Effects of environment, dietary regime and ageing on the dengue vector microbiota: evidence of a core microbiota throughout Aedes aegypti lifespan

Abstract Mosquito midgut microbiota is a key component of vector competence, as gut bacteria can disturb pathogen development. In this study, we addressed the microbiota composition of Aedes aegypti during its lifespan, under field conditions. We also investigated the possible effects of environment, dietary regime and ageing on the gut community composition. We employed culture independent and dependent approaches to characterise vector microbiota. There was evidence of a lifelong stable core microbiota after mosquitoes were released into an urban settlement, where they presumably fed on a range of vertebrate hosts and carbohydrate sources. This core was formed mainly of bacteria belonging to the genera Pseudomonas, Acinetobacter, Aeromonas and Stenotrophomonas and to the families Oxalobacteraceae, Enterobacteriaceae and Comamonadaceae. We showed that both dietary regime and age were associated with the abundance of some bacterial groups in the Ae. aegypti microbiota. The majority of the bacterial groups we identified have been detected in the midgut of Ae. aegypti from laboratory and wild populations, indicating a possible core microbiota associated with this mosquito species. Our findings suggest that Ae. aegypti harbours a stable bacterial community during its adult life, similar to mosquito populations from distinct geographic areas, which may be further explored for arbovirus biocontrol strategies.

She’s a femme fatale: low-density larval development produces good disease vectors

Two hypotheses for how conditions for larval mosquitoes affect vectorial capacity make opposite predictions about the relationship of adult size and frequency of infection with vector-borne pathogens. Competition among larvae produces small adult females. The competition-susceptibility hypothesis postulates that small females are more susceptible to infection and predicts frequency of infection should decrease with size. The competition-longevity hypothesis postulates that small females have lower longevity and lower probability of becoming competent to transmit the pathogen and thus predicts frequency of infection should increase with size. We tested these hypotheses for Aedes aegypti in Rio de Janeiro, Brazil, during a dengue outbreak. In the laboratory, longevity increases with size, then decreases at the largest sizes. For field-collected females, generalised linear mixed model comparisons showed that a model with a linear increase of frequency of dengue with size produced the best Akaike’s information criterion with a correction for small sample sizes (AICc). Consensus prediction of three competing models indicated that frequency of infection increases monotonically with female size, consistent with the competition-longevity hypothesis. Site frequency of infection was not significantly related to site mean size of females. Thus, our data indicate that uncrowded, low competition conditions for larvae produce the females that are most likely to be important vectors of dengue. More generally, ecological conditions, particularly crowding and intraspecific competition among larvae, are likely to affect vector-borne pathogen transmission in nature, in this case via effects on longevity of resulting adults. Heterogeneity among individual vectors in likelihood of infection is a generally important outcome of ecological conditions impacting vectors as larvae.

Discrepancies between Aedes aegypti identification in the field and in the laboratory after collection with a sticky trap

Currently, sticky traps are regularly employed to assist in the surveillance of Aedes aegypti infestation. We tested two alternative procedures for specimen identification performed by local health agents: directly in the field, as recommended by certain manufacturers, or after transportation to the laboratory. A total of 384 sticky traps (MosquiTRAP) were monitored monthly during one year in four geographically representative Brazilian municipalities. When the same samples were inspected in the field and in the laboratory, large differences were noted in the total number of mosquitoes recorded and in the number of specimens identified as Ae. aegypti by both procedures. Although field identification has the potential to speed vector surveillance, these results point to uncertainties in the evaluated protocol.

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

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.

Evaluation of RbCl and CrCl3 as markers of Triatoma brasiliensis (Hemiptera: Reduviidae) nymphs: persistence and influence of Rb and Cr on triatomine biology

In order to mark Triatoma brasiliensis, the vector of Chagas disease in Brazil, two chemical compounds, rubidium chloride (RbCl) and chromium chloride (CrCl3), were tested. First, 199 N2-N5 nymphs were fed on blood with 0.025M RbCl. Rb marker positivity ranged from 2.5 percent (N3)-70 percent (N2), with a maximum persistence of 98 days. Second, 265 N2-N5 nymphs were fed on blood containing 0.0015M CrCl3. Cr marker positivity ranged up to 93 percent (N5), with a maximum persistence of 119 days. Finally, we blood fed 213 T. brasiliensis to investigate whether CrCl3 altered the biology of this insect. The developmental time of T. brasiliensis was unaltered, but the survival of the Cr-marked group was lower than that of the control group. Differences in the mean fecundity of the control (mean of 156.1) and experimental (mean of 135.6) groups were not statistically significant and 100 percent of the egg batches of females Cr-marked as nymphs were positive. In conclusion, CrCl3 is a useful tool for marking T. brasiliensis nymphs due to its high positivity and persistence.

Presumed unconstrained dispersal of Aedes aegypti in the city of Rio de Janeiro, Brazil / Dispersão de Aedes aegypti em local presumidamente sem barreira limitante ao vôo na cidade do Rio de Janeiro / Dispersión ilimitada de Aedes aegypti en la ciudad de Río de Janeiro, Brasil

OBJECTIVE: To evaluate dispersal of Aedes aegypti females in an area with no container manipulation and no geographic barriers to constrain mosquito flight. METHODS: A mark-release-recapture experiment was conducted in December 2006, in the dengue endemic urban district of Olaria in Rio de Janeiro, Southeastern Brazil, where there is no evident obstacle to the dispersal of Ae. aegypti females. Mosquito traps were installed in 192 houses (96 Adultraps and 96 MosquiTRAPs). RESULTS: A total of 725 dust-marked gravid females were released and recapture rate was 6.3 percent. Ae. aegypti females traveled a mean distance of 288.12 m and their maximum displacement was 690 m; 50 percent and 90 percent of females flew up to 350 m and 500.2 m, respectively. CONCLUSIONS: Dispersal of Ae. aegypti females in Olaria was higher than in areas with physical and geographical barriers. There was no evidence of a preferred direction during mosquito flight, which was considered random or uniform from the release point.

OBJETIVO: Avaliar a dispersão de fêmeas de Aedes aegypti em uma área onde não houve nenhuma interferência quanto à redução de potenciais criadouros e não há barreira geográfica que limite o vôo dos mosquitos. MÉTODOS: Um experimento de marcação-soltura-recaptura foi realizado em dezembro de 2006, no bairro urbano Olaria, endêmico para dengue na cidade do Rio de Janeiro (RJ), onde não há obstáculos evidentes à dispersão de fêmeas de Ae. aegypti. Armadilhas para captura de mosquitos foram instaladas em 192 residências (96 Adultraps e 96 MosquiTraps). RESULTADOS: Foram soltas 725 fêmeas grávidas marcadas com pó fluorescente e a taxa de recaptura foi de 6,3 por cento. Fêmeas de Ae. aegypti dispersaram em média 288,12 m do ponto de soltura e o deslocamento máximo foi de 690 m; 50 por cento e 90 por cento das fêmeas voaram até 350 m e 500,2 m respectivamente. CONCLUSÕES: A dispersão de fêmeas de Ae. aegypti em Olaria foi maior que em áreas com barreiras físicas e/ou geográficas. Não houve evidências de preferência de direção do vôo dos mosquitos, o qual foi considerado randômico ou uniforme a partir do ponto de soltura.

OBJETIVO: Evaluar la dispersión de hembras de Aedes aegypti en un área sin manipulación de recipientes y sin barreras geográficas que limiten el vuelo del mosquito. MÉTODOS: Se realizó un experimento de marcaje y recaptura en diciembre de 2006, en el distrito urbano Olaria, Rio de Janeiro (Sureste de Brasil), área endémica para dengue, donde no hay obstáculo evidente para la dispersión de hembras de Ae. aegypti. Trampas para mosquitos fueron instaladas en 192 casas (96 Adultraps y 96 MosquiTraps). RESULTADOS: Un total de 725 hembras grávidas marcadas con polvo fueron liberadas y recapturadas siendo la tasa de 6,3 por ciento. Las hembras de Ae. Aegypti viajaron una distancia promedio de 288,12 m y su máximo desplazamiento fue de 690 m; 50 por ciento y 90 por ciento de las hembras volaron hasta los 350 m y 500,2 m, respectivamente. CONCLUSIONES: Dispersión de las hembras de Ae. aegypti en Olaria fue mayor que en las áreas con barreras físicas y geográficas. No hube evidencia de una dirección preferida durante el vuelo del mosquito, por lo que se considera aleatorio y uniforme desde el punto de liberación.

Calculating the survival rate and estimated population density of gravid Aedes aegypti (Diptera, Culicidae) in Rio de Janeiro, Brazil / Determinação da taxa de sobrevivência e estimativa da densidade populacional de fêmeas grávidas de Aedes aegypti (Diptera, Culicidae) no Rio de Janeiro, Brasil

Population size and daily survival rates of disease vectors are important determinants of vectorial capacity. A mark-release-recapture experiment was conducted in a dengue endemic urban neighborhood of Rio de Janeiro, Brazil, to estimate population size, survival rate and vectorial capacity of Aedes aegypti females using back-pack aspirators and gravid sticky traps (MosquiTRAP). Estimations of the gravid female population size were different when using data gathered from just the MosquiTRAP (3,505 individuals) or aspirator (1,470). However Ae. aegypti survival rates and longevity were similar irrespective of the method of capture. Up to 26.3 percent of released females would be able to survive for more than 10 days, the length of time of the extrinsic incubation period. Vectorial capacity value ranged between 0.01567 and 0.4215 and the basic reproductive number (R0) was estimated to be between 0.0695 and 1.88.

O tamanho populacional e a taxa de sobrevivência de vetores de doenças são importantes componentes da capacidade vetorial. Um experimento de marcação-soltura-recaptura foi conduzido numa área urbana endêmica de dengue no Rio de Janeiro, Brasil, para estimar o tamanho populacional, taxa de sobrevivência diária e capacidade vetorial de fêmeas de Aedes aegypti; para tal, usaram-se aspirador costal e armadilha para fêmeas grávidas de Ae. aegypti (MosquiTRAP). Estimativas do tamanho populacional de fêmeas grávidas foram diferentes quando foram analisados dados coletados apenas na MosquiTRAP (3.505 indivíduos) e aspiradores (1.470). Por outro lado, taxas de sobrevivência e longevidade de Ae. aegypti foram semelhantes independentemente do método de captura empregado. Até 26,3 por cento das fêmeas seriam capazes de sobreviver mais do que dez dias, tempo equivalente à duração do período de incubação extrínseco. O cálculo da capacidade vetorial revelou valores entre 0,01567 e 0,4215, e o número básico de reprodução foi estimado entre 0,0695 e 1,88.

Mosquito traps designed to capture Aedes aegypti (Diptera: Culicidae) females: preliminary comparison of Adultrap, MosquiTRAP and backpack aspirator efficiency in a dengue-endemic area of Brazil

In this report, the efficiency of Adultrap under field conditions is compared to a CDC backpack aspirator and to MosquiTRAP. An urban dengue-endemic area of Rio de Janeiro was selected to evaluate the efficiency of mosquito traps in capturing Aedes aegypti females. Adultrap and aspirator captured similar numbers of Ae. aegypti females, with the former showing high specificity to gravid individuals (93.6 percent). A subsequent mark-release-recapture experiment was conducted to evaluate Adultrap and MosquiTRAP efficiency concomitantly. With a 6.34 percent recapture rate, MosquiTRAP captured a higher mean number of female Ae. aegypti per trap than Adultrap (Ç2 = 14.26; df = 1; p < 0,05). However, some MosquiTRAPs (28.12 percent) contained immature Ae. aegypti after 18 days of exposure in the field and could be pointed as an oviposition site for female mosquitoes. Both trapping methods, designed to collect gravid Ae. aegypti females, seem to be efficient, reliable and may aid routine Ae. aegypti surveillance.

Variation in Aedes aegypti(Diptera: Culicidae) container productivity in a slum and a suburban district of Rio de Janeiro during dry and wet seasons

Seasonal variation in container productivity and infestation levels by Aedes aegypti were evaluated in two areas with distinct levels of urbanization degrees in Rio de Janeiro, a slum and a suburban neighborhood. The four most productive containers can generate up to 90 percent of total pupae. Large and open-mouthed containers, such as water tanks and metal drums, located outdoors were the most productive in both areas, with up to 47.49 percent of total Ae. aegypti pupae collected in the shaded sites in the suburban area. Water-tanks were identified as key containers in both areas during both the dry and rainy seasons. Container productivity varied according to seasons and urbanization degree. However, the mean number of pupae per house was higher in the suburban area, but not varied between seasons within each area (P > 0.05). High infestation indexes were observed for both localities, with a house index of 20.5-21.14 in the suburban and of 9.56-11.22 in the urban area. This report gives potential support to a more focused and cost-effective Ae. aegypti control in Rio de Janeiro.

Field evaluation of effectiveness of the BG-Sentinel, a new trap for capturing adult Aedes aegypti (Diptera: Culicidae)

In recent years, the development of new tools to gather field information about vector ecological parameters has increased. This report evaluated the BG-Sentinel Trap (BGS-Trap), a promising new attempt to improve collection of the dengue vector, Aedes aegypti. The efficacy of the BGS-Trap was compared with the CDC backpack aspirator, one of the commonest used methods for capturing adult mosquitoes. BGS-Traps captured significantly more Ae. aegypti males (chi2 = 21.774, df = 1, P < 0.05) and females (chi2 = 56.007, df = 1, P < 0.05) than CDC aspirator during all days of field collection. However, CDC aspirator was significantly more efficient to capture Culex quinquefasciatus males (chi2 = 5.681, df = 1, P < 0.05) and females (chi2 = 6.553, df = 1, P < 0.05). BGS-Traps captured host-seeking females (varying between 68.75 to 89.8 percent) in detriment of females in other behavioral and physiological stages. BGS-Traps proved to be efficient and can be used for monitoring adult mosquito populations.

Efficiency of rubidium marking in Aedes albopictus (Diptera: Culicidae): preliminary evaluation on persistence of egg labeling, survival, and fecundity of marked female

Rubidium chloride (RbCl) has been used for the study of vector biology and behavior, although the efficacy of marking, egg production, and survivorship of marked females have been poorly studied. Four concentrations of RbCl were tested, among which 0.025 M was the best for marking Aedes albopictus: more than 80 percent of egg batches of females fed once with blood containing RbCl were marked; Rb-marked egg batches, interspersed with non marked ones were recovered until 61 days after a blood meal containing RbCl followed by non marked meals; RbCl was essentially detected in the abdomen of marked females, whose egg production and survivorship did not differ from non marked ones, at least in the three weeks following the Rb-marked blood meal.