Aedes aegypti: Sensilla trichodea and stimulus-conducting structures

Much of the morphology of the olfactory sensilla on the antennae of the mosquito Aedes aegypti (L.) has been described, however little is known about the fate of odour molecules once they have been adsorbed onto the surfaces of sensilla. A stimulus-conducting system of pores, pore kettles, and pore tubules has been described for the sensilla trichodea (olfactory hairs) of several insects but not mosquitoes. Scanning electron microscopy was used to identify the s. trichodea of Ae. aegypti and to attempt visualization of their pore openings. Chemical fixation, cryopreparation, freeze drying, and negative staining, with high resolution transmission electron microscopy (TEM), were used to locate putative stimulus-conducting structures associated with the pores. TEM sections using Dalton's fixative or freeze drying showed pores and pore tubules, whereas pore tubules were poorly preserved in cryoprepared sections. The putative stimulus-conducting structures were clearly demonstrated by negative staining of whole sensilla which was quick and easy. The current hypothesis of olfactory stimulus conduction is extended to include Ae. aegypti.     

Flight behavior and phase polymorphism in the mosquito Aedes taeniorhynchus

Adults of Aedes taeniorhynchus, reared under two different population densities at 27° under LD 12:12, exhibit differences in their flight behavior. Adults reared under crowded conditions show a main flight peak on the day of emergence as compared to adults reared under non-crowded conditions, which do not. On the basis of their flight activity, these adults are designated as belonging to migrant and non-migrant phases. Adults from both these conditions did not show statistically significant differences in their morphological characteristics, percent glycogen and state of autogeny, but those belonging to migrant phase are much lighter in dry body weight and have lower percentages of lipid than those belonging to non-migrant phase. Analysis of these results and earlier work on this insect suggests that phase polymorphism in this insect is due to the plasticity in its ontogenetic potential.

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Fluggewohnheiten und phasenpolymorphismus bei der mücke Aedes taeniorhynchus

Zusammenfassung Erwachsene Aedes taeniorhynchus, die unter 27° und LD 12:12 bei zwei unterschiedlichen Populationsdichten aufgezogen wurden, weisen Unterschiede in ihrem Flugverhalten auf. Unter Gedrängebedingungen aufgewachsene Imagines zeigen am Schlupftage einen Hauptfluggipfel, nicht gedrängt aufgezogene dagegen nicht. Auf Grund ihrer Flugaktivität werden diese Adulten als Angehörige der migranten und der non-migranten Phase bezeichnet. Die unter diesen verschiedenen Bedingungen aufgezogenen Imagines weisen in ihren morphologischen Eigenschaften, dem prozentualen Glykogengehalt und im Autogeniezustand keine statistisch signifikanten Unterschiede auf, aber die zur Migrantenphase gehörenden haben ein viel geringeres Trockengewicht und geringere Prozentgehalte an Lipiden als die zur nonmigranten Phase zu rechnenden. Die Analyse dieser Ergebnisse und frühere Untersuchungen lassen vermuten, daß der Phasenpolymorphismus bei diesem Insekt eine Folge der Plastizität seines ontogenetischen Potentials ist.

     

Identification of novel LTR retrotransposons in the genome of Aedes aegypti

We have detected seventy-six novel LTR retrotransposons in the genome of the mosquito Aedes aegypti by a genome wide analysis using the LTR_STRUC program. We have performed a phylogenetic classification of these novel elements and a distribution analysis in the genome of A. aegypti. These mobile elements belong either to the Ty3/gypsy or to the Bel family of retrotransposons and were not annotated in the mosquito LTR retrotransposon database (TEfam). We have found that  1.8% of the genome is occupied by these newly detected retrotransposons that are distributed predominantly in intergenic genomic sequences and introns. The potential role of retrotransposon insertions linked to host genes is described and discussed. We show that a retrotransposon family belonging to the Osvaldo lineage has peculiar structural features, and its presence is likely to be restricted to the A. aegypti and to the Culex pipiens quinquefasciatus genomes. Furthermore we show that the ninja-like group of elements lacks the Primer Binding Site (PBS) sequence necessary for the replication of retrotransposons. These results integrate the knowledge on the complicate genomic structure of an important disease vector.     

Sampling a population of Aedes taeniorhynchus in a large outdoor cage

A comparison was made in a 16-sided outdoor cage measuring 15 m in diameter of the numbers and physiological stages of mosquitoes taken in a light trap sample from a known population. For each brood, six suction and five light collections were made each night for six successive nights. Both response to light and flight activity tended to increase with age. However, neither the numbers taken at different times during the night nor the ovarian composition of the samples were comparable to collections made outdoors. These differences were probably caused by the contrast between illumination levels inside and outside the cage interacting with the circadian rhythm of crepuscular activity and the physical restrictions of the cage.

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Résumé Le but de cette étude est de déterminer le rapport existant entre un échantillonnage par piège lumineux et la population réelle.Des moustiques, venus d'atteindre le stade imaginal, sont relâchés dans une vaste cage de 15 m de diamètre et de 2.5 m d'hauteur, puis sont seulement alimentés de sucre. La proportion de femelles autogènes, le taux d'accouplements, et les réserves énergétiques de chaque couvée ont été précisés. Pour chaque couvée, un mécanisme d'échantillonnage fait alternativement des échantillons par aspiration ou par lumière (en realité aspiration et lumière) pendant six nuits successives. L'échantillon par aspiration 1^er commençait au coucher de soleil et terminait au crépuscule astronomique. Un échantillon (échantillon 2^me), de duration 5 minutes, se faisait à cette heure et encore à 2200 (échantillon 4^me), 0000 (échantillon 6^me), 0200 (échantillon 8^me) et au crépuscule astronomique du matin (échantillon 10^me). Les échantillons 3^me, 5^me, 7^me, 9^me et 11^me occupaient toutes les autres heures. Du clair de lune simulé se fournissait pour maintenir un niveau uniforme d'illumination pendant toutes les heures sauf les temps des échantillons crépusculaires. En compensation pour les durées inégales des échantillons par aspiration, on a reduit les données au nombre d'individus capturés en 5 minutes. Un échantillon à filet de balayage se faisait suivant les opérations de chaque nuit. Chaque jour on collectionnait les moustiques morts et on estimait les moustiques vivants dans la cage. On examinait les moustiques capturés pour déterminer l'état des ovaires des femelles. En addition, on opérait le mécanisme d'échantillonnage au dehors de la cage.Chaque couvée possédait ses propres caractéristiques concernant la proportion de femelles autogènes et accouplés, et pour les deux sexes la proportion alimenté de sucre, les réserves énergétiques, la mortalité, la réaction à la lumière et le niveau de l'activité du vol. Pour les femelles le niveau de l'activité du vol et la réaction à la lumière semblaient à augmenter avec l'âge, et la réaction à la lumière accroissait plus rapidement qu'accroissait l'activité du vol. A des heures différentes de la nuit il y avait peu de variation d'état des ovaires des femelles échantillonnées, mais il y avait plus de variation entre les échantillons par les méthodes différentes; par example, la proportion la plus petite des femelles gravides s'échantillonnait par filet de balayage et la proportion la plus grande par piège lumineux. La proportion la plus grande des moustiques alimentés de sucre aussi s'échantillonnait par filet de balayage.Les échantillons par aspiration 1^er et 11^me étaient petits et ne montraient pas des cimes d'activité crépusculaire, tandis que les échantillons 7^me et 9^me étaient plus grands que les 3e et 5^me. Les échantillons par piège de lumière 2e et 10^me prenaient deux fois plus des femelles que chacun des autres échantillons par piège de lumière. Le dessin des échantillons par aspiration et par piège de lumière dans la cage ne ressemblait pas au dessin au dehors. Ces différences probablement ont été causées par le contraste entre les niveaux d'illumination à l'intérieur et à l'extérieur de la cage qui s'entragit avec le rhythme circadien d'activité crépusculaire et les restrictions physiques imposées par la cage. Le comportement normal n'était pas possible et les résultats d'échantillonnage obtenus dans une cage, même tellement grande, n'étaient pas comparable aux résultats aux conditions naturelles.

     

Effects of antennectomy on blood-feeding behavior of Aedes aegypti mosquitoes

Completely flagellectomized females of Aedes aegypti usually do not locate a person of moderate attractiveness at very close range (0 to 4.5 cm from hand), but all may feed on an individual of great attractiveness under similar conditions. Approximately seven flagellar segments on one or two antennae are normally required for a high degree of blood avidity towards a person of average attractiveness in a cubic cage with 30.5-cm sides.

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Résumé Le test consiste à offrir la main et (ou) l'avant-bras d'un individu préalablement reconnu peu ou très attractif, à des moustiques enfermés dans de petites cages cylindriques plus ou moins longues; cette longueur conditionne la distance à laquelle les moustiques peuvent déceler l'appât.L'ablation bilatérale des flagelles antennaires supprime toute alimentation des moustiques opérés, sur un individu reconnu comme modérément attractif, mais ces mêmes insectes peuvent encore déceler un individu hautement attractif et s'alimenter. Les diverses amputations pratiquées montrent qu'il faut que soient conservés approximativement sept segments flagellaires (répartis sur une ou deux antennes) pour maintenir un comportement hématophage à l'égard d'une personne médiocrement attractive, les insectes étant enfermés dans une cage de 30,5×30,5×30,5 cm.

     

Effects of AeDNV infection on Aedes aegypti lifespan and reproduction

The effects of Aedes Densovirus (AeDNV) infections on survival, fertility, fecundity and vertical transmission in Aedes aegypti (Diptera: Culicidae) were measured in laboratories in Kiev, Ukraine and Colorado, USA and incorporated into a predictive model of the effects of AeDNV on vector capacity. Adult lifespan and daily survival were reduced in AeDNV infected mosquitoes. This effect was dependent on the dose of the virus. Infected females had decreased fecundity. The oviposition rate was less in infected females and the hatch rate declined in eggs laid by infected females. The amounts of AeDNV in infected females and the infection rate of their offspring were measured with real-time PCR. The average filial transmission rate was 70% and larval infection rates from infected females varied between 42 and 62%. Vertically infected larvae, and individual eggs contained 1 × 10⁵ AeDNV genome equivalents (geq). Modeling the effects of AeDNV infection on Ae. aegypti populations suggested a large decrease in the numbers of eggs, larvae, pupae, and adults arising from infected mothers and suggested that AeDNV treatment of larvae could cause up to a 76% reduction of infectious mosquito days.     

Some effects of overcrowding on the respiration of larval Aedes aegypti

Changes in the respiratory rates of standard, overcrowded, and starved fourth-instar Aedes aegypti are compared. The respiratory effects of varying larval densities during and prior to measurement are investigated. In addition, the results of exposure of standard and over-crowded larvae to growth-retarding factors (GRF) at various concentrations are also evaluated.

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Résumé Le but de cette étude est d'étudier les modifications de l'activité respiratoire chez des larves du 4ème stade mises respectivement en condition d'élevage standard, en condition de surpeuplement, et en condition de restriction alimentaire.Les conditions de surpeuplement et de restriction alimentaire entrainent un abaissement significatif du niveau du métabolisme respiratoire. L'activité respiratoire des larves élevées en condition standard, prend des valeurs plus élevées quand la mesure est faite sur un plus grand nombre de larves rassemblées dans le tube-test; par contre les conditions de mesure (15, 25 ou 35 larves dans le tube-test) ne modifient pas la valeur de l'activité respiratoire des larves élevées en condition de surpeuplement. Quand des larves élevées en milieu surpeuplé et en milieu standard sont soumises à de fortes concentrations des facteurs retardant la croissance (Growth-retardant factors; GRF de Moore & Fisher 1969) l'activité respiratoire est stimulée. Dans les conditions de surpeuplement et de restriction alimentaire, deux types facteurs pourraient agir sur le métabolisme respiratoire, des facteurs chimiques et des facteurs mécaniques tels que l'agitation des individus dans un espace étroit (par exemple le tube-test).

     

Biochemical analysis of a blood meal-induced Aedes aegypti glutamine synthetase gene

Glutamine synthetase (GS) in the mosquito, Aedes aegypti, is induced in the midgut following a blood meal. Mosquito GS message is detected as soon as 1 h post-blood feeding and remains stable for 18 h. Using a PCR product encoding mosquito GS, a λgt10 adult female mosquito cDNA library was screened. A cDNA clone, pCl5A2, encoding the full translation product of mosquito GS was isolated and sequence analyses performed. Mosquito GS cDNA is 2.5 kb in length and its putative translation product shares all the conserved regions characteristic of the GS gene family, including the presumed ATP biding site. Glutamine synthetase activity in the mosquito midgut is highest at 18 h post-blood feeding. Activity can be detected over a broad pH range, from 6.0 to 7.5. Unlike other cellular GS enzymes, mosquito GS is not active in the presence of ATP. Very low dosages (0.05 mM) of L-methionine S-sulfoximine are sufficient to partially inhibit mosquito GS activity. Inhibition of GS disrupts the normal formation of the midgut peritrophic matrix, suggesting that GS enzyme might be involved in the initial pathway of chitin synthesis. The unique expression pattern and inducible nature of the mosquito GS gene make it an interesting candidate for studying promoter function. Additionally, the blood meal activation of the GS gene makes this a potentially valuable tool in mosquito transformation studies.     

Vitelline envelope genes of the yellow fever mosquito, Aedes aegypti

Vitelline envelope genes from the mosquito Aedes aegypti were analyzed with respect to their DNA sequences, genomic representation, temporal and spatial expression profiles and response to 20-hydroxyecdysone. Genomic clones of three vitelline envelope genes, 15a-1, 15a-2 and 15a-3 were isolated. Southern analysis indicates that all three genes are represented by a single copy in the genome. The deduced amino acid sequences of all three vitelline envelope genes contain a conserved region of 46 residues that overlaps with a region that is conserved in four Drosophila melanogaster vitelline envelope genes. DNA was sequenced flanking the 15a-1, 15a-2 and 15a-3 coding regions. A 360 bp sequence 5′ of the 15a-2 coding region was identified with 72% identity to a sequence upstream of the Ae. aegypti VgA1 vitellogenin gene. The temporal patterns of 15a-1, 15a-2 and 15a-3 expression, as determined by Northern analysis, were similar. The spatial patterns of expression, as determined by whole-mount in situ hybridization, differed between the three genes. 15a-1 and 15a-3 were only expressed in the middle and posterior regions of the follicle, while 15a-2 was also expressed at the anterior region. Vitelline envelope gene expression was higher in ovaries that were dissected at 0, 2 and 10 h following a blood meal and then incubated in vitro for 10 h in medium containing 10⁻⁵ M 20-hydroxyecdysone, compared to ovaries that were incubated without hormone.     

Lipophorin as a yolk protein precursor in the mosquito, Aedes aegypti

We examined expression of the lipophorin (Lp) gene, lipophorin (Lp) synthesis and secretion in the mosquito fat body, as well as dynamic changes in levels of this lipoprotein in the hemolymph and ovaries, during the first vitellogenic cycle of females of the yellow fever mosquito, Aedes aegypti. Lipophorin was purified by potassium bromide (KBr) density gradient ultracentrifugation and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were produced against individual Lp apoproteins, apolipoprotein-I (apoLp-I) and apolipoprotein-II (apoLp-II), with molecular weights of 240 and 75 kDa, respectively. We report here that in the mosquito A. aegypti, Lp was synthesized by the fat body, with a low level of the Lp gene expression and protein synthesis being maintained in pre- and postvitellogenic females. Following a blood meal, the Lp gene expression and protein synthesis were significantly upregulated. Our findings showed that the fat body levels of Lp mRNA and the rate of Lp secretion by this tissue reached their maximum at 18 h post-blood meal (PMB). 20-Hydroxyecdysone was responsible for an increase in the Lp gene expression and Lp protein synthesis in the mosquito fat body. Finally, the immunocytochemical localization of Lp showed that in vitellogenic female mosquitoes, this protein was accumulated by developing oocytes where it was deposited in yolk granules.     

Alteration of feeding behavior and fertility in Aedes aegypti by the chemosterilant apholate

Studies have demonstrated that the inhibition of ovarian development, as contrasted to inhibition of oogenesis, by the chemosterilant Apholate, causes a significant increase in the biting rate of female Aedes aegypti. The material also reduces the fertility of males of this species.The possibility exists that materials with similar modes of action might increase the vector efficiency of this species, at least temporarily, while the population is being reduced at a slower rate. Assessment of this possibility should be made in other vectors, of both plant and animal diseases, as well as with other chemosterilants.

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Zusammenfassung Untersuchungen mit dem Chemosterilans Apholat weisen eine zweifache Wirkung auf Aedes aegypti nach. Werden frisch geschlüpfte Larven 10–20 ppm des Stoffes ausgesetzt, so werden die Prozesse der Oogenese und Spermatogenese in so starkem Maße beeinflußt, daß ein hoher Anteil unfruchtbarer Eier abgelegt wird. Bei einer Konzentration von 30 ppm wird jedoch die Entwicklung der Ovarien selbst gehemmt, so daß den Erwachsenen diese Organe fehlen. Solche Weibchen nahmen signifikant mehr Blut auf als unbehandelte Weibchen, obwohl gar keine Eier gebildet werden konnten.Diese Änderung im Nahrungsaufnahme-Verhalten könnte wichtige Folgen für die Übertragung von Pathogenen durch behandelte Vektoren von Tier- und Pflanzenkrankheiten haben. Einige Aspekte dieses Problems werden erörtert.

     

Chemical composition and larvicidal effects of essential oil of Dendropanax morbifera against Aedes aegypti L.

Essential oils obtained from the flowers of Dendropanax morbifera were extracted and the chemical composition and larvicidal effects were studied. The analyses were conducted by gas chromatography and mass spectroscopy (GC–MS) revealed that the essential oil of D. morbifera contained 27 compounds. The major chemical components identified were γ-elemene (18.59%), tetramethyltricyclohydrocarbon (10.82%), β-selinene (10.41%), α-zingibirene (10.52%), 2-isopropyl-5-methylbicylodecen (4.2%), β-cubebene (4.19), and 2,6-bis(1,1-Dimethylethyl)-4-phenol (4.01%). The essential oil had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L. with an LC₅₀ value of 62.32 ppm and an LC₉₀ value of 131.21 ppm. The results could be useful in search for newer, safer, and more effective natural larvicidal agents against A. aegypti.     

Factors affecting polybrene-mediated transfection of cultured Aedes albopictus (mosquito) cells

The polycation 1,5-dimethyl-1,5-diazaundecamethylene polymethobromide (polybrene) is superior to calcium phosphate for the introduction of purified DNA into cultured Aedes albopictus (mosquito) cells. Adsorption of the polybrene-DNA complex to mosquito cells was essentially linear for 6 h. However, the rate of adsorption of DNA increased when the DNA-polybrene mixture was preincubated for several hours prior to addition to cells. A recombinant plasmid carrying an inducible chloramphenicol acetyltransferase gene under the control of a Drosophila heat shock protein (hsp) promoter was used to show that expression of transfected DNA was highest when cells were treated with a freshly prepared polybrene-DNA mixture. Optimal expression was observed in cells transfected with 4-13 micrograms of DNA per 10(6) cells; transfection with 24 micrograms of DNA resulted in reduced CAT expression. Variation in the polybrene-DNA ratio improved transfection with high levels of DNA. In mosquito cells, CAT expression was independent of DNA methylation.     

Lecithin in synthetic larval diet for Aedes aegypti improves larval and adult performance

A chemically defined synthetic rearing medium was used to compare larval growth of Aedes aegypti with or without crude animal lecithin or synthetic dipalmitoyl lecithin. Pupal weights, adult female life spans and oviposition histories of pupae and adults derived from synthetic diet rearings and from crude culture (liver powder) rearings were also compared. Both lecithins improved larval growth rate; optimal concentrations reduced time to pupation by 2 and 1 days for animal and synthetic lecithins respectively, and animal lecithin was better tolerated by the mosquito larvae at the higher concentrations tested. Addition of crude animal lecithin to the basal synthetic rearing medium had little effect on the weights of male and female pupae, but it increased adult female life spans and improved weekly egg production. In no case did mosquitoes from axenic rearings in synthetic media deviate greatly from limits specified by earlier workers for field-derived mosquitoes.

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Résumé Les développements larvaires d'Aedes aegypti ont été comparés sur des substrats alimentaires avec ou sans lecithine animale brute ou DL-a-dipalmitoyle lecithine synthétique.Les comparaisons ont porté aussi sur les poids nymphaux, la longé-vité et la fécondité de femelles élevées sur régime synthétique ou à partir de poudre de foie.Les deux types de lecithines améliorent le développement larvaire; les concentrations optimales des deux régimes (lecithine animale et lecithine synthétique) avancent respectivement la pupaison de 2 et 1 jour; les lecithines animale et synthétique sont tolérées par les larves respectivement jusqu'aux concentrations de 0,008 et 0,006%.L'addition de lecithine animale brute au régime contenant de la lecithine synthétique a eu peu d'effets sur les poids des nymphes mâles et femelles mais a augmenté la longévité et la fécondité hebdomadaire des femelles.Les performances des moustiques élevés sur substrat aseptique synthétique n'ont jamais été très éloignées des limites indiquées antérieurement par les travaux sur moustiques provenant de la nature.

     

Sensitivity of Aedes aegypti adults (Diptera: Culicidae) to the vapors of Eucalyptus essential oils

Vapors of essential oils extracted from various species of Eucalyptus (E. gunnii, E. tereticornis, E. grandis, E. camaldulensis, E. dunnii, E. cinerea, E. saligna, E. sideroxylon, E. globulus ssp. globulus, E. globulus ssp. maidenii, E. viminalis and the hybrids E. grandis × E. tereticornis and E. grandis × E. camaldulensis) and their major components were found to be toxic to Aedes aegypti adults, the yellow fever mosquito.An aliquot of each oil was placed in a cylindrical test chamber and the number of knocked-down mosquitoes was recorded as function of time. Knockdown time 50% was then calculated. Results showed that E. viminalis had the fastest knockdown time at of 4.2 min, on the same order as dichlorvos, a standard knockdown agent. A correlation was observed between the content of 1,8-cineole in the Eucalyptus essential oils and the corresponding toxic effect.The correlation between KT₅₀ values and calculated vapor pressures of the essential oil components showed that the fumigant activity of simple organic compounds in insects is correlated with their volatility.     

Individual and combined effects of Bacillus thuringiensis var. israelensis, temephos and Leptolegniachapmanii on the larval mortality of Aedes aegypti

Larvicidal effects of interaction between Bacillus thuringiensis var. israelensis (Bti), temephos and Leptolegnia chapmanii zoospores on larvae of Aedes aegypti were determined under laboratory and seminatural conditions. In laboratory bioassays, two concentrations of Bti (0.012, 0.027 ppm), two of temephos (0.00035, 0.001 ppm), and a single concentration of L. chapmanii zoospores (6.1 × 10⁴ zoospores ml⁻¹) were evaluated. Trials under field-like conditions were performed in a single container and then placed either in the shade or in direct exposure to sunlight. We evaluated concentrations of Bti and temephos at 3-fold those normally used in laboratory tests: 0.09 and 0.003 ppm, respectively, plus 1.8 × 10⁵ zoospores ml⁻¹ of L. chapmanii. The combined effect of sublethal concentrations of Bti, temephos, and L. chapmanii zoospores thus indicated that this fungus is not inhibited by the larvicides and also demonstrated the synergistic effect of the action of L. chapmanii when used together with Bti and temephos.     

Mating systems in representatives of Parmeliaceae, Ramalinaceae and Physciaceae (Lecanoromycetes, lichen-forming ascomycetes)

The progeny of meiosis of eight Parmeliaceae, two Ramalinaceae and seven Physciaceae were subjected to fingerprint analysis using RAPD-PCR applied to single spore isolates. The sample set included common and widespread rarely fertile species (Parmelia sulcata, Pseudevernia furfuracea, Physcia tenella), local to common, infrequently fertile species (Melanelixia glabra, Parmelina tiliacea, Xanthoparmelia conspersa, X. stenophylla, Anaptychia runcinata, Diploicia canescen, Physconia distorta), local to rare, infrequently or regularly fertile species with declining distributions (Parmelina carporrhizans, P. quercina, Ramalina fastigiata, R. fraxinea, Anaptychia ciliaris), and local to common, regularly fertile species (Physcia aipolia, P. stellaris). All species turned out to be heterothallic, polymorphisms among RAPD markers ranging from 10–87 %. The significance of these findings for population genetics and conservation biology, and potential reasons for infrequent ascoma formation in some of the species are discussed.     

Suppression of RNA interference increases alphavirus replication and virus-associated mortality in Aedes aegypti mosquitoes

Background  

Arthropod-borne viruses (arboviruses) can persistently infect and cause limited damage to mosquito vectors. RNA interference (RNAi) is a mosquito antiviral response important in restricting RNA virus replication and has been shown to be active against some arboviruses. The goal of this study was to use a recombinant Sindbis virus (SINV; family Togaviridae; genus Alphavirus) that expresses B2 protein of Flock House virus (FHV; family Nodaviridae; genus Alphanodavirus), a protein that inhibits RNAi, to determine the effects of linking arbovirus infection with RNAi inhibition.