Insights into Arbovirus Evolution and Adaptation from Experimental Studies

Arthropod-borne viruses (arboviruses) are maintained in nature by cycling between vertebrate hosts and haematophagous invertebrate vectors. These viruses are responsible for causing a significant public health burden throughout the world, with over 100 species having the capacity to cause human disease. Arbovirus outbreaks in previously naïve environments demonstrate the potential of these pathogens for expansion and emergence, possibly exacerbated more recently by changing climates. These recent outbreaks, together with the continued devastation caused by endemic viruses, such as Dengue virus which persists in many areas, demonstrate the need to better understand the selective pressures that shape arbovirus evolution. Specifically, a comprehensive understanding of host-virus interactions and how they shape both host-specific and virus-specific evolutionary pressures is needed to fully evaluate the factors that govern the potential for host shifts and geographic expansions. One approach to advance our understanding of the factors influencing arbovirus evolution in nature is the use of experimental studies in the laboratory. Here, we review the contributions that laboratory passage and experimental infection studies have made to the field of arbovirus adaptation and evolution, and how these studies contribute to the overall field of arbovirus evolution. In particular, this review focuses on the areas of evolutionary constraints and mutant swarm dynamics; how experimental results compare to theoretical predictions; the importance of arbovirus ecology in shaping viral swarms; and how current knowledge should guide future questions relevant to understanding arbovirus evolution

Geographic variation in the response of Culex pipiens life history traits to temperature

BackgroundClimate change is predicted to alter the transmission of many vector-borne pathogens. The quantitative impact of climate change is usually estimated by measuring the temperature-performance relationships for a single population of vectors, and then mapping this relationship across a range of temperatures or locations. However, life history traits of different populations often differ significantly. Specifically, performance across a range of temperatures is likely to vary due to local adaptation to temperature and other factors. This variation can cause spatial variation in pathogen transmission and will influence the impact of climate change on the transmission of vector-borne pathogens.MethodsWe quantified variation in life history traits for four populations of Culex pipiens (Linnaeus) mosquitoes. The populations were distributed along altitudinal and latitudinal gradients in the eastern United States that spanned ~3 °C in mean summer temperature, which is similar to the magnitude of global warming expected in the next 3-5 decades. We measured larval and adult survival, development rate, and biting rate at six temperatures between 16 and 35 °C, in a common garden experiment.ResultsTemperature had strong and consistent non-linear effects on all four life history traits for all four populations. Adult female development time decreased monotonically with increasing temperature, with the largest decrease at cold temperatures. Daily juvenile and adult female survival also decreased with increasing temperature, but the largest decrease occurred at higher temperatures. There was significant among-population variation in the thermal response curves for the four life history traits across the four populations, with larval survival, adult survival, and development rate varying up to 45, 79, and 84 % among populations, respectively. However, variation was not correlated with local temperatures and thus did not support the local thermal adaptation hypothesis.ConclusionThese results suggest that the impact of climate change on vector-borne disease will be more variable than previous predictions, and our data provide an estimate of this uncertainty. In addition, the variation among populations that we observed will shape the response of vectors to changing climates

West Nile virus envelope protein glycosylation is required for efficient viral transmission by Culex vectors

AbstractMany, but not all, strains of West Nile virus (WNV) contain a single N-linked glycosylation site on their envelope (E) proteins. Previous studies have shown that E-glycosylated strains are more neuroinvasive in mice than non-glycosylated strains. E protein glycosylation also appears to play a role in attachment and entry of WNV into host cells in vitro; however, studies examining how E protein glycosylation affects the interactions of WNV with its mosquito vectors in vivo have not yet been performed. We mutated the E protein glycosylation site from NYS to IYS in a previously described full-length clone of the NY99 genotype of WNV (WT), resulting in a virus that lacked the glycan at aa154. WNV-N154I replicated less efficiently than WNV-WT in Culex mosquito tissues, although the extent of the decrease was greater in Cx. pipiens than in Cx. tarsalis. Following peroral infection, mosquitoes infected with WNV-N154I were less likely to transmit virus than those infected with WNV-WT. Interestingly, all but one of the mosquitoes infected with WNV-N154I transmitted a revertant virus, suggesting that there is strong selective pressure toward E protein glycosylation. Together these data suggest that loss of the glycan at aa154 on the WNV E protein can severely restrict viral spread in the mosquito vector

The costs of infection and resistance as determinants of West Nile virus susceptibility in Culex mosquitoes

<p>Abstract</p> <p>Background</p> <p>Understanding the phenotypic consequences of interactions between arthropod-borne viruses (arboviruses) and their mosquito hosts has direct implications for predicting the evolution of these relationships and the potential for changes in epidemiological patterns. Although arboviruses are generally not highly pathogenic to mosquitoes, pathology has at times been noted. Here, in order to evaluate the potential costs of <it>West Nile virus </it>(WNV) infection and resistance in a primary WNV vector, and to assess the extent to which virus-vector relationships are species-specific, we performed fitness studies with and without WNV exposure using a highly susceptible <it>Culex pipiens </it>mosquito colony. Specifically, we measured and compared survival, fecundity, and feeding rates in bloodfed mosquitoes that were (i) infected following WNV exposure (susceptible), (ii) uninfected following WNV exposure (resistant), or (iii) unexposed.</p> <p>Results</p> <p>In contrast to our previous findings with a relatively resistant <it>Cx. tarsalis </it>colony, WNV infection did not alter fecundity or blood-feeding behaviour of <it>Cx. pipiens</it>, yet results do indicate that resistance to infection is associated with a fitness cost in terms of mosquito survival.</p> <p>Conclusions</p> <p>The identification of species-specific differences provides an evolutionary explanation for variability in vector susceptibility to arboviruses and suggests that understanding the costs of infection and resistance are important factors in determining the potential competence of vector populations for arboviruses.</p

Cooperative interactions in the West Nile virus mutant swarm

<p>Abstract</p> <p>Background</p> <p>RNA viruses including arthropod-borne viruses (arboviruses) exist as highly genetically diverse mutant swarms within individual hosts. A more complete understanding of the phenotypic correlates of these diverse swarms is needed in order to equate RNA swarm breadth and composition to specific adaptive and evolutionary outcomes.</p> <p>Results</p> <p>Here, we determined clonal fitness landscapes of mosquito cell-adapted <it>West Nile virus</it> (WNV) and assessed how altering the capacity for interactions among variants affects mutant swarm dynamics and swarm fitness. Our results demonstrate that although there is significant mutational robustness in the WNV swarm, genetic diversity also corresponds to substantial phenotypic diversity in terms of relative fitness <it>in vitro</it>. In addition, our data demonstrate that increasing levels of co-infection can lead to widespread strain complementation, which acts to maintain high levels of phenotypic and genetic diversity and potentially slow selection for individual variants. Lastly, we show that cooperative interactions may lead to swarm fitness levels which exceed the relative fitness levels of any individual genotype.</p> <p>Conclusions</p> <p>These studies demonstrate the profound effects variant interactions can have on arbovirus evolution and adaptation, and provide a baseline by which to study the impact of this phenomenon in natural systems.</p

Serologic Evidence of West Nile Virus Transmission, Jamaica, West Indies

In spring 2002, an intensive avian serosurvey was initiated in Jamaica, Puerto Rico, and Mexico. We collected >1,600 specimens from resident and nonresident neotropical migratory birds before their northerly migrations. Plaque reduction neutralization test results indicated specific neutralizing antibodies to West Nile virus in 11 resident species from Jamaica

Vector Competence of Argentine Mosquitoes (Diptera: Culicidae) for West Nile virus (Flaviviridae: Flavivirus)

We examined the ability of Culex pipiens L. complex mosquitoes from Argentina to vector West Nile virus (WNV) to assess their role in the transmission of WNV in South America. Several egg rafts of Culex spp. were collected from different breeding sites in the suburbs of the city of La Plata, Argentina, and a subset of each progeny was scored with morphological and genetic species indicators. Surprisingly, we did not find Cx. pipiens form pipiens, but found evidence of genetic hybrids of Culex quinquefasciatus and Cx. pipiens f. molestus. We then used morphological traits to create two colonies predominantly composed of one of these two taxa, although some hybrids are likely to have been included in both. These colonies were used in vector competence studies using NY99 and WN02 genotype strains of WNV obtained in New York State. As controls, we also tested colonies of U.S. Cx. quinquefasciatus and Cx. pipiens f. molestus. Additional Culex larvae from three drainage ditches near the cities of La Plata and Berisso, Argentina, were identified by morphological and high-resolution molecular markers (microsatellites) as Cx. quinquefasciatus Say, Cx. pipiens form molestus, and hybrids. Results indicate that Argentinian Culex are competent but only moderately efficient vectors of WNV and are less susceptible to this virus than comparable U.S. mosquito strains. Studies of vertical transmission of NY99 virus by Cx. pipiens f. molestus hybrids from Argentina yielded a minimal filial infection rate of 1.19 from females feeding during their second and later bloodmeals.Centro de Estudios Parasitológicos y de Vectore

Characterization of Rabensburg Virus, a Flavivirus Closely Related to West Nile Virus of the Japanese Encephalitis Antigenic Group

Rabensburg virus (RABV), a Flavivirus with ∼76% nucleotide and 90% amino acid identity with representative members of lineage one and two West Nile virus (WNV), previously was isolated from Culex pipiens and Aedes rossicus mosquitoes in the Czech Republic, and phylogenetic and serologic analyses demonstrated that it was likely a new lineage of WNV. However, no direct link between RABV and human disease has been definitively established and the extent to which RABV utilizes the typical WNV transmission cycle is unknown. Herein, we evaluated vector competence and capacity for vertical transmission (VT) in Cx. pipiens; in vitro growth on avian, mammalian, and mosquito cells; and infectivity and viremia production in birds. RABV infection and replication only were detected on mosquito cells. Experimentally inoculated birds did not become infected. Cx. pipiens had poor peroral vector competence and a higher VT rate as compared to US-WNV in Cx. pipiens. As a result, we postulate that RABV is an intermediate between the mosquito-specific and horizontally transmitted flaviviruses

Factors Related to Aedes aegypti (Diptera: Culicidae) Populations and Temperature Determine Differences on Life-History Traits With Regional Implications in Disease Transmission

Aedes aegypti (L.) (Diptera: Culicidae) is a vector of many medically significant viruses in the Americas, including dengue virus, chikungunya virus, and Zika virus. Traits such as longevity, fecundity, and feeding behavior contribute to the ability of Ae. aegypti to serve as a vector of these pathogens. Both local environmental factors and population genetics could contribute to variability in these traits. We performed a comparative study of Ae. aegypti populations from four geographically and environmentally distinct collection sites in Argentina in which the cohorts from each population were held at temperature values simulating a daily cycle, with an average of 25°C in order to identify the influence of population on life-history traits. In addition, we performed the study of the same populations held at a daily temperature cycle similar to that of the surveyed areas. According to the results, Aguaray is the most outstanding population, showing features that are important to achieve high fitness. Whereas La Plata gathers features consistent with low fitness. Iguazu was outstanding in blood-feeding rate while Posadas's population showed intermediate values. Our results also demonstrate that climate change could differentially affect unique populations, and that these differences have implications for the capacity for Ae. aegypti to act as vectors for medically important arboviruses.Fil: Muttis, Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Balsalobre, Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Chuchuy, Ailen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Mangudo, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones en Energía no Convencional. Universidad Nacional de Salta. Facultad de Ciencias Exactas. Departamento de Física. Instituto de Investigaciones en Energía no Convencional; ArgentinaFil: Ciota, Alexander. Wadsworth Center, New York State Department Of Health; Estados UnidosFil: Micieli, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentin