Attraction of the Mosquitoes Aedes aegypti and Aedes albopictus (Diptera: Culicidae) to a 3-Part Phytochemical Blend in a Mesocosm.

Aedes aegypti (L.) and Aedes albopictus (Skuse) mosquitoes of both sexes were attracted to a 3-part volatile synthetic phytochemical blend but differed according to their component ratios, 7:3:2 or 1:1:1, and their initial concentrations. These arbovirus vectors were presented with the blends as baits in paired baited and blank CFG traps in a large greenhouse mesocosm. Ae. aegypti attraction was highest at a 7:3:2 blend ratio, but at a concentration half that found most effective for an anopheline mosquito species in outdoor screenhouses. Both lower and higher concentrations yielded substantially lower attraction scores for Ae. aegypti. By contrast, the few tests conducted on Ae. albopictus showed that it was not as sensitive to concentration, but again it was more responsive to the 7:3:2 ratio of components than to the 1:1:1 ratio. The two sexes of both species were represented equally in the trap catches, indicating the potential value of this and similar attractive blends for population surveillance and control of Aedes mosquitoes.

Is Anopheles gambiae attraction to floral and human skin-based odours and their combination modulated by previous blood meal experience?

BACKGROUND: Mosquitoes use odours to find energy resources, blood hosts and oviposition sites. While these odour sources are normally spatio-temporally segregated in a mosquito's life history, here this study explored to what extent a combination of flower- and human-mimicking synthetic volatiles would attract the malaria vector Anopheles gambiae sensu stricto (s.s.) METHODS: In the laboratory and in large (80 m2) outdoor cages in Tanzania, nulliparous and parous A. gambiae s.s. were offered choices between a blend of human skin volatiles (Skin Lure), a blend of floral volatiles (Vectrax), or a combination thereof. The blends consisted of odours that induce distinct, non-overlapping activation patterns in the olfactory circuitry, in sensory neurons expressing olfactory receptors (ORs) and ionotropic receptors (IRs), respectively. Catches were compared between treatments. RESULTS: In the laboratory nulliparous and parous mosquitoes preferred skin odours and combinations thereof over floral odours. However, in semi-field settings nulliparous were significantly more caught with floral odours, whereas no differences were observed for parous females. Combining floral and human volatiles did not augment attractiveness. CONCLUSIONS: Nulliparous and parous A. gambiae s.s. are attracted to combinations of odours derived from spatio-temporally segregated resources in mosquito life-history (floral and human volatiles). This is favourable as mosquito populations are comprised of individuals whose nutritional and developmental state steer them to diverging odours sources, baits that attract irrespective of mosquito status could enhance overall effectiveness and use in monitoring and control. However, combinations of floral and skin odours did not augment attraction in semi-field settings, in spite of the fact that these blends activate distinct sets of sensory neurons. Instead, mosquito preference appeared to be modulated by blood meal experience from floral to a more generic attraction to odour blends. Results are discussed both from an odour coding, as well as from an application perspective.

Alteration of plant species assemblages can decrease the transmission potential of malaria mosquitoes.

Knowledge of the link between a vector population's pathogen-transmission potential and its biotic environment can generate more realistic forecasts of disease risk due to environmental change. It also can promote more effective vector control by both conventional and novel means.This study assessed the effect of particular plant species assemblages differing in nectar production on components of the vectorial capacity of the mosquito Anopheles gambiae s.s., an important vector of African malaria.We followed cohorts of mosquitoes for three weeks in greenhouse mesocosms holding nectar-poor and nectar-rich plant species by tracking daily mortalities and estimating daily biting rates and fecundities. At death, a mosquito's insemination status and wing length were determined. These life history traits allowed incorporation of larval dynamics into a vectorial capacity estimate. This new study provided both novel assemblages of putative host plants and a human blood host within a nocturnal period of maximum biting.Survivorship was significantly greater in nectar-rich environments than nectar-poor ones, resulting in greater total fecundity. Daily biting rate and fecundity per female between treatments was not detected. These results translated to greater estimated vectorial capacities in the nectar-rich environment in all four replicates of the experiment (means: 1,089.5 ± 125.2 vs. 518.3 ± 60.6). When mosquito density was made a function of survival and fecundity, rather than held constant, the difference between plant treatments was more pronounced, but so was the variance, so differences were not statistically significant. In the nectar-poor environment, females' survival suffered severely when a blood host was not provided. A sugar-accessibility experiment confirmed that Parthenium hysterophorus is a nectar-poor plant for these mosquitoes.Synthesis and applications. This study, assessing the effect of particular plant species assemblages on the vectorial capacity of malaria mosquitoes, highlights the likelihood that changes in plant communities (e.g. due to introduction of exotic or nectar-rich species) can increase malaria transmission and that a reduction of favourable nectar sources can reduce it. Also, plant communities' data can be used to identify potential high risk areas. Further studies are warranted to explore how and when management of plant species assemblages should be considered as an option in an integrated vector management strategy.

Would the control of invasive alien plants reduce malaria transmission? A review.

Vector control has been the most effective preventive measure against malaria and other vector-borne diseases. However, due to concerns such as insecticide resistance and budget shortfalls, an integrated control approach will be required to ensure sustainable, long-term effectiveness. An integrated management strategy should entail some aspects of environmental management, relying on coordination between various scientific disciplines. Here, we review one such environmental control tactic: invasive alien plant management. This covers salient plant-mosquito interactions for both terrestrial and aquatic invasive plants and how these affect a vector's ability to transmit malaria. Invasive plants tend to have longer flowering durations, more vigorous growth, and their spread can result in an increase in biomass, particularly in areas where previously little vegetation existed. Some invasive alien plants provide shelter or resting sites for adult mosquitoes and are also attractive nectar-producing hosts, enhancing their vectorial capacity. We conclude that these plants may increase malaria transmission rates in certain environments, though many questions still need to be answered, to determine how often this conclusion holds. However, in the case of aquatic invasive plants, available evidence suggests that the management of these plants would contribute to malaria control. We also examine and review the opportunities for large-scale invasive alien plant management, including options for biological control. Finally, we highlight the research priorities that must be addressed in order to ensure that integrated vector and invasive alien plant management operate in a synergistic fashion.

The Nonartemisinin Sesquiterpene Lactones Parthenin and Parthenolide Block Plasmodium falciparum Sexual Stage Transmission.

Parthenin and parthenolide are natural products that are closely related in structure to artemisinin, which is also a sesquiterpene lactone (SQL) and one of the most important antimalarial drugs available. Parthenin, like artemisinin, has an effect onPlasmodiumblood stage development. We extended the evaluation of parthenin as a potential therapeutic for the transmissible stages ofPlasmodium falciparumas it transitions between human and mosquito, with the aim of gaining potential mechanistic insight into the inhibitory activity of this compound. We posited that if parthenin targets different biological pathways in the parasite, this in turn could pave the way for the development of druggable compounds that could prevent the spread of artemisinin-resistant parasites. We examined parthenin's effect on male gamete activation and the ookinete-to-oocyst transition in the mosquito as well as on stage V gametocytes that are present in peripheral blood. Parthenin arrested parasite development for each of the stages tested. The broad inhibitory properties of parthenin on the evaluated parasite stages may suggest different mechanisms of action between parthenin and artemisinin. Parthenin's cytotoxicity notwithstanding, its demonstrated activity in this study suggests that structurally related SQLs with a better safety profile deserve further exploration. We used our battery of assays to test parthenolide, which has a more compelling safety profile. Parthenolide demonstrated activity nearly identical to that of parthenin againstP. falciparum, highlighting its potential as a possible transmission-blocking drug scaffold. We discuss the context of the evidence with respect to the next steps toward expanding the current antimalarial arsenal.

Orientation of Anopheles gambiae (Diptera: Culicidae) to Plant-Host Volatiles in a Novel Diffusion-Cage Olfactometer.

A novel diffusion-cage olfactometer tested the responses of Anopheles gambiae Giles to plant volatiles. Green-leaf volatiles are often released from cut or injured plant tissue and may alter the headspace of plants used in olfactometer assays. The diffusion-cage olfactometer is designed for use with whole, intact plants, hence giving a more realistic behavioral assay. Its simple plastic construction, ease of assembly, and accommodation to whole plants makes it a useful tool for measuring mosquito orientation to plant volatiles within large enclosures. We compared its performance to that of the more commonly used T-tube wind-tunnel olfactometer, by testing the orientation of mosquitoes to volatiles of a few prevalent plants of eastern Africa reportedly utilized by An. gambiae for sugar: Parthenium hysterophorus (Asteraceae), Ricinus communis (Euphorbiaceae), Lantana camara (Verbenaceae), and Senna occidentalis (Fabaceae). Results indicate that the diffusion-cage olfactometer is an effective alternative to conventional wind-tunnel olfactometers, to test mosquito orientation to plant volatiles under seminatural conditions.

The Invasive American Weed Parthenium hysterophorus Can Negatively Impact Malaria Control in Africa.

The direct negative effects of invasive plant species on agriculture and biodiversity are well known, but their indirect effects on human health, and particularly their interactions with disease-transmitting vectors, remains poorly explored. This study sought to investigate the impact of the invasive Neotropical weed Parthenium hysterophorus and its toxins on the survival and energy reserves of the malaria vector Anopheles gambiae. In this study, we compared the fitness of An. gambiae fed on three differentially attractive mosquito host plants and their major toxins; the highly aggressive invasive Neotropical weed Parthenium hysterophorus (Asteraceae) in East Africa and two other adapted weeds, Ricinus communis (Euphorbiaceae) and Bidens pilosa (Asteraceae). Our results showed that female An. gambiae fitness varied with host plants as females survived better and accumulated substantial energy reserves when fed on P. hysterophorus and R. communis compared to B. pilosa. Females tolerated parthenin and 1-phenylhepta-1, 3, 5-triyne, the toxins produced by P. hysterophorus and B. pilosa, respectively, but not ricinine produced by R. communis. Given that invasive plants like P. hysterophorus can suppress or even replace less competitive species that might be less suitable host-plants for arthropod disease vectors, the spread of invasive plants could lead to higher disease transmission. Parthenium hysterophorus represents a possible indirect effect of invasive plants on human health, which underpins the need to include an additional health dimension in risk-analysis modelling for invasive plants.

Evaluation of Bacillus thuringiensis israelensis as a Control Agent for Adult Anopheles gambiae.

Unlike the application of Bacillus thuringiensis israelensis (Bti) for the control of larval mosquitoes, studies of its effects on adults, for its possible use in attractive toxic sugar baits, have resulted in conflicting results. Five species have shown a decrease in adult survival due to Bti ingestion, whereas adults of Anopheles arabiensis have not. We sought to determine if ingestion of Bti by adults of Anopheles gambiae, a sibling species of An. arabiensis, increases their mortality. Laboratory-reared adults were provided continuously from emergence with water only, a sucrose solution, or a Bti suspension in sucrose solution. After 3 days, the Bti suspension was replaced with untainted sucrose solution. The mosquitoes with only water were all dead by day 3. The survivorships of those in the sucrose and sucrose-Bti treatments were insignificantly different, both with an LT50 (Lethal Time, time until 50% of individuals died) of 25 days. The results support the conclusion that adult survivorship of An. gambiae-complex members is unaffected by the ingestion of Bti in sugar meals.

Olfactory basis of floral preference of the malaria vector Anopheles gambiae (Diptera: Culicidae) among common African plants.

Mosquitoes of both sexes feed on plants to obtain sugar. Nocturnal species probably locate the plants primarily by their volatile semiochemicals that also form the basis for the mosquitoes' innate plant-species preferences. To evaluate these olfactory preferences quantitatively, we used a two-choice wind-tunnel olfactometer to measure the upwind orientation of Anopheles gambiae Giles, an important vector of malaria in equatorial Africa, toward odor plumes produced by nine plant species common where this mosquito occurs. These plants are reported to induce feeding behaviors in An. gambiae and to produce floral or extrafloral nectar. Results presented here demonstrated that the volatiles of S. didymobotrya, P. hysterophorus, S. occidentalis, and L. camara, in descending order of numbers of mosquitoes responding, were all attractive, compared to a control plant species, whereas D. stramonium, R. communis, S. bicapsularis, T. stans, and T. diversifolia were not. As expected, chromatographic analysis of the headspace of attractive plants whose volatiles were captured by stir-bar sorptive extraction revealed a wide range of compounds, primarily terpenoids. Once their bioactivity and attractiveness for An. gambiae, alone and in blends, has been firmly established, some of these semiochemicals may have applications in population sampling and control.

Delayed egg hatching of Anopheles gambiae (Diptera: Culicidae) pending water agitation.

Mosquito eggs laid on water surfaces typically hatch spontaneously soon after the embryos within them become fully formed first-instar larvae. However, we have found that Anopheles gambiae Giles, an important vector of malaria in Africa, exhibits delayed hatching until the water surface is agitated, a feature overlooked in most laboratory colonies. Agitation within 24 h postoviposition, before embryonation was complete, failed to stimulate delayed postembryonic hatching of isolated eggs on the following day (day 2), when < 1% had hatched spontaneously. However, 5 min of water agitation of these dormant pharate first-instar larvae on day 2 resulted in an almost immediate hatch of 63.3 versus 0% of nonagitated controls, plus another 3.9 versus 0.3%, respectively, during the following 24 h. With daily agitation, installment hatching occurred mainly during 2-6 d postoviposition. The mean cumulative hatch after 7 d of daily agitation was 83.1 versus 1.1% of nonagitated eggs. Experiments with eggs in groups demonstrated that egg density and activity of already-hatched larvae had no stimulatory effect. Eggs stored 1-4 wk at 25.5 or at 15.5 degrees C, and then agitated daily for 6 d at 25.5 degrees C, showed a gradual decline in viability. Viability was sustained longer at the lower temperature. Implications of agitation-induced egg hatching for rainy-season and dry-season ecology of An. gambiae are discussed. Suspended hatching and cool storage already are proving convenient for efficient mass rearing and accurate modeling of weather-based population dynamics.

Development and assessment of plant-based synthetic odor baits for surveillance and control of malaria vectors.

BACKGROUND: Recent malaria vector control measures have considerably reduced indoor biting mosquito populations. However, reducing the outdoor biting populations remains a challenge because of the unavailability of appropriate lures to achieve this. This study sought to test the efficacy of plant-based synthetic odor baits in trapping outdoor populations of malaria vectors. METHODOLOGY AND PRINCIPAL FINDING: Three plant-based lures ((E)-linalool oxide [LO], (E)-linalool oxide and (E)-ß-ocimene [LO + OC], and a six-component blend comprising (E)-linalool oxide, (E)-ß-ocimene, hexanal, ß-pinene, limonene, and (E)-ß-farnesene [Blend C]), were tested alongside an animal/human-based synthetic lure (comprising heptanal, octanal, nonanal, and decanal [Blend F]) and worn socks in a malaria endemic zone in the western part of Kenya. Mosquito Magnet-X (MM-X) and lightless Centre for Disease Control (CDC) light traps were used. Odor-baited traps were compared with traps baited with either solvent alone or solvent + carbon dioxide (controls) for 18 days in a series of randomized incomplete-block designs of days × sites × treatments. The interactive effect of plant and animal/human odor was also tested by combining LO with either Blend F or worn socks. Our results show that irrespective of trap type, traps baited with synthetic plant odors compared favorably to the same traps baited with synthetic animal odors and worn socks in trapping malaria vectors, relative to the controls. Combining LO and worn socks enhanced trap captures of Anopheles species while LO + Blend F recorded reduced trap capture. Carbon dioxide enhanced total trap capture of both plant- and animal/human-derived odors. However, significantly higher proportions of male and engorged female Anopheles gambiae s.l. were caught when the odor treatments did not include carbon dioxide. CONCLUSION AND SIGNIFICANCE: The results highlight the potential of plant-based odors and specifically linalool oxide, with or without carbon dioxide, for surveillance and mass trapping of malaria vectors.

Scanning electron microscopy of eggs of Sabethes cyaneus.

Mosquitoes of the Neotropical genus Sabethes, some species of which are yellow fever vectors, most often develop through the immature stages in tree holes. Sabethes eggs have not been previously characterized using scanning electron microscopy. Eggs of Sabethes cyaneus (length: 349.6 +/- 2.7 microm; width: 172.6 +/- 1.14 microm; n = 10) are almost biconical when examined from the top. From a lateral perspective 2 surfaces can be seen. One surface is smooth and more convex, whereas the other is less convex and partially covered by a network from which many fungiform tubercles arise. The micropyle is situated on the smooth surface of the pointed anterior tip and is surrounded by an irregular row of tubercles, some of which are leaf shaped. No structures possibly involved in adhesion to surfaces are visible. When hatching, the egg splits dorsoventrally approximately two-thirds of the length from the anterior end. The tubercles appear to be water repellent, and the more convex/smoother surface is downturned, and this position on water was confirmed by direct observation. The eggs float free on the water surface.

Rapid discrimination between Anopheles gambiae s.s. and Anopheles arabiensis by High-Resolution Melt (HRM) analysis.

There is a need for more cost-effective options to more accurately discriminate among members of the Anopheles gambiae complex, particularly An. gambiae and Anopheles arabiensis. These species are morphologically indistinguishable in the adult stage, have overlapping distributions, but are behaviorally and ecologically different, yet both are efficient vectors of malaria in equatorial Africa. The method described here, High-Resolution Melt (HRM) analysis, takes advantage of minute differences in DNA melting characteristics, depending on the number of incongruent single nucleotide polymorphisms in an intragenic spacer region of the X-chromosome-based ribosomal DNA. The two species in question differ by an average of 13 single-nucleotide polymorphisms giving widely divergent melting curves. A real-time PCR system, Bio-Rad CFX96, was used in combination with a dsDNA-specific dye, EvaGreen, to detect and measure the melting properties of the amplicon generated from leg-extracted DNA of selected mosquitoes. Results with seven individuals from pure colonies of known species, as well as 10 field-captured individuals unambiguously identified by DNA sequencing, demonstrated that the method provided a high level of accuracy. The method was used to identify 86 field mosquitoes through the assignment of each to the two common clusters with a high degree of certainty. Each cluster was defined by individuals from pure colonies. HRM analysis is simpler to use than most other methods and provides comparable or more accurate discrimination between the two sibling species but requires a specialized melt-analysis instrument and software.

Effects of plant-community composition on the vectorial capacity and fitness of the malaria mosquito Anopheles gambiae.

Dynamics of Anopheles gambiae abundance and malaria transmission potential rely strongly on environmental conditions. Female and male An. gambiae use sugar and are affected by its absence, but how the presence or absence of nectariferous plants affects An. gambiae abundance and vectorial capacity has not been studied. We report on four replicates of a cohort study performed in mesocosms with sugar-poor and sugar-rich plants, in which we measured mosquito survival, biting rates, and fecundity. Survivorship was greater with access to sugar-rich plant species, and mortality patterns were age-dependent. Sugar-poor populations experienced Weibull mortality patterns, and of four populations in the sugar-rich environment, two female and three male subpopulations were better fitted by Gompertz functions. A tendency toward higher biting rates in sugar-poor mesocosms, particularly for young females, was found. Therefore, vectorial capacity was pulled in opposing directions by nectar availability, resulting in highly variable vectorial capacity values.

Analysis and optimization of a synthetic milkweed floral attractant for mosquitoes.

A pentane extract of flowers of common milkweed, Asclepias syriaca (Asclepiadaceae), elicited significant orientation from both male and female Culex pipiens in a dual-port flight olfactometer. Analysis of the extract by gas chromatography-mass spectrometry revealed six major constituents in order of relative abundance: benzaldehyde, (E)-ß-ocimene, phenylacetaldehyde, benzyl alcohol, nonanal, and (E)-2-nonenal. Although not all were collected from the headspace profile of live flowers, a synthetic blend of these six compounds, when presented to mosquitoes in the same levels and proportions that occur in the extract, elicited a response comparable to the extract. Subtractive behavioral bioassays demonstrated that a three-component blend consisting of benzaldehyde, phenylacetaldehyde, and (E)-2-nonenal was as attractive as the full blend. These findings suggest the potential use of synthetic floral-odor blends for monitoring or control of both male and female disease-vectoring mosquitoes.

Effects of bed net use, female size, and plant abundance on the first meal choice (blood vs sugar) of the malaria mosquito Anopheles gambiae.

BACKGROUND: The purpose of this study was to determine whether the sugar-or-blood meal choice of Anopheles gambiae females one day after emergence is influenced by blood-host presence and accessibility, nectariferous plant abundance, and female size. This tested the hypothesis that the initial meal of female An. gambiae is sugar, even when a blood host is available throughout the night, and, if not, whether the use of a bed net diverts mosquitoes to sugar sources. METHODS: Females and males <1-day post-emergence were released in a mesocosm. Overnight they had access to either one or six Senna didymobotrya plants. Simultaneously they had access to a human blood host, either for 8 h or for only 30 min at dusk and dawn (the remainder of the night being excluded by an untreated bed net). In a third situation, the blood host was not present. All mosquitoes were collected in the morning. Their wing lengths, an indicator of pre-meal energetic state, were measured, and their meal choice was determined by the presence of midgut blood and of fructose. RESULTS: Female sugar feeding after emergence was facultative. When a blood host was accessible for 8 h per night, 92% contained blood, and only 3.7% contained sugar. Even with the use of a bed net, 78% managed to obtain a blood meal during the 30 min of accessibility at dusk or dawn, but 14% of females were now fructose-positive. In the absence of a blood host, and when either one or six plants were available, a total of 21.7% and 23.6% of females and 30.8% and 43.5% of males contained fructose, respectively. Feeding on both sugar and blood was more likely with bed net use and with greater plant abundance. Further, mosquitoes that fed on both resources were more often small and had taken a sugar meal earlier than the blood meal. The abundance of sugar hosts also affected the probability of sugar feeding by males and the amount of fructose obtained by both males and females. CONCLUSION: Even in an abundance of potential sugar sources, female An. gambiae appear to prefer a nearby human source of blood. However, the decision to take sugar was more likely if energy reserves were low. Results probably would differ if sugar hosts were more attractive or yielded larger sugar meals. The diversion of energetically deprived mosquitoes to sugar sources suggests a possible synergy between bed nets and sugar-based control methods.

Effect of sugar on male Anopheles gambiae mating performance, as modified by temperature, space, and body size.

BACKGROUND: Anopheles gambiae plant-sugar feeding was thought to be rare and physiologically optional. Unlike adult females, males have no alternative source of energy and soon die with only water, yet they might be competent to inseminate all females within their brief lifespan. This study was designed to detect sugar's effect, if any, on male performance. METHODS: Males with and without 20% sucrose were evaluated at two body sizes and two temperatures, 23 degrees and 27 degrees C. Survival was recorded twice daily, and sexual behaviour was recorded each night after adult emergence. Insemination at a 2:1 male:female ratio was examined in three cage sizes, including walk-in mesocosms. RESULTS: Without sugar, males of both sizes lived longer at 23 degrees than 27 degrees C, and large males lived longer at each temperature. Survival of large males at low temperature averaged 3.7 days, small males at high temperature, 1.9 days. With sugar, males in all four treatments suffered minimal mortality. With sugar, in small cages, large males at 27 degrees C matured most rapidly. A few erected fibrillae and inseminated females on night 1. On night 2, maximal proportions erected fibrillae and swarmed, and over one-third of females became inseminated. Small sugar-fed males at 23 degrees C matured most slowly but had achieved nearly maximal levels of swarming by night 3. By night 5, small males had inseminated more than half the females, and large males had inseminated nearly all of them. Without sugar, large males progressed similarly during the first two nights. On night 3, however, the proportion erecting fibrillae and swarming declined precipitously at 27 degrees C, and to a lesser degree at 23 degrees C. Cumulative insemination never reached high levels. Small males never achieved high levels of fibrillar erection or swarming and inseminated few females, even at 23 degrees C. In larger cages and under more semi-natural conditions, regardless of body size, without sugar male insemination capacity was virtually nonexistent. CONCLUSION: Under some conditions, a limited number of sugar-deprived males can survive long enough to inseminate females. However, in nature males that cannot obtain sugar at frequent intervals will not be competitive with those that can, suggesting that male performance is closely tied to plant communities.

An effective indoor mesocosm for studying populations of Anopheles gambiae in temperate climates.

To study the indoor behavior of Anopheles gambiae populations in a temperate climate, we have devised a walk-in mesocosm, built within a greenhouse. The structure provides conditions more natural than laboratory cages, including sufficient room for swarming and for flight between resting sites, sugar-bearing plants, a human host, and an oviposition site. These activities impose energy demands closer to those encountered in the field. The structure also has predators, fluctuating temperatures, natural daylight, and an evening crepuscular period. Most important, its resting sites comprise a bank of tubes that can be inspected or removed individually to obtain, at regular time intervals, random representative samples of an experimental population while all individuals are inactive. Samples from aging cohorts of mosquitoes, released at emergence, can yield information on behavioral sequences, mate competition, reproductive success, and survival under different nutritional regimes.

Internalization of LDL-receptor superfamily yolk-protein receptors during mosquito oogenesis involves transcriptional regulation of PTB-domain adaptors.

In the anautogenous disease vector mosquitoes Anopheles gambiae and Aedes aegypti, egg development is nutritionally controlled. A blood meal permits further maturation of developmentally repressed previtellogenic egg chambers. This entails massive storage of extraovarian yolk precursors by the oocyte, which occurs through a burst of clathrin-mediated endocytosis. Yolk precursors are concentrated at clathrin-coated structures on the oolemma by two endocytic receptors, the vitellogenin and lipophorin receptors. Both these mosquito receptors are members of the low-density-lipoprotein-receptor superfamily that contain FxNPxY-type internalization signals. In mammals, this tyrosine-based signal is not decoded by the endocytic AP-2 adaptor complex directly. Instead, two functionally redundant phosphotyrosine-binding domain adaptors, Disabled 2 and the autosomal recessive hypercholesterolemia protein (ARH) manage the internalization of the FxNPxY sorting signal. Here, we report that a mosquito ARH-like protein, which we designate trephin, possess similar functional properties to the orthologous vertebrate proteins despite engaging AP-2 in an atypical manner, and that mRNA expression in the egg chamber is strongly upregulated shortly following a blood meal. Temporally regulated trephin transcription and translation suggests a mechanism for controlling yolk uptake when vitellogenin and lipophorin receptors are expressed and clathrin coats operate in previtellogenic ovaries.

Phytochemicals as population sampling lures.

Most mosquito species have 2 hosts: vertebrate animals and vascular plants. The kairomones of vertebrates have been employed extensively as attractants in traps that are used for surveillance, either to assess adult density of mosquito populations or to detect pathogen activity. They also have been employed in basic field studies of mosquito physiology, behavior, and ecology. The semiochemicals that mosquitoes use to find plant hosts for their sugar, by contrast, have not been utilized at all. Currently we are characterizing attractive blends of volatile compounds produced by plant species visited by Aedes vexans, Culex pipiens, and Anopheles gambiae. These blends may be effective in attracting a unique subset of a mosquito population when deployed in surveillance traps. The principal advantages of phytochemical attractants are that they lure a) both sexes, b) all ages, including those that are newly emerged, c) females in all gonotrophic states, and d) both nondiapausing and reproductively diapausing females. Potential challenges to their successful use are the abundance of competing volatiles, narrow plant-host specificity, and a weaker behavioral response to phytochemical cues.