Rapid Loss of Resistance to Satyrization in Invasive Mosquitoes and the Effects of Age on Interspecific Mating Frequency.

In several areas where Aedes aegypti (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) have come in contact following successful invasions, Ae. aegypti have been rapidly displaced by Ae. albopictus. Recent work has confirmed that mating interference, in the form of satyrization, is likely a driving factor in these competitive displacements. However, in sites of sympatry, Ae. aegypti females evolve resistance to satyrization, and in the laboratory, satyrization-susceptible Ae. aegypti can evolve resistance within a few generations of cage exposure to Ae. albopictus. The resistance trait may be costly to maintain, as satyrization-resistant females have been shown to invest more time in mate selection. Here, we show that increased satyrization-resistance does not persist in the absence of satyrization pressure, confirming a cost to maintaining this trait. Ae. aegypti lines that had been previously selected for increased satyrization-resistance demonstrated rapid decreases in this trait over eight generations. Support for the hypothesis that condition-dependent mate preferences lead to variation in mating choice among individuals of differing quality within a population has been accumulating. Here we examine how age-related changes in reproductive effort or reproductive value may result in age-dependent mate choice. We hypothesize that older, lower quality individuals may be more likely to accept a subpar mating, in this case an interspecific mating. Our results demonstrate that mosquito age significantly affects interspecific mating rates with older mosquitoes (male and female) engaging in interspecific mating more frequently than younger counterparts (26.32 vs 9.41%). We discuss the possibility of age-related signal variation causing the breakdown of mating barriers with age.

Correction to: Socioeconomic and demographic characterization of an endemic malaria region in Brazil by multiple correspondence analysis.

After publication of the article [1], it has been brought to our attention that the y-axis of Fig. 6 has been labeled incorrectly. It should read "linear predictor". This has now been corrected in the original article.

Socioeconomic and demographic characterization of an endemic malaria region in Brazil by multiple correspondence analysis.

BACKGROUND: In the process of geographical retraction of malaria, some important endemicity pockets remain. Here, we report results from a study developed to obtain detailed community data from an important malaria hotspot in Latin America (Alto Juruá, Acre, Brazil), to investigate the association of malaria with socioeconomic, demographic and living conditions. METHODS: A household survey was conducted in 40 localities (n = 520) of Mâncio Lima and Rodrigues Alves municipalities, Acre state. Information on previous malaria, schooling, age, gender, income, occupation, household structure, habits and behaviors related to malaria exposure was collected. Multiple correspondence analysis (MCA) was applied to characterize similarities between households and identify gradients. The association of these gradients with malaria was assessed using regression. RESULTS: The first three dimensions of MCA accounted for almost 50% of the variability between households. The first dimension defined an urban/rurality gradient, where urbanization was associated with the presence of roads, basic services as garbage collection, water treatment, power grid energy, and less contact with the forest. There is a significant association between this axis and the probability of malaria at the household level, OR = 1.92 (1.23-3.02). The second dimension described a gradient from rural settlements in agricultural areas to those in forested areas. Access via dirt road or river, access to electricity power-grid services and aquaculture were important variables. Malaria was at lower risk at the forested area, OR = 0.55 (1.23-1.12). The third axis detected intraurban differences and did not correlate with malaria. CONCLUSIONS: Living conditions in the study area are strongly geographically structured. Although malaria is found throughout all the landscapes, household traits can explain part of the variation found in the odds of having malaria. It is expected these results stimulate further discussions on modelling approaches targeting a more systemic and multi-level view of malaria dynamics.

Dengue serotype circulation in natural populations of Aedes aegypti.

Ae. aegypti is the main vector of dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viruses. The transmission dynamics of these arboviruses, especially the arboviral circulation in the mosquito population during low and high transmission seasons in endemic areas are still poorly understood. We conducted an entomological survey to determine dengue infection rates in Ae. aegypti and Aedes albopictus. These collections were performed in 2012-2013 during a Rio de Janeiro epidemic, just before the introduction and spread of ZIKV and CHIKV in the city. MosquiTrap© and BG-Sentinel traps were installed in three fixed and seven itinerant neighborhoods each month over ten months. Mosquitoes were in supernatants pools tested and individually confirmed for DENV infection using RT-PCR. A total of 3053 Aedes mosquitos were captured and Ae. aegypti was much more frequent (92.9%) than Ae. albopictus (6.8%). Ae. aegypti females accounted for 71.8% of captured mosquitoes by MosquitTrap© and were the only species found naturally infected with DENV (infection rate=0.81%). Only one Ae. aegypti male, collected by BG-sentinel, was also tested positive for DENV. The peak of DENV-positive mosquitoes coincided the season of the highest incidence of human cases. The most common serotypes detected in mosquitoes were DENV-3 (24%) and DENV-1 (24%), followed by DENV-4 (20%), DENV-2 (8%) and DENV-1 plus DENV4 (4%), while 95% of laboratory-confirmed human infections in the period were due to DENV-4. These contrasting results suggest silent maintenance of DENV serotypes during the epidemics, reinforcing the importance of entomological and viral surveillance in endemic areas.

Larvicidal activity of Ottonia anisum metabolites against Aedes aegypti: A potential natural alternative source for mosquito vector control in Brazil.

BACKGROUND & OBJECTIVES: Aedes aegypti mosquito is the principal vector of the viruses responsible for urban yellow fever, dengue, dengue haemorrhagic fever, as well as Zika and chikungunya in Brazil. The present study was aimed to investigate the insecticidal potential of the extract and fractions of Ottonia anisum, along with special metabolites isolated from it, as natural alternatives against larvae (L3) of Ae. aegypti, vector of potentially deadly tropical infections in Brazil. METHODS: The plant species O. anisum was collected in March 2015, at Xerιm area, in Rio de Janeiro City, Brazil. Crude extracts and the isolated pure compounds were screened for toxicity against Ae. aegypti larvae (L3). Bioassays were performed on 20 larvae (L3) of Ae. aegypti in triplicate. The samples were dissolved in a mixture of acetone and DMSO at final concentrations of 1-200 µg/ml. The toxicity of the solutions was evaluated towards the growth and development of Ae. aegypti larvae till emergence of adults. RESULTS: The crude hexane extract showed 100% larval mortality 24 h after treatment at a concentration of 200 µg/ml. The bioassays using 1-butyl-3,4-methylenedioxybenzene revealed 100% mortality among L3 larvae, 24 h afterthe treatment at a concentration of 30 µg/ml, the LC recorded was 1.6 µg/ml. At concentration of 10 µg/ml, the L3 larval mortality recorded was 92%. INTERPRETATION & CONCLUSION: The metabolite 1-butyl-3,4-methylenedioxybenzene showed potent toxicity against Ae. aegypti larvae. This arylbutanoid agent could be used as a natural alternative adjuvant pesticide, in new compositions that would be environmentally safer.

Surveillance of dengue vectors using spatio-temporal Bayesian modeling.

BACKGROUND: At present, dengue control focuses on reducing the density of the primary vector for the disease, Aedes aegypti, which is the only vulnerable link in the chain of transmission. The use of new approaches for dengue entomological surveillance is extremely important, since present methods are inefficient. With this in mind, the present study seeks to analyze the spatio-temporal dynamics of A. aegypti infestation with oviposition traps, using efficient computational methods. These methods will allow for the implementation of the proposed model and methodology into surveillance and monitoring systems. METHODS: The study area includes a region in the municipality of Rio de Janeiro, characterized by high population density, precarious domicile construction, and a general lack of infrastructure around it. Two hundred and forty traps were distributed in eight different sentinel areas, in order to continually monitor immature Aedes aegypti and Aedes albopictus mosquitoes. Collections were done weekly between November 2010 and August 2012. The relationship between egg number and climate and environmental variables was considered and evaluated through Bayesian zero-inflated spatio-temporal models. Parametric inference was performed using the Integrated Nested Laplace Approximation (INLA) method. RESULTS: Infestation indexes indicated that ovipositing occurred during the entirety of the study period. The distance between each trap and the nearest boundary of the study area, minimum temperature and accumulated rainfall were all significantly related to the number of eggs present in the traps. Adjusting for the interaction between temperature and rainfall led to a more informative surveillance model, as such thresholds offer empirical information about the favorable climatic conditions for vector reproduction. Data were characterized by moderate time (0.29 - 0.43) and spatial (21.23 - 34.19 m) dependencies. The models also identified spatial patterns consistent with human population density in all sentinel areas. The results suggest the need for weekly surveillance in the study area, using traps allocated between 18 and 24 m, in order to understand the dengue vector dynamics. CONCLUSIONS: Aedes aegypti, due to it short generation time and strong response to climate triggers, tend to show an eruptive dynamics that is difficult to predict and understand through just temporal or spatial models. The proposed methodology allowed for the rapid and efficient implementation of spatio-temporal models that considered zero-inflation and the interaction between climate variables and patterns in oviposition, in such a way that the final model parameters contribute to the identification of priority areas for entomological surveillance.

Deforestation and Malaria on the Amazon Frontier: Larval Clustering of Anopheles darlingi (Diptera: Culicidae) Determines Focal Distribution of Malaria.

We performed bimonthly mosquito larval collections during 1 year, in an agricultural settlement in the Brazilian Amazon, as well as an analysis of malaria incidence in neighboring houses. Water collections located at forest fringes were more commonly positive for Anopheles darlingi larvae and Kulldorff spatial analysis pinpointed significant larval clusters at sites directly beneath forest fringes, which were called larval "hotspots." Remote sensing identified 43 "potential" hotspots. Sampling of these areas revealed an 85.7% positivity rate for A. darlingi larvae. Malaria was correlated with shorter distances to potential hotpots and settlers living within 400 m of potential hotspots had a 2.60 higher risk of malaria. Recently arrived settlers, usually located closer to the tip of the triangularly shaped deforestation imprints of side roads, may be more exposed to malaria due to their proximity to the forest fringe. As deforestation progresses, transmission decreases. However, forest remnants inside deforested areas conferred an increased risk of malaria. We propose a model for explaining frontier malaria in the Amazon: because of adaptation of A. darlingi to the forest fringe ecotone, humans are exposed to an increased transmission risk when in proximity to these areas, especially when small dams are created on naturally running water collections.

Better to Be in Bad Company than to Be Alone? Aedes Vectors Respond Differently to Breeding Site Quality in the Presence of Others.

This study focuses on two competing species, Aedes aegypti and Aedes albopictus (Diptera: Culicidae), both invasive mosquitoes of the New World. Context-specific competition between immature forms inside containers seems to be an important determinant of the coexistence or displacement of each species in different regions of the world. Here, competition experiments developed at low density (one, two or three larvae) and receiving four different resource food concentration, were designed to test whether Ae. albopictus and Ae. aegypti respond differently to competition, and whether competition can be attributed to a simple division of resources. Three phenotypic traits - larval development, adult survival under starvation and wing length - were used as indicators of performance. Larvae of neither species were limited by resource concentration when they were alone, unlike when they developed with competitors. The presence of conspecifics affected Ae. aegypti and Ae. albopictus, inducing slower development, reduced survival and wing length. The response to resource limitation was different when developing with heterospecifics: Ae. aegypti developing with one heterospecific showed faster development, producing smaller adults with shorter lives, while in the presence of two competitors, development increased and adults lived longer. Aedes albopictus demonstrated a better performance when developing with heterospecifics, with no loss in their development period and improved adult survival. Overall, our results suggest that response to competition can not simply be attributed to the division of resources, and that larvae of both species presented large phenotypic plasticity in their response to the presence or absence of heterospecifics and conspecifics.

Surveillance of Aedes aegypti: comparison of house index with four alternative traps.

INTRODUCTION: The mosquito Aedes aegypti, vector of dengue, chikungunya and yellow fever viruses, is an important target of vector control programs in tropical countries. Most mosquito surveillance programs are still based on the traditional household larval surveys, despite the availability of new trapping devices. We report the results of a multicentric entomological survey using four types of traps, besides the larval survey, to compare the entomological indices generated by these different surveillance tools in terms of their sensitivity to detect mosquito density variation. METHODS: The study was conducted in five mid-sized cities, representing variations of tropical climate regimens. Surveillance schemes using traps for adults (BG-Sentinel, Adultrap and MosquiTRAP) or eggs (ovitraps) were applied monthly to three 1 km(2) areas per city. Simultaneously, larval surveys were performed. Trap positivity and density indices in each area were calculated and regressed against meteorological variables to characterize the seasonal pattern of mosquito infestation in all cities, as measured by each of the four traps. RESULTS: The House Index was consistently low in most cities, with median always 0. Traps rarely produced null indices, pointing to their greater sensitivity in detecting the presence of Ae. aegypti in comparison to the larval survey. Trap positivity indices tend to plateau at high mosquito densities. Despite this, both indices, positivity and density, agreed on the seasonality of mosquito abundance in all cities. Mosquito seasonality associated preferentially with temperature than with precipitation even in areas where temperature variation is small. CONCLUSIONS: All investigated traps performed better than the House Index in measuring the seasonal variation in mosquito abundance and should be considered as complements or alternatives to larval surveys. Choice between traps should further consider differences of cost and ease-of-use.

Seasonal and nonseasonal dynamics of Aedes aegypti in Rio de Janeiro, Brazil: fitting mathematical models to trap data.

Mathematical models suggest that seasonal transmission and temporary cross-immunity between serotypes can determine the characteristic multi-year dynamics of dengue fever. Seasonal transmission is attributed to the effect of climate on mosquito abundance and within host virus dynamics. In this study, we validate a set of temperature and density dependent entomological models that are built-in components of most dengue models by fitting them to time series of ovitrap data from three distinct neighborhoods in Rio de Janeiro, Brazil. The results indicate that neighborhoods differ in the strength of the seasonal component and that commonly used models tend to assume more seasonal structure than found in data. Future dengue models should investigate the impact of heterogeneous levels of seasonality on dengue dynamics as it may affect virus maintenance from year to year, as well as the risk of disease outbreaks.

On the use of classic epidemiological formulae for estimating the intensity of endemic malaria transmission by vectors in the Amazon.

Although various reports have described entomological inoculation rates of malaria vector species, most were limited to providing descriptive field data. Here, we report biting rates and survival data for two important malaria vectors in the Amazon, Anopheles darlingi (Root) and Anopheles albitarsis E (Lynch-Arribalzaga) (Diptera: Culicidae), in the state of Roraima, Brazil. We calculated theoretical sporozoite infection rates and critical vector biting rates for these species during 1 year, comprising six bimestrial collections. Anopheles darlingi had higher sporozoite rates and lower critical biting rates, indicating that it would be the more efficient vector at the beginning of epidemic malaria transmission. Our data, together with compiled information from the literature in the Amazon, suggest that epidemic malaria transmission may be initiated by the primary vector, such as A. darlingi, while secondary vectors, such as A. albitarsis E, may only become epidemiologically important when there is an increase in the prevalence of human malaria. We propose that mathematical modeling may be able to quantify the relative importance of secondary vector species in malaria epidemiology.

Mosquito Anthropophily: implications on malaria transmission in the northern Brazilian Amazon

Seasonal variation of adults and larvae of anophelines was studied during 2003 and 2004 in Roraima State, located in the Northern region of Brazilian Amazon. Species diversity increased with distance of capture to human dwellings. Greater diversity was found in extradomiciliary collections than in peridomiciliary or intradomiciliary. A significant association between Anopheles darlingi Root and An. albitarsis (s.l.) Arribálzaga (Diptera: Culicidae) breeding sites and the proximity to human dwellings was observed. Malaria Sporozoite Antigen Panel Assay (VectestTM Malaria) indicated An. albitarsis s.l. as one of the local vectors in the studied area. In this study, an index to describe the anthropophilic behavior of each anopheline species is proposed.

Mosquito anthropophily: implications on malaria transmission in the Northern Brazilian Amazon.

Seasonal variation of adults and larvae of anophelines was studied during 2003 and 2004 in Roraima State, located in the Northern region of Brazilian Amazon. Species diversity increased with distance of capture to human dwellings. Greater diversity was found in extradomiciliary collections than in peridomiciliary or intradomiciliary. A significant association between Anopheles darlingi Root and An. albitarsis (s.l.) Arribálzaga (Diptera: Culicidae) breeding sites and the proximity to human dwellings was observed. Malaria Sporozoite Antigen Panel Assay (VectestTM Malaria) indicated An. albitarsis s.l. as one of the local vectors in the studied area. In this study, an index to describe the anthropophilic behavior of each anopheline species is proposed.

Occurrence of Toxorhynchites guadeloupensis (Dyar Knab) in oviposition trap of Aedes aegypti (L.) (Diptera: Culicidae).

Toxorhynchites guadeloupensis (Dyar Knab), a poorly known mosquito species, was observed preying upon Aedes aegypti (L.) larvae, in an oviposition trap placed for routine dengue entomological surveillance, during 2003-2004 in the urban area of Boa Vista, Roraima, Brazil. This is the first report for Tx. guadeloupensis using Ae. aegypti oviposition traps as breeding places. This finding may have important consequences in the epidemiology and local dengue control since Ae. aegypti density is a basic variable in dengue prediction. Whether predation of Ae aegypti by Tx. guadeloupensis in the Amazon is of significance, is a question to be examined. Also, larval predation may be a cause for underestimation of the actual Ae aegypti numbers. Together these hypotheses need to be better investigated as they are directly related to dengue epidemiology, to the success of any outbreak prediction and surveillance program.

Occurrence of Toxorhynchites guadeloupensis (Dyar & Knab) in oviposition trap of Aedes aegypti (L) (Diptera: Culicidae)

Toxorhynchites guadeloupensis (Dyar & Knab), a poorly known mosquito species, was observed preying upon Aedes aegypti (L.) larvae, in an oviposition trap placed for routine dengue entomological surveillance, during 2003-2004 in the urban area of Boa Vista, Roraima, Brazil. This is the first report for Tx. guadeloupensis using Ae. aegypti oviposition traps as breeding places. This finding may have important consequences in the epidemiology and local dengue control since Ae. aegypti density is a basic variable in dengue prediction. Whether predation of Ae aegypti by Tx. guadeloupensis in the Amazon is of significance, is a question to be examined. Also, larval predation may be a cause for underestimation of the actual Ae aegypti numbers. Together these hypotheses need to be better investigated as they are directly related to dengue epidemiology, to the success of any outbreak prediction and surveillance program.

Larvas de Toxorhynchites guadeloupensis (Dyar & Knab), espécie raramente encontrada, foram observadas predando larvas de Aedes aegypti (L.), em uma armadilha para ovos de monitoração rotineira para o controle do dengue na área urbana da cidade de Boa Vista, RO, em 2003-2004. Essa observação constitui o primeiro registro de Tx. guadeloupensis em armadilhas para ovos de Ae. aegypti e pode ocasionar repercussões importantes na epidemiologia e no controle local do dengue, uma vez que a densidade de Ae. aegypti em um dado momento constitui uma das variáveis básicas na predição da doença. Se a predação de Ae. aegypti por Tx. guadeloupensis na Amazônia é significativa, é uma questão que precisa ser melhor investigada. A predação larval pode levar ainda a uma subestimativa do número real de espécimens de Ae aegypti preditas pela amostragem. Juntas, essas hipóteses precisam de atenção visto que estão diretamente relacionadas à epidemiologia do dengue, ao sucesso de qualquer predição e programas de controle.

Distribution of dengue vectors in neighborhoods with different urbanization types of Manaus, state of Amazonas, Brazil.

Aedes aegypti and Ae. albopictus are vectors of dengue viruses, which cause endemic disease in the city of Manaus, capital of the state of Amazonas, Brazil. More than 53 thousand cases have been registered in this city since the first epidemic in 1998. We evaluated the hypothesis that different ecological conditions result in different patterns of vector infestation in Manaus, by measuring the infestation level in four neighborhoods with different urbanization patterns, during the rainy (April), dry (August), and transitional (November) seasons. Ae. aegypti predominated throughout the study areas and sampling periods, representing 86% of all specimens collected in oviposition traps. High frequencies of houses positive for both species were observed in all studied sites, with Ae. aegypti present in more than 84% of the houses in all seasons. Ae. albopictus, on the other hand, showed more spatial and temporal variation in abundance. We found no association between infestation level and house traits. This study highlights the homogeneity of dengue vector distribution in Manaus.

Distribution of dengue vectors in neighborhoods with different urbanization types of Manaus, state of Amazonas, Brazil

Aedes aegypti and Ae. albopictus are vectors of dengue viruses, which cause endemic disease in the city of Manaus, capital of the state of Amazonas, Brazil. More than 53 thousand cases have been registered in this city since the first epidemic in 1998. We evaluated the hypothesis that different ecological conditions result in different patterns of vector infestation in Manaus, by measuring the infestation level in four neighborhoods with different urbanization patterns, during the rainy (April), dry (August), and transitional (November) seasons. Ae. aegypti predominated throughout the study areas and sampling periods, representing 86 percent of all specimens collected in oviposition traps. High frequencies of houses positive for both species were observed in all studied sites, with Ae. aegypti present in more than 84 percent of the houses in all seasons. Ae. albopictus, on the other hand, showed more spatial and temporal variation in abundance. We found no association between infestation level and house traits. This study highlights the homogeneity of dengue vector distribution in Manaus.

Tetrahymenidae infection in mosquito populations in a malaria-endemic region of the Amazon.

In a field survey performed in a malaria-endemic region of Northern Amazon, Brazil, we encountered ciliate protozoa of the family Tetrahymenidae infecting adults and larvae of the following mosquito species: Culex sp., Anopheles albitarsis l.s., Anopheles strodei, Anopheles mattogrossensis, Anopheles darlingi, and Anopheles oswaldoi l.s. Based on morphological features and life style, we have tentatively identified the parasite as Lambornella sp. The association appears pathogenic for the mosquito. Prevalence of infection in both larvae and adults was higher in the dry season.

Convergent habitat segregation of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in southeastern Brazil and Florida.

During the rainy season of 2001, the incidence of the dengue vectors Aedes aegypti and Ae. albopictus was examined in different habitats of two cities (Rio de Janeiro and Nova Iguaçu) in Rio de Janeiro State, Brazil, and in two cities (Palm Beach and Boca Raton) in Florida. Oviposition trap collections were performed in urban, suburban, and rural habitats in both areas. Our hypothesis that the abundances and frequencies of occurrence of Ae. aegypti and Ae albopictus are affected in opposite ways by increasing urbanization was only partially supported. City, habitat, and their interaction significantly affected the abundance of both species. Cities with high abundance of Ae. aegypti also had a high abundance of Ae. albopictus. The two species were most abundant in the cities of Rio de Janeiro state and the lowest in Boca Raton. Habitat had a significant but opposite effect on the abundances of Ae. aegypti and Ae. albopictus. In general, Ae. aegypti was most prevalent in highly urbanized areas and Ae. albopictus in rural, suburban, and vegetated urban areas in Rio de Janeiro state and Florida. However, abundances of the two species were similar in most suburban areas. Analyses of frequencies of occurrence showed an unexpected high level of co-occurrence of both species in the same oviposition trap. Despite the different geographical origins of Ae. albopictus in Brazil and the United States, the habitats used by this recent invader are remarkably similar in the two countries.

Dengue virus type 3 isolation from Aedes aegypti in the municipality of Nova Iguaçu, State of Rio de Janeiro.

In a prospective field study conducted from July 2000 to June 2001, adult Aedes aegypti and Ae. albopictus mosquitoes were caught from the municipality of Nova Iguaçu, State of Rio de Janeiro, Brazil. Virus isolation in Ae. albopictus clone C6/36 cell line and a semi-nested reverse transcription-polymerase chain reaction detected only dengue virus type 3 in three pools of Ae. aegypti, despite the co-circulation of DEN-1, DEN-2 and DEN-3 serotypes in that area. No viruses were detected in Ae. albopictus mosquitoes. This virological surveillance consists in a sentinel system alerting for dengue outbreaks.