Assessing the risk effects of native predators on the exotic American bullfrog (Lithobates catesbeianus) and their indirect consequences to ecosystem function

Understanding the factors and mechanisms that affect the impacts of invasive species in invaded environments has been widely debated among researchers. However, few studies about invasive species have explored the effects of predation risks by native predators on exotic prey. Herein, the traditional invasive predator-native prey framework was reversed. We tested if tadpoles, of the worldwide invasive American Bullfrog Lithobates catesbeianus, were affected by the predation risk imposed by native predators. We used two different species of belostomatid predators and tested whether and how predation-induced phenotypic plasticity on L. catesbeianus reverberated in morphological, physiological, and ecosystem-level processes. Individuals of L. catesbeianus modified their morphological (tail muscle width), behavioral (activity and foraging), and physiological (growth and growth efficiency) traits in the presence of predation risk. Based on the observed morphological changes, our results suggest that prey may recognize predator-specific cues. In addition, we observed that L. catesbeianus' responses to predation risk can affect ecosystem-level properties, by inducing trophic cascades and reducing animal-mediated nutrient recycling rates. In summary, our study supports that exotic prey species who are subjected to native predators may display anti-predator responses, with implications for their development, as well as possible ecosystem-level effects.     

Consequences of an amphibian malformity for development and fitness in complex environments

1. Environmental stressors have both lethal and sublethal effects, such as altered developmental rates and the induction of malformations. Ecological interactions, including predation and competition, often amplify such effects, for instance by inducing behavioural changes that increase susceptibility to the stress. 2. Using experimental mesocosms, we asked whether the density of conspecific competitors and predation risk from larval water beetles (Dytiscus spp.) affect the development of malformations in tadpoles of the wood frog (Rana sylvatica). We also examined whether such malformities increase the susceptibility of tadpoles to predation. 3. The risk of predation decreased the frequency of malformities in both low‐ and high‐density treatments, although this effect was greater at low density. Behavioural observations suggested that reductions in activity by amphibian larvae induced by predators mediated these responses by decreasing cumulative exposure to ultraviolet‐B radiation, the putative stressor causing the observed malformity. These results suggest that predators can reduce negative impacts of stressors by inducing behavioural changes in prey organisms. 4. Malformed individuals were twice as vulnerable to predators as non‐malformed individuals, suggesting that sublethal effects can ultimately cause increased mortality.     

Prey risk assessment depends on conspecific density

In many systems, the number of prey killed by predators increases with prey density. This in turn generates higher levels of the indirect signals that prey use to assess predation risk. A model developed by Peacor (2003) showed that prey that respond to predator cues without accounting for conspecific density will consistently over‐ or under‐estimate risk and therefore invest improperly in anti‐predator defense. We tested this model using Rana temporaria tadpoles as prey and Aeshna cyanea dragonfly larvae as predators. As assumed by the model, prey reduced risky activity with increasing concentrations of predator kairomones and increased activity at high prey density. However, prey did not react to changes in cue or density if the ratio of cue‐to‐density remained constant. Prey therefore monitored their per capita risk, strongly supporting Peacor's model.     

Tadpoles of the bronze frog (Rana temporalis) assess predation risk before evoking antipredator defense behavior

Predation threat-associated behavioral response was studied in Rana temporalis tadpoles to discover the importance of predators’ visual and chemical cues (kairomones and diet-derived metabolites of consumed prey) in evoking antipredator behavior. The caged predators (dragonfly larvae) fed on prey tadpoles or insects (Notonecta spp.) and water conditioned with the predators provided the threat stimuli to the tadpole prey. The predators’ visual cues were ineffective in evoking antipredator behaviors in the tadpole prey. However, exposure to caged tadpole-fed predators or water conditioned with tadpole-fed predators elicited predator avoidance behavior in the tadpoles; they stayed away from the predators, significantly reduced swimming activity (swimming time and distance traveled), and increased burst speed. Interestingly, exposure to water conditioned with starved predators did not elicit any antipredator behavior in the prey. Further, the antipredator responses of predator-experienced tadpoles were significantly greater than those exhibited by predator-na?ve tadpoles. The study shows that R. temporalis tadpoles assess predation threat based exclusively on chemical cues emanating from the predators’ dietary metabolites and that the inclusion of conspecific prey items in the diet of the predators is perceived as a threat. The study also shows that antipredator behavior in these tadpoles is innate and is enhanced during subsequent encounters with the predators.     

Body size and group size of Cuban tree frog (<Emphasis Type="Italic">Osteopilus septentrionalis</Emphasis>) tadpoles influence their escape behaviour

Tadpoles risk attack from both aquatic and aerial predators. We investigated how body size and group size influenced the behaviour of tadpoles before and during a predatory attack from above to test the predictions of the theoretical economic escape model. We examined escape (swimming) response of small and large Cuban tree frog (Osteopilus septentrionalis) tadpoles kept under three density treatments and predicted that increased group size, body size and depth in the water column would all reduce perceived risk and, therefore, escape responses to simulated predation. Compared with the lower density groups, tadpoles in higher density groups moved shorter distances, and many individuals did not even move away in response to being touched. Contrary to our predictions based on the economic escape model, smaller tadpoles (which should be more vulnerable to a greater suite of predators) were less reactive than larger tadpoles, and this result may reflect different costs of escape. Finally, although tadpoles might be exposed to a wider range of predator species (aerial as well as aquatic predators), we found no effect of initial depth on escape responses. In conclusion, it appears that the main benefit of increased group density in O. septentrionalis tadpoles is likely to be predator dilution, and that variation in densities of tadpoles influences the escape behaviour of individual tadpoles, regardless of tadpole size.     

The advantage of no defense: predation enhances cohort survival in a desert amphibian

The impacts that predators have on prey behavior, growth, survival, and ultimately the composition of many ecological communities are mediated by prey defenses and the susceptibility of prey to predators. We hypothesized that prey populations inhabiting short-lived, species-poor, aquatic environments should lack significant morphological, developmental, and behavioral responses to predators and are therefore highly susceptible to predation. Furthermore, we predicted that the resultant decrease in prey density and increase in per capita resources due to high susceptibility to predators should enhance overall cohort survival because of enhanced growth of surviving prey. To test these ideas, we performed laboratory and outdoor mesocosm experiments to disentangle multiple effects of predators on an anuran (Scaphiopus couchii); a species highly adapted to breeding in ephemeral habitats and that has one of the shortest larval periods of all anurans. Chemical (presence of predator) and lethal predator cues (predator plus consumed conspecific) elicited no response in behavior, development, or morphology, indicating a lack of defensive mechanisms. Survivorship was significantly reduced in treatments where tadpoles were exposed to predators. However, this reduction in prey density led to accelerated time to metamorphosis, conferring an advantage to survivors who must metamorphose before ephemeral ponds dry. Our experiments demonstrated that in short-lived environments, prey may exhibit little or no response to the presence of predators presumably because selection for anti-predator defenses is countered by selection for rapid metamorphosis. However, predation actually resulted in an increase in overall cohort survival. Although predators are relatively rare in highly ephemeral aquatic environments, they may play an important role in facilitating the long-term persistence of their prey by reducing prey density.     

Chemical cues from multiple predator-prey interactions induce changes in behavior and growth of anuran larvae

Chemical signals are used as information by prey to assess predation risk in their environment. To evaluate the effects of multiple predators on prey growth, mediated by a change in prey activity, I exposed small and large bullfrog (Rana catesbeiana) larvae (tadpoles) to chemical cues from different combinations of bluegill sunfish (Lepomis macrochirus) and larval dragonfly (Anax junius) predators. Water was regularly transferred from predation trials (outdoor experiment) to aquaria (indoor experiment) in which activity and growth of tadpoles was measured. The highest predation mortality of small bullfrog larvae in the outdoor experiment was due to Anax, and it was slightly lower in the presence of both predators, probably resulting from interactions between predators. There was almost no mortality of prey with bluegill. The activity and growth of small bullfrog larvae was highest in the absence of predators and lowest in the presence of Anax. In the presence of bluegill only, or with both predators, the activity and growth of small bullfrog tadpoles was intermediate. Predators did not affect large tadpole activity and growth. Regressing mortality of small bullfrog tadpoles against activity and growth of bullfrog tadpoles revealed a significant effect for small bullfrog larvae but a non-significant effect for large bullfrog larvae. This shows that the response of bullfrog tadpoles to predators is related to their own body size. The experiment demonstrates that chemical cues are released both as predator odor and as alarm substances and both have the potential to strongly alter the activity and growth of prey. Different mechanisms by which chemical cues may be transmitted to species interactions in the food web are discussed. Received: 28 June 1999 / Accepted: 15 November 1999     

Effect of Predator-Prey Phylogenetic Similarity on the Fitness Consequences of Predation: A Trade-off between Nutrition and Disease?

A largely neglected aspect of foraging behavior is whether the costs and benefits of predation vary as a function of phylogenetic (i.e., genetic) similarity between predator and prey. Prey of varying phylogenetic similarities to predators might differ in value because both the risk of pathogen transmission and the nutritional quality of prey typically decline with decreasing phylogenetic similarity between predator and prey. I experimentally evaluated this hypothesis by feeding omnivorous spadefoot toad tadpoles (Spea bombifrons, Spea multiplicata, and Scaphiopus couchii) either conspecific tadpoles or an equal mass of three different species of heterospecific prey, all of which contained naturally occurring bacteria. I also examined which prey species Spea tadpoles preferred. I found that all three species of tadpoles performed best on, and preferred to eat, prey that were of intermediate phylogenetic similarity to the predators. Prey of intermediate phylogenetic similarity may provide the greatest fitness benefits to predators because such prey balance the nutritional benefits of closely related prey with the cost of parasite transmission between closely related individuals.     

Chemical defense of toad tadpoles under risk by four predator species

Many organisms use inducible defenses as protection against predators. In animals, inducible defenses may manifest as changes in behavior, morphology, physiology, or life history, and prey species can adjust their defensive responses based on the dangerousness of predators. Analogously, prey may also change the composition and quantity of defensive chemicals when they coexist with different predators, but such predator‐induced plasticity in chemical defenses remains elusive in vertebrates. In this study, we investigated whether tadpoles of the common toad (Bufo bufo) adjust their chemical defenses to predation risk in general and specifically to the presence of different predator species; furthermore, we assessed the adaptive value of the induced defense. We reared tadpoles in the presence or absence of one of four caged predator species in a mesocosm experiment, analyzed the composition and quantity of their bufadienolide toxins, and exposed them to free‐ranging predators. We found that toad tadpoles did not respond to predation risk by upregulating their bufadienolide synthesis. Fishes and newts consumed only a small percentage of toad tadpoles, suggesting that bufadienolides provided protection against vertebrate predators, irrespective of the rearing environment. Backswimmers consumed toad tadpoles regardless of treatment. Dragonfly larvae were the most voracious predators and consumed more predator‐naïve toad tadpoles than tadpoles raised in the presence of dragonfly cues. These results suggest that tadpoles in our experiment had high enough toxin levels for an effective defense against vertebrate predators even in the absence of predator cues. The lack of predator‐induced phenotypic plasticity in bufadienolide synthesis may be due to local adaptation for constantly high chemical defense against fishes in the study population and/or due to the high density of conspecifics.     

Cooccurrence of prey species alters the impact of predators on prey performance through multiple mechanisms

When prey are differentially affected by intra and interspecific competition, the cooccurrence of multiple prey species alters the per capita availability of food for a particular prey species which could alter how prey respond to the threat of predation, and hence the overall‐effect of predators. We conducted an experiment to examine the extent to which the nonconsumptive and overall effect of predatory water bugs on snail and tadpole traits (performance and morphology) depended on whether tadpoles and snails cooccurred. Tadpoles and snails differed in their relative susceptibility to intraspecific and interspecific competition, and predators affected both prey species via consumptive and nonconsumptive mechanisms. Furthermore, the overall effect of predators often depended on whether another prey species was present. The reasoning for why the overall effect of predators depended on whether prey species cooccurred, however, differed for each of the response variables. Predators affected snail body growth via nonconsumptive mechanisms, but the change in the overall effect of predators on snail body growth was attributable to how snails responded to competition in the absence of predators, rather than a change in how snails responded to the threat of predation. Predators did not affect tadpole body growth via nonconsumptive mechanisms, but the greater vulnerability of competitively superior prey (snails) to predators increased the strength of consumptive mechanisms (and hence the overall effect) through which predators affected tadpole growth. Predators affected tadpole morphology via nonconsumptive mechanisms, but the greater propensity for predators to kill competitively superior prey (snails) enhanced the ability of tadpoles to alter their morphology in response to the threat of predation by creating an environment where tadpoles had a higher per capita supply of food available to invest in the development of morphological defenses. Our work indicates that the mechanisms through which predators affect prey depends on the other members of the community.     

Behavioural response of bullfrog tadpoles to chemical cues of predation risk are affected by cue age and water source

When confronted by signals of predators presence, many aquatic organisms modify their phenotype (e.g., behaviour or morphology) to reduce their risk of predation. A principal means by which organisms assess predation risk is through chemical cues produced by the predators and/or prey during predation events. Such responses to predation risk can directly affect prey fitness and indirectly affect the fitness of species with which the prey interacts. Accurate assessment of the cue will affect the adaptive nature, and hence evolution, of the phenotypic response. It is therefore, important to understand factors affecting the assessment of chemical cues. Here I examined the effect of the age of chemical cues arising from an invertebrate predator, a larval dragonfly (Anax junius), which was fed bullfrog tadpoles, on the behavioural response (activity level and position) of bullfrog tadpoles. The bullfrog response to chemical cues declined as a function of chemical cue age, indicating the degradation of the chemical cue was on the order of 2–4 days. Further, the decay occurred more rapidly when the chemical cue was placed in pond water rather than well water. These results indicate a limitation of the tadpoles to interpret factors that affect the magnitude of the chemical cue and hence accurately assess predation risk. These findings also have implications for experimental design and the adaptation of phenotypic responses to chemical cues of predation risk.     

The effect of prior experience on a prey’s current perceived risk

The prior experience of prey may influence how they assess the level of predation risk associated with an information source. Here, I present the results from a set of experiments that demonstrate how the prior experience of green frog (Rana clamitans) tadpoles can influence their risk assessment during exposure to the chemical cue of predatory larval dragonflies (Anax spp.) consuming conspecific tadpoles. At the short-term scale, green frog tadpoles perceived a higher level of risk when consecutive cue exposures overlapped, but only when the total chemical cue concentration was weak. Weaker chemical cue concentrations may be less reliable than stronger cue concentrations, and overlapping cue exposures may increase the degree of certainty that tadpoles have in their perceived risk. When consecutive cue exposures did not overlap, tadpoles assessed the risk associated with each cue exposure independently. Predator-conditioned tadpoles responded longer during exposure to the Anax chemical cue than nonconditioned tadpoles, which suggests that a tadpole’s long-term experience eventually does influence its risk assessment. In general, the results suggest that a prey’s prior experience may influence its current perceived risk by influencing either the degree of certainty in or the level of its perceived risk. Understanding how the prior experience of prey influences their current risk assessment requires that the rate of decay of the value of prior experience should be identified at two timescales as an indicator of the current level of predation risk.     

Density is more important than predation risk for predicting growth and developmental outcomes in tadpoles of spotted tree frog,Litoria spenceri (Dubois 1984)

The diverse benefits of group living include protection against predators through dilution effects and greater group vigilance. However, intraspecific aggregation can decrease developmental rates and survival in prey species. We investigated the impact on tadpole development and behaviour of the interaction between population density and predation risk. Spotted tree frog (Litoria spenceri: Hylidae, Dubois 1984) tadpoles were kept at one of three different densities (two tadpoles per litre, five tadpoles per litre or 10 tadpoles per litre) until metamorphosis in the presence or absence of predatory cues. We aimed to determine the influence of population density, predation and the interaction of both factors in determining growth rates in tadpoles. Tadpoles were measured weekly to assess growth and development and filmed to quantify differences in activity and feeding frequency between groups. Generally, tadpoles housed without predators had longer developmental periods when housed with a predator, but there was no effect on tail length or total length. There was no effect of either predation cues or density on percentage of individuals feeding or moving. Although the effects of the presence of predators alone may appear to be less than the effects of the presence of competitors, the prioritisation of competitiveness over predator avoidance may increase vulnerability of tadpoles to the lethal threat of predators. This is particularly important in species such as L. spenceri, which is at risk from introduced fish predators.     

Conspecific alarm cues, but not predator cues alone, determine antipredator behavior of larval southern marbled newts, Triturus pygmaeus

Predation imposes selection on the ability of prey to recognize and respond to potential threats. Many prey species detect predators via chemoreception, particularly in aquatic environments. Also, chemical cues from injured prey are often perceived as an indication of predation risk. However, because antipredatory behavior can be costly, prey responses should depend on the current level of risk that each predator poses, which may depend on the type of chemical cues detected. We exposed larval newts, Triturus pygmaeus, to chemical cues from predator larval beetles or to alarm cues from conspecific larval newts and examined the behavioral changes of larval newts. Results showed that larval newts reduced activity levels when conspecific alarm cues were present but not when the predator cues alone were present. These results might suggest that larval newts are unable to recognize predator chemicals. To avoid costs of unnecessary antipredatory behaviors, larval newts may benefit by avoiding only predators that represent a current high level of threat, showing only antipredatory responses when they detect conspecific alarm cues indicating that an actual predatory attack has occurred.     

Predator cannibalism can shift prey community composition toward dominance by small prey species

Cannibalism among predators is a key intraspecific interaction affecting their density and foraging behavior, eventually modifying the strength of predation on heterospecific prey. Interestingly, previous studies showed that cannibalism among predators can increase or reduce predation on heterospecific prey; however, we know less about the factors that lead to these outcomes. Using a simple pond community consisting of Hynobius retardatus salamander larvae and their associated prey, I report empirical evidence that cannibalism among predators can increase predation on large heterospecific prey but reduce that on small heterospecific prey. In a field‐enclosure experiment in which I manipulated the occurrence of salamander cannibalism, I found that salamander cannibalism increased predation on frog tadpoles but reduced that on aquatic insects simultaneously. The contrasting effects are most likely to be explained by prey body size. In the study system, frog tadpoles were too large for non‐cannibal salamanders to consume, while aquatic insects were within the non‐cannibals’ consumable prey size range. However, when cannibalism occurred, a few individuals that succeeded in cannibalizing reached large enough size to consume frog tadpoles. Consequently, although cannibalism among salamanders reduced their density, salamander cannibalism increased predation on large prey frog tadpoles. Meanwhile, salamander cannibalism reduced predation on small prey aquatic insects probably because of a density reduction of non‐cannibals primarily consuming aquatic insects. Body size is often correlated with various ecological traits, for instance, diet width, consumption, and excretion rates, and is thus considered a good indicator of species’ effects on ecosystem function. All this considered, cannibalism among predators could eventually affect ecosystem function by shifting the size composition of the prey community.     

Risk of predation and behavioural response in three anuran species: influence of tadpole size and predator type

Many species alter their activity, microhabitat use, morphology and life history in response to predators. Predation risk is related to predator size and palatability of prey among others factors. We analyzed the predation risk of three species of tadpoles that occur in norwestern Patagonia, Argentina: Pleurodema thaul, Pleurodema bufoninum and Rhinella spinulosa. We sampled aquatic insect predators in 18 ponds to determine predator–tadpole assemblage in the study area. In laboratory conditions, we analysed the predation rate imposed by each predator on each tadpole species at different tadpole sizes. Finally, we tested whether tadpoles alter their activity in the presence of chemical and visual cues from predators. Small P. thaul and P. bufoninum tadpoles were the most vulnerable prey species, while small R. spinulosa tadpoles were only consumed by water bugs. Dragonflies and water bugs were the most dangerous tadpole predators. Small P. thaul tadpoles reduced their activity when they were exposed to all predators, while large tadpoles only reduced the activity in the presence of large predators (dragonfly larvae and water bugs). Small P. bufoninum tadpoles reduced the activity when they were exposed to beetle larvae and dragonfly larvae, while large tadpoles only reduced activity when they were exposed to larger predators (water bugs and dragonfly larvae). R. spinulosa tadpoles were the less sensitive to presence of predators, only larger tadpoles responded significantly to dragonfly larvae by reducing their activity. We conclude that behavioural responses of these anuran species were predator-specific and related to the risk imposed by each predator.     

Antipredator Behavior of American Bullfrogs (Lithobates catesbeianus) in a Novel Environment

Invasive species capable of recognizing potential predators may have increased establishment rates in novel environments. Individuals may retain historical predator recognition and invoke innate responses in the presence of taxonomically or ecologically similar predators, generalize antipredator responses, or learn to avoid risky species in novel environments. Invasive amphibians in aquatic environments often use chemical cues to assess predation risk and learn to avoid novel predators via direct experience and/or associated chemical cues. Ontogeny may also influence recognition; experience with predators may need to occur at certain developmental stages for individuals to respond correctly. We tested predator recognition in invasive American bullfrog ( Lithobates catesbeianus) tadpoles that varied in experience with fish predators at the population and individual scale. We found that bullfrog tadpoles responded to a historical predator, largemouth bass ( Micropterus salmoides), only if the population was locally sympatric with largemouth bass. Individuals from a population that did not co‐occur with largemouth bass did not increase refuge use in response to either largemouth bass chemical cues alone or chemical cues with diet cues (largemouth bass fed bullfrog tadpoles). To test whether this behavioral response was generalized across fish predators, we exposed tadpoles to rainbow trout ( Oncorhynchus mykiss) and found that tadpoles could not recognize this novel predator regardless of co‐occurrence with other fish species. These results suggest that environment may be more important for predator recognition than evolutionary history for this invasive species, and individuals do not retain predator recognition or generalize across fish predators.     

Predation risk in tadpole populations shapes behavioural responses of prey but not strength of trait‐mediated indirect interactions

Prey animals often respond to predators by reducing activity levels. This can produce a trait‐mediated indirect interaction (TMII) between predators and prey resources, whereby reduced foraging by prey in the presence of a predator causes an increase in prey resources. TMIIs play important roles in structuring communities, and it is important to understand factors that determine their strength. One such influence may be behavioural variation in the prey species, with indirect effects of predators being stronger within populations that are more responsive to the presence of a predator. We tested 1) whether the behavioural responsiveness of populations of wood frog tadpoles to predator cues was related to the predation risk in their native ponds, and 2) whether more responsive tadpoles yielded stronger TMIIs. To do this, we 1) measured the activity of tadpoles from 18 populations in mesocosms with and without caged predators, and 2) measured changes in the biomass of periphyton (the tadpoles’ diet) between predator treatments for each population. We found that tadpoles from higher predation risk ponds reduced their time outside refuges more in the presence of predators and tended to move less when visible, suggesting possible local adaptation to predation regimes. Though the presence of predators generally resulted in higher periphyton biomass – a TMII – there was no evidence that the strength of this TMII was affected by variation in tadpole behaviour. Foraging activity and general activity may be decoupled to some extent, enabling high predation risk‐adapted tadpoles to limit the fitness costs of reduced foraging when predators are present.     

Conspecific density determines the magnitude and character of predator-induced phenotype

The benefits in survival gained from predator-induced phenotypes often come at a cost to other components of fitness. Therefore, the level of expression of an induced phenotype should mirror the level of risk in the environment. When a predator exhibits a saturating functional response the risk of mortality to a given prey decreases as prey density increases. Therefore, for a given predator threat, investment in defense should be lower in prey at high density relative to those at low density. In this study, I test whether the magnitude of predator-induced morphological plasticity decreases with increasing conspecific density by exposing pine woods tree frog (Hyla femoralis) tadpoles at three different densities to predators (present or absent) in a factorial experiment. Tadpole morphology was not affected by changes in density in the absence of predators. However, predators had a significant, density-dependent effect on tadpole morphology. Specifically, the magnitude of morphological response was graded and larger for animals in the low density (high risk) environment. This study demonstrates that tadpoles can modulate phenotypic plasticity in response to mortality risk as a function of both the density of conspecifics and chemical cues from predators, which suggests that they are able to detect and respond to fine-scale changes in the threat environment. In addition, this study highlights the need for analytical approaches that allow morphological plasticity studies to elucidate allometric relationships in addition to simply quantifying size-corrected traits. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Prey detection of aquatic predators: Assessing the identity of chemical cues eliciting prey behavioral plasticity

Chemical cues transmitted through the environment are thought to underlie many prey responses to predation risk, but despite the known ecological and evolutionary significance of such cues, their basic composition are poorly understood. Using anuran tadpoles (prey) and dragonfly larvae (predators), we identified chemical cues associated with predation risk via solid phase extraction and mass spectrometry of the extracts. We found that dragonfly larvae predators consistently produced a negative ion, m/z 501.3, when they fed on bullfrog (Rana catesbeiana) and mink frog (Rana septentrionalis) tadpoles, but this ion was absent when dragonflies were fasted or fed invertebrate prey. When tadpole behavioral responses to dragonfly chemical cues were examined, tadpoles reduced their activity, particularly in response to dragonflies feeding on tadpoles. Furthermore, a negative correlation was noted between the level of tadpole activity and the concentration of the m/z 501.3 compound in dragonfly feeding trials, indicating that this ion was possibly responsible for tadpole anti-predator behavior.