Ingested histamine and serotonin interact to alter Anopheles stephensi feeding and flight behavior and infection with Plasmodium parasites.

Blood levels of histamine and serotonin (5-HT) are altered in human malaria, and, at these levels, we have shown they have broad, independent effects on Anopheles stephensi following ingestion by this invasive mosquito. Given that histamine and 5-HT are ingested together under natural conditions and that histaminergic and serotonergic signaling are networked in other organisms, we examined effects of combinations of these biogenic amines provisioned to A. stephensi at healthy human levels (high 5-HT, low histamine) or levels associated with severe malaria (low 5-HT, high histamine). Treatments were delivered in water (priming) before feeding A. stephensi on Plasmodium yoelii-infected mice or via artificial blood meal. Relative to effects of histamine and 5-HT alone, effects of biogenic amine combinations were complex. Biogenic amine treatments had the greatest impact on the first oviposition cycle, with high histamine moderating low 5-HT effects in combination. In contrast, clutch sizes were similar across combination and individual treatments. While high histamine alone increased uninfected A. stephensi weekly lifetime blood feeding, neither combination altered this tendency relative to controls. The tendency to re-feed 2 weeks after the first blood meal was altered by combination treatments, but this depended on mode of delivery. For blood delivery, malaria-associated treatments yielded higher percentages of fed females relative to healthy-associated treatments, but the converse was true for priming. Female mosquitoes treated with the malaria-associated combination exhibited enhanced flight behavior and object inspection relative to controls and healthy combination treatment. Mosquitoes primed with the malaria-associated combination exhibited higher mean oocysts and sporozoite infection prevalence relative to the healthy combination, with high histamine having a dominant effect on these patterns. Compared with uninfected A. stephensi, the tendency of infected mosquitoes to take a second blood meal revealed an interaction of biogenic amines with infection. We used a mathematical model to project the impacts of different levels of biogenic amines and associated changes on outbreaks in human populations. While not all outbreak parameters were impacted the same, the sum of effects suggests that histamine and 5-HT alter the likelihood of transmission by mosquitoes that feed on hosts with symptomatic malaria versus a healthy host.

Spatiotemporal and meteorological relationships in dengue transmission in the Dominican Republic, 2015-2019.

Dengue has broadened its global distribution substantially in the past two decades, and many endemic areas are experiencing increases in incidence. The Dominican Republic recently experienced its two largest outbreaks to date with 16,836 reported cases in 2015 and 20,123 reported cases in 2019. With continued increases in dengue transmission, developing tools to better prepare healthcare systems and mosquito control agencies is of critical importance. Before such tools can be developed, however, we must first better understand potential drivers of dengue transmission. To that end, we focus in this paper on determining relationships between climate variables and dengue transmission with an emphasis on eight provinces and the capital city of the Dominican Republic in the period 2015-2019. We present summary statistics for dengue cases, temperature, precipitation, and relative humidity in this period, and we conduct an analysis of correlated lags between climate variables and dengue cases as well as correlated lags among dengue cases in each of the nine locations. We find that the southwestern province of Barahona had the largest dengue incidence in both 2015 and 2019. Among all climate variables considered, lags between relative humidity variables and dengue cases were the most frequently correlated. We found that most locations had significant correlations with cases in other locations at lags of zero weeks. These results can be used to improve predictive models of dengue transmission in the country.

Relationship between Climate Variables and Dengue Incidence in Argentina.

BACKGROUND: Climate change is an important driver of the increased spread of dengue from tropical and subtropical regions to temperate areas around the world. Climate variables such as temperature and precipitation influence the dengue vector's biology, physiology, abundance, and life cycle. Thus, an analysis is needed of changes in climate change and their possible relationships with dengue incidence and the growing occurrence of epidemics recorded in recent decades. OBJECTIVES: This study aimed to assess the increasing incidence of dengue driven by climate change at the southern limits of dengue virus transmission in South America. METHODS: We analyzed the evolution of climatological, epidemiological, and biological variables by comparing a period of time without the presence of dengue cases (1976-1997) to a more recent period of time in which dengue cases and important outbreaks occurred (1998-2020). In our analysis, we consider climate variables associated with temperature and precipitation, epidemiological variables such as the number of reported dengue cases and incidence of dengue, and biological variables such as the optimal temperature ranges for transmission of dengue vector. RESULTS: The presence of dengue cases and epidemic outbreaks are observed to be consistent with positive trends in temperature and anomalies from long-term means. Dengue cases do not seem to be associated with precipitation trends and anomalies. The number of days with optimal temperatures for dengue transmission increased from the period without dengue cases to the period with occurrences of dengue cases. The number of months with optimal transmission temperatures also increased between periods but to a lesser extent. CONCLUSIONS: The higher incidence of dengue virus and its expansion to different regions of Argentina seem to be associated with temperature increases in the country during the past two decades. The active surveillance of both the vector and associated arboviruses, together with continued meteorological data collection, will facilitate the assessment and prediction of future epidemics that use trends in the accelerated changes in climate. Such surveillance should go hand in hand with efforts to improve the understanding of the mechanisms driving the geographic expansion of dengue and other arboviruses beyond the current limits. https://doi.org/10.1289/EHP11616.

Modeling the effects of Aedes aegypti's larval environment on adult body mass at emergence.

Mosquitoes vector harmful pathogens that infect millions of people every year, and developing approaches to effectively control mosquitoes is a topic of great interest. However, the success of many control measures is highly dependent upon ecological, physiological, and life history traits of mosquito species. The behavior of mosquitoes and their potential to vector pathogens can also be impacted by these traits. One trait of interest is mosquito body mass, which depends upon many factors associated with the environment in which juvenile mosquitoes develop. Our experiments examined the impact of larval density on the body mass of Aedes aegypti mosquitoes, which are important vectors of dengue, Zika, yellow fever, and other pathogens. To investigate the interactions between the larval environment and mosquito body mass, we built a discrete time mathematical model that incorporates body mass, larval density, and food availability and fit the model to our experimental data. We considered three categories of model complexity informed by data, and selected the best model within each category using Akaike's Information Criterion. We found that the larval environment is an important determinant of the body mass of mosquitoes upon emergence. Furthermore, we found that larval density has greater impact on body mass of adults at emergence than on development time, and that inclusion of density dependence in the survival of female aquatic stages in models is important. We discuss the implications of our results for the control of Aedes mosquitoes and on their potential to spread disease.

Observational Characterization of the Ecological and Environmental Features Associated with the Presence of Oropouche Virus and the Primary Vector Culicoides paraenesis: Data Synthesis and Systematic Review.

Oropouche virus (OROV), a member of the Orthobunyavirus genus, is an arthropod-borne virus (arbovirus) and is the etiologic agent of human and animal disease. The primary vector of OROV is presumed to be the biting midge, Culicoides paraenesis, though Culex quinquefasciatus, Cq. venezuelensis, and Aedes serratus mosquitoes are considered secondary vectors. The objective of this systematic review is to characterize locations where OROV and/or its primary vector have been detected. Synthesis of known data through review of published literature regarding OROV and vectors was carried out through two independent searches: one search targeted to OROV, and another targeted towards the primary vector. A total of 911 records were returned, but only 90 (9.9%) articles satisfied all inclusion criteria. When locations were characterized, some common features were noted more frequently than others, though no one characteristic was significantly associated with presence of OROV using a logistic classification model. In a separate correlation analysis, vector presence was significantly correlated only with the presence of restingas. The lack of significant relationships is likely due to the paucity of data regarding OROV and its eco-epidemiology and highlights the importance of continued focus on characterizing this and other neglected tropical diseases.

Dengue emergence in the temperate Argentinian province of Santa Fe, 2009-2020.

Dengue virus (DENV) transmission occurs primarily in tropical and subtropical climates, but within the last decade it has extended to temperate regions. Santa Fe, a temperate province in Argentina, has experienced an increase in dengue cases and virus circulation since 2009, with the recent 2020 outbreak being the largest in the province to date. The aim of this work is to describe spatio-temporal fluctuations of dengue cases from 2009 to 2020 in Santa Fe Province. The data presented in this work provide a detailed description of DENV transmission for Santa Fe Province by department. These data are useful to assist in investigating drivers of dengue emergence in Santa Fe Province and for developing a better understanding of the drivers and the impacts of ongoing dengue emergence in temperate regions across the world. This work provides data useful for future studies including those investigating socio-ecological, climatic, and environmental factors associated with DENV transmission, as well as those investigating other variables related to the biology and the ecology of vector-borne diseases.

Understanding the role of temporal variation of environmental variables in predicting Aedes aegypti oviposition activity in a temperate region of Argentina.

Environmental variables related to vegetation and weather are some of the most influential factors that impacting Aedes (Stegomya) aegypti, a mosquito vector of dengue, chikungunya and Zika viruses. In this paper, we aim to develop temporal predictive models for Ae. aegypti oviposition activity utilizing vegetation and meteorological variables as predictors in Córdoba city (Argentina). Eggs were collected using ovitraps placed throughout the city from 2009 to 2012 that were replaced weekly. Temporal generalized linear mixed models were developed with negative binomial distributions of errors that model average number of eggs collected weekly as a function of vegetation and meteorological variables with time lags. The best model included a vegetation index, vapor pressure of water, precipitation and photoperiod. With each unit of increment in vegetation index per week the average number of eggs increased by 1.71 in the third week. Furthermore, each millimeter increase of accumulated rain during 4 weeks was associated with a decrease of 0.668 in the average number of eggs found in the following week. This negative effect of precipitation could occur during abundant rainfalls that fill containers completely, thereby depriving females of oviposition sites and leading them to search for other suitable breeding sites. Furthermore, the average number of eggs increased with the photoperiod at low values of mean vapor pressure; however the average number of eggs decreased at high values of mean vapor pressure, and the positive relationship between the response variable and mean vapor pressure was stronger at low values of photoperiod. Additionally, minimum temperature was associated positively with oviposition activity and that low minimum temperatures could be a limiting factor in Ae. aegypti oviposition activity. Our results emphasize the important role that climatic variables such as temperature, precipitation, and vapor pressure play in Ae. aegypti oviposition activity and how these variables along with vegetation indices can be used to inform predictive temporal models of Ae. aegypti population dynamics that can be used for informing mosquito population control and arbovirus mitigation strategies.

Climate change and viral emergence: evidence from Aedes-borne arboviruses.

Climate change is leading to increases in global temperatures and erratic precipitation patterns, both of which are contributing to the expansion of mosquito-borne arboviruses and the populations of the mosquitos that vector them. Herein, we review recent evidence of emergence and expansion of arboviruses transmitted by Aedes mosquitos that has been driven in part by environmental changes. We present as a case study of recent work from Córdoba, Argentina, where dengue has been actively emerging in the past decade. We review recent empirical and modeling studies that aim to understand the impact of climate on future expansion of arboviruses, and we highlight gaps in empirical studies linking climate to arbovirus transmission at regional levels.

The impact of local population genetic background on the spread of the selfish element Medea-1 in red flour beetles.

Selfish genetic elements have been found in the genomes of many species, yet our understanding of their evolutionary dynamics is only partially understood. A number of distinct selfish Medea elements are naturally present in many populations of the red flour beetle (Tribolium castaneum). Although these Medea elements are predicted by models to increase in frequency within populations because any offspring of a Medea-bearing mother that do not inherit at least one Medea allele will die, experiments demonstrating an increase in a naturally occurring Medea element are lacking. Our survey of the specific Medea element, M1, in the United States showed that it had a patchy geographic distribution. From the survey, it could not be determined if this distribution was caused by a slow process of M1 colonization of discrete populations or if some populations lacked M1 because they had genetic factors conferring resistance to the Medea mechanism. We show that populations with naturally low to intermediate M1 frequencies likely represent transient states during the process of Medea spread. Furthermore, we find no evidence that genetic factors are excluding M1 from US populations where the element is not presently found. We also show how a known suppressor of Medea can impair the increase of M1 in populations and discuss the implications of our findings for pest-management applications of Medea elements.

Arbovirus emergence in the temperate city of Córdoba, Argentina, 2009-2018.

The distribution of arbovirus disease transmission is expanding from the tropics and subtropics into temperate regions worldwide. The temperate city of Córdoba, Argentina has been experiencing the emergence of dengue virus, transmitted by the mosquito Aedes aegypti, since 2009, when autochthonous transmission of the virus was first recorded in the city. The aim of this work is to characterize the emergence of dengue and related arboviruses (Zika and chikungunya) in Córdoba since 2009. Herein, we present a data set with all known information about transmission of dengue, Zika, and chikungunya viruses in Córdoba, Argentina from 2009-2018, including what information is known of dengue virus (DENV) serotypes in circulation and origins of imported cases. The data presented in this work will assist researchers in investigating drivers of arbovirus emergence and transmission in Córdoba, Argentina and contribute to a better understanding of the global problem of the expanding distribution of arbovirus disease transmission.

Temperature impacts on dengue emergence in the United States: Investigating the role of seasonality and climate change.

Tropical mosquito-borne viruses have been expanding into more temperate regions in recent decades. This is partly due to the coupled effects of temperature on mosquito life history traits and viral infection dynamics and warming surface temperatures, resulting in more suitable conditions for vectors and virus transmission. In this study, we use a deterministic ordinary differential equations model to investigate how seasonal and diurnal temperature fluctuations affect the potential for dengue transmission in six U.S. cities. We specifically consider temperature-dependent mosquito larval development, adult mosquito mortality, and the extrinsic incubation period of the virus. We show that the ability of introductions to lead to outbreaks depends upon the relationship between a city's temperature profile and the time of year at which the initial case is introduced. We also investigate how the potential for outbreaks changes with predicted future increases in mean temperatures due to climate change. We find that climate change will likely lead to increases in suitability for dengue transmission and will increase the periods of the year in which introductions may lead to outbreaks, particularly in cities that typically have mild winters and warm summers, such as New Orleans, Louisiana, and El Paso, Texas. We discuss our results in the context of temperature heterogeneity within and across cities and how these differences may impact the potential for dengue emergence given present day and predicted future temperatures.

Modeling Mosquito-Borne Disease Spread in U.S. Urbanized Areas: The Case of Dengue in Miami.

Expansion of mosquito-borne pathogens into more temperate regions of the world necessitates tools such as mathematical models for understanding the factors that contribute to the introduction and emergence of a disease in populations naïve to the disease. Often, these models are not developed and analyzed until after a pathogen is detected in a population. In this study, we develop a spatially explicit stochastic model parameterized with publicly available U.S. Census data for studying the potential for disease spread in Urbanized Areas of the United States. To illustrate the utility of the model, we specifically study the potential for introductions of dengue to lead to autochthonous transmission and outbreaks in a population representative of the Miami Urbanized Area, where introductions of dengue have occurred frequently in recent years. We describe seasonal fluctuations in mosquito populations by fitting a population model to trap data provided by the Miami-Dade Mosquito Control Division. We show that the timing and location of introduced cases could play an important role in determining both the probability that local transmission occurs as well as the total number of cases throughout the entire region following introduction. We show that at low rates of clinical presentation, small outbreaks of dengue could go completely undetected during a season, which may confound mitigation efforts that rely upon detection. We discuss the sensitivity of the model to several critical parameter values that are currently poorly characterized and motivate the collection of additional data to strengthen the predictive power of this and similar models. Finally, we emphasize the utility of the general structure of this model in studying mosquito-borne diseases such as chikungunya and Zika virus in other regions.

Dengue and chikungunya: modelling the expansion of mosquito-borne viruses into naïve populations.

With the recent global spread of a number of mosquito-borne viruses, there is an urgent need to understand the factors that contribute to the ability of viruses to expand into naïve populations. Using dengue and chikungunya viruses as case studies, we detail the necessary components of the expansion process: presence of the mosquito vector; introduction of the virus; and suitable conditions for local transmission. For each component we review the existing modelling approaches that have been used to understand recent emergence events or to assess the risk of future expansions. We identify gaps in our knowledge that are related to each of the distinct aspects of the human-mosquito transmission cycle: mosquito ecology; human-mosquito contact; mosquito-virus interactions; and human-virus interactions. Bridging these gaps poses challenges to both modellers and empiricists, but only through further integration of models and data will we improve our ability to better understand, and ultimately control, several infectious diseases that exert a significant burden on human health.

Feasible introgression of an anti-pathogen transgene into an urban mosquito population without using gene-drive.

BACKGROUND: Introgressing anti-pathogen constructs into wild vector populations could reduce disease transmission. It is generally assumed that such introgression would require linking an anti-pathogen gene with a selfish genetic element or similar technologies. Yet none of the proposed transgenic anti-pathogen gene-drive mechanisms are likely to be implemented as public health measures in the near future. Thus, much attention now focuses instead on transgenic strategies aimed at mosquito population suppression, an approach generally perceived to be practical. By contrast, aiming to replace vector competent mosquito populations with vector incompetent populations by releasing mosquitoes carrying a single anti-pathogen gene without a gene-drive mechanism is widely considered impractical. METHODOLOGY/PRINCIPAL FINDINGS: Here we use Skeeter Buster, a previously published stochastic, spatially explicit model of Aedes aegypti to investigate whether a number of approaches for releasing mosquitoes with only an anti-pathogen construct would be efficient and effective in the tropical city of Iquitos, Peru. To assess the performance of such releases using realistic release numbers, we compare the transient and long-term effects of this strategy with two other genetic control strategies that have been developed in Ae. aegypti: release of a strain with female-specific lethality, and a strain with both female-specific lethality and an anti-pathogen gene. We find that releasing mosquitoes carrying only an anti-pathogen construct can substantially decrease vector competence of a natural population, even at release ratios well below that required for the two currently feasible alternatives that rely on population reduction. Finally, although current genetic control strategies based on population reduction are compromised by immigration of wild-type mosquitoes, releasing mosquitoes carrying only an anti-pathogen gene is considerably more robust to such immigration. CONCLUSIONS/SIGNIFICANCE: Contrary to the widely held view that transgenic control programs aimed at population replacement require linking an anti-pathogen gene to selfish genetic elements, we find releasing mosquitoes in numbers much smaller than those considered necessary for transgenic population reduction can result in comparatively rapid and robust population replacement. In light of this non-intuitive result, directing efforts to improve rearing capacity and logistical support for implementing releases, and reducing the fitness costs of existing recombinant technologies, may provide a viable, alternative route to introgressing anti-pathogen transgenes under field conditions.

Antipathogen genes and the replacement of disease-vectoring mosquito populations: a model-based evaluation.

Recently, genetic strategies aimed at controlling populations of disease-vectoring mosquitoes have received considerable attention as alternatives to traditional measures. Theoretical studies have shown that female-killing (FK), antipathogen (AP), and reduce and replace (R&R) strategies can each decrease the number competent vectors. In this study, we utilize a mathematical model to evaluate impacts on competent Aedes aegypti populations of FK, AP, and R&R releases as well as hybrid strategies that result from combinations of these three approaches. We show that while the ordering of efficacy of these strategies depends upon population life history parameters, sex ratio of releases, and switch time in combination strategies, AP-only and R&R/AP releases typically lead to the greatest long-term reduction in competent vectors. R&R-only releases are often less effective at long-term reduction of competent vectors than AP-only releases or R&R/AP releases. Furthermore, the reduction in competent vectors caused by AP-only releases is easier to maintain than that caused by FK-only or R&R-only releases even when the AP gene confers a fitness cost. We discuss the roles that density dependence and inclusion of females play in the order of efficacy of the strategies. We anticipate that our results will provide added impetus to continue developing AP strategies.

A reduce and replace strategy for suppressing vector-borne diseases: insights from a stochastic, spatial model.

Two basic strategies have been proposed for using transgenic Aedes aegypti mosquitoes to decrease dengue virus transmission: population reduction and population replacement. Here we model releases of a strain of Ae. aegypti carrying both a gene causing conditional adult female mortality and a gene blocking virus transmission into a wild population to assess whether such releases could reduce the number of competent vectors. We find this "reduce and replace" strategy can decrease the frequency of competent vectors below 50% two years after releases end. Therefore, this combined approach appears preferable to releasing a strain carrying only a female-killing gene, which is likely to merely result in temporary population suppression. However, the fixation of anti-pathogen genes in the population is unlikely. Genetic drift at small population sizes and the spatially heterogeneous nature of the population recovery after releases end prevent complete replacement of the competent vector population. Furthermore, releasing more individuals can be counter-productive in the face of immigration by wild-type mosquitoes, as greater population reduction amplifies the impact wild-type migrants have on the long-term frequency of the anti-pathogen gene. We expect the results presented here to give pause to expectations for driving an anti-pathogen construct to fixation by relying on releasing individuals carrying this two-gene construct. Nevertheless, in some dengue-endemic environments, a spatially heterogeneous decrease in competent vectors may still facilitate decreasing disease incidence.

A reduce and replace strategy for suppressing vector-borne diseases: insights from a deterministic model.

Genetic approaches for controlling disease vectors have aimed either to reduce wild-type populations or to replace wild-type populations with insects that cannot transmit pathogens. Here, we propose a Reduce and Replace (R&R) strategy in which released insects have both female-killing and anti-pathogen genes. We develop a mathematical model to numerically explore release strategies involving an R&R strain of the dengue vector Aedes aegypti. We show that repeated R&R releases may lead to a temporary decrease in mosquito population density and, in the absence of fitness costs associated with the anti-pathogen gene, a long-term decrease in competent vector population density. We find that R&R releases more rapidly reduce the transient and long-term competent vector densities than female-killing releases alone. We show that releases including R&R females lead to greater reduction in competent vector density than male-only releases. The magnitude of reduction in total and competent vectors depends upon the release ratio, release duration, and whether females are included in releases. Even when the anti-pathogen allele has a fitness cost, R&R releases lead to greater reduction in competent vectors than female-killing releases during the release period; however, continued releases are needed to maintain low density of competent vectors long-term. We discuss the results of the model as motivation for more detailed studies of R&R strategies.

Field cage studies and progressive evaluation of genetically-engineered mosquitoes.

BACKGROUND: A genetically-engineered strain of the dengue mosquito vector Aedes aegypti, designated OX3604C, was evaluated in large outdoor cage trials for its potential to improve dengue prevention efforts by inducing population suppression. OX3604C is engineered with a repressible genetic construct that causes a female-specific flightless phenotype. Wild-type females that mate with homozygous OX3604C males will not produce reproductive female offspring. Weekly introductions of OX3604C males eliminated all three targeted Ae. aegypti populations after 10-20 weeks in a previous laboratory cage experiment. As part of the phased, progressive evaluation of this technology, we carried out an assessment in large outdoor field enclosures in dengue endemic southern Mexico. METHODOLOGY/PRINCIPAL FINDINGS: OX3604C males were introduced weekly into field cages containing stable target populations, initially at 10:1 ratios. Statistically significant target population decreases were detected in 4 of 5 treatment cages after 17 weeks, but none of the treatment populations were eliminated. Mating competitiveness experiments, carried out to explore the discrepancy between lab and field cage results revealed a maximum mating disadvantage of up 59.1% for OX3604C males, which accounted for a significant part of the 97% fitness cost predicted by a mathematical model to be necessary to produce the field cage results. CONCLUSIONS/SIGNIFICANCE: Our results indicate that OX3604C may not be effective in large-scale releases. A strain with the same transgene that is not encumbered by a large mating disadvantage, however, could have improved prospects for dengue prevention. Insights from large outdoor cage experiments may provide an important part of the progressive, stepwise evaluation of genetically-engineered mosquitoes.

Mathematical models as aids for design and development of experiments: the case of transgenic mosquitoes.

We demonstrate the utility of models as aids in the design and development of experiments aimed at measuring the effects of proposed vector population control strategies. We describe the exploration of a stochastic, age-structured model that simulates field cage experiments that test the ability of a female-killing strain of the mosquito Aedes aegypti (L.) to suppress a wild-type population. Model output predicts that choices of release ratio and population size can impact mean extinction time and variability in extinction time among experiments. We find that unless fitness costs are >80% they will not be detectable in experiments with high release ratios. At lower release ratios, the predicted length of the experiment increases significantly for fitness costs >20%. Experiments with small populations may more accurately reflect field conditions, but extinction can occur even in the absence of a functional female-killing construct because of stochastic effects. We illustrate how the model can be used to explore experimental designs that aim to study the impact of density dependence and immigration; predictions indicate that cage population eradication may not always be obtainable in an operationally realistic time frame. We propose a method to predict the extinction time of a cage population based on the rate of population reduction with the goal of shortening the duration of the experiment. We discuss the model as a tool for exploring and assessing the utility of a wider range of scenarios than would be feasible to test experimentally because of financial and temporal restraints.