Gonotrophic Cycle Research Articles

Effective disposal of nitrogen waste in blood-fed Aedes aegypti mosquitoes requires alanine aminotransferase.

To better understand the mechanisms responsible for the success of female mosquitoes in their disposal of excess nitrogen, we investigated the role of alanine aminotransferase (ALAT) in blood-fed Aedes aegypti. Transcript and protein levels from the 2 ALAT genes were analyzed in sucrose- and blood-fed A. aegypti tissues. ALAT1 and ALAT2 exhibit distinct expression patterns in tissues during the first gonotrophic cycle. Injection of female mosquitoes with either double-stranded RNA (dsRNA)-ALAT1 or dsRNA ALAT2 significantly decreased mRNA and protein levels of ALAT1 or ALAT2 in fat body, thorax, and Malpighian tubules compared with dsRNA firefly luciferase-injected control mosquitoes. The silencing of either A. aegypti ALAT1 or ALAT2 caused unexpected phenotypes such as a delay in blood digestion, a massive accumulation of uric acid in the midgut posterior region, and a significant decrease of nitrogen waste excretion during the first 48 h after blood feeding. Concurrently, the expression of genes encoding xanthine dehydrogenase and ammonia transporter (Rhesus 50 glycoprotein) were significantly increased in tissues of both ALAT1- and ALAT2-deficient females. Moreover, perturbation of ALAT1 and ALAT2 in the female mosquitoes delayed oviposition and reduced egg production. These novel findings underscore the efficient mechanisms that blood-fed mosquitoes use to avoid ammonia toxicity and free radical damage.-Mazzalupo, S., Isoe, J., Belloni, V., Scaraffia, P. Y. Effective disposal of nitrogen waste in blood-fed Aedes aegypti mosquitoes requires alanine aminotransferase.

Parity and longevity of Aedes aegypti according to temperatures in controlled conditions and consequences on dengue transmission risks.

BackgroundIn Guadeloupe, Aedes aegypti mosquitoes are the only vectors of dengue and chikungunya viruses. For both diseases, vector control is the only tool for preventing epidemics since no vaccine or specific treatment is available. However, to efficiently implement control of mosquitoes vectors, a reliable estimation of the transmission risks is necessary. To become infective an Ae. aegypti female must ingest the virus during a blood meal and will not be able to transmit the virus during another blood-meal until the extrinsic incubation period is completed. Consequently the aged females will carry more infectious risks. The objectives of the present study were to estimate under controlled conditions the expectation of infective life for females and thus the transmission risks in relation with their reproductive cycle and parity status.Methodology/Principal FindingsLarvae of Ae. aegypti were collected in central Guadeloupe and breed under laboratory conditions until adult emergence. The experiments were performed at constant temperatures (± 1.5°C) of 24°C, 27°C and 30°C on adults females from first generation (F1). Females were kept and fed individually and records of blood-feeding, egg-laying and survival were done daily. Some females were dissected at different physiological stages to observe the ovaries development. The data were analyzed to follow the evolution of parity rates, the number of gonotrophic cycles, the fecundity and to study the mean expectation of life and the mean expectation of infective life for Ae. aegypti females according to temperatures. The expectation of life varies with the parity rates and according to the temperatures, with durations from about 10 days at low parity rates at the higher temperature to an optimal duration of about 35 days when 70% of females are parous at 27°C. Infective life expectancy was found highly variable in the lower parous rates and again the optimal durations were found when more than 50% of females are parous for the mean temperatures of 27°C and 30°C.ConclusionParity rates can be determined for field collected females and could be a good proxy of the expectation of infective life according to temperatures. However, for the same parity rates, the estimation of infective life expectation is very different between Ae. aegypti and Anopheles gambiae mosquitoes. Correlation of field parity rates with transmission risks requires absolutely to be based on Ae. aegypti models, since available Anopheles sp. models underestimate greatly the females longevity.

Indoor use of attractive toxic sugar bait (ATSB) to effectively control malaria vectors in Mali, West Africa.

BackgroundAttractive toxic sugar bait (ATSB) solutions containing any gut toxins can be either sprayed on plants or used in simple bait stations to attract and kill sugar-feeding female and male mosquitoes. This field study in Mali demonstrates the effect of ATSB bait stations inside houses as a vector control method that targets and kills endophilic African malaria vectors.MethodsThe studies were conducted in five villages located near the River Niger, Mali. Baseline village-wide assessments of densities for female and male Anopheles gambiae sensu lato were performed by pyrethrum spray collections (PSC) in ten houses in each of five villages. To determine the rate of mosquito feeding on bait stations, one bait station per house containing attractive sugar bait (ASB) (without toxin) plus a food dye marker, was set up in ten houses in each of the five villages. PSC collections were conducted on the following day and the percentage of female and male mosquitoes that had fed was determined by visual inspection for the dye marker. Then, a 50-day field trial was done. In an experimental village, one bait station containing ATSB (1% boric acid active ingredient) was placed per bedroom (58 bedrooms), and indoor densities of female and male An. gambiae s.l. were subsequently determined by PSC, and female mosquitoes were age graded.ResultsIn the five villages, the percentages of An. gambiae s.l. feeding inside houses on the non-toxic bait stations ranged from 28.3 to 53.1% for females and 36.9 to 78.3% for males. Following ATSB indoor bait station presentation, there was a significant reduction, 90% in female and 93% in male populations, of An. gambiae s.l. at the experimental village. A 3.8-fold decrease in the proportion of females that had undergone four or more gonotrophic cycles was recorded at the experimental village, compared to a 1.2-fold increase at the control village.ConclusionThe field trial demonstrates that An. gambiae s.l. feed readily from ATSB bait stations situated indoors, leading to a substantial reduction in the proportion of older female mosquitoes. This study demonstrates that ATSB inside houses can achieve impressive malaria vector control in Africa.

Predicting Scenarios for Successful Autodissemination of Pyriproxyfen by Malaria Vectors from Their Resting Sites to Aquatic Habitats; Description and Simulation Analysis of a Field-Parameterizable Model.

BackgroundLarge-cage experiments indicate pyriproxifen (PPF) can be transferred from resting sites to aquatic habitats by Anopheles arabiensis - malaria vector mosquitoes to inhibit emergence of their own offspring. PPF coverage is amplified twice: (1) partial coverage of resting sites with PPF contamination results in far higher contamination coverage of adult mosquitoes because they are mobile and use numerous resting sites per gonotrophic cycle, and (2) even greater contamination coverage of aquatic habitats results from accumulation of PPF from multiple oviposition events.Methods and FindingsDeterministic mathematical models are described that use only field-measurable input parameters and capture the biological processes that mediate PPF autodissemination. Recent successes in large cages can be rationalized, and the plausibility of success under full field conditions can be evaluated a priori. The model also defines measurable properties of PPF delivery prototypes that may be optimized under controlled experimental conditions to maximize chances of success in full field trials. The most obvious flaw in this model is the endogenous relationship that inevitably occurs between the larval habitat coverage and the measured rate of oviposition into those habitats if the target mosquito species is used to mediate PPF transfer. However, this inconsistency also illustrates the potential advantages of using a different, non-target mosquito species for contamination at selected resting sites that shares the same aquatic habitats as the primary target. For autodissemination interventions to eliminate malaria transmission or vector populations during the dry season window of opportunity will require comprehensive contamination of the most challenging subset of aquatic habitats that persist or retain PPF activity (U x) for only one week , where U x = 7 days). To achieve >99% contamination coverage of these habitats will necessitate values for the product of the proportional coverage of the ovipositing mosquito population with PPF contamination (C M) by the ovitrap-detectable rates of oviposition by wild mosquitoes into this subset of habitats , divided by the titre of contaminated mosquitoes required to render them unproductive , that approximately approach unity .ConclusionsThe simple multiplicative relationship between C M and , and the simple exponential decay effect they have upon uncontaminated aquatic habitats, allows application of this model by theoreticians and field biologists alike.

Potential effects of warmer worms and vectors on onchocerciasis transmission in West Africa.

Development times of eggs, larvae and pupae of vectors of onchocerciasis (Simulium spp.) and of Onchocerca volvulus larvae within the adult females of the vectors decrease with increasing temperature. At and above 25°C, the parasite could reach its infective stage in less than 7 days when vectors could transmit after only two gonotrophic cycles. After incorporating exponential functions for vector development into a novel blackfly population model, it was predicted that fly numbers in Liberia and Ghana would peak at air temperatures of 29°C and 34°C, about 3°C and 7°C above current monthly averages, respectively; parous rates of forest flies (Liberia) would peak at 29°C and of savannah flies (Ghana) at 30°C. Small temperature increases (less than 2°C) might lead to changes in geographical distributions of different vector taxa. When the new model was linked to an existing framework for the population dynamics of onchocerciasis in humans and vectors, transmission rates and worm loads were projected to increase with temperature to at least 33°C. By contrast, analyses of field data on forest flies in Liberia and savannah flies in Ghana, in relation to regional climate change predictions, suggested, on the basis of simple regressions, that 13–41% decreases in fly numbers would be expected between the present and before 2040. Further research is needed to reconcile these conflicting conclusions.

Interactions between a fungal entomopathogen and malaria parasites within a mosquito vector.

BackgroundMosquitoes are becoming increasingly resistant to the chemical insecticides currently available for malaria vector control, spurring interest in alternative management tools. One promising technology is the use of fungal entomopathogens. Fungi have been shown to impact the potential for mosquitoes to transmit malaria by reducing mosquito longevity and altering behaviour associated with flight and host location. Additionally, fungi could impact the development of malaria parasites within the mosquito via competition for resources or effects on the mosquito immune system. This study evaluated whether co-infection or superinfection with the fungal entomopathogen Beauveria bassiana affected malaria infection progress in Anopheles stephensi mosquitoes.MethodsThe study used two parasite species to examine possible effects of fungal infection at different parasite development stages. First, the rodent malaria model Plasmodium yoelii was used to explore interactions at the oocyst stage. Plasmodium yoelii produces high oocyst densities in infected mosquitoes and thus was expected to maximize host immunological and resource demands. Second, fungal interactions with mature sporozoites were evaluated by infecting mosquitoes with the human malaria species Plasmodium falciparum, which is highly efficient at invading mosquito salivary glands.ResultsWith P. yoelii, there was no evidence that fungal co-infection (on the same day as the blood meal) or superinfection (during a subsequent gonotrophic cycle after parasite infection) affected the proportion of mosquitoes with oocysts, the number of oocysts per infected mosquito or the number of sporozoites per oocyst. Similarly, for P. falciparum, there was no evidence that fungal infection affected sporozoite prevalence. Furthermore, there was no impact of infection with either malaria species on fungal virulence as measured by mosquito survival time.ConclusionsThese results suggest that the impact of fungus on malaria control potential is limited to the well-established effects on mosquito survival and transmission behaviour. Direct or indirect interactions between fungus and malaria parasites within mosquitoes appear to have little additional influence.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-014-0526-x) contains supplementary material, which is available to authorized users.

Application of Polovodova's method for the determination of physiological age and relationship between the level of parity and infectivity of Plasmodium falciparum in Anopheles gambiae s.s, south-eastern Benin.

BackgroundPolovodova method based on counting follicular dilatations estimates the number of egg-laying in mosquitoes. However, some researchers doubt the reliability of this method because of the absence of multiple dilatations in vectors after many gonotrophic cycles. It is in this context of controversy that our study was carried out to evaluate the importance of follicular dilatations in the determination of parity levels in An. gambiae s.s. Moreover, the application of this method allowed us to clarify the evolution of vectors’ infectivity to P. falciparum according to their parity level.MethodsWe used two techniques to determine the parity level in An. gambiae s.s. We used two batches of wild strain mosquitoes reproduced after a known number of egg-laying in laboratory. The first batch was submitted to oil injection in the ovaries using a micropipette. In the same way, the classic technique of ovaries dilaceration (a technique based on the Polovodova method) was applied to the second batch. In order to assess relationship between parity level and mosquitoes’ infectivity, Polovodova method was applied on vectors collected on humans. Finally, Heads and thoraces of these vectors were individually analyzed for P. falciparum antigen detection using an ELISA assay.ResultsIn the first batch including 50 female mosquitoes “never laid”, 50 “laid once”, 50 “laid twice” and 48 “three times”, oil injection technique revealed 42 nulliparous, 44 uniparous, 46 biparous and 44 triparous respectively. Overall, Polovodova method was effective using oil injection technique (p > 0.05). On the other hand, in the second batch that has a similar number of laying to the first batch, the application of Polovodova method through classical technique of ovaries dilaceration was ineffective with multiparous females (p < 0.05). Moreover, probability of vector infectivity increased with the number of egg-laying (p < 0.0001).ConclusionOur results revealed that the Polovodova method is reliable for estimating the number of egg-laying in Anopheles gambiae s.s. using oil injection technique in the ovaries. The study has also showed an increased likelihood of infectivity in vectors according to the number of egg-laying.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0731-7) contains supplementary material, which is available to authorized users.

Host feeding patterns of mosquitoes in a rural malaria-endemic region in hainan island, china.

Malaria is endemic in Wangxia Village of Hainan Island. In this area little is known about the host seeking behavior and feeding habit of mosquitoes. Three sites representing the most common habitat types in the village were selected to study the host seeking behavior and feeding habit of mosquitoes. Of the total 9 species belonging to 4 genera (Armigeres, Culex, Aedes, and Anopheles) collected in Wangxia Village, Culex tritaeniorhynchus and Cx. pipiens quinquefasciatus were the most commonly collected species. Armigeres subalbatus and Anopheles sinensis were moderately common species. Blood meal analysis confirmed that Cx. tritaeniorhynchus and Cx. p. quinquefasciatus fed on multiple hosts, mainly poultry but occasionally other animals. Anopheles sinensis, a vector of malaria, fed predominately on cattle hosts, followed by humans. Anopheles maculatus and An. barbirostris fed on both humans and domestic animals. Our results indicate that most mosquitoes in this area preferred domestic animals over humans and showed a tendency to feed on multiple hosts within the same gonotrophic cycle. Therefore, the potential role of domestic animals in arbovirus transmission should be evaluated as part of a strategy for controlling mosquito-borne diseases in this region.

Capacity of mosquitoes to transmit malaria depends on larval environment.

BackgroundAdult traits of holometabolous insects such as reproduction and survival can be shaped by conditions experienced during larval development. These “carry-over” effects influence not only individual life history and fitness, but can also impact interactions between insect hosts and parasites. Despite this, the implications of larval conditions for the transmission of human, wildlife and plant diseases that are vectored by insects remain poorly understood.MethodsWe used Anopheles stephensi mosquitoes and the rodent malaria, Plasmodium yoelii yoelii, to investigate whether quality of larval habitat influenced vectorial capacity of adult mosquitoes. Larvae were reared under two dietary conditions; one group received a diet commonly used for colony maintenance (0.3 mg/individual/day of Tetrafin fish food) while the other group received a reduced food diet (0.1 mg/individual/day). Upon emergence, adults were provided an infectious blood feed. We assessed the effects of diet on a range of larval and adult traits including larval development times and survival, number of emerging adults, adult body size and survival, gonotrophic cycle length, and mating success. We also estimated the effects of larval diet on parasite infection rates and growth kinetics within the adult mosquitoes.ResultsLarval dietary regime affected larval survival and development, as well as size, reproductive success and survival of adult mosquitoes. Larval diet also affected the intensity of initial Plasmodium infection (oocyst stage) and parasite replication, but without differences in overall infection prevalence at either the oocyst or sporozoite stage.ConclusionsTogether, the combined effects led to a relative reduction in vectorial capacity (a measure of the transmission potential of a mosquito population) in the low food treatment of 70%. This study highlights the need to consider environmental variation at the larval stages to better understand transmission dynamics and control of vector-borne diseases.

Coordinated changes in JH biosynthesis and JH hemolymph titers in Aedes aegypti mosquitoes

Juvenile hormone III (JH) is synthesized by the corpora allata (CA) and plays a key role in mosquito development and reproduction. A decrease in JH titer during the last instar larvae allows pupation and metamorphosis to proceed. As the anti-metamorphic role of JH comes to an end, the CA of the late pupa once again synthesizes JH, which plays an essential role in orchestrating reproductive maturation. In spite of the importance of Aedes aegypti as a vector, a detailed study of the changes of JH hemolymph titers during the gonotrophic cycle has never been performed. In the present studies, using a high performance liquid chromatography coupled to a fluorescent detector (HPLC–FD) method, we measured changes in JH levels in the hemolymph of female mosquitoes during the pupal and adult stages. Our results revealed tightly concomitant changes in JH biosynthesis and JH hemolymph titers during the gonotrophic cycle of female mosquito. Feeding high sugar diets resulted in an increase of JH titers, and mating also modified JH titers in hemolymph. In addition these studies confirmed that JH titer in mosquitoes is fundamentally determined by the rate of biosynthesis in the CA.

An agent-based model of the population dynamics of Anopheles gambiae.

BackgroundAgent-based models (ABMs) have been used to model the behaviour of individual mosquitoes and other aspects of malaria. In this paper, a conceptual entomological model of the population dynamics of Anopheles gambiae and the agent-based implementations derived from it are described. Hypothetical vector control interventions (HVCIs) are implemented to target specific activities in the mosquito life cycle, and their impacts are evaluated.MethodsThe core model is described in terms of the complete An. gambiae mosquito life cycle. Primary features include the development and mortality rates in different aquatic and adult stages, the aquatic habitats and oviposition. The density- and age-dependent larval and adult mortality rates (vector senescence) allow the model to capture the age-dependent aspects of the mosquito biology. Details of hypothetical interventions are also described.ResultsResults show that with varying coverage and temperature ranges, the hypothetical interventions targeting the gonotrophic cycle stages produce higher impacts than the rest in reducing the potentially infectious female (PIF) mosquito populations, due to their multi-hour mortality impacts and their applicability at multiple gonotrophic cycles. Thus, these stages may be the most effective points of target for newly developed and novel interventions. A combined HVCI with low coverage can produce additive synergistic impacts and can be more effective than isolated HVCIs with comparatively higher coverages. It is emphasized that although the model described in this paper is designed specifically around the mosquito An. gambiae, it could effectively apply to many other major malaria vectors in the world (including the three most efficient nominal anopheline species An. gambiae, Anopheles coluzzii and Anopheles arabiensis) by incorporating a variety of factors (seasonality cycles, rainfall, humidity, etc.). Thus, the model can essentially be treated as a generic Anopheles model, offering an excellent framework for such extensions. The utility of the core model has also been demonstrated by several other applications, each of which investigates well-defined biological research questions across a variety of dimensions (including spatial models, insecticide resistance, and sterile insect techniques).

Regional variation of climatic influences on West Nile virus outbreaks in the United States.

The national resurgence of human West Nile virus (WNV) disease in 2012 raised questions about the factors responsible for WNV outbreaks. Interannual climatic variations may influence WNV amplification and transmission to humans through multiple pathways, including mosquito breeding habitats, gonotrophic cycles, extrinsic incubation, avian communities, and human behavior. We examined the influences of temperature and precipitation anomalies on interannual variation in human WNV cases in three regions of the United States. There were consistent positive influences of winter temperatures, weaker and more variable positive effects of spring and summer temperatures, and highly variable precipitation effects that ranged from positive to negative. The overwintering period may be a particularly important climatic constraint on the dynamics of WNV in cold-temperate regions of North America. Geographic differences in the seasonal timing and relative importance of climatic drivers of WNV risk likely reflect underlying variability in key ecological and social characteristics.

On a Reproductive Stage-Structured Model for the Population Dynamics of the Malaria Vector

A reproductive stage-structured deterministic differential equation model for the population dynamics of the human malaria vector is derived and analysed. The model captures the gonotrophic and behavioural life characteristics of the female Anopheles sp. mosquito and takes into consideration the fact that for the purposes of reproduction, the female Anopheles sp. mosquito must visit and bite humans (or animals) to harvest necessary proteins from blood that it needs for the development of its eggs. Focusing on mosquitoes that feed exclusively on humans, our results indicate the existence of a threshold parameter, the vectorial reproduction number, whose size increases with increasing number of gonotrophic cycles, and is also affected by the female mosquito's birth rate, its attraction and visitation rate to human residences, and its contact rate with humans. A stability analysis of the model indicates that the mosquito can establish itself in the environment if and only if the value of the vectorial reproduction number exceeds unity and that mosquito eradication is possible if the vectorial reproduction number is less than unity, since, then, the trivial steady state which always exist is unique and is globally and asymptotically stable. When a persistent vector population steady state exists, it is locally and asymptotically stable for a range of reproduction numbers, but can also be driven to instability via a Hopf bifurcation as the reproduction number increases further away from unity. The model derivation identifies and characterizes control parameters relating to activities such as human-mosquito contact and the mosquito's survival chances between blood meals and egg laying. Our results show that the total mosquito population size increases with increasing number of gonotrophic cycles. Therefore understanding the fundamental aspects of the mosquito's behaviour provides a pathway for the study of human-mosquito contact and mosquito population control. Control of the mosquito population densities would ultimately lead to malaria control.

Stability of the wMel Wolbachia Infection following invasion into Aedes aegypti populations.

The wMel infection of Drosophila melanogaster was successfully transferred into Aedes aegypti mosquitoes where it has the potential to suppress dengue and other arboviruses. The infection was subsequently spread into two natural populations at Yorkeys Knob and Gordonvale near Cairns, Queensland in 2011. Here we report on the stability of the infection following introduction and we characterize factors influencing the ongoing dynamics of the infection in these two populations. While the Wolbachia infection always remained high and near fixation in both locations, there was a persistent low frequency of uninfected mosquitoes. These uninfected mosquitoes showed weak spatial structure at both release sites although there was some clustering around two areas in Gordonvale. Infected females from both locations showed perfect maternal transmission consistent with patterns previously established pre-release in laboratory tests. After >2 years under field conditions, the infection continued to show complete cytoplasmic incompatibility across multiple gonotrophic cycles but persistent deleterious fitness effects, suggesting that host effects were stable over time. These results point to the stability of Wolbachia infections and their impact on hosts following local invasion, and also highlight the continued persistence of uninfected individuals at a low frequency most likely due to immigration.

Multi-agent modeling and simulation of Plasmodium falciparum dynamics considering sulfadoxine-pyrimethamine resistance and environmental factors

The appearance of resistant Plasmodium falciparum strains after the application of medication in human populations has generated global concern about the correct use of anti-malaria drugs. One of the challenges in malaria research is to find the correct methods to eradicate current strains while avoiding the appearance of new resistant strains. To recommend methods for malaria control, it is necessary to understand disease transmission and parasite dynamics and take into consideration spatial, temporal and environmental variations. In order to describe these elements, mathematical, stochastic and computational models have been developed. In this work, an agent-based model represents the malaria transmission, in which the defined agents are humans and mosquitoes. The interaction between these two agents is simulated in a spatial setting considering environmental and temporal variations. In particular, the parasite is modeled as a characteristic of both agents, considering mutation and recombination and their effects on drug resistance emergence. On the one hand, the model recreates the gonotrophic cycle of mosquito (Anopheles gambiae) and includes population births and deaths. It also represents the mosquito’s epidemiological states: susceptible, exposed and infected (SEI). On the other hand, the model includes a human population in which sulfadoxine-pyrimethamine is applied as an anti-malarial drug with 100% coverage. Human dynamics also includes births, deaths and migration. Additionally, the represented human epidemiological states in the model are SEIS, in which the reinfection probability depends on the disease exposure history. The interaction between both mosquitoes and humans occurs in a simulated scenario of one square kilometer including environmental conditions that affect the mosquito life cycle. The parasite life cycle is modeled during the interaction between mosquitoes and humans. Due to selection pressure as a consequence of sulfadoxine-pyrimethamine use, strains with fixed resistant mutations at the DHFR and DHPS loci emerge. This is modeled through the emergence of fifteen drug-resistant mutations and the subsequent recombination among alleles, generating mutational pathways developing into more resistant strains. Moreover, super-infection in humans (two genotypes) and mosquitoes (two genotypes - infection and the result of recombination) is represented. Simulations for high transmission parameters are consistent with literature, presenting coherent times for each strain appearance, and showing a mutational pathway under high transmission settings and 100% of drug coverage. Thus, measuring the prevalence of each genotype in the model allows the analysis of different cases to eradicate the disease by teh correct usage of control strategies. Furthermore, the model is being adapted for representing low transmission scenarios.

Heterogeneous feeding patterns of the dengue vector, Aedes aegypti, on individual human hosts in rural Thailand.

BackgroundMosquito biting frequency and how bites are distributed among different people can have significant epidemiologic effects. An improved understanding of mosquito vector-human interactions would refine knowledge of the entomological processes supporting pathogen transmission and could reveal targets for minimizing risk and breaking pathogen transmission cycles.Methodology and principal findingsWe used human DNA blood meal profiling of the dengue virus (DENV) vector, Aedes aegypti, to quantify its contact with human hosts and to infer epidemiologic implications of its blood feeding behavior. We determined the number of different people bitten, biting frequency by host age, size, mosquito age, and the number of times each person was bitten. Of 3,677 engorged mosquitoes collected and 1,186 complete DNA profiles, only 420 meals matched people from the study area, indicating that Ae. aegypti feed on people moving transiently through communities to conduct daily business. 10–13% of engorged mosquitoes fed on more than one person. No biting rate differences were detected between high- and low-dengue transmission seasons. We estimate that 43–46% of engorged mosquitoes bit more than one person within each gonotrophic cycle. Most multiple meals were from residents of the mosquito collection house or neighbors. People ≤25 years old were bitten less often than older people. Some hosts were fed on frequently, with three hosts bitten nine times. Interaction networks for mosquitoes and humans revealed biologically significant blood feeding hotspots, including community marketplaces.Conclusion and significanceHigh multiple-feeding rates and feeding on community visitors are likely important features in the efficient transmission and rapid spread of DENV. These results help explain why reducing vector populations alone is difficult for dengue prevention and support the argument for additional studies of mosquito feeding behavior, which when integrated with a greater understanding of human behavior will refine estimates of risk and strategies for dengue control.

Evidence of experimental vertical transmission of emerging novel ECSA genotype of Chikungunya Virus in Aedes aegypti.

BackgroundChikungunya virus (CHIKV) has emerged as one of the most important arboviruses of public health significance in the past decade. The virus is mainly maintained through human-mosquito-human cycle. Other routes of transmission and the mechanism of maintenance of the virus in nature are not clearly known. Vertical transmission may be a mechanism of sustaining the virus during inter-epidemic periods. Laboratory experiments were conducted to determine whether Aedes aegypti, a principal vector, is capable of vertically transmitting CHIKV or not.Methodology/Principal FindingsFemale Ae. aegypti were orally infected with a novel ECSA genotype of CHIKV in the 2nd gonotrophic cycle. On day 10 post infection, a non-infectious blood meal was provided to obtain another cycle of eggs. Larvae and adults developed from the eggs obtained following both infectious and non-infectious blood meal were tested for the presence of CHIKV specific RNA through real time RT-PCR. The results revealed that the larvae and adults developed from eggs derived from the infectious blood meal (2nd gonotrophic cycle) were negative for CHIKV RNA. However, the larvae and adults developed after subsequent non-infectious blood meal (3rd gonotrophic cycle) were positive with minimum filial infection rates of 28.2 (1∶35.5) and 20.2 (1∶49.5) respectively.Conclusion/SignificanceThis study is the first to confirm experimental vertical transmission of emerging novel ECSA genotype of CHIKV in Ae. aegypti from India, indicating the possibilities of occurrence of this phenomenon in nature. This evidence may have important consequence for survival of CHIKV during adverse climatic conditions and inter-epidemic periods.

Blood feeding and Plasmodium infection alters the miRNome of Anopheles stephensi.

Blood feeding is an integral process required for physiological functions and propagation of the malaria vector Anopheles. During blood feeding, presence of the malaria parasite, Plasmodium in the blood induces several host effector molecules including microRNAs which play important roles in the development and maturation of the parasite within the mosquito. The present study was undertaken to elucidate the dynamic expression of miRNAs during gonotrophic cycle and parasite development in Anopheles stephensi. Using next generation sequencing technology, we identified 126 miRNAs of which 17 were novel miRNAs. The miRNAs were further validated by northern hybridization and cloning. Blood feeding and parasitized blood feeding in the mosquitoes revealed regulation of 13 and 16 miRNAs respectively. Expression profiling of these miRNAs revealed that significant miRNAs were down-regulated upon parasitized blood feeding with a repertoire of miRNAs showing stage specific up-regulation. Expression profiles of significantly modulated miRNAs were further validated by real time PCR. Target prediction of regulated miRNAs revealed overlapping targeting by different miRNAs. These targets included several metabolic pathways including metabolic, redox homeostasis and protein processing machinery components. Our analysis revealed tight regulation of specific miRNAs post blood feeding and parasite infection in An. stephensi. Such regulated expression suggests possible role of these miRNAs during gonotrophic cycle in mosquito. Another set of miRNAs were also significantly regulated at 42 h and 5 days post infection indicating parasite stage-specific role of host miRNAs. This study will result in better understanding of the role of miRNAs during gonotrophic cycle and parasite development in mosquito and can probably facilitate in devising novel malaria control strategies at vector level.

The effects of forced-egg retention on the blood-feeding behavior and reproductive potential of Culex pipiens (Diptera: Culicidae)

High rates of West Nile virus (WNV) transmission to humans are associated with exceptionally hot and dry summers. This is paradoxical since the eggs of Culex vectors of WNV depend on the persistence of containers with water, which decline during droughts. We examined the effects of forced-egg retention on the reproductive success of female Culex pipiens as well as behavioral responses, such as likelihood of secondary blood meals. As controls we examined the effects of female age and delayed mating. We found that early mating is essential to achieve reproductive success and, consistent with an “all-or-none” ovipositing strategy, C. pipiens females are able to retain considerable reproductive potential while searching for oviposition sites. Specifically, although forced-egg retention resulted in significant decreases in fitness, the decline was moderate for 5weeks and most can be accounted for by increases in female age. Consequently, no females took blood more than once per gonotrophic cycle, which eliminates the possibility that heightened vectorial capacity due to multiple blood-feedings increases WNV transmission during periods of drought. Instead, our findings suggest that during droughts populations of C. pipiens have time to locate the remaining water holes, which are associated with human populations and WNV-competent bird species.

Alterations in mosquito behaviour by malaria parasites: potential impact on force of infection.

BackgroundA variety of studies have reported that malaria parasites alter the behaviour of mosquitoes. These behavioural alterations likely increase transmission because they reduce the risk of vector death during parasite development and increase biting after parasites become infectious.MethodsA mathematical model is used to investigate the potential impact of these behavioural alterations on the lifetime number of infectious bites delivered. The model is used to explore the importance of assumptions about the magnitude and distribution of mortality as well as the importance of extrinsic incubation period and gonotrophic cycle length. Additionally, the model is applied to four datasets taken from actual transmission settings.ResultsThe impact of behavioural changes on the relative number of lifetime bites is highly dependent on assumptions about the distribution of mortality over the mosquito-feeding cycle. Even using fairly conservative estimates of these parameters and field collected data, the model outputs suggest that altered feeding could easily cause a doubling in the force of infection.ConclusionsInfection-induced behavioural alterations have their greatest impact on the lifetime number of infectious bites in environments with high feeding-related adult mortality and many pre-infectious feeding cycles. Interventions that increase feeding-associated mortality are predicted to amplify the relative fitness benefits and hence enhance the strength of selection for behavioural alteration.