Proportional fitness loss and the timing of defensive investment: a cohesive framework across animals and plants.

The risk of consumption is a pervasive aspect of ecology and recent work has focused on synthesis of consumer-resource interactions (e.g., enemy-victim ecology). Despite this, theories pertaining to the timing and magnitude of defenses in animals and plants have largely developed independently. However, both animals and plants share the common dilemma of uncertainty of attack, can gather information from the environment to predict future attacks and alter their defensive investment accordingly. Here, we present a novel, unifying framework based on the way an organism's ability to defend itself during an attack can shape their pre-attack investment in defense. This framework provides a useful perspective on the nature of information use and variation in defensive investment across the sequence of attack-related events, both within and among species. It predicts that organisms with greater proportional fitness loss if attacked will gather and respond to risk information earlier in the attack sequence, while those that have lower proportional fitness loss may wait until attack is underway. This framework offers a common platform to compare and discuss consumer effects and provides novel insights into the way risk information can propagate through populations, communities, and ecosystems.

Variation in both host defense and prior herbivory can alter plant-vector-virus interactions.

BACKGROUND: While virus-vector-host interactions have been a major focus of both basic and applied ecological research, little is known about how different levels of plant defense interact with prior herbivory to affect these relationships. We used genetically-modified strains of tomato (Solanum lycopersicum) varying in the jasmonic acid (JA) plant defense pathways to explore how plant defense and prior herbivory affects a plant virus (tomato yellow leaf curl virus, 'TYLCV'), its vector (the whitefly Bemisia tabaci MED), and the host. RESULTS: Virus-free MED preferred low-JA over high-JA plants and had lower fitness on high-JA plants. Viruliferous MED preferred low-JA plants but their survival was unaffected by JA levels. While virus-free MED did not lower plant JA levels, viruliferous MED decreased both JA levels and the expression of JA-related genes. Infestation by viruliferous MED reduced plant JA levels. In preference tests, neither virus-free nor viruliferous MED discriminated among JA-varying plants previously exposed to virus-free MED. However, both virus-free and viruliferous MED preferred low-JA plant genotypes when choosing between plants that had both been previously exposed to viruliferous MED. The enhanced preference for low-JA genotypes appears linked to the volatile compound neophytadiene, which was found only in whitefly-infested plants and at concentrations inversely related to plant JA levels. CONCLUSIONS: Our findings illustrate how plant defense can interact with prior herbivory to affect both a plant virus and its whitefly vector, and confirm the induction of neophytadiene by MED. The apparent attraction of MED to neophytadiene may prove useful in pest detection and management.

Asymmetric biotic interactions and abiotic niche differences revealed by a dynamic joint species distribution model.

A species' distribution and abundance are determined by abiotic conditions and biotic interactions with other species in the community. Most species distribution models correlate the occurrence of a single species with environmental variables only, and leave out biotic interactions. To test the importance of biotic interactions on occurrence and abundance, we compared a multivariate spatiotemporal model of the joint abundance of two invasive insects that share a host plant, hemlock woolly adelgid (HWA; Adelges tsugae) and elongate hemlock scale (EHS; Fiorina externa), to independent models that do not account for dependence among co-occurring species. The joint model revealed that HWA responded more strongly to abiotic conditions than EHS. Additionally, HWA appeared to predispose stands to subsequent increase of EHS, but HWA abundance was not strongly dependent on EHS abundance. This study demonstrates how incorporating spatial and temporal dependence into a species distribution model can reveal the dependence of a species' abundance on other species in the community. Accounting for dependence among co-occurring species with a joint distribution model can also improve estimation of the abiotic niche for species affected by interspecific interactions.

Chronic impacts of invasive herbivores on a foundational forest species: a whole-tree perspective.

Forests make up a large portion of terrestrial plant biomass, and the long-lived woody plants that dominate them possess an array of traits that deter consumption by forest pests. Although often extremely effective against native consumers, invasive species that avoid or overcome these defenses can wreak havoc on trees and surrounding ecosystems. This is especially true when multiple invasive species co-occur, since interactions between invasive herbivores may yield non-additive effects on the host. While the threat posed by invasive forest pests is well known, long-term field experiments are necessary to explore these consumer-host interactions at appropriate spatial and temporal scales. Moreover, it is important to measure multiple variables to get a "whole-plant" picture of their combined impact. We report the results of a 4-yr field experiment addressing the individual and combined impacts of two invasive herbivores, the hemlock woolly adelgid (Adelges tsugae) and elongate hemlock scale (Fiorinia externa), on native eastern hemlock (Tsuga canadensis) in southern New England. In 2011, we planted 200 hemlock saplings into a temperate forest understory and experimentally manipulated the presence/absence of both herbivore species; in 2015, we harvested the 88 remaining saplings and assessed plant physiology, growth, and resource allocation. Adelgids strongly affected hemlock growth: infested saplings had lower above/belowground biomass ratios, more needle loss, and produced fewer new needles than control saplings. Hemlock scale did not alter plant biomass allocation or growth, and its co-occurrence did not alter the impact of adelgid. While both adelgid and scale impacted the concentrations of primary metabolites, adelgid effects were more pronounced. Adelgid feeding simultaneously increased free amino acids local to feeding sites and a ~30% reduction in starch. The cumulative impact of adelgid-induced needle loss, manipulation of nitrogen pools, and the loss of stored resources likely accelerates host decline through disruption of homeostatic source-sink dynamics occurring at the whole-plant level. Our research stresses the importance of considering long-term impacts to predict how plants will cope with contemporary pressures experienced in disturbed forests.

Two invasive herbivores on a shared host: patterns and consequences of phytohormone induction.

Herbivore-induced changes in host quality mediate indirect interactions between herbivores. The nature of these indirect interactions can vary depending on the identity of herbivores involved, species-specific induction of defense-signaling pathways, and sequence of attack. However, our understanding of the role of these signaling pathways in the success of multiple exotic herbivores is less known. Eastern hemlock (Tsuga canadensis) is attacked by two invasive herbivores [elongate hemlock scale (EHS; Fiorinia externa) and hemlock woolly adelgid (HWA; Adelges tsugae)] throughout much of its range, but prior attack by EHS is known to deter HWA. The potential role of phytohormones in this interaction is poorly understood. We measured endogenous levels of phytohormones in eastern hemlock in response to attack by these invasive herbivores. We also used exogenous application of methyl jasmonate (MJ) and acibenzolar-S-methyl (ASM), a salicylic acid (SA) pathway elicitor, to test the hypothesis that defense-signaling phytohormones typically induced by herbivores could deter HWA. Resistance to adelgid attack was assessed using a behavioral assay. Adelgid feeding significantly elevated both abscisic acid (ABA) and SA in local tissues, while EHS feeding had no detectable effect on either phytohormone. HWA progrediens and sistens crawlers preferred to settle on ASM-treated foliage. In contrast, HWA crawlers actively avoided settlement on MJ-treated foliage. We suggest that induction of ABA- and SA-signaling pathways, in concert with defense-signaling interference, may aid HWA invasion success, and that defense-signaling interference, induced by exotic competitors, may mediate resistance of native hosts.

Social buffering in a eusocial invertebrate: termite soldiers reduce the lethal impact of competitor cues on workers.

While the impact of predator-induced stress on prey has received considerable attention, there has been far less research into the effect of competitors. Cues from aggressive competitors should be particularly likely to evoke behavioral and/or physiological responses, since they may be indicative of both direct (interference) and indirect (exploitative) threats. The danger posed by such competitors, and the "fear" they evoke, should be reduced at lower competitor densities and by the presence of individual conspecifics specialized for defense. We assessed how Reticulitermes flavipes termite workers and soldiers were affected by cues from conspecific nestmates, conspecific non-nestmates, and the heterospecific competitor R. virginicus. Competitor cues altered flavipes worker and soldier behavior, decreasing worker growth and increasing their mortality. The presence of flavipes soldiers largely ameliorated these negative impacts: adding even a single soldier (5% of flavipes individuals) decreased worker mortality by 50-80%. Although worker mortality increased with competitor density, increased soldier densities did not increase the benefit to workers. The small number of soldiers required to substantially alter cue-mediated interactions suggests that this caste, in addition to providing direct defense, also occupies a "keystone role" by providing homeostatic feedback to workers functioning in stressful environments.

Dropping Behavior in the Pea Aphid (Hemiptera: Aphididae): How Does Environmental Context Affect Antipredator Responses?

The pea aphid Acyrthosiphon pisum : Harris (Hemiptera: Aphididae) is a phloem-feeding insect whose antipredator defenses include kicking, walking away, and dropping from the plant. Aphid dropping, a risky and energetically costly antipredator behavior, can be increased by the release of aphid alarm pheromone; there is also evidence that insect density and plant health can affect the likelihood of aphids engaging in this behavior. We investigated whether interactions between alarm cues, insect density, and plant health can alter the dropping behavior of aphids in response to an artificial disturbance. The presence of the alarm pheromone E-ß-farnesene resulted in a nearly 15-fold increase in aphid dropping behavior; the other two factors, however, did not affect dropping and none of the two- or three-way interactions were significant. This was surprising because aphids affected plant health: production of new plant biomass after 5 d of exposure to high aphid densities was 50% lower than in the control treatment. This research adds to our understanding of the factors affecting aphid antipredator behavior; the fact that neither aphid density nor feeding period impacted dropping may reflect the high energetic costs of this activity and an unwillingness to use it in any but the riskiest situations.

Insecticides promote viral outbreaks by altering herbivore competition.

While the management of biological invasions is often characterized by a series of single-specieg decisions, invasive species exist within larger food webs. These biotic interactions can alter the impact of control/eradication programs and may cause suppression efforts to inadvertently facilitate invasion spread and impact. We document the rapid replacement of the invasive Bemisia Middle East-Asia Minor I (MEAM1) cryptic biotype by the cryptic Mediterranean (MED) biotype throughout China and demonstrate that MED is more tolerant of insecticides and a better vector of tomato yellow leaf curl virus (TYLCV) than MEAMJ. While MEAM1 usually excludes MED under natural conditions, insecticide application reverses the MEAM1-MED competitive hierarchy and allows MED to exclude MEAMI. The insecticide-mediated success of MED has led to TYLCV outbreaks throughout China. Our work strongly supports the hypothesis that insecticide use in China reverses the MEAMl-MED competitive hierarchy and allows MED to displace MEAM1 in managed landscapes. By promoting the dominance of a Bemisia species that is a competent viral vector, insecticides thus increase the spread and impact of TYLCV in heterogeneous agroecosystems.

Manipulation of Host Quality and Defense by a Plant Virus Improves Performance of Whitefly Vectors.

Pathogen-mediated interactions between insect vectors and their host plants can affect herbivore fitness and the epidemiology of plant diseases. While the role of plant quality and defense in mediating these tripartite interactions has been recognized, there are many ecologically and economically important cases where the nature of the interaction has yet to be characterized. The Bemisia tabaci (Gennadius) cryptic species Mediterranean (MED) is an important vector of tomato yellow leaf curl virus (TYLCV), and performs better on virus-infected tomato than on uninfected controls. We assessed the impact of TYLCV infection on plant quality and defense, and the direct impact of TYLCV infection on MED feeding. We found that although TYLCV infection has a minimal direct impact on MED, the virus alters the nutritional content of leaf tissue and phloem sap in a manner beneficial to MED. TYLCV infection also suppresses herbivore-induced production of plant defensive enzymes and callose deposition. The strongly positive net effect on TYLCV on MED is consistent with previously reported patterns of whitefly behavior and performance, and provides a foundation for further exploration of the molecular mechanisms responsible for these effects and the evolutionary processes that shape them.

Multiple forms of vector manipulation by a plant-infecting virus: Bemisia tabaci and tomato yellow leaf curl virus.

For many insect-vectored plant viruses, the relationship between feeding behavior and vector competence may prove integral to an understanding of the epidemiology of the resulting plant disease. While plant-infecting viruses are well known to change host plant physiology in a way that makes them more attractive to vectors, viral manipulation of the vectors themselves has only recently been reported. Previous research suggested that the rapid spread of Tomato yellow leaf curl virus (TYLCV) throughout China has been facilitated by its primary vector, the whitefly Bemisia tabaci. We conducted two experiments testing the impact of TYLCV infection of the host plant (tomato) and vector (B. tabaci biotypes B and Q) on whitefly feeding behavior. Whiteflies of biotypes B and Q both appeared to find TYLCV-infected plants more attractive, probing them more quickly and having a greater number of feeding bouts; this did not, however, alter the total time spent feeding. Viruliferous whiteflies fed more readily than uninfected whiteflies and spent more time salivating into sieve tube elements. Because vector salivation is essential for viral transmission, this virally mediated alteration of behavior should provide TYLCV a direct fitness benefit. This is the first report of such manipulation by a nonpropagative virus that belongs to an exclusively plant-infecting family of viruses (Geminiviridae). In the context of previous research showing that feeding on TYLCV-infected plants harms biotype B but helps biotype Q, the fact that both biotypes were equally affected by TYLCV also suggests that the virus may alter the biotype B-biotype Q competitive interaction in favor of biotype Q.

Are exotic herbivores better competitors? A meta-analysis.

Competition plays an important role in structuring the community dynamics of phytophagous insects. As the number and impact of biological invasions increase, it has become increasingly important to determine whether competitive differences exist between native and exotic insects. We conducted a meta-analysis to test the hypothesis that native/ exotic status affects the outcome of herbivore competition. Specifically, we used data from 160 published studies to assess plant-mediated competition in phytophagous insects. For each pair of competing herbivores, we determined the native range and coevolutionary history of each herbivore and host plant. Plant-mediated competition occurred frequently, but neither native nor exotic insects were consistently better competitors. Spatial separation reduced competition in native insects but showed little effect on exotics. Temporal separation negatively impacted native insects but did not affect competition in exotics. Insects that coevolved with their host plant were more affected by interspecific competition than herbivores that lacked a coevolutionary history. Insects that have not coevolved with their host plant may be at a competitive advantage if they overcome plant defenses. As native/exotic status does not consistently predict outcomes of competitive interactions, plant-insect coevolutionary history should be considered in studies of competition.

Failure under stress: the effect of the exotic herbivore Adelges tsugae on biomechanics of Tsuga canadensis.

BACKGROUND AND AIMS: Exotic herbivores that lack a coevolutionary history with their host plants can benefit from poorly adapted host defences, potentially leading to rapid population growth of the herbivore and severe damage to its plant hosts. The hemlock woolly adelgid (Adelges tsugae) is an exotic hemipteran that feeds on the long-lived conifer eastern hemlock (Tsuga canadensis), causing rapid mortality of infested trees. While the mechanism of this mortality is unknown, evidence indicates that A. tsugae feeding causes a hypersensitive response and alters wood anatomy. This study investigated the effect of A. tsugae feeding on biomechanical properties at different spatial scales: needles, twigs and branches. METHODS: Uninfested and A. tsugae-infested samples were collected from a common garden experiment as well as from naturally infested urban and rural field sites. Tension and flexure mechanical tests were used to quantify biomechanical properties of the different tissues. In tissues that showed a significant effect of herbivory, the potential contributions of lignin and tissue density on the results were quantified. KEY RESULTS: Adelges tsugae infestation decreased the abscission strength, but not flexibility, of needles. A. tsugae feeding also decreased mechanical strength and flexibility in currently attacked twigs, but this effect disappeared in older, previously attacked branches. Lignin and twig tissue density contributed to differences in mechanical strength but were not affected by insect treatment. CONCLUSIONS: Decreased strength and flexibility in twigs, along with decreased needle strength, suggest that infested trees experience resource stress. Altered growth patterns and cell wall chemistry probably contribute to these mechanical effects. Consistent site effects emphasize the role of environmental variation in mechanical traits. The mechanical changes measured here may increase susceptibility to abiotic physical stressors in hemlocks colonized by A. tsugae. Thus, the interaction between herbivore and physical stresses is probably accelerating the decline of eastern hemlock, as HWA continues to expand its range.

Terpene chemistry of eastern hemlocks resistant to hemlock woolly adelgid.

Recent studies have identified a small number of individual eastern hemlock trees that demonstrate relative resistance to the introduced sap-feeding insect, the hemlock woolly adelgid. Using gas chromatography, we compared concentrations of twenty-two terpenoids in susceptible and relatively resistant trees, both in the forest and in propagated cuttings in a common-garden setting. Terpenoid concentrations were higher in twig tissue of resistant versus susceptible trees, across six sampling dates and at both sites. These changes may be particularly important because the hemlock woolly adelgid feeds on twig tissue. Because the common-garden cuttings were free of herbivores, the higher terpenoid concentrations are interpreted as a constitutive defense. Increased levels of monoterpenes and sesquiterpenes imply an overall increase in the input of carbon precursors to both terpenoid synthesis pathways.

Rapamycin attenuates reconsolidation of a backwards-conditioned aversive stimuli in female mice.

RATIONALE: The mammalian target of rapamycin (mTOR) kinase is known to mediate consolidation and reconsolidation of aversive memories. Most studies in this area use a forward conditioning paradigm in which the conditioned stimulus (CS) precedes the unconditioned stimulus (US). Little is known, however, about the neurobiological underpinnings of backwards (BW) conditioning paradigms, particularly in female mice. In BW conditioning, the CS does not become directly associated with the US; it instead evokes conditioned fear by reactivating a memory of the conditioning context and indirectly retrieving a memory of the aversive US. OBJECTIVES: We sought to examine BW conditioned fear memory processes in female mice. First, we examined whether freezing to a BW CS is mediated by fear to the conditioning context. Second, we tested whether blocking consolidation of a BW CS attenuated memory of the CS and conditioning context. Finally, we tested whether blocking reconsolidation of a BW CS attenuated memory of the conditioning context. RESULTS: We show that conditioned freezing to a BW CS is mediated by fear to the conditioning context. Furthermore, rapamycin-an mTOR inhibitor, when given immediately following BW conditioning, impairs consolidation of both cued and contextual fear memory. Similarly, rapamycin given following retrieval of a BW CS blocks context recall. Rapamycin is acting on reconsolidation as CS retrieval is necessary to see the effects of rapamycin on context memory recall. CONCLUSIONS: Our study provides novel evidence that indirect retrieval cues are sensitive to rapamycin in female mice. The capacity to indirectly reactivate memories and render them susceptible to disruption is critical in the translation of reconsolidation-based approaches to the clinic.

Domestication reduces caterpillar response to auditory predator cues.

Domestication can lead to significant changes in the growth and behavior of organisms. While the threat of predation is a strong selective force in the wild, the relaxation or removal of this threat in captive-rearing environments selects for reduced sensitivity to biotic stressors. Previous work has documented such changes in other taxa, but no work has been done on domestication-related losses of predation risk sensitivity in insects. We exposed both wild and domesticated (>50 generations in captivity) Lymantria dispar dispar (Lepidoptera: Erebidae) larvae to recordings of predators (wasp buzzing), nonpredators (mosquito buzzing), or no sound to compare the effects of predation risk on the two stocks. Wasp buzzing, but not mosquito buzzing, decreased survival of wild caterpillars relative to the control; domesticated caterpillars showed no such response. Domesticated L. dispar larvae appear to have reduced sensitivity to predation risk cues, suggesting that captive-reared insects may not always be analogs to their wild counterparts for risk-related behavioral studies.

The cost of safety: refuges increase the impact of predation risk in aquatic systems.

Although use of refuge habitats by prey can reduce their risk of predation, refuge use may also involve costs such as increased within-refuge competition for resources. Despite the ubiquity of refuge use by prey, it is unknown whether predator-induced use of refuges has widespread, negative nonconsumptive effects on prey growth, survival, and fecundity. We performed a meta-analysis of 204 studies of aquatic taxa containing data on 271 distinct predator--prey pairs and found strong evidence that the negative effect of predation risk on prey activity, growth, and fecundity increases when prey have access to refuge habitats. Moreover, the effect of refuge habitats on growth and activity depends upon whether the refuge provides partial or total protection from predators. These results suggest that prey choosing whether to use refuges face a trade-off between lowering the immediate risk of being consumed and increased nonconsumptive costs of refuge use. Our results suggest that changes in nonconsumptive effects in the presence of refuge habitats may alter prey population dynamics, coexistence, and metapopulation dynamics. Moreover, our results reveal key pragmatic considerations: the magnitude and direction of nonconsumptive effects may depend on the presence of refuge habitat and whether the refuge provides partial or total protection from predators.

Auditory predator cues decrease herbivore survival and plant damage.

The high fitness cost of predation selects prey capable of detecting risk cues and responding in ways that reduce their vulnerability. While the impacts of auditory predator cues have been extensively researched in vertebrate prey, much less is known about invertebrate species' responses and their potential to affect the wider food web. We exposed larvae of Spodoptera exigua, a slow-moving and vulnerable herbivore hunted by aerial predators, to recordings of wasp buzzing (risk cue), mosquito buzzing (no-risk cue), or a no-sound control in both laboratory and field settings. In the laboratory, wasp buzzing (but not mosquito buzzing) reduced survival relative to the control; there was, however, no effect on time to or weight at pupation in survivors. In the field, wasp buzzing reduced caterpillar herbivory and increased plant biomass relative to the control treatment. In contrast, mosquito buzzing reduced herbivory less than wasp buzzing and had no effect on plant biomass. The fact that wasp cues evoked strong responses in both experiments, while mosquito buzzing generally did not, indicates that caterpillars were responding to predation risk rather than sound per se. Such auditory cues may have an important but largely unappreciated impacts on terrestrial invertebrate herbivores and their host plants.

A short pre-conception bout of predation risk affects both children and grandchildren.

Traumatic events that affect physiology and behavior in the current generation may also impact future generations. We demonstrate that an ecologically realistic degree of predation risk prior to conception causes lasting changes in the first filial (F1) and second filial (F2) generations. We exposed male and female mice to a live rat (predator stress) or control (non-predator) condition for 5 min. Ten days later, stressed males and females were bred together as were control males and females. Adult F1 offspring from preconception-stressed parents responded to a mild stressor with more anxiety-like behavior and hyperarousal than offspring from control parents. Exposing these F1 offspring to the mild stressor increased neuronal activity (cFOS) in the hippocampus and altered glucocorticoid system function peripherally (plasma corticosterone levels). Even without the mild stressor, F1 offspring from preconception-stressed parents still exhibited more anxiety-like behaviors than controls. Cross-fostering studies confirmed that preconception stress, not maternal social environment, determined offspring behavioral phenotype. The effects of preconception parental stress were also unexpectedly persistent and produced similar behavioral phenotypes in the F2 offspring. Our data illustrate that a surprisingly small amount of preconception predator stress alters the brain, physiology, and behavior of future generations. A better understanding of the 'long shadow' cast by fearful events is critical for understanding the adaptive costs and benefits of transgenerational plasticity. It also suggests the intriguing possibility that similar risk-induced changes are the rule rather than the exception in free-living organisms, and that such multigenerational impacts are as ubiquitous as they are cryptic.

Modeling range dynamics in heterogeneous landscapes: invasion of the hemlock woolly adelgid in eastern North America.

Range expansion by native and exotic species will continue to be a major component of global change. Anticipating the potential effects of changes in species distributions requires models capable of forecasting population spread across realistic, heterogeneous landscapes and subject to spatiotemporal variability in habitat suitability. Several decades of theory and model development, as well as increased computing power and availability of fine-resolution GIS data, now make such models possible. Still unanswered, however, is the question of how well this new generation of dynamic models will anticipate range expansion. Here we develop a spatially explicit stochastic model that combines dynamic dispersal and population processes with fine-resolution maps characterizing spatiotemporal heterogeneity in climate and habitat to model range expansion of the hemlock woolly adelgid (HWA; Adelges tsugae). We parameterize this model using multiyear data sets describing population and dispersal dynamics of HWA and apply it to eastern North America over a 57-year period (1951-2008). To evaluate the model, the observed pattern of spread of HWA during this same period was compared to model predictions. Our model predicts considerable heterogeneity in the risk of HWA invasion across space and through time, and it suggests that spatiotemporal variation in winter temperature, rather than hemlock abundance, exerts a primary control on the spread of HWA. Although the simulations generally matched the observed current extent of the invasion of HWA and patterns of anisotropic spread, it did not correctly predict when HWA was observed to arrive in different geographic regions. We attribute differences between the modeled and observed dynamics to an inability to capture the timing and direction of long-distance dispersal events that substantially affected the ensuing pattern of spread.

Exotic herbivores on a shared native host: tissue quality after individual, simultaneous, and sequential attack.

Plants in nature are often attacked by multiple enemies whose effect on the plant cannot always be predicted based on the outcome of individual attacks. We investigated how two invasive herbivores, the hemlock woolly adelgid (Adelges tsugae) (HWA) and the elongate hemlock scale (Fiorinia externa) (EHS), alter host plant quality (measured as amino acid concentration and composition) when feeding individually or jointly on eastern hemlock (Tsuga canadensis), an important long-lived forest tree that is in severe decline. The joint herbivore treatments included both simultaneous and sequential infestations by the two herbivores. We expected resource depletion over time, particularly in response to feeding by HWA. In contrast, HWA dramatically increased the concentration and altered the composition of individual free amino acids. Compared to control trees, HWA increased total amino acid concentration by 330% after 1 year of infestation. Conversely, EHS had a negligible effect when feeding individually. Interestingly, there was a marginally significant HWA × EHS interaction that suggests the potential for EHS presence to reduce the impact of HWA on foliage quality when the two species co-occur. We suggest indirect effects of water stress as a possible physiological mechanism for our results. Understanding how species interactions change the physiology of a shared host is crucial to making more accurate predictions about host mortality and subsequent changes in affected communities and ecosystems, and to help design appropriate management plans.