behaviour is poorly understood. Clock genes are known to be responsible for the control of circadian behaviour in several species. Here we investigate the impact mating and blood-feeding have upon the expression of these genes in the mosquito Aedes aegypti. We show that blood intake, but not insemination, is responsible for the down-regulation of clock genes. Using RNA interference, we observe a slight reduction in the evening activity peak in the fourth day after dstim injection. These data suggest that, as in Drosophila, clock gene expression, circadian behaviour and environmental light regimens are interconnected in Ae. aegypti.

Research paper thumbnail of Male accessory gland substances from Aedes albopictus affect the locomotor activity of Aedes aegypti females

Memórias do Instituto Oswaldo Cruz, 2013

Dengue is one of the world&am... more Dengue is one of the world's most important mosquito-borne diseases and is usually transmitted by one of two vector species: Aedes aegypti or Aedes albopictus . These two diurnal mosquitoes are frequently found coexisting in similar habitats, enabling interactions between adults, such as cross-mating. The objective of this study was to assess cross-mating between Ae. aegypti females and Ae. albopictus males under artificial conditions and evaluate the locomotor activity of Ae. aegypti virgin females injected with male accessory gland (MAG) homogenates to infer the physiological and behavioural responses to interspecific mating. After seven days of exposure, 3.3-16% of Ae. aegypti females mated with Ae. albopictus males. Virgin Ae. aegypti females injected with conspecific and heterospecific MAGs showed a general decrease in locomotor activity compared to controls and were refractory to mating with conspecific males. The reduction in diurnal locomotor activity induced by injections of conspecific or heterospecific MAGs is consistent with regulation of female reproductive activities by male substances, which are capable of sterilising female Ae. aegypti through satyrisation by Ae. albopictus.

Research paper thumbnail of Effects of temperature and photoperiod on daily activity rhythms of Lutzomyia longipalpis (Diptera: Psychodidae)

Parasites & Vectors, 2014

Background: Insect vectors have been established as models in Chronobiology for many decades, and... more Background: Insect vectors have been established as models in Chronobiology for many decades, and recent studies have demonstrated a close relationship between the circadian clock machinery, daily rhythms of activity and vectorial capacity. Lutzomyia longipalpis, the primary vector of Leishmania (Leishmania) infantum in the New World, is reported to have crepuscular/nocturnal activity in the wild. However, most of these studies applied hourly CDC trap captures, which is a good indicative of L. longipalpis behaviour, but has limited accuracy due to the inability to record the daily activity of a single insect during consecutive days. In addition, very little is known about the activity pattern of L. longipalpis under seasonal variations of average temperature and day length in controlled laboratory conditions. Methods: We recorded the locomotor activity of L. longipalpis males under different artificial regimes of temperature and photoperiod. First, in order to test the effects of temperature on the activity, sandflies were submitted to regimes of light/dark cycles similar to the equinox photoperiod (LD 12:12) combined with different constant temperatures (20°C, 25°C and 30°C). In addition, we recorded sandfly locomotor activity under a mild constant temperature (25°C with different day length regimes: 8 hours, 12 hours and 16 hours). Results: L. longipalpis exhibited more activity at night, initiating dusk-related activity (onset time) at higher rather than lower temperatures. In parallel, changes of photoperiod affected anticipation as well as all the patterns of activity (onset, peak and offset time). However, under LD 16:08, sandflies presented the earliest values of maximum peak and offset times, contrary to other regimes. Conclusions: Herein, we showed that light and temperature modulate L. longipalpis behaviour under controlled laboratory conditions, suggesting that sandflies might use environmental information to sustain their crepuscular/ nocturnal activity, as well as other important aspects as mating and host-seeking at appropriate times in different seasons. Our results depict previously unappreciated aspects of the L. longipalpis daily rhythms of activity that might have important epidemiological implications.

Research paper thumbnail of Evidence for gene duplication in the voltage-gated sodium channel gene of Aedes aegypti

Evolution, Medicine, and Public Health, 2013

Background and objectives: Mutations in the voltage-gated sodium channel gene (Na V ), known as k... more Background and objectives: Mutations in the voltage-gated sodium channel gene (Na V ), known as kdr mutations, are associated with pyrethroid and DDT insecticide resistance in a number of species. In the mosquito dengue vector Aedes aegypti, besides kdr, other polymorphisms allowed grouping AaNa V sequences as type 'A' or 'B'. Here, we point a series of evidences that these polymorphisms are actually involved in a gene duplication event.

Research paper thumbnail of Dengue Infection Increases the Locomotor Activity of Aedes aegypti Females

PLoS ONE, 2011

Background: Aedes aegypti is the main vector of the virus causing Dengue fever, a disease that ha... more Background: Aedes aegypti is the main vector of the virus causing Dengue fever, a disease that has increased dramatically in importance in recent decades, affecting many tropical and sub-tropical areas of the globe. It is known that viruses and other parasites can potentially alter vector behavior. We investigated whether infection with Dengue virus modifies the behavior of Aedes aegypti females with respect to their activity level.

Research paper thumbnail of Multilocus Analysis of Divergence and Introgression in Sympatric and Allopatric Sibling Species of the Lutzomyia longipalpis Complex in Brazil

PLoS Neglected Tropical Diseases, 2013

Background: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in Latin America, is... more Background: Lutzomyia longipalpis, the main vector of visceral leishmaniasis in Latin America, is a complex of sibling species. In Brazil, a number of very closely related sibling species have been revealed by the analyses of copulation songs, sex pheromones and molecular markers. However, the level of divergence and gene flow between the sibling species remains unclear. Brazilian populations of this vector can be divided in two main groups: one producing Burst-type songs and the Cembrene-1 pheromone and a second more diverse group producing various Pulse song subtypes and different pheromones.

Research paper thumbnail of Distribution and dissemination of the Val1016Ile and Phe1534Cys Kdr mutations in Aedes aegypti Brazilian natural populations

Parasites & Vectors, 2014

Background: The chemical control of the mosquito Aedes aegypti, the major vector of dengue, is be... more Background: The chemical control of the mosquito Aedes aegypti, the major vector of dengue, is being seriously threatened due to the development of pyrethroid resistance. Substitutions in the 1016 and 1534 sites of the voltage gated sodium channel (AaNa V ), commonly known as kdr mutations, confer the mosquito with knockdown resistance. Our aim was to evaluate the allelic composition of natural populations of Brazilian Ae. aegypti at both kdr sites.

Research paper thumbnail of Why do we need alternative tools to control mosquito-borne diseases in Latin America?

Memórias do Instituto Oswaldo Cruz, 2012

In this opinion paper, we discuss the potential and challenges of using the symbiont Wolbachia to... more In this opinion paper, we discuss the potential and challenges of using the symbiont Wolbachia to block mosquito transmitted diseases such as dengue, malaria and chikungunya in Latin America.

Research paper thumbnail of Analysis of cycle Gene Expression in Aedes aegypti Brains by In Situ Hybridization

Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors... more Even though the blood-sucking mosquito Aedes aegypti is one of the most important disease vectors, relatively little is known about the molecular mechanisms underlying processes involved in the temporal pattern of its activity and host seeking behavior. In this study, we analyzed the expression of the cycle (cyc) gene, one of the core components of the circadian clock, in Ae. aegypti brains by in situ hybridization at two different time points in light-dark conditions and compared the results with those obtained using a quantitative PCR assay (qPCR). Within the brain, differential labeling was detected according to distinct areas empirically pre-defined. Six out of seven of these areas showed significantly higher staining at ZT3 (three hours after light-on) compared to ZT11 (one before light-off), which is consistent with the qPCR data. Predominant staining was observed in three of those areas which correspond to positions of the optical and antennal lobes, as well as the region where the neurons controlling activity rhythms are presumably localized.

Research paper thumbnail of Alexandre Afranio Peixoto (1963-2013)

Genetics and Molecular Biology, 2013