Ectoparasitic mites exert non-consumptive effects on the larvae of a fruit fly host.

The mere presence of predators or parasites can negatively impact the fitness of prey or hosts. Exposure to predators during an organism's development can have deleterious effects on juvenile survival and the subsequent adult stage. Currently, it is unknown if parasites have analogous impacts on host larval stages and whether these effects carry over into other subsequent life stages. However, parasites may be exerting widespread yet underestimated non-consumptive effects (NCEs). We tested if Drosophila nigrospiracula larvae avoid pupating near mite cues (caged Macrocheles subbadius) in arena experiments, and measured the rate of pupation in arenas with mites and arenas without mites. Larvae disproportionately pupated on the side of arenas that lacked mite cues. Furthermore, fewer larvae successfully pupated in arenas containing mites cues compared to arenas without mite cues. We found that ectoparasitic mites exert NCEs on Drosophila larvae, even though the larval stage is not susceptible to infection. We discuss these results in the context of parasite impacts on host population growth in an infectious world.

Parasite preferences for large host body size can drive overdispersion in a fly-mite association.

Body size generally correlates intraspecifically with insect fitness but can also correlate with parasite abundance (number of parasites). Host preferences by parasites, and variation in host immunity, could contribute to this trend. We investigated the effect of host size on mite-fly interactions (Macrocheles subbadius and Drosophila nigrospiracula). Mites strongly preferred to infect larger flies in pair-wise choices, and larger flies were more likely to be infected and acquired more mites in infection microcosms. Preferences of parasites resulted in size-biased infection outcomes. We discuss the implications of this heterogeneity in infection on parasite overdispersion and fly populations.

Relative contributions of parasite consumptive and non-consumptive effects to host population suppression in simulated fly-mite populations.

The "ecology of fear" framework was developed to describe the impacts predators have on potential prey and prey populations, outside of consumption/predation (i.e. non-consumptive effects, NCEs). This framework has recently been extended to symbiotic interactions such as host-parasite associations. Although the NCEs of predators and parasites on their individual victims can be measured experimentally, it is currently not known whether parasites can exert population-level effects on potential hosts through their NCEs. Modelling can be a useful tool for scaling individual-level NCEs to populations to determine impacts on host population growth. In this study, we used previously published data on the consumptive and non-consumptive effects of an ectoparasitic mite (Macrocheles subbadius) on a fruit fly (Drosophila nigrospiracula) to simulate populations experiencing fear (NCEs only), both fear and infection (consumption + NCEs) or neither. Population-level models indicate that NCEs alone were insufficient to reduce population growth. In fact, host populations experiencing NCEs but not infection had slightly larger final populations than unexposed populations (by ~ 550 flies). This result suggests there is compensation (i.e. increased daily reproduction that overcomes shorter lifespans) among exposed flies. By contrast, the consumptive effects of parasites suppressed the growth of simulated host populations, and this deleterious impact grew non-linearly with infection prevalence.

Scared of the dark? Phototaxis as behavioural immunity in a host-parasite system.

Behavioural immunity describes suites of behaviours hosts use to minimize the risks of infection by parasites/pathogens. Research has focused primarily on the evasion and physical removal of infectious stages, as well as behavioural fever. However, other behaviours affect infection risk while carrying ecologically significant trade-offs. Phototaxis, in particular, has host fitness implications (e.g. altering feeding and thermoregulation) that also impact infection outcomes. In this study, we hypothesized that a fly host, Drosophila nigrospiracula, employs phototaxis as a form of behavioural immunity to reduce the risk of infection. First, we determined that the risk of infection is lower for flies exposed in the light relative to the dark using micro-arena experiments. Because Drosophila vary in ectoparasite resistance based on mating status we examined parasite-mediated phototaxis in mated and unmated females. We found that female flies spent more time in the light side of phototaxis chambers when mites were present than in the absence of mites. Mating marginally decreased female photophobia independently of mite exposure. Female flies moved to lighter, i.e. less infectious, environments when threatened with mites, suggesting phototaxis is a mechanism of behavioural immunity. We discuss how parasite-mediated phototaxis potentially trades-off with host nutrition and thermoregulation.

Endosymbiotic Male-Killing Spiroplasma Affects the Physiological and Behavioral Ecology of Macrocheles-Drosophila Interactions.

While many arthropod endosymbionts are vertically transmitted, phylogenetic studies reveal repeated introductions of hemolymph-dwelling Spiroplasma into Drosophila. Introductions are often attributed to horizontal transmission via ectoparasite vectors. Here, we test if mites (Macrocheles subbadius) prefer to infect Spiroplasma poulsonii MSRO (Melanogaster sex ratio organism)-infected flies and if MSRO infection impairs fly resistance against secondary mite attack. First, we tested if mites prefer MSRO+ or MSRO- flies using pairwise choice tests across fly ages. We then tested whether mite preferences are explained by changes in fly physiology, specifically increased metabolic rate (measured as CO2 production). We hypothesize that this preference is due in part to MSRO+ flies expressing higher metabolic rates. However, our results showed mite preference depended on an interaction between fly age and MSRO status: mites avoided 14-day-old MSRO+ flies relative to MSRO- flies (31% infection) but preferred MSRO+ flies (64% infection) among 26-day-old flies. Using flowthrough respirometry, we found 14-day-old MSRO+ flies had higher CO2 emissions than MSRO- flies (32% greater), whereas at 26 days old the CO2 production among MSRO+ flies was 20% lower than that of MSRO- flies. Thus, mite preferences for high-metabolic-rate hosts did not explain the infection biases in this study. To assess changes in susceptibility to infection, we measured fly endurance using geotaxis assays. Older flies had lower endurance consistent with fly senescence, and this effect was magnified among MSRO+ flies. Given the biological importance of male-killing Spiroplasma, potential changes in the interactions of hosts and potential vectors could impact the ecology and evolution of host species. IMPORTANCE Male-killing endosymbionts are transmitted from mother to daughter and kill male offspring. Despite these major ecological effects, how these endosymbionts colonize new host species is not always clear. Mites are sometimes hypothesized to transfer these bacteria between hosts/host species. Here, we test if (i) mites prefer to infect flies that harbor Spiroplasma poulisoni MSRO and (ii) flies infected with MSRO are less able to resist mite infection. Our results show that flies infected with MSRO have weaker anti-mite resistance, but the mite preference/aversion for MSRO+ flies varied with fly age. Given the fitness and population impacts of male-killing Spiroplasma, changes in fly-mite interactions have implications for the ecology and evolution of these symbioses.

Trade-offs between reproduction and behavioural resistance against ectoparasite infection.

Reproduction is a key determinant of organismal fitness, but organisms almost always face the threat of parasite infection. Thus, potential trade-offs between mating and parasite resistance may have substantial impacts on the ecology and evolution of host species. Although trade-offs between microbial resistance and mating in arthropods are well-documented, there is a paucity of evidence that mating compromises host resistance to the ubiquitous threat posed by ectoparasites. Despite the centrality of reproduction to host fitness and the widespread risk of parasites, there is a dearth of experiments showing a trade-off between mating/reproduction and anti-parasite behaviours. In this study, we test if mating increases the susceptibility of female flies to mite infection. We also investigated a potential underlying mechanism for the trade-off: that mating reduces overall endurance and hence anti-parasitic defenses among female flies. We experimentally mated female Drosophila nigrospiracula, with or without a chance to recover from male harassment, and challenged them with a natural ectoparasite, the mite Macrocheles subbadius. Mated females, regardless of time for recovery from male harassment, acquired more infections than unmated females. Furthermore, mated females had lower endurance in negative geotaxis assays, suggesting the increased susceptibility is due to reduced endurance. Our research shows a trade-off between reproduction and parasite resistance in a host-macroparasite system and suggests that trade-off theory is a fruitful direction for understanding these associations.

Extending the ecology of fear: Parasite-mediated sexual selection drives host response to parasites.

The 'ecology of fear' describes the negative effects natural enemies have on potential victims even when those victims are not consumed or infected. Although recent work has demonstrated parasites have non-consumptive effects (NCE) on potential hosts, how these effects vary within host populations is not well understood. We investigated how NCE vary based on host risk of infection and relative cost of infection by measuring the metabolic rate (MR) of naive Drosophila nigrospiracula exposed to an ectoparasite, Macrocheles subbadius. We tested two mutually exclusive hypotheses: 1) asymmetrical costs of infection drive adaptions for stronger responses to parasite exposure; or 2) asymmetrical risks of infection drive adaptions for stronger responses to parasite exposure. In this system, male flies have higher costs of infection relative to female flies due to parasite-mediated sexual selection; similarly, virgin females experience higher costs of infection relative to mated females. Risk of infection also varies among flies because mites preferentially infect female flies over males, and mites preferentially infect mated females over virgin females. Our results were compatible with the hypothesis that costs of infection drive the strength of response to mite risk. Female flies responded to parasite exposure with a 15.1% increase in MR, while exposed males showed a stronger response with a 31.3% increase in MR. Mated females increased their MR by 34.8% during mite exposure whereas virgin females experienced an increase of 61.2%. Our findings suggest that NCE of parasites can vary based on state-dependent costs of infection.

Ecology of fear: environment-dependent parasite avoidance among ovipositing Drosophila.

Habitat avoidance is an anti-parasite behaviour exhibited by at-risk hosts that can minimize exposure to parasites. Because environments are often heterogeneous, host decision-making with regards to habitat use may be affected by the presence of parasites and habitat quality simultaneously. In this study we examine how the ovipositing behaviour of a cactiphilic fruit fly, Drosophila nigrospiracula, is affected by the presence of an ectoparasitic mite, Macrocheles subbadius, in conjunction with other environmental factors - specifically the presence or absence of conspecific eggs and host plant tissue. We hypothesized that the trade-off between site quality and parasite avoidance should favour ovipositing at mite-free sites even if it is of inferior quality. We found that although flies avoided mites in homogeneous environments (86% of eggs at mite-free sites), site quality overwhelmed mite avoidance. Both conspecific eggs (65% of eggs at infested sites with other Drosophila eggs) and host plant tissue (78% of eggs at infested sites with cactus) overpowered mite avoidance. Our results elucidate the context-dependent decision-making of hosts in response to the presence of parasites in variable environments, and suggest how the ecology of fear and associated trade-offs may influence the relative investment in anti-parasite behaviour in susceptible hosts.

Proximity to parasites reduces host fitness independent of infection in a Drosophila-Macrocheles system.

Parasites are known to have direct negative effects on host fitness; however, the indirect effects of parasitism on host fitness sans infection are less well understood. Hosts undergo behavioural and physiological changes when in proximity to parasites. Yet, there is little experimental evidence showing that these changes lead to long-term decreases in host fitness. We aimed to determine if parasite exposure affects host fitness independent of contact, because current approaches to parasite ecology may underestimate the effect of parasites on host populations. We assayed the longevity and reproductive output of Drosophila nigrospiracula exposed or not exposed to ectoparasitic Macrocheles subbadius. In order to preclude contact and infection, mites and flies were permanently separated with a mesh screen. Exposed flies had shorter lives and lower fecundity relative to unexposed flies. Recent work in parasite ecology has argued that parasite-host systems show similar processes as predator-prey systems. Our findings mirror the non-consumptive effects observed in predator-prey systems, in which prey species suffer reduced fitness even if they never come into direct contact with predators. Our results support the perspective that there are analogous effects in parasite-host systems, and suggest new directions for research in both parasite ecology and the ecology of fear.

Host Respiration Rate and Injury-Derived Cues Drive Host Preference by an Ectoparasite of Fruit Flies.

Host bioenergetics and energy fluxes can be applied to measure the ecological and physiological effects of parasitism. By measuring changes in host metabolic rate, one can estimate the physiological costs of infection. Additionally, metabolic rate dictates the rate of resource conversion within a host and, by extension, the resources available to a parasite. We hypothesize that parasites are selected to respond to cues that indicate high resource availability, that is, host metabolic state. We investigated whether an ectoparasite mite (Macrocheles subbadius) can differentiate between potential hosts (Drosophilia nigrospiracula) on the basis of relative carbon dioxide output as measured by respirometry. In pairwise choice tests, mites were allowed to choose between two size-matched fruit flies with differing metabolic rates or levels of CO2 output. Our results showed that mites preferentially infect flies with relatively higher respiration rates. Accordingly, we investigated whether fly respiratory rate (measured by CO2 production) changes in response to injury, potentially explaining a previously reported preference for injured flies. We also tested whether chemical cues released during injury influence preference for injured hosts. We determined that fly exudate (mostly consisting of hemolymph) applied to an uninjured fly released at the site of injury significantly increased the likelihood of infection, but injury did not significantly change the CO2 output of the flies. Our results suggest that parasites are relying on chemical cues not only for host finding but also to discriminate between hosts on the basis of the rate of respiration, with potentially important implications for the metabolic theory of ecology.

Mitey Costly: Energetic Costs of Parasite Avoidance and Infection.

Parasites reduce host fitness via perturbations to host energy allocation, growth, survival, and reproduction. Here, we investigate the independent effects of parasite exposure and infection on host metabolic rate. Our study focuses on Drosophila hydei and a naturally occurring ectoparasitic mite, Macrocheles muscaedomesticae. We use flow-through respirometry to measure the metabolic rate of flies during the period of exposure (preinfection) and during mite attachment. Flies were exposed to mites either indirectly (through a mesh screen) or directly, allowing for physical contact between the fly and the mite. We predicted that fly metabolic rate would increase with the level of parasite exposure: unexposed flies < flies with indirect exposure to mites < flies with direct contact with mites < flies actively infected with mites. As expected, flies indirectly exposed to but not in direct contact with mites produced 70% more CO2 than unexposed flies. Flies in direct contact with mites produced 35% more CO2 than flies with indirect contact, and this was more than double the amount of CO2 produced by unexposed flies. However, infected flies-those actually carrying mites-did not produce significantly more CO2 than uninfected flies. Our results show that simply being exposed to mites, either indirectly or directly, was sufficient to elicit a response from the host in terms of elevated CO2 production. Our results show that the costs of parasitism can potentially extend beyond the physiological costs of infection per se to include the energetic costs associated with parasite avoidance. Although studies have shown energetic costs associate with predator-avoidance behaviors, no study to our knowledge has measured the metabolic cost of parasite avoidance.