Indoor resting behavior of Aedes aegypti (Diptera: Culicidae) in northeastern Thailand.

BACKGROUND: Aedes aegypti is a vector of several arboviruses, notably dengue virus (DENV), which causes dengue fever and is often found resting indoors. Culex spp. are largely nuisance mosquitoes but can include species that are vectors of zoonotic pathogens. Vector control is currently the main method to control dengue outbreaks. Indoor residual spraying can be part of an effective vector control strategy but requires an understanding of the resting behavior. Here we focus on the indoor-resting behavior of Ae. aegypti and Culex spp. in northeastern Thailand. METHODS: Mosquitoes were collected in 240 houses in rural and urban settings from May to August 2019 at two collection times (morning/afternoon), in four room types (bedroom, bathroom, living room and kitchen) in each house and at three wall heights (< 0.75 m, 0.75-1.5 m, > 1.5 m) using a battery-driven aspirator and sticky traps. Household characteristics were ascertained. Mosquitoes were identified as Ae. aegypti, Aedes albopictus and Culex spp. Dengue virus was detected in Ae. aegypti. Association analyses between urban/rural and within-house location (wall height, room), household variables, geckos and mosquito abundance were performed. RESULTS: A total of 2874 mosquitoes were collected using aspirators and 1830 using sticky traps. Aedes aegypti and Culex spp. accounted for 44.78% and 53.17% of the specimens, respectively. Only 2.05% were Ae. albopictus. Aedes aegypti and Culex spp. rested most abundantly at intermediate and low heights in bedrooms or bathrooms (96.6% and 85.2% for each taxon of the total, respectively). Clothes hanging at intermediate heights were associated with higher mean numbers of Ae. aegypti in rural settings (0.81 [SEM: 0.08] vs. low: 0.61 [0.08] and high: 0.32 [0.09]). Use of larval control was associated with lower numbers of Ae. aegypti (yes: 0.61 [0.08]; no: 0.70 [0.07]). All DENV-positive Ae. aegypti (1.7%, 5 of 422) were collected in the rural areas and included specimens with single, double and even triple serotype infections. CONCLUSIONS: Knowledge of the indoor resting behavior of adult mosquitoes and associated environmental factors can guide the choice of the most appropriate and effective vector control method. Our work suggests that vector control using targeted indoor residual spraying and/or potentially spatial repellents focusing on walls at heights lower than 1.5 m in bedrooms and bathrooms could be part of an integrated effective strategy for dengue vector control.

Considering waning immunity to better explain dengue dynamics.

Life-long serotype-specific immunity following dengue virus infection may not always occur, but the true extent of this effect is unknown. Analysis of more than 20 years of monotypic epidemics in the isolated French Polynesian islands revealed that whilst the risk of symptomatic dengue infection did conform to the classical paradigms of homotypic immunity and increased disease risk in heterotypic secondary infections, incorporation of waning immunity improved the ability of epidemiological models to capture the observed epidemic dynamics. Not only does this show how inclusion of waning immunity into classical models can reveal important facets of the immune response to natural dengue virus infection, it also has significant ramifications for vaccine development and implementation in dengue endemic areas.

Importance of Public Transport Networks for Reconciling the Spatial Distribution of Dengue and the Association of Socio-Economic Factors with Dengue Risk in Bangkok, Thailand.

Dengue is the most widespread mosquito-borne viral disease of man and spreading at an alarming rate. Socio-economic inequality has long been thought to contribute to providing an environment for viral propagation. However, identifying socio-economic (SE) risk factors is confounded by intra-urban daily human mobility, with virus being ferried across cities. This study aimed to identify SE variables associated with dengue at a subdistrict level in Bangkok, analyse how they explain observed dengue hotspots and assess the impact of mobility networks on such associations. Using meteorological, dengue case, national statistics, and transport databases from the Bangkok authorities, we applied statistical association and spatial analyses to identify SE variables associated with dengue and spatial hotspots and the extent to which incorporating transport data impacts the observed associations. We identified three SE risk factors at the subdistrict level: lack of education, % of houses being cement/brick, and number of houses as being associated with increased risk of dengue. Spatial hotspots of dengue were found to occur consistently in the centre of the city, but which did not entirely have the socio-economic risk factor characteristics. Incorporation of the intra-urban transport network, however, much improved the overall statistical association of the socio-economic variables with dengue incidence and reconciled the incongruous difference between the spatial hotspots and the SE risk factors. Our study suggests that incorporating transport networks enables a more real-world analysis within urban areas and should enable improvements in the identification of risk factors.

Viral transmissibility of SARS-CoV-2 accelerates in the winter, similarly to influenza epidemics.

The transmissibility of SARS-CoV-2 is anticipated to increase in the winter because of increased viral survival in cold damp air and thus would exacerbate viral spread in community. Analysis to capture the seasonal trend is needed to be prepared for future epidemics. We compared regression models for the 5-week case prior to each epidemic peak week for both the COVID-19 and influenza epidemics in winter and summer. The weekly case increase ratio was compared, using non-paired t tests between seasons. In order to test the robustness of seasonal transmission patterns, the normalized weekly case numbers of COVID-19 and influenza case rates of all seasons were assessed in a combined quadratic regression analysis. In winter, the weekly case increase ratio accelerated before epidemic peaks, similarly, for both COVID-19 and influenza. The quadratic regression models of weekly cases were observed to be convex curves in the winter and concave curves in the spring/summer for both COVID-19 and influenza. A significant increase of case increase ratio (3.19 [95%CI:0.01-6.37, P = .049]) of the COVID-19 and influenza epidemics was observed in winter as compared to spring/summer before the epidemic peak. The epidemic of COVID-19 was found to mirror that of influenza, suggesting a strong underlying seasonal transmissibility. Influenza epidemics can potentially be a useful reference for the COVID-19 epidemics.

Mapping the spatial distribution of the dengue vector Aedes aegypti and predicting its abundance in northeastern Thailand using machine-learning approach.

BACKGROUND: Mapping the spatial distribution of the dengue vector Aedes (Ae.) aegypti and accurately predicting its abundance are crucial for designing effective vector control strategies and early warning tools for dengue epidemic prevention. Socio-ecological and landscape factors influence Ae. aegypti abundance. Therefore, we aimed to map the spatial distribution of female adult Ae. aegypti and predict its abundance in northeastern Thailand based on socioeconomic, climate change, and dengue knowledge, attitude and practices (KAP) and/or landscape factors using machine learning (ML)-based system. METHOD: A total of 1066 females adult Ae. aegypti were collected from four villages in northeastern Thailand during January-December 2019. Information on household socioeconomics, KAP regarding climate change and dengue, and satellite-based landscape data were also acquired. Geographic information systems (GIS) were used to map the household-based spatial distribution of female adult Ae. aegypti abundance (high/low). Five popular supervised learning models, logistic regression (LR), support vector machine (SVM), k-nearest neighbor (kNN), artificial neural network (ANN), and random forest (RF), were used to predict females adult Ae. aegypti abundance (high/low). The predictive accuracy of each modeling technique was calculated and evaluated. Important variables for predicting female adult Ae. aegypti abundance were also identified using the best-fitted model. RESULTS: Urban areas had higher abundance of female adult Ae. aegypti compared to rural areas. Overall, study respondents in both urban and rural areas had inadequate KAP regarding climate change and dengue. The average landscape factors per household in urban areas were rice crop (47.4%), natural tree cover (17.8%), built-up area (13.2%), permanent wetlands (21.2%), and rubber plantation (0%), and the corresponding figures for rural areas were 12.1, 2.0, 38.7, 40.1 and 0.1% respectively. Among all assessed models, RF showed the best prediction performance (socioeconomics: area under curve, AUC = 0.93, classification accuracy, CA = 0.86, F1 score = 0.85; KAP: AUC = 0.95, CA = 0.92, F1 = 0.90; landscape: AUC = 0.96, CA = 0.89, F1 = 0.87) for female adult Ae. aegypti abundance. The combined influences of all factors further improved the predictive accuracy in RF model (socioeconomics + KAP + landscape: AUC = 0.99, CA = 0.96 and F1 = 0.95). Dengue prevention practices were shown to be the most important predictor in the RF model for female adult Ae. aegypti abundance in northeastern Thailand. CONCLUSION: The RF model is more suitable for the prediction of Ae. aegypti abundance in northeastern Thailand. Our study exemplifies that the application of GIS and machine learning systems has significant potential for understanding the spatial distribution of dengue vectors and predicting its abundance. The study findings might help optimize vector control strategies, future mosquito suppression, prediction and control strategies of epidemic arboviral diseases (dengue, chikungunya, and Zika). Such strategies can be incorporated into One Health approaches applying transdisciplinary approaches considering human-vector and agro-environmental interrelationships.

Development and Comparison of Dengue Vulnerability Indices Using GIS-Based Multi-Criteria Decision Analysis in Lao PDR and Thailand.

Dengue is a continuous health burden in Laos and Thailand. We assessed and mapped dengue vulnerability in selected provinces of Laos and Thailand using multi-criteria decision approaches. An ecohealth framework was used to develop dengue vulnerability indices (DVIs) that explain links between population, social and physical environments, and health to identify exposure, susceptibility, and adaptive capacity indicators. Three DVIs were constructed using two objective approaches, Shannon's Entropy (SE) and the Water-Associated Disease Index (WADI), and one subjective approach, the Best-Worst Method (BWM). Each DVI was validated by correlating the index score with dengue incidence for each spatial unit (district and subdistrict) over time. A Pearson's correlation coefficient (r) larger than 0.5 and a p-value less than 0.05 implied a good spatial and temporal performance. Spatially, DVIWADI was significantly correlated on average in 19% (4-40%) of districts in Laos (mean r = 0.5) and 27% (15-53%) of subdistricts in Thailand (mean r = 0.85). The DVISE was validated in 22% (12-40%) of districts in Laos and in 13% (3-38%) of subdistricts in Thailand. The DVIBWM was only developed for Laos because of lack of data in Thailand and was significantly associated with dengue incidence on average in 14% (0-28%) of Lao districts. The DVIWADI indicated high vulnerability in urban centers and in areas with plantations and forests. In 2019, high DVIWADI values were observed in sparsely populated areas due to elevated exposure, possibly from changes in climate and land cover, including urbanization, plantations, and dam construction. Of the three indices, DVIWADI was the most suitable vulnerability index for the study area. The DVIWADI can also be applied to other water-associated diseases, such as Zika and chikungunya, to highlight priority areas for further investigation and as a tool for prevention and interventions.

Social and environmental risk factors for dengue in Delhi city: A retrospective study.

Global urbanization is leading to an inexorable spread of several major diseases that need to be stemmed. Dengue is one of these major diseases spreading in cities today, with its principal mosquito vector superbly adapted to the urban environment. Current mosquito control strategies are proving inadequate, especially in the face of such urbanisation and novel, evidence-based targeted approaches are needed. Through combined epidemiological and entomological approaches, we aimed to identify a novel sanitation strategy to alleviate the burden of dengue through how the dengue virus spreads through the community. We combined surveillance case mapping, prospective serological studies, year-round mosquito surveys, socio-economic and Knowledge Attitudes and Practices surveys across Delhi. We identified lack of access to tap water (≤98%) as an important risk factor for dengue virus IgG sero-positivity (adjusted Odds Ratio 4.69, 95% C.I. 2.06-10.67) and not poverty per se. Wealthier districts had a higher dengue burden despite lower mosquito densities than the Intermediary income communities (adjusted Odds Ratio 2.92, 95% C.I. 1.26-6.72). This probably reflects dengue being introduced by people travelling from poorer areas to work in wealthier houses. These poorer, high density areas, where temperatures are also warmer, also had dengue cases during the winter. Control strategies based on improved access to a reliable supply of tap water plus focal intervention in intra-urban heat islands prior to the dengue season could not only lead to a reduction in mosquito abundance but also eliminate the reservoir of dengue virus clearly circulating at low levels in winter in socio-economically disadvantaged areas.

Knowledge, attitudes, and practices on climate change and dengue in Lao People's Democratic Republic and Thailand.

BACKGROUND: Dengue is linked with climate change in tropical and sub-tropical countries including the Lao People's Democratic Republic (Laos) and Thailand. Knowledge about these issues and preventive measures can affect the incidence and outbreak risk of dengue. Therefore, the present study was conducted to determine the knowledge, attitudes, and practices (KAP) among urban and rural communities and government officials about climate change and dengue in Laos and Thailand. METHODS: A cross-sectional KAP survey about climate change and dengue were conducted in 360 households in Laos (180 urban and 180 rural), 359 households in Thailand (179 urban and 180 rural), and 20 government officials (10 in each country) using structured questionnaires. Data analysis was undertaken using descriptive methods, principal component analysis (PCA), Chi-square test or Fisher's exact test (as appropriate), and logistic regression. RESULTS: Significant differences among the selected communities in both countries were found in terms of household participant's age, level of education, socioeconomic status, attitude level of climate change and KAP level of dengue (P < 0.05; 95% CI). Overall, participants' KAP about climate change and dengue were low except the attitude level for dengue in both countries. The level of awareness among government officials regarding the climatic relationship with dengue was also low. In Lao households, participants' knowledge about climate change and dengue was significantly associated with the level of education and socioeconomic status (SES) (P < 0.01). Their attitudes towards climate change and dengue were associated with educational level and internet use (P < 0.05). Householders' climate change related practices were associated with SES (P < 0.01) and dengue related practices were associated with educational level, SES, previous dengue experience and internet use (P < 0.01). In Thailand, participants' knowledge about climate change was associated with the level of education and SES (P < 0.01). Their attitudes towards climate change were associated with residence status (urban/rural) and internet use (P < 0.05); climate change related practices were associated with educational level and SES (P < 0.05). Dengue related knowledge of participants was associated with SES and previous dengue experience (P < 0.05); participants' dengue related attitudes and practices were associated with educational level (P < 0.01). CONCLUSION: The findings call for urgently needed integrated awareness programs to increase KAP levels regarding climate change adaptation, mitigation and dengue prevention to improve the health and welfare of people in these two countries, and similar dengue-endemic countries.

Efficacy of the In2Care® auto-dissemination device for reducing dengue transmission: study protocol for a parallel, two-armed cluster randomised trial in the Philippines.

BACKGROUND: Mosquito-borne viruses are imposing an ever increasing health burden worldwide. In addition to the recent Zika and chikungunya virus epidemics, dengue viruses have become the fastest growing problem with a 40-fold increase in the number of reported cases over the past five decades. Current mosquito control techniques involving larval source reduction, larviciding, and space spray of adulticides are costly, laborious, and of debatable efficacy. There remains an urgent need for the development of intervention methods that can be reasonably implemented in the context of modern day urbanisation. Auto-dissemination (AD) of insecticide by adult mosquitoes offers a potentially practical and useful tool in an integrated vector control programme. Recently, an immediately employable AD device, the In2Care® mosquito trap, has been commercialised and shows promise as an effective tool. However, there remains a lack of demonstration of epidemiological efficacy. METHODS/DESIGN: This trial aims to assess the extent to which implementation of In2Care® mosquito traps can reduce vector Aedes (Stegomyia) spp. adult mosquito densities and dengue virus transmission as measured by sequential sero-conversion rates in children 6-16 years of age in a dengue endemic location: Lipa City, Philippines. To achieve this, we will carry out a parallel, two-armed cluster randomised trial evaluating AD efficacy for reducing the incidence of dengue over a 2-year period with 4 consecutive months of vector control during peak dengue transmission each year. DISCUSSION: For decades, it has been commonly accepted that an integrated approach to mosquito control is required. The World Health Organization (WHO) Global Strategic Framework for Integrated Vector Management recommends a range of interventions, in combination, to increase control impact to reduce transmission. This efficacy trial of the first commercial product using the AD approach will be informative in assessing the general utility of AD in reducing not only adult vector densities but, more importantly, reducing the incidence of dengue. The AD technique may complement source reduction and larviciding campaigns by more efficiently targeting the most productive containers and those beyond human reach. If successful, this mosquito control strategy could prove an invaluable tool in the fight against urban mosquito vectors and a reduction in the burden of associated disease. TRIAL REGISTRATION: ISRCTN44272773 . Registered on 31 January 2019.

An evaluation of efficacy of the auto-dissemination technique as a tool for Aedes aegypti control in Madeira, Portugal.

BACKGROUND: The frequency and intensity of arboviral epidemics is steadily increasing and posing an intractable public health burden. Current vector control methods are proving ineffectual and despite progress in the development of high technology approaches, there is an urgent need for the development of tools for immediate implementation. Several studies suggest that the auto-dissemination of pyriproxyfen (PPF) is a promising new approach to larviciding although there is little detail on the conditions under which it is optimally effective. Here, we evaluate the efficacy of the approach in urban and rural sites in Madeira, Portugal. RESULTS: Auto-dissemination of PPF through adapted Biogents Sentinel traps (BGSTs) resulted in a modest but consistent impact on both juvenile and adult mosquito populations, but with considerable spatial heterogeneity. This heterogeneity was related to the distance from the BGST dissemination station as well as the local density of adult mosquitoes. There was evidence that the impact of PPF was cumulative over time both locally and with gradual spatial expansion. CONCLUSIONS: The density of adult mosquitoes and the spatial distribution of dissemination devices are key factors in mediating efficacy. In addition, urban topography may affect the efficiency of auto-dissemination by impeding adult mosquito dispersal. Further studies in a range of urban landscapes are necessary to guide optimal strategies for the implementation of this potentially efficacious and cost-effective approach to larviciding.

Global Vector Control Guidelines - The Need For Co-Creation.

In response to the global expansion and re-emergence of vector-borne diseases, WHO launched the Global Vector Control Response program in 2017. The aim is to strengthen vector control through increased capacity, innovation, improved surveillance, better coordination, and integrated action. Will this new approach overcome the failings of previous initiatives?

Estimating Frequency of Probable Autochthonous Cases of Dengue, Japan.

Imported dengue into naive areas is a recognized but unquantified threat. Differentiating imported and autochthonous cases remains problematic. A threshold approach applied to Japan identified several aberrant incidences of dengue. Despite these alerts, no epidemics occurred other than 1 in Yoyogi Park in Tokyo, which was probably an unusual event.

First identification of Rickettsia helvetica in questing ticks from a French Northern Brittany Forest.

Tick-borne rickettsiae are considered to be emerging, but data about their presence in western Europe are scarce. Ixodes ricinus ticks, the most abundant and widespread tick species in western Europe, were collected and tested for the presence of several tick-borne pathogens in western France, a region never previously explored in this context. There was a high tick abundance with a mean of 4 females, 4.5 males, and 23.3 nymphs collected per hour per collector. Out of 622 tested ticks, specific PCR amplification showed the presence of tick symbionts as well as low prevalence of Borrelia burgdorferi (0.8%), Bartonella spp. (0.17%), and Anaplasma phagocytophilum (0.09%). The most prevalent pathogen was Rickettsia helvetica (4.17%). This is the first time that this bacteria has been detected in ticks in this region, and this result raises the possibility that bacteria other than those classically implicated may be involved in rickettsial diseases in western France.

Environmental factors influencing tick densities over seven years in a French suburban forest.

BACKGROUND: Worldwide changes in socio-economic and environmental factors and the global climate are recognised causes of variation in tick distribution and density. Thus it is of great importance that new studies address the changing risk of infection for exposed populations. In Europe, Ixodes ricinus ticks are the most common vectors of several pathogens impacting veterinary and public health that have colonised suburban habitats. METHODS: This study aimed to evaluate longitudinal I. ricinus questing densities and infection rates over 7 years in a French suburban forested area with high human population density. Ticks were collected in spring yearly between 2008 and 2014 and, out of a total of 8594 collected I. ricinus, a representative subset of adult females (n = 259) were individually examined for the presence of several pathogens via PCR. RESULTS: Nymph densities peaked in 2009-2011, and then declined in 2012-2014. Changes in monthly temperature only had a modest impact on this variation. In contrast, analysis revealed a complex intra-annual relationship between mean nymph density and both concurrent and lagged mean monthly temperatures. The following pathogens were detected in the studied area: Anaplasma phagocytophilum, Rickettsia helvetica, Babesia venatorum and B. divergens, Francisella tularensis, Borrelia miyamotoi, B. afzelii/valaisiana, B. garinii/lusitaniae and Bartonella spp. CONCLUSION: Our findings reinforce the conclusion that ticks are important vectors of pathogenic microorganisms in suburban forests and suggest that despite complex intra-annual relationships between tick densities and temperature, there is no evidence for a climate-associated increase in infection risk over the 7-year period. Rather, tick densities are likely to be strongly influenced by population density fluctuations in vertebrate host species and wildlife management. Further detailed studies on the impact of climate change on tick population densities are required.

Determinants of Arbovirus Vertical Transmission in Mosquitoes.

Vertical transmission (VT) and horizontal transmission (HT) of pathogens refer to parental and non-parental chains of host-to-host transmission. Combining HT with VT enlarges considerably the range of ecological conditions in which a pathogen can persist, but the factors governing the relative frequency of each transmission mode are poorly understood for pathogens with mixed-mode transmission. Elucidating these factors is particularly important for understanding the epidemiology of arthropod-borne viruses (arboviruses) of public health significance. Arboviruses are primarily maintained by HT between arthropod vectors and vertebrate hosts in nature, but are occasionally transmitted vertically in the vector population from an infected female to her offspring, which is a proposed maintenance mechanism during adverse conditions for HT. Here, we review over a century of published primary literature on natural and experimental VT, which we previously assembled into large databases, to identify biological factors associated with the efficiency of arbovirus VT in mosquito vectors. Using a robust statistical framework, we highlight a suite of environmental, taxonomic, and physiological predictors of arbovirus VT. These novel insights contribute to refine our understanding of strategies employed by arboviruses to persist in the environment and cause substantial public health concern. They also provide hypotheses on the biological processes underlying the relative VT frequency for pathogens with mixed-mode transmission that can be tested empirically.

Urban climate versus global climate change-what makes the difference for dengue?

The expansion in the geographical distribution of vector-borne diseases is a much emphasized consequence of climate change, as are the consequences of urbanization for diseases that are already endemic, which may be even more important for public health. In this paper, we focus on dengue, the most widespread urban vector-borne disease. Largely urban with a tropical/subtropical distribution and vectored by a domesticated mosquito, Aedes aegypti, dengue poses a serious public health threat. Temperature plays a determinant role in dengue epidemic potential, affecting crucial parts of the mosquito and viral life cycles. The urban predilection of the mosquito species will further exacerbate the impact of global temperature change because of the urban heat island effect. Even within a city, temperatures can vary by 10 °C according to urban land use, and diurnal temperature range (DTR) can be even greater. DTR has been shown to contribute significantly to dengue epidemic potential. Unraveling the importance of within-city temperature is as important for dengue as for the negative health consequences of high temperatures that have thus far been emphasized, for example, pollution and heat stroke. Urban and landscape planning designed to mitigate the non-infectious negative effects of temperature should additionally focus on dengue, which is currently spreading worldwide with no signs of respite.

Plasmodium copy number variation scan: gene copy numbers evaluation in haploid genomes.

BACKGROUND: In eukaryotic genomes, deletion or amplification rates have been estimated to be a thousand more frequent than single nucleotide variation. In Plasmodium falciparum, relatively few transcription factors have been identified, and the regulation of transcription is seemingly largely influenced by gene amplification events. Thus copy number variation (CNV) is a major mechanism enabling parasite genomes to adapt to new environmental changes. METHODS: Currently, the detection of CNVs is based on quantitative PCR (qPCR), which is significantly limited by the relatively small number of genes that can be analysed at any one time. Technological advances that facilitate whole-genome sequencing, such as next generation sequencing (NGS) enable deeper analyses of the genomic variation to be performed. Because the characteristics of Plasmodium CNVs need special consideration in algorithms and strategies for which classical CNV detection programs are not suited a dedicated algorithm to detect CNVs across the entire exome of P. falciparum was developed. This algorithm is based on a custom read depth strategy through NGS data and called PlasmoCNVScan. RESULTS: The analysis of CNV identification on three genes known to have different levels of amplification and which are located either in the nuclear, apicoplast or mitochondrial genomes is presented. The results are correlated with the qPCR experiments, usually used for identification of locus specific amplification/deletion. CONCLUSIONS: This tool will facilitate the study of P. falciparum genomic adaptation in response to ecological changes: drug pressure, decreased transmission, reduction of the parasite population size (transition to pre-elimination endemic area).

The Spread of Dengue in an Endemic Urban Milieu--The Case of Delhi, India.

BACKGROUND: Dengue is a major international public health concern, one of the most important arthropod-borne diseases. More than 3.5 billion people are at risk of dengue infection and there are an estimated 390 million dengue infections annually. This prolific increase has been connected to societal changes such as population growth and increasing urbanization generating intense agglomeration leading to proliferation of synanthropic mosquito species. Quantifying the spatio-temporal epidemiology of dengue in large cities within the context of a Geographic Information System is a first step in the identification of socio-economic risk factors. METHODOLOGY/PRINCIPAL FINDINGS: This Project has been approved by the ethical committee of Institut Pasteur. Data has been anonymized and de-identified prior to geolocalisation and analysis. A GIS was developed for Delhi, enabling typological characterization of the urban environment. Dengue cases identified in the Delhi surveillance system from 2008 to 2010 were collated, localised and embedded within this GIS. The spatio-temporal distribution of dengue cases and extent of clustering were analyzed. Increasing distance from the forest in Delhi reduced the risk of occurrence of a dengue case. Proximity to a hospital did not increase risk of a notified dengue case. Overall, there was high heterogeneity in incidence rate within areas with the same socio-economical profiles and substantial inter-annual variability. Dengue affected the poorest areas with high density of humans, but rich areas were also found to be infected, potentially because of their central location with respect to the daily mobility network of Delhi. Dengue cases were highly clustered in space and there was a strong relationship between the time of introduction of the virus and subsequent cluster size. At a larger scale, earlier introduction predicted the total number of cases. CONCLUSIONS/SIGNIFICANCE: DENV epidemiology within Delhi has a forest fire signature. The stochastic nature of this invasion process likely smothers any detectable socio-economic risk factors. However, the significant finding that the size of the dengue case cluster depends on the timing of its emergence emphasizes the need for early case detection and implementation of effective mosquito control. A better understanding of the role of population mobility in contributing to dengue risk could also help focus control on areas at particular risk of dengue virus importation.