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.     

Risk of dengue occurrence based on the capture of gravid Aedes aegypti females using MosquiTRAP

We assessed the risk classification of dengue fever based on the capture of Aedes aegypti adults using MosquiTRAP, a type of sticky trap, in comparison with traditional larval infestation indices. A total of 27 MosquiTRAPs were installed, with one trap per block, and were inspected weekly between November 2008-February 2009. Infestation baseline data were obtained from a survey conducted prior to trap installation. The index generated by MosquiTRAP and house index (HI) classified the area "in alert situation". The set for risk of dengue occurrence proposed by the use of MosquiTRAP classify areas in the same way of the traditional HI.     

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.     

Evaluation of the sticky MosquiTRAP for detecting Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae) during the dry season in Belo Horizonte, Minas Gerais, Brazil

MosquiTRAP is a sticky trap specifically designed to capture gravid females of Aedes aegypti (L.) and allows the identification of the mosquito in the field during the inspection of the trap. This study aims to compare this sticky trap to larval and ovitrap surveys for field monitoring of A. aegypti during the dry season. The study was conducted from March to June of 2003 in 20 blocks of the district of Itapo?, Belo Horizonte, MG. The traps were monitored every week while the larval survey was conducted on a monthly basis. The larval index: Premise Index (PI) and Breteau Index (BI) had equal values throughout the experiment (1.72 in the first two months and zero in the last two). The container index (CI) during the first two months was 0.09 and 0.1%, respectively and zero in the last two. The Ovitrap Positive Index (OPI) ranged from 16.7 to 76.9%, and the MosquiTRAP Positive Index (MPI) ranged from 0 to 31.5%. The Egg Density Index (EDI) ranged from 26.6 to 82.8, while the Adult Density Index ranged from 0 to 1.6 throughout the experiment. Temperature and rainfall did not affect the Positive and Density Indices, although these environmental variables seemed to have affected the larvae indices. Although the MosquiTRAP caught a low number of Aedes mosquitoes during the study, it was more sensitive than the larval survey to detect the presence of Aedes mosquitoes.     

Development of a novel sticky trap for container-breeding mosquitoes and evaluation of its sampling properties to monitor urban populations of Aedes albopictus

Collection methods currently used for large-scale sampling of adult Stegomyia mosquitoes (Diptera: Culicidae) present several operational limitations, which constitute major drawbacks to the epidemiological surveillance of arboviruses, the evaluation of the impact of control strategies, and the surveillance of the spreading of allochthonous species into non-endemic regions. Here, we describe a new sticky trap designed to capture adult container-breeding mosquitoes and to monitor their population dynamics. We tested the sampling properties of the sticky trap in Rome, Italy, where Aedes (Stegomyia) albopictus is common. The results of our observations, and the comparison between sticky trap catches and catches made with the standard oviposition trap, are presented. The sticky trap collected significantly larger numbers of Ae. albopictus females than any other Culicidae species representing >90% of the total catches. A maximum of 83 An. albopictus females was collected in a single week. A high correlation (Pearson correlation coefficient r= 0.96) was found between the number of females and the number of eggs collected by the traps. The functional relationship between the number of eggs and the number of adult females was assessed by major axis regression fitted to log(1 +x)-transformed trap counts as y= 0.065 + 1.695x. Trap samples significantly departed from a random distribution; Taylor's power law was fitted to the trap samples to quantify the degree of aggregation in the catches, returning the equations s(2)= 2.401 m(1.325) for the sticky trap and s(2)= 13.068 m(1.441) for the ovitrap, with s(2) and m denoting the weekly catch variance and mean, respectively, indicating that eggs were significantly more aggregated than mosquitoes (P < 0.0001). Taylor's power law parameters were used to estimate the minimum number of sample units necessary to obtain sample estimates with a fixed degree of precision and sensitivity. For the range of densities encountered in our study area during the Ae. albopictus breeding season, the sticky trap was more precise and sensitive than the ovitrap. At low population densities (c. < 0.1 mosquito/trap), however, the ovitrap was more sensitive at detecting the presence of this species. Overall, our results indicate that our new model of sticky trap can be used to sample Ae. albopictus females in urban environments, and, possibly, other container-breeding Stegomyia mosquitoes (e.g. Aedes aegypti). The technical properties of the new trap are discussed with respect to its possible application in monitoring the population dynamics of container-breeding mosquitoes, in studying their bionomics, and in vector surveillance and, possibly, control.     

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.     

Wolbachia induces density-dependent inhibition to dengue virus in mosquito cells

Wolbachia is a maternal transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce viral interference and spread into mosquito vector population makes it possible to develop Wolbachia as a biological control agent for dengue control. While Wolbachia induces resistance to dengue virus in the transinfected Aedes aegypti mosquitoes, a similar effect was not observed in Aedes albopictus, which naturally carries Wolbachia infection but still serves as a dengue vector. In order to understand the mechanism of this lack of Wolbachia-mediated viral interference, we used both Ae. albopictus cell line (Aa23) and mosquitoes to characterize the impact of Wolbachia on dengue infection. A serial of sub-lethal doses of antibiotic treatment was used to partially remove Wolbachia in Aa23 cells and generate cell cultures with Wolbachia at different densities. We show that there is a strong negative linear correlation between the genome copy of Wolbachia and dengue virus with a dengue infection completely removed when Wolbacha density reaches a certain level. We then compared Wolbachia density between transinfected Ae. aegypti and naturally infected Ae. albopictus. The results show that Wolbachia density in midgut, fatbody and salivary gland of Ae. albopictus is 80-, 18-, and 24-fold less than that of Ae. aegypti, respectively. We provide evidence that Wolbachia density in somatic tissues of Ae. albopictus is too low to induce resistance to dengue virus. Our results will aid in understanding the mechanism of Wolbachia-mediated pathogen interference and developing novel methods to block disease transmission by mosquitoes carrying native Wolbachia infections.     

A comparison of larval,ovitrap and MosquiTRAP?surveillance for Aedes (Stegomyia) aegypti

In Brazil, the entomological surveillance of Aedes (Stegomyia) aegypti is performed by government-mandated larval surveys. In this study, the sensitivities of an adult sticky trap and traditional surveillance methodologies were compared. The study was performed over a 12-week period in a residential neighbourhood of the municipality of Pedro Leopoldo, state of Minas Gerais, Brazil. An ovitrap and a MosquiTRAP were placed at opposite ends of each neighbourhood block (60 traps in total) and inspections were performed weekly. The study revealed significant correlations of moderate strength between the larval survey, ovitrap and MosquiTRAP measurements. A positive relationship was observed between temperature, adult capture measurements and egg collections, whereas precipitation and frequency of rainy days exhibited a negative relationship.     

Preliminary data on the performance of Aedes aegypti and Aedes albopictus immatures developing in water-filled tires in Rio de Janeiro

A monthly survey of Aedes aegypti and Aedes albopictus immatures in discarded tires at a site in metropolitan Rio de Janeiro showed that Ae. albopictus was much more abundant in the rainy season, but Ae. aegypti abundance showed a less clear seasonal pattern. Pupal masses for Ae. albopictus showed a seasonal trend. In contrast, Ae. aegypti pupae did not show any clear trend in weight. Large Ae. albopictus pupae were found in the warmer months, when water volume was higher, pH lower and larval abundance lower. Further studies should be carried out to assess how seasonal variations in body size may impact vector competence of these species in Brazil.     

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.     

Parasitism of Ascogregarina taiwanensis and Ascogregarina culicis (Apicomplexa: Lecudinidae) in larvae of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) from Manaus, Amazon region, Brazil

The aim of the study was to determine the existence of Ascogregarina spp. in larvae of Aedes albopictus and Aedes aegypti collected in urban and suburban areas of Manaus, Amazon region, Brazil. Between May 2004 and July 2005, the mid-gut of 3rd and 4th instar larvae, collected in tire traps in six neighborhoods of Manaus, was examined for the presence of trophozoites of Ascogregarina. Coexistence of Ae. albopictus larvae infected by A. taiwanensis, and Ae. aegypti larvae by A. culicis, was detected in traps in the field. The percentage of Ae. albopictus larvae infected by A. taiwanensis ranged from 21% to 93.5% and of Ae. aegypti larvae infected by A. culicis from 22% to 95%. The mean infection intensity was similar in both species of Aedes. In traps located in Mauazinho, the replacement of Ae. aegypti by Ae. albopictus larvae was observed. In Manaus, Ae. albopictus larvae were parasitized by A. taiwanensis, and Ae. aegypti larvae by A. culicis. Infection rates were high when the species of Aedes were found separately.     

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.     

Mass trapping with MosquiTRAPs does not reduce Aedes aegypti abundance

The objective of this study was to evaluate the effectiveness of Aedes aegypti mass trapping using the sticky trap MosquiTRAP (MQT) by performing a cluster randomised controlled trial in Manaus, state of Amazonas, Brazil. After an initial questionnaire and baseline monitoring of adult Ae. aegypti abundance with BG-Sentinel (BGS) traps in six clusters, three clusters were randomly assigned to the intervention arm where each participating household received three MQTs for mass trapping during 17 months. The remaining three clusters (control arm) did not receive traps. The effect of mass trapping on adult Ae. aegypti abundance was monitored fortnightly with BGS traps. During the last two months of the study, a serological survey was conducted. After the study, a second questionnaire was applied in the intervention arm. Entomological monitoring indicated that MQT mass trapping did not reduce adult Ae. aegypti abundance. The serological survey indicated that recent dengue infections were equally frequent in the intervention and the control arm. Most participants responded positively to questions concerning user satisfaction. According to the results, there is no evidence that mass trapping with MQTs can be used as a part of dengue control programs. The use of this sticky trap is only recommendable for dengue vector monitoring.     

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 evaluation of a lethal ovitrap against dengue vectors in Brazil

Field evaluation of a "lethal ovitrap" (LO) to control dengue vector Aedes mosquitoes (Diptera: Culicidae), was undertaken in two Brazilian municipalities, Areia Branca and Nilopolis, in the State of Rio de Janeiro. The LO is designed to kill Aedes via an insecticide-treated ovistrip (impregnated with deltamethrin). In each municipality, the intervention was applied to a group of 30 houses (10 LOs/house) and compared to 30 houses without LOs in the same neighbourhood. Five LOs were put outside and five LOs inside each treated house. Three methods of monitoring Aedes density were employed: (i) percentage of containers positive for larvae and/or pupae; (ii) total pupae/house; (iii) total adult females/house collected by aspirator indoors. Weekly mosquito surveys began during the month before LO placement, by sampling from different groups of 10 houses/week for 3 weeks pre-intervention (i.e. 30 houses/month) and for 3 months post-intervention in both treated and untreated areas. Prior to LO placement at the end of February 2001, Aedes aegypti (L) densities were similar among houses scheduled for LO treatment and comparison (untreated control) at each municipality. Very few Ae. albopictus (Skuse) were found and this species was excluded from the assessment. Post-intervention densities of Ae. aegypti were significantly reduced for most comparators (P < 0.01), as shown by fewer positive containers (4-5 vs. 10-18) and pupae/house (0.3-0.7 vs. 8-10) at LO-treated vs. untreated houses, 3 months post-treatment at both municipalities. Numbers of adult Ae. aegypti females indoors were consistently reduced in LO-treated houses at Areia Branca (3.6 vs. 6.8/house 3 months post-intervention) but not at Niloplis (approximately 3/house, attributed to immigration). These results demonstrate sustained impact of LOs on dengue vector population densities in housing conditions of Brazilian municipalities.     

Productivity of natural and artificial containers for Aedes polynesiensis and Aedes aegypti in four American Samoan villages

Six mosquito species were identified in a survey of containers associated with 347 households in four villages in American Samoa. Aedes polynesiensis Marks (Diptera: Culicidae) and Aedes aegypti (L) were the most abundant species, representing 57% and 29% of the mosquitoes identified. Culex quinquefasciatus (Say), Culex annulirostris (Skuse), Aedes oceanicus (Belkin) and Toxorhynchites amboinensis (Doleschall) were also found. Aedes aegypti and Ae. polynesiensis showed distinct differences in their use of containers, preferring large and small containers, respectively. By contrast with previous studies, Ae. polynesiensis utilized domestic and natural containers with equal frequency, whereas Ae. aegypti continued to be found predominantly in domestic containers. Only 15% of containers holding immature mosquitoes included pupae and fewer than 10 Aedes spp. pupae were found in most containers with pupae. An estimated 2289 Ae. polynesiensis and 1640 Ae. aegypti pupae were found in 2258 containers. The presence of both species in the same container did not affect the mean density of either species for larvae or pupae. Glass jars, leaf axils, tree holes and seashells produced few Aedes spp. pupae in any of the study villages. Overall, 75% of Ae. polynesiensis pupae were found in buckets, ice-cream containers and tyres, with <7% being produced in natural containers, whereas 82% of Ae. aegypti pupae were found in 44-gallon (US) drums ( approximately 166L), buckets and tyres. Source reduction efforts targeting these container types may yield significant reductions in both Ae. polynesiensis and Ae. aegypti populations in American Samoa.     

Insecticide susceptible/resistance status in Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Thailand during 2003-2005

Susceptibility baselines and diagnostic doses of the technical grade insecticides deltamethrin, permethrin, fenitrothion, and propoxur were established based on Aedes aegypti (L.), Bora (French Polynesia), a reference susceptible strain. Field-collected Aedes mosquitoes from each part of Thailand were subjected to bioassay for their susceptibility to the diagnostic doses of each insecticide. Almost all Ae. aegypti collected were incipient resistant or resistant to deltamethrin and permethrin, except those from some areas of Songkhla (southern) and Phan district of Chiang Rai (northern) province. Susceptibility to fenitrothion was found in mosquitoes from Bangkok (central), Chonburi (eastern), Chiang Rai, Kanchanaburi (western), and Songkhla, whereas they were resistant in almost all areas of Nakhon Sawan (north central) and Nakhon Ratchasima (northeastern) provinces. Most of Ae. aegypti were susceptible to propoxur except those from Mae Wong, Nakhon Sawan province. Various levels of insecticide resistance and susceptibility in adjacent areas revealed a focal susceptible/resistance profile in the country. It could be noted that almost all of Ae. albopictus were susceptible to the insecticides tested at the same diagnostic doses. In conclusion, resistance to pyrethroids (permethrin and deltamethrin) has developed in Ae. aegypti in most of the collected areas, suggesting that an alternative choice of insecticide or other control measures should be applied.     

Discrepancies between Aedes aegypti identification in the field and in the laboratory after collection with a sticky trap

Currently, sticky traps are regularly employed to assist in the surveillance of Aedes aegypti infestation. We tested two alternative procedures for specimen identification performed by local health agents: directly in the field, as recommended by certain manufacturers, or after transportation to the laboratory. A total of 384 sticky traps (MosquiTRAP) were monitored monthly during one year in four geographically representative Brazilian municipalities. When the same samples were inspected in the field and in the laboratory, large differences were noted in the total number of mosquitoes recorded and in the number of specimens identified as Ae. aegypti by both procedures. Although field identification has the potential to speed vector surveillance, these results point to uncertainties in the evaluated protocol.     

Oral susceptibility to yellow fever virus of Aedes aegypti from Brazil

The oral susceptibility to yellow fever virus was evaluated in 23 Aedes aegypti samples from Brazil. Six Ae. aegypti samples from Africa, America and Asia were also tested for comparison. Mosquito samples from Asia showed the highest infection rates. Infection rates for the Brazilian Ae. aegypti reached 48.6%, but were under 13% in 60% of sample tested. We concluded that although the low infection rates estimated for some Brazilian mosquito samples may not favor the establishment of urban cycle of yellow fever in some parts of the country, the founding of Ae. aegypti of noteworthy susceptibility to the virus in cities located in endemic and transition areas of sylvatic yellow fever, do pose a threat of the re-emergence of the urban transmission of the disease in Brazil.     

Calibration and evaluation of field cage for oviposition study with Aedes (Stegomyia) aegypti female (L.) (Diptera: Culicidae)

Differences among results gathered from insect behavior studies conducted in laboratory and field situations are due to ambient variables that differ greatly between both environments. In laboratory studies the environmental conditions can be controlled whereas in field temperature, humidity and air velocity vary uncontrollably. The objective of this study was to calibrate and evaluate an experimental area (field cage) (14 x 7 x 3.5 m) subdivided into eight test cages (2.5 x 2.5 x 2 m) for use in behavioral oviposition tests of Aedes aegypti (L.) mosquitoes for developing a new methodology to assess attractants and oviposition traps. Test cage calibration involved: (1) minimal experiment duration tests; (2) optimal female release number per traps test and (3) trap placement tests. All tests used gravid A. aegypti females; 3-4 days post blood meal and the sticky trap MosquiTRAP to catch adults. Ninety percent of the females released were recaptured 2h after the beginning of the experiment, and this allowed up to 32 test repetitions/day to be conducted in the field cage. The minimum number of females necessary to conduct statistical analyses was 20 females/trap/test per cage. No significant difference was found in the behavioral response of gravid females to four different trap positions within test cages. Field trapping results with attractant were similar to those in the field cage. Therefore, the field cage could replace field trapping for evaluating at least mosquito traps and oviposition attractants for A. aegypti.