Mosquito traps designed to capture Aedes aegypti (Diptera: Culicidae) females: preliminary comparison of Adultrap, MosquiTRAP and backpack aspirator efficiency in a dengue-endemic area of Brazil

In this report, the efficiency of Adultrap under field conditions is compared to a CDC backpack aspirator and to MosquiTRAP. An urban dengue-endemic area of Rio de Janeiro was selected to evaluate the efficiency of mosquito traps in capturing Aedes aegypti females. Adultrap and aspirator captured similar numbers of Ae. aegypti females, with the former showing high specificity to gravid individuals (93.6%). A subsequent mark-release-recapture experiment was conducted to evaluate Adultrap and MosquiTRAP efficiency concomitantly. With a 6.34% recapture rate, MosquiTRAP captured a higher mean number of female Ae. aegypti per trap than Adultrap (chi2 = 14.26; df = 1; p < 0,05). However, some MosquiTRAPs (28.12%) contained immature Ae. aegypti after 18 days of exposure in the field and could be pointed as an oviposition site for female mosquitoes. Both trapping methods, designed to collect gravid Ae. aegypti females, seem to be efficient, reliable and may aid routine Ae. aegypti surveillance.     

Field optimisation of MosquiTRAP sampling for monitoring Aedes aegypti Linnaeus (Diptera: Culicidae)

A sticky trap designed to capture gravid Aedes (Stegomyia) aegypti mosquitoes, MosquiTRAP, has been evaluated for monitoring this species in Brazil. However, the effects of trap densities on the capture rate of Ae. aegypti females and the sensitivity of vector detection are still unknown. After a preliminary study has identified areas of high and low female mosquito abundance, a set of experiments was conducted in four neighbourhoods of Belo Horizonte (state of Minas Gerais, Brazil) using densities of 1, 2, 4, 8, 16, 32 and 64 traps per block. Trap sensitivity (positive MosquiTRAP index) increased significantly when 1-8 MosquiTRAPs were installed per block in both high and low abundance areas. A strong fit was obtained for the total number of mosquitoes captured with increasing trap densities through a non-linear function (Box-Lucas) (r2 = 0,994), which likely exhibits saturation towards an equilibrium level. The capacity of the Mean Female Aedes Index to distinguish between areas of high and low Ae. aegypti abundance was also investigated; the achieved differentiation was shown to be dependent on the MosquiTRAP density.     

Effects of variations in a formulated protein meal on the fecundity and fertility of female mosquitoes

Abstract. A formulated protein meal developed by Kogan (1990) for adult female Aedes aegypti mosquitoes was evaluated and modified to increase egg and pupal yield. A vigorous laboratory colony was maintained with the females fed exclusively on this dietary formula for about twenty-five generations over more than 2 years. Extra modifications were made to produce a diet suitable for Anopheles arabiensis and Anopheles stephensi females to produce eggs. Both formulations contain bovine albumin, haemoglobin and globulin in a ringer based solution, plus ATP as a phagostimulant for Ae.aegypti. Compared to Kogan's original, our Aedes formula doubled the production of pupae per female after a single meal, although the yield was still significantly lower than from mosquitoes fed on animal hosts or defibrinated pig blood. In varying the proportions of different constituents during attempts to optimize the formula, no relationship was found between total protein content (within the range 80–220 mg/ml) and fecundity, percentage hatch or pupal yield of Ae.aegypti. Equivocal results were found when an isoleucine supplement was added to the formula.     

Laboratory and field evaluation of the effects of the neonicotinoid imidacloprid on the oviposition response of Aedes (Stegomyia) aegypti Linnaeus (Diptera: Culicidae)

In this paper, we assessed the suitability of using the neonicotinoid imidacloprid with standard ovitraps by evaluating the ovicidal properties of imidacloprid and its influence on the oviposition response of gravid females of Aedes (Stegomyia) aegypti Linnaeus (Diptera: Culicidae). First, we calculated the imidacloprid lethal dose 99 (LD(99)) by exposing third instar larvae of the target species to different concentrations of the insecticide. Next, Ae. aegypti eggs were exposed to the imidacloprid LD(99) for 24 h and hatching inhibition was recorded. Finally, we investigated any potential repellent effect of the imidacloprid solution on the oviposition response of gravid Aedes females in field and laboratory conditions. The LD(99) obtained from larvae tests proved to be sufficient to keep any exposed eggs from hatching. No repellent effect was observed; females laid as many eggs in imidacloprid-treated ovitraps as in traps containing either clean water or temephos-treated water in both field and laboratory conditions. Our results indicate that imidacloprid is a suitable insecticide for treating ovitraps against Ae. aegypti.     

Horizontal transfer of the insect growth regulator pyriproxyfen to larval microcosms by gravid Aedes albopictus and Ochlerotatus triseriatus mosquitoes in the laboratory

The insect growth regulator (IGR) pyriproxyfen is highly active against mosquitoes (Diptera: Culicidae). Through continuous emersion of large larvae (instars 3-4) the concentration causing 50% inhibition of adult emergence (EI50) was determined as 0.200 p.p.b. for Aedes albopictus (Skuse) and 3.5 to 7 times less for Ochlerotatus triseriatus (Say): IE50 0.0288 p.p.b. As a possible method of application to larval microcosms of these species that oviposit in water containers and phytotelmata, the horizontal transfer of pyriproxyfen to larval microcosms by adult mosquitoes was evaluated under laboratory conditions. Gravid females were forced to walk on surfaces treated with pyriproxyfen (tarsal contact exposure) and then allowed to oviposit in larval microcosms. Using replicate bioassay cages, each with an oviposition container, and a factorial experimental design, we assessed Ae. albopictus for the effects of (i) pyriproxyfen concentration (0.2, 0.3 and 0.4 mg/cm2) contacted by gravid females, and (ii) the number of treated gravid females added to bioassay cages (one, three or five females/cage), on the mortality of larvae in oviposition containers. Only 0.2 mg/cm2 treatment rate was tested on Oc. triseriatus. A significant (P < 0.05) curvilinear response in inhibition of emergence (IE) was achieved on both species. Densities of one or three treated Oc. triseriatus females/cage yielded IE rates of only 21-27%, whereas five treated females/cage resulted in 70% inhibition. With Ae. albopictus, densities of three or five treated females/cage yielded 48-67% and 59-73% IE, respectively, whereas one treated female/cage gave only 4-30% inhibition. Use of IGR-treated oviposition containers to achieve horizontal transfer of pyriproxyfen to mosquito oviposition sites could be a field management technique based on mosquito biology and behaviour. In binary choice tests with Ae. albopictus, horizontal transfer of pyriproxyfen from a container with a treated ovistrip (0.3 or 0.4 mg/cm2) to an untreated microcosm resulted in 14-38% inhibition. In larval bioassays, pyriproxyfen activity declined markedly within 10 days. Forcibly exposing gravid female mosquitoes to pyriproxyfen-treated paper surface did not affect their fecundity. However, from the 1st to 2nd gonotrophic cycles the egg hatch rate declined by 30% (P < 0.05). Some variation of results could be due to interactions between females at the oviposition site, possibly causing disproportionate transfer of pyriproxyfen to larval microcosms. Comparative studies of the oviposition behaviour of each mosquito are warranted and would potentially provide information needed to improve the technique.     

Suboptimal Larval Habitats Modulate Oviposition of the Malaria Vector Mosquito Anopheles coluzzii

Selection of oviposition sites by gravid females is a critical behavioral step in the reproductive cycle of Anopheles coluzzii, which is one of the principal Afrotropical malaria vector mosquitoes. Several studies suggest this decision is mediated by semiochemicals associated with potential oviposition sites. To better understand the chemosensory basis of this behavior and identify compounds that can modulate oviposition, we examined the generally held hypothesis that suboptimal larval habitats give rise to semiochemicals that negatively influence the oviposition preference of gravid females. Dual-choice bioassays indicated that oviposition sites conditioned in this manner do indeed foster significant and concentration dependent aversive effects on the oviposition site selection of gravid females. Headspace analyses derived from aversive habitats consistently noted the presence of dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS) and 6-methyl-5-hepten-2-one (sulcatone) each of which unitarily affected An. coluzzii oviposition preference. Electrophysiological assays across the antennae, maxillary palp, and labellum of gravid An. coluzzii revealed differential responses to these semiochemicals. Taken together, these findings validate the hypothesis in question and suggest that suboptimal environments for An. coluzzii larval development results in the release of DMDS, DMTS and sulcatone that impact the response valence of gravid females.     

Influence of the spatial distribution of human hosts and large size containers on the dispersal of the mosquito Aedes aegypti within the first gonotrophic cycle

It is generally accepted that Aedes aegypti (L.) (Diptera: Culicidae) has a short dispersal capacity, and that displacement can be influenced by the availability of oviposition sites in the surroundings of emergence or release sites. In the present article, we observed the influence of spatial heterogeneity of large containers and human hosts on the cumulative flight direction of Ae. aegypti females during the first gonotrophic cycle, testing the hypothesis that they aggregate in resource-rich areas, i.e. where there are higher concentrations of large containers and/or humans per habitation. We analysed data from pupal surveys and mark-release-recapture experiments (non-blood-fed females were released) carried out in two dengue endemic neighbourhoods of Rio de Janeiro, Brazil: Tubiacanga (a suburb, with a human density of 337 inhabitants/ha) and Favela do Amorim (a slum, with a human density of 901 inhabitants/ha). In both areas, host-seeking females of three different release cohorts showed an overall non-uniform and extensive dispersal from their release point within 1–2 days post-release. At 4–5 days post-release, when many of the released females would be expected to be gravid, in Tubiacanga most mosquitoes were collected in areas with a relatively higher density of containers/premise, independently of the density of residents/house, whereas in Favela do Amorim, almost half of the captured mosquitoes were collected in relatively resource-poorer areas. Although Ae. aegypti dispersal patterns varied between sites, overall the distances travelled from the release point and the cumulative flight directions were correlated with the density of containers and hosts, more markedly in Tubiacanga than in Favela do Amorim.     

Experience‐ and age‐mediated oviposition behaviour in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti)

In repeated behaviours such as those of feeding and reproduction, past experiences can inform future behaviour. By altering their behaviour in response to environmental stimuli, insects in highly variable landscapes can tailor their behaviour to their particular environment. In particular, female mosquitoes may benefit from plasticity in their choice of egg‐laying site as these sites are often temporally variable and clustered. The opportunity to adapt egg‐laying behaviour to past experience also exists for mosquito populations as females typically lay eggs multiple times throughout their lives. Whether experience and age affect egg‐laying (or oviposition) behaviour in the mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) was assessed using a wind tunnel. Initially, gravid mosquitoes were provided with a cup containing either repellent or well water. After ovipositing in these cups, the mosquitoes were blood‐fed and introduced into a wind tunnel. In this wind tunnel, an oviposition cup containing repellent was placed in the immediate vicinity of the gravid mosquitoes. A cup containing well water was placed at the opposite end of the tunnel so that if the females flew across the chamber, they encountered the well water cup, in which they readily laid eggs. Mosquitoes previously exposed to repellent cups became significantly more likely to later lay eggs in repellent cups, suggesting that previous experience with suboptimal oviposition sites informs mosquitoes of the characteristics of nearby oviposition sites. These results provide further evidence that mosquitoes modify behaviour in response to environmental information and are demonstrated in a vector species in which behavioural plasticity may be ecologically and epidemiologically meaningful.     

The influence of larval density, food stress, and parasitism on the bionomics of the dengue vector Aedes aegypti (Diptera: Culicidae): implications for integrated vector management

New larval control strategies for integrated vector management of Aedes aegypti are in high demand, including the use of biological control agents. Exposure of Aedes aegypti to parasites, starvation, and overcrowded conditions during larval development reduces the probability of survival to eclosion, can directly affect fitness parameters such as adult size and fecundity, and can affect the size, provisioning, and viability of eggs produced by females. We compared these parameters after exposing larvae to 1) abundant food at low larval densities, 2) food deprivation and high larval density, and 2) infection with the endoparasite Plagiorchis elegans, an entomopathogenic digenean trematode. Female mosquitoes that eclosed from larval conditions of starvation and overcrowding were smaller and laid fewer and smaller eggs than controls. The proportion of females to complete an oviposition cycle was reduced in the P. elegans-infected treatment group. Parasite load was negatively correlated with wing length and egg size. Infection of Ae. aegypti with P. elegans has sublethal effects and may reduce population-level reproductive output, but one-time low-density P. elegans exposure does not have sufficient effect on Ae. aegypti fitness parameters to be considered a viable biocontrol option.     

Humoral control of pre-oviposition behaviour in the mosquito,Aedes aegypti

Pre-oviposition behaviour, the attraction of mosquitoes to oviposition-site stimuli, is induced by a haemolymph-borne substance. A large proportion of gravid mosquitoes was attracted to a solution of methyl propionate in a laboratory olfactometer, but non-gravid females were also attracted if they were first injected with haemolymph from gravid females. Ovariectomized blood-fed mosquitoes failed to respond, but the reimplantation of denervated ovaries or as few as six mature follicles still triggered pre-oviposition after a blood meal.     

Larvicidal and smoke repellency effect of Toddalia asiatica and Aegle marmelos against the dengue vector, Aedes aegypti (Insecta: Diptera: Culicidae)

Investigations were made to evaluate the larvicidal activity and smoke repellent potential of Toddalia asiatica (L.) and Aegle marmelos at different concentrations (20, 40, 60, 80 and 100 p.p.m.) against first to fourth instar larvae and pupae of Aedes aegypti (L.). The LC₅₀ of A. marmelos and T. asiatica against first instars was 50.960 and 47.893 p.p.m., respectively; against second instars, 52.979 and 50.922; third instars, 56.653 and 54.461; and fourth instars, 60.778 and 61.278. The LC₅₀ and LC₉₀ for A. marmelos and T. asiatica against pupae were 56.634 and 112.992, and 53.64 and 116.22 p.p.m., respectively. The smoke toxicity of T. asiatica against Ae. aegypti was greater than that of A. marmelos . Smoke-exposed gravid females oviposited fewer eggs when compared to those that were not exposed to smoke. Smoke-exposed females hatched a lower percentage of eggs compared to unexposed females.     

Physiological state of Aedes (Stegomyia) aegypti mosquitoes captured with MosquiTRAPs in Mirassol, S?o Paulo, Brazil.

We examined the best location in a house to install MosquiTRAPs, sticky ovitraps that attract and capture Aedes aegypti females and investigated the physiological state of captured female mosquitoes. The study was performed in a twenty-block area in Mirassol, S?o Paulo State, Brazil, in which five blocks were randomly chosen for MosquiTRAP installation. In each block, four houses were selected for the installation of eight traps: four indoors (bedroom, living room, bathroom, and kitchen) and four outdoors in the shade (two at the front of the house and two in the backyard). These houses were visited over an eight-week period. The outdoor MosquiTRAPs captured five times more females than indoor traps and appeared to be the best places to install MosquiTRAPs. There were no significant differences among indoor sites or among outdoor sites with respect to the number of females captured. The capacity of the MosquiTRAP to capture a large number of gravid Ae. aegypti females reinforces its potential as an entomological surveillance tool in dengue control programs.     

Methyl 3-chloro-4-methoxybenzoate, a new candidate semiochemical inhibiting copulation behaviour of Ixodes ricinus (L.) males

Abstract.  Females of Ixodes ricinus (L. 1758) emit several semiochemicals influencing copulatory behaviour in males. One of these candidate compounds, methyl 3-chloro-4-methoxybenzoate, is emitted exclusively by engorged females, and shows a copulation inhibiting effect in males. In Y-tube experiments, the synthetic analogue of this compound at a concentration of 100 µg/mL in ethanol, attracted males in only 25.6% of tests, whereas it repelled them in 74.4% of cases. However, unengorged females do not respond to this substance at all. The number of contacts between males and unengorged females treated with methyl 3-chloro-4-methoxybenzoate (250 µg/mL in ethanol, 1 µL per female) decreases to 28% compared to a control level of 80%, and the percentage of copulating pairs decreases to 12% compared to control (49%) over 2 h. Possibly, a fertilized engorged female with mature eggs uses this semiochemical to send a signal to males indicating that another copulation is no longer needed.     

Aedes aegypti oviposition in response to NPK fertilizers

Fertilizers are mineral associations intended to bring to the plants nutritive complements necessary to their growth. Modern fertilizers (NPK) combine the three basic elements which are the nitrogen (N), phosphorus (P) and potassium (K). In this study, we investigated in tunnel apparatus the influence of aqueous solutions containing low, moderate and high concentrations of NPK on the oviposition of Aedes aegypti. The results showed that the solutions containing moderate concentrations (NK = 17-33 mg/l and P = 23-47 mg/l) attracted significantly more gravid females than distilled water (P < 0.001). Conversely, the solutions containing either low or high concentrations of NPK (NK = 8 mg/l and P = 12 mg/l; NK = 50 mg/I and P = 70 mg/l) did not induce significant attraction (P > 0.05). These findings suggest that NPK fertilizers may influence the egg-laying behaviour of Ae. aegypti in field situations.     

Attraction of mosquitoes to volatiles associated with blood.

Responses of the mosquitoes Aedes aegypti, Culex quinquefasciatus, and Culex nigripalpus to volatiles and compounds associated with bovine and avian blood that were presented in collagen membranes were evaluated in olfactometer and landing assays. The presence of attractants produced by blood was supported by more attraction of all species to blood than water controls in the olfactometer. Females of Ae. aegypti and Cx. quinquefasciatus were more attracted to bovine blood than to avian blood, but there was no difference in Cx. nigripalpus responses. In landing assays, significantly more females of all species landed on casings with blood than on water controls. There was no difference in landing of Ae. aegypti on bovine or avian blood. However, significantly more females of Cx. quinquefasciatus and Cx. nigripalpus landed on avian blood compared to bovine blood. Blood presented in collagen casings was an effective method for evaluating in-flight attraction and landing in all three species. In the olfactometer, several individual compounds elicited attraction in all species, but none were as attractive as blood for all species. In landing assays, several organic acids and sulfides elicited landing, with Ae. aegypti responding to the greatest number of compounds. These assay methods are effective for evaluation of volatile compounds from blood, and although responses were obtained to several compounds, none were as effective as blood in the olfactometer and landing assays.     

Serratia odorifera a Midgut Inhabitant of Aedes aegypti Mosquito Enhances Its Susceptibility to Dengue-2 Virus

Mosquito midgut plays a crucial role in its vector susceptibility and pathogen interaction. Identification of the sustainable microflora of the midgut environment can therefore help in evaluating its contribution in mosquito-pathogen interaction and in turn vector competence. To understand the bacterial diversity in the midgut of Aedes aegypti mosquitoes, we conducted a screening study of the gut microbes of these mosquitoes which were either collected from fields or reared in the laboratory "culture-dependent" approach. This work demonstrated that the microbial flora of larvae and adult Ae. aegypti midgut is complex and is dominated by Gram negative proteobacteria. Serratia odorifera was found to be stably associated in the midguts of field collected and laboratory reared larvae and adult females. The potential influence of this sustainable gut microbe on DENV-2 susceptibility of this vector was evaluated by co-feeding S. odorifera with DENV-2 to adult Ae. aegypti females (free of gut flora). The observations revealed that the viral susceptibility of these Aedes females enhanced significantly as compared to solely dengue-2 fed and another gut inhabitant, Microbacterium oxydans co-fed females. Based on the results of this study we proposed that the enhancement in the DENV-2 susceptibility of Ae. aegypti females was due to blocking of prohibitin molecule present on the midgut surface of these females by the polypeptide of gut inhabitant S. odorifera.     

Genetic control of feeding preferences in the mosquitoes Aedes (Stegomyia) simpsoni and aegypti

ABSTRACT. The biting rate of a non-anthropophilic (Bwayise) population of Aedes simpsoni was found to be approximately 0.3 mosquitoes per catcher per hour, whereas that of an anthropophilic (Bwamba) population was approximately 101 per catcher per hour. Population density indices, as determined by the number of pupae per wet plant axil, were 0.70 in Bwayise and 1.00 in Bwamba. The big difference in anthropophilic behaviour between these populations was therefore unlikely to be derived from this small population difference. Larval density was higher at Bwamba than at Bwayise, but isolation or crowding of the larvae in the laboratory did not affect the biting behaviour of adult Ae. simpsoni. Laboratory studies also failed to confirm field observations that temperature might play a part in determining anthropophily and non-anthropophily in this species. In choice-chamber landing tests, using a rat and a human hand, Ae. simpsoni females derived from wild larvae and reared in the laboratory showed that 83% of the Bwamba strain landed on man, whereas only 38% of the Bwayise strain did so. In Aedes aegypti , 71% of a long-established laboratory strain (Ilobi) landed on man, whereas 47% of a relatively non-anthropophilic wild (Kampala) strain did so. These preferences persisted in culture. Selective breeding increased the preference for the rodent significantly in the Kampala strain of Ae. aegypti , but had no significant effect on the Ilobi strain. Crossbreeding showed that the F₁ and F₂ hybrids between the anthropophilic and non-anthropophilic strains were intermediate in their preference between the parental pure bred strains; the reciprocal crosses were not significantly different from each other. The behaviour of the backcross progenies, at least in Ae. aegypti , appeared to indicate that the genotype of the male parent might be the main determining factor.     

Identity and transfer of male reproductive gland proteins of the dengue vector mosquito, Aedes aegypti: potential tools for control of female feeding and reproduction

Male reproductive gland proteins (mRGPs) impact the physiology and/or behavior of mated females in a broad range of organisms. We sought to identify mRGPs of the yellow fever mosquito, Aedes aegypti, the primary vector of dengue and yellow fever viruses. Earlier studies with Ae. aegypti demonstrated that "matrone" (a partially purified male reproductive accessory gland substance) or male accessory gland fluid injected into virgin female Ae. aegypti affect female sexual refractoriness, blood feeding and digestion, flight, ovarian development, and oviposition. Using bioinformatic comparisons to Drosophila melanogaster accessory gland proteins and mass spectrometry of proteins from Ae. aegypti male accessory glands and ejaculatory ducts (AG/ED) and female reproductive tracts, we identified 63 new putative Ae. aegypti mRGPs. Twenty-one of these proteins were found in the reproductive tract of mated females but not of virgin females suggesting that they are transferred from males to females during mating. Most of the putative mRGPs fall into the same protein classes as mRGPs in other organisms, although some appear to be evolving rapidly and lack identifiable homologs in Culex pipiens, Anopheles gambiae, and D. melanogaster. Our results identify candidate male-derived molecules that may have an important influence on behavior, survival, and reproduction of female mosquitoes.