Ovitrap surveys of dengue vector mosquitoes in Chiang Mai, northern Thailand: seasonal shifts in relative abundance of Aedes albopictus and Ae. aegypti

Aedes aegypti (L.) and Aedes albopictus (Skuse) were surveyed using ovitraps in residential areas in Chiang Mai, northern Thailand. Egg populations (both species inclusive) remained low in the dry season, but increased/decreased exponentially during the first/latter half of the rainy season, respectively. This seasonal pattern was similar to the seasonal distribution of dengue haemorrhagic fever cases in the area. During the dry season (November-March) Ae.aegypti was dominant in urban and indoor ovitraps. With onset of the rainy season in April, relative abundance of Ae.albopictus increased in rural and outdoor ovitraps. Ae.albopictus displaced Ae.aegypti in the latter half of the rainy season in the rural area. Possible mechanisms to account for this seasonal decline of Ae.aegypti and reciprocal fluctuations in relative abundance of Ae.albopictus are discussed in relation to food availability for larvae in container habitats.     

Dissection of midgut and salivary glands from Ae. aegypti mosquitoes

The mosquito midgut and salivary glands are key entry and exit points for pathogens such as Plasmodium parasites and Dengue viruses. This video protocol demonstrates dissection techniques for removal of the midgut and salivary glands from Aedes aegypti mosquitoes.     

Duration and dose-dependency of female sexual receptivity responses to seminal fluid proteins in Aedes albopictus and Ae. aegypti mosquitoes

Male mosquitoes transfer seminal fluid proteins (hereafter ‘SFPs’) during mating. These proteins can have profound effects on female behavior in the yellow fever mosquito Aedes aegypti and the Asian tiger mosquito Aedes albopictus. SFPs are thought to be responsible for female refractoriness to mating in both species. However, only limited information is available about the duration of induced refractoriness or the quantity of SFPs required to be effective in Ae. albopictus. Here, we tested the duration of the effect of SFPs on female refractory behavior for both Aedes species. Additionally, we determined the lowest SFP dose required to induce female refractory behavior in Ae. aegypti. Virgin females were injected intra-thoracically with doses ranging from 0.25 to 0.008 equivalents of one male’s SFP amount. Our results demonstrate high sensitivity of female Ae. aegypti and Ae. albopictus to SFPs of their own species, with the majority of females becoming refractory at doses ? 0.031 male-equivalents after injection into the hemocoel. This effect was long-lasting in both species; none of the injected females were inseminated when presented with males of their own species 30 to 34 days post-injection, whereas most saline-injected control females mated at this time point. These results will aid future work to characterize individual SFPs involved in post-mating refractoriness in these two species. Moreover, they show that as is the situation in the mosquito Anopheles gambiae, and unlike Drosophila melanogaster, sperm are not required for the maintenance of a sexual refractoriness response in Ae. aegypti and Ae. albopictus.     

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.     

Egg hatching of mosquitoes Aedes caspius and Ae. vittatus stimulated by water vibrations

Simple laboratory experiments with eggs of wild-caught Aedes caspius (Pallas) and Aedes vittatus (Bigot) mosquitoes (Diptera: Culicidae) demonstrated that egg hatch rates (i.e. percentage hatching daily) increased significantly in response to water vibrations caused by finger-tip drumming on containers of submerged eggs. When disturbed by daily vibrations for only 30 s, eggs hatched sooner than when they remained flooded or were periodically drained and reflooded without agitation. For autogenous Ae. caspius from saltmarsh, egg hatch rates were three- to four-fold greater for groups of 50 eggs than for solitary eggs, possibly due to chemicals (functional kairomones) emanating from other eggs or larvae. For anautogenous Ae. vittatus from freshwater rock-pools, tapping the container daily (for 30 s each 2 h in 8 h) hatched 93% of eggs, compared with only 42% hatch of eggs agitated only when first flooded or 55% hatch of eggs dried and reflooded three times during 17 days without other disturbance. It is concluded that, apart from flooding and other factors, vibrations - simulating the patter of rainfall provide a very significant hatching stimulus for Aedes eggs.     

Consequences of the introduction of the planarian Girardia anceps (Tricladida: Dugesiidae) in artificial containers with larvae of the mosquitoes Aedes aegypti and Culex pipiens (Diptera: Culicidae) from Argentina

Aedes aegypti and Culex pipiens are container-dwelling mosquito species that are vectors of important diseases to man, such as dengue and lymphatic filariasis, respectively. Predators of these pests are an interesting alternative to be incorporated to biological control measures. We tested the consequences of introducing individuals of Girardia anceps, a native freshwater flatworm species, within artificial water containers where larvae of these mosquitoes thrive. Our goals were to ascertain if mosquito species, density of larvae (high or low), type of water container (tires or ovitraps), and presence or absence of planarians affected mosquito survivorship (measured as number of individuals reaching the pupa stage) in manipulated artificial containers. Furthermore, we monitored ovitraps in the field along several months in order to explore the long-term effect of the presence of planarian on the colonization of these containers by feral mosquitoes under natural conditions. We found that the presence of planarians reduced the number of mosquitoes reaching pupation and that such reduction depends on the initial density of larvae. Reduction of populations of A. aegypti was high along the breeding season of this mosquito, being the effect less evident in C. pipiens. G. anceps could be an agent of control against container-breeding mosquitoes if its release in small water containers is complemented with other suitable management strategies.     

The relative importance and distribution of Aedes polynesiensis and Ae.aegypti larval habitats in Samoa

Abstract. In preparation for a Filariasis Control programme in Samoa, during 1978 monthly larval surveys of the vector mosquito Aedes polynesiensis were carried out in four study villages in the main island of Upolu. A more extensive survey of larval habitat distribution was then made in twenty-two villages of Upolu and eighteen of Savai'i island, to determine the importance of habitat types according to their abundance, volume of water and whether their productivity was permanent or seasonal. Ae.aegypti larval densities and habitat distribution were also monitored and the occurrence of predatory Toxorhynchites amboinensis larvae in northern Upolu was recorded from forty-one collections. Aedes Breteau and container indices fluctuated with the pattern of rainfall in two coastal villages and an inland bush village, but not in a coconut plantation community. The five main Aedes larval habitat types encountered were: 200 litre water-storage drums, discarded tins and bottles, coconut shells, automobile tyres and treeholes. Aedes immatures occurred perennially in drums and tree holes, but breeding discontinued in tins, bottles and coconut shells during the driest month of July. For Ae.polynesiensis in Upolu the Breteau and container indices of 104.5 ± SD 80.9 and 35.3 ± 12.4 respectively were significantly higher than those in Savai'i: 33.1 ±25.0 and 24.3 ±20.0 respectively. Likewise for Ae.aegypti the Breteau and container indices of 50.8 ± 32.5 and 23.9 ± 15.6, respectively, were also significantly higher than those in Savai'i: 12.7 ± 17.1 and 9.4 ±13.2 respectively. Habitat types greater or lesser importance were determined by plotting the percentage of each type of container utilized for Aedes breeding against the percentage of each type amongst all larva-positive containers. Ae.polynesiensis preferred tree-holes but not water-storage drums. Ae.aegypti preferred drums and tyres; mixed populations of larvae of both species were commonest in these two types of habitat. Ae.polynesiensis occurred in every village. Ae.aegypti was encountered in all twenty-two villages surveyed in Upolu and nine of eighteen villages in Savai'i. Total larval surveys revealed that drums and tree-holes contained the highest numbers of Aedes larvae. The study provided criteria for planning a control programme.     

Characterization of hemocytes from the mosquitoes Anopheles gambiae and Aedes aegypti

Hemocytes are an essential component of the mosquito immune system but current knowledge of the types of hemocytes mosquitoes produce, their relative abundance, and their functions is limited. Addressing these issues requires improved methods for collecting and maintaining mosquito hemocytes in vitro, and comparative data that address whether important vector species produce similar or different hemocyte types. Toward this end, we conducted a comparative study with Anopheles gambiae and Aedes aegypti. Collection method greatly affected the number of hemocytes and contaminants obtained from adult females of each species. Using a collection method called high injection/recovery, we concluded that hemolymph from An. gambiae and Ae. aegypti adult females contains three hemocyte types (granulocytes, oenocytoids and prohemocytes) that were distinguished from one another by a combination of morphological and functional markers. Significantly more hemocytes were recovered from An. gambiae females than Ae. aegypti. However, granulocytes were the most abundant cell type in both species while oenocytoids and prohemocytes comprised less than 10% of the total hemocyte population. The same hemocyte types were collected from larvae, pupae and adult males albeit the absolute number and proportion of each hemocyte type differed from adult females. The number of hemocytes recovered from sugar fed females declined with age but blood feeding transiently increased hemocyte abundance. Two antibodies tested as potential hemocyte markers (anti-PP06 and anti-Dox-A2) also exhibited alterations in staining patterns following immune challenge with the bacterium Escherichia coli.     

Molecular analysis of mitochondrial cytochrome oxidase I gene of Aedes aegypti L. mosquitoes

Aedes aegypti is the most important arboviral vector worldwide. Recent studies reported that genetic variations and gene flow among same mosquito species is responsible for different disease transmission rate. Hence, to understand the relationship between genetic diversity and disease transmission potential, study on genetic variations among mosquito populations is essential. The aim of present study was to investigate the genetic variations of Ae. aegypti targeting COI gene from nine villages of Jalna District, Maharashtra and three laboratory strains originated from Aurangabad, Delhi and transgenic OX513A strain imported from OXITEC, UK. OX513A strain consists of a self-limiting dominant lethal gene construct intended for its use in suppression of Ae. aegypti population by sustained male adult releases in the environment. Mosquito eggs from field and laboratory strains were reared to adults and identified on the basis of morphological characteristics followed by COI gene sequence. Result of MSA and haplotype analysis revealed low genetic variations among field samples and Aurangabad strain, belonged to two haplotypes (H1 and H2) except Ramkheda village represented by separate haplotype H3. Other laboratory DEL strain and transgenic OX513A have great genetic variability to all isolates and have a separate haplotypes H4 and H5. Similar results were observed in phylogenetic analysis. Our observation of phylogenies revealed close relationship among the DEL and transgenic strain OX513A with few Indian and worldwide isolates. The information on genetic variability of mosquito population could help to understand and design the strategies for risk mitigation and effective implementation of new vector control tools like genetically modified mosquitoes.     

Container-breeding mosquitoes and predator community dynamics along an urban-forest gradient: The effects of habitat type and isolation

Environmental disturbances such as deforestation, urbanization or pollution have been widely acknowledged to play a key role in the emergence of many infectious diseases, including mosquito-borne viruses. However, we have little understanding of how habitat isolation affects the communities containing disease vectors. Here, we test the effects of habitat type and isolation on the colonization rates, species richness and abundances of mosquitoes and their aquatic predators in water-filled containers in northwestern Thailand. For eight weeks water-filled containers were monitored in areas containing forest, urban and agricultural habitats and mixtures of these three. Mosquito larvae of the genera Aedes and Culex appeared to be differentially affected by the presence of the dominant predator; Toxorhynchites splendens (Culicidae). Therefore, a predation experiment was conducted to determine predator response to prey density and its relative effects on different mosquito prey populations. Colonization rates, species richness and abundances of mosquito predators were strongly related to forest habitat and to the distance from other aquatic habitats. Areas with more tree cover had higher predator species richness and abundance in containers. Containers that were close to surface water were more rapidly colonized than those further away. In all habitat types, including urban areas, when predators were present, the number of mosquito larvae was much lower. Containers in urban areas closer to water-bodies, or with more canopy cover, had higher predator colonization rates and species richness. T. splendens (Culicidae) preyed on the larvae of two mosquito genera at different rates, which appeared to be related to prey behaviour. This study shows that anthropogenic landscape modification has an important effect on the natural biological control of mosquitoes. Vector control programmes and urban planning should attempt to integrate ecological theory when developing strategies to reduce mosquito populations. This would result in management strategies that are beneficial for both public health and biodiversity.     

The role of olfaction in host preference by Aedes (Stegomyia) simpsoni and Ae. aegypti

Host preference in females of the mosquitoes Aedes simpsoni (Theo.) and Aedes aegypti (L) appears to be based largely on responses to specific odours at short range, with the site of olfaction being the antennae. In blank Y-olfactometer tests, 48 % of an anthropophilic (llobi) strain of Ae. aegypti responded to one arm and 52% to the other. However, presented with a man's hand in one arm of the olfacto-meter and a whole rat, Arvicanthis niloticus , in the other, 65% responded to man and 35% to the rat. With the same strain in a blank Gouck's type of olfactometer, the response was very poor, only 1 % of the mosquitoes in the olfactometer responded at all, again with no preference for either side. On the introduction of the same hosts into the Gouck's olfactometer, the total response was c. 31 %, of which 72 % responded to man and 28 % to the rat. Elimination of some visual cues affected neither the total response nor the preference. Removing one antenna, reduced the preference for man from about 70 % in the normal mosquitoes to about 58%. The total response remained approximately the same as in the normal mosquitoes. Removing both antennae, in both Ae. aegypti and Ae. simpsoni , caused a sharp depression in the total response and in host discrimination.     

Effects of intraspecific larval competition on adult longevity in the mosquitoes Aedes aegypti and Aedes albopictus

Abstract Larval competition is common in container‐breeding mosquitoes. The impact of competition on larval growth has been thoroughly examined and findings that larval competition can lead to density‐dependent effects on adult body size have been documented. The effects of larval competition on adult longevity have been less well explored. The effects of intraspecific larval densities on the longevity of adults maintained under relatively harsh environmental conditions were tested in the laboratory by measuring the longevity of adult Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) that had been reared under a range of larval densities and subsequently maintained in high‐ or low‐humidity regimes (85% or 35% relative humidity [RH], respectively) as adults. We found significant negative effects of competition on adult longevity in Ae. aegypti, but not in Ae. albopictus. Multivariate analysis of variance suggested that the negative effect of the larval environment on the longevity of Ae. aegypti adults was most strongly associated with increased development time and decreased wing length as adults. Understanding how larval competition affects adult longevity under a range of environmental conditions is important in establishing the relationship between models of mosquito population regulation and epidemiological models of vector‐borne disease transmission.     

Horizontal and vertical dispersal of dengue vector mosquitoes, Aedes aegypti and Aedes albopictus, in Singapore

To study the dispersal of dengue vector mosquitoes in Singapore, females of Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) were fed blood containing rubidium (Rb), which was detectable in their eggs by means of Graphite Furnace Atomic Absorption Spectrophotometry (GFAAS). Laboratory calibration of the Rb reading, for a range of egg numbers from Rb-fed females, indicated a reasonably linear relationship and an unequivocal distinction between results with zero and one marked egg. Rb-marked female Aedes mosquitoes aged 3-5 days were released in semi-rural and urbanized parts of Singapore, with an array of ovitraps extending to a radius of 320 m from the release point. Subsequently, Rb-marked Aedes eggs were detected throughout the array, with similar distributions on each of the 4 days after release. More Rb was detected nearer the release point. However, when correction was made for the greater areas of zones further from the release point (and therefore presumably existence of more alternative oviposition sites), there were no significant differences in the numbers of marked eggs per ovitrap in the zones nearer or further from the release points. It is concluded that females of both these Aedes (Stegomyia) species could disperse easily and quickly throughout areas of radius 320 m in search of oviposition sites. This contrasts with the general belief that Ae. aegypti seldom flies more than 50 m and that control operations can safely be based on such an assumption. Releases on level 12 of a 21-storey apartment block, with ovitraps on each storey, showed similar easy and rapid dispersal to the top and bottom of the block.     

Field efficacy of triflumuron against Aedes and Culex mosquitoes in temperate Argentina

Aedes aegypti and Culex pipiens s.l. (Linnaeus, 1762 and 1758, respectively) (Diptera: Culicidae) are important vectors of diseases to humans and a growing public health concern. In order to contribute to the control of mosquito vectors by low environmental impact approaches we assessed the susceptibility of natural populations of container-breeding mosquitoes to triflumuron, an insect growth regulator, in temperate Argentina. A field trial was conducted to evaluate the efficacy of two doses (0.5 ppm and 1 ppm) of triflumuron (SC 48%) against natural populations of Ae. aegypti and Culex spp. immatures in flower vases of four cemeteries. The results demonstrated the susceptibility of both target mosquitoes to triflumuron in field conditions. For Ae. aegypti, dose-dependent reductions were achieved in the presence of pupae and the percentage of water-holding containers harbouring L3–4 and/or pupae, whereas the larvae abundance was equally reduced for both doses. For Culex spp., similar levels of reduction of larvae abundance and pupae presence were achieved with both doses. Significant effects on the response variables measured were recorded up to six to eight weeks post-intervention. Bimonthly applying 1 ppm triflumuron in the context of an integrated mosquito management should achieve a lasting control of Ae. aegypti and Culex spp. in small artificial containers with minimal environmental impacts.     

Electrophysiological and behavioural response of Aedes albopictus to n‐heinecosane,an ovipositional pheromone of Aedes aegypti

Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) are highly anthropophilic mosquito species and potential vectors of dengue and yellow fever. The location of suitable sites for oviposition requires a set of visual, tactile, and olfactory cues that influence females before they lay their eggs. In this study, the effect of n‐heneicosane, a recognized oviposition pheromone of Ae. aegypti, on the olfactory receptors of the antennae of Ae. aegypti and Ae. albopictus was studied using electroantennographic detection coupled to gas chromatography (GC‐EAD). A significant electroantennographic response to n‐heneicosane in adult females of both mosquito species was observed. In addition, gravid Ae. albopictus females laid more eggs in substrate treated with n‐heneicosane at 0.1, 1, or 10 p.p.m. than in the control, denoting oviposition attractancy. Conversely, at 30, 50, 100, and 200 p.p.m., more eggs were laid in the control substrate, indicating oviposition repellency. Analysis of the larval cuticle by GC and mass spectrometry confirmed the presence of n‐heneicosane in the cuticles of Ae. albopictus larvae. The species‐specific role of n‐heneicosane as an oviposition pheromone in Ae. aegypti and its significance as a behaviour modifier of Ae. albopictus in breeding sites is discussed.     

Ultrastructure and morphology of midgut visceral muscle in early pupal Aedes aegypti mosquitoes

These studies focus on the pupal Aedes aegypti midgut muscularis for the first 26 h following larval-pupal transition. The midgut muscularis of Ae. aegypti pupae during this first half of the pupal stadium is a grid of both circularly and longitudinally oriented muscle bands, arranged in a manner resembling that of the larvae. While many muscle bands exhibit signs of degeneration during the time period studied, not all bands degrade, nor is this degradation simultaneous. Band deterioration involves destruction of internal elements while the muscle fiber plasma membrane remains intact. Deterioration of contractile elements may involve proteosome-like structures and associated enzymes. Many features of the larval muscularis including cruciform cells, bifurcating circular bands, and bifurcating longitudinal bands of muscle are retained during the time period investigated. Neuromuscular junctions along some muscle bands are retained through at least 16 h into the pupal stadium. The selective nature of muscle fiber degradation, coupled with the retention of larval features and neural input, may allow for limited functionality of the muscularis during metamorphosis. Evidence of sexual dimorphism in the midgut muscularis of male and female Ae. aegypti pupae was not observed during the time period studied.     

Prevalence and seasonality of Ascogregarina culicis (Apicomplexa: Lecudinidae) in natural populations of Aedes aegypti (Diptera: Culicidae) from temperate Argentina

Ascogregarina infections from South America were recently documented in Brazil and Argentina. The aim of this study was to report our recent findings on the prevalence and seasonality of Ascogregarina culicis in Aedes aegypti adults from temperate Argentina. Between December 2003 and May 2005, 391 females of Ae. aegypti were captured in two areas of Greater Buenos Aires. Overall prevalence of A. culicis was 21.2% (83/391), and a significant difference was observed between both areas (28.4% vs. 8%). Infected Ae. aegypti were found from November to May, with highest values in March (24-37%). Parasite prevalence and host abundance showed similar seasonal patterns. Our observations suggest a widespread infestation of A. culicis among Ae. aegypti populations from temperate Argentina.     

Structure of host-odour plumes influences catch of Anopheles gambiae s.s. and Aedes aegypti in a dual-choice olfactometer

The effects of structure, concentration and composition of host‐odour plumes on catch of female Anopheles gambiae Giles sensu stricto and Aedes aegypti (L.) (Diptera: Culicidae) were investigated in a dual‐choice olfactometer. We demonstrate that the fine‐scale structure of host‐odour plumes modulates capture of An. gambiae and Ae. aegypti. In both species homogeneous skin‐odour plumes result in trap entry, whereas homogeneous CO₂ plumes reduce trap catch. Reduced trap catch also result from combining skin odour with a homogeneous CO₂ plume. Trap capture rates in homogeneous CO₂ plumes were concentration‐dependent and differed between the two species. Electric nets placed in front of the trap entrances intercepted mosquitoes before they could enter the traps. This showed that An. gambiae flew along CO₂ plumes, but did not enter the traps. Survivorship analysis of the trap‐entry times of Ae. aegypti indicated interactions between the time until capture and treatment. The assay's duration therefore can alter the distribution in a dual‐choice olfactometer.     

Adverse ffects of covert iridovirus infection on life history and demographic parameters of Aedes aegypti

Abstract Sublethal viral infections can cause changes in the body size and demography of insect vectors, with important consequences for population dynamics and the probability that individual mosquitoes will transmit disease. This study examined the effects of covert (sublethal) infection by Invertebrate iridescent virus 6 (IIV‐6) on the demography of female Aedes aegypti and the relationship between key life history parameters in covertly infected female insects compared with healthy (control) insects or non‐infected mosquitoes that had survived exposure to virus inoculum without becoming infected. Of the female mosquitoes that emerged following exposure to virus inoculum and were offered blood meals, 29% (43/150) proved positive for covert IIV‐6 infection. The net reproductive rate (R₀) of covertly infected females was 50% lower for infected females compared to control mosquitoes, whereas non‐infected exposed females had an R₀ approximately 15% lower than that of controls. Reproduction caused a significant decrease of about 13 days in mosquito longevity compared to females that did not reproduce (P < 0.001). Infected females lived 5–8 days less than non‐infected exposed females or controls, respectively (P = 0.028). Infected females and non‐infected exposed females both had significantly shorter wings than control insects (P < 0.001). There was a significant positive correlation between wing length and longevity in covertly infected female mosquitoes but not in control or non‐infected exposed mosquitoes. Longer lived females produced more eggs in all treatments. There were no significant correlations between body size and fecundity or the production of offspring. There was also no correlation between fecundity and fertility, suggesting that sperm inactivation was a more likely cause of decreased fertility in older mosquitoes than sperm depletion. We conclude that covert infection by iridescent virus is likely to reduce the vectorial capacity of this mosquito.