Host reproductive phenology drives seasonal patterns of host use in mosquitoes

Seasonal shifts in host use by mosquitoes from birds to mammals drive the timing and intensity of annual epidemics of mosquito-borne viruses, such as West Nile virus, in North America. The biological mechanism underlying these shifts has been a matter of debate, with hypotheses falling into two camps: (1) the shift is driven by changes in host abundance, or (2) the shift is driven by seasonal changes in the foraging behavior of mosquitoes. Here we explored the idea that seasonal changes in host use by mosquitoes are driven by temporal patterns of host reproduction. We investigated the relationship between seasonal patterns of host use by mosquitoes and host reproductive phenology by examining a seven-year dataset of blood meal identifications from a site in Tuskegee National Forest, Alabama USA and data on reproduction from the most commonly utilized endothermic (white-tailed deer, great blue heron, yellow-crowned night heron) and ectothermic (frogs) hosts. Our analysis revealed that feeding on each host peaked during periods of reproductive activity. Specifically, mosquitoes utilized herons in the spring and early summer, during periods of peak nest occupancy, whereas deer were fed upon most during the late summer and fall, the period corresponding to the peak in births for deer. For frogs, however, feeding on early- and late-season breeders paralleled peaks in male vocalization. We demonstrate for the first time that seasonal patterns of host use by mosquitoes track the reproductive phenology of the hosts. Peaks in relative mosquito feeding on each host during reproductive phases are likely the result of increased tolerance and decreased vigilance to attacking mosquitoes by nestlings and brooding adults (avian hosts), quiescent young (avian and mammalian hosts), and mate-seeking males (frogs).     

Understanding host utilization by mosquitoes: determinants,challenges and future directions

Mosquito host utilization is a key factor in the transmission of vector-borne pathogens given that it greatly influences host–vector contact rates. Blood-feeding patterns of mosquitoes are not random, as some mosquitoes feed on particular species and/or individuals more than expected by chance. Mosquitoes use a number of cues including visual, olfactory, acoustic, and thermal stimuli emitted by vertebrate hosts to locate and identify their blood meal sources. Thus, differences in the quality/intensity of the released cues may drive host selection by mosquitoes at both inter- and intra-specific levels. Such patterns of host selection by mosquitoes in space and time can be structured by factors related to mosquitoes (e.g. innate host preference, behavioural plasticity), to hosts (e.g. emission of host-seeking cues, host availability) or to both (e.g. pathogen infection). In this study, we review current evidence, from phenomena to mechanisms, of how these factors influence host utilization by mosquitoes. We also review the methodologies commonly used in this research field and identify the major challenges for future studies. To bridge the knowledge gaps, we propose improvements to strengthen traditional approaches and the use of a functional trait-based approach to infer mosquito host utilization in natural communities.     

Frequency Dependence of Host Plant Choice within and Between Patches: A Large Cage Experiment

Oviposition preference is considered to be one of the most important factors behind patterns of host use among herbivorous insects. However, preference is defined as host plant choice under equal host abundance and availability, and it is likely that frequency-dependent effects will alter the actual pattern of host use beyond what preference trials reveals. The effects of such alterations are poorly known but could be important for the understanding of specialization and host shifts. We investigated how changes in frequency of a preferred and a less preferred host affected movement patterns and egg deposition within and among patches in a polyphagous butterfly, Polygonia c-album. Two experiments were carried out in large (8 × 30 m) outdoor cages, artificially divided into distinct patches with different frequencies of the two hosts: one that allowed for limited movement between patches and one that did not. There was a clear effect of frequency on patch selection; females spent more time in and laid more eggs in patches with a high frequency of the preferred host, which will potentially have a large effect on host use by modifying encounter rates in favor of the preferred host. However, there was no significant frequency-dependent plant choice within patches in any experiment. Instead, results indicate that females are distributing their eggs among plants species according to specific likelihoods of oviposition, independent of encounter rates, which is compatible with a strategy of risk-spreading.Co-ordinating editor: N. Yamamura     

Modelling trade-offs between mortality and fitness associated with persistent blood feeding by mosquitoes

Feeding persistence of mosquitoes (that tendency to attack again within a short period if a first meal is incomplete) is a key determinant of feeding success (defined here as blood-meal volume), especially with regard to mosquitoes that attack defensive hosts. Mortality risk from the vertebrate host is one cost that may be expected to create a trade-off against increased fecundity from second and subsequent meals depending on the availability of blood sources and on the probability of survival until the next gonotrophic cycle. We initially used stochastic simulation models to examine potential trade-offs in fecundity and survival for persistent and nonpersistent lines of mosquitoes that acquire blood from potentially dangerous hosts. In these models, a given strategy, i.e. persistence or the lack thereof, was fixed for the lifetime of the mosquito. Persistent blood feeding paid off relative to nonpersistent mosquitoes when feeding-associated mortality was relatively low to moderate and when the probability of obtaining blood on a given attempt was relatively low. On the other hand, the nonpersistent strategy tended to give relatively higher fecundity when the chances of obtaining blood were high. We also examined the implications of persistence as a feeding strategy for mosquitoes that could exhibit this behaviour in a flexible manner over their lifetime in response to state. We solved for optimal feeding behaviours with a stochastic dynamic programming model set up along similar lines to the simulation model. Results from this approach lend support to the idea that the threshold blood-meal size at which mosquitoes should continue to feed should change relative to the age of the mosquito. The modelling approaches used in this study identified important areas of future study for mosquito blood-feeding behaviour.     

Host patch selection induced by parasitism: basic reproduction ratio r(0) and optimal virulence

The effects of parasites on the behavior of their hosts are well documented. For example, parasites may affect the habitat selection of the host individual. We used variables aggregation methods to investigate the way in which parasites affect the spatial pattern of susceptible hosts. We developed a simple epidemiological model, taking into account both the reproduction processes of hosts (density-dependent birth and death) and infection, considered separately on two different patches, and the migration of susceptible hosts between these two patches. We used the complete model of three equations to generate an aggregated model describing the dynamics of the combined susceptible and infected host populations. We obtained the basic reproduction ratio (R(0)) from the aggregated model, and then studied the effect of the migratory behavior of susceptible hosts on the ability of the parasite to invade the system. We also used the basic reproduction ratio to investigate the evolution of parasite virulence in relation to the migration decisions of susceptible hosts. We found that host investment in avoidance of the infected patch leads to an increase in optimal virulence if host investment is costly.     

Fine-scale variation in vector host use and force of infection drive localized patterns of West Nile virus transmission

The influence of host diversity on multi-host pathogen transmission and persistence can be confounded by the large number of species and biological interactions that can characterize many transmission systems. For vector-borne pathogens, the composition of host communities has been hypothesized to affect transmission; however, the specific characteristics of host communities that affect transmission remain largely unknown. We tested the hypothesis that vector host use and force of infection (i.e., the summed number of infectious mosquitoes resulting from feeding upon each vertebrate host within a community of hosts), and not simply host diversity or richness, determine local infection rates of West Nile virus (WNV) in mosquito vectors. In suburban Chicago, Illinois, USA, we estimated community force of infection for West Nile virus using data on Culex pipiens mosquito host selection and WNV vertebrate reservoir competence for each host species in multiple residential and semi-natural study sites. We found host community force of infection interacted with avian diversity to influence WNV infection in Culex mosquitoes across the study area. Two avian species, the American robin (Turdus migratorius) and the house sparrow (Passer domesticus), produced 95.8% of the infectious Cx. pipiens mosquitoes and showed a significant positive association with WNV infection in Culex spp. mosquitoes. Therefore, indices of community structure, such as species diversity or richness, may not be reliable indicators of transmission risk at fine spatial scales in vector-borne disease systems. Rather, robust assessment of local transmission risk should incorporate heterogeneity in vector host feeding and variation in vertebrate reservoir competence at the spatial scale of vector-host interaction.     

Coevolution of Contrary Choices in Host-Parasitoid Systems

We investigate patch selection strategies of hosts and parasitoids in heterogeneous environments. Previous theoretical work showed that when host traits vary among patches, coevolved populations of hosts and parasitoids make congruent choices (i.e., hosts and parasitoids preferentially select the same patches) and exhibit direct density dependence in the distribution of percent parasitism. However, host-parasitoid systems in the field show a range of patterns in percent parasitism, while behavioral studies indicate that hosts and parasitoids can exhibit contrary choices (i.e., hosts avoid patches favored by the parasitoid). We extend previous theory by permitting life-history traits of the parasitoid as well as the host to vary among patches. Our analysis implies that in coevolutionarily stable populations, hosts preferentially select patches that intrinsically support higher host equilibrium numbers (i.e., the equilibrium number achieved by hosts when both populations are confined to a single patch) and that parasitoids preferentially select patches that intrinsically support higher parasitoid equilibrium numbers (i.e., the equilibrium number achieved by the parasitoids when both populations are confined to a patch). Using this result, we show how variation in life-history traits among patches leads to contrary or congruent choices or leads to direct density dependence, inverse density dependence, or density independence in the distribution of percent parasitism. In addition, we determine when populations playing the coevolutionarily stable strategies are ecologically stable. Our analysis shows that heterogeneous environments containing patches where the intrinsic rate of growth of the host and the survivorship rate of the parasitoid are low result in the coevolved populations exhibiting contrary choices and, as a result, promote ecological stability.     

Disease hotspots or hot species? Infection dynamics in multi‐host metacommunities controlled by species identity,not source location

Pathogen persistence in host communities is influenced by processes operating at the individual host to landscape‐level scale, but isolating the relative contributions of these processes is challenging. We developed theory to partition the influence of host species, habitat patches and landscape connectivity on pathogen persistence within metacommunities of hosts and pathogens. We used this framework to quantify the contributions of host species composition and habitat patch identity on the persistence of an amphibian pathogen across the landscape. By sampling over 11 000 hosts of six amphibian species, we found that a single host species could maintain the pathogen in 91% of observed metacommunities. Moreover, this dominant maintenance species contributed, on average, twice as much to landscape‐level pathogen persistence compared to the most influential source patch in a metacommunity. Our analysis demonstrates substantial inequality in how species and patches contribute to pathogen persistence, with important implications for targeted disease management.     

The oviposition behavior ofAphidius ervi andMonoctonus paulensis (Hymenoptera: Aphididae) encountering different host species (Homoptera: Aphididae) in sequential patches

Two-day-old mated females ofAphidius ervi Haliday andMonoctonus paulensis (Ashmead) were each provided with two sequential host patches. Patches were comprised of plastic petri dishes containing either 15 pea aphids,Acyrthosiphum pisum (Harris), or 15 alfalfa aphids,Macrosiphum creelii Davis. Both wasp species parasitized more hosts in patches containing pea aphids than in those containing alfalfa aphids, regardless of sequence. Females ofA. ervi also laid more eggs per aphid in patches containing pea aphids than in patches containing alfalfa aphids. When both patches contained alfalfa aphids,M. paulensis females parsitized more aphids in the second patch than in the first. Fewer alfalfa aphids were parasitized in the second patch when the first patch contained pea aphids, and fewer eggs were laid per alfalfa aphid. Parasitoid females of both species exhibited consistently higher rates of oviposition into their preferred host species and adjusted their reproductive allocation to hosts and host patches as a function of their experience in previous patches.     

Nestling discrimination without recognition: a possible defence mechanism for hosts towards cuckoo parasitism?

One of the great evolutionary puzzles is why hosts of parasitic birds discriminate finely against alien eggs, but almost never discriminate against parasitic chicks. A theoretical model has shown that an adaptive host response to alien eggs can be based on learning. However, learned nestling discrimination is too costly to be favoured by selection in hosts of evicting parasites, such as the European cuckoo (Cuculus canorus). Indeed, parasitic chick rejection has never been reported for any European cuckoo host species. As learned nestling discrimination is maladaptive, one can expect that a viable alternative for hosts would be to use discrimination mechanisms not involving learning and/or recognition. We suggest that hosts may starve and desert cuckoo chicks that require higher amounts of food than an average host brood at fledging (i.e. feeding rates to a parasite are outside the normal range of host behaviour in unparasitized nests). Our observations of the reed warbler (Acrocephalus scirpaceus) at parasitized nests indicate that such behaviour could possibly work in this host species.     

Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing

Understanding mosquito host choice is important for assessing vector competence or identifying disease reservoirs. Unfortunately, the availability of an unbiased method for comprehensively evaluating the composition of insect blood meals is very limited, as most current molecular assays only test for the presence of a few pre-selected species. These approaches also have limited ability to identify the presence of multiple mammalian hosts in a single blood meal. Here, we describe a novel high-throughput sequencing method that enables analysis of 96 mosquitoes simultaneously and provides a comprehensive and quantitative perspective on the composition of each blood meal. We validated in silico that universal primers targeting the mammalian mitochondrial 16S ribosomal RNA genes (16S rRNA) should amplify more than 95% of the mammalian 16S rRNA sequences present in the NCBI nucleotide database. We applied this method to 442 female Anopheles punctulatus s. l. mosquitoes collected in Papua New Guinea (PNG). While human (52.9%), dog (15.8%) and pig (29.2%) were the most common hosts identified in our study, we also detected DNA from mice, one marsupial species and two bat species. Our analyses also revealed that 16.3% of the mosquitoes fed on more than one host. Analysis of the human mitochondrial hypervariable region I in 102 human blood meals showed that 5 (4.9%) of the mosquitoes unambiguously fed on more than one person. Overall, analysis of PNG mosquitoes illustrates the potential of this approach to identify unsuspected hosts and characterize mixed blood meals, and shows how this approach can be adapted to evaluate inter-individual variations among human blood meals. Furthermore, this approach can be applied to any disease-transmitting arthropod and can be easily customized to investigate non-mammalian host sources.     

Divide and conquer? Persistence of infectious agents in spatial metapopulations of hosts

Persistence of an infectious agent in a population is an important issue in epidemiology. It is assumed that spatially fragmenting a population of hosts increases the probability of persistence of an infectious agent and that movement of hosts between the patches is vital for that. The influence of migration on persistence is however often studied in mean-field models, whereas in reality the actual distance travelled can be limited and influence the movement dynamics. We use a stochastic model, where within- and between-patch dynamics are coupled and movement is modelled explicitly, to show that explicit consideration of movement distance makes the relation between persistence of infectious agents and the metapopulation structure of its hosts less straightforward than previously thought. We show that the probability of persistence is largest at an intermediate movement distance of the host and that spatially fragmenting a population of hosts is not necessarily beneficial for persistence.     

Host patch traits have scale-dependent effects on diversity in a stickleback parasite metacommunity

Many metacommunities are distributed across habitat patches that are themselves aggregated into groups. Perhaps the clearest example of this nested metacommunity structure comes from multi-species parasite assemblages, which occupy individual hosts that are aggregated into host populations. At both spatial scales, we expect parasite community diversity in a given patch (either individual host or population) to depend on patch characteristics that affect colonization rates and species sorting. But, are these patch effects consistent across spatial scales? Or, do different processes govern the distribution of parasite community diversity among individual hosts, versus among host patches? To answer these questions, we document the distribution of parasite richness among host individuals and among populations in a metapopulation of threespine stickleback Gasterosteus aculeatus. We find some host traits (host size, gape width) are associated with increased parasite richness at both spatial scales. Other patch characteristics affect parasite richness only among individuals (sex), or among populations (lake size, lake area, elevation and population mean heterozygosity). These results demonstrate that some rules governing parasite richness in this metacommunity are shared across scales, while others are scale-specific.     

The impact of infection on host competition and its relationship to parasite persistence in a Daphnia microparasite system

Evolutionary studies often estimate fitness components with the aim to make predictions about the outcome of selection. Depending on the system and the question, different fitness components are used, but their usefulness for predicting the outcome of selection is rarely tested. Here we estimate host fitness components in different ways with the aim to test how well they agree with each other and how well they predict host fitness at the population level in the presence of the parasite. We use a Daphnia magna-microparasite system to study the competitive ability of host clones in the absence and presence of the parasite, the infection intensity of the parasite in individuals of twelve host clones (an estimate of both host resistance and parasite reproductive success), and parasite persistence in small host populations (an estimate of R ₀ of the parasite). Analysis of host competitive ability and parasite persistence reveals strong host genotype effects, while none are found for infection intensity. Host competitive ability further shows a genotype-specific change upon infection, which is correlated with the relative persistence of the parasite in the competing hosts. Hosts in which the parasite persists better suffer a competitive disadvantage in the parasite’s presence. This suggests that in this system, parasite-mediated selection can be predicted by parasite persistence, but not by parasite infection intensity.     

Evolution of host specificity drives reproductive isolation among RNA viruses

Ecological speciation hypotheses claim that assortative mating evolves as a consequence of divergent natural selection for ecologically important traits. Reproductive isolation is expected to be particularly likely to evolve by this mechanism in species such as phytophagous insects that mate in the habitats in which they eat. We tested this expectation by monitoring the evolution of reproductive isolation in laboratory populations of an RNA virus that undergoes genetic exchange only when multiple virus genotypes coinfect the same host. We subjected four populations of the RNA bacteriophage phi6 to 150 generations of natural selection on a novel host. Although there was no direct selection acting on host range in our experiment, three of the four populations lost the ability to infect one or more alternative hosts. In the most extreme case, one of the populations evolved a host range that does not contain any of the hosts infectible by the wild-type phi6. Whole genome sequencing confirmed that the resulting reproductive isolation was due to a single nucleotide change, highlighting the ease with which an emerging RNA virus can decouple its evolutionary fate from that of its ancestor. Our results uniquely demonstrate the evolution of reproductive isolation in allopatric experimental populations. Furthermore, our data confirm the biological credibility of simple "no-gene" mechanisms of assortative mating, in which this trait arises as a pleiotropic effect of genes responsible for ecological adaptation.     

The effect of Jacobin Cuckoo Clamator jacobinus parasitism on the body mass and survival of young in a new host species

The southern African subspecies of Jacobin Cuckoo Clamator jacobinus serratus is a brood parasite of a range of host species. While Jacobin Cuckoos do not evict host young, previous research has found that host young rarely survive the nestling period. Here we provide the first records of Jacobin Cuckoo parasitism of a new host species, the Southern Pied Babbler Turdoides bicolor. We investigate rates of brood parasitism and the survival of host young. The Southern Pied Babbler is one of the largest recorded hosts for Jacobin Cuckoos and, unusually, we find that host young tend to survive the nestling period and maintain similar body mass to host young in unparasitized broods. However, host young were less likely to survive to independence than young raised in unparasitized nests, suggesting a post‐fledging reproductive cost to hosts.     

Differential reproductive success favours strong host preference in a highly specialized brood parasite

Obligate avian brood parasites show dramatic variation in the degree to which they are host specialists or host generalists. The screaming cowbird Molothrus rufoaxillaris is one of the most specialized brood parasites, using a single host, the bay-winged cowbird (Agelaioides badius) over most of its range. Coevolutionary theory predicts increasing host specificity the longer the parasite interacts with a particular avian community, as hosts evolve defences that the parasite cannot counteract. According to this view, host specificity can be maintained if screaming cowbirds avoid parasitizing potentially suitable hosts that have developed effective defences against parasitic females or eggs. Specialization may also be favoured, even in the absence of host defences, if the parasite's reproductive success in alternative hosts is lower than that in the main host. We experimentally tested these hypotheses using as alternative hosts two suitable but unparasitized species: house wrens (Troglodytes aedon) and chalk-browed mockingbirds (Mimus saturninus). We assessed host defences against parasitic females and eggs, and reproductive success of the parasite in current and alternative hosts. Alternative hosts did not discriminate against screaming cowbird females or eggs. Egg survival and hatching success were similarly high in current and alternative hosts, but the survival of parasitic chicks was significantly lower in alternative hosts. Our results indicate that screaming cowbirds have the potential to colonize novel hosts, but higher reproductive success in the current host may favour host fidelity.     

Winter severity predicts the timing of host shifts in the mosquito Culex erraticus

In temperate regions, seasonal epidemics of many mosquito-borne viruses are triggered when mosquito populations shift from feeding on avian to mammalian hosts. We investigated effects of temperature on the timing of bird-to-mammal shifts using an 8 year dataset of blood-meals from a mosquito (Culex erraticus) in Alabama, USA. As expected, Cx. erraticus shifted from avian to mammalian hosts each year. The timing of the shift, however, varied considerably among years. Harshness of the preceding winter (chill accumulation) explained 93 per cent of the variation in the timing of bird-to-mammal shifts, with shifts occurring later in years following harsher winters. We hypothesize that winter temperatures drive the timing of bird-to-mammal shifts through effects on host reproductive phenology. Because mosquitoes target birds during the nesting season, and bird nesting occurs later in years following colder winters, later nesting dates result in a concomitant delay in the timing of bird-to-mammal host shifts. Global increases in winter temperatures could cause significant changes in the timing of seasonal host shifts by mosquitoes, with prolonged periods of epidemic transmission of mosquito-borne diseases.     

Inference of parasite local adaptation using two different fitness components

Estimating parasite fitness is central to studies aiming to understand parasite evolution. Theoretical models generally use the basic reproductive rate R(0) to express fitness, yet it is very difficult to quantify R(0) empirically and experimental studies often use fitness components such as infection intensity or infectivity as substitutes. These surrogate measures may be biased in several ways. We assessed local adaptation of the microsporidium Ordospora colligata to its host, the crustacean Daphnia magna using two different parasite fitness components: infection persistence over several host generations in experimental populations and infection intensity in individual hosts. We argue that infection persistence is a close estimator of R(0), whereas infection intensity measures only a component of it. Both measures show a pattern that is consistent with parasite local adaptation and they correlate positively. However, several inconsistencies between them suggest that infection intensity may at times provide an inadequate estimate of parasite fitness.     

West Nile virus experimental evolution in vivo and the trade-off hypothesis

In nature, arthropod-borne viruses (arboviruses) perpetuate through alternating replication in vertebrate and invertebrate hosts. The trade-off hypothesis proposes that these viruses maintain adequate replicative fitness in two disparate hosts in exchange for superior fitness in one host. Releasing the virus from the constraints of a two-host cycle should thus facilitate adaptation to a single host. This theory has been addressed in a variety of systems, but remains poorly understood. We sought to determine the fitness implications of alternating host replication for West Nile virus (WNV) using an in vivo model system. Previously, WNV was serially or alternately passed 20 times in vivo in chicks or mosquitoes and resulting viruses were characterized genetically. In this study, these test viruses were competed in vivo in fitness assays against an unpassed marked reference virus. Fitness was assayed in chicks and in two important WNV vectors, Culex pipiens and Culex quinquefasciatus. Chick-specialized virus displayed clear fitness gains in chicks and in Cx. pipiens but not in Cx. quinquefasciatus. Cx. pipiens-specialized virus experienced reduced fitness in chicks and little change in either mosquito species. These data suggest that when fitness is measured in birds the trade-off hypothesis is supported; but in mosquitoes it is not. Overall, these results suggest that WNV evolution is driven by alternate cycles of genetic expansion in mosquitoes, where purifying selection is weak and genetic diversity generated, and restriction in birds, where purifying selection is strong.