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Mosquitoes are the main arthropod vectors of human pathogens. The current methods for mosquito identification include morphological and molecular methods. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), now routinely used for bacterial identification, has recently emerged in the field of entomology. The aim of this study was to use MALDI-TOF MS to identify mosquito colonies from French Polynesia. Five hundred specimens from French Polynesia belonging to three species, Aedes aegypti, Aedes polynesiensis, and Culex quinquefasciatus, were included in the study. Testing the legs of these mosquitoes by MALDI-TOF MS revealed a 100% correct identification of all specimens at the species level. The MALDI-TOF MS profiles obtained allowed differentiation of male from female mosquitoes and the specific identification of female mosquito colonies of the same species but different geographic origin.

The circulation of arthropod-borne viruses (arboviruses) throughout the Pacific is no exception in French Polynesia (FP). We review here the mosquito vectors involved or potentially involved in the transmission of arboviruses in FP. We highlight Aedes aegypti and Aedes polynesiensis; species that are widely distributed in FP and whose ability to transmit dengue, zika and chikungunya viruses is well known. In addition, we discuss the contribution of those species and the abundant Culex annulirostris to the silent circulation of Ross River virus in FP. As these mosquito species and others present in FP are known to be vectors of a large panel of arboviruses, there is a high risk for new emergences. Keywords: Aedes aegypti, Aedes polynesiensis, arboviruses, chikungunya virus, Culex annulirostris, dengue virus, French Polynesia, mosquito vectors, Ross River virus, Zika virus

Vector control is critical to limit the circulation of vector-borne diseases, like chikungunya, dengue or zika, which have become important issues around the world. Among them, the Sterile Insect Technique (SIT) and the Incompatible Insect Technique (IIT) have recently aroused a renewed interest. In this paper we derive and study a minimalistic mathematical model designed for Aedes mosquito population elimination by SIT/IIT. Contrary to most of the previous models, it is bistable in general, allowing simultaneously for elimination of the population and for its survival. We consider different types of releases (constant, periodic or impulsive) and show necessary conditions to reach elimination in each case. We also estimate both sufficient and minimal treatment times. Biological parameters are estimated from a case study of an Aedes polynesiensis population, for which extensive numerical investigations illustrate the analytical results. The applications of this work are two-fold: to help identifying some key parameters that may need further field investigations, and to help designing release protocols.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://parasitesandvectors.biomedcentral.com/articles/10.1186/s13071-018-3221-x Background: Several mosquito population suppression strategies based on the rearing and release of sterile males have provided promising results. However, the lack of an efficient male selection method has hampered the expansion of these approaches into large-scale operational programmes. Currently, most of these programmes targeting Aedes mosquitoes rely on sorting methods based on the sexual size dimorphism (SSD) at the pupal stage. The currently available sorting methods have not been developed based on biometric analysis, and there is therefore potential for improvement. We applied an automated pupal size estimator developed by Grupo Tragsa with laboratory samples of Anopheles arabiensis, Aedes albopictus, Ae. polynesiensis, and three strains of Ae. aegypti. The frequency distribution of the pupal size was analyzed. We propose a general model for the analysis of the frequency distribution of mosquito pupae in the context of SSD-sorting methods, which is based on a Gaussian mixture distribution functions, thus making possible the analysis of performance (% males recovery) and purity (% males on the sorted sample). Results: For the three Aedes species, the distribution of the pupae size can be modeled by a mixture of two Gaussian distribution functions and the proposed model fitted the experimental data. For a given population, each size threshold is linked to a specific outcome of male recovery. Two dimensionless parameters that measure the suitability for SSD-based sorting of a specific batch of pupae are provided. The optimal sorting results are predicted for the highest values of SSD and lowest values of intra-batch variance. Rearing conditions have a strong influence in the performance of the SSD-sorting methods and non-standard rearing can lead to increase pupae size heterogeneity. Conclusions: Sex sorting of pupae based on size dimorphism can be achieved with a high performance (% males recovery) and a reasonably high purity (% males on the sorted sample) for the different Aedes species and strains. The purity and performance of a sex sorting operation in the tested Aedes species are linked parameters whose relation can be modeled. The conclusions of this analysis are applicable to all the existing SSD-sorting methods. The efficiency of the SSDsorting methods can be improved by reducing the heterogeneity of pupae size within rearing containers. The heterogeneity between batches does not strongly affect the quality of the sex sorting, as long as a specific separation threshold is not pre-set before the sorting process. For new developments, we recommend using adaptive and precise threshold selection methods applied individually to each batch or to a mix of batches. Adaptive and precise thresholds will allow the sex-sorting of mixed batches in operational conditions maintaining the target purity at the cost of a reduction in performance. We also recommend a strategy whereby an acceptable level of purity is pre-selected and remains constant across the different batches of pupae while the performance varies from batch to batch to fit with the desired purity.

The impact of deltamethrin-impregnated cloth targets on Stegomyia polynesiensis (= Aedes polynesiensis) (Marks) (Diptera: Culicidae) was assessed under laboratory and semi-field settings in French Polynesia. Stegomyia polynesiensis females were released into small laboratory cages and large field cages containing either a deltamethrin-treated or an untreated navy blue cloth, and mosquito knock-down and mortality were assessed. The 24-h mortality rate in mosquitoes exposed to the insecticide-treated target in small cages was 98.0%. These mosquitoes also demonstrated significantly higher levels of knock-down than those exposed to the untreated target. Mortality in field cages was assessed at 24 and 48 h. The 24-h mortality rate in mosquitoes exposed to the control target was 31.2%, whereas that in those exposed to the deltamethrin-treated target was 54.3%. The 48-h mortality rate was also elevated in mosquitoes exposed to the deltamethrin-treated target, but this result did not differ significantly from that observed in mosquitoes exposed to the control target. The significant suppression of female S. polynesiensis by deltamethrin-treated resting targets in this study indicates that these targets could play a role in the control of an important disease vector in the South Pacific region.

Background Aedes mosquitoes severely affect the health and wellbeing of human populations by transmitting infectious diseases. In French Polynesia, Aedes aegypti is the main vector of dengue, chikungunya and Zika, and Aedes polynesiensis the primary vector of Bancroftian filariasis and a secondary vector of arboviruses. Tools for assessing the risk of disease transmission or for measuring the efficacy of vector control programmes are scarce. A promising approach to quantify the human-vector contact relies on the detection and the quantification of antibodies directed against mosquito salivary proteins. Methodology/Principal findings An ELISA test was developed to detect and quantify the presence of immunoglobulin G (IgG) directed against proteins from salivary gland extracts (SGE) of Ae. aegypti and Ae. polynesiensis in human populations exposed to either species, through a cross-sectional study. In Tahiti and Moorea islands where Ae. aegypti and Ae. polynesiensis are present, the test revealed that 98% and 68% of individuals have developed IgG directed against Ae. aegypti and Ae. polynesiensis SGE, respectively. By comparison, ELISA tests conducted on a cohort of people from metropolitan France, not exposed to these Aedes mosquitoes, indicated that 97% of individuals had no IgG directed against SGE of either mosquito species. The analysis of additional cohorts representing different entomological Aedes contexts showed no ELISA IgG cross-reactivity between Ae. aegypti and Ae. polynesiensis SGE. Conclusions/Significance The IgG response to salivary gland extracts seems to be a valid and specific biomarker of human exposure to the bites of Ae. aegypti and Ae. polynesiensis. This new immuno-epidemiological tool will enhance our understanding of people exposure to mosquito bites, facilitate the identification of areas where disease transmission risk is high and permit to evaluate the efficacy of novel vector control strategies in Pacific islands and other tropical settings., Author summary In Pacific islands like in most tropical regions, Aedes mosquitoes affect the health of human populations by transmitting diseases like dengue, chikungunya, Zika and filariasis. The biting nuisance of Aedes mosquitoes also impacts local tourism, affecting the sustainability of island economies. Mosquito saliva is injected during the biting process, and the response triggered by the human immune system to proteins contained in mosquito saliva was shown to be a relevant biomarker of exposure to mosquito bites. Using this approach, we have developed an immuno-epidemiological tool to investigate the exposure of people to the bites of Aedes aegypti and Aedes polynesiensis, two significant mosquito vectors of infectious diseases in French Polynesia and other island countries and territories in the Pacific. This novel tool proved specific and reliable. It will improve the assessment of disease transmission risk and be useful for measuring the efficacy of both conventional and innovative vector control strategies.

SummaryTo control mosquito populations for managing vector-borne diseases, a critical need is to identify and predict their response to causal environmental variables. However, most existing attempts rely on linear approaches based on correlation, which cannot apply in complex, nonlinear natural systems, because correlation is neither a necessary nor sufficient condition for causation. Appling empirical dynamic modelling that acknowledges nonlinear dynamics on nine subpopulations of tiger mosquitos from three neighbouring reef islets of the Raiatea atoll, we identified temperature, precipitation, dew point, air pressure, and mean tide level as causal environmental variables. Interestingly, responses of subpopulations in close proximity (100-500 m) differed with respect to their causal environmental variables and the time delay of effect, highlighting complexity in mosquito-environment causality network. Moreover, we demonstrated how to explore the effects of changing environmental variables on number and strength of mosquito outbreaks, providing a new framework for pest control and disease vector ecology.

The Gravid Aedes Trap (GAT) is a passive trap that relies on visual and olfactory cues to lure and capture gravid mosquitoes. The GAT was designed as a dengue vector surveillance tool for use in conditions where power sources are not readily available. Experiments were conducted over a 10-d period in Fiji to determine which species of mosquitoes the GAT would collect, how different infusion types affect the attractiveness of the trap, how long each infusion type took to begin attracting mosquitoes, and how long the infusion was attractive to container breeding Aedes mosquitoes. Infusions were created by adding 10 g of organic material to 2 liters of water. Infusions were made using chicken feed pellets, dried mango leaves (Mangifera indica), and dried lawn grasses (primarily Axonopus spp.). The GAT collected four different vectors Aedes aegypti (Linnaeus) (Diptera: Culicidae), Aedes albopictus (Skuse) (Diptera: Culicidae), Aedes polynesiensis Mark (Diptera: Culicidae), and Culex quinquefasciatus Say (Diptera: Culicidae). As observed in previous studies, using some type of organic infusion improved the attractiveness of the trap. Of the three different infusion components tested, chicken feed was most attractive followed by lawn grasses and mango leaves. All infusions performed better than plain water. Chicken feed was found to be most attractive for Ae. aegypti and Ae. albopictus during the first 6 d after placing the traps and for Cx. quinquifasciatus in days 7-9. The mango and grass infusions took longer to attract mosquitoes but were most attractive during days 7-10 for all species.

Aedes polynesiensis Marks (Diptera: Culicidae) larvae were reared to adulthood in the laboratory under a range of temperatures and larval densities. We studied the effect of these variables on several life table parameters of relevance to male-release-based vector control strategies including: larval survivorship, developmental time to pupation, male to female ratio, male pupae yield, adult male size and survival. The range of tested rearing temperatures (20, 25, 27, and 30 °C) and larval densities (50, 100, 200, and 400 larvae/L) was selected within the conditions allowing larval growth and survival. Larval survivorship was the highest when larvae were reared at 200 larvae/L for all temperatures except 20 °C. Male to female ratio was male biased at all temperatures and densities. Time to pupation decreased with increasing temperatures. Larval density and temperature influenced the proportion of males pupating on first day of pupation with 43-47% of total male pupae produced at 25 °C. No significant differences in mean wing length were observed between male mosquitoes reared in the laboratory (except at 20 and 30 °C for some densities) and field collected males. Altogether, the study allowed the identification of rearing conditions delivering high male yield with essentially no female contamination, adequate adult male size and survival. Ae. polynesiensis thus appears particularly amenable to biological and mechanical sex separation offering good prospects for Ae. polynesiensis population suppression trials that rely on the production and release of large numbers of incompatible or sterile males.

Understanding the mating competitiveness of male mosquitoes in field settings is essential to programs relying on the mass release of modified male mosquitoes, yet studies on male ecology have been hampered by the lack of a convenient trapping method. An existing promising method makes use of the innate attraction of males to female flight tones. Here, we present laboratory, greenhouse, and field experiments on the efficacy of sound traps for the collection of Aedes aegypti (L.) and Aedes polynesiensis Marks, and laboratory experiments with Ae. aegypti on the effects of male age, size, and mating status on responsiveness to a range of frequencies. Age and mating status influenced the overall responsiveness to sound, whereas male size did not. There were no interactions between these factors and sound frequency. A Centers for Disease Control and Prevention miniature light trap modified to produce a tone of 465 Hz collected 76.2% of Ae. aegypti males in laboratory cages, and 49.7% of males in a greenhouse enclosure. In two sets of experiments in laboratory cages, 50.8 and 46.5% of male Ae. polynesiensis were captured with a trap producing a tone of 440 Hz. In the field, CDC miniature light traps or BG-Sentinel traps fitted with a portable speaker producing tones of 440 or 465 Hz captured significantly more male Ae. polynesiensis when placed near a male swarm than did traps that did not produce sound. When the trap was placed at a distance of 16.5 m from the nearest swarm, there was no significant difference in the number of males caught between control and sound-producing traps. The numbers of Ae. aegypti males captured were low under all circumstances in the field.

We characterize the swarming behavior of male Aedes polynesiensis (Marks) in American Samoa. Instead of swarming around a blood host, males used the base of certain trees as a marker. Repeated sampling proved nondestructive and allowed us to investigate the impact of static (e.g., tree species) and dynamic (e.g., barometric pressure) characters on the likelihood of swarm presence and intensity. Tree circumference and oviposition activity (number of Ae. polynesiensis reared from oviposition cups) were significant positive predictors of the number of males in a swarm. Tree circumference and diameter were significantly positively associated, and canopy height was significantly negatively associated, with swarm occurrence. Comparisons between males swarming early and late during the swarming period allowed for insight into swarm composition in terms of male size and the amount of putative fluid (e.g., nectar) in the crop, indicators of energetic reserves. Males collected during the late period had significantly larger wings and less crop contents than did males of the early cohort. Because the ecology of male Ae. polynesiensis remains understudied, we consider how the current results could facilitate further studies related to applied autocidal strategies as well as the evolution of host-based mating behavior.

Aedes polynesiensis Marks (Diptera: Culicidae) is the primary vector of lymphatic filariasis (LF) in the island countries and territories of the South Pacific. In the development of a novel control tool, the response of Ae. polynesiensis to six different colours (three solid fabrics, two patterned fabrics and a plastic tarp) was measured using a digital photographic system. Adult mosquitoes were placed into an environmental chamber and allowed to choose between a white target and one of six experimental targets. Mosquito landing frequency and landing duration were calculated. Adult female Ae. polynesiensis preferred all of the experimental targets to the white control target. Mosquito landing frequency was highest for the solid targets (black, navy blue and red) followed in turn by the two colour pattern targets and the polyethylene target. Mosquito landing duration was greater for experimental targets when compared with white control targets. Mosquito landing frequencies did not change over time during the course of the assay. The response of male Ae. polynesiensis was also measured when exposed to a 100% cotton black target. Male mosquitoes preferred the black target to the white control target, although at levels lower than that observed in female mosquitoes. The results suggest that future investigations evaluating the visual responses of Ae. polynesiensis mosquitoes are warranted, with a special emphasis on semi-field and field-based experiments.

Background In 2013–2014, French Polynesia experienced for the first time a Zika outbreak. Two Aedes mosquitoes may have contributed to Zika virus (ZIKV) transmission in French Polynesia: the worldwide distributed Ae. aegypti and the Polynesian islands-endemic Ae. polynesiensis mosquito. Methodology/Principal Findings To evaluate their vector competence for ZIKV, mosquitoes were infected per os at viral titers of 7 logs tissue culture infectious dose 50%. At several days post-infection (dpi), saliva was collected from each mosquito and inoculated onto C6/36 mosquito cells to check for the presence of ZIKV infectious particles. Legs and body of each mosquito were also collected and submitted separately to RNA extraction and ZIKV RT-PCR. In Ae. aegypti the infection rate was high as early as 6 dpi and the dissemination efficiency get substantial from 9 dpi while the both rates remained quite low in Ae. polynesiensis. The transmission efficiency was poor in Ae. aegypti until 14 dpi and no infectious saliva was found in Ae. polynesiensis at the time points studied. Conclusions/Significance In our experimental conditions, the late ability of the French Polynesian Ae. aegypti to transmit ZIKV added by the poor competence of Ae. polynesiensis for this virus suggest the possible contribution of another vector for the propagation of ZIKV during the outbreak, in particular in remote islands where Ae. polynesiensis is predominating., Author Summary From 2007, Zika virus has caused several outbreaks in the Pacific including French Polynesia. Aedes aegypti mosquito which is present in almost all Pacific Island Countries is reasonably expected to have been involved in the Zika outbreaks. In addition endemic Aedes mosquito species may have sustained Zika virus transmission in the less urbanized and most remote islands. In the present study we provide for the first time data about the vector competence of the endemic Ae. polynesiensis species for Zika virus. We found under experimental conditions a weak competence of Ae. polynesiensis for the virus. Furthermore we demonstrated a late ability of the French Polynesian population of Ae. aegypti to transmit Zika virus. These findings raise questions about the potential involvement of other vector(s) in Zika virus transmission in place or together with the Aedes mosquitoes. In a context where innovative vector control strategies are mostly focused on targeting the mosquito species considered as the main arbovirus vectors, the potential for others vector species to take the lead in transmitting such arboviruses should not be neglected.

The efficacy of the BG-Sentinel (BGS) and the BG-Mosquitito (BGM) mosquito traps for sampling populations of the important filariasis and dengue vector Aedes (Stegomyia) polynesiensis (Marks) was evaluated in French Polynesia against human bait collections (HBC) using a modified Centers for Disease Control and Prevention backpack aspirator. Traps were baited with BG-Lure (a combination of lactic acid, ammonia, and caproic acid) or carbon dioxide plus octenol (1-octen-3-ol) known as attractants to aedine mosquitoes. Mosquito sampling was conducted on two typical islands of French Polynesia: the high, volcanic island of Moorea, and the low, coral island (atoll) of Tetiaroa Sampling efficacy was measured in a randomized Latin Square design. Production of carbon dioxide from yeast-sugar fermentation was used as an alternative source of CO2 because supply via dry ice, gas cylinders, or propane combustion in remote tropical islands is costly and challenging. Although the BGS trap captured the greatest number ofAe. polynesiensis in both island settings, catch rates of BGS or BGM baited with either lure were not significantly different from that of HBC. On Moorea, the number of collected aedes species in the BGS trap baited with either lure was significantly greater than the BGM with BG-lure. On Tetiaroa, BGM trapping was severely hampered by damage from rats, and the traps were removed from the study. Our study confirms the efficiency, comparability, and convenience of the BGS trap, a robust and safe alternative to HBC for sampling Aedes mosquitoes in research and surveillance efforts against filariasis and arboviruses in the South Pacific.

Mark-release-recapture methods were used to compare Aedes polynesiensis Marks adult numbers and dispersal between dry and wet seasons in a closed population on a small island (motu) in French Polynesia. Females were more than three times more common during wet (December 2008) than dry (May 2007) season samplings although high numbers of vectors were collected during both seasons. Lincoln—Petersen estimates for Ae. polynesiensis females on the motu were 6,055 per hectare for the dry season and 18,860 per hectare for the wet season. Marked females dispersed rapidly to all parts of the motu and survived until recaptures on days 1–5 after release. Males were not adequately sampled using human sentinels or Biogent Sentinel traps.

Background From October 2014 to March 2015, French Polynesia experienced for the first time a chikungunya outbreak. Two Aedes mosquitoes may have contributed to chikungunya virus (CHIKV) transmission in French Polynesia: the worldwide distributed Ae. aegypti and the Polynesian islands-endemic Ae. polynesiensis mosquito. Methods To investigate the vector competence of French Polynesian populations of Ae. aegypti and Ae. polynesiensis for CHIKV, mosquitoes were exposed per os at viral titers of 7 logs tissue culture infectious dose 50%. At 2, 6, 9, 14 and 21 days post-infection (dpi), saliva was collected from each mosquito and inoculated onto C6/36 mosquito cells to check for the presence of CHIKV infectious particles. Legs and body (thorax and abdomen) of each mosquito were also collected at the different dpi and submitted separately to viral RNA extraction and CHIKV real-time RT-PCR. Results CHIKV infection rate, dissemination and transmission efficiencies ranged from 7–90%, 18–78% and 5–53% respectively for Ae. aegypti and from 39–41%, 3–17% and 0–14% respectively for Ae. polynesiensis, depending on the dpi. Infectious saliva was found as early as 2 dpi for Ae. aegypti and from 6 dpi for Ae. polynesiensis. Our laboratory results confirm that the French Polynesian population of Ae. aegypti is highly competent for CHIKV and they provide clear evidence for Ae. polynesiensis to act as an efficient CHIKV vector. Conclusion As supported by our findings, the presence of two CHIKV competent vectors in French Polynesia certainly contributed to enabling this virus to quickly disseminate from the urban/peri-urban areas colonized by Ae. aegypti to the most remote atolls where Ae. polynesiensis is predominating. Ae. polynesiensis was probably involved in the recent chikungunya outbreaks in Samoa and the Cook Islands. Moreover, this vector may contribute to the risk for CHIKV to emerge in other Polynesian islands like Fiji, and more particularly Wallis where there is no Ae. aegypti., Author Summary Chikungunya virus has caused a series of outbreaks in the Pacific from 2011, including French Polynesia. Aedes (Ae.) aegypti mosquito, which has colonized almost all Pacific Island Countries, is reasonably expected to have been involved in the chikungunya outbreaks. In addition, endemic Aedes mosquito species may have sustained chikungunya virus transmission in the less urbanized and most remote islands. In the present study, we demonstrated the ability of French Polynesian populations of Ae. aegypti and Ae. polynesiensis to replicate, disseminate and transmit chikungunya virus under experimental conditions. Our results provide for the first time clear evidence for Ae. polynesiensis to act as an efficient chikungunya vector. These findings corroborate previous observation that endemic Aedes species, like Ae. hensilli in Yap Island, may play a critical role in sustaining chikungunya virus transmission, in place or together with the widely distributed Ae. aegypti and Ae. albopictus. In a context where innovative vector control strategies are mostly focused on targeting the mosquito species considered as the main arbovirus vectors, the potential for endemic Aedes species to take the lead in transmitting such arboviruses should not be neglected.

The endosymbiotic bacteria Wolbachia manipulate host reproduction by inducing a form of sterility known as cytoplasmic incompatibility (CI), promoting the invasion of infection into natural host populations. CI has received attention for use in applied strategies to control insect vectors of disease. Thus, to understand both naturally occurring Wolbachia invasions and evaluate potential applied strategies, it is important to understand Wolbachia interactions with its host, including impacts on fitness and the CI level. In this study, we examined for an effect of Wolbachia on survivorship, developmental time, sex ratio, longevity, fecundity, and egg hatch of Aedes polynesiensis Marks, which is the primary vector of Wuchereria bancrofti in the South Pacific. In this study, we have compared strains of A. polynesiensis that are naturally and artificially infected with Wolbachia and additional strains that are aposymbiotic (Wolbachia removed to generate an uninfected strain). Artificially infected strains were observed to have increased larval mortality and decreased adult longevity when compared with aposymbiotic strains. Naturally infected strains were observed to have decreased larval mortality, pupal mortality, increased adult longevity, and a larger adult size when compared with aposymbiotic strains. Artificially infected males that were 4 wk old were able to induce high rates of CI, similar to young males. We discuss the results in relation to the natural spread of Wolbachia and Wolbachia-based applied strategies to modify A. polynesiensis populations.

Successful elimination of lymphatic filariasis (LF) requires accurate identification of residual foci of transmission and stringent surveillance strategies to combat potential resurgence. This is challenging in areas where the day-biting Aedes polynesiensis is endemic, such as Samoa, since in previous studies no geographical clustering of infection has been demonstrated. Another challenge for this low prevalence phase is the choice of diagnostic assay as testing for circulating filarial antigen (CFA) or microfilariae (Mf) alone may not have adequate sensitivity. This could be solved by using the commercially available filariasis Cellabs enzyme linked immunosorbent assay (CELISA) to measure antibody. In the current study five Samoan villages were chosen based on previous epidemiological assessments to represent a range of infection prevalences. CFA, Mf, and antibody levels in children ≤ 10 years had been recorded and results linked to household of residence and/or primary school of attendance. To ascertain the location of exposure, two scenarios based on potential foci of transmission around communities and schools were explored. Both scenarios revealed significant spatial clusters of households with infected individuals and a relationship to antibody positive children when they were included in the spatial analysis. Fasitoo-Tai had the highest LF prevalence and largest geographical spatial clusters for both scenarios. In Falefa, spatial clusters were detected only for the primary school scenario. In Tafua, which spanned an area of 19.5 km(2), no spatial clusters were detected. Lastly, in Siufaga, the village with the lowest LF prevalence, significant clustering of infected individuals was observed and, for the primary school scenario, this was geographically related to exposure. These promising findings are the first published evidence of spatial clustering of LF in a day-biting Ae. polynesiensis endemic area.

Larval and pupal surveys of 439 natural and 2,455 domestic containers (total of 2,894 containers) were undertaken in four villages in American Samoa during the wet and dry seasons. For the first time, larvae and pupae of Ae. oceanicus were found in a variety of domestic containers (including buckets, plastic and polystyrene containers, cans, and tires) in addition to their traditionally reported habitats of plant leaf axils. Finding Ae. oceanicus in artificial containers in three villages during both the wet and dry seasons suggests that Ae. oceanicus is adapting to use these increasingly abundant water sources for breeding sites. The larger water volumes held by such containers could ensure the survival of this species during prolonged dry weather periods.

The efficacy of the recently developed BG-Sentinel mosquito trap baited with BG-Lure (a combination of lactic acid, ammonia, and caproic acid) was evaluated in American Samoa against the omnidirectional Fay-Prince trap and the Centers for Disease Control and Prevention (CDC) light trap, both baited with carbon dioxide. The BG-Sentinel trap captured the greatest number of the important filariasis and dengue vector Aedes (Stegomyia) polynesiensis at all 3 collection locations; however, its catch rate was not significantly different from that of the Fay-Prince trap at 2 of the 3 trapping locations. The CDC light trap caught very few Ae. polynesiensis. The Fay-Prince trap was more efficient than the other 2 traps for collecting Aedes (Aedimorphus) nocturnus, Aedes (Finlaya) spp., Culex quinquefasciatus, and Culex annulirostris. The efficacy and convenience of the BG-Sentinel suggest further research is warranted to evaluate its potential as a possible efficient and safe alternative to landing catches for sampling Ae. polynesiensis in research and control efforts against filariasis and dengue in the South Pacific.

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

Two surveys of Rarotonga, Cook Islands (21°20'S, 160°16'W) were made to determine the mosquito fauna of the island, and to identify the habitats required for breeding by searching for larvae. The first survey was made during the “dry season” in May 2001, the second during the “wet season” in February 2002. The mosquito fauna comprised four species Culex (Culex) quinquefasciatus Say, Culex (Culex) annulirostris Skuse, Aedes (Stego‐myia) aegypti (Linnaeus) and Aedes (Stegomyia) polynesiensis Marks. Larvae of the Culex species were most often found in larger natural and artificial water bodies. The Aedes species bred in both natural and artificial containers of all sizes. Ae. polynesiensis was the most widespread species, using natural holes in all regions as well as artificial containers in the urban areas. Most larvae of Ae. aegypti were located in small artificial containers. The two Aedes species are the vectors of dengue fever on the island. Mosquito control during outbreaks should specifically...

Populations of Aedes aegypti (L.) and Aedes polynesiensis (Marks) on Moorea Island, French Polynesia, the local vectors of dengue and filariasis, respectively, were sampled by landing/biting collection at nine localities on the east, north, and west coasts, during the late dry season, early and late wet season (September-May) 2003 and 2004, to investigate epidemiologically important features of the populations and compare them between regions and months. Biting rates of both species tended to increase (but not always significantly) for each species in each region each month from the late dry season to the late wet season, and the north coast region had significantly higher biting rates of Ae. aegypti. Insemination rates of Ae. polynesiensis females (94.5-98.8%) were consistently greater than those of Ae. aegypti (87.5-93.5%) throughout the study, but there was no significant difference in the insemination rates of either species between months and regions. Parity rates were generally highest in north coast samples and lowest in west coast samples for both species and generally increased (albeit not always significantly) by month for each species, with a range of 52.9-88.8% for Ae. polynesiensis and 28.6-53.6% for Ae. aegypti, although the high gravid rate (15.8-45.9%) of Ae. aegypti samples (reflecting its feeding more than once in a gonotrophic cycle) confounded both intraspecific and interspecific comparisons. Filarial infection was rare in Ae. aegypti, although both W. bancrofti and D. immitis were recorded, and infection rates in Ae. polynesiensis increased through the study period in each region for both filarias, with up to 4.6% infected and 1.4% infective for W. bancrofti and up to 6.3% infected and 2.5% infective for D. immitis. For W. bancrofti, infection rates were significantly lower on the west coast and also in the dry season, whereas rates for infective stages were significantly greater in the late wet season. For D. immitis there was no significant difference in infection rates between regions, but rates were significantly greater in the late wet season. Rainfall in all months sustains populations of both vectors and explains the relatively few significant differences between seasons; however, the wet season may provide for increased vector abundance and longevity, and present a potentially increased risk for transmission. Although the differences shown between regions also were limited in a statistical sense, there were increased risks for the northern and eastern regions, where both locals and tourists are concentrated and where the seaports and airport are located, and these areas should be priority targets for disease surveillance and vector control.

In most of the 130 islands of French Polynesia, the stenotopic mosquitoes Aedes aegypti (the main local vector for the viruses causing dengue) and Aedes polynesiensis (the main local vector of Wuchereria bancrofti) share many breeding sites in water containers such as discarded cans, coconut shells, buckets and water-storage pots and drums. In addition to selective application of insecticides, non-polluting methods of controlling these mosquitoes have been evaluated during the last decade in two main ecological situations: (1) villages, where Aedes breeding sites are typically peridomestic; and (2) flooded burrows of land crabs, the major source of Ae. polynesiensis throughout the South Pacific region. Large-scale trials of biological control agents, such as mosquito fish (Gambusia affinis and Poecilia reticulata) and copepods (Mesocyclops aspericornis), and of integrated-control strategies have demonstrated the efficacy of certain techniques and control agents against the target Aedes populations in some village situations. Generally, mechanical methods (the use of layers of polystyrene beads against mosquito larvae and pupae, and screening against adult mosquitoes) were more efficient than use of the biological control agents. By integrating several methods of control, mosquito densities (as measured by human-bait collections and larval surveys) were reduced significantly compared with the results of concurrent sampling from untreated villages, and control remained effective for months after the interventions ceased. In land-crab burrows, the first attempts to control Aedes larvae used bacterial agents (Bacillus thuringiensis) and predatory copepods gave disappointing results. Mesocyclops aspericornis could be an effective control agent if the burrows were constantly flooded, but most burrows dry out and refill periodically, so copepod populations do not survive. As it proved difficult to reach all corners of the long sinuous burrows with any control agent, larvicidal (chlorpyrifos-methyl) baits were developed for foraging crabs to carry into their burrows. This novel technique proved to be effective and could become the method of choice for treating crab burrows. Further research is underway to find the optimum (biological or chemical) larvicidal ingredient for the crab bait. Despite the ecological and logistical challenges of controlling the Aedes vectors of the dengue viruses and W. bancrofti in so many scattered islands, the French Polynesian experience indicates that relatively simple methods can be integrated and applied effectively and economically. Operationally, however, success also depends on a strong political commitment and on at-risk communities that are sufficiently motivated to maintain a good level of Aedes control.

Land crab burrows are larval mosquito habitats of major significance in the Pacific region. They are constituted by a sinuous tunnel leading to a chamber in contact with the water table, where mosquito larvae proliferate. Controlling larvae in these sites is difficult, because the configuration of burrows prevents the use of standard techniques. An experiment was carried out in French Polynesia to control Aedes polynesiensis Marks and Culex spp. breeding in burrows of the land crab Cardisoma carnifex (Herbst). The technique was based on the crab's behavior, which involves the crab carrying food into its burrow. It was shown that appetizing baits impregnated with an insecticide were carried by crabs into the flooded chamber of their burrows. A field treatment of burrows was carried out by sowing insecticide impregnated baits on the ground. The treatment coverage was almost perfect and the easy implementation of the technique enabled large areas to be treated in a short time. The bait was developed by compacting various flours, which easily incorporate a large variety of insecticide formulations. Although the baits can be easily stocked, a reliable insecticide is still to be found. The results indicate that our technique could be a method of choice for treating crab burrows.

The efficiency of the recently developed handheld InsectaZooka™ (IZ) aspirator was compared to that of the Centers for Disease Control and Prevention-Backpack (CDC-BP) aspirator by conducting human bait collections on 2 islets (locally called motus) of the atoll of Tetiaroa, French Polynesia. Abundance of mosquitoes was compared between the wind-exposed and wind-protected sides of each motu to measure the effect of wind on mosquito distribution. The number of host-seeking Aedes polynesiensis mosquitoes collected on the 2 motus with either sampling device was not significantly different. Collection of male mosquitoes was low irrespective of the type of aspirator used. Wind had an effect on mosquito distribution, as females were more abundant on the protected sides of both motus. The IZ aspirator is a lighter and equally efficient alternative to the CDC-BP aspirator for collecting Ae. polynesiensis.

Temperature effects on development of the human filarial parasite Wuchereria bancrofti (Cobbold) (Filaridea: Onchocercidae) in the main Pacific vector Aedes polynesiensis Marks (Diptera: Culicidae) are analysed in relation to ambient climatic conditions. A statistical model of the extrinsic cycle duration as a function of temperature is described and used to distinguish three patterns of W. bancrofti transmission dynamics: continuous, fluctuating and discontinuous, occurring from north to south geographically among French Polynesian archipelagos. In the northerly Marquesas Islands (8-11 degrees S) filariasis transmission is continuous and very active, facilitated by perennially high temperatures combined with constantly high rates of man-vector contact. In the southerly Australes Islands (21-28 degrees S) filariasis transmission is seasonally discontinuous and, during the cooler months (May-September), the model predicts virtually no transmission because the cycle duration exceeds the life expectancy of the vector. In the Society Islands (16-18 degrees S), between the Marquesas and Australes, transmission is predicted to be intermediate as expected from their latitude, with seasonally fluctuating transmission potential. In the Tuamotu Islands (also geographically intermediate: 14-23 degrees S), with theoretically perennial transmission potential, transmission occurs only intermittently, being limited by other human and environmental factors whereby man-vector contact is confined to seasonal agricultural situations. Generally, among French Polynesian archipelagos where Aedes polynesiensis is the vector, the transmission potential for W. bancrofti and resulting disease manifestations of lymphatic filariasis in humans are correlated with ambient temperature due to the degree of southern latitude.

Semi-annual mass DEC chemotherapy combined with vector control at the beginning of the programme, has been administered on the remote island of Maupiti (French Polynesia) since 1955 (except two periods in 1960-67 and 1970-74). The results of two surveys in 1985 and 1989, reporting 0% microfilaraemia, led to the hope that the eradication of lymphatic filariasis had been achieved. We combined parasitological criteria (microfilaraemia by membrane filtration), immunological (antigenaemia and serum levels of specific IgG antibodies) and molecular (PCR-based evaluation of infection in mosquitoes) techniques and found only good control of the parasite: We found residual microfilaraemia in 0.4% of the sample (mean level in carriers: 101.2 mf/ml), antigenaemia in 4.6% (mean level in positive persons: 714.4 units/ml) and specific IgG in 21.6% (including in one very young child). In addition, an infection rate of 1.4% was calculated in the Aedes polynesiensis vector population. These data, obtained in 1997 just before a hurricane, were partially confirmed in 1999 (0.1% of infection rate in the vector). Together with the possibility of some resistance to DEC, various epidemiological factors critical for the eradication of lymphatic filariasis are discussed.

A mark-release-recapture study was conducted to estimate the adult population size, migration, and dispersal patterns of male and female Aedes (Stegomyia) polynesiensis (Marks) in a valley of Moorea, a volcanic island, and a motu (islet) on the atoll of Tetiaroa, two settings typical of the Society Islands. Aedes polynesiensis recapture rate was high for females and low for males. The distribution of Aedes species in the valley was heterogeneous. Marked individuals dispersed to most parts of the motu and over great distances in the valley for some females. The study provides insights into the field dynamics of Ae. polynesiensis populations and confirms that more efficient sampling methods are warranted. There was no evidence of active migration between motus on the atoll, suggesting that Tetiaroa is a suitable site for small-scale initial open releases of Wolbachia incompatible insect technique and other sterile insect technique-like suppression or replacement strategies.

Attempts were made to use pteridine fluorescence levels as a tool for aging individual Aedes polynesiensis Marks and Culex quinquefasciatus Say mosquitoes. Fluorescent pigments were extracted and quantified with techniques and apparatus (i.e., a spectrofluorometer) that can be used in developing countries or in field laboratories, and that has already given consistent results in similar studies on other insect groups. However, for Ae. polynesiensis as well as Cx. quinquefasciatus, individual fluorescence measurements were mostly below the spectrofluorometer white noise level. With batches of Aedes or Culex of the same age, significant fluorescence levels were recorded but not in relation to their calendar ages. The low content of pteridines in mosquitoes suggests that standard spectrofluorometry is not sufficiently sensitive for such studies.

During 1996-1998 60,619 mosquitoes were collected around Cairns, Australia and processed for Alphavirus isolation. Thirty-three isolates of Ross River (RR) virus were made from 9 species, Aedes imprimens, Aedes kochi, Aedes notoscriptus, Aedes vigilax, Culex annulirostris, Culex gelidus, Mansonia septempunctata, Verrallina (formerly Aedes) carmenti, and Verrallina lineatus. Attempts to isolate RR virus from 121 Aedes aegypti were unsuccessful. Twenty-six (79%) of the isolates came from within 1 km of a colony of spectacled flying-foxes, Pteropus conspicillatus. The minimum infection rate for these mosquitoes was 1.0 compared with 0.2 per 1,000 for mosquitoes trapped at all other sites. Ross River virus has not previously been isolated from Ae. imprimens, Cx. gelidus, Ma. septempunctata, Ve. carmenti, or Ve. lineatus. This is also the first isolation of an arbovirus from Cx. gelidus in Australia. In conclusion, the vector status of Ve. carmenti, Ae. aegypti and Ma. septempunctata warrants further study. This study also provides evidence that P. conspicillatus may be a reservoir host.

Replicated cohorts of a Tahitian laboratory strain of Aedes polynesiensis Marks were reared at 3 larval densities with a fixed amount of food. For larvae provided with limiting per capita food ( (i.e., 400 larvae per pan with 500 mg liver powder) relative to standard rearing conditions (i.e., 200 larvae per pan), mean pupal survival as well as male and female mean adult dry weights were significantly reduced and median developmental times were significantly prolonged. However, excess per capita food did not allow low density cohorts (i.e., 100 larvae per pan) to increase adult production, developmental rate, or adult dry weight compared with cohorts reared under standard rearing conditions. Male and female pupal wet weights, adult dry weights, and adult wing lengths all were correlated for Ae. polynesiensis collected as pupae from natural habitats near Papara Commune, Tahiti. Mean adult dry weights of host-seeking females from the same and a neighboring location did not differ significantly from weights of females emerging from field-collected pupae. The comparison of mean adult dry weight of these adults with adults reared at different densities in the laboratory indicated that field populations develop under food-limited conditions. Aedes polynesiensis responds to intraspecific larval competition by producing small adults over elongated developmental periods. Pupal wet weights, adult dry weights, and adult wing lengths are equally acceptable measures of mosquito size for vector and fecundity studies.

International audience; The population structure of the Polynesian mosquito Aedes polynesiensis was investigated using electrophoretic data from two polymorphic protein loci. Considerable differentiation was observed both within and between islands in different archipelagos (Society, Tuamotu, Austral). Gene flow evaluated by F-st estimates was independent of geographic distance between islands but related to commercial traffic intensity. The results are discussed in view of recent findings on the variability of susceptibility to insecticides and of suitability as a vector for the nematode Wuchereria bancrofti.

Wolbachia is a maternally transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce pathogen interference and spread into mosquito vector populations makes it possible to develop Wolbachia as a biological control agent for vector-borne disease control. Although Wolbachia induces resistance to dengue virus (DENV), filarial worms, and Plasmodium in mosquitoes, species like Aedes polynesiensis and Aedes albopictus, which carry native Wolbachia infections, are able to transmit dengue and filariasis. In a previous study, the native wPolA in Ae. polynesiensis was replaced with wAlbB from Ae. albopictus, and resulted in the generation of the transinfected “MTB” strain with low susceptibility for filarial worms. In this study, we compare the dynamics of DENV serotype 2 (DENV-2) within the wild type “APM” strain and the MTB strain of Ae. polynesiensis by measuring viral infection in the mosquito whole body, midgut, head, and saliva at different time points post infection. The results show that wAlbB can induce a strong resistance to DENV-2 in the MTB mosquito. Evidence also supports that this resistance is related to a dramatic increase in Wolbachia density in the MTB's somatic tissues, including the midgut and salivary gland. Our results suggests that replacement of a native Wolbachia with a novel infection could serve as a strategy for developing a Wolbachia-based approach to target naturally infected insects for vector-borne disease control., Author Summary Aedes polynesiensis is a vector for both dengue and filariasis in the South Pacific. Efforts are ongoing to utilize Wolbachia as a biological control agent targeting this vector through either population suppression via releases of incompatible males or population replacement for spreading disease resistance into a population. Replacing the native Wolbachia with a novel infection from Ae. albopictus has generated the “MTB” strain of Ae. polynesiensis. This MTB mosquito is reproductively-incompatible with the wild type of Ae. polynesiensis and has a low susceptibility for filarial worms. In this work, we show that the MTB mosquito is resistant to dengue virus with a reduced viral infection in the mosquito whole body, midgut, head, and saliva. Our results further show its refractoriness to dengue virus is associated with a dramatic increase in Wolbachia density in those mosquito tissues where dengue virus needs to reside, replicate, and travel in order to be transmitted to humans. These results suggest that the MTB strain has the potential to be used in Wolbachia-based strategies to control both dengue and filariasis in the South Pacific.