Invasive Cane Toads Research Articles

Rapid adaptation to cold in the invasive cane toad Rhinella marina.

Understanding rapid adaptation to novel environments is essential as we face increasing climatic change. Invasive species are an ideal system for studying adaptation as they are typically introduced to novel environments where they must adapt if they are to persist. We used the invasive cane toad, Rhinella marina, to investigate the contribution of plasticity and evolution to rapid adaptation in a novel environment. Rhinella marina is a neotropical toad that has invaded areas with climates outside of its native environmental niche. The goal of this research was to understand how cane toads persist in northern Florida, the coldest region of their combined natural and invasive range, and originally thought to be beyond their thermal breadth. We measured Critical thermal minima in cane toads from the original, warm introduction location (Miami), and their northern range edge (Tampa) to determine whether northern toads were more cold-tolerant, and to examine the contribution of adaptive plasticity and evolution to any changes in tolerance. Our results show that following acclimation to cold temperatures, southern toads are less tolerant of cold than northern toads. This persistent population difference implies selection for cold-tolerance in northern populations. Differences in individual responses indicate that plasticity is also involved in this response. Our findings have implications for conservation because predatory cane toad invasions threaten local faunas, especially native amphibians. Characterizing specific adaptive mechanisms that allow R. marina to expand its range will identify evolutionary processes that shape a highly successful invasive species.

Ontogenetic shift in toxicity of invasive cane toads facilitates learned avoidance by native predators

Ontogenetic shift in toxicity of invasive cane toads facilitates learned avoidance by native predators

Age- and size-dependent resistance to chytridiomycosis in the invasive cane toad Rhinella marina.

In Australia, the cane toad Rhinella marina and chytrid fungus Batrachochytrium dendrobatidis (Bd) are examples of invasive species that have had dramatic impacts on native fauna. However, little is known about the interaction between Bd and cane toads. We aimed to explore the interaction of these 2 species in 3 parts. First, we collated data from the literature on Bd infection in wild cane toads. Second, we tested the susceptibility of recently metamorphosed cane toads to Bd infection. Finally, we modelled the distribution of the 2 species in Australia to identify where they overlap and, therefore, might interact. Through our data collation, we found that adult cane toads are infrequently infected and do not carry high infection burdens; however, our infection experiment showed that metamorphs are highly susceptible to infection and disease, but resistance appears to increase with increasing toad size. Niche modelling revealed overlapping distributions and the potential for cane toads to be affected by chytridiomycosis in the wild. While Bd can cause mortality in small juveniles in the laboratory, warm microhabitats used by wild toads likely prevent infection, and furthermore, high mortality of juveniles is unlikely to affect the adult populations because they are highly fecund. However, to demonstrate the impact of Bd on wild cane toad populations, targeted field studies are required to assess (1) the overall impact of chytridiomycosis on recruitment especially in cooler areas more favourable for Bd and (2) whether cane toad juveniles can amplify Bd exposure of native amphibian species in these areas.

Influence of a mortality trade-off on the spreading rate of cane toads fronts

We study the influence of a mortality trade-off in a nonlocal reaction-diffusion-mutation equation that we introduce to model the invasion of cane toads in Australia. This model is built off of one that has attracted attention recently, in which the population of toads is structured by a phenotypical trait that governs the spatial diffusion. We are concerned with the case when the diffusivity can take unbounded values and the mortality trade-off depends only on the trait variable. Depending on the rate of increase of the penalization term, we obtain the rate of spreading of the population. We identify two regimes, an acceleration regime when the penalization is weak and a linear spreading regime when the penalization is strong. While the development of the model comes from biological principles, the bulk of the article is dedicated to the mathematical analysis of the model, which is very technical. The upper and lower bounds are proved via the Li-Yau estimates of the fundamental solution of the heat equation with potential on Riemannian manifolds and a moving ball technique, respectively, and the traveling waves by a Leray-Schauder fixed point argument. We also present a simple method for a priori bounds.

Behavioural tactics used by invasive cane toads (Rhinella marina) to exploit apiaries in Australia

AbstractBehavioural flexibility plays a key role in facilitating the ability of invasive species to exploit anthropogenically‐created resources. In Australia, invasive cane toads (Rhinella marina) often gather around commercial beehives (apiaries), whereas native frogs do not. To document how toads use this resource, we spool‐tracked cane toads in areas containing beehives and in adjacent natural habitat without beehives, conducted standardized observations of toad feeding behaviour, and ran prey‐manipulation trials to compare the responses of cane toads versus native frogs to honeybees as potential prey. Toads feeding around beehives travelled shorter distances per night, and hence used different microhabitats, than did toads from nearby control sites without beehives. The toads consumed live bees from the hive entrance (rather than dead bees from the ground), often climbing on top of one another to gain access to the hive entrance. Prey manipulation trials confirm that bee movement is the critical stimulus that elicits the toads’ feeding response; and in standardized trials, native frogs consumed bees less frequently than did toads. In summary, cane toads flexibly modify their movements, foraging behaviour and dietary composition to exploit the nutritional opportunities created by commercial beehives, whereas native anurans do not.

Taste overshadows less salient cues to elicit food aversion in endangered marsupial

Conditioned taste aversion is an emerging conservation tool that can be used to limit inter-species conflict, for example decreasing predation on endangered species, or limiting the consumption of invasive toxic prey. Typically, managers wish to elicit an aversion that will be associated with visual or odour cues so that the target species maintains an “arm’s length” relationship, and does not have to attack or taste the prey. Combining multiple cues in conditioned taste aversion can cause cue overshadowing, so reducing the effectiveness of the training. Here, we examine the northern quoll (Dasyurus hallucatus), a carnivorous marsupial threatened because they attack the toxic invasive cane toad (Rhinella marina). Conditioned taste aversion has been suggested as a way to reduce quoll’s predation on toads, but for training to be effective it must elicit an aversion to stimuli before the quolls attack the toad. Using baits containing distinct novel meat, odour and visual cues, we test whether quolls will generalise their aversion to visual or odour cues. We found that quolls associate their aversion with the compound stimulus of meat taste and meat odour, and that this overshadows the less salient artificial odour and visual cues. Quolls ate 87% of taste cue substituted baits, compared to 38% and 42% of the odour and visual cue substituted baits respectively, which represented a significant interaction between bait type and substituted cue (χ2(4) = 14.70, p = 0.005). We show that although quolls do not generalise their aversions to novel artificial stimuli, they can distinguish subtle visual and odour cues in meat that still elicit an aversion without them needing to attack and taste prey.

Environmental DNA sampling as a surveillance tool for cane toad Rhinella marina introductions on offshore islands

Containing the spread of established invasive species is critical for minimizing their ecological impact. Effective containment requires sensitive sampling methods capable of detecting new introductions when invaders are at low density. Here we explore whether environmental DNA (eDNA) sampling could be used as a surveillance tool to detect new incursions of aquatic invasive species on offshore islands. We develop an eDNA molecular assay for invasive cane toads (Rhinella marina) in Australia, validate our assay on the mainland, and apply it to an offshore island (Moreton Island) that is a target of ongoing cane toad surveillance. Our eDNA assay correctly identified four mainland sites at which cane toads were observed, as well as a fifth site within 1 km of known populations. Five additional sites outside the cane toad’s current distribution tested negative for cane toad eDNA. Site occupancy detection models indicated that two water samples and three qPCR replicates were sufficient to achieve a cumulate detection probability > 0.95. Applying our eDNA assay to samples from 19 sites on an offshore island over a 2-year period revealed the absence of cane toad eDNA, in line with our current understanding of cane toad distribution. Our results suggest that eDNA sampling could be strategically applied to meet the Australian Commonwealth’s objective of maintaining cane toad-free offshore islands.

Draft genome assembly of the invasive cane toad, Rhinella marina.

BackgroundThe cane toad (Rhinella marina formerly Bufo marinus) is a species native to Central and South America that has spread across many regions of the globe. Cane toads are known for their rapid adaptation and deleterious impacts on native fauna in invaded regions. However, despite an iconic status, there are major gaps in our understanding of cane toad genetics. The availability of a genome would help to close these gaps and accelerate cane toad research.FindingsWe report a draft genome assembly for R. marina, the first of its kind for the Bufonidae family. We used a combination of long-read Pacific Biosciences RS II and short-read Illumina HiSeq X sequencing to generate 359.5 Gb of raw sequence data. The final hybrid assembly of 31,392 scaffolds was 2.55 Gb in length with a scaffold N50 of 168 kb. BUSCO analysis revealed that the assembly included full length or partial fragments of 90.6% of tetrapod universal single-copy orthologs (n = 3950), illustrating that the gene-containing regions have been well assembled. Annotation predicted 25,846 protein coding genes with similarity to known proteins in Swiss-Prot. Repeat sequences were estimated to account for 63.9% of the assembly.ConclusionsThe R. marina draft genome assembly will be an invaluable resource that can be used to further probe the biology of this invasive species. Future analysis of the genome will provide insights into cane toad evolution and enrich our understanding of their interplay with the ecosystem at large.

Differential gene expression to an LPS challenge in relation to exogenous corticosterone in the invasive cane toad (Rhinella marina)

The cane toad (Rhinella marina) is an invasive amphibian in several parts of the world. Much of the research performed on assessing the dispersal potential of invasive species has focused immunity. Invaders are predicted to rely less on pro-inflammatory immunity, allowing them to allocate energy to dispersal. Elevated stress may play a role in regulation of immune responses used by invasive species. RNA sequencing of spleen tissue from cane toads subjected to an acute LPS challenge revealed genes coding for cytokines involved in typical innate responses such as phagocytic cell recruitment, extravasation, inflammation, and lymphocyte differentiation were significantly upregulated, while toads receiving transdermal application of corticosterone in addition to an LPS injection showed downregulation of genes involved with cell mediated immunity. These results indicate hormonal changes associated with acute stress may alter investment into mounting cell-mediated or humoral responses while allowing for prolonged phagocytic innate responses in this invasive species.

Cloacal and Ocular Microbiota of the Endangered Australian Northern Quoll

The Australian northern quoll is an important predatory marsupial carnivore that is currently endangered due to inappropriate fire regimes, predation, and the spread of invasive cane toads. The microbiota of Australian marsupials has not been extensively studied, but is thought to play a role in their health. This study provides an initial characterization of the cloacal microbiota of the northern quoll, as well as other marsupials including possums and kangaroos which were opportunistically sampled. The northern quoll cloaca microbiota was dominated by Enterococcus and Lactobacillus and had a relatively high proportion of members of the Proteobacteria phylum, which has been observed in other carnivorous marsupials. The diversity and structure of the microbiota was not influenced by presence of Chlamydiales which are intracellular bacteria and potential pathogens. The microbiota of the other marsupials was quite varied, which may be related to their health status. Characterization of the northern quoll microbiota will help to better understand the biology of this endangered animal.

The perils of paradise: an endangered species conserved on an island loses antipredator behaviours within 13 generations.

When imperilled by a threatening process, the choice is often made to conserve threatened species on offshore islands that typically lack the full suite of mainland predators. While keeping the species extant, this releases the conserved population from predator-driven natural selection. Antipredator traits are no longer maintained by natural selection and may be lost. It is implicitly assumed that such trait loss will happen slowly, but there are few empirical tests. In Australia, northern quolls (Dasyurus hallucatus) were moved onto a predator-free offshore island in 2003 to protect the species from the arrival of invasive cane toads on the mainland. We compared the antipredator behaviours of wild-caught quolls from the predator-rich mainland with those from this predator-free island. We compared the responses of both wild-caught animals and their captive-born offspring, to olfactory cues of two of their major predators (feral cats and dingoes). Wild-caught, mainland quolls recognized and avoided predator scents, as did their captive-born offspring. Island quolls, isolated from these predators for only 13 generations, showed no recognition or aversion to these predators. This study suggests that predator aversion behaviours can be lost very rapidly, and that this may make a population unsuitable for reintroduction to a predator-rich mainland.

Sexual and geographical divergence in head widths of invasive cane toads, Rhinella marina (Anura: Bufonidae), is driven by both rapid evolution and plasticity

Sexual and geographical divergence in head widths of invasive cane toads, Rhinella marina (Anura: Bufonidae), is driven by both rapid evolution and plasticity

Effects of rearing environment and population origin on responses to repeated behavioural trials in cane toads (Rhinella marina)

Behavioural responses to repeated trials in captivity can be driven by many factors including rearing environment, population of origin, habituation to captivity/trial conditions and an individual’s behavioural type (e.g., bold versus shy). We tested the effect of rearing environment (captive raised common-garden versus wild-caught) and population origin (range-edge versus range-front) on the responses of invasive cane toads (Rhinella marina) to repeated exploration and risk-taking assays in captivity. We found that behavioural responses to identical assays performed on two occasions were complex and showed few consistent patterns based on rearing environment or population of origin. However, behavioural traits were repeatable across Trial Blocks when all sample populations were grouped together, indicating general consistency in individual toad behaviour across repeated behavioural assays. Our findings exemplify the complexity and unpredictability of behavioural responses and their effects on the repeatability and interpretation of behavioural traits across repeated behavioural assays in captivity. To meaningfully interpret the results from repeated behavioural assays, we need to consider how multiple factors may affect behavioural responses to these tests and importantly, how these responses may affect the repeatability of behavioural traits across time.

An evaluation of methods used to cull invasive cane toads in tropical Australia

To identify cost-effective ways to control invasive species, we need to evaluate alternative methods. The invasion of tropical Australia by cane toads (Rhinella marina) has killed many native predators, prompting efforts to cull adult toads. We analyzed a dataset on > 17,500 toads killed by government-employed teams from 2005 to 2008, using a range of methods (incidental captures, targeted searches, traps) as well as records of toads found as road kills. The culling occurred in the northwestern part of the Northern Territory, moving west as the toad front expanded into Western Australia. Both season and method affected rates of capture and demographic attributes (sex ratio, proportion of adults) of the culled anurans, with a strong interaction between these factors. Most methods were more effective during and after the seasonal monsoons (when moist conditions facilitated anuran activity), but that seasonal variation was greater for numbers of toads found on the roads than captured in other ways. Juvenile toads and adult female toads constituted higher proportions of total captures in the Early Dry than in other times of year, reflecting seasonal breeding. Traps caught a higher proportion of adult female toads than did other methods, but overall had low capture rates. Rates of range expansion by toads were similar before, during and after the culling effort, suggesting that removing adult toads did not affect the speed of the invasion. Nonetheless, culling adult cane toads may be effective in some situations (“island” populations, extralimital translocations), and results from this program may be valuable for future toad-control efforts.

Behavioural responses of an Australian colubrid snake (Dendrelaphis punctulatus) to a novel toxic prey item (the Cane Toad Rhinella marina)

The invasion of a toxic prey type can differentially affect closely related predator species. In Australia, the invasive Cane Toad (Rhinella marina) kills native anurophagous predators that cannot tolerate the toad’s toxins; but predators that are physiologically resistant (i.e., belong to lineages that entered Australia recently from Asia, where toads of other species are common) have been more resilient. In the current study, we examine the case of an Asian-derived predator lineage that relies on behavioural not physiological adaptations to deal with toads. Despite their Asian origins, Common Tree Snakes (Dendrelaphis punctulatus) are highly sensitive to toad toxins; yet this snake has not declined in abundance due to toads. We exposed captive (field-collected) snakes to toads of different sizes and ontogenetic stages, to quantify feeding responses and outcomes. Tree Snakes were less likely to attack toads than to attack native frogs, and rarely retained their hold on large toads. Tree Snakes ingested frogs of a wide range of body sizes but only ingested very small toads (< 1 g vs. up to 30 g for frogs). Behavioural responses were virtually identical between Tree Snakes from invaded versus yet-to-be-invaded areas, suggesting that preadaptation (from Asia) rather than adaptation (within Australia) is the key to successful utilisation of this novel but potentially toxic prey resource. Nonetheless, a previously-documented shift in relative head sizes of Tree Snakes coincident with toad invasion suggests that the ancestral behavioural tactic may have been reinforced by a recent morphological shift that further reduces maximal prey size, and hence the risk of fatal poisoning.

Not such silly sausages: Evidence suggests northern quolls exhibit aversion to toads after training with toad sausages

AbstractThe invasion of toxic cane toads (Rhinella marina) is a major threat to northern quolls (Dasyurus hallucatus) which are poisoned when they attack this novel prey item. Quolls are now endangered as a consequence of the toad invasion. Conditioned taste aversion can be used to train individual quolls to avoid toads, but we currently lack a training technique that can be used at a landscape scale to buffer entire populations from toad impact. Broad‐scale deployment requires a bait that can be used for training, but there is no guarantee that such a bait will ultimately elicit aversion to toads. Here, we test a manufactured bait – a ‘toad sausage’ – in a small captive trial, for its ability to elicit aversion to toads in northern quolls. To do this, we exposed one group of quolls to a toad sausage and another to a control sausage and compared the quolls' predatory responses when presented with a dead adult toad. Captive quolls that consumed a single toad sausage showed a reduced interest in cane toads, interacting with them for less than half the time of their untrained counterparts and showing reduced Attack behaviour. We also quantified bait uptake in the field, by both quolls and non‐target species. These field trials showed that wild quolls were the most frequent species attracted to the baits, and that approx. 61% of quolls consumed toad‐aversion baits when first encountered. Between 40% and 68% of these animals developed aversion to further bait consumption. Our results suggest that toad‐aversion sausages may be used to train wild quolls to avoid cane toads. This opens the possibility for broad‐scale quoll training with toad aversion sausages: a technique that may allow wildlife managers to prevent quoll extinctions at a landscape scale.

Invasion history alters the behavioural consequences of immune system activation in cane toads.

Acute activation of the immune system often initiates a suite of behavioural changes. These "sickness behaviours"-involving lethargy and decreased activity-may be particularly costly on invasion fronts, where evolutionary pressures on dispersal favour individuals that move large distances. We used a combination of field and laboratory studies to compare sickness behaviours of cane toads from populations differing in invasion history. To do this we stimulated immune system activation by injecting lipopolysaccharide (LPS) to mimic bacterial infection. We predicted that LPS would result in less severe sickness behaviour in toads from range-edge populations because they had undergone selection for rapid and sustained dispersal (activities in conflict with lethargy and decreased activity). Contrary to our prediction, LPS injection caused a greater reduction in dispersal-relevant traits in invasion-front individuals than in conspecifics from the range-core. Our data suggest that the rapid invasion of cane toads through tropical Australia has seen an evolutionary shift in the magnitude of sickness behaviour elicited by pathogen infection. The increased sickness behaviour among range-edge toads suggests a shift away from pathogen tolerance (seen in range-core populations) towards resistance to pathogen attack. But as a consequence, when pathogens do become successfully established, toads from invasion-front populations may have less capacity to tolerate their ill-effects.

The establishment and eradication of an extra-limital population of invasive cane toads

Biological invasions expand not only as a continuous front, but also via translocation of small numbers of individuals to sites far from the main distribution. Detecting and eradicating such satellite populations is critical to curtailing the invasion. Cane toads (Rhinella marina) have been spreading through Australia since 1935, and in 2010 were reported in suburban Sydney, 500 km from the main invasion. The amphibians arrived as stowaways in trucks, and bred successfully in at least 3 years. Local residents noted the toads’ presence long before management authorities. As soon as they recognised the population’s establishment, however, the authorities surveyed locations of toads, assessed toad demography, and culled toads. Radio-tracking revealed the locations of spawning sites, which were then fenced to exclude access. As a result of reproduction, growth, and sex-specific culling (due to sex differences in distribution and catchability), successive years saw major changes in toad abundance, body-size distributions and sex ratios. Rates of infection of toads by parasites (lungworms) increased through time also. Adult male toads were collected primarily from the initial introduction site, whereas adult females were collected from a broader area. A total of > 900 post-metamorphic cane toads and > 5000 earlier life stages were collected, and those efforts (plus predation by rats and birds) exterminated the population (no sightings for the past 3 years). Cane toads will continue to be translocated to sites far outside their main invaded range, but the successful eradication effort suggests that well-integrated programs, with close collaboration among stakeholders, can prevent such populations from establishing.

Invasive cane toads might initiate cascades of direct and indirect effects in a terrestrial ecosystem

Understanding the impacts that invasive vertebrates have on terrestrial ecosystems extends primarily to invaders’ impacts on species with which they interact directly through mechanisms such as predation, competition and habitat modification. In addition to direct effects, invaders can also initiate ecological cascades via indirect population level effects on species with which they do not directly interact. However, evidence that invasive vertebrates initiate ecological cascades in terrestrial ecosystems remains scarce. Here, we ask whether the invasion of the cane toad, a vertebrate invader that is toxic to many of Australia’s vertebrate predators, has induced ecological cascades in a semi-arid rangeland. We compared activity of a large predatory lizard, the sand-goanna, and abundances of smaller lizards preyed upon by goannas in areas of high toad activity near toads’ dry season refuges and areas of low toad activity distant from toads’ dry season refuges. Consistent with the hypothesis that toad invasion has led to declines of native predators susceptible to poisoning, goanna activity was lower in areas of high toad activity. Consistent with the hypothesis that toad-induced goanna decline lead to increases in abundance the prey of goannas, smaller lizards were more abundant in areas of high toad activity. Structural equation modelling showed a positive correlation between goanna activity and distance from dry season refuge habitats used by toads. The abundances of small lizards was correlated negatively with goanna activity and distance from dry season refuges of toads. Our findings provide support for the notion that invasions by terrestrial vertebrates can trigger ecological cascades.

Physiological plasticity in a successful invader: rapid acclimation to cold occurs only in cool-climate populations of cane toads (Rhinella marina).

Physiological plasticity may facilitate invasion of novel habitats; but is such plasticity present in all populations of the invader or is it elicited only by specific climatic challenges? In cold-climate areas of Australia, invasive cane toads (Rhinella marina) can rapidly acclimate to cool conditions. To investigate whether this physiological plasticity is found in all invasive cane toads or is only seen in cool climates, we measured the acclimation ability of toads from across Australia and the island of Hawai'i. We collected toads from the field and placed them at either 12 or 24°C for 12 h before measuring their righting response as a proxy for critical thermal minimum (CTmin). Toads from the coolest Australian region (New South Wales) demonstrated plasticity (as previously reported), with exposure to 12°C (vs. 24°C) decreasing CTmin by 2°C. In toads from other Australian populations, CTmins were unaffected by our thermal treatments. Hawai'ian toads from a cool, wet site also rapidly acclimated to cool conditions, whereas those from warmer and drier Hawai'ian sites did not. Thermal plasticity has diverged among populations of invasive cane toads, with rapid acclimation manifested only in two cool-climate populations from widely separated sites. Predictions about the potential range of invasive species thus must consider the possibility of geographic (intraspecific) heterogeneity in thermal plasticity; data from other parts of the species' range may fail to predict levels of plasticity elicited by thermal challenges.