Luke P. Rapley

A lethal ovitrap-based mass trapping scheme for dengue control in Australia: II. Impact on populations of the mosquito Aedes aegypti

In Cairns, Australia, the impacts on Aedes aegypti L. (Diptera: Culicidae) populations of two types of ‘lure & kill’ (L&K) lethal ovitraps (LOs), the standard lethal ovitrap (SLO) and the biodegradable lethal ovitrap (BLO) were measured during three mass‐trapping interventions. To assess the efficacy of the SLO, two interventions (one dry season and one wet season) were conducted in three discrete areas, each lasting 4 weeks, with the following treatments: (i) SLOs (>200 traps, ∼4/premise), BG‐sentinel traps (BGSs; ∼15, 1/premise) and larval control (container reduction and methoprene treatment) and (ii) larval control alone, and (iii) untreated control. Female Ae. aegypti populations were monitored for 4 weeks pre‐ and post‐treatment in all three areas using BGSs and sticky ovitraps (SOs) or non‐lethal regular ovitraps (ROs). In the dry season, 206 SLOs and 15 BGSs set at 54 and 15 houses, respectively, caught and killed an estimated 419 and 73 female Ae. aegypti, respectively. No significant decrease in collection size of female Ae. aegypti could be attributed to the treatments. In the wet season, 243 SLOs and 15 BGSs killed ∼993 and 119 female Ae. aegypti, respectively. The mean number of female Ae. aegypti collected after 4 weeks with SOs and BGSs was significantly less than the control (LSD post‐hoc test). The third mass‐trapping intervention was conducted using the BLO during the wet season in Cairns. For this trial, three treatment areas were each provided with BLOs (>500, ∼4/premise) plus larval control, and an untreated control area was designated. Adult female Ae. aegypti were collected for 4 weeks pre‐ and post‐treatment using 15 BGSs and 20 SOs. During this period, 53.2% of BLOs contained a total of 6654 Ae. aegypti eggs. Over the intervention period, collections of Ae. aegypti in the treatment areas were significantly less than in the control area for BGSs but not SOs. An influx of relatively large numbers of young females may have confounded the measurement of changes in populations of older females in these studies. This is an important issue, with implications for assessing delayed action control measures, such as LOs and parasites/pathogens that aim to change mosquito age structure. Finally, the high public acceptability of SLOs and BLOs, coupled with significant impacts on female Ae. aegypti populations in two of the three interventions reported here, suggest that mass trapping with SLOs and BLOs can be an effective component of a dengue control strategy.

Bacillus thuringiensis var. israelensis (Bti) Provides Residual Control of Aedes aegypti in Small Containers

We examined the use of megadoses of VectoBac WG for residual control of Aedes aegypti in 2-L plastic buckets. Doses of 10x, 20x, and 50x the recommended rate of 8 mg/L provided >/= 90% control for 8, 8, and 23 weeks, respectively. There was no significant difference in mortality between dry (neat) or aqueous mixture of VectoBac WG. Pretreatment of dry containers up to 8 weeks before flooding did not significantly decrease efficacy through 11 success weeks. Thus, megadoses of dry formulations of Bti can be used for residual control of Ae. aegypti in small containers. Furthermore, these doses use small amounts of product (0.08-0.4 g/L) that is more practical to measure than the minute amounts (0.008 g/L) required by the recommended rate, and cost US

Rapid Estimation of Aedes aegypti Population Size Using Simulation Modeling, with a Novel Approach to Calibration and Field Validation

2.18 to treat 50 Cairns yards containing an average total of 80 containers. This method could also be used to control Aedes albopictus.

Genetic Structure of Aedes aegypti in Australia and Vietnam Revealed by Microsatellite and Exon Primed Intron Crossing Markers Suggests Feasibility of Local Control Options

Abstract New approaches for control of the dengue vector Aedes aegypti (L.) are being developed, including the potential introduction of life-shortening symbiont bacteria into field populations and the release of transgenic strains with reduced vector competency. With these new approaches comes the need for rapid estimations of existing field population size. Here, we describe the use of simulation modeling with container-inhabiting mosquito simulation (CIMSiM) for estimation of Ae. aegypti pupal crop size in north Queensland, Australia. CIMSiM was calibrated for local conditions by deploying “sentinel key containers” (tire, 2-liter plastic bucket, 0.6-liter pot plant base, and tarpaulin indentation) in which water flux and pupal productivity were studied for 72 d. Iterative adjustment of CIMSiM parameters was used to fit model outputs to match that of sentinel key containers. This calibrated model was then used in a blind field validation, in which breeding container and local meteorological data were used to populate CIMSiM, and model outputs were compared with a field pupal survey. Actual pupae per ha during two 10-d periods in 2007 fell within 95% confidence intervals of simulated pupal crop estimates made by 10 replicate simulations in CIMSiM, thus providing a successful field validation. Although the stochasticity of the field environment can never be wholly simulated, CIMSiM can provide field-validated estimates of pupal crop in a timely manner by using simple container surveys.

A lethal ovitrap-based mass trapping scheme for dengue control in Australia: I. Public acceptability and performance of lethal ovitraps

ABSTRACT The distribution of Aedes aegypti (L.) in Australia is currently restricted to northern Queensland, but it has been more extensive in the past. In this study, we evaluate the genetic structure of Ae. aegypti populations in Australia and Vietnam and consider genetic differentiation between mosquitoes from these areas and those from a population in Thailand. Six microsatellites and two exon primed intron crossing markers were used to assess isolation by distance across all populations and also within the Australian sample. Investigations of founder effects, amount of molecular variation between and within regions and comparison of FST values among Australian and Vietnamese populations were made to assess the scale of movement of Ae. aegypti. Genetic control methods are under development for mosquito vector populations including the dengue vector Ae. aegypti. The success of these control methods will depend on the population structure of the target species including population size and rates of movement among populations. Releases of modified mosquitoes could target local populations that show a high degree of isolation from surrounding populations, potentially allowing new variants to become established in one region with eventual dispersal to other regions.

Susceptibility of Eucalyptus globulus to Mnesampela privata defoliation in relation to a specific foliar wax compound

We report on the first field evaluation of the public acceptability and performance of two types of lethal ovitrap (LO) in three separate trials in Cairns, Australia. Health workers were able to set standard lethal ovitraps (SLOs) in 75 and 71% of premise yards in the wet and dry season, respectively, and biodegradable lethal ovitraps (BLOs) in 93% of yards. Public acceptance, measured as retention of traps by residents, was high for both trap types, with <9% of traps missing after 4 weeks. Traps retaining water after 4 weeks were 78 and 34% for the two SLO trials and 58% for the BLOs. The ‘failure rate’ in the 535 BLOs set in the field for 4 weeks was 47%, of which 19% were lost, 51% had holes from probable insect chewing, 23% were knocked over, 7% had dried by evaporation and 1% were split. There was no significant difference in the failure rate of BLOs set on porous (grass, soil and mulch) versus solid (tiles, concrete, wood and stone) substrates. The SLOs and the BLOs were readily acceptable to ovipositing Aedes aegypti L. (Diptera: Culicidae); the mean number of eggs/trap was 6 and 15, for the dry season and wet season SLO trial, respectively, and 15 for the BLO wet season trial. Indeed, 84–94% of premise yards had egg positive SLOs or BLOs. A high percentage of both wet and dry season SLOs (29 and 70%, respectively) and BLOs (62%) that were dry after 4 weeks were egg positive, indicating the traps had functioned. Lethal strips from SLOs and BLOs that had been exposed for 4 weeks killed 83 and 74%, respectively, of gravid Ae. aegypti in laboratory assays. These results indicate that mass trapping schemes using SLOs and BLOs are not rejected by the public and effectively target gravid Ae. aegypti. The impact of the interventions on mosquito populations is described in a companion paper.

Constitutive or induced defences - how does Eucalyptus globulus defend itself from larval feeding?

Summary. Increased abundance of several aliphatic benzyl and phenylethyl alkanoates were previously associated with reduced defoliation of Eucalyptus globulus Labill. by the autumn gum moth (Mnesampela privata Guenée) in two clonally replicated F2 families. We examine the robustness of this association by comparing the abundance of the implicated foliar wax compounds with defoliation levels in three plantation sites and a genetics trial in northern Tasmania. At all locations, the aliphatic alkanoate, benzyl n-tetracosanoate (C24), was found in significantly higher abundance in the foliar wax of E. globulus trees that had displayed low M. privata damage (designated resistant) compared to trees that had displayed high damage (designated susceptible). Further, when sites were combined, benzyl n-tetracosanoate (C24) accounted for 16 % of variation in field defoliation. Field defoliation was also positively correlated to M. privata oviposition both in the field and on foliage sprigs in a cage. In the cage bioassay, oviposition accounted for 20 % of variation in field defoliation making it a better predictor of tree defoliation in the field than either field oviposition, which accounted for 9 % of variation, or screening foliage for the abundance of benzyl n-tetracosanoate (C24). Despite both benzyl n-tetracosanoate (C24) and oviposition being related to field defoliation, there was no statistically significant relationship between the abundance of benzyl n-tetracosanoate (C24) and oviposition in the field or cage bioassay. Further work is therefore required to determine if increased levels of this compound in the foliar wax operates as a direct deterrent to M. privataoviposition.

Genetic variation in Eucalyptus globulus in relation to susceptibility from attack by the southern eucalypt leaf beetle, Chrysophtharta agricola

Summary.Following herbivory, induced responses involving plant secondary metabolites have been reported in a number of tree species. Although a wide range of plant secondary metabolites appear to operate as constitutive plant defences in trees belonging to the Eucalyptus genus, no induced responses have as yet been reported following foliar-chewing insect damage. We empirically tested whether branch defoliation (artificial and larval) of 2-year-old Eucalyptus globulus Labill. trees altered the abundance of specific plant secondary metabolites immediately (3 months after initial larval feeding) and 8 months after the cessation of larval feeding. Metabolites assayed, included essential oils, polyphenolic groups and foliar wax compounds and in all cases their abundance was not significantly altered by defoliation. However, the level of foliar tannins after 3 months of larval feeding did display a trend that suggested elevated levels as the result of defoliation, though this trend was not evident 8 months later, indicating that, if real, the response was a rapid and not a delayed induced response. The level of foliar tannins was also negatively correlated to both average larval survival and average percentage branch defoliation, suggesting that foliar tannins may operate as toxins and/or anti-feedants to M. privata larval feeding.

Field validation of a transcriptional assay for the prediction of age of uncaged Aedes aegypti mosquitoes in northern Australia.

The southern eucalypt leaf beetle, Chrysophtharta agricola (Chapuis), is an outbreak insect pest of commercial Eucalyptus globulus Labill. plantations in south-eastern Australia. We surveyed a young E. globulus family trial in southern Tasmania to determine whether genetic variation existed in the susceptibility of trees to C. agricola field oviposition. The family trial consisted of 225 families, derived from open-pollinated seed collected from native stands at 24 different localities, representing nine geographic subraces. The survey showed that E. globulus subraces from Victoria were significantly more susceptible to C. agricola oviposition than Tasmanian subraces. Significant additive genetic variation within subraces was evident for the number of egg batches, larval clutches and their combination (infestation level), although these heritability scores were all low (egg batches h2op = 0.09; larval clutches h2op = 0.14 and infestation level h2op = 0.11). Subsequent tree defoliation was significantly positively correlated with infestation at a phenotypic, genetic and environmental level. No significant differences in C. agricola oviposition on foliage sprigs was detected among subraces in an ex situ caged oviposition experiment. However, the cage experiment proved to be a good predictor of field oviposition, accounting for 70 and 88% of variation in field oviposition among localities and subraces, respectively.

Oviposition by autumn gum moth (Mnesampela privata)in relation to Eucalyptus globulus defoliation,larval performance and natural enemies

Background New strategies to eliminate dengue have been proposed that specifically target older Aedes aegypti mosquitoes, the proportion of the vector population that is potentially capable of transmitting dengue viruses. Evaluation of these strategies will require accurate and high-throughput methods of predicting mosquito age. We previously developed an age prediction assay for individual Ae. aegypti females based on the transcriptional profiles of a selection of age responsive genes. Here we conducted field testing of the method on Ae. aegypti that were entirely uncaged and free to engage in natural behavior. Methodology/Principal Findings We produced “free-range” test specimens by releasing 8007 adult Ae. aegypti inside and around an isolated homestead in north Queensland, Australia, and recapturing females at two day intervals. We applied a TaqMan probe-based assay design that enabled high-throughput quantitative RT-PCR of four transcripts from three age-responsive genes and a reference gene. An age prediction model was calibrated on mosquitoes maintained in small sentinel cages, in which 68.8% of the variance in gene transcription measures was explained by age. The model was then used to predict the ages of the free-range females. The relationship between the predicted and actual ages achieved an R2 value of 0.62 for predictions of females up to 29 days old. Transcriptional profiles and age predictions were not affected by physiological variation associated with the blood feeding/egg development cycle and we show that the age grading method could be applied to differentiate between two populations of mosquitoes having a two-fold difference in mean life expectancy. Conclusions/Significance The transcriptional profiles of age responsive genes facilitated age estimates of near-wild Ae. aegypti females. Our age prediction assay for Ae. aegypti provides a useful tool for the evaluation of mosquito control interventions against dengue where mosquito survivorship or lifespan reduction are crucial to their success. The approximate cost of the method was US