Comparison of Vector Competence of Aedes mediovittatus and Aedes aegypti for Dengue Virus: Implications for Dengue Control in the Caribbean

Background

Aedes mediovittatus mosquitoes are found throughout the Greater Antilles in the Caribbean and often share the same larval habitats with Ae. Aegypti, the primary vector for dengue virus (DENV). Implementation of vector control measures to control dengue that specifically target Ae. Aegypti may not control DENV transmission in Puerto Rico (PR). Even if Ae. Aegypti is eliminated or DENV refractory mosquitoes are released, DENV transmission may not cease when other competent mosquito species like Ae. Mediovittatus are present. To compare vector competence of Ae. Mediovittatus and Ae. Aegypti mosquitoes, we studied relative infection and transmission rates for all four DENV serotypes.

Methods

To compare the vector competence of Ae. Mediovittatus and Ae. Aegypti, mosquitoes were exposed to DENV 1–4 per os at viral titers of 5–6 logs plaque-forming unit (pfu) equivalents. At 14 days post infectious bloodmeal, viral RNA was extracted and tested by qRT-PCR to determine infection and transmission rates. Infection and transmission rates were analyzed with a generalized linear model assuming a binomial distribution.

Results

Ae. Aegypti had significantly higher DENV-4 infection and transmission rates than Ae. mediovittatus.

Conclusions

This study determined that Ae. Mediovittatus is a competent DENV vector. Therefore dengue prevention programs in PR and the Caribbean should consider both Ae. Mediovittatus and Ae. Aegypti mosquitoes in their vector control programs.     

Susceptibility of Florida Aedes aegypti and Aedes albopictus to dengue viruses from Puerto Rico

Locally acquired dengue cases in the continental U.S. are rare. However, outbreaks of dengue‐1 during 2009, 2010, and 2013 in Florida and dengue‐1 and −2 in Texas suggest vulnerability to transmission. Travel and commerce between Puerto Rico and the U.S. mainland is common, which may pose a risk for traveler‐imported dengue cases. Mosquitoes were collected in Florida and used to evaluate their susceptibility to dengue viruses (DENV) from Puerto Rico. Aedes aegypti and Ae. albopictus were susceptible to virus infection with DENV‐1 and −2. No significant differences were observed in rates of midgut infection or dissemination between Ae. aegypti or Ae. albopictus for DENV‐1 (6–14%). Aedes aegypti was significantly more susceptible to midgut infection with DENV‐2 than Ae. albopictus (Ae. aegypti, ∼28%; Ae. albopictus, ∼9%). The dissemination rate with dengue‐2 virus for Ae. aegypti (23%) was greater than Ae. albopictus (0%), suggesting that Ae. albopictus is not likely to be an important transmitter of the DENV‐2 isolate from Puerto Rico. These results are discussed in light of Florida's vulnerability to DENV transmission.     

Local and Global Effects of Climate on Dengue Transmission in Puerto Rico

The four dengue viruses, the agents of dengue fever and dengue hemorrhagic fever in humans, are transmitted predominantly by the mosquito Aedes aegypti. The abundance and the transmission potential of Ae. aegypti are influenced by temperature and precipitation. While there is strong biological evidence for these effects, empirical studies of the relationship between climate and dengue incidence in human populations are potentially confounded by seasonal covariation and spatial heterogeneity. Using 20 years of data and a statistical approach to control for seasonality, we show a positive and statistically significant association between monthly changes in temperature and precipitation and monthly changes in dengue transmission in Puerto Rico. We also found that the strength of this association varies spatially, that this variation is associated with differences in local climate, and that this relationship is consistent with laboratory studies of the impacts of these factors on vector survival and viral replication. These results suggest the importance of temperature and precipitation in the transmission of dengue viruses and suggest a reason for their spatial heterogeneity. Thus, while dengue transmission may have a general system, its manifestation on a local scale may differ from global expectations.     

Atmospheric control of Aedes aegypti populations in Buenos Aires (Argentina) and its variability

The mosquito Aedes aegypti is the main urban vector responsible for the transmission of dengue fever and dengue hemorrhagic fever. The city of Buenos Aires, Argentina, is located at the southern end of the world distribution of the species. The population abundance of Ae. aegypti is mainly regulated by environmental factors. We calculated the potential number of times that a female could lay eggs during its mean life expectancy, based on potential egg production and daily meteorological records. The model considers those variables implying physical hazard to the survival of Ae. aegypti, mosquito flying activity and oviposition. The results, obtained after calibration and validation of the model with field observations, show significant correlation (P<0.001) for different lags depending on the life stage. From these results, more favorable atmospheric conditions for Ae. aegypti reproduction (linked to the urban climatic change) can be observed. The climatic variability in the last decade resembles conditions at the end of 19th century. Received: 3 March 1999 / Revised: 24 November 1999 / Accepted: 24 January 2000     

A comparison of mosquito densities,weather and infection rates of Aedes aegypti during the first epidemics of Chikungunya (2014) and Zika (2016) in areas with and without vector control in Puerto Rico

In Puerto Rico, the first records of the transmission of Chikungunya (CHIKV) and Zika (ZIKV) viruses were confirmed in May 2014 and December 2015, respectively. Transmission of CHIKV peaked in September 2014, whereas that of ZIKV peaked in August 2016. The emergence of these mosquito‐transmitted arboviruses in the context of a lack of human population immunity allowed observations of whether the outbreaks were associated with Aedes aegypti (Diptera: Culicidae) densities and weather. Mosquito density was monitored weekly in four communities using sentinel autocidal gravid ovitraps (AGO traps) during 2016 in order to provide data to be compared with the findings of a previous study carried out during the 2014 CHIKV epidemic. Findings in two communities protected against Ae. aegypti using mass AGO trapping (three traps per house in most houses) were compared with those in two nearby communities without vector control. Mosquito pools were collected to detect viral RNA of ZIKV, CHIKV and dengue virus. In areas without vector control, mosquito densities and rates of ZIKV detection in 2016 were significantly higher, similarly to those observed for CHIKV in 2014. The density of Ae. aegypti in treated sites was less than two females/trap/week, which is similar to the putative adult female threshold for CHIKV transmission. No significant differences in mosquito density or infection rates with ZIKV and CHIKV at the same sites between years were observed. Although 2016 was significantly wetter, mosquito densities were similar.     

Dengue Virus Type 2 Infections of Aedes aegypti Are Modulated by the Mosquito's RNA Interference Pathway

A number of studies have shown that both innate and adaptive immune defense mechanisms greatly influence the course of human dengue virus (DENV) infections, but little is known about the innate immune response of the mosquito vector Aedes aegypti to arbovirus infection. We present evidence here that a major component of the mosquito innate immune response, RNA interference (RNAi), is an important modulator of mosquito infections. The RNAi response is triggered by double-stranded RNA (dsRNA), which occurs in the cytoplasm as a result of positive-sense RNA virus infection, leading to production of small interfering RNAs (siRNAs). These siRNAs are instrumental in degradation of viral mRNA with sequence homology to the dsRNA trigger and thereby inhibition of virus replication. We show that although dengue virus type 2 (DENV2) infection of Ae. aegypti cultured cells and oral infection of adult mosquitoes generated dsRNA and production of DENV2-specific siRNAs, virus replication and release of infectious virus persisted, suggesting viral circumvention of RNAi. We also show that DENV2 does not completely evade RNAi, since impairing the pathway by silencing expression of dcr2, r2d2, or ago2, genes encoding important sensor and effector proteins in the RNAi pathway, increased virus replication in the vector and decreased the extrinsic incubation period required for virus transmission. Our findings indicate a major role for RNAi as a determinant of DENV transmission by Ae. aegypti.     

Satellite observation to assess dengue risk due to Aedes aegypti and Aedes albopictus in a subtropical city of Argentina

Earth observation environmental features measured through remote sensing and models of vector mosquitoes species Aedes aegypti and Ae. albopictus provide an advancement with regards to dengue risk in urban environments of subtropical areas of Argentina. The authors aim to estimate the effect of landscape coverage and spectral indices (Normalized Difference Vegetation Index [NDVI], Normalized Difference Water Index [NDWI] and Normalized Difference Built-up Index [NDBI]) on the larvae abundance of Ae. aegypti and Ae. albopictus in Eldorado, Misiones, Argentina using remote satellite sensors. Larvae of these species were collected monthly (June 2016 to April 2018), in four environments: tire repair shops, cemeteries, dwellings and an urban natural park. The proportion of landscape coverage (water, urban areas, bare soil, low vegetation and high vegetation) was determined from the supervised classification of Sentinel-2 images and spectral indices, calculated. The authors developed spatial models of both vector species by generalized linear mixed models. The model's results showed that Ae. aegypti larvae abundance was better modelled by NDVI minimum values, NDBI maximum values and the interaction between them. For Ae. albopictus proportion of bare soil, low vegetation and the interaction between both variables explained better the abundance.     

Chronology of sodium channel mutations associated with pyrethroid resistance in Aedes aegypti

Aedes aegypti is the primary mosquito vector of dengue, yellow fever, Zika and chikungunya. Current strategies to control Ae. aegypti rely heavily on insecticide interventions. Pyrethroids are a major class of insecticides used for mosquito control because of their fast acting, highly insecticidal activities and low mammalian toxicity. However, Ae. aegypti populations around the world have begun to develop resistance to pyrethroids. So far, more than a dozen mutations in the sodium channel gene have been reported to be associated with pyrethroid resistance in Ae. aegypti. Co-occurrence of resistance-associated mutations is common in pyrethroid-resistant Ae. aegypti populations. As global use of pyrethroids in mosquito control continues, new pyrethroid-resistant mutations keep emerging. In this microreview, we compile pyrethroid resistance-associated mutations in Ae. aegypti in a chronological order, as they were reported, and summarize findings from functional evaluation of these mutations in an in vitro sodium channel expression system. We hope that the information will be useful for tracing possible evolution of pyrethroid resistance in this important human disease vector, in addition to the development of methods for global monitoring and management of pyrethroid resistance in Ae. aegypti.     

Elimination of Aedes aegypti in northern Australia, 2004–2006

The Northern Territory (NT) of Australia is currently free of the dengue mosquito Aedes (Stegomyia) aegypti (L). However, on 17 February 2004, two Ae. aegypti adults were captured in two routine CO₂‐baited encephalitis virus surveillance traps in Tennant Creek, located 990 km south of Darwin in the NT. The detection triggered an immediate survey and control response undertaken by the NT Department of Health and Community Services, followed by a Commonwealth of Australia‐funded Ae. aegypti elimination program. This report details the methods and results of the detection and subsequent elimination activities that were carried out between 2004 and 2006, returning the NT to its dengue vector‐free status. There have been very few successful Ae. aegypti elimination programs in the world. This purposeful mosquito elimination for Australia was officially declared on 5 April 2006.     

State‐wide survey of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Florida

Aedes aegypti and Aedes albopictus are invasive mosquito species with geographic ranges that have oscillated within Florida since their presence was first documented. Local transmission of dengue, chikungunya, and Zika viruses serves as evidence of the public health importance of these two species. It is important to have detailed knowledge of their distribution to aid in mosquito control efforts and understand the risk of arbovirus transmission to humans. Through a partnership involving the University of Florida Institute of Food and Agricultural Sciences Cooperative Extension Service and the Florida Medical Entomology Laboratory; the Florida Department of Health; and mosquito control agencies throughout Florida, a container mosquito surveillance program involving all life stages was launched in the summer of 2016 to detect the presence of Ae. aegypti and Ae. albopictus. Results from this survey were mapped to provide a picture of the current known distribution of Ae. aegypti and Ae. albopictus in Florida. Aedes aegypti and/or Ae. albopictus were detected in the 56 counties that were part of the survey. Only Aedes albopictus was detected in 26 counties, primarily in the panhandle region of Florida. The results of this work underscore the importance of maintaining container mosquito surveillance in a state where chikungunya, dengue, and Zika viruses are present and where there is continued risk for exotic arbovirus introductions.     

Septic tanks as larval habitats for the mosquitoes Aedes aegypti and Culex quinquefasciatus in Playa‐Playita,Puerto Rico

Adult Aedes aegypti (Linnaeus) (Diptera: Culicidae) were previously recovered from emergence traps on septic tanks in southeastern Puerto Rico. In this study we quantified immature mosquito abundance and its relationship with structural variables of the septic tanks and chemical properties of the water containing raw sewage. A miniaturized floating funnel trap was used to sample 89 septic tanks for larvae in the Puerto Rican community of Playa‐Playita. Aedes aegypti larvae were recovered from 18% of the sampled tanks (10.3 larvae per septic tank per day). Larval presence was positively associated with cracking of the septic tank walls and uncovered access ports. Larval abundance was positively associated with cracking of the septic tank walls and larger tank surface areas, and inversely associated with the total dissolved solids (TDS). Culex quinquefasciatus (Say) larvae were also recovered from 74% of the septic tanks (129.6 larvae per septic tank per day). Larval presence was negatively associated with TDS in the water and larval abundance was positively associated with cracking of the septic tank walls. A screened, plastic emergence trap was used to sample 93 septic tanks within the community for Ae. aegypti and Cx. quinquefasciatus adults. Aedes aegypti adults were recovered from 49% of the sampled tanks (8.7 adults per septic tank per day) and Cx. quinquefasciatus adults were recovered from 97% of the sampled tanks (155.5 adults per septic tank per day). Aedes aegypti adult presence was positively associated with cracking, uncapped openings and septic water pH. The Ae. aegypti adult counts were positively associated with cracking and inversely associated with TDS and conductivity. This study marks the first published record of the recovery of Ae. aegypti larvae from holding tanks containing raw sewage in the Caribbean region. Our study indicates that Ae. aegypti larvae are present in sewage water and that septic tanks have at least the potential to maintain dengue transmission during the dry season.     

A Virulent Wolbachia Infection Decreases the Viability of the Dengue Vector Aedes aegypti during Periods of Embryonic Quiescence

A new approach for dengue control has been proposed that relies on life-shortening strains of the obligate intracellular bacterium Wolbachia pipientis to modify mosquito population age structure and reduce pathogen transmission. Previously we reported the stable transinfection of the major dengue vector Aedes aegypti with a life-shortening Wolbachia strain (wMelPop-CLA) from the vinegar fly Drosophila melanogaster. Here, we report a further characterization of the phenotypic effects of this virulent Wolbachia infection on several life-history traits of Ae. aegypti. Minor costs of wMelPop-CLA infection for pre-imaginal survivorship, development and adult size were found. However, we discovered that the wMelPop-CLA infection dramatically decreased the viability of desiccated Ae. aegypti eggs over time. Similarly, the reproductive fitness of wMelPop-CLA infected Ae. aegypti females declined with age. These results reveal a general pattern associated with wMelPop-CLA induced pathogenesis in this mosquito species, where host fitness costs increase during aging of both immature and adult life-history stages. In addition to influencing the invasion dynamics of this particular Wolbachia strain, we suggest that the negative impact of wMelPop-CLA on embryonic quiescence may have applied utility as a tool to reduce mosquito population size in regions with pronounced dry seasons or in regions that experience cool winters.     

Prospective field study of transovarial dengue‐virus transmission by two different forms of Aedes aegypti in an urban area of Bangkok,Thailand

A prospective field study was conducted to determine transovarial dengue‐virus transmission in two forms of Aedes aegypti mosquitoes in an urban district of Bangkok, Thailand. Immature Aedes mosquitoes were collected monthly for one year and reared continuously until adulthood in the laboratory. Mosquitoes assayed for dengue virus were processed in pools and their dengue virus infection status was determined by one‐step RT‐PCR and nested‐PCR methods. Of a total 15,457 newly emerged adult Ae. aegypti, 98.2% were dark and 1.8% of the pale form. The results showed that the minimum infection rate (MIR) by transovarial transmission (TOT) of dengue virus during the one‐year study ranged between 0 to 24.4/1,000 mosquitoes. Dengue virus TOT increased gradually during the hot summer months, reaching a peak in April‐June, while dengue cases peaked in September, a rainy month near the end of the rainy season. Therefore, mosquito infections due to TOT were prevalent four months before a high incidence of human infections. TOT dengue virus infections occurred in both forms of Ae. aegypti. All four dengue serotypes were detected, with DEN‐4 predominant, followed by DEN‐3, DEN‐1, and DEN‐2, respectively.     

The Neovolcanic Axis Is a Barrier to Gene Flow among Aedes aegypti Populations in Mexico That Differ in Vector Competence for Dengue 2 Virus

Background

Aedes aegypti is the main mosquito vector of the four serotypes of dengue virus (DENV). Previous population genetic and vector competence studies have demonstrated substantial genetic structure and major differences in the ability to transmit dengue viruses in Ae. aegypti populations in Mexico.

Methodology/Principal Findings

Population genetic studies revealed that the intersection of the Neovolcanic axis (NVA) with the Gulf of Mexico coast in the state of Veracruz acts as a discrete barrier to gene flow among Ae. aegypti populations north and south of the NVA. The mosquito populations north and south of the NVA also differed in their vector competence (VC) for dengue serotype 2 virus (DENV2). The average VC rate for Ae. aegypti mosquitoes from populations from north of the NVA was 0.55; in contrast the average VC rate for mosquitoes from populations from south of the NVA was 0.20. Most of this variation was attributable to a midgut infection and escape barriers. In Ae. aegypti north of the NVA 21.5% failed to develop midgut infections and 30.3% of those with an infected midgut failed to develop a disseminated infection. In contrast, south of the NVA 45.2% failed to develop midgut infections and 62.8% of those with an infected midgut failed to develop a disseminated infection.

Conclusions

Barriers to gene flow in vector populations may also impact the frequency of genes that condition continuous and epidemiologically relevant traits such as vector competence. Further studies are warranted to determine why the NVA is a barrier to gene flow and to determine whether the differences in vector competence seen north and south of the NVA are stable and epidemiologically significant.     

Nonlinear impacts of climatic variability on the density‐dependent regulation of an insect vector of disease

Aedes aegypti is one of the most common urban tropical mosquito species and an important vector of dengue, chikungunya, and yellow fever viruses. It is also an organism with a complex life history where larval stages are aquatic and adults are terrestrial. This ontogenetic niche shift could shape the density‐dependent regulation of this and other mosquito species, because events that occur during the larval stages impact adult densities. Herein, we present results from simple density‐dependent mathematical models fitted using maximum likelihood methods to weekly time series data from Puerto Rico and Thailand. Density‐dependent regulation was strong in both populations. Analysis of climatic forcing indicated that populations were more sensitive to climatic variables with low kurtosis, i.e., climatic factors highly variable around the median, rainfall in Puerto Rico, and temperature in Thailand. Changes in environmental variability appear to drive sharp changes in the abundance of mosquitoes. The identification of density‐independent (i.e., exogenous) variables forcing sharp changes in disease vector populations using the exogenous factors statistical properties, such as kurtosis, could be useful to assess the impacts of changing climate patterns on the transmission of vector‐borne diseases.     

Australia's Dengue Risk Driven by Human Adaptation to Climate Change

Background

The reduced rainfall in southeast Australia has placed this region''s urban and rural communities on escalating water restrictions, with anthropogenic climate change forecasts suggesting that this drying trend will continue. To mitigate the stress this may place on domestic water supply, governments have encouraged the installation of large domestic water tanks in towns and cities throughout this region. These prospective stable mosquito larval sites create the possibility of the reintroduction of Ae. aegypti from Queensland, where it remains endemic, back into New South Wales and other populated centres in Australia, along with the associated emerging and re-emerging dengue risk if the virus was to be introduced.

Methodology/Principal Findings

Having collated the known distribution of Ae. aegypti in Australia, we built distributional models using a genetic algorithm to project Ae. aegypti''s distribution under today''s climate and under climate change scenarios for 2030 and 2050 and compared the outputs to published theoretical temperature limits. Incongruence identified between the models and theoretical temperature limits highlighted the difficulty of using point occurrence data to study a species whose distribution is mediated more by human activity than by climate. Synthesis of this data with dengue transmission climate limits in Australia derived from historical dengue epidemics suggested that a proliferation of domestic water storage tanks in Australia could result in another range expansion of Ae. aegypti which would present a risk of dengue transmission in most major cities during their warm summer months.

Conclusions/Significance

In the debate of the role climate change will play in the future range of dengue in Australia, we conclude that the increased risk of an Ae. aegypti range expansion in Australia would be due not directly to climate change but rather to human adaptation to the current and forecasted regional drying through the installation of large domestic water storing containers. The expansion of this efficient dengue vector presents both an emerging and re-emerging disease risk to Australia. Therefore, if the installation and maintenance of domestic water storage tanks is not tightly controlled, Ae. aegypti could expand its range again and cohabit with the majority of Australia''s population, presenting a high potential dengue transmission risk during our warm summers.     

Aedes aegypti lachesin protein binds to the domain III of envelop protein of Dengue virus‐2 and inhibits viral replication

Dengue virus (DENV) comprises of four serotypes (DENV‐1 to ‐4) and is medically one of the most important arboviruses (arthropod‐borne virus). DENV infection is a major human health burden and is transmitted between humans by the insect vector, Aedes aegypti. Ae. aegypti ingests DENV while feeding on infected humans, which traverses through its gut, haemolymph and salivary glands of the mosquito before being injected into a healthy human. During this process of transmission, DENV must interact with many proteins of the insect vector, which are important for its successful transmission. Our study focused on the identification and characterisation of interacting protein partners in Ae. aegypti to DENV. Since domain III (DIII) of envelope protein (E) is exposed on the virion surface and is involved in virus entry into various cells, we performed phage display library screening against domain III of the envelope protein (EDIII) of DENV‐2. A peptide sequence showing similarity to lachesin protein was found interacting with EDIII. The lachesin protein was cloned, heterologously expressed, purified and used for in vitro interaction studies. Lachesin protein interacted with EDIII and also with DENV. Further, lachesin protein was localised in neuronal cells of different organs of Ae. aegypti by confocal microscopy. Blocking of lachesin protein in Ae. aegypti with anti‐lachesin antibody resulted in a significant reduction in DENV replication.     

Population dynamics of Aedes aegypti and dengue as influenced by weather and human behavior in San Juan, Puerto Rico

Previous studies on the influence of weather on Aedes aegypti dynamics in Puerto Rico suggested that rainfall was a significant driver of immature mosquito populations and dengue incidence, but mostly in the drier areas of the island. We conducted a longitudinal study of Ae. aegypti in two neighborhoods of the metropolitan area of San Juan city, Puerto Rico where rainfall is more uniformly distributed throughout the year. We assessed the impacts of rainfall, temperature, and human activities on the temporal dynamics of adult Ae. aegypti and oviposition. Changes in adult mosquitoes were monitored with BG-Sentinel traps and oviposition activity with CDC enhanced ovitraps. Pupal surveys were conducted during the drier and wetter parts of the year in both neighborhoods to determine the contribution of humans and rains to mosquito production. Mosquito dynamics in each neighborhood was compared with dengue incidence in their respective municipalities during the study. Our results showed that: 1. Most pupae were produced in containers managed by people, which explains the prevalence of adult mosquitoes at times when rainfall was scant; 2. Water meters were documented for the first time as productive habitats for Ae. aegypti; 3. Even though Puerto Rico has a reliable supply of tap water and an active tire recycling program, water storage containers and discarded tires were important mosquito producers; 4. Peaks in mosquito density preceded maximum dengue incidence; and 5. Ae. aegypti dynamics were driven by weather and human activity and oviposition was significantly correlated with dengue incidence.