1. Computational and Systems Biology
  2. Ecology
Download icon

Data-driven identification of potential Zika virus vectors

  1. Michelle V Evans Is a corresponding author
  2. Tad A Dallas
  3. Barbara A Han
  4. Courtney C Murdock
  5. John M Drake
  1. University of Georgia, United States
  2. Center for the Ecology of Infectious Diseases, University of Georgia, United States
  3. University of California-Davis, United States
  4. Cary Institute of Ecosystem Studies, United States
  5. Center for Tropical Emerging Global Diseases, University of Georgia, United States
  6. Center for Vaccines and Immunology, University of Georgia, United States
  7. River Basin Center, University of Georgia, United States
Research Article
Cited
0
Views
2,019
Comments
0
Cite as: eLife 2017;6:e22053 doi: 10.7554/eLife.22053

Abstract

Zika is an emerging virus whose rapid spread is of great public health concern. Knowledge about transmission remains incomplete, especially concerning potential transmission in geographic areas in which it has not yet been introduced. To identify unknown vectors of Zika, we developed a data-driven model linking vector species and the Zika virus via vector-virus trait combinations that confer a propensity toward associations in an ecological network connecting flaviviruses and their mosquito vectors. Our model predicts that thirty-five species may be able to transmit the virus, seven of which are found in the continental United States, including Culex quinquefasciatus and Cx. pipiens. We suggest that empirical studies prioritize these species to confirm predictions of vector competence, enabling the correct identification of populations at risk for transmission within the United States.

https://doi.org/10.7554/eLife.22053.001

eLife digest

Mosquitoes carry several diseases that pose an emerging threat to society. Outbreaks of these diseases are often sudden and can spread to previously unaffected areas. For example, the Zika virus was discovered in 1947, but only received international attention when it spread to the Americas in 2014, where it caused over 100,000 cases in Brazil alone. While we now recognize the threat Zika can pose for public health, our knowledge about the ecology of the disease remains poor. Nine species of mosquitoes are known to be able to carry the Zika virus, but it cannot be ruled out that other mosquitoes may also be able to spread the disease.

There are hundreds of species of mosquitoes, and testing all of them is difficult and costly. So far, only a small number of species have been tested to see if they transmit Zika. However, computational tools called decision trees could help by predicting which mosquitoes can transmit a virus based on common traits, such as a mosquito's geographic range, or the symptoms of a virus.

Evans et al. used decision trees to create a model that predicts which species of mosquitoes are potential carriers of Zika virus and should therefore be prioritized for testing. The model took into account all known viruses that belong to the same family as Zika virus and the mosquitoes that carry them. Evans et al. predict that 35 species may be able to carry the Zika virus, seven of which are found in the United States. Two of these mosquito species are known to transmit West Nile Virus and are therefore prime examples of species that should be prioritized for testing. Together, the ranges of the seven American species encompass the whole United States, suggesting Zika virus could affect a much larger area than previously anticipated.

The next step following on from this work will be to carry out experiments to test if the 35 mosquitoes identified by the model are actually able to transmit the Zika virus.

https://doi.org/10.7554/eLife.22053.002

Introduction

In 2014, Zika virus was introduced into Brazil and Haiti, from where it rapidly spread throughout the Americas. By January 2017, over 100,000 cases had been confirmed in 24 different states in Brazil (http://ais.paho.org/phip/viz/ed_zika_cases.asp), with large numbers of reports from many other counties in South and Central America (Faria et al., 2016). Originally isolated in Uganda in 1947, the virus remained poorly understood until it began to spread within the South Pacific, including an outbreak affecting 75% of the residents on the island of Yap in 2007 (49 confirmed cases) and over 32,000 cases in the rest of Oceania in 2013–2014, the largest outbreak prior to the Americas (2016-present) (Cao-Lormeau et al., 2016; Duffy et al., 2009). Guillian-Barré syndrome, a neurological pathology associated with Zika virus infection, was first recognized at this time (Cao-Lormeau et al., 2016). Similarly, an increase in newborn microcephaly was found to be correlated with the increase in Zika cases in Brazil in 2015 and 2016 (Schuler-Faccini et al., 2016). For this reason, in February 2016, the World Health Organization declared the American Zika virus epidemic to be a Public Health Emergency of International Concern.

Despite its public health importance, the ecology of Zika virus transmission has been poorly understood until recently. It has been presumed that Aedes aegypti and Ae. albopictus are the primary vectors due to epidemiologic association with Zika virus (Messina et al., 2016), viral isolation from and transmission experiments with field populations (especially in Ae. aegypti [Haddow et al., 2012; Boorman and Porterfield, 1956; Haddow et al., 1964]), and association with related arboviruses (e.g. dengue fever virus, yellow fever virus). Predictions of the potential geographic range of Zika virus in the United States,and associated estimates for the size of the vulnerable population, are therefore primarily based on the distributions of Ae. aegypti and Ae. albopictus, which jointly extend across the Southwest, Gulf coast, and mid-Atlantic regions of the United States (Centers for Disease Control and Prevention, 2016). We reasoned, however, that if other, presently unidentified Zika-competent mosquitoes exist in the Americas, then these projections may be too restricted and therefore optimistically biased. Additionally, recent experimental studies show that the ability of Ae. aegypti and Ae. albopictus to transmit the virus varies significantly across mosquito populations and geographic regions (Chouin-Carneiro et al., 2016), with some populations exhibiting low dissemination rates even though the initial viral titer after inoculation may be high (Diagne et al., 2015). This suggests that in some locations other species may be involved in transmission. The outbreak on Yap, for example, was driven by a different species, Ae. hensilli (Ledermann et al., 2014). Closely related viruses of the Flaviviridae family are vectored by over nine mosquito species, on average (see Supplementary Data). Thus, because Zika virus may be associated with multiple mosquito species, we considered it necessary to develop a more comprehensive list of potential Zika vectors.

The gold standard for identifying competent disease vectors requires isolating virus from field-collected mosquitoes, followed by experimental inoculation and laboratory investigation of viral dissemination throughout the body and to the salivary glands (Barnett, 1960; Hardy et al., 1983), and, when possible, successful transmission back to the vertebrate host (e.g. Komar et al., 2003). Unfortunately, these methods are costly, often underestimate the risk of transmission (Bustamante and Lord, 2010), and the amount of time required for analyses can delay decision making during an outbreak (Day, 2001). To address the problem of identifying potential vector candidates in an actionable time frame, we therefore pursued a data-driven approach to identifying candidate vectors aided by machine learning algorithms for identifying patterns in high dimensional data. If the propensity of mosquito species to associate with Zika virus is statistically associated with common mosquito traits, it is possible to rank mosquito species by the degree of risk represented by their traits – a comparative approach similar to the analysis of risk factors in epidemiology. For instance, a model could be constructed to estimate the statistical discrepancy between the traits of known vectors (i.e., Ae. aegypti, Ae. albopictus, and Ae. hensilli) and the traits of all possible vectors. Unfortunately, this simplistic approach would inevitably fail due to the small amount of available data (i.e., sample size of 3). Thus, we developed an indirect approach that leverages the information contained in the associations among many virus-mosquito pairs to inform us about specific associations. Specifically, our method identifies covariates associated with the propensity for mosquito species to vector any flavivirus. From this, we constructed a model of the mosquito-flavivirus network and then extracted from this model the life history profile and species list of mosquitoes predicted to associate with Zika virus, which we recommend be experimentally tested for Zika virus competence.

Results

In total, we identified 132 vector-virus pairs, consisting of 77 mosquito species and 37 flaviviruses. The majority of these species were Aedes (32) or Culex (24) species. Our supplementary dataset consisted of an additional 103 mosquito species suspected to transmit flaviviruses, but for which evidence of a full transmission cycle does not exist. This resulted in 180 potential mosquito-Zika pairs on which to predict with our trained model. As expected, closely related viruses, such as the four strains of dengue, shared many of the same vectors and were clustered in our network diagram (Figure 1). The distribution of vectors to viruses was uneven, with a few viruses vectored by many mosquito species, and rarer viruses vectored by only one or two species. The virus with the most known competent vectors was West Nile virus (31 mosquito vectors), followed by yellow fever virus (24 mosquito vectors). In general, encephalitic viruses such as West Nile virus were found to be more commonly vectored by Culex mosquitoes and hemorrhagic viruses were found to be more commonly vectored by Aedes mosquitoes (see Gould and Solomon (2008) for further distinctions within Flaviviridae) (Figure 1).

A network diagram of mosquito vectors (circles) and their flavivirus pairs (rectangles).

The Culex mosquitoes (light blue) and primarily encephalitic viruses (blue) are more clustered than the Aedes (orange) and hemmorhagic viruses (red). Notably, West Nile Virus is vectored by both Aedes and Culex species. Predicted vectors of Zika are shown by bolded links in black. The inset shows predicted vectors of Zika and species names, ordered by the model’s propensity scores. Included flaviviruses are Banzi virus (BANV), Bouboui virus (BOUV), dengue virus strains 1, 2, 3 and 4 (DENV-1,2,3,4), Edge Hill virus (EHV), Ilheus virus (ILHV), Israel turkey meningoencephalomyelitis virus (ITV), Japanese encephalitis virus (JEV), Kedougou virus (KEDV), Kokobera virus (KOKV), Kunjin virus (KUNV), Murray Valley encephalitis virus (MVEV), Rocio virus (ROCV), St. Louis encephalitis virus (SLEV), Spondwendi virus (SPOV), Stratford virus (STRV), Uganda S virus (UGSV), Wesselsbron virus (WESSV), West Nile Virus (WNV), yellow fever virus (YFV), and Zika virus (ZIKV).

https://doi.org/10.7554/eLife.22053.003

Our ensemble of BRT models trained on common vector and virus traits predicted mosquito vector-virus pairs in the test dataset with high accuracy (AUC = 0.92 ± 0.02; sensitivity = 0.858 ± 0.04; specificity = 0.872 ± 0.04). Due to non-monotonicity and existence of interactions among predictor variables within our model, one cannot make general statements about the directionality of effect. Thus, we focus on the relative importance of different variables to model performance. The most important variable for accurately predicting the presence of vector-virus pair was the subgenus of the mosquito species, followed by continental range (e.g. continents on which species are present). The number of viruses vectored by a mosquito species and number of mosquito vectors of a virus were the third and fifth most important variables, respectively. Unsurprisingly, this suggests that, when controlling for other variables, mosquitoes and viruses with more known vector-virus pairs (i.e., more viruses vectored and more hosts infected, respectively), are more likely to be part of a predicted pair by the model. Mosquito ecological traits such as larval habitat and salinity tolerance were generally less important than a species’ phylogeny or geographic range (Figure 2).

Variable importance by permutation, averaged over 25 models.

Because some categorical variables were treated as binary by our model (i.e. continental range), the relative importance of each binary variable was summed to result in the overall importance of the categorical variable. Mosquito and virus traits are shown in blue and maroon, respectively. Error bars represent the standard error from 25 models.

https://doi.org/10.7554/eLife.22053.004

When applied to the 180 potential mosquito-Zika pairs, the model predicted thirty-five vectors to be ranked above the threshold (set at the value of the lowest-ranked known vector), for a total of nine known vectors and twenty-six novel, predicted mosquito vectors of Zika (Table 1). Of these vectors, there were twenty-four Aedes species, nine Culex species, one Psorophora species, and one Runchomyia species. The GBM model’s top two ranked vectors for Zika are the most highly-suspected vectors of Zika virus, Ae. aegypti and Ae. albopictus.

Model validation

Our supplementary and primary models generally concur and their ranking of potential Zika virus vectors are highly correlated (ρ = 0.508 and ρ = 0.693 on raw and thresholded predictions, respectively). As one might expect, the supplementary model assigned fewer scores of low propensity (Appendix 1—figure 2), suggesting that incorporating this additional uncertainty in the training dataset eroded the model's ability to distinguish negative links. The supplementary model’s performance on the testing data (AUC = 0.84 ± 0.02), however, indicates that the additional uncertainty did not impede model performance.

When trained on ‘leave-one-out’ datasets, all three models were able to predict the testing data with high accuracy (AUC = 0.91, AUC = 0.91, AUC = 0.92 for West Nile, dengue, and yellow fever viruses, respectively). Performance varied when models were validated against predictions of ‘known outcomes’. A model trained without West Nile virus predicted highly linked vectors reasonably well (AUC = 0.69), however it assigned low scores to rarer ‘known’ vectors, such as Culiseta inornata, which was only associated with West Nile virus. Similarly, the model trained on the dengue-omitted dataset predicted training data and vectors of dengue itself with high accuracy (AUC = 0.92). While the model trained without yellow fever performed well on the testing data, it performed poorly when predicting vectors of yellow fever virus (AUC = 0.47). Unlike West Nile and dengue viruses, the majority of the known vectors of yellow fever are only associated with yellow fever (i.e. a single vector-virus link), and so were excluded completely from the training data when all yellow fever links were omitted. Additionally, several of the vector species are of the Haemagogus genus, which was completely absent from the training data. Given the importance of phylogeny of the vector species in predicting vector-virus links, it follows that a dataset with a novel subgenus would be difficult for the model to predict on, resulting in low model performance. The low performance of this model illustrates that incorporating common traits and additional vector-virus links improves model prediction. When traits were not available in the training dataset, model performance was much lower, suggesting that there exists a statistical association between a vectors’ traits and its ability to transmit a virus.

Discussion

Zika virus is unprecedented among emerging arboviruses in its combination of severe public health hazard, rapid spread, and poor scientific understanding. Particularly crucial to public health preparedness is knowledge about the geographic extent of potentially at risk populations and local environmental conditions for transmission, which are determined by the presence of competent vectors. Until now, identifying additional competent vector species has been a low priority because Zika virus has historically been geographically restricted to a narrow region of equatorial Africa and Asia (Petersen et al., 2016), and the mild symptoms of infection made its range expansion since the 1950’s relatively unremarkable. However, with its relatively recent and rapid expansion into the Americas and its association with severe neurological disorders, the prediction of potential disease vectors in non-endemic areas has become a matter of critical public health importance. We identify these potential vector species by developing a data-driven model that identifies candidate vector species of Zika virus by leveraging data on traits of mosquito vectors and their flaviviruses. We suggest that empirical work should prioritize these species in their evaluation of vector competence of mosquitoes for Zika virus.

Our model predicts that fewer than one third of the potential mosquito vectors of Zika virus have been identified, with over twenty-five additional mosquito species worldwide that may have the capacity to contribute to transmission. The continuing focus in the published literature on two species known to transmit Zika virus (Ae. aegypti and Ae. albopictus) ignores the potential role of other vectors, potentially misrepresenting the spatial extent of risk. In particular, four species predicted by our model to be competent vectors – Ae. vexans, Culex quinquefasciatus, Cx. pipiens, and Cx. tarsalis – are found throughout the continental United States. Further, the three Culex species are primary vectors of West Nile virus (Farajollahi et al., 2011). Cx. quinquefasciatus and Cx. pipiens were ranked 3rd and 17th by our model, respectively, and together these species were the highest-ranking species endemic to the United States after the known vectors (Ae. aegypti and Ae. albopictus). Cx. quinquefasciatus has previously been implicated as an important vector of encephalitic flaviviruses, specifically West Nile virus and St. Louis encephalitis (Turell et al., 2005; Hayes et al., 2005), and a hybridization of the species with Cx. pipiens readily bites humans (Fonseca et al., 2004). The empirical data available on the vector competence of Cx. pipiens and Cx. quinquefasciatus is currently mixed, with some studies finding evidence for virus transmission and others not (Guo et al., 2016; Aliota et al., 2016; Fernandes et al., 2016; Huang et al., 2016). These results suggest, in combination with evidence for significant genotype x genotype effects on the vector competence of Ae. aegypti and Ae. albopictus to transmit Zika (Chouin-Carneiro et al., 2016), that the vector competence of Cx. pipiens and Cx. quinquefasciatus for Zika virus could be highly dependent upon the genetic background of the mosquito-virus pairing, as well as local environmental conditions. Thus, considering their anthropophilic natures and wide geographic ranges, Cx. quinquefasciatus and Cx. pipiens could potentially play a larger role in the transmission of Zika in the continental United States. Further experimental research into the competence of populations of Cx. pipiens to transmit Zika virus across a wider geographic range is therefore highly recommended, and should be prioritized.

The vectors predicted by our model have a combined geographic range much larger than that of the currently suspected vectors of Zika (Figure 3), suggesting that, were these species to be confirmed as vectors, a larger population may be at risk of Zika infection than depicted by maps focusing solely on Ae. aegypti and Ae. albopictus. The range of Cx. pipiens includes the Pacific Northwest and the upper mid-West, areas that are not within the known range of Ae. aegypti or Ae. albopictus (Darsie and Ward, 2005). Furthermore, Ae. vexans, another predicted vector of Zika virus, is found throughout the continental US and the range of Cx. tarsalis extends along the entire West coast (Darsie and Ward, 2005). On a finer scale, these species use a more diverse set of habitats, with Ae. aegypti and Cx. quinquefasciatus mainly breeding in artificial containers, and Ae. vexans and Ae. albopictus being relatively indiscriminate in their breeding sites, including breeding in natural sites such as tree holes and swamps. Therefore, in addition to the wider geographic region supporting potential vectors, these findings suggest that both rural and urban areas could serve as habitat for potential vectors of Zika. We recommend experimental tests of these species for competency to transmit Zika virus, because a confirmation of these vectors would necessitate expanding public health efforts to these areas not currently considered at risk.

Distribution maps of predicted vectors of Zika virus in the continental US.

Maps of Aedes species are based on Centers for disease control and prevention (2016). All other species’ distributions are georectified maps from Darsie and Ward (2005).

https://doi.org/10.7554/eLife.22053.005

While transmission requires a competent vector, vector competence does not necessarily equal transmission risk or inform vectorial capacity. There are many biological factors that, in conjunction with positive vector competence, determine a vector’s role in disease transmission. For example, although Ae. aegypti mosquitoes are efficient vectors of West Nile virus, they prefer to feed on humans, which are dead-head hosts for the disease, and therefore have low potential to serve as a vector (Turell et al., 2005). Psorophora ferox, although predicted by our model as a potential vector of Zika virus, would likely play a limited role in transmission because it rarely feeds on humans (Molaei et al., 2008). Additionally, vector competence is dynamic, and may be mediated by environmental factors that influence viral development and mosquito immunity (Muturi and Alto, 2011). Therefore, our list of potential vectors of Zika represents a comprehensive starting point, which should be furthered narrowed by empirical work and consideration of biological details that impact transmission dynamics. Given the severe neurological side-effects of Zika virus infection, beginning with the most conservative method of vector prediction ensures that risk is not underestimated, and allows public health agencies to interpret the possibility of Zika transmission given local conditions.

Our model serves as a starting point to streamlining empirical efforts to identify areas and populations at risk for Zika transmission. While our model enables data-driven predictions about the geographic area at potential risk of Zika transmission, subsequent empirical work investigating Zika vector competence and transmission efficiency is required for model validation, and to inform future analyses of transmission dynamics. For example, in spite of its low transmission efficiency in certain geographic regions (Chouin-Carneiro et al., 2016), Ae. aegypti is anthropophilic (Powell and Tabachnick, 2013), and may therefore pose a greater risk of human-to-human Zika virus transmission than mosquitoes that bite a wider variety of animals. On the other hand, mosquito species that prefer certain hosts in rural environments are known to alter their feeding behaviors to bite alternative hosts (e.g., humans and rodents) in urban settings, due to changes in host community composition (Chaves et al., 2010). Environmental factors such as precipitation and temperature directly influence mosquito populations, and determine the density of vectors in a given area (Thomson et al., 2006), an important factor in transmission risk. Additionally, socio-economic factors such as housing type and lifestyle can decrease a populations’ contact with mosquito vectors, and lower the risk of transmission to humans (Moreno-Madriñán and Turell, 2017). Effective risk modeling and forecasting the range expansion of Zika virus in the United States will depend on validating the vector status of these species, as well as resolving behavioral and biological details that impact transmission dynamics.

Although we developed this model with Zika virus in mind, our findings have implications for other emerging flaviviruses and contribute to the recently developed methodology applying machine learning methods to the prediction of unknown agents of infectious diseases. This technique has been used to predict rodent reservoirs of disease (Han et al., 2015) and bat carriers of filoviruses (Han et al., 2016) by training models with host-specific data. Our model, however, incorporates additional data by constructing a vector-virus network that is used to inform predictions of vector-virus associations. The combination of common virus traits with vector-specific traits enabled us to predict potential mosquito vectors of specific flaviviruses, and to train the model on additional information distributed throughout the flavivirus-mosquito network.

Uncertainty in our model arises through uncertainty inherent in our datasets. Vector status is not static (e.g. mutation in the chikungunya virus to increase transmission by Ae. albopictus [Weaver and Forrester, 2015]) and can vary across vector populations (Bennett et al., 2002). When incorporating uncertainty in vector status through our supplementary model, our predictions generally agreed with that of our original model. However, the increased uncertainty did reduce the models’ ability to distinguish negative links, resulting in higher uncertainty in propensity scores (as measured by standard deviation) and a larger number of predicted vectors. Additionally, the model performs poorly when predicting on vector-virus links with trait levels not included in the training data set, as was the case when omitting yellow fever virus. Another source of uncertainty is regarding vector and virus traits. In addition to intraspecific variation in biological traits, many vectors are understudied, and common traits such as biting activity are unknown to the level of species. Additional study into the behavior and biology of less common vector species would increase the accuracy of prediction techniques such as this, and allow for a better of understanding of species’ potential role as vectors.

Interestingly, our constructed flavivirus-mosquito network generally concurs with the proposed dichotomy of Aedes species vectoring hemorrhagic or febrile arboviruses and Culex species vectoring neurological or encephalitic viruses (Grard et al., 2010) (Figure 1). However, there are several exceptions to this trend, notably West Nile virus, which is vectored by several Aedes species. Additionally, our model predicts several Culex species to be possible vectors of Zika virus. While this may initially seem contrary to the common phylogenetic pairing of vectors and viruses noted above, Zika’s symptoms, like West Nile virus, are both febrile and neurological. Thus, its symptoms do not follow the conventional hemorrhagic/encephalitic division. The ability of Zika virus to be vectored by a diversity of mosquito vectors could have important public health consequences, as it may expand both the geographic range and seasonal transmission risk of Zika virus, and warrants further empirical investigation.

Considering our predictions of potential vector species and their combined ranges, species on the candidate vector list need to be validated to inform the response to Zika virus. Vector control efforts that target Aedes species exclusively may ultimately be unsuccessful in controlling transmission of Zika because they do not control other, unknown vectors. For example, the release of genetically modified Ae. aegypti to control vector density through sterile insect technique is species-specific and would not control alternative vectors (Alphey et al., 2010). Additionally, species’ habitat preferences differ, and control efforts based singularly on reducing Aedes larval habitat will not be as successful at controlling Cx. quinquefasciatus populations (Rey et al., 2006). Predicted vectors of Zika virus must be empirically tested and, if confirmed, vector control efforts would need to respond by widening their focus to control the abundance of all predicted vectors of Zika virus. Similarly, if control efforts are to include all areas at potential risk of disease transmission, public health efforts would need to expand to address regions such as the northern Midwest that fall within the range of the additional vector species predicted by our model. An understanding of the capacity of mosquito species to vector Zika virus is necessary to prepare for the potential establishment of Zika virus in the United States, and we recommend that experimental work start with this list of candidate vector species.

Materials and methods

Data collection and feature construction

Our dataset comprised a matrix of vector-virus pairs relating all known flaviviruses and their mosquito vectors. To construct this matrix, we first compiled a list of mosquito-borne flaviviruses to include in our study (Van Regenmortel et al., 2000; Kuno et al., 1998; Cook and Holmes, 2006). Viruses that only infect mosquitoes and are not known to infect humans were not included. Using this list, we constructed a mosquito-virus pair matrix based on the Global Infectious Diseases and Epidemiology Network database (GIDEON, 2016), the International Catalog of Arboviruses Including Certain Other Viruses of Vertebrates (ArboCat) (Karabatsos, 1985), The Encyclopedia of Medical and Veterinary Entomology (Russell et al., 2013)and Mackenzie et al. (2012).

We defined a known vector-virus pair as one for which the full transmission cycle (i.e, infection of mosquito via an infected host (mammal or avian) or bloodmeal that is able to be transmitted via saliva) has been observed. Basing vector competence on isolation or intrathoracic injection bypasses several important barriers to transmission (Hardy et al., 1983), and may not be true evidence of a mosquito’s ability to transmit an arbovirus. We found our definition to be more conservative than that which is commonly used in disease databases (e.g. Global Infectious Diseases and Epidemiology Network database), which often assumes isolation from wild-caught mosquitoes to be evidence of a mosquito’s role as a vector. Therefore, a supplementary analysis investigates the robustness of our findings with regards to uncertainty in vector status by comparing the analysis reported in the main text to a second analysis in which any kind of evidence for association, including merely isolating the virus in wild-caught mosquitoes, is taken as a basis for connection in the virus-vector network (see Appendix 1 for analysis and results).

Fifteen mosquito traits (Appendix 2—table 1) and twelve virus traits (Appendix 2—table 2) were collected from the literature. For the mosquito species, the geographic range was defined as the number of countries in which the species has been collected, based on Walter Reed Biosystematics Unit, (2016). While there are uncertainties in species’ ranges due to false absences, this represents the most comprehensive, standardized dataset available that includes both rare and common mosquito species. A species’ continental extent was recorded as a binary value of its presence by continent. A species’ host range was defined as the number of taxonomic classes the species is known to feed on, with the Mammalia class further split into non-human primates and other mammals, because of the important role primates play in zoonotic spillovers of vector-borne disease (e.g. dengue, chikungunya, yellow fever, and Zika viruses) (Weaver, 2005; Diallo et al., 2005; Weaver et al., 2016). The total number of unique flaviviruses observed per mosquito species was calculated from our mosquito-flavivirus matrix. All other traits were based on consensus in the literature (see Appendix III for sources by species). For three traits – urban preference, endophily (a proclivity to bite indoors), and salinity tolerance – if evidence of that trait for a mosquito was not found in the literature, it was assumed to be negative.

Table 1

Predicted vectors of Zika virus, as reported by our model. Mosquito species endemic to the continental United States are bolded. A species is defined as a known vector of Zika virus if a full transmission cycle (see main text) has been observed.

https://doi.org/10.7554/eLife.22053.006
SpeciesGBM prediction ±SDKnown vector?
Aedes aegypti0.81 ± 0.12Yes
Ae. albopictus0.54 ± 0.14Yes
Culex quinquefasciatus0.38 ± 0.14No
Ae. polynesiensis0.36 ± 0.13No
Ae. scutellaris0.33 ± 0.13No
Ae. africanus0.32 ± 0.11No
Ae. furcifer0.31 ± 0.16Yes
Ae. vittatus0.30 ± 0.20Yes
Ae. taylori0.30 ± 0.16Yes
Ae. luteocephalus0.25 ± 0.12Yes
Ae. tarsalis0.18 ± 0.11Yes
Ae. metallicus0.16 ± 0.08No
Ae. minutus0.16 ±0.09No
Ae. opok0.14 ± 0.06No
Ae. bromeliae0.11 ± 0.06No
Ae. scapularis0.10 ± 0.04No
Cx. pipiens0.10 ± 0.04No
Ae. hensilli0.10 ± 0.06Yes
Ae. vigilax0.10 ± 0.05No
Cx. annulirostrix0.08 ± 0.03No
Psorophora ferox0.08 ± 0.05No
Cx. rubinotus0.08 ± 0.07No
Cx. tarsalis0.08 ± 0.03No
Ae. occidentalis0.08 ± 0.05No
Ae. flavicolis0.07 ± 0.04No
Ae. serratus0.07 ± 0.04No
Cx. p. molestus0.07 ± 0.04No
Ae. vexans0.06 ± 0.04No
Cx. neavei0.06 ± 0.02No
Runchomyia frontosa0.06 ± 0.04No
Ae. neoafricanus0.06 ± 0.03No
Ae. chemulpoensis0.06 ± 0.03No
Cx. vishnui0.05 ± 0.01No
Cx. tritaeniorhynchus0.05 ± 0.01No
Ae. fowleri0.04 ± 0.03Yes

We collected data on the following virus traits: host range (Mahy, 2009; Mackenzie et al., 2012; Chambers and Monath, 2003; Cook and Zumla, 2009b), disease severity (Mackenzie et al., 2012), human illness (Chambers and Monath, 2003; Cook and Zumla, 2009), the presence of a mutated envelope protein, which controls viral entry into cells (Grard et al., 2010), year of isolation (Karabatsos, 1985), and host range (Karabatsos, 1985). Disease severity was based on Mackenzie et al. (2012), ranging from no known symptoms (e.g. Kunjin virus) to severe symptoms and significant human mortality (e.g. yellow fever virus). For each virus, vector range was calculated as the number of mosquito species for which the full transmission cycle has been observed. Genome length was calculated as the mean of all complete genome sequences listed for each flavivirus in the Virus Pathogen Database and Analysis Resource (http://www.viprbrc.org/). For more recently discovered flaviviruses not yet cataloged in the above databases (i.e., New Mapoon Virus, Iquape virus), viral traits were gathered from the primary literature (sources listed in Appendix 3).

Predictive model

Following Han et al. (2015), boosted regression trees (BRT) (Friedman, 2001) were used to fit a logistic-like predictive model relating the status of all possible virus-vector pairs (0: not associated, 1: associated) to a predictor matrix comprising the traits of the mosquito and virus traits in each pair. Boosted regression trees circumvent many issues associated with traditional regression analysis (Elith et al., 2008), allowing for complex variable interactions, collinearity, non-linear relationships between covariates and response variables, and missing data. Additionally, this technique performs well in comparison with other logistic regression approaches (Friedman, 2001). Trained boosted regression tree models are dependent on the split between training and testing data, such that each model might predict slightly different propensity values. To address this, we trained an ensemble of 25 internally cross-validated BRT models on independent partitions of training and testing data. The resulting model demonstrated low variance in relative variable importance and overall model accuracy, suggesting models all converged to a similar result.

Prior to the analysis of each model, we randomly split the data into training (70%) and test (30%) sets while preserving the proportion of positive labels (known associations) in each of the training and test sets. Models were trained using the gbm package in R (Ridgeway, 2015), with the maximum number of trees set to 25,000, a learning rate of 0.001, and an interaction depth of 5. To correct for optimistic bias (Smith et al., 2014), we performed 10-fold cross validation and chose a bag fraction of 50% of the training data for each iteration of the model. We estimated the performance of each individual model with three metrics: Area Under the Receiver Operator Curve, specificity, and sensitivity. For specificity and sensitivity, which require a preset threshold, we thresholded predictions on the testing data based on the value which maximized the sum of the sensitivity and specificity, a threshold robust to the ratio of presence to background points in presence-only datasets (Liu et al., 2016). Variable importance was quantified by permutation (Breiman, 2001) to assess the relative contribution of virus and vector traits to the propensity for a virus and vector to form a pair. Because we transformed many categorical variables into binary variables (e.g., continental range as binary presence or absence by continent), the sum of the relative importance for each binary feature was summed to obtain a single value for the entire variable.

Each of our twenty-five trained models was then used to predict novel mosquito vectors of Zika by applying the trained model to a data set consisting of the virus traits of Zika paired with the traits of all mosquitoes for which flaviviruses have been isolated from wild caught individuals, and, depending on the species, may or may not have been tested in full transmission cycle experiments (a total of 180 mosquito species). This expanded dataset allowed us to predict over a large number of mosquito species, while reasonably limiting our dataset to those species suspected of transmitting flaviviruses. The output of this model was a propensity score ranging from 0 to 1. In our case, the final propensity score for each vector was the mean propensity score assigned by the twenty-five models. To label unobserved edges, we thresholded propensity scores at the value of lowest ranked known vector (Liu et al., 2013).

Model validation

In addition to conventional performance metrics, we conducted additional analyses to further validate both this method of prediction, and our model specifically. To account for uncertainty in the vector-virus links in our initial matrix, we repeated our analysis for a vector-virus matrix with a less conservative definition of a positive link (field isolation and above), referred to as our supplementary model. Vector competence is a dynamic trait, and there exists significant intraspecific variation in the ability of a vector to transmit a virus for certain species of mosquitoes (Diallo et al., 2005; Gubler et al., 1979). Our supplementary model is based on a less conservative definition of vector competence and includes species implicated as vectors, but not yet verified through laboratory competence studies, and therefore accounts for additional uncertainty such as intraspecific variation.

While this approach is well-tested in epidemiological applications (Parascandola, 2004), it has only recently been applied to predict ecological associations, and, as such, has limitations unique to this application. To further evaluate this prediction method, we performed a modified ‘leave-one-out’ analysis, whereby we trained a model to a dataset from which a well-studied virus had been omitted, and then predicted vectors for this virus and compared them against a list of known vectors. We repeated this analysis for West Nile, dengue, and yellow fever viruses, following the same method of training as for our original model. While this analysis differs from our original method, it provides a more stringent evaluation of this method of prediction because the model is trained on an incomplete dataset and predicts on unfamiliar data, a more difficult task than that posed to our original model.

Appendix 1

Comparison model trained on virus isolation data

The primary model is trained on vector-virus pairs for which the full transmission cycle has been observed. However, many sources, such as the Global Infectious Diseases and Epidemiology Network database (GIDEON), interpret isolation of a virus in wild-caught mosquitoes as evidence of a mosquito’s role as a vector. In order to investigate the robustness of our findings, we conducted a supplementary analysis in which any evidence for association, including isolation of the virus, is used as the basis for a link in the vector-virus network.

Data collection

As in the primary model, the mosquito-virus pair matrix was constructed based on the Global Infectious Diseases and Epidemiology Network database (GIDEON, 2016), the International Catalog of Arboviruses Including Certain Other Viruses of Vertebrates (ArboCat) (Karabatsos, 1985), The Encyclopedia of Medical and Veterinary Entomology (Russell et al., 2013) and (Mackenzie et al., 2012). This resulted in a dataset containing 180 mosquito species and 37 viruses, for a total of 334 vector-virus pairs. The vector and virus trait datasets were identical to those used in the primary model (see Appendix 2 for lists of traits).

Predictive model

We used boosted regression trees (Friedman, 2001) to fit a logistic-like predictive model relating the status of all possible virus-vector pairs (0: not associated, 1:associated) to a predictor matrix comprising the traits of the mosquito and virus traits in each pair. We fit a total of 25 models, applying different training and testing datasets to each, to reduce the dependence dependent on the split between training and testing data. Prior to the analysis of each model, we randomly split the data into training (70%) and test (30%) sets while preserving the proportion of positive labels in each of the training and test sets. Models were trained using the gbm package in R (Ridgeway, 2015), with the maximum number of trees set to 25,000 and a learning rate of 0.001. To correct for optimistic bias (Smith et al., 2014), we performed 10-fold cross validation and bagged 50% of the training data for each iteration of the model. These methods are identical to those used to train the primary model. We quantified variable importance by permutation (Breiman, 2001) to assess the relative contribution of virus and vector traits to the propensity for a virus and vector to form a pair.Each of our twenty-five trained models was then used to predict novel mosquito vectors of Zika over the whole virus-vector pair dataset, resulting in twenty-five propensity values assigned to each mosquito species, of which we took the mean. Our prediction dataset, therefore, consisted of the common virus traits of Zika paired with the common traits of all mosquitoes in our flavivirus dataset, for a total of 180 species. The output of this model was a propensity score ranging from 0 to 1. In our case, the final propensity score for each vector was the mean propensity score assigned by the twenty-five models. To label unobserved edges, we thresholded propensity at the value of lowest ranked known vector (Liu et al., 2013).

Results

Boosted regression models trained on the weakest evidence of association accurately predicted mosquito vector-virus associations in the test dataset (AUC= 0.84 ±0.02). When thresholded at the value of the lowest ranked known vector, the model predicted 66 potential vectors of ZIKV, including 42 unknown vectors LABEL:table:predictions. The majority of predicted vectors were Aedes species (39 species), with Culex as the second most predicted genus (15 species). It included all but three of the vectors predicted by the main model (Ae. occidentalis, Ru. frontosa, Cx. rubinotus).

Model comparisons

Our supplementary and primary models, trained on virus isolation and above and full transmission cycle, respectively, generally concur. The models are fairly correlated (Spearman’s coefficient, ρ=0.508 when considering the propensities of all 180 species 1. However, when only comparing the correlation of propensities between those vectors above the threshold of lowest ranked known vector, the models become much more correlated (ρ=0.693). This suggests that our model has a higher sensitivity than specificity, and is better able to predict those vectors that are competent for ZIKV than those that are not. The predictive accuracy of our supplementary model was slightly lower than our primary model. However, this may be an indirect effect of a lower positive-negative label ratio in the dataset used in the primary model, which can artificially inflate AUC values (Lobo et al., 2008).

The models differ in their ability to differentiate between vectors and non-vectors. The distribution of propensities for our main model is more skewed towards lower propensity values than is the supplementary model 2. This is logical, as the dataset used to train the main model contains a higher proportion of zeros (e.g. vector-virus pairs with no known association) than the supplementary model. The difference in distributions is accounted for by a similar discrepancy in threshold propensity values based on the lowest ranked known vector. The main model, which has a higher frequency of near-zero propensities, uses a lower threshold value than the supplementary model, however both thresholds qualitatively lie above the majority of the distributions.

Conclusion

In summary, our supplementary model predicts which mosquito species may test positive for ZIKV through isolation in wild-caught individuals. As isolation can be understood as evidence of a vector’s role in transmission of a disease, our supplementary model may also be interpreted as a ranking of potential vectors of ZIKV, similar to our main model. In fact, both models are well correlated in their ranking of species, although the main model, which trains on fewer vector-virus links, predicts fewer vectors than the supplementary model. Those species predicted by both models, such as Cx. quinquefasciatus and Ae. vexans, should be prioritized for further research on their competency to transmit ZIKV. Furthermore, as suggested by the main model, the current geographic range at risk for ZIKV transmission in the United States should be expanded to include the range of these species ranked highly by both our main and supplementary models.

Appendix 1—table 1

Vector predictions by the supplementary model.

https://doi.org/10.7554/eLife.22053.007
Vector
GBM PredictionSD
Aedes aegypti0.840.06
Aedes albopictus0.810.07
Aedes vittatus0.760.10
Aedes africanus0.700.11
Aedes taylori0.650.14
Aedes furcifer0.650.14
Aedes luteocephalus0.590.12
Aedes metallicus0.590.13
Aedes opok0.580.13
Culex quinquefasciatus0.560.13
Aedes tarsalis0.560.12
Aedes scutellaris0.560.11
Aedes minutus0.550.12
Aedes polynesiensis0.530.11
Mansonia uniformis0.520.12
Aedes fowleri0.480.14
Aedes vexans0.460.11
Aedes dalzieli0.450.13
Culex annulirostris0.450.08
Mansonia africana0.420.12
Psorophora ferox0.390.14
Culex tarsalis0.380.09
Culex tritaeniorhynchus0.370.08
Culex pipiens0.370.13
Culex neavei0.340.06
Aedes vigilax0.340.07
Aedes flavicollis0.330.14
Aedes scapularis0.310.07
Aedes taeniarostris0.310.13
Aedes jamoti0.310.13
Aedes circumluteolus0.300.13
Eretmapodites inornatus0.300.15
Aedes cumminsii0.290.11
Culex vishnui0.280.05
Aedes lineatopennis0.280.11
Aedes neoafricanus0.270.11
Aedes bromeliae0.260.10
Culex guiarti0.260.06
Culex perfuscus0.260.06
Aedes stokesi0.260.12
Culex telesilla0.250.06
Anopheles gambiae0.240.11
Sabethes chloropterus0.240.11
Aedes hensilli0.240.09
Aedes serratus0.230.06
Aedes chemulpoensis0.230.08
Aedes normanensis0.230.06
Culex bitaeniorhynchus0.220.09
Culex pseudovishnui0.220.05
Aedes argenteopunctatus0.210.06
Wyeomyia vanduzeei0.210.15
Culex p. molestus0.210.06
Culex salinarius0.200.04
Aedes grahami0.190.15
Anopheles coustani0.190.08
Aedes longipalpis0.180.18
Uranotaenia sapphirina0.170.08
Aedes domesticus0.170.06
Aedes abnormalis0.170.06
Aedes natronius0.170.06
Eretmapodites chrysogaster0.170.08
Aedes mcintoshi0.170.06
Aedes ochraceus0.160.06
Culex fatigans0.160.07
Anopheles amictus0.160.06
Eretmapodites quinquevittatus0.160.08
Appendix 1—figure 1
Propensity values of the main and supplementary models.

Dashed lines represent corresponding threshold values for each model based on lowest ranked known vector propensities.

https://doi.org/10.7554/eLife.22053.008
Appendix 1—figure 2
Distribution of propensity values for the main and supplementary models.

Dashed lines represent corresponding threshold values for each model based on lowest ranked known vector propensities.

https://doi.org/10.7554/eLife.22053.009

Appendix 2

Tables of vector and virus traits

Appendix 2—table 1

Table of mosquito traits used in model.

https://doi.org/10.7554/eLife.22053.010
TraitTypeSubcategories
AnthropophilybinaryNA
SubgenusfactorNA
Host breadthnumericNA
Host rangebinary (x4)Primate, Non-Primate Mammal, Bird, Cold-Blooded Vertebrate
Geographic areanumericNA
Continental rangebinary (x8)Africa, Middle East, Australia, Pacific, Asia, Europe, North America, South America
Biting timebinary (x4)Dawn, Day, Dusk, Night
Artificia container breederbinaryNA
Oviposition sitebinary (x8)Treehole, Natural Container, Permanent Fresh Water, Rockhole, Marsh, Swamp, Temporary Ground Pools, Rice Paddy
Habitat discriminationnumericNA
Salinity tolerancebinaryNA
Habitat permanencebinaryNA
Urban preferencebinaryNA
EndophilybinaryNA
No. of flaviviruses vectorednumericNA
Appendix 2—table 2

Table of virus traits used in model.

https://doi.org/10.7554/eLife.22053.011
TraitTypeSubcategories
GroupfactorJapanese Encephalitis, Ntaya, Yellow Fever, Aroa, Dengue, Kokobera, Spondweni
Continental rangebinary(x8)Africa, Middle East, Australia, Pacific, Asia, Europe, North America, South America
CladefactorVI, VII, IX, X, XI, XII, XIV,
Year isolatednumericNA
Mutated envelopebinaryNA
Host breadthnumericNA
Host Rangebinary(x6)Human, Non-Human Primate, Rodent, Other Mammal, Bird, Marsupial
Mosquito vector breadthnumericNA
Vectored by other arthropodsbinaryNA
Disease symptomsbinary (x2)Encephalitis, Fever
Disease severitynumericNA
Genome lengthnumericNA

Appendix 3

Primary sources used for vector and virus traits

Appendix 3—table 1

Primary sources for mosquito traits.

https://doi.org/10.7554/eLife.22053.012
Mosquito speciesSources
Aedeomyia africanaRobert et al. (1998), Harbach (2015), Omondi et al. (2015)
Aedeomyia catastictaHarbach (2015), Jansen et al. (2009), Wright et al. (1981)
Aedes abnormalisIwuala (1981)
Aedes aegyptiHalstead (2008), Ramasamy et al. (2011)
Aedes africanusHaddow (1961)
Aedes albopictusRamasamy et al. (2011)
Aedes alternansNSW Health (2016), Russell et al. (2013), Knight et al. (2012)
Aedes argenteopunctatusHarbach (2015), Fontenille et al. (1998)
Aedes bancroftianusNSW Health (2016), Russell (1986), Harbach (2015)
Aedes bromeliaeBennett et al. (2015), Beran (1994), Digoutte (1999)
Aedes caballusHarbach (2015), Steyn and Schulz (1955)
Aedes canadensisCarpenter and LaCasse (1974), Andreadis et al. (2004)
Aedes cantansRenshaw et al. (1994, 1995), Service (1993)
Aedes cantatorGiberson et al. (2007)
Aedes chemulpoensisFeng (1983)
Aedes cinereusMorrison and Andreadis (1992), Anderson et al. (2007), Becker and Neumann (1983), Molaei et al. (2008)
Aedes circumluteolusJupp and McIntosh (1987), Paterson et al. (1964), Chandler et al. (1975)
Aedes cumminsiLane and Crosskey (2012)
Aedes curtipesHarbach (2015), MacDonald et al. (1965), Knight and Hull (1953)
Aedes dalzieliFontenille et al. (1998)
Aedes domesticusHarbach (2015), Lane and Crosskey (2012), Geoffroy (1987)
Aedes dorsalisAldemir et al. (2010), Wang et al. (2012)
Aedes flavicolisReinert (1970)
Aedes fluviatilisMultini et al. (2015), Baton et al. (2013), Reinert et al. (2008)
Aedes fowleri(Boussès et al., 2013)
Aedes furciferBeran (1994), Hopkins (1952)
Aedes grahamiHarbach (2015)
Aedes hensilliLedermann et al. (2014), Bohart and Ingram (1946)
Aedes ingramiLane and Crosskey (2012), Haddow (1946b, 1964, 1942)
Aedes jamotiHarbach (2015), Le Berre and Hamon (1961)
Aedes japonicusKaufman and Fonseca (2014), Kampen and Werner (2014)
Aedes juppiHarbach (2015), Jupp and Kemp (1998)
Aedes koreicusHarbach (2015), Montarsi et al. (2013), Medlock et al. (2015)
Aedes lineatopennisHarbach (2015), Amerasinghe and Indrajith (1995), Jupp (1967), Linthicum et al. (1985)
Aedes longipalpisHarbach (2015)
Aedes luteocephalusDiallo et al. (2012a), Service (1965b), Boorman (1961)
Aedes mcintoshiWalter Reed Biosystematics Unit (2016), Harbach (2015)
Aedes mediolineatusHarbach (2015)
Aedes melanimonWalter Reed Biosystematics Unit (2016), Barker et al. (2009), Chapman (1960)
Aedes metallicusHarbach (2015), Beran (1994)
Aedes minutusHarbach (2015), Diallo et al. (2012b)
Aedes natroniusHarbach (2015)
Aedes neoafricanusHarbach (2015), Diallo et al. (2012b), Hervy et al. (1986)
Aedes normanensisNSW Health (2016), Hearnden and Kay (1995)
Aedes notoscriptusNSW Health (2016), Jansen et al. (2015), Nicholson et al. (2015), Derraik et al. (2007), Frances et al. (2002)
Aedes occidentalisHarbach (2015), Evans (1926)
Aedes ochraceusCorbet (1962), Lutomiah et al. (2014)
Aedes opokBeran (1994), Herve et al. (1975), Germain et al. (1976)
Aedes polynesiensisYoung (2007)
Aedes procaxNSW Health (2016), Ryan and Kay (2000)
Aedes scapularisForattini et al. (1988)
Aedes scutellarisPenn (1947)
Aedes serratusGuimarães et al. (2000), Cardoso et al. (2010)
Aedes simulansHarbach (2015)
Aedes sollicitansGiberson et al. (2007), Carpenter and LaCasse (1974), Crans and Sprenger (1996), Crans et al. (1996)
Aedes stokesiHarbach (2015), Reinert (1986)
Aedes taeniarostrisEastwood et al. (2013)
Aedes tarsalisEllis et al. (2007)
Aedes tayloriWalter Reed Biosystematics Unit (2016)
Aedes togoiTsunoda et al. (2012), Lee and Hong (1995)
Aedes tremulusKay et al. (2000), Webb et al. (2016)
Aedes trivittatusCarpenter and LaCasse (1974), Andreadis et al. (2004)
Aedes vexansBoxmeyer and Palchick (1999), Aldemir et al. (2010)
Aedes vigilaxNSW Health (2016), Chapman et al. (1999)
Aedes vittatusBoorman (1961), Selvaraj and Dwarakanath (1992)
Anopheles amictusHearnden and Kay (1995)
Anopheles barbirostrisSriwichai et al. (2016), Amerasinghe and Indrajith (1995), Bashar et al. (2012)
Anopheles coustaniFornadel et al. (2011), Mwangangi et al. (2013), Muriu et al. (2008), Mwangangi et al. (2007)
Anopheles cruciansGrieco et al. (2006), Qualls et al. (2012)
Anopheles domicolaDiagne et al. (1994)
Anopheles funestusGillies et al. (1968), Githeko et al. (1996)
Anopheles gambiaeCoggeshall (1944), Gillies et al. (1968), Huho et al. (2013)
Anopheles hyrcanusRueda et al. (2006, 2005), Ponçon et al., 2007), Aldemir et al. (2010)
Anopheles maculipennisAldemir et al. (2010), Brugman et al. (2015), Gordeev et al. (2005)
Anopheles meraukensisCooper et al. (2006), NSW Health (2016)
Anopheles paludisKarch and Mouchet (1992), Mouchet (1957)
Anopheles pharoensisGillies et al. (1968), Taye et al. (2006)
Anopheles philippinensisToma et al. (2002), Silver (2007), Bashar et al. (2012)
Anopheles pretoriensisAl-Sheik (2011), Shililu et al. (2003)
Anopheles punctipennisCarpenter and LaCasse (1974)
Anopheles quadrimaculatusCarpenter and LaCasse (1974)
Anopheles subpictusSinka et al. (2011)
Anopheles tesselatusMiyagi et al. (1983), Paramasivan et al. (2015)
Armigeres obturbansHarbach (2015)
Coquillettidia auritesSchwetz (1930), Njabo et al. (2009)
Coquillettidia linealisRussell et al. (2013), Williams (2005), Webb et al. (2016)
Coquillettidia metallicaNjabo et al. (2009), Mcclelland ga et al. (1960)
Coquillettidia perturbansCarpenter and LaCasse (1974), Anderson et al. (2007), Bosak et al. (2001), Callahan and Morris (1987)
Coquillettidia richiardiiVentim et al. (2012), Serandour et al. (2006), Versteirt et al. (2013)
Coquillettidia venezuelensisGuimarães et al. (2000), Degallier et al. (1978)
Culex adamesiSirivanakarn and Galindo (1980)
Culex annulirostrisNSW Health (2016), Hall-Mendelin et al. (2012), Williams and Kokkinn (2005)
Culex antennatusGad et al. (1995), Karch et al. (1993), Morsy et al. (1990), Kenawy et al. (1998)
Culex australicusNSW Health (2016), Russell (2012)
Culex bahamensisLopes (1997)
Culex bitaeniorhynchusKulkarni and Rajput (1988), Fakoorziba and Vijayan (2008), Harbach (1988)
Culex caudelliAlfonzo et al. (2005), Chadee and Tikasingh (1989)
Culex coronatorYee and Skiff (2014), de Oliveria et al. (1985)
Culex crybdade Oliveria et al. (1985)
Culex duttoniMwangangi et al. (2009)
Culex epidesmusKanojia (2003), Reisen et al. (1976)
Culex fatigansFlordia Medical Entomology Laboratory (2016), Liu et al. (1960), Robinson (2005)
Culex fuscocephalaOhba et al. (2015), Kulkarni and Rajput (1988), Amerasinghe and Munasingha (1994), Wang (1975)
Culex gelidusWilliams (2005), Sudeep (2014)
Culex guiartiLogan et al. (1991)
Culex modestusVeronesi et al. (2012), Radrova et al. (2013), Muñoz et al. (2012), Chalvet-Monfray et al. (2007), Fyodorova et al. (2006)
Culex nakuruensisSomeren (1967)
Culex neaveiDiallo et al. (2012a), Nikolay et al. (2012), Fall et al. (2013, 2011)
Culex nebulosusAdebote et al. (2006), Okorie (1978), Davis and Philip (1931)
Culex nigripalpusLaporta et al. (2008), Carpenter and LaCasse (1974), Flordia Medical Entomology Laboratory. (2016)
Culex p. molestusRobinson (2005), Gomes et al. (2013)
Culex perexiguusMuñoz et al. (2012), Ammar et al. (2012)
Culex perfuscusHopkins (1952), Diallo et al. (2014), Service (1993)
Culex pipiensHarbach (1988), Anderson et al. (2007)
Culex poicilipesMuturi et al. (2008), Yamar et al. (2005), Chevalier et al. (2004)
Culex pruinaWanson and Lebred (1946)
Culex pseudovishnuiFakoorziba and Vijayan (2008), Reisen et al. (1976), Amerasinghe and Indrajith (1995), Reuben et al. (1992)
Culex pullusJohansen et al. (2009), Webb et al. (2016)
Culex quinquefasciatusFlordia Medical Entomology Laboratory (2016), DeGroote and Sugumaran (2012)
Culex restuansApperson et al. (2002), Ebel et al. (2005), Kilpatrick et al. (2005), Molaei et al. (2008)
Culex rubinotusJupp et al. (1976)
Culex salinariusRochlin et al. (2008), Mackay et al. (2010), Rey et al. (2006)
Culex sitiensNSW Health (2016), Prummongkol et al. (2012)
Culex spissipesTakahashi (1968), Degallier et al. (1978)
Culex squamosesNSW Health (2016), Jansen et al. (2009)
Culex taeniopusDavies (1978), 1975), Lopes (1996)
Culex tarsalisReisen (1993), Rueger et al. (1964)
Culex telesillaNjogu and Kinoti (1971)
Culex thalassiusKerr (1932), Snow and Boreham (1978), Service (1993), Kirby et al. (2008)
Culex theileriAldemir et al. (2010), Muñoz et al. (2012), Simsek (2004)
Culex tritaeniorhynchusKanojia and Geevarghese (2004), Fakoorziba and Vijayan (2008), Flemings (1959), Amerasinghe and Munasingha (1994), Mwandawiro et al. (1999), Bhattacharyya et al. (1994), Reuben (1971)
Culex univittatusJupp (1967), Chandler et al. (1975), Jupp and Brown (1967)
Culex virgultusCarpenter and LaCasse (1974)
Culex vishnuiChen et al. (2014), Bhattacharyya et al. (1994), Ohba et al. (2015)
Culex vomeriferFerro et al. (2003), Natal et al. (1998), Suárez-Mutis et al. (2009), Sallum and Forattini (1996)
Culex wescheiSnow and Boreham (1973), Lane and Crosskey (2012)
Culex whitmoreiBegum et al. (1986), Reisen et al. (1976), Peiris et al. (1992)
Culex zombaensisLane and Crosskey (2012), Logan et al. (1991)
Culiseta alaskensisFrohne (1953)
Culiseta impatiensSommerman (1964), Frohne (1953), Murdock et al. (2010), Smith (1966)
Culiseta inornataCarpenter and LaCasse (1974), Smith (1966), Belton (1979)
Culiseta melanuraMolaei et al. (2006), Mahmood and Crans (1998), Flordia Medical Entomology Laboratory (2016), Hickman and Brown (2013)
Deinocerites pseudesMartin et al. (1973), Peyton et al. (1964)
Eretmapodites chrysogasterDoucet and Cachan (1961), Sylla et al. (2013), Service (1965a), Haddow (1946b)
Eretmapodites inornatusHaddow (1946a)
Eretmapodites oedipodeios (oedipodius)Haddow (1946a), de Cunha Ramos and Ribeiro (1990)
Eretmapodites quinquevittatusBohart and Ingram (1946), Jupp and Kemp (2002), Lounibos (1980)
Eretmapodites silvestrisLounibos (1980), Hoogstraal and Knight (1951)
Ficalbia flavensKing and Hoogstraal (1946)
Haemagogus anastasionisVan der Kuyp (1949), Bueno-Marà etal. (2015), Maestre-Serrano et al. (2013)
Haemagogus celesteBueno-Marà etal., 2015, Maestre-Serrano et al. (2013), Beran (1994), Chadee et al. (1985)
Haemagogus equinusChadee et al. (1985, 1993), Waddell and Taylor (1945)
Haemagogus janthinomysArnell (1973), Alencar et al. (2005), Chadee et al. (1992)
Haemagogus leucocelaenusAlencar et al. (2008), Pinto et al. (2009)
Haemagogus spegazziniiArnell (1973), Galindo et al. (1951, 1950)
Mansonia africanaKarch et al. (1993), Chandler et al. (1975), Hopkins (1952)
Mansonia septempunctataNSW Health (2016), Harbach (2015)
Mansonia titillansCarpenter and LaCasse (1974), Viana et al. (2010), Stein et al. (2013)
Mansonia uniformisSabesan et al. (1991), Kumar et al. (1989), Wharton (1962)
Mimomyia hispidaBoreham et al. (1975), Harbach (2015)
Mimomyia lacustrisHarbach (2015)
Mimomyia splendensBoreham et al. (1975), Robert et al. (1998)
Orthopodomyia signiferaHanson et al. (1995), Burkett-Cadena (2013)
Psorophora albipesAlfonzo et al. (2005), dos Santos Silva et al. (2012), Guimarães et al. (2000)
Psorophora columbiaeCarpenter and LaCasse (1974)
Psorophora feroxCarpenter and LaCasse (1974), Flordia Medical Entomology Laboratory (2016), Degallier et al. (1978), Molaei et al. (2008)
Runchomyia frontosaCardoso et al. (2015), Heinemann et al. (1980)
Sabethes albiprivusGomes et al. (2010), Pedro et al. (2008)
Sabethes belisarioiPinto et al. (2009)
Sabethes chloropterusBeran (1994), Pinto et al. (2009), Galindo (1958)
Sabethes soperiNavarro et al. (2015), Harbach (2015)
Uranotaenia mashonaensisHarbach and Schnur (2007)
Uranotaenia sapphirinaCupp et al. (2003), Crans (2016)
Uranotaenia unguiculataKhoshdel-Nezamiha et al. (2014), Ramsdale and Snow (2001), Sebesta et al. (2010), Bagirov et al. (1994), Kenawy et al. (1987)
Appendix 3—table 2

Primary sources for virus traits.

https://doi.org/10.7554/eLife.22053.013
VirusSources
Alfuy VirusMackenzie et al. (2012)
Bagaza virusMahy (2009), Llorente et al. (2015), Gamino et al. (2012)
Banzi virusGrard et al. (2010), Karabatsos (1985)
Bouboui virusGrard et al. (2010), Cook and Zumla (2009)
Bussuquara virusBeran (1994)
Dengue type 1Cook and Zumla (2009)
Dengue type 2Cook and Zumla (2009)
Dengue type 3Cook and Zumla (2009)
Dengue type 4Cook and Zumla (2009)
Edge Hill virusMackenzie et al. (2012), Doherty et al. (1964)
Iguape VirusCoimbra et al. (1993), Mahy (2009)
Ilheus virusMahy (2009), Chambers and Monath (2003), Laemmert and Hughes (1947), Aitken and Anderson (1959)
Israel turkey meningoencephalomyelitis virusMahy (2009), Nir (1972)
Japanese encephalitis virusMahy (2009), Burke and Leake (1988), Gresser et al. (1958)
Jugra virusNone
Kedougou virusCook and Zumla (2009), Diagne et al. (2015a)
Kokobera virusCook and Zumla (2009), Lequime and Lambrechts (2014)
Koutango virusChambers and Monath (2003), Cook and Zumla (2009)
Kunjin virusMahy (2009), Mackenzie et al. (2012)
Murray Valley encephalitis virusCook and Zumla (2009), Mackenzie et al. (2012)
Naranjal virusMahy (2009)
New Mapoon virusNisbet et al. (2005), Mahy (2009)
Ntaya virusMahy (2009)
Rocio virusMahy (2009), Cook and Zumla (2009)
Saboya virusMahy (2009), Traoré-Lamizana et al. (2001)
Sepik virusMackenzie et al. (2012), Cook and Zumla (2009)
Spondweni virusChambers and Monath (2003), Cook and Zumla (2009)
St. Louis encephalitis virusMackenzie et al. (2012), Cook and Zumla (2009)
Stratford virusMackenzie et al. (2012)
Tembusu virusMahy (2009), Tang et al. (2015)
Uganda S virusMahy (2009)
Usutu virusMahy (2009), Chambers and Monath (2003), Cook and Zumla (2009)
WesselbronMahy (2009), Chambers and Monath (2003), Cook and Zumla (2009)
West Nile virusMackenzie et al. (2012), Cook and Zumla (2009a), Mores et al. (2007), Turell et al. (2001)
Yaounde virusMackenzie et al. (2012)
Yellow fever virusMahy (2009)
Zika virusChambers and Monath (2003), Cook and Zumla (2009)

References

  1. 1
  2. 2
    Virus transmission studies with trinidadian mosquitoes II. further observations
    1. TH Aitken
    2. CR Anderson
    (1959)
    The American Journal of Tropical Medicine and Hygiene 8:41–45.
  3. 3
    Larval habitat, ecology, seasonal abundance and vectorial role in malaria transmission of anopheles arabiensis in Jazan region of saudi arabia
    1. AA Al-Sheik
    (2011)
    Journal of the Egyptian Society of Parasitology 41:615–634.
  4. 4
  5. 5
  6. 6
  7. 7
  8. 8
  9. 9
  10. 10
  11. 11
    Nocturnal biting rhythms of six mosquito species (Diptera: culicidae) in Kandy, Sri Lanka
    1. FP Amerasinghe
    2. NB Munasingha
    (1994)
    Journal of the National Science Council of Sri Lanka 22:279–290.
  12. 12
  13. 13
  14. 14
  15. 15
  16. 16
    Mosquito studies (DIPTERA, Culicidae) XXXII. a revision of the genus haemagogus
    1. JH Arnell
    (1973)
    Contributions of the American Entomological Institute 10:1–176.
  17. 17
    [The attack activity of Uranotaenia Unguiculata Edwards, 1913 mosquitoes on man]
    1. GA Bagirov
    2. EA Gadzhibekova
    3. GU Alirzaev
    (1994)
    Meditsinskaia Parazitologiia I Parazitarnye Bolezni 3:39–40.
  18. 18
  19. 19
    Proceedings of the 11th International Congress of Entomolgoy
    1. H Barnett
    (1960)
    341–345, The incrimination of arthropods as vectors of disease, Proceedings of the 11th International Congress of Entomolgoy, 1962.
  20. 20
  21. 21
  22. 22
    Mosquito populations diptera Culicidae of wetlands of an urban area
    1. G Becker
    2. D Neumann
    (1983)
    Zeitschrift Fuer Angewandte Zoologie 70:73–90.
  23. 23
    The ecology and seasonal fluctuations of mosquito larvae in a lake in Dhaka City bangladesh
    1. A Begum
    2. BR Biswas
    3. M Elias
    (1986)
    Bangladesh Journal of Zoology 14:41–48.
  24. 24
    The mosquitoes of Burnaby lake British-Columbia Canada
    1. P Belton
    (1979)
    Journal of the Entomological Society of British Columbia, 75.
  25. 25
    Variation in vector competence for dengue 2 virus among 24 collections of aedes aegypti from Mexico and the united states
    1. KE Bennett
    2. KE Olson
    3. ML Muñoz
    4. I Fernandez-Salas
    5. JA Farfan-Ale
    6. S Higgs
    7. WC Black
    8. BJ Beaty
    (2002)
    The American Journal of Tropical Medicine and Hygiene 67:85–92.
  26. 26
  27. 27
    Handbook of Zoonoses
    1. GW Beran
    (1994)
    Second Edition: Bacterial, Rickettsial, Chlamydial, and Mycotic Zoonoses, Handbook of Zoonoses, CRC Press.
  28. 28
    Host feeding patterns of culex vishnui sub group of mosquitoes in dibrugarh district of assam
    1. DR Bhattacharyya
    2. R Handique
    3. LP Dutta
    4. P Dutta
    5. P Doloi
    6. BK Goswami
    7. CK Sharma
    8. J Mahanta
    (1994)
    The Journal of Communicable Diseases 26:133–138.
  29. 29
    Mosquitoes of Okinawa and Islands in the Central Pacific
    1. RM Bohart
    2. RL Ingram
    (1946)
    US Dept of the Navy, Bureau of Medicine and Surgery.
  30. 30
  31. 31
  32. 32
  33. 33
    Habitat preference of host-seeking Coquillettidia perturbans (Walker) in relation to birds and eastern equine encephalomyelitis virus in new jersey
    1. PJ Bosak
    2. LM Reed
    3. WJ Crans
    (2001)
    Journal of Vector Ecology : Journal of the Society for Vector Ecology 26:103–109.
  34. 34
  35. 35
    Distribution of resting female aedes vexans (Meigen) in wooded and nonwooded areas of metropolitan Minneapolis-St. Paul, Minnesota
    1. CE Boxmeyer
    2. SM Palchick
    (1999)
    Journal of the American Mosquito Control Association 15:128–132.
  36. 36
  37. 37
  38. 38
    Emerging Zoonoses: Eco-Epidemiology, Involved Mechanisms and Public Health Implications
    1. R Bueno-MarÃ
    2. APG Almeida
    3. JC Navarro
    (2015)
    Frontiers Media SA.
  39. 39
    Japanese encephalitis
    1. DS Burke
    2. CJ Leake
    (1988)
    The Arboviruses: Epidemiology and Ecology 3:62–92.
  40. 40
    Mosquitoes of the Southeastern United States (1st edition)
    1. ND Burkett-Cadena
    (2013)
    Tuscaloosa: University of Alabama Press.
  41. 41
  42. 42
    Habitat characteristics of Coquillettidia perturbans in central florida
    1. JL Callahan
    2. CD Morris
    (1987)
    Journal of the American Mosquito Control Association 3:176–180.
  43. 43
  44. 44
  45. 45
  46. 46
    Mosquitoes of North America (North of Mexico)
    1. SJ Carpenter
    2. WJ LaCasse
    (1974)
    University of California Press.
  47. 47
  48. 48
  49. 49
    Distribution of haemagogus mosquitoes on small islands off Trinidad, W. I
    1. DD Chadee
    2. RC Persad
    3. N Andalcio
    4. W Ramdath
    (1985)
    Mosquito Systematics 17:147–153.
  50. 50
  51. 51
  52. 52
  53. 53
    The Flaviviruses: Detection, Diagnosis and Vaccine Development
    1. TJ Chambers
    2. TP Monath
    (2003)
    Academic Press.
  54. 54
  55. 55
  56. 56
  57. 57
  58. 58
  59. 59
  60. 60
  61. 61
    Anopheles gambiae in Brazil, 1930-40
    1. LT Coggeshall
    (1944)
    American Journal of Public Health and the Nations Health 34:75–76.
    https://doi.org/10.2105/AJPH.34.1.75
  62. 62
    Iguape: a newly recognized Flavivirus from São Paulo state, Brazil
    1. TL Coimbra
    2. ES Nassar
    3. AH Nagamori
    4. IB Ferreira
    5. LE Pereira
    6. IM Rocco
    7. M Ueda-Ito
    8. NS Romano
    (1993)
    Intervirology 36:144–152.
  63. 63
    Manson’s Tropical Diseases
    1. GC Cook
    2. A Zumla
    (2009)
    Elsevier Health Sciences.
  64. 64
  65. 65
  66. 66
    A note on the biting behaviour of the mosquito, aedes ochraceus, in a village in Kenya
    1. PS Corbet
    (1962)
    East African Medical Journal 39:511–514.
  67. 67
    The blood-feeding habits of aedes sollicitans (Walker) in relation to eastern equine encephalitis virus in coastal areas of new jersey. 2. results of experiments with caged mosquitoes and the effects of temperature and physiological age on host selection
    1. WJ Crans
    2. DA Sprenger
    3. F Mahmood
    (1996)
    Journal of Vector Ecology 21:1–5.
  68. 68
    The blood-feeding habits of Aedes sollicitans (Walker) in relation to eastern equine encephalitis virus in coastal areas of New Jersey. 3. Habitat preference, vertical distribution, and diel periodicity of host-seeking adults
    1. WJ Crans
    2. DA Sprenger
    (1996)
    Journal of Vector Ecology 21:6–13.
  69. 69
  70. 70
    Transmission of eastern equine encephalomyelitis virus in central Alabama
    1. EW Cupp
    2. K Klingler
    3. HK Hassan
    4. LM Viguers
    5. TR Unnasch
    (2003)
    The American Journal of Tropical Medicine and Hygiene 68:495–500.
  71. 71
    Identification and Geographical Distribution of the Mosquitos of North America, North of Mexico
    1. RF Darsie
    2. RA Ward
    (2005)
    University Press of Florida.
  72. 72
  73. 73
  74. 74
  75. 75
  76. 76
    Research on the mosquitoes of Angola XXI - Description of Eretmapodites angolensis sp. nov. and Eretmapodites dundo sp. nov. of the oedipodeios group
    1. H de Cunha Ramos
    2. H Ribeiro
    (1990)
    Garcia De Orta. Série De Zoologia 17:31–35.
  77. 77
    Alguns aspectos da ecologia dos mosquitos (Diptera: culicidae) de Uma area de planicie (Granjas Calabria), em Jacarepagua, rio de janeiro
    1. RL de Oliveria
    2. TF da Silva
    3. R Heyden
    (1985)
    II. Frequencia Mensal E No Ciclo Lunar. Memoirs Instituto Oswaldo Cruz-Fiocruz 80:123–133.
  78. 78
    Biting cycle of Culicidae in french guiana
    1. N Degallier
    2. F Pajot
    3. R Kramer
    4. J Claustre
    5. S Bellony
    (1978)
    Cahiers O.R.S.T.O.M. (Office De La Recherche Scientifique Et Technique Outre-Mer) Serie Entomologie Medicale Et Parasitologie 16:73–84.
  79. 79
  80. 80
    Mosquitoes feeding on brushtail possums (Trichosurus vulpecula) and humans in a native forest fragment in the Auckland region of New Zealand
    1. JG Derraik
    2. W Ji
    3. D Slaney
    (2007)
    The New Zealand Medical Journal 120:U2830.
  81. 81
  82. 82
    [Anopheles of Senegal. an annotated and illustrated list]
    1. N Diagne
    2. D Fontenille
    3. L Konate
    4. O Faye
    5. MT Lamizana
    6. F Legros
    7. JF Molez
    8. JF Trape
    (1994)
    Bulletin De La Société De Pathologie Exotique 87:267–277.
  83. 83
  84. 84
  85. 85
  86. 86
    Potential role of sylvatic and domestic african mosquito species in dengue emergence
    1. M Diallo
    2. AA Sall
    3. AC Moncayo
    4. Y Ba
    5. Z Fernandez
    6. D Ortiz
    7. LL Coffey
    8. C Mathiot
    9. RB Tesh
    10. SC Weaver
    (2005)
    The American Journal of Tropical Medicine and Hygiene 73:445–449.
  87. 87
    An arbovirus disease of present interest: yellow fever, its natural history facing an haemorragic fever, rift valley fever
    1. JP Digoutte
    (1999)
    Bulletin De La Société De Pathologie Exotique 92:343–348.
  88. 88
  89. 89
  90. 90
    Forest mosquitoes of the ivory coast republic. V. observations on the breeding places of mosquitoes of the genus Eretmapodites in the banco forest, Abidjan
    1. J Doucet
    2. P Cachan
    (1961)
    Bulletin De La Société De Pathologie Exotique 54:1253–1265.
  91. 91
  92. 92
  93. 93
  94. 94
  95. 95
  96. 96
  97. 97
    Breeding habitats of culex tritaeniorhynchus (Diptera: culicidae), A Japanese encephalitis vector, and associated mosquitoes in Mysore, India
    1. MR Fakoorziba
    2. A Vijayan
    (2008)
    Journal of the Entomological Research Society 10:1–9.
  98. 98
  99. 99
  100. 100
  101. 101
  102. 102
    The tree-hole species of mosquitoes of peiping, China
    1. LC Feng
    (1983)
    Chinese Medical Journal 2:503–525.
  103. 103
  104. 104
  105. 105
  106. 106
  107. 107
  108. 108
  109. 109
  110. 110
  111. 111
  112. 112
  113. 113
    Natural History of Culiseta Impatiens (Wlk.), (Diptera, Culicidae), in Alaska. Transactions of American Microscopical Society
    1. WC Frohne
    (1953)
    103–118, Natural History of Culiseta Impatiens (Wlk.), (Diptera, Culicidae), in Alaska. Transactions of American Microscopical Society, 72, 10.2307/3223507.
  114. 114
  115. 115
  116. 116
    Ecological observations on forest mosquitoes of an endemic yellow fever area in Panama
    1. P Galindo
    2. SJ Carpenter
    3. H Trapido
    (1951)
    The American Journal of Tropical Medicine and Hygiene 31:98–137.
  117. 117
    Observations on diurnal forest mosquitoes in relation to sylvan yellow fever in Panama
    1. P Galindo
    2. H Trapido
    3. SJ Carpenter
    (1950)
    The American Journal of Tropical Medicine and Hygiene 30:533–574.
  118. 118
    Bionomics of Sabethes chloropterus humboldt, a vector of sylvan yellow fever in middle america
    1. P Galindo
    (1958)
    The American Journal of Tropical Medicine and Hygiene 7:429–440.
  119. 119
  120. 120
    The aedes (Aedimorphus) Domesticus group (Diptera, Culicidae)
    1. B Geoffroy
    (1987)
    Mosquito Systematics 19:100–110.
  121. 121
    Isolation of the yellow fever virus from the aedes of the Aedes-Africanus group in the Central-African-Republic the importance of the humid and semi humid savanna in the emergence zone of the amaril virus
    1. M Germain
    2. P Sureau
    3. JP Herve
    4. J Fabre
    5. J Mouchet
    6. Y Robin
    7. B Geoffrey
    (1976)
    Cahiers O.R.S.T.O.M. (Office De La Recherche Scientifique Et Technique Outre-Mer) Serie Entomologie Medicale Et Parasitologie 14:125–140.
  122. 122
    Mosquito species composition, phenology and distribution (Diptera: culicidae) on prince Edward island
    1. DJ Giberson
    2. K Dau-Schmidt
    3. M Dobrin
    (2007)
    Journal of the Acadian Entomological Society 3:7–27.
  123. 123
  124. 124
    The anophelinae of Africa south of the sahara (Ethiopian zoogeographical region)
    1. MT Gillies
    2. B De Meillon
    (1968)
    Publications of the South African Institute for Medical Research 54:1–343.
  125. 125
  126. 126
    Ecologia de Haemagogus e Sabethes (Diptera: Culicidae) em áreas epizoóticas do vírus da febre amarela, Rio Grande do Sul, Brasil
    1. AdC Gomes
    2. MAN Torres
    3. MBd Paula
    4. A Fernandes
    5. AM Marassá
    6. CA Consales
    7. DF Fonseca
    (2010)
    Epidemiologia e Serviços de Saúde 19:101–113.
  127. 127
  128. 128
    Description of the new species anopheles artemievi sp.n
    1. M Gordeev I
    2. AB Zvantsov
    3. I Goriacheva I
    4. E ShaÄkevich V
    5. MN Ezhov
    (2005)
    Diptera, Culicidae). ResearchGate 2:4–5.
  129. 129
  130. 130
  131. 131
    Factors influencing transmission of japanese B encephalitis virus by a colonized strain of culex tritaeniorhynchus giles, from infected pigs and chicks to susceptible pigs and birds
    1. I Gresser
    2. JL Hardy
    3. SM Hu
    4. WF Scherer
    (1958)
    The American Journal of Tropical Medicine and Hygiene 7:365–373.
  132. 132
  133. 133
    Variation in susceptibility to oral infection with dengue viruses among geographic strains of aedes aegypti
    1. DJ Gubler
    2. S Nalim
    3. R Tan
    4. H Saipan
    5. J Sulianti Saroso
    (1979)
    The American Journal of Tropical Medicine and Hygiene 28:1045–1052.
  134. 134
  135. 135
  136. 136
  137. 137
    Twelve isolations of Zika virus from Aedes (Stegomyia) africanus (Theobald) taken in and above a Uganda forest
    1. AJ Haddow
    2. MC Williams
    3. JP Woodall
    4. DI Simpson
    5. LK Goma
    (1964)
    Bulletin of the World Health Organization 31:57–69.
  138. 138
  139. 139
  140. 140
  141. 141
  142. 142
  143. 143
  144. 144
  145. 145
  146. 146
  147. 147
    Notes on the biology of Orthopodomyia in Illinois
    1. SM Hanson
    2. RJ Novak
    3. RL Lampman
    4. MH Vodkin
    (1995)
    Journal of the American Mosquito Control Association 11:375–376.
  148. 148
  149. 149
    The mosquitoes of the subgenus culex in southwestern Asia and Egypt (Diptera: culicidae)
    1. RE Harbach
    (1988)
    ResearchGate 24:1–240.
  150. 150
  151. 151
  152. 152
  153. 153
    Changes in mosquito populations with expansion of the ross river reservoir, Australia, from stage 1 to stage 2A
    1. MN Hearnden
    2. BH Kay
    (1995)
    Journal of the American Mosquito Control Association 11:211–224.
  154. 154
    Collection records of the project "Mosquitoes of Middle America"
    1. SJ Heinemann
    2. THG Aitken
    3. JN Belkin
    (1980)
    Mosquito Systematics 12:179–284.
  155. 155
    Bioecologie comparee d’Aedes opok Corbet et Van Someren et A. africanus Theobald dans une galerie forestiere du sud de la Republique Centrafricaine
    1. JP Herve
    2. M Germain
    3. B Geoffroy
    (1975)
    Cah ORSTOM Ser Ent Med Et Parasitol 14:235–244.
  156. 156
    Influence de la clarte lunaire sur l’activite trophique d’Aedes taylori (Diptera, Culicidae)
    1. J Hervy
    2. F Legros
    3. L Ferrara
    (1986)
    Cah ORSTOM Ser Ent Med Et Parasitol 24:59–65.
  157. 157
    Culiseta Melanura Biology
    1. R Hickman
    2. J Brown
    (2013)
    Culiseta Melanura Biology.
  158. 158
    Observations on Eretmapodites silvestris Conchobius Edwards (Culicidae) in the Anglo-Egyptian Sudan
    1. H Hoogstraal
    2. K KNIGHT
    (1951)
    The American Journal of Tropical Medicine and Hygiene 31:659–664.
  159. 159
    Mosquitoes of the ethiopian region I. larval bionomics of mosquitoes and taxonomy of culicine larvae
    1. GHE Hopkins
    (1952)
    Adlard & Son, Ltd, British Museum of Natural History.
  160. 160
  161. 161
  162. 162
  163. 163
  164. 164
  165. 165
  166. 166
    The laboratory colonization of culex (Culex) univittatus theobald (Diptera: culicidae) from material collected in the highveld region of south Africa
    1. PG Jupp
    2. RG Brown
    (1967)
    Journal of the Entomological Society of Southern Africa 30:34–39.
  167. 167
    Studies on an outbreak of wesselsbron virus in the free state province, South Africa
    1. PG Jupp
    2. A Kemp
    (1998)
    Journal of the American Mosquito Control Association 14:40–45.
  168. 168
  169. 169
  170. 170
    A bionomic study of adult aedes (Neomelaniconion) circumluteolus in northern Kwazulu, South Africa
    1. PG Jupp
    2. BM McIntosh
    (1987)
    Journal of the American Mosquito Control Association 3:131–136.
  171. 171
    Larval habitats of culicine mosquitoes (Diptera: culcidae) in a sewage effluent disposal area in the south african high veld
    1. PG Jupp
    (1967)
    Journal of Entomology of South Africa 30:243–250.
  172. 172
  173. 173
  174. 174
    Bionomics of culex epidesmus associated with japanese encephalitis virus in India
    1. PC Kanojia
    (2003)
    Journal of the American Mosquito Control Association 19:151–154.
  175. 175
    International catalog of arboviruses including certain other viruses of vertebrates
    1. N Karabatsos
    (1985)
    The American Journal of Tropical Medicine and Hygiene 27:372–440.
  176. 176
    [The culicidian fauna and its nuisance in Kinshasha (Zaire)]
    1. S Karch
    2. N Asidi
    3. ZM Manzambi
    4. JJ Salaun
    (1993)
    Bulletin De La Société De Pathologie Exotique 86:68–75.
  177. 177
    [Anopheles paludis: important vector of malaria in zaire]
    1. S Karch
    2. J Mouchet
    (1992)
    Bulletin De La Société De Pathologie Exotique 85:388–389.
  178. 178
  179. 179
  180. 180
  181. 181
    Characterization of rice field mosquito habitats in Sharkia Governorate, Egypt
    1. MA Kenawy
    2. SS Rashed
    3. SS Teleb
    (1998)
    Journal of the Egyptian Society of Parasitology 28:449–459.
  182. 182
  183. 183
    Fauna and larval habitats of mosquitoes (Diptera: culicidae) of west Azerbaijan province, northwestern iran
    1. F Khoshdel-Nezamiha
    2. H Vatandoost
    3. S Azari-Hamidian
    4. MM Bavani
    5. F Dabiri
    6. R Entezar-Mahdi
    7. AR Chavshin
    (2014)
    Journal of Arthropod-Borne Diseases 8:163–173.
  184. 184
  185. 185
    Two new species of mosquitoes of the genus Ficalbia from netherlands new guinea
    1. WV King
    2. H Hoogstraal
    (1946)
    Proceedings of the Entomological Society of Washington 48:186–190.
  186. 186
  187. 187
  188. 188
    The aedes mosquitoes of the philippine islands
    1. KL Knight
    2. WB Hull
    (1953)
    Pacific Science 7:453–481.
  189. 189
  190. 190
    Day-Time resting habitats of culicine mosquitoes and their preponderance
    1. SM Kulkarni
    2. K Rajput
    (1988)
    Journal of Communicable Diseases 20:280–286.
  191. 191
    Biting rhythm of the vectors of malayan filariasis, mansonia annulifera, M. uniformis & M. indiana in shertallai (Kerala state), India
    1. NP Kumar
    2. S Sabesan
    3. KN Panicker
    (1989)
    The Indian Journal of Medical Research 89:52–55.
  192. 192
    Phylogeny of the genus Flavivirus
    1. G Kuno
    2. GJ Chang
    3. KR Tsuchiya
    4. N Karabatsos
    5. CB Cropp
    (1998)
    Journal of Virology 72:73–83.
  193. 193
    The virus of IlhéUs encephalitis; isolation, serological specificity and transmission. journal of immunology (Baltimore, md
    1. HW Laemmert
    2. TP Hughes
    (1947)
    1950) 55:61–67.
  194. 194
    Medical Insects and Arachnids
    1. RP Lane
    2. RW Crosskey
    (2012)
    Springer Science & Business Media.
  195. 195
  196. 196
    Description of the larva, pupa and female of aedes (N.) jemoti and revision of the keys to the subgenus neomelaniconion in Africa south of the Sahara
    1. R Le Berre
    2. J Hamon
    (1961)
    Bulletin De La Société De Pathologie Exotique 53:1054–1064.
  197. 197