Arboviral Disease Outbreaks in the Pacific Islands Countries and Areas, 2014 to 2020: A Systematic Literature and Document Review.

Arthropod-borne diseases pose a significant public health threat, accounting for greater than 17% of infectious disease cases and 1 million deaths annually. Across Pacific Island countries and areas (PICs), outbreaks of dengue, chikungunya, and Zika are increasing in frequency and scale. Data about arbovirus outbreaks are incomplete, with reports sporadic, delayed, and often based solely on syndromic surveillance. We undertook a systematic review of published and grey literature and contacted relevant regional authorities to collect information about arboviral activity affecting PICs between October 2014 and June 2020. Our literature search identified 1176 unique peer-reviewed articles that were reduced to 25 relevant publications when screened. Our grey literature search identified 873 sources. Collectively, these data reported 104 unique outbreaks, including 72 dengue outbreaks affecting 19 (out of 22) PICs, 14 chikungunya outbreaks affecting 11 PICs, and 18 Zika outbreaks affecting 14 PICs. Our review is the most complete account of arboviral outbreaks to affect PICs since comparable work was published in 2014. It highlights the continued elevated level of arboviral activity across the Pacific and inconsistencies in how information about outbreaks is reported and recorded. It demonstrates the importance of a One-Health approach and the role that improved communication and reporting between different governments and sectors play in understanding the emergence, circulation, and transboundary risks posed by arboviral diseases.

Recent trends in global insecticide use for disease vector control and potential implications for resistance management.

Insecticides have played a major role in the prevention, control, and elimination of vector-borne diseases, but insecticide resistance threatens the efficacy of available vector control tools. A global survey was conducted to investigate vector control insecticide use from 2010 to 2019. Out of 140 countries selected as sample for the study, 87 countries responded. Also, data on ex-factory deliveries of insecticide-treated nets (ITNs) were analyzed. Insecticide operational use was highest for control of malaria, followed by dengue, leishmaniasis and Chagas disease. Vector control relied on few insecticide classes with pyrethroids the most used overall. Results indicated that IRS programs have been slow to react to detection of pyrethroid resistance, while proactive resistance management using insecticides with unrelated modes of action was generally weak. The intensive use of recently introduced insecticide products raised concern about product stewardship regarding the preservation of insecticide susceptibility in vector populations. Resistance management was weakest for control of dengue, leishmaniasis or Chagas disease. Therefore, it will be vital that vector control programs coordinate on insecticide procurement, planning, implementation, resistance monitoring, and capacity building. Moreover, increased consideration should be given to alternative vector control tools that prevent the development of insecticide resistance.

Challenges to implementation and strengthening of initial COVID-19 surveillance in Vanuatu: January-April 2020.

The Pacific island nation of Vanuatu is vulnerable to emerging infectious diseases, including epidemics and pandemics; chronic food and water insecurity; and natural hazards, including cyclones, earthquakes, tsunamis, landslides and flooding. In March 2020, the World Health Organization characterized the outbreak of novel coronavirus disease 2019 (COVID-19) as a global pandemic. By the end of April 2020, Vanuatu had reported no confirmed cases of COVID-19. Data from several sources are collected in Vanuatu's COVID-19 surveillance system to provide an overview of the situation, including data from case investigations and management, syndromic surveillance for influenza-like illness, hospital surveillance and laboratory surveillance. Review of data collected from January to the end of April 2020 suggests that there was no sustained increase in influenza-like illness in the community and no confirmed cases were identified. Lessons learnt from the early implementation of surveillance activities, the changing landscape of laboratory testing and pharmaceutical interventions, as well as the global experience, particularly in other Pacific island countries, will inform the refinement of COVID-19 surveillance activities in Vanuatu.

Lessons from COVID-19-free Vanuatu: intensive health operations for Phase 1 of repatriation and quarantine, May-July 2020.

International borders to Vanuatu closed on 23 March 2020 due to the global COVID-19 pandemic. In May-July 2020, the Government of Vanuatu focused on the safe and timely return of citizens and residents while ensuring Vanuatu remained COVID-19 free. Under Phase 1 of repatriation, between 27 May and 23 June 2020, 1522 people arrived in the capital, Port Vila, and were placed in compulsory government-mandated 14-day quarantine in 15 hotels. Pre-arrival health operations included collection of repatriate information, quarantine facility assessments, training for personnel supporting the process, and tabletop and functional exercises with live scenario simulations. During quarantine, health monitoring, mental health assessments and psychosocial support were provided. All repatriates completed 14 days of quarantine. One person developed symptoms consistent with COVID-19 during quarantine but tested negative. Overall health operations were considered a success despite logistical and resource challenges. Lessons learnt were documented during a health sector after-action review held on 22 July 2020. Key recommendations for improvement were to obtain timely receipt of repatriate information before travel, limit the number of repatriates received and avoid the mixing of "travel cohorts," ensure sufficient human resources are available to support operations while maintaining other essential services, establish a command and control structure for health operations, develop training packages and deliver them to all personnel supporting operations, and coordinate better with other sectors to ensure health aspects are considered. These recommendations were applied to further improve health operations for subsequent repatriation and quarantine, with Phase 2 commencing on 1 August 2020.

Case Definitions of Clinical Malaria in Children from Three Health Districts in the North Region of Cameroon.

Malaria endemicity in Cameroon greatly varies according to ecological environment. In such conditions, parasitaemia, which is associated with fever, may not always suffice to define an episode of clinical malaria. The evaluation of malaria control intervention strategies mostly consists of identifying cases of clinical malaria and is crucial to promote better diagnosis for accurate measurement of the impact of the intervention. We sought out to define and quantify clinical malaria cases in children from three health districts in the Northern region of Cameroon. A cohort study of 6,195 children aged between 6 and 120 months was carried out during the raining season (July to October) between 2013 and 2014. Differential diagnosis of clinical malaria was performed using the parasite density and axillary temperature. At recruitment, patients with malaria-related symptoms (fever [axillary temperature ≥ 37.5°C], chills, severe malaise, headache, or vomiting) and a malaria positive blood smear were classified under clinical malaria group. The malaria attributable fraction was calculated using logistic regression models. Plasmodium falciparum was responsible for over 91% of infections. Children from Pitoa health district had the highest number of asymptomatic infections (45.60%) compared to those from Garoua and Mayo Oulo. The most suitable cut-off for the association between parasite densities and fever was found among children less than 24 months. Overall, parasite densities that ranged above 3,200 parasites per µl of blood could be used to define the malaria attributable fever cases. In groups of children aged between 24 and 59 months and 60 and 94 months, the optimum cut-off parasite density was 6,400 parasites per µl of blood, while children aged between 95 and 120 months had a cut-off of 800 parasites per µl of blood. In the same ecoepidemiological zone, clinical malaria case definitions are influenced by age and location (health district) and this could be considered when evaluating malaria intervention strategies in endemic areas.

Implications of insecticide resistance for malaria vector control with long-lasting insecticidal nets: evidence from health facility data from Benin.

BACKGROUND: Insecticide-based interventions have averted more than 500 million malaria cases since 2000, but insecticide resistance in mosquitoes could bring about a rebound in disease and mortality. This study investigated whether insecticide resistance was associated with increased incidence of clinical malaria. METHODS: In an area of southern Benin with insecticide resistance and high use of insecticide-treated nets (ITNs), malaria morbidity and insecticide resistance were measured simultaneously in 30 clusters (villages or collections of villages) multiple times over the course of 2 years. Insecticide resistance frequencies were measured using the standard World Health Organization bioassay test. Malaria morbidity was measured by cases recorded at health facilities both in the whole population using routinely collected data and in a passively followed cohort of children under 5 years old. RESULTS: There was no evidence that incidence of malaria from routinely collected data was higher in clusters with resistance frequencies above the median, either in children aged under 5 (RR = 1.27 (95% CI 0.81-2.00) p = 0.276) or in individuals aged 5 or over (RR = 1.74 (95% CI 0.91-3.34) p = 0.093). There was also no evidence that incidence was higher in clusters with resistance frequencies above the median in the passively followed cohort (RR = 1.11 (0.52-2.35) p = 0.777). CONCLUSIONS: This study found no association between frequency of resistance and incidence of clinical malaria in an area where ITNs are the principal form of vector control. This may be because, as other studies have shown, ITNs continue to offer some protection from malaria even in the presence of insecticide resistance. Irrespective of resistance, nets provide only partial protection so the development of improved or supplementary vector control tools is required to reduce Africa's unacceptably high malaria burden.

The bionomics of the malaria vector Anopheles rufipes Gough, 1910 and its susceptibility to deltamethrin insecticide in North Cameroon.

BACKGROUND: Following the recent discovery of the role of Anopheles rufipes Gough, 1910 in human malaria transmission in the northern savannah of Cameroon, we report here additional information on its feeding and resting habits and its susceptibility to the pyrethroid insecticide deltamethrin. METHODS: From 2011 to 2015, mosquito samples were collected in 38 locations across Garoua, Mayo Oulo and Pitoa health districts in North Cameroon. Adult anophelines collected using outdoor clay pots, window exit traps and indoor spray catches were checked for feeding status, blood meal origin and Plasmodium circumsporozoite protein. The susceptibility of field-collected An. rufipes to deltamethrin was assessed using WHO standard procedures. RESULTS: Of 9327 adult Anopheles collected in the 38 study sites, An. rufipes (6.5%) was overall the fifth most abundant malaria vector species following An. arabiensis (52.4%), An. funestus (s.l.) (20.8%), An. coluzzii (12.6%) and An. gambiae (6.8%). This species was found outdoors (51.2%) or entering houses (48.8%) in 35 suburban and rural locations, together with main vector species. Apart from human blood with index of 37%, An. rufipes also fed on animals including cows (52%), sheep (49%), pigs (16%), chickens (2%) and horses (1%). The overall parasite infection rate of this species was 0.4% based on the detection of P. falciparum circumsporozoite proteins in two of 517 specimens tested. Among the 21 An. rufipes populations assessed for deltamethrin susceptibility, seven populations were classified as "susceptible" (mortality ≥ 98%) , ten as "probable resistant" with a mortality range of 90-97% and four as "resistant" with a mortality range of 80-89%. CONCLUSIONS: This study revealed changeable resting and feeding behaviour of An. rufipes, as well as further evidence on its ability to carry human malaria parasites in North Cameroon. Besides, this species is developing physiological resistance to deltamethrin insecticide which is used in treated nets and agriculture throughout the country, and should be regarded as one of potential targets for the control of residual malaria parasite transmission in Africa.

Temporal and spatial trends in insecticide resistance in Anopheles arabiensis in Sudan: outcomes from an evaluation of implications of insecticide resistance for malaria vector control.

BACKGROUND: Long-lasting insecticidal nets (LLINs) (with pyrethroids) and indoor residual spraying (IRS) are the cornerstones of the Sudanese malaria control program. Insecticide resistance to the principal insecticides in LLINs and IRS is a major concern. This study was designed to monitor insecticide resistance in Anopheles arabiensis from 140 clusters in four malaria-endemic areas of Sudan from 2011 to 2014. All clusters received LLINs, while half (n = 70), distributed across the four regions, had additional IRS campaigns. METHODS: Anopheles gambiae (s.l.) mosquitoes were identified to species level using PCR techniques. Standard WHO insecticide susceptibility bioassays were carried out to detect resistance to deltamethrin (0.05%), DDT (4%) and bendiocarb (0.1%). TaqMan assays were performed on random samples of deltamethrin-resistant phenotyped and pyrethrum spray collected individuals to determine Vgsc-1014 knockdown resistance mutations. RESULTS: Anopheles arabiensis accounted for 99.9% of any anopheline species collected across all sites. Bioassay screening indicated that mosquitoes remained susceptible to bendiocarb but were resistance to deltamethrin and DDT in all areas. There were significant increases in deltamethrin resistance over the four years, with overall mean percent mortality to deltamethrin declining from 81.0% (95% CI: 77.6-84.3%) in 2011 to 47.7% (95% CI: 43.5-51.8%) in 2014. The rate of increase in phenotypic deltamethrin-resistance was significantly slower in the LLIN + IRS arm than in the LLIN-only arm (Odds ratio 1.34; 95% CI: 1.02-1.77). The frequency of Vgsc-1014F mutation varied spatiotemporally with highest frequencies in Galabat (range 0.375-0.616) and New Halfa (range 0.241-0.447). Deltamethrin phenotypic-resistance correlated with Vgsc-1014F frequency. CONCLUSION: Combining LLIN and IRS, with different classes of insecticide, may delay pyrethroid resistance development, but the speed at which resistance develops may be area-specific. Continued monitoring is vital to ensure optimal management and control.

Role of Anopheles (Cellia) rufipes (Gough, 1910) and other local anophelines in human malaria transmission in the northern savannah of Cameroon: a cross-sectional survey.

BACKGROUND: As part of a study to determine the impact of insecticide resistance on the effectiveness of long-lasting insecticide treated nets (LLINs) in the north of Cameroon, the unexpectedly high density and anthropophilic behaviour of Anopheles rufipes lead us to investigate this species bionomics and role in human malaria parasite transmission. METHODS: For four consecutive years (2011-2014), annual cross-sectional sampling of adult mosquitoes was conducted during the peak malaria season (September-October) in three health districts in northern Cameroon. Mosquitoes sampled by human landing catch and pyrethrum spray catch methods were morphologically identified, their ovaries dissected for parity determination and Anopheles gambiae siblings were identified by molecular assay. Infection with P. falciparum and blood meal source in residual fauna of indoor resting anopheline mosquitoes were determined by enzyme-linked-immunosorbent assays. RESULTS: Anopheles gambiae (sensu lato) (s.l.) comprised 18.4% of mosquitoes collected with An. arabiensis representing 66.27% of the sibling species. The proportion of An. rufipes (2.7%) collected was high with a human-biting rate ranging between 0.441 and 11.083 bites/person/night (b/p/n) and an anthropophagic rate of 15.36%. Although overall the members of An. gambiae complex were responsible for most of the transmission with entomological inoculation rates (EIR) reaching 1.221 infective bites/person/night (ib/p/n), An. arabiensis and An. coluzzii were the most implicated. The roles of An. funestus, An. pharoensis and An. paludis were minor. Plasmodium falciparum circumsporozoite protein rate in Anopheles rufipes varied from 0.6 to 5.7% with EIR values between 0.010 and 0.481 ib/p/n. CONCLUSIONS: The study highlights the epidemiological role of An. rufipes alongside the members of the An. gambiae complex, and several other sympatric species in human malaria transmission during the wet season in northern Cameroon. For the first time in Cameroon, An. rufipes has been shown to be an important local malaria vector, emphasising the need to review the malaria entomological profile across the country as pre-requisite to effective vector management strategies.

Implementation of the global plan for insecticide resistance management in malaria vectors: progress, challenges and the way forward.

In recent years, there has been an increase in resistance of malaria vectors to insecticides, particularly to pyrethroids which are widely used in insecticide-treated nets. The Global Plan for Insecticide Resistance Management in malaria vectors (GPIRM), released in May 2012, is a collective strategy for the malaria community to tackle this challenge. This review outlines progress made to date and the challenges experienced in the implementation of GPIRM, and outlines focus areas requiring urgent attention. Whilst there has been some advancement, uptake of GPIRM at the national level has generally been poor for various reasons, including limited availability of vector control tools with new mechanisms of action as well as critical financial, human and infrastructural resource deficiencies. There is an urgent need for a global response plan to address these deficits and ensure the correct and efficient use of available tools in order to maintain the effectiveness of current vector control efforts whilst novel vector control tools are under development. Emphasis must be placed on enhancing national capacities (such as human and infrastructural resources) to enable efficient monitoring and management of insecticide resistance, and to support availability and accessibility of appropriate new vector control products. Lack of action by the global community to address the threat of insecticide resistance is unacceptable and deprives affected communities of their basic right of universal access to effective malaria prevention. Aligning efforts and assigning the needed resources will ensure the optimal implementation of GPIRM with the ultimate goal of maintaining effective malaria vector control.

Malaria vector control at a crossroads: public health entomology and the drive to elimination.

Vector control has been at the core of successful malaria control. However, a dearth of field-oriented vector biologists threatens to undermine global reductions in malaria burden. Skilled cadres are needed to manage insecticide resistance, to maintain coverage with current interventions, to develop new paradigms for tackling 'residual' transmission and to target interventions as transmission becomes increasingly heterogeneous. Recognising this human resource crisis, in September 2013, WHO Global Malaria Programme issued guidance for capacity building in entomology and vector control, including recommendations for countries and implementing partners. Ministries were urged to develop long-range strategic plans for building human resources for public health entomology and vector control (including skills in epidemiology, geographic information systems, operational research and programme management) and to set in place the requisite professional posts and career opportunities. Capacity building and national ownership in all partner projects and a clear exit strategy to sustain human and technical resources after project completion were emphasised. Implementing partners were urged to support global and regional efforts to enhance public health entomology capacity. While the challenges inherent in such capacity building are great, so too are the opportunities to establish the next generation of public health entomologists that will enable programmes to continue on the path to malaria elimination.

An online tool for mapping insecticide resistance in major Anopheles vectors of human malaria parasites and review of resistance status for the Afrotropical region.

BACKGROUND: Malaria control programmes across Africa and beyond are facing increasing insecticide resistance in the major anopheline vectors. In order to preserve or prolong the effectiveness of the main malaria vector interventions, up-to-date and easily accessible insecticide resistance data that are interpretable at operationally-relevant scales are critical. Herein we introduce and demonstrate the usefulness of an online mapping tool, IR Mapper. METHODS: A systematic search of published, peer-reviewed literature was performed and Anopheles insecticide susceptibility and resistance mechanisms data were extracted and added to a database after a two-level verification process. IR Mapper ( http://www.irmapper.com) was developed using the ArcGIS for JavaScript Application Programming Interface and ArcGIS Online platform for exploration and projection of these data. RESULTS: Literature searches yielded a total of 4,084 susceptibility data points for 1,505 populations, and 2,097 resistance mechanisms data points for 1,000 populations of Anopheles spp. tested via recommended WHO methods from 54 countries between 1954 and 2012. For the Afrotropical region, data were most abundant for populations of An. gambiae, and pyrethroids and DDT were more often used in susceptibility assays (51.1 and 26.8% of all reports, respectively) than carbamates and organophosphates. Between 2001 and 2012, there was a clear increase in prevalence and distribution of confirmed resistance of An. gambiae s.l. to pyrethroids (from 41 to 87% of the mosquito populations tested) and DDT (from 64 to 91%) throughout the Afrotropical region. Metabolic resistance mechanisms were detected in western and eastern African populations and the two kdr mutations (L1014S and L1014F) were widespread. For An. funestus s.l., relatively few populations were tested, although in 2010-2012 resistance was reported in 50% of 10 populations tested. Maps are provided to illustrate the use of IR Mapper and the distribution of insecticide resistance in malaria vectors in Africa. CONCLUSIONS: The increasing pyrethroid and DDT resistance in Anopheles in the Afrotropical region is alarming. Urgent attention should be afforded to testing An. funestus populations especially for metabolic resistance mechanisms. IR Mapper is a useful tool for investigating temporal and spatial trends in Anopheles resistance to support the pragmatic use of insecticidal interventions.

Synergist bioassays: A simple method for initial metabolic resistance investigation of field Anopheles gambiae s.l. populations.

Metabolic resistance and the potential role of permeability-glycoprotein (P-gp) efflux pumps were investigated in a pyrethroid-resistant wild Anopheles gambiae s.l. Tiassalé population, using WHO susceptibility assays with deltamethrin (0.05%), with and without pre-exposure to synergists. The synergists used included an inhibitor of P-glycoprotein efflux pumps (verapamil), an inhibitor of esterases (EN 16-5), and an inhibitor of P450s and esterases (piperonyl butoxide). Pre-exposure to verapamil followed by deltamethrin led to a slight but non-significant (P=0.59) increase in mortality relative to exposure to deltamethrin alone (64.5% versus 69.2%). Similarly, pre-exposure to EN 16-5 yielded a non-significant increase in mortality (to 76.6%; P=0.85) but a significant increase in the knock down rate (from 48.3% to 78.7%; P<0.01). Pre-exposure with PBO caused a significant increase in mortality (to 93.1%; P<0.001) and knockdown rate (100%; P<0.001), which related to a 2.9 fold decrease in the resistance level. The results provide evidence that metabolic resistance mechanisms are present within the assessed mosquito population. The decrease in time to knock down of this population with deltamethrin following exposure to EN16-5 and PBO is of particular relevance to vector control, where quick knock down is a highly desired characteristic. The suspected resistance mechanisms present in this population merit further investigation through biochemical and molecular analyses for full resistance profile characterization. Bioassays with synergists can provide a quick and easy basis for initial characterization of resistant mosquito populations, without the need of preserved specimens, expensive equipment and substrates or specialized expertise.

Increase in susceptibility to insecticides with aging of wild Anopheles gambiae mosquitoes from Côte d'Ivoire.

BACKGROUND: Appropriate monitoring of vector insecticide susceptibility is required to provide the rationale for optimal insecticide selection in vector control programs. METHODS: In order to assess the influence of mosquito age on susceptibility to various insecticides, field-collected larvae of An. gambiae s.l. from Tiassalé were reared to adults. Females aged 1, 2, 3, 5 and 10 days were exposed to 5 insecticides (deltamethrin, permethrin, DDT, malathion and propoxur) using WHO susceptibility test kits. Outcome measures included the LT50 (exposure time required to achieve 50% knockdown), the RR (resistance ratio, i.e. a calculation of how much more resistant the wild population is compared with a standard susceptible strain) and the mortality rate following 1 hour exposure, for each insecticide and each mosquito age group. RESULTS: There was a positive correlation between the rate of knockdown and mortality for all the age groups and for all insecticides tested. For deltamethrin, the RR50 was highest for 2 day old and lowest for 10 day old individuals. Overall, mortality was lowest for 2 and 3 day old individuals and significantly higher for 10 day old individuals (P < 0.05). With permethrin, the RR50 was highest for 1 to 3 day old individuals and lowest for 10 day old individuals and mortality was lowest for 1 to 3 day old individuals, intermediate for 5 day old and highest for 10 day old individuals. DDT did not display any knockdown effect and mortality was low for all mosquito age groups (<7%). With malathion, the RR50 was low (1.54 - 2.77) and mortality was high (>93%) for all age groups. With propoxur, no knockdown effect was observed for 1, 2 and 3 day old individuals and a very low level of mortality was observed (< 4%), which was significantly higher for 5 and 10 day old individuals (30%, P < 0.01). CONCLUSION: Results indicate that for An. gambiae s.l. adults derived from wild-collected larvae, there was an influence of age on insecticide susceptibility status, with younger individuals (1 to 3 days old) more resistant than older mosquitoes. This indicates that the use of 1 - 2 day old mosquitoes in susceptibility assays as recommended by the WHO should facilitate detection of resistance at the stage where the highest rate of the resistance phenotype is present.

Quantitative relationships between immature and emergent adult Aedes aegypti (Diptera: Culicidae) populations in water storage container habitats.

Although quantitative surveillance data for immature stages of Aedes aegypti are often used to prioritize containers or specific types of containers for control, the relationship between immature and emergent adult populations under field conditions is largely unknown. We examined the relationships between abundance of III/IV instars and pupae, and emerging adult population for a series of water storage containers in southern Vietnam. A large proportion of III/IV instars failed to progress to adulthood, and the relationships between III/IV instars and adults were poor. Collected IV instars appeared to be nutritionally deprived, although their size and nutrient levels were not reliable indicators of emergence success. Conversely, pupal abundance was a good indicator of emerging adult populations, especially over the ensuing 48-h period. Although there were clear advantages of pupal surveillance over surveillance of III/IV instars for the estimation of adult mosquito productivity, there were practical limitations associated with the enumeration of pupae, and their comparatively low densities may preclude the identification of potentially productive containers.

Shifting priorities in vector biology to improve control of vector-borne disease.

Vector control remains the primary measure available to prevent pathogen transmission for the most devastating vector-borne diseases (VBDs): malaria, dengue, trypanosomiasis, filariasis, leishmaniasis, and Chagas disease. Current control strategies, however, are proving insufficient and the remarkable advances in the molecular biology of disease vectors over the last two decades have yet to result in tangible tools that effectively reduce VBD incidence. Here we argue that vector biologists must fundamentally shift their approach to VBD research. We propose an agenda highlighting the most critical avenues to improve the effectiveness of vector control. Research priorities must be diversified to support simultaneous development of multiple, alternative control strategies. Knowledge across relevant diseases and disciplines should be better integrated and disease prevention efforts extended beyond the academic sector to involve private industry, ministries of health, and local communities. To obtain information of more immediate significance to public health, the research focus must shift from laboratory models to natural pathogen-transmission systems. Identification and characterization of heterogeneities inherent to VBD systems should be prioritized to allow development of local, adaptive control strategies that efficiently make use of limited resources. Importantly, increased involvement of disease-endemic country (DEC) scientists, institutes, and communities will be key to enhance and sustain the fight against VBD.

Critical evaluation of quantitative sampling methods for Aedes aegypti (Diptera: Culicidae) immatures in water storage containers in Vietnam.

In response to an identified paucity of information on the size and composition of immature Aedes aegypti (L.) (Diptera: Culicidae) populations in large field containers, we assessed net sampling and pumping/sieving methods for estimating and enumerating third (III)/fourth (IV) instar and pupal populations. Sweep net detection thresholds (number above which > or = 90% chance of a positive sample) were < or = 28 immatures for seven different container types (115-3000 liter jars and tanks) in the laboratory, and mean recovery percentages varied by container type (6.15-41.29 and 7.39-33.10% for III/IV instars and pupae, respectively). A pumping method or hand bailing was applied in the field for the collection of III/IV instars and pupae from 406 receptacles, of which 343 had been previously sampled via a five-sweep netting technique. Larvae were 9.30 times more prevalent than pupae, and abundance varied by container type with means of 36-537 III/IV instars and 6-53 pupae per receptacle. Sweep netting for III/IV instars effectively identified 86.2% of Ae. aegypti-positive containers, whereas sampling for pupae detected only 43.1% of positive containers. When conversion factors (inverse of laboratory recovery percentages) were applied to field net sampling data, estimates of container populations were more accurate for III/IV instars than pupae (maximum R2 = 0.610 and 0.328, respectively); however, the relationship between immature abundance and emergent adult populations remains to be defined.

Entomological investigations in a focus of dengue transmission in Cairns, Queensland, Australia, by using the sticky ovitraps.

Sticky ovitraps (patent pending) were used to sample female Aedes aegypti (L.) weekly in a focus of dengue activity in Cairns, Queensland, Australia. In February 2003, transmission of dengue virus serotype 2 began in the suburb of Parramatta Park, peaking in mid-March 2003. This suburb features many older, unscreened houses with high populations of Ae. aegypti. Highest densities (2-3.5 females per trap per week) were obtained during peak dengue transmission (January and February) before mosquito control was initiated. Beginning in late March, female Ae. aegypti collected in sticky ovitraps were tested for dengue viral RNA by using a TaqMan reverse transcription-polymerase chain reaction assay. Dengue viral RNA was detected in six pools of Ae. aegypti collected in late March. The highest minimum infection rate was 116/1000 mosquitoes. After the initiation of larval control (containers treated with S-methoprene or lambda-cyhalothrin) and adult control (interior harborage sites sprayed with lambda-cyhalothrin) in early March, trap collections dropped to <0.5 per trap per week, and no virus was detected in trapped mosquitoes. Human cases subsequently dropped from a high of seven cases per day in mid-March to only sporadic cases in late April, with the final reported onset of 7 May. Sticky ovitraps have potential as a monitoring device for gravid Ae. aegypti and can be used to assess control efficacy and dengue virus activity. A sticky ovitrap index (mean number of female Ae. Aegypti per trap per week) could be useful in gauging the risk of dengue transmission.