Mariana Rocha David

Container productivity, daily survival rates and dispersal of Aedes aegypti mosquitoes in a high income dengue epidemic neighbourhood of Rio de Janeiro: presumed influence of differential urban structure on mosquito biology

Different urban structures might affect the life history parameters of Aedes aegypti and, consequently, dengue transmission. Container productivity, probability of daily survival (PDS) and dispersal rates were estimated for mosquito populations in a high income neighbourhood of Rio de Janeiro. Results were contrasted with those previously found in a suburban district, as well as those recorded in a slum. After inspecting 1,041 premises, domestic drains and discarded plastic pots were identified as the most productive containers, collectively holding up to 80% of the total pupae. In addition, three cohorts of dust-marked Ae. aegypti females were released and recaptured daily using BGS-Traps, sticky ovitraps and backpack aspirators in 50 randomly selected houses; recapture rate ranged from 5-12.2% within cohorts. PDS was determined by two models and ranged from 0.607-0.704 (exponential model) and 0.659-0.721 (non-linear model), respectively. Mean distance travelled varied from 57-122 m, with a maximum dispersal of 263 m. Overall, lower infestation indexes and adult female survival were observed in the high income neighbourhood, suggesting a lower dengue transmission risk in comparison to the suburban area and the slum. Since results show that urban structure can influence mosquito biology, specific control strategies might be used in order to achieve cost-effective Ae. aegypti control.

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.

Bionomics of Culex quinquefasciatus within urban areas of Rio de Janeiro, Southeastern Brazil

OBJECTIVE To evaluate density, parity rates, daily survival and longevity of natural populations of Culex quinquefasciatus in three neighborhoods with distinct socio-economic and infrastructure profiles. METHODS Mosquito collections of the Culex quinquefasciatus species were performed weekly during two four month periods, from August to November 2008 (spring) and March to June 2009 (fall), in a favela (slum), a suburban area and a middle class area of Rio de Janeiro, Southeastern Brazil. Collections were performed with backpack aspirators, in 20 randomly selected houses in each area per week, during 15-20 minutes per house. Ovaries were removed from captured females and classified as initial, intermediary or final stage. Furthermore, females were dissected for determination of parity based on the condition of the tracheal system. Mosquito survival rate and longevity were estimated on a per month basis for each neighborhood. RESULTS We collected a total of 2,062 Culex quinquefasciatus, but monthly vector density was not correlated with temperature and rainfall. We dissected the ovaries of 625 Culex quinquefasciatus, and overall, there was a higher proportion of nulliparous females during the dryer months, while gravid females were more frequent in rainy months. In the middle class neighborhood, the parity rate reached up to 93.75% with survivorship of 0.979. Lower parity and survival rates were obtained in the suburban area (as low as 36.4% parity and 0.711 daily survival). Up to 84.7% of Culex quinquefasciatus females could survive the eight day period needed to complete West Nile Virus incubation. CONCLUSIONS The survival rate of Culex quinquefasciatus varied significantly between the neighborhoods. This suggests that vectorial capacity and disease transmission risk may vary greatly between different urban areas, which is potentially useful information for vector control programs.OBJETIVO: Avaliar densidade, taxa de paridade, sobrevivencia diaria e longevidade de populacoes naturais de Culex quinquefasciatus de tres localidades com diferentes perfis socioeconomicos e de infraestrutura. METODOS: Mosquitos da especie Culex quinquefasciatus foram coletados semanalmente em dois periodos de quatro meses, de agosto a novembro de 2008 e de marco a junho de 2009, em uma comunidade, uma area de suburbio e um bairro de classe media no Rio de Janeiro, RJ. As coletas foram realizadas com o auxilio de aspiradores costais em 20 domicilios, aleatoriamente selecionados por semana e por area, durante 15-20 minutos por domicilio. As femeas capturadas tiveram os ovarios removidos e classificados em estagios inicial, intermediario e final. Adicionalmente, as femeas foram dissecadas para determinacao da paridade de acordo com as condicoes do sistema traqueal. A taxa de sobrevivencia e a longevidade das femeas foram estimadas por mes, para cada localidade. RESULTADOS: Foram coletados 2.062 Culex quinquefasciatus, porem a densidade mensal deste vetor nao apresentou correlacao com temperatura e precipitacao. Dissecamos os ovarios de 625 Culex quinquefasciatus e obtivemos maior proporcao de femeas nuliparas durante os meses mais secos, enquanto femeas gravidas foram mais frequentes nos meses chuvosos. A taxa de paridade foi de ate 93,75% no bairro de classe media, com sobrevivencia de 0,979. Menores valores de paridade e sobrevivencia foram obtidos no suburbio (comecando em 36,4% de paridade e 0,711 de sobrevivencia diaria). Ate 84,7% das femeas de Culex quinquefasciatus poderiam sobreviver ao periodo de oito dias, necessario para completar o periodo de incubacao do virus da febre do Nilo Ocidental. CONCLUSOES: A taxa de sobrevivencia de Culex quinquefasciatus variou significativamente entre os bairros, o que sugere que a capacidade vetorial e o risco de transmissao de doencas podem variar entre diferentes areas urbanas, informacao relevante para o planejamento dos programas de controle de vetores.

The impact of insecticide applications on the dynamics of resistance: The case of four Aedes aegypti populations from different Brazilian regions

Background In the tropics, the utilization of insecticides is still an important strategy for controlling Aedes aegypti, the principle vector of dengue, chikungunya and Zika viruses. However, increasing insecticide resistance in Ae. aegypti populations might hinder insecticide efficacy on a long-term basis. It will be important to understand the dynamics and evolution of insecticide resistance by assessing its frequency and the mechanisms by which it occurs. Methodology/Principal findings The insecticide resistance status of four Brazilian Ae. aegypti populations was monitored. Quantitative bioassays with the major insecticides employed in the country was performed: the adulticide deltamethrin (a pyrethroid—PY) and the larvicides, temephos (an organophosphate) and diflubenzuron (a chitin synthesis inhibitor). Temephos resistance was detected in all populations although exhibiting a slight decrease over time probably due to the interruption of field use. All vector populations were susceptible to diflubenzuron, recently introduced in the country to control Ae. aegypti. Resistance against deltamethrin was extremely high in three populations. Molecular assays investigated substitutions in the voltage gated sodium channel (NaV), the PY target site, at positions 1011, 1016 and 1534. Elevated frequencies of substitutions Val1016Ile and Phe1534Cys related to high PY resistance levels were identified. Biochemical assays detected alterations in the activities of two detoxifying enzyme classes related to metabolic resistance, glutathion-S-transferases and esterases. The results obtained were evaluated in the context of both recent insecticide use and the records of dengue incidence in each locality. Conclusions/Significance The four Ae. aegypti populations evaluated were resistant to the neurotoxic insecticides, temephos and deltamethrin. However, they were still susceptible to diflubenzuron. A probable correlation between adult insect resistance to PY and the domestic application of insecticides is discussed, pointing to the need for awareness measures regarding the correct utilization by citizens. This work aims to contribute to the efficient and rational management of Ae. aegypti control of both larvae and adults.

Rapid, noninvasive detection of Zika virus in Aedes aegypti mosquitoes by near-infrared spectroscopy

We report the first use of near-infrared spectroscopy for rapid, low-cost, noninvasive detection of Zika virus in Aedes aegypti. The accelerating global spread of arboviruses, such as Zika virus (ZIKV), highlights the need for more proactive mosquito surveillance. However, a major challenge during arbovirus outbreaks has been the lack of rapid and affordable tests for pathogen detection in mosquitoes. We show for the first time that near-infrared spectroscopy (NIRS) is a rapid, reagent-free, and cost-effective tool that can be used to noninvasively detect ZIKV in heads and thoraces of intact Aedes aegypti mosquitoes with prediction accuracies of 94.2 to 99.3% relative to quantitative reverse transcription polymerase chain reaction (RT-qPCR). NIRS involves simply shining a beam of light on a mosquito to collect a diagnostic spectrum. We estimated in this study that NIRS is 18 times faster and 110 times cheaper than RT-qPCR. We anticipate that NIRS will be expanded upon for identifying potential arbovirus hotspots to guide the spatial prioritization of vector control.

The impact of the age of first blood meal and Zika virus infection on Aedes aegypti egg production and longevity

The impact of senescence and pathogen infection on Aedes aegypti life-history traits remains poorly understood. This laboratory study focused on the impact of Zika virus (ZIKV) infection and the age of first blood intake on blood meal and clutch sizes, and more importantly on the egg production ratio per μL of blood. Three groups of ZIKV-infected and uninfected Ae. aegypti females that received their first blood meal at 7 (young feeders), 14 (mature feeders) and 21 days old (old feeders) were monitored daily for survival and received a blood meal free of ZIKV once a week. The number of eggs laid per female were registered 3–4 days after blood feeding. Infection by ZIKV and age of feeding produced a strong negative impact on survival and oviposition success (e.g. likelihood of laying at least one egg per gonotrophic cycle). Interestingly, clutch size presented a dramatic reduction on uninfected mosquitoes, but raised from 36.5 in clutch1 to 55.1 eggs in clutch 3. Blood meal size remained stable in uninfected females, while a slight increase was observed for the infected counterparts. In uninfected Ae. aegypti, egg production was strongly affected by the age of feeding with younger females laying three times more eggs than when older. On the other hand, ZIKV-infected mosquitoes had a constant but low egg production. Overall, mosquito senescence and ZIKV infection had an impact on mosquito egg production by causing a sharp decrease in the number of eggs along the clutches for uninfected mosquitoes and a slight increase for infected mosquitoes. Despite some study limitations, our results contribute to a better understanding of the effects of mosquito aging and pathogen infection on the vectorial capacity of Ae. aegypti.

Distribution and breeding sites of Aedes aegypti and Aedes albopictus in 32 urban/peri-urban districts of Mozambique: implication for assessing the risk of arbovirus outbreaks

Background Aedes-borne arboviruses have emerged as an important public health problem worldwide and, in Mozambique, the number of cases and its geographical spread have been growing. However, information on the occurrence, distribution and ecology of Aedes aegypti and Ae. albopictus mosquitoes remain poorly known in the country. Methods Between March and April 2016, a cross-sectional study was conducted in 32 districts in Mozambique to determine the distribution and breeding sites of Ae. aegypti and Ae. albopictus. Larvae and pupae were collected from a total of 2,807 water-holding containers using pipette, dipper, funnel and sweeping procedures, depending on the container type and location. Both outdoor and indoor water-holding containers were inspected. The immature forms were reared to adults and the identifications of the mosquito species was carried out with a stereomicroscope using a taxonomic key. Results Aedes aegypti was found in every district sampled, while Ae. albopictus was only found in Moatize district, situated in Tete Province in the central part of the country. Six hundred and twenty-eight of 2,807 (22.4%) containers were positive for Ae. aegypti but only one (0.03%) was positive for Ae. albopictus. The Container Index (CI) of Aedes was highest in densely populated suburban areas of the central region (260/604; 43.0%), followed by suburban areas in northern areas (228/617; 36.9%) whilst the lowest proportion was found in urbanized southern areas (140/1586; 8.8%). The highest CI of Aedes was found in used tires (448/1268; 35.3%), cement tanks (20/62; 32.3%) and drums (21/95; 22.1%). Conclusion Data from our study showed that Ae. aegypti is present nation-wide, since it occurred in every sampled district, whilst Ae. albopictus had a limited distribution. Therefore, the risk of transmission of dengue and chikungunya is likely to have been underestimated in Mozambique. This study highlights the need for the establishment of a national entomological surveillance program for Aedes spp. in Mozambique in order to gain a better understanding about vector bionomics and to support the development of informed effective vector control strategies.

Insecticide Resistance and Fitness: The Case of Four Aedes aegypti Populations from Different Brazilian Regions

Background Chemical control is still a major strategy to constrain vector density and mitigate pathogen transmission. However, insecticide overuse poses a high selective pressure, favouring the spread of resistance alleles in natural populations. In an insecticide-free environment, a fitness cost is expected in resistant insects when compared to susceptible counterparts. This study investigates whether insecticide resistance to an organophosphate (temephos) and a pyrethroid (deltamethrin) is associated with fitness traits in four Aedes aegypti wild populations sampled every three months over one year. Findings We measured development time from larvae to adult, female survival, wing length, fecundity, and adult resistance to starvation in field insecticide resistant Ae. aegypti populations four times over a year. These results were confronted with resistance levels to temephos and deltamethrin and with potentially related mechanisms, including a kdr mutation in the pyrethroid target site. No differences in fitness cost were found after contrasting mosquitoes from the same population collected throughout a year, irrespective of differences in insecticide resistance levels. Additionally, significant differences were not observed among field populations. However, compared to the reference strain Rockefeller, field females survived significantly less. Moreover, larval development was equal or slower in three out of four field populations. In no case differences were evidenced in starvation tolerance, wing length, and fecundity. Conclusions Overall, field resistant mosquitoes seemed to have a slight fitness disadvantage when compared with the Rockefeller susceptible strain which might represent a potential fitness cost of insecticide resistance. However, after comparing Ae. aegypti from the same population but sampled at different moments, or from different field populations, mosquito life-history traits varied independently of resistance ratios. The metabolic deviations necessary to overcome the adverse effects of insecticides may cause an energy trade-off that affects energy allocation and ultimately basic demands of insect biology. The extent of fitness cost due to insecticide resistance is critical information to delay the evolution of resistance in wild vector populations.

Marking Triatoma brasiliensis, Triatoma pseudomaculata and Rhodnius nasutus Nymphs with Trace Elements: Element Persistence and Effects of Marking on Insect Mortality

Background Field ecologists often rely on mark-release-recapture (MRR) experiments to estimate population dynamics parameters for a given species. In the case of a medically important taxon, i.e., a disease vector, inferences on species survival and dispersal rates are particularly important as they have the potential to provide insights into disease transmission dynamics in endemic areas. Medical entomologists have traditionally used fluorescent dusts to externally mark the cuticle of insects. However, dust marking is usually restricted to the adult life stage because immature insects lose the mark when they molt. Methodology/Principal Findings We evaluated the efficacy of 13 trace elements in marking nymphs of three native Brazilian Chagas disease vectors: Triatoma brasiliensis, Triatoma pseudomaculata, and Rhodnius nasutus. Cr and Cu were detected in over 97% of T. brasiliensis (34/35 31/31 for Cr and Cu), while Cu and Mn were detected in more than 95% of T. pseudomaculata (29/29 for Cu and 28/29 for Mn) tested 120 days after marking. Only Mn marked over 90% of R. nasutus nymphs (38/41). Overall, trace elements had no negative effects on T. pseudomaculata longevity, but As-marked T. brasiliensis nymphs (p<0.01), and Cd-marked R. nasutus nymphs (p<0.01) had significantly shorter lifespan. Conclusions/Significance Previous evidence shows that there is little or no genetic differentiation between populations at the microgeographic level, which often precludes indirect estimations of dispersal capability based on genetic markers. In such situations, MRR studies are more suitable as they measure insect movement directly from one site to another, instead of effective migration (i.e. gene flow). The determination of a reliable and persistent marking method is the first step towards the development of meaningful ecological estimates through the application of MRR methodology. Here, we have identified trace elements that can be used for mark and recapture studies of three triatomine species in Brazil.

Bionomia de Culex quinquefasciatus em áreas urbanas no Rio de Janeiro, RJ

OBJECTIVE To evaluate density, parity rates, daily survival and longevity of natural populations of Culex quinquefasciatus in three neighborhoods with distinct socio-economic and infrastructure profiles. METHODS Mosquito collections of the Culex quinquefasciatus species were performed weekly during two four month periods, from August to November 2008 (spring) and March to June 2009 (fall), in a favela (slum), a suburban area and a middle class area of Rio de Janeiro, Southeastern Brazil. Collections were performed with backpack aspirators, in 20 randomly selected houses in each area per week, during 15-20 minutes per house. Ovaries were removed from captured females and classified as initial, intermediary or final stage. Furthermore, females were dissected for determination of parity based on the condition of the tracheal system. Mosquito survival rate and longevity were estimated on a per month basis for each neighborhood. RESULTS We collected a total of 2,062 Culex quinquefasciatus, but monthly vector density was not correlated with temperature and rainfall. We dissected the ovaries of 625 Culex quinquefasciatus, and overall, there was a higher proportion of nulliparous females during the dryer months, while gravid females were more frequent in rainy months. In the middle class neighborhood, the parity rate reached up to 93.75% with survivorship of 0.979. Lower parity and survival rates were obtained in the suburban area (as low as 36.4% parity and 0.711 daily survival). Up to 84.7% of Culex quinquefasciatus females could survive the eight day period needed to complete West Nile Virus incubation. CONCLUSIONS The survival rate of Culex quinquefasciatus varied significantly between the neighborhoods. This suggests that vectorial capacity and disease transmission risk may vary greatly between different urban areas, which is potentially useful information for vector control programs.OBJETIVO: Avaliar densidade, taxa de paridade, sobrevivencia diaria e longevidade de populacoes naturais de Culex quinquefasciatus de tres localidades com diferentes perfis socioeconomicos e de infraestrutura. METODOS: Mosquitos da especie Culex quinquefasciatus foram coletados semanalmente em dois periodos de quatro meses, de agosto a novembro de 2008 e de marco a junho de 2009, em uma comunidade, uma area de suburbio e um bairro de classe media no Rio de Janeiro, RJ. As coletas foram realizadas com o auxilio de aspiradores costais em 20 domicilios, aleatoriamente selecionados por semana e por area, durante 15-20 minutos por domicilio. As femeas capturadas tiveram os ovarios removidos e classificados em estagios inicial, intermediario e final. Adicionalmente, as femeas foram dissecadas para determinacao da paridade de acordo com as condicoes do sistema traqueal. A taxa de sobrevivencia e a longevidade das femeas foram estimadas por mes, para cada localidade. RESULTADOS: Foram coletados 2.062 Culex quinquefasciatus, porem a densidade mensal deste vetor nao apresentou correlacao com temperatura e precipitacao. Dissecamos os ovarios de 625 Culex quinquefasciatus e obtivemos maior proporcao de femeas nuliparas durante os meses mais secos, enquanto femeas gravidas foram mais frequentes nos meses chuvosos. A taxa de paridade foi de ate 93,75% no bairro de classe media, com sobrevivencia de 0,979. Menores valores de paridade e sobrevivencia foram obtidos no suburbio (comecando em 36,4% de paridade e 0,711 de sobrevivencia diaria). Ate 84,7% das femeas de Culex quinquefasciatus poderiam sobreviver ao periodo de oito dias, necessario para completar o periodo de incubacao do virus da febre do Nilo Ocidental. CONCLUSOES: A taxa de sobrevivencia de Culex quinquefasciatus variou significativamente entre os bairros, o que sugere que a capacidade vetorial e o risco de transmissao de doencas podem variar entre diferentes areas urbanas, informacao relevante para o planejamento dos programas de controle de vetores.