Effects of seasonality and developed land cover on Culex mosquito abundance and microbiome diversity.

The vectorial capacity of mosquitoes, which influences the dynamics of vector-borne disease transmission, is intricately linked to mosquito abundance and the composition and diversity of their associated microbiomes. However, the influence of environmental factors on mosquito populations and microbiome diversity remains underexplored. Here we examined the effects of seasonality and developed land cover on Culex mosquito abundance and bacterial diversity. Biweekly field sampling of female Culex mosquitoes was conducted using gravid and CDC light traps, spanning summer to autumn across varying developed land cover levels in two urban areas in Central Illinois. Mosquito abundance was assessed by the number of mosquitoes captured per trap night and compared across seasons and developed levels. The mean mosquito abundance for gravid and light traps was 12.96 ± 2.15 and 7.67 ± 1.44, respectively. Notably, higher levels of developed land cover exhibited higher Culex abundance than the low level for light traps, but no significant difference was found between summer and early autumn. In gravid traps, no significant differences were detected across seasons or developed levels. Microbial analysis of the mosquito microbiome revealed that Proteobacteria and Wolbachia, with a mean relative abundance of 80.77 and 52.66% respectively, were identified as the most dominant bacterial phylum and genus. Their relative abundance remained consistent across seasons and developed land cover levels, with negligible variations. Alpha diversity, as measured by observed species, Chao1, Shannon, and Simpson, showed slightly higher values in early-autumn compared to late-summer. A notable pattern of bacterial diversity, as indicated by all four diversity indexes, is evident across varying levels of land development. Significantly, high or intermediate developed levels consistently showed reduced alpha diversity when compared to the lower level. This underscores the pronounced impact of anthropogenic ecological disturbances in shaping mosquito microbiomes. Beta diversity analysis revealed no significant dissimilarities in bacterial community composition across seasons and developed levels, although some separation was noted among different levels of developed land cover. These findings highlight the significant role of environmental factors in shaping mosquito abundance and their associated microbiomes, with potential implications for the vectorial capacity in the transmission of vector-borne diseases.

Larval diet and temperature alter mosquito immunity and development: using body size and developmental traits to track carry-over effects on longevity.

BACKGROUND: Estimating arbovirus transmission potential requires a mechanistic understanding of how environmental factors influence the expression of adult mosquito traits. While preimaginal exposure to environmental factors can have profound effects on adult traits, tracking and predicting these effects remains challenging. METHODS: Using Aedes albopictus and a structural equation modeling approach, we explored how larval nutrition and temperature jointly affect development rate and success, female body size, and whether these metrics capture carry-over effects on adult female longevity. Additionally, we investigated how larval diet and temperature affect the baseline expression of 10 immune genes. RESULTS: We found that larval development success was primarily determined by diet, while temperature and diet both affected development rate and female body size. Under a low larval diet, pupal wet weight and wing length both declined with increasing temperature. In contrast, responses of the two morphometric measures to rearing temperature diverged when females were provided higher larval nutrition, with pupal wet weight increasing and wing length decreasing at higher temperatures. Our analyses also revealed opposing relationships between adult female lifespan and the two morphometric measures, with wing length having a positive association with longevity and pupal weight a negative association. Larval diet indirectly affected adult longevity, and the time to pupation was negatively correlated with longevity. The expression of eight immune genes from the toll, JAK-STAT and Imd pathways was enhanced in mosquitoes with higher nutrition. CONCLUSIONS: Our results highlight deficiencies from using a single body size measure to capture carry-over effects on adult traits. Further studies of larval development rate under varying environmental conditions and its potential for tracking carry-over effects on vectorial capacity are warranted.

Nutritional stress compromises mosquito fitness and antiviral immunity, while enhancing dengue virus infection susceptibility.

Diet-induced nutritional stress can influence pathogen transmission potential in mosquitoes by impacting life history traits, infection susceptibility, and immunity. To investigate these effects, we manipulate mosquito diets at larval and adult stages, creating two nutritional levels (low and normal), and expose adults to dengue virus (DENV). We observe that egg number is reduced by nutritional stress at both stages and viral exposure separately and jointly, while the likelihood of laying eggs is exclusively influenced by adult nutritional stress. Adult nutritional stress alone shortens survival, while any pairwise combination between both-stage stress and viral exposure have a synergistic effect. Additionally, adult nutritional stress increases susceptibility to DENV infection, while larval nutritional stress likely has a similar effect operating via smaller body size. Furthermore, adult nutritional stress negatively impacts viral titers in infected mosquitoes; however, some survive and show increased titers over time. The immune response to DENV infection is overall suppressed by larval and adult nutritional stress, with specific genes related to Toll, JAK-STAT, and Imd immune signaling pathways, and antimicrobial peptides being downregulated. Our findings underscore the importance of nutritional stress in shaping mosquito traits, infection outcomes, and immune responses, all of which impact the vectorial capacity for DENV transmission.

Environmental Drivers of Gulf Coast Tick (Acari: Ixodidae) Range Expansion in the United States.

In the United States, the Gulf Coast tick (Amblyomma maculatum Koch) is a species of growing medical and veterinary significance, serving as the primary vector of the pathogenic bacterium, Rickettsia parkeri (Rickettsiales: Rickettsiaceae), in humans and the apicomplexan parasite, Hepatozoon americanum, in canines. Ongoing reports of A. maculatum from locations outside its historically reported distribution in the southeastern United States suggest the possibility of current and continuing range expansion. Using an ecological niche modeling approach, we combined new occurrence records with high-resolution climate and land cover data to investigate environmental drivers of the current distribution of A. maculatum in the United States. We found that environmental suitability for A. maculatum varied regionally and was primarily driven by climatic factors such as annual temperature variation and seasonality of precipitation. We also found that presence of A. maculatum was associated with open habitat with minimal canopy cover. Our model predicts large areas beyond the current distribution of A. maculatum to be environmentally suitable, suggesting the possibility of future northward and westward range expansion. These predictions of environmental suitability may be used to identify areas at potential risk for establishment and to guide future surveillance of A. maculatum in the United States.

Highlights of Medical Entomology, 2020.

Medical Entomology as a field is inherently global - thriving on international and interdisciplinary collaborations and affected dramatically by arthropod and pathogen invasions and introductions. This past year also will be remembered as the year in which the SARS-CoV-2 COVID-19 pandemic affected every part of our lives and professional activities and impacted (or changed, sometimes in good ways) our ability to collaborate and detect or respond to invasions. This incredible year is the backdrop for the 2020 Highlights in Medical Entomology. This article highlights the broad scope of approaches and disciplines represented in the 2020 published literature, ranging from sensory and chemical ecology, population genetics, impacts of human-mediated environmental change on vector ecology, life history and the evolution of vector behaviors, to the latest developments in vector surveillance and control.

Comparing Contributions of Passive and Active Tick Collection Methods to Determine Establishment of Ticks of Public Health Concern Within Illinois.

In Illinois, between 1990 and 2017, tick-borne diseases in humans increased 10-fold, yet we have insufficient information on when and where people are exposed to vector ticks (Ixodida: Ixodidae). The aims of our research were to compare contributions of passive and active tick collection methods in determining establishment of ticks of public health concern and obtain information on tick distributions within Illinois. We used three surveillance strategies within the Illinois Tick Inventory Collaboration Network to gather information about the ticks of public health concern: 1) passive collection (voluntary submission by the public); 2) systematic collection (biweekly active surveillance); and 3) special collections (active collections in locations of special interest). Of collected adult and nymphal ticks, 436 were from passive collections, 142 from systematic collections, and 1,270 from special collections. Tick species distribution status changed in 36 counties. Our data provide noteworthy updates to distribution maps for use by public health agencies to develop prevention and control strategies. Additionally, the program built a network of collaborations and partnerships to support future tick surveillance efforts within Illinois and highlighted how the combination of the three surveillance strategies can be used to determine geographic spread of ticks, pinpoint locations in need of more surveillance, and help with long-term efforts that support phenology studies.

Differential effects of larval and adult nutrition on female survival, fecundity, and size of the yellow fever mosquito, Aedes aegypti.

BACKGROUND: The yellow fever mosquito, Aedes aegypti, is the principal vector of medically-important infectious viruses that cause severe illness such as dengue fever, yellow fever and Zika. The transmission potential of mosquitoes for these arboviruses is largely shaped by their life history traits, such as size, survival and fecundity. These life history traits, to some degree, depend on environmental conditions, such as larval and adult nutrition (e.g., nectar availability). Both these types of nutrition are known to affect the energetic reserves and life history traits of adults, but whether and how nutrition obtained during larval and adult stages have an interactive influence on mosquito life history traits remains largely unknown. RESULTS: Here, we experimentally manipulated mosquito diets to create two nutritional levels at larval and adult stages, that is, a high or low amount of larval food (HL or LL) during larval stage, and a good and poor adult food (GA or PA, represents normal or weak concentration of sucrose) during adult stage. We then compared the size, survival and fecundity of female mosquitoes reared from these nutritional regimes. We found that larval and adult nutrition affected size and survival, respectively, without interactions, while both larval and adult nutrition influenced fecundity. There was a positive relationship between fecundity and size. In addition, this positive relationship was not affected by nutrition. CONCLUSIONS: These findings highlight how larval and adult nutrition differentially influence female mosquito life history traits, suggesting that studies evaluating nutritional effects on vectorial capacity traits should account for environmental variation across life stages.

Culex pipiens and Culex restuans egg rafts harbor diverse bacterial communities compared to their midgut tissues.

BACKGROUND: The bacterial communities associated with mosquito eggs are an essential component of the mosquito microbiota, yet there are few studies characterizing and comparing the microbiota of mosquito eggs to other host tissues. METHODS: We sampled gravid female Culex pipiens L. and Culex restuans Theobald from the field, allowed them to oviposit in the laboratory, and characterized the bacterial communities associated with their egg rafts and midguts for comparison through MiSeq sequencing of the 16S rRNA gene. RESULTS: Bacterial richness was higher in egg rafts than in midguts for both species, and higher in Cx pipiens than Cx. restuans. The midgut samples of Cx. pipiens and Cx. restuans were dominated by Providencia. Culex pipiens and Cx. restuans egg rafts samples were dominated by Ralstonia and Novosphingobium, respectively. NMDS ordination based on Bray-Curtis distance matrix revealed that egg-raft samples, or midgut tissues harbored similar bacterial communities regardless of the mosquito species. Within each mosquito species, there was a distinct clustering of bacterial communities between egg raft and midgut tissues. CONCLUSION: These findings expand the list of described bacterial communities associated with Cx. pipiens and Cx. restuans and the additional characterization of the egg raft bacterial communities facilitates comparative analysis of mosquito host tissues, providing a basis for future studies seeking to understand any functional role of the bacterial communities in mosquito biology.

Spatial, Temporal, and Genetic Invasion Dynamics of Aedes albopictus (Diptera: Culicidae) in Illinois.

The spread of the Asian tiger mosquito, Aedes albopictus Skuse, throughout the United States has implications for the transmission potential of vector-borne diseases. We used a 30-yr data set of occurrence records in Illinois and developed a hierarchical Bayesian model to shed light on the patterns and processes involved in the introduction and expansion along the northern edge of the geographic range of this species. We also collected specimens from 10 locations and sequenced a segment of their mitochondrial COI genes to assess possible introduction sources and geographic patterns in genetic variation present within contemporary populations. We documented an increase in the number of observations throughout the southern and central parts of Illinois over the study period. The process through which this spread occurred is likely only partially due to local dispersal. The probability of successfully overwintering was likewise low, but both these parameters increased over the study period. This suggests that the presence of Ae. albopictus has been largely due to repeated introductions, but that in recent years populations may have become established and are leading to an increase in locally driven dispersal. There was considerable genetic diversity among populations in Illinois, with 13 distinct haplotypes present in 10 sampling locations, several of which matched haplotypes previously found to be present in locations such as Texas or Japan. Further research is needed to understand how the combination of continued propagule pressure and establishment of populations are driving the increase and expansion of this invasive mosquito along its northern distribution limit.

Documentation of the Expansion of the Gulf Coast Tick (Amblyomma maculatum) and Rickettsia parkeri: First Report in Illinois.

The Gulf Coast tick, Amblyomma maculatum, is of public and veterinary health concern, as it is the primary vector of Rickettsia parkeri and Hepatozoon americanum, causative agents of Rickettsiosis and American canine hepatozoonosis. The Gulf Coast tick's range has expanded over the last 50 yr into the mid-Atlantic states, and its expansion is expected to continue northward. We are reporting the presence of A. maculatum for the first time in Illinois, including a total of 18 specimens collected at 6 different sites during surveys in 2013 and 2019. Fourteen of these specimens were screened for Rickettsia parkeri, which resulted in the detection of this bacteria in 8 samples from 4 counties. By depositing these specimens in scientific collections, we provide materialistic evidence of their establishment in 2 counties. We urge health officials to rely on and use scientific collections to document the expansion of these and other vectors across the country. Additionally, we recommend that health practitioners become aware of the clinical similarities between Rocky Mountain Spotted Fever (caused by Rickettsia rickettsii) and "tidewater" fever (caused by R. parkeri).

A generic arboviral model framework for exploring trade-offs between vector control and environmental concerns.

Effective public health measures must balance potentially conflicting demands from populations they serve. In the case of infectious disease risks from mosquito-borne infections, such as Zika virus, public concern about the pathogen may be counterbalanced by public concern about environmental contamination from chemical agents used for vector control. Here we introduce a generic framework for modeling how the spread of an infectious pathogen might lead to varying public perceptions, and therefore tolerance, of both disease risk and pesticide use. We consider how these dynamics might impact the spread of a vector-borne disease. We tailor and parameterize our model for direct application to Zika virus as spread by Aedes aegypti mosquitoes, though the framework itself has broad applicability to any arboviral infection. We demonstrate how public risk perception of both disease and pesticides may drastically impact the spread of a mosquito-borne disease in a susceptible population. We conclude that models hoping to inform public health decision making about how best to mitigate arboviral disease risks should explicitly consider the potential public demand for, or rejection of, chemical control of mosquito populations.

Contrasting the value of targeted versus area-wide mosquito control scenarios to limit arbovirus transmission with human mobility patterns based on different tropical urban population centers.

Vector control is still our primary intervention for both prevention and mitigation of epidemics of many vector-borne diseases. Efficiently targeting control measures is important since control can involve substantial economic costs. Targeting is not always straightforward, as transmission of vector-borne diseases is affected by various types of host movement. Here we assess how taking daily commuting patterns into consideration can help improve vector control efforts. We examine three tropical urban centers (San Juan, Recife, and Jakarta) that have recently been exposed to Zika and/or dengue infections and consider whether the distribution of human populations and resulting commuting flows affects the optimal scale at which control interventions should be implemented. We developed a stochastic, spatial model and investigated four control scenarios. The scenarios differed in the spatial extent of their implementation and were: 1) a response at the level of an individual neighborhood; 2) a response targeted at a neighborhood in which infected humans were detected and the one with which it was most strongly connected by human movement; 3) a limited area-wide response where all neighborhoods within a certain radius of the focal area were included; and 4) a collective response where all participating neighborhoods implemented control. The relative effectiveness of the scenarios varied only slightly between different settings, with the number of infections averted over time increasing with the scale of implementation. This difference depended on the efficacy of control at the neighborhood level. At low levels of efficacy, the scenarios mirrored each other in infections averted. At high levels of efficacy, impact increased with the scale of the intervention. As a result, the choice between scenarios will not only be a function of the amount of effort decision-makers are willing to invest, but largely epend on the overall effectiveness of vector control approaches.

(Meta)population dynamics determine effective spatial distributions of mosquito-borne disease control.

Recent epidemics of mosquito-borne dengue and Zika viruses demonstrate the urgent need for effective measures to control these diseases. The best method currently available to prevent or reduce the size of outbreaks is to reduce the abundance of their mosquito vectors, but there is little consensus on which mechanisms of control are most effective, or when and where they should be implemented. Although the optimal methods are likely context dependent, broadly applicable strategies for mosquito control, such as how to distribute limited resources across a landscape in times of high epidemic risk, can mitigate (re)emerging outbreaks. We used mathematical simulations to examine how the spatial distribution of larval mosquito control affects the size of disease outbreaks, and how mosquito metapopulation dynamics and demography might impact the efficacy of different spatial distributions of control. We found that the birth rate and mechanism of density-dependent regulation of mosquito populations affected the average outbreak size across all control distributions. These factors also determined whether control distributions favoring the interior or the edges of the landscape most effectively reduced human infections. Thus, understanding local mosquito population regulation and dispersion can lead to more effective control strategies.

The relationship between size and longevity of the malaria vector Anopheles gambiae (s.s.) depends on the larval environment.

BACKGROUND: Understanding the variation in vector-borne disease transmission intensity across time and space relies on a thorough understanding of the impact of environmental factors on vectorial capacity traits of mosquito populations. This is driven primarily by variation in larval development and growth, with carryover effects influencing adult traits such as longevity and adult body size. The relationship between body size and longevity strongly affects the evolution of life histories and the epidemiology of vector-borne diseases. This relationship ranges from positive to negative but the reasons for this variability are not clear. Both traits depend on a number of environmental factors, but primarily on temperature as well as availability of nutritional resources. We therefore asked how the larval environment of the mosquito Anopheles gambiae Giles (sensu stricto) (Diptera: Culicidae) affects the relationship between body size and longevity. METHODS: We reared the larvae of An. gambiae individually at three temperatures (21, 25 and 29 °C) and two food levels (the standard and 50% of our laboratory diet) and measured adult size and longevity. We estimated the direct and indirect (via adult size) effects of food and temperature on longevity with a piecewise structural equation model (SEM). RESULTS: We confirmed the direct effects of food and temperature during larval development on body size, as wing length decreased with increasing temperature and decreasing food levels. While the overall relationship between size and longevity was weak, we measured striking differences among environments. At 25 °C there was no clear relationship between size and longevity; at 29 °C the association was negative with standard food but positive with low food; whereas at 21 °C it was positive with standard food but negative with low food. CONCLUSIONS: The larval environment influences the adult's fitness in complex ways with larger mosquitoes living longer in some environments but not in others. This confirmed our hypothesis that the relationship between size and longevity is not limited to a positive correlation. A better understanding of this relationship and its mechanisms may improve the modelling of the transmission of vector borne diseases, the evolution of life history traits, and the influence of vector control.

The importance of being urgent: The impact of surveillance target and scale on mosquito-borne disease control.

With the emergence or re-emergence of numerous mosquito-borne diseases in recent years, effective methods for emergency vector control responses are necessary to reduce human infections. Current vector control practices often vary significantly between different jurisdictions, and are executed independently and at different spatial scales. Various types of surveillance information (e.g. number of human infections or adult mosquitoes) trigger the implementation of control measures, though the target and scale of surveillance vary locally. This patchy implementation of control measures likely alters the efficacy of control. We modeled six different scenarios, with larval mosquito control occurring in response to surveillance data of different types and at different scales (e.g. across the landscape or in each patch). Our results indicate that: earlier application of larvicide after an escalation of disease risk achieves much greater reductions in human infections than later control implementation; uniform control across the landscape provides better outbreak mitigation than patchy control application; and different types of surveillance data require different levels of sensitivity in their collection to effectively inform control measures. Our simulations also demonstrate a potential logical fallacy of reactive, surveillance-driven vector control: measures stop being implemented as soon as they are deemed effective. This false sense of security leads to patchier control efforts that will do little to curb the size of future vector-borne disease outbreaks. More investment should be placed in collecting high quality information that can trigger early and uniform implementation, while researchers work to discover more informative metrics of human risk to trigger more effective control.

Human movement, cooperation and the effectiveness of coordinated vector control strategies.

Vector-borne disease transmission is often typified by highly focal transmission and influenced by movement of hosts and vectors across different scales. The ecological and environmental conditions (including those created by humans through vector control programmes) that result in metapopulation dynamics remain poorly understood. The development of control strategies that would most effectively limit outbreaks given such dynamics is particularly urgent given the recent epidemics of dengue, chikungunya and Zika viruses. We developed a stochastic, spatial model of vector-borne disease transmission, allowing for movement of hosts between patches. Our model is applicable to arbovirus transmission by Aedes aegypti in urban settings and was parametrized to capture Zika virus transmission in particular. Using simulations, we investigated the extent to which two aspects of vector control strategies are affected by human commuting patterns: the extent of coordination and cooperation between neighbouring communities. We find that transmission intensity is highest at intermediate levels of host movement. The extent to which coordination of control activities among neighbouring patches decreases the prevalence of infection is affected by both how frequently humans commute and the proportion of neighbouring patches that commits to vector surveillance and control activities. At high levels of host movement, patches that do not contribute to vector control may act as sources of infection in the landscape, yet have comparable levels of prevalence as patches that do cooperate. This result suggests that real cooperation among neighbours will be critical to the development of effective pro-active strategies for vector-borne disease control in today's commuter-linked communities.

Implications of Heterogeneous Biting Exposure and Animal Hosts on Trypanosomiasis brucei gambiense Transmission and Control.

The gambiense form of sleeping sickness is a neglected tropical disease, which is presumed to be anthroponotic. However, the parasite persists in human populations at levels of considerable rarity and as such the existence of animal reservoirs has been posited. Clarifying the impact of animal host reservoirs on the feasibility of interrupting sleeping sickness transmission through interventions is a matter of urgency. We developed a mathematical model allowing for heterogeneous exposure of humans to tsetse, with animal populations that differed in their ability to transmit infections, to investigate the effectiveness of two established techniques, screening and treatment of at-risk populations, and vector control. Importantly, under both assumptions, an integrated approach of human screening and vector control was supported in high transmission areas. However, increasing the intensity of vector control was more likely to eliminate transmission, while increasing the intensity of human screening reduced the time to elimination. Non-human animal hosts played important, but different roles in HAT transmission, depending on whether or not they contributed as reservoirs. If they did not serve as reservoirs, sensitivity analyses suggested their attractiveness may instead function as a sink for tsetse bites. These outcomes highlight the importance of understanding the ecological and environmental context of sleeping sickness in optimizing integrated interventions, particularly for moderate and low transmission intensity settings.

Mathematical models of human african trypanosomiasis epidemiology.

Human African trypanosomiasis (HAT), commonly called sleeping sickness, is caused by Trypanosoma spp. and transmitted by tsetse flies (Glossina spp.). HAT is usually fatal if untreated and transmission occurs in foci across sub-Saharan Africa. Mathematical modelling of HAT began in the 1980s with extensions of the Ross-Macdonald malaria model and has since consisted, with a few exceptions, of similar deterministic compartmental models. These models have captured the main features of HAT epidemiology and provided insight on the effectiveness of the two main control interventions (treatment of humans and tsetse fly control) in eliminating transmission. However, most existing models have overestimated prevalence of infection and ignored transient dynamics. There is a need for properly validated models, evolving with improved data collection, that can provide quantitative predictions to help guide control and elimination strategies for HAT.

Effects of bed net use, female size, and plant abundance on the first meal choice (blood vs sugar) of the malaria mosquito Anopheles gambiae.

BACKGROUND: The purpose of this study was to determine whether the sugar-or-blood meal choice of Anopheles gambiae females one day after emergence is influenced by blood-host presence and accessibility, nectariferous plant abundance, and female size. This tested the hypothesis that the initial meal of female An. gambiae is sugar, even when a blood host is available throughout the night, and, if not, whether the use of a bed net diverts mosquitoes to sugar sources. METHODS: Females and males <1-day post-emergence were released in a mesocosm. Overnight they had access to either one or six Senna didymobotrya plants. Simultaneously they had access to a human blood host, either for 8 h or for only 30 min at dusk and dawn (the remainder of the night being excluded by an untreated bed net). In a third situation, the blood host was not present. All mosquitoes were collected in the morning. Their wing lengths, an indicator of pre-meal energetic state, were measured, and their meal choice was determined by the presence of midgut blood and of fructose. RESULTS: Female sugar feeding after emergence was facultative. When a blood host was accessible for 8 h per night, 92% contained blood, and only 3.7% contained sugar. Even with the use of a bed net, 78% managed to obtain a blood meal during the 30 min of accessibility at dusk or dawn, but 14% of females were now fructose-positive. In the absence of a blood host, and when either one or six plants were available, a total of 21.7% and 23.6% of females and 30.8% and 43.5% of males contained fructose, respectively. Feeding on both sugar and blood was more likely with bed net use and with greater plant abundance. Further, mosquitoes that fed on both resources were more often small and had taken a sugar meal earlier than the blood meal. The abundance of sugar hosts also affected the probability of sugar feeding by males and the amount of fructose obtained by both males and females. CONCLUSION: Even in an abundance of potential sugar sources, female An. gambiae appear to prefer a nearby human source of blood. However, the decision to take sugar was more likely if energy reserves were low. Results probably would differ if sugar hosts were more attractive or yielded larger sugar meals. The diversion of energetically deprived mosquitoes to sugar sources suggests a possible synergy between bed nets and sugar-based control methods.