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Considerable research has focused on microbes on amphibian skin, as they act as the first line of defense against invading pathogens. This effort has generated substantial data on patterns across species, space, time, and ontogeny, alongside a growing list of beneficial antifungal symbionts. Though there is evidence of stability in amphibian skin microbial communities, there is also an indication that regular skin shedding reduces cultivable bacteria, with regrowth and recolonization in the period between sheds. This suggests that skin communities are in constant flux, and we lack an understanding of how the membership and structure of those communities are affected by shedding events. In this study, we conducted experiments on cane toads (Rhinella marina) to investigate the influence of shedding on skin microbiomes. We first used quantitative PCR to verify a positive correlation between bacterial loads and time in the days after shedding. We then resampled individuals over time to describe changes in community composition in the 38 h after shedding using amplicon sequencing. Similar to trends of bacterial loads, we found increases in alpha diversity over time after shedding, suggesting that shedding reduces bacterial diversity as it knocks down bacterial loads. During the 38-h period, community structure became similar to pre-shed communities in some individuals, but there was no consistent pattern in structural changes among individuals. In light of the amphibian chytridiomycosis pandemic, understanding how physiological events such as skin shedding affect beneficial bacteria and communities on amphibians would provide important insight into amphibian ecology., (© 2024 The Author(s). Integrative Zoology published by International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

Many organisms can adjust their development according to environmental conditions, including the presence of conspecifics. Although this developmental plasticity is common in amphibians, its underlying molecular mechanisms remain largely unknown. Exposure during development to either 'cannibal cues' from older conspecifics, or 'alarm cues' from injured conspecifics, causes reduced growth and survival in cane toad ( Rhinella marina ) tadpoles. Epigenetic modifications, such as changes in DNA methylation patterns, are a plausible mechanism underlying these developmental plastic responses. Here we tested this hypothesis, and asked whether cannibal cues and alarm cues trigger the same DNA methylation changes in developing cane toads. We found that exposure to both cannibal cues and alarm cues was associated with local changes in DNA methylation patterns. These DNA methylation changes affected genes putatively involved in developmental processes, but in different genomic regions for different conspecific-derived cues. Genetic background explains most of the epigenetic variation among individuals. Overall, the molecular mechanisms triggered by exposure to cannibal cues seem to differ from those triggered by alarm cues. Studies linking epigenetic modifications to transcriptional activity are needed to clarify the proximate mechanisms that regulate developmental plasticity in cane toads., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

By imposing novel selection pressures on both participants, biological invasions can modify evolutionary 'arms races' between hosts and parasites. A spatially replicated cross-infection experiment reveals strong spatial divergence in the ability of lungworms ( Rhabdias pseudosphaerocephala ) to infect invasive cane toads ( Rhinella marina ) in Australia. In areas colonized for longer than 20 years, toads are more resistant to infection by local strains of parasites than by allopatric strains. The situation reverses at the invasion front, where super-infective parasites have evolved. Invasion-induced shifts in genetic diversity and selective pressures may explain why hosts gain advantage over parasites in long-colonized areas, whereas parasites gain advantage at the invasion front.

In Canada and internationally the number of older offenders on parole and living in the community is increasing rapidly. Older offenders in the community are a vulnerable population at high risk for lack of well-being. Semi-structured interviews were conducted with N  = 64 offenders aged 50 years and older on conditional release from custody in Canada, including long-term, recidivist, and first-time older offenders. Compared to their non-offender counterparts, older offenders in the community experience many of the same problems of aging and well-being, but are at greater risk for mental health problems, traumatic injuries, and for recidivists, substance abuse. Most long-term and first-time older offenders find themselves living at or below the poverty line. One third of older offenders experience social isolation from community and family due to their criminal history and incarceration. For those with Indigenous ancestry, Indigenous communities, and cultural organizations play a significant role in supporting older offenders., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Evolutionary shifts from one habitat type to another can clarify selective forces that affect life-history attributes. Four lineages of snakes (acrochordids and three clades within the Elapidae) have invaded marine habitats, and all have larger offspring than do terrestrial snakes. Predation by fishes on small neonates offers a plausible selective mechanism for that shift, because ascending to breathe at the ocean surface exposes a marine snake to midwater predation whereas juvenile snakes in terrestrial habitats can remain hidden. Consistent with this hypothesis, snake-shaped models moving through a coral-reef habitat in New Caledonia attracted high rates of attack by predatory fishes, and small models (the size of neonatal terrestrial snakes) were attacked more frequently than were large models (the size of neonatal sea snakes). Vulnerability to predatory fishes may have imposed strong selection for increased offspring size in marine snakes., Competing Interests: We declare we have no competing interests., (© 2023 The Authors.)

Variation in food resources can result in dramatic fluctuations in the body condition of animals dependent on those resources. Decreases in body mass can disrupt patterns of energy allocation and impose stress, thereby altering immune function. In this study, we investigated links between changes in body mass of captive cane toads ( Rhinella marina ), their circulating white blood cell populations, and their performance in immune assays. Captive toads that lost weight over a three-month period had increased levels of monocytes and heterophils and reduced levels of eosinophils. Basophil and lymphocyte levels were unrelated to changes in mass. Because individuals that lost mass had higher heterophil levels but stable lymphocyte levels, the ratio of these cell types was also higher, partially consistent with a stress response. Phagocytic ability of whole blood was higher in toads that lost mass, owing to increased circulating levels of phagocytic cells. Other measures of immune performance were unrelated to mass change. These results highlight the challenges faced by invasive species as they expand their range into novel environments which may impose substantial seasonal changes in food availability that were not present in the native range. Individuals facing energy restrictions may shift their immune function towards more economical and general avenues of combating pathogens. This article is part of the theme issue 'Amphibian immunity: stress, disease and ecoimmunology'.

Bacterial assemblages on amphibian skin may play an important role in protecting hosts against infection. In hosts that occur over a range of environments, geographic variation in composition of bacterial assemblages might be due to direct effects of local factors and/or to evolved characteristics of the host. Invasive cane toads (Rhinella marina) are an ideal candidate to evaluate environmental and genetic mechanisms, because toads have evolved major shifts in physiology, morphology, and behavior during their brief history in Australia. We used samples from free-ranging toads to quantify site-level differences in bacterial assemblages and a common-garden experiment to see if those differences disappeared when toads were raised under standardised conditions at one site. The large differences in bacterial communities on toads from different regions were not seen in offspring raised in a common environment. Relaxing bacterial clustering to operational taxonomic units in place of amplicon sequence variants likewise revealed high similarity among bacterial assemblages on toads in the common-garden study, and with free-ranging toads captured nearby. Thus, the marked geographic divergence in bacterial assemblages on wild-caught cane toads across their Australian invasion appears to result primarily from local environmental effects rather than evolved shifts in the host., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Repeat contacts between persons with serious mental disorder (PSMD) and police officers are costly to both police services and hospitals in terms of service utilization and can be viewed as a direct indicator of unmet needs and gaps in service provision. The intent of the current study was to examine the demographic and clinical characteristics of PSMD who had repeat contact with police officers in London, Ontario from 2016 to 2019 using data collected using the interRAI Brief Mental Health Screener (BMHS). Negative binomial regression was used on a sample of 4143 cases to develop a model predictive of repeat police contacts. The most parsimonious model predicting police contact based on items on the BMHS included age, command hallucinations, lack of insight, verbal abuse, known to possess weapons and family, friends and caregivers expressing concern over the possibility of self harm. Delusions were also independently significantly associated with repeat encounters. Unique to this study is the observation that possessing a weapon in the past 12 months was included in the predictive model. The results of the study add to the sparse research devoted to identifying the characteristics of PSMD who have repeat contact with police officers. Recommendations include integrating the findings into police training to ensure police officers flag those who have the potential for repeat encounters and refer them to appropriate community mental health service providers for proactive outreach services., (Copyright © 2022. Published by Elsevier Ltd.)

Comparative studies of mortality in the wild are necessary to understand the evolution of aging; yet, ectothermic tetrapods are underrepresented in this comparative landscape, despite their suitability for testing evolutionary hypotheses. We present a study of aging rates and longevity across wild tetrapod ectotherms, using data from 107 populations (77 species) of nonavian reptiles and amphibians. We test hypotheses of how thermoregulatory mode, environmental temperature, protective phenotypes, and pace of life history contribute to demographic aging. Controlling for phylogeny and body size, ectotherms display a higher diversity of aging rates compared with endotherms and include phylogenetically widespread evidence of negligible aging. Protective phenotypes and life-history strategies further explain macroevolutionary patterns of aging. Analyzing ectothermic tetrapods in a comparative context enhances our understanding of the evolution of aging.

Life-history traits such as rates of growth, survival and reproduction can vary though time within a single population, or through space among populations, due to abiotically-driven changes in resource availability. In terrestrial reptiles, parameters such as temperature and rainfall generate variation in life-histories-but other parameters likely are more important in marine systems. We studied three populations of sea snakes (Emydocephalus annulatus) in adjacent bays in the IndoPacific archipelago of New Caledonia. The extreme philopatry of individual snakes allows us to unambiguously allocate each animal to one of the three populations. Although water temperatures and rainfall do not differ over this small scale, one site experiences more intense winds, restricting opportunities for foraging. Our 18-year mark-recapture dataset (> 1,200 snakes, > 2,400 captures) reveals significant divergence among populations in life-history traits. Survival rates and population densities were similar among sites, but snakes at the most wind-exposed site (Anse Vata) exhibited lower body condition, slower growth, less frequent production of litters, and smaller litters. Weather-driven variation in feeding rates thus may affect life-history traits of marine snakes as well as their terrestrial counterparts, but driven by different parameters (e.g., wind exposure rather than variation in temperatures or rainfall)., (© 2022. The Author(s).)

Evolutionary theory suggests that polymorphic traits can be maintained within a single population only under specific conditions, such as negative frequency-dependent selection or heterozygote advantage. Non-venomous turtle-headed sea snakes (Emydocephalus annulatus) living in shallow bays near Noumea in New Caledonia exhibit three colour morphs: black, black-and-white banded, and an intermediate (grey-banded) morph that darkens with age. We recorded morph frequencies during 18 consecutive years of surveys, and found that the numbers of recruits (neonates plus immigrants) belonging to each morph increased in years when that morph was unusually rare in the population, and decreased when that morph was unusually common. Thus, morph frequencies are maintained by negative frequency-dependent selection. We interpret the situation as Batesian mimicry of highly venomous sea snakes (Aipysurus, Hydrophis, Laticauda) that occur in the same bays, and range in colour from black-and-white banded to grey-banded. Consistent with the idea that mimicry may protect snakes from attack by large fish and sea eagles, behavioural studies have shown that smaller fish species in these bays flee from banded snakes but attack black individuals. As predicted by theory, mimetic (banded) morphs are less common than the cryptically-coloured melanic morph., (© 2022. The Author(s).)

Evolutionary arms races can alter both parasite infectivity and host resistance, and it is difficult to separate the effects of these twin determinants of infection outcomes. We used a co-introduced, invasive host-parasite system (the lungworm Rhabdias pseudosphaerocephala and cane toads Rhinella marina ), where rapid adaptation and dispersal have led to population differences in infection resistance. We quantified behavioural responses of parasite larvae to skin-chemical cues of toads from different invasive populations, and rates at which juvenile hosts became infected following standardized exposure to lungworms. Chemical cues from toad skin altered host-seeking behaviour by parasites, similarly among populations. The number of infection attempts (parasite larvae entering the host's body) also did not differ between populations, but rates of successful infection (establishment of adult worm in host lungs) were higher for range-edge toads than for range-core conspecifics. Thus, lower resistance to parasite infection in range-edge juvenile toads appears to be due to less effective immune defences of the host rather than differential behavioural responses of the parasite. In this ongoing host-parasite arms race, changing outcomes appear to be driven by shifts in host immunocompetence.

Invasions often accelerate through time, as dispersal-enhancing traits accumulate at the expanding range edge. How does the dispersal behaviour of individual organisms shift to increase rates of population spread? We collate data from 44 radio-tracking studies (in total, of 650 animals) of cane toads (Rhinella marina) to quantify distances moved per day, and the frequency of displacement in their native range (French Guiana) and two invaded areas (Hawai'i and Australia). We show that toads in their native-range, Hawai'i and eastern Australia are relatively sedentary, while toads dispersing across tropical Australia increased their daily distances travelled from 20 to 200 m per day. That increase reflects an increasing propensity to change diurnal retreat sites every day, as well as to move further during each nocturnal displacement. Daily changes in retreat site evolved earlier than did changes in distances moved per night, indicating a breakdown in philopatry before other movement behaviours were optimised to maximise dispersal., (© 2021. The Author(s).)

For sea snakes as for many types of animals, long-term studies on population biology are rare and hence, we do not understand the degree to which annual variation in population sizes is driven by density-dependent regulation versus by stochastic abiotic factors. We monitored three populations of turtle-headed sea snakes (Emydocephalus annulatus) in New Caledonia over an 18-year period. Annual recruitment (% change in numbers) showed negative density-dependence: that is, recruitment increased when population densities were low, and decreased when densities were high. Windy weather during winter increased survival of neonates, perhaps by shielding them from predation; but those same weather conditions reduced body condition and the reproductive output of adult snakes. The role for density-dependence in annual dynamics of these populations is consistent with the slow, K-selected life-history attributes of the species; and the influence of weather conditions on reproductive output suggests that females adjust their allocation to reproduction based on food availability during vitellogenesis., (© 2021. The Author(s).)

Understanding how animal populations respond to environmental factors is critical because large-scale environmental processes (e.g., habitat fragmentation, climate change) are impacting ecosystems at unprecedented rates. On an overgrazed floodplain in north-western Australia, a native rodent (Pale Field Rat, Rattus tunneyi) constructs its burrows primarily beneath an invasive tree (Chinee Apple, Ziziphus mauritiana) rather than native trees. The dense thorny foliage of the Chinee Apple may allow high rat densities either because of abiotic effects (shade, in a very hot environment) or biotic processes (protection from trampling and soil compaction by feral horses, and/or predation). To distinguish between these hypotheses, we manipulated Chinee Apple foliage to modify biotic factors (access to horses and predators) but not shade levels. We surveyed the rat population with Elliott traps under treatment and control trees and in the open woodland, in two seasons (the breeding season-January, and the nesting season-May). In the breeding season, we ran giving-up density experiments (GUD) with food trays, to assess the perceived risk of predation by rats across our three treatments. Selective trimming of foliage did not affect thermal regimes underneath the trees but did allow ingress of horses and we observed two collapsed burrows as a consequence (although long term impacts of horses were not measured). The perceived predation risk also increased (GUD values at food trays increased) and was highest in the open woodland. Our manipulation resulted in a shift in rat sex ratios (indicating female preference for breeding under control but not foliage-trimmed trees) and influenced rat behaviour (giving-up densities increased; large dominant males inhabited the control but not treatment trees). Our data suggest that the primary benefit of the Chinee Apple tree to native rodents lies in physical protection from predators and (potentially) feral horses, rather than in providing cooler microhabitat.

In response to novel environments, invasive populations often evolve rapidly. Standing genetic variation is an important predictor of evolutionary response but epigenetic variation may also play a role. Here, we use an iconic invader, the cane toad ( Rhinella marina ), to investigate how manipulating epigenetic status affects phenotypic traits. We collected wild toads from across Australia, bred them, and experimentally manipulated DNA methylation of the subsequent two generations (G1, G2) through exposure to the DNA methylation inhibitor zebularine and/or conspecific tadpole alarm cues. Direct exposure to alarm cues (an indicator of predation risk) increased the potency of G2 tadpole chemical cues, but this was accompanied by reductions in survival. Exposure to alarm cues during G1 also increased the potency of G2 tadpole cues, indicating intergenerational plasticity in this inducible defence. In addition, the negative effects of alarm cues on tadpole viability (i.e. the costs of producing the inducible defence) were minimized in the second generation. Exposure to zebularine during G1 induced similar intergenerational effects, suggesting a role for alteration in DNA methylation. Accordingly, we identified intergenerational shifts in DNA methylation at some loci in response to alarm cue exposure. Substantial demethylation occurred within the sodium channel epithelial 1 subunit gamma gene ( SCNN1G ) in alarm cue exposed individuals and their offspring. This gene is a key to the regulation of sodium in epithelial cells and may help to maintain the protective epidermal barrier. These data suggest that early life experiences of tadpoles induce intergenerational effects through epigenetic mechanisms, which enhance larval fitness. This article is part of the theme issue 'How does epigenetics influence the course of evolution?'

Functional roles of the rich microbiota of the skin are not fully understood, but include protection against microbial diseases and other environmental challenges. In experimental studies, we show that reducing the microbiota from cane toad ( Rhinella marina ) skin by gently wiping with absorptive gauze resulted in threefold higher rates of infection by lungworms ( Rhabdias pseudosphaerocephala ) following standardised exposure to infective skin-penetrating larvae. Higher concentrations of microbial DNA were associated with lower rates of lungworm entry. Our data suggest that microbial activity on the anuran skin comprises an important line of defence against attack by macroparasites as well as by fungi and other microbes., Competing Interests: The authors declare that they have no conflict of interest., (© 2021 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Anthropogenic activities often create distinctive but discontinuously distributed habitat patches with abundant food but high risk of predation. Such sites can be most effectively utilized by individuals with specific behaviors and morphologies. Thus, a widespread species that contains a diversity of sizes and behavioral types may be pre-adapted to exploiting such hotspots. In eastern Australia, the giant (to >2 m) lizard Varanus varius (lace monitor) utilizes both disturbed (campground) and undisturbed (bushland) habitats. Our surveys of 27 sites show that lizards found in campgrounds tended to be larger and bolder than those in adjacent bushland. This divergence became even more marked after the arrival of a toxic invasive species (the cane toad, Rhinella marina ) caused high mortality in larger and bolder lizards. Some of the behavioral divergences between campground and bushland lizards may be secondary consequences of differences in body size, but other habitat-associated divergences in behavior are due to habituation and/or nonrandom mortality., Competing Interests: None declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

If optimal investment in anti-predator defences depends on predation risk, invading new regions (and thus, encountering different predators) may favour shifts in that investment. Cane toads offer an ideal system to test this prediction: expensive anti-predator toxins are stored mainly in parotoid glands whose dimensions are easy to measure, and toad invasions have changed the suites of predators they encounter. Although plasticity may influence parotoid morphology, comparisons between parents and progeny revealed that gland dimensions were highly heritable. That heritability supports the plausibility of an evolved basis to variation in gland dimensions. Measurements of 3779 adult toads show that females have larger glands than males, invasive populations have larger glands than in the native-range, and that parotoid sexual size dimorphism varies strongly among invaded areas. Geographic variation in parotoid morphology may be driven by predation risk to both adult toads and offspring (provisioned with toxins by their mother), with toxins allocated to eggs exacerbating the risk of cannibalism but reducing the risk of interspecific predation. Investment into chemical defences has evolved rapidly during the cane toad's international diaspora, consistent with the hypothesis that organisms flexibly adjust resource allocation to anti-predator tactics in response to novel challenges.

The water-permeable skin of amphibians renders them highly sensitive to climatic conditions, and interspecific correlations between environmental moisture levels and rates of water exchange across the skin suggest that natural selection adapts hydroregulatory mechanisms to local challenges. How quickly can such mechanisms shift when a species encounters novel moisture regimes? Cutaneous resistance to water loss and gain in wild-caught cane toads ( Rhinella marina ) from Brazil, USA (Hawai'i) and Australia exhibited strong geographic variation. Cutaneous resistance was low in native-range (Brazilian) toads and in Hawai'ian populations (where toads were introduced in 1932) but significantly higher in toads from eastern Australia (where toads were introduced in 1935). Toads from recently invaded areas in western Australia exhibited cutaneous resistance to water loss similar to the native-range populations, possibly because toads are restricted to moist sites within this highly arid landscape. Rates of rehydration exhibited significant but less extreme geographic variation, being higher in the native range than in invaded regions. Thus, in less than a century, cane toads invading areas that impose different climatic challenges have diverged in the capacity for hydroregulation., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Amphibians in hot climates may be able to avoid high temperatures by controlling their rates of heating. In northern Australia, invasive cane toads (Rhinella marina) experience hot dry conditions in newly-colonized (western) sites but milder conditions in longer-occupied (eastern) sites. Under standardized conditions, toads from western sites heated less rapidly than did conspecifics from an eastern site. The availability of free water slowed heating rates of eastern but not western toads. Thus, the colonization of climatically extreme sites has been accompanied by a rapid shift in the toads' ability to remain cool under hot conditions, even when free water is not available.

Tidal cycles are known to affect the ecology of many marine animals, but logistical obstacles have discouraged behavioural studies on sea snakes in the wild. Here, we analyse a large dataset (1,445 observations of 126 individuals) to explore tidally-driven shifts in the behaviour of free-ranging turtle-headed sea snakes (Emydocephalus annulatus, Hydrophiinae) in the Baie des Citrons, New Caledonia. Snakes tended to move into newly-inundated areas with the rising tide, and became more active (e.g. switched from inactivity to mate-searching and courting) as water levels rose. However, the relative use of alternative habitat types was largely unaffected by tidal phase.

Infection due to Entamoeba spp. is known to cause serious disease in primates ( Entamoeba histolytica ) and snakes ( Entamoeba invadens ), but there are no detailed descriptions of the pathology associated with Entamoeba spp. infection in amphibians. In 2014, an outbreak of entamoebiasis associated with a novel species of Entamoeba induced clinical illness and poor body condition in free-ranging cane toads in Australia's Northern Territory. Here, we describe the gross pathology, histology, and clinical pathology linked to the outbreak. The study compared 25 toads with invasive entamoebiasis, defined as histologically visible amoebas within tissue, and 12 toads without invasive entamoebiasis. Grossly, affected toads had mild to marked congestion of colonic serosal vasculature, with variable thickening of the intestinal wall and serosanguineous to hemorrhagic colonic content. Histologically, invasive entamoebiasis manifested primarily as moderate to severe, variably hyperplastic to ulcerative colitis. The small intestine was affected in 10 of 25 toads, and 5 of 25 toads also had gastric lesions. Amoebas consistent in morphology with Entamoeba sp. were commonly intermingled with mucosal epithelium, frequently along the basement membrane, with deeper invasion into the superficial lamina propria in only 5 toads. Toads with invasive entamoebiasis had neutrophilia, monocytosis, and lymphopenia, and thus elevated neutrophil to lymphocyte ratios, suggestive of an inflammatory and/or stress leukogram.

Initial research on the spread of cane toads (Rhinella marina) through tropical Australia reported a high incidence of spinal arthritis (spondylosis) in toads at the invasion front (where toads disperse rapidly), but not in areas colonized earlier (where toads are more sedentary). The idea that spondylosis was a cost of rapid dispersal was challenged by wider spatial sampling which linked rates of spondylosis to hot (tropical) climates rather than to dispersal rates. Here, the authors of these competing interpretations collaborate to reinterpret the data. Our reanalysis supports both previous hypotheses; rates of spondylosis are higher in populations established by fast-dispersing toads, and are higher in tropical than in temperate environments; they are also higher in larger toads. The functional reason for climatic effects is unclear, but might involve effects on the soil-living bacteria involved in the induction of spondylosis; and/or may reflect higher movement (as opposed to dispersal) or more pronounced dry-season aggregation rates of toads in tropical conditions.

Many animals capable of deploying chemical defences are reluctant to use them, suggesting that synthesis of toxins imposes a substantial cost. Typically, such costs have been quantified by measuring the elevation in metabolic rate induced by toxin depletion (i.e. during replenishment of toxin stores). More generally, we might expect that toxin depletion will induce shifts in a broad suite of fitness-relevant traits. In cane toads ( Rhinella marina), toxic compounds that protect against predators and pathogens are stored in large parotoid (shoulder) glands. We used correlational and experimental approaches in field and laboratory settings to investigate impacts of toxin depletion on growth rate and behaviour in cane toads. In free-ranging toads, larger toxin stores were associated with smaller gonads and livers, suggesting energetic trade-offs between toxin production and both reproduction and energy metabolism. Experimental removal of toxin (by manually squeezing parotoid glands) reduced rates of growth in body mass in both captive and free-ranging toads. Radio tracking demonstrated that de-toxined toads dispersed more slowly than did control toads. Given that toxin stores in cane toads take several months to fully replenish, deploying toxin to repel a predator may impose a substantial cost, explaining why toads use toxin only as a final line of defence.

Host-pathogen associations change rapidly during a biological invasion and are predicted to impose strong selection on immune function. It has been proposed that the invader may experience an abrupt reduction in pathogen-mediated selection ("enemy release"), thereby favoring decreased investment into "costly" immune responses. Across plants and animals, there is mixed support for this prediction. Pathogens are not the only form of selection imposed on invaders; differences in abiotic environmental conditions between native and introduced ranges are also expected to drive rapid evolution. Here, we use RNA-Seq to assess the expression patterns of immune and environmentally associated genes in the cane toad ( Rhinella marina ) across its invasive Australian range. Transcripts encoding mediators of costly immune responses (inflammation, cytotoxicity) showed a curvilinear relationship with invasion history, with highest expression in toads from oldest and newest colonized areas. This pattern is surprising given theoretical expectations of density dynamics in invasive species and may be because density influences both intraspecific competition and parasite transmission, generating conflicting effects on the strength of immune responses. Alternatively, this expression pattern may be the result of other evolutionary forces, such as spatial sorting and genetic drift, working simultaneously with natural selection. Our findings do not support predictions about immune function based on the enemy release hypothesis and suggest instead that the effects of enemy release are difficult to isolate in wild populations, especially in the absence of information regarding parasite and pathogen infection. Additionally, expression patterns of genes underlying putatively environmentally associated traits are consistent with previous genetic studies, providing further support that Australian cane toads have adapted to novel abiotic challenges., Competing Interests: None declared.

Dehydration is considered a physiological challenge, and many organisms live in environments that undergo periods of reduced water availability that can lead to dehydration. Recent studies have found a positive relationship between dehydration and innate immune function in animals adapted to xeric or semixeric environments. To explore the generality of this relationship, we examined the impact of dehydration on innate immune performance in water pythons (Liasis fuscus), a semiaquatic snake from the wet-dry tropics of Australia. We collected blood samples from male and female water pythons held in the laboratory without food and water for 4 weeks. We also collected blood from free-ranging snakes throughout the Austral dry-season. We evaluated plasma osmolality and innate immune function (agglutination, lysis, and bacterial-killing ability) and found that increased osmolality, whether manipulated in the laboratory or as a result of natural water limitation, resulted in enhanced aspects of innate immune performance. Counter-intuitively, snakes in the wild became more hydrated as the dry season progressed, suggesting the dehydrated snakes move to water sources periodically to rehydrate. Comparing our data with those from previous studies, we suspect species divergence in the level of dehydration (i.e., hyperosmolality) that triggers enhanced immune capabilities., (© 2019 Wiley Periodicals, Inc.)

Objective: To examine psychotropic medication prescription practices in federal Canadian penitentiaries., Method: 468 files were drawn from a purposive sample of thirteen Canadian federal institutions representing the five regions, different security levels, and male and female designated facilities. Information on the names of all psychotropic medications prescribed, indications for use, dosage, frequency, and route of administration was retrieved. Designation of approved or off-label use of medications was determined by consulting: (1) the Health Canada (2016) Drug Product Database, (2) the Canadian Compendium of Pharmaceuticals and Specialties 2016, and (3) the American Hospital Formulary Service Drug Information 2016. Prescription rates were examined by gender, Indigenous ancestry, drug class, institutional infractions, and current offence., Results: 36.2% of prescriptions for psychotropic medication were coded as 'off-label'. Anxiolytic/hypnotics drugs were the psychotropic drugs most commonly used for off-label purposes. There were no differences in the prevalence of approved versus off-label prescriptions based on Indigenous ancestry or gender, and no pattern of elevated off-label prescription practices for offenders involved in institutional misconducts or those sentenced for the most serious crimes., Conclusions: The rates of prescribing 'off-label' psychotropic medication are not elevated relative to other correctional settings or to rates cited in Canadian surveys conducted in the community.

We detected a disease syndrome in free-ranging Australian cane toads involving atypical behavior and emaciation that is associated with a previously undescribed Entamoeba sp. that infiltrates the colonic lining, causing it to slough. The organism may become seasonally pathogenic when toads are under hydric and nutritional stress.

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., (© 2018 The Authors. Journal of Animal Ecology © 2018 British Ecological Society.)

Parasites can enhance their fitness by modifying the behavior of their hosts in ways that increase rates of production and transmission of parasite larvae. We used an antihelminthic drug to experimentally alter infections of lungworms ( Rhabdias pseudosphaerocephala ) in cane toads ( Rhinella marina ). We then compared subsequent behaviors of dewormed toads versus toads that retained infections. Both in the laboratory and in the field, the presence of parasites induced hosts to select higher body temperatures (thereby increasing rates of lungworm egg production), to defecate in moister sites, and to produce feces with higher moisture content (thereby enhancing survival of larvae shed in feces). Because those behavioral modifications enhance rather than decrease parasite fitness, they are likely to have arisen as adaptive manipulations of host behavior rather than as host adaptations to combat infection or as nonadaptive consequences of infection on host physiology. However, the mechanisms by which lungworms alter cane toad thermal preference and defecation are not known. Although many examples of host manipulation by parasites involve intermediate hosts facilitating their own demise, our findings indicate that manipulation of definitive hosts can be as subtle as when and where to defecate.

Biological invasions can stimulate rapid shifts in organismal performance, via both plasticity and adaptation. We can distinguish between these two proximate mechanisms by rearing offspring from populations under identical conditions and measuring their locomotor abilities in standardized trials. We collected adult cane toads ( Rhinella marina ) from invasive populations that inhabit regions of Australia with different climatic conditions. We bred those toads and raised their offspring under common-garden conditions before testing their locomotor performance. At high (but not low) temperatures, offspring of individuals from a hotter location (northwestern Australia) outperformed offspring of conspecifics from a cooler location (northeastern Australia). This disparity indicates that, within less than 100 years, thermal performance in cane toads has adapted to the novel abiotic challenges that cane toads have encountered during their invasion of tropical Australia.

We incubated the eggs of field-caught keelback snakes (Tropidonophis mairii) on wet versus dry substrates to explore impacts of incubation conditions on white blood cell (WBC) concentrations and differential WBC counts of hatchlings. In a second, independent study young snakes were released into the field, allowing us to explore fitness correlates of WBC profiles. Dry incubation reduced embryonic survival and hatchling body size, thus decoupling egg size from hatchling size. Incubation conditions also altered WBC profiles. Lymphocyte and azurophil counts were related to hatchling body size but not to initial egg mass, whereas heterophil counts were related to both of these traits. The egg-size effect on heterophil counts may reflect a maternal effect on offspring immune configuration. The ratio of heterophils to lymphocytes (an index of stress) was higher in hatchlings from eggs incubated on dry substrates. Snakes with higher counts of lymphocytes at hatching were more likely to be recaptured as adults (H:L; an index of survival), whereas snakes with higher basophil counts exhibited more rapid growth. In summary, our experiments show that incubation moisture levels influence the immune configuration of hatchling snakes, and that variation among individuals in WBC counts at hatching is a significant predictor of an individual's fitness after it is released into the wild. The demonstrated link between incubation conditions and offspring fitness is likely to impose strong selection on maternal nest-site choice., (© 2018 Wiley Periodicals, Inc.)

The cane toad ( Rhinella marina ) has undergone rapid evolution during its invasion of tropical Australia. Toads from invasion front populations (in Western Australia) have been reported to exhibit a stronger baseline phagocytic immune response than do conspecifics from range core populations (in Queensland). To explore this difference, we injected wild-caught toads from both areas with the experimental antigen lipopolysaccharide (LPS, to mimic bacterial infection) and measured whole-blood phagocytosis. Because the hypothalamic-pituitary-adrenal axis is stimulated by infection (and may influence immune responses), we measured glucocorticoid response through urinary corticosterone levels. Relative to injection of a control (phosphate-buffered saline), LPS injection increased both phagocytosis and the proportion of neutrophils in the blood. However, responses were similar in toads from both populations. This null result may reflect the ubiquity of bacterial risks across the toad's invaded range; utilization of this immune pathway may not have altered during the process of invasion. LPS injection also induced a reduction in urinary corticosterone levels, perhaps as a result of chronic stress., Competing Interests: Lee A. Rollins is an Academic Editor for PeerJ.

The immunological and pathological consequences of parasite infection can be more rigorously assessed from experimental manipulation than from correlational studies of natural infections. We used anthelmintic treatment to experimentally decrease intensities of lungworm infection in captive and free-ranging wild cane toads to assess parasite impacts on host immune responses. First, we administered the anthelmintic drug Ivermectin to both infected and uninfected toads, to distinguish drug effects per se from the impacts of killing lungworms. Worms began dying and decomposing <48 h after injection. The only immunological variables that were affected by anthelmintic treatment were bactericidal capacity of the blood which increased in parasitized toads (presumably triggered by decomposing worms in the lungs), and the phagocytic capacity of blood (which increased in both infected and uninfected toads); the latter effect presumably was caused by the injection of Ivermectin per se rather than removal of parasites. Second, we looked at correlates of variation in the infection intensity induced by de-worming (in both captive and free-ranging toads) over an eight-week period. Heavier lungworm infection was associated with increased phagocytic ability of the host's blood, and a reduction in the host's liver mass (and hence, energy stores). Experimental de-worming thus revealed pathological and immunological costs of the presence of lungworms, and of their removal by anthelmintic injection.

The challenge of identifying the proximate causes and ecological consequences of phenotypic variation can be facilitated by studying traits that are usually but not always bilaterally symmetrical; deviations from symmetry likely reflect disrupted embryogenesis. Based on a 19-year mark-recapture study of >1300 slatey-grey snakes (Stegonotus cucullatus) in tropical Australia, and incubation of >700 eggs, we document developmental and ecological correlates of two morphological traits: asymmetry and fragmentation of head scales. Asymmetry was directional (more scales on the left side) and was higher in individuals with lower heterozygosity, but was not heritable. In contrast, fragmentation was heritable and was higher in females than males. Both scale asymmetry and fragmentation were increased by rapid embryogenesis but were not affected by hydric conditions during incubation. Snakes with asymmetry and fragmentation exhibited slightly lower survival and increased (sex-specific) movements, and females with more scale fragmentation produced smaller eggs. Counterintuitively, snakes with more asymmetry had higher growth rates (possibly reflecting trade-offs with other traits), and snakes with more fragmentation had fewer parasites (possibly due to lower feeding rates). Our data paint an unusually detailed picture of the complex genetic and environmental factors that, by disrupting early embryonic development, generate variations in morphology that have detectable correlations with ecological performance.

Colonial-nesting organisms can strongly alter the chemical and biotic conditions around their aggregation sites, with cascading impacts on other components of the ecosystem. In tropical Australia, Metallic Starlings (Aplonis metallica) nest in large colonies far above the forest canopy, in emergent trees. The ground beneath those trees is open, in stark contrast to the dense foliage all around. We surveyed the areas beneath 27 colony trees (and nearby randomly chosen trees lacking bird colonies) to quantify the birds' impacts on soil and vegetation characteristics, and to test alternative hypotheses about the proximate mechanisms responsible for the lack of live vegetation beneath colony trees. Nutrient levels were greatly elevated beneath colony trees (especially, those with larger colonies), potentially reaching levels toxic to older trees. However, seedlings thrived in the soil from beneath colony trees. The primary mechanism generating open areas beneath colony trees is disturbance by scavengers (feral pigs and native Turkeys) that are attracted in vast numbers to these nutrient hotspots. Seedlings flourished within exclosures inaccessible to vertebrate herbivores, but were rapidly consumed if unprotected. Our results contrast with previous studies of colonies of seabirds on remote islands, where a lack of large terrestrial herbivores results in bird colonies encouraging rather than eliminating vegetation in areas close to the nesting site. In our continental study system, scavengers may rapidly dilute the spatial heterogeneity generated by the massive nutrient subsidy from bird colonies., (© 2016 by the Ecological Society of America.)

Invasive species often exhibit rapid evolutionary changes, and can provide powerful insights into the selective forces shaping phenotypic traits that influence dispersal rates and/or sexual interactions. Invasions also may modify sexual dimorphism. We measured relative lengths of forelimbs and hindlimbs of more than 3000 field-caught adult cane toads ( Rhinella marina ) from 67 sites in Hawai'i and Australia (1-80 years post-colonization), along with 489 captive-bred individuals from multiple Australian sites raised in a 'common garden' (to examine heritability and reduce environmental influences on morphology). As cane toads spread from east to west across Australia, the ancestral condition (long limbs, especially in males) was modified. Limb length relative to body size was first reduced (perhaps owing to natural selection on locomotor ability), but then increased again (perhaps owing to spatial sorting) in the invasion vanguard. In contrast, the sex disparity in relative limb length has progressively decreased during the toads' Australian invasion. Offspring reared in a common environment exhibited similar geographical divergences in morphology as did wild-caught animals, suggesting a genetic basis to the changes. Limb dimensions showed significant heritability (2-17%), consistent with the possibility of an evolved response. Cane toad populations thus have undergone a major shift in sexual dimorphism in relative limb lengths during their brief (81 years) spread through tropical Australia.

Police agencies in Canada and elsewhere have received much criticism over how they respond to persons with serious mental disorders. The adequacy of training provided to police officers on mental health issues and in particular on recognizing indicators of serious mental disorders has been a major concern. This paper describes the process that led to the development of a new brief mental health screener (interRAI Brief Mental Health Screener, BMHS) designed to assist police officers to better identify persons with serious mental disorders. The interRAI BMHS was developed in collaboration with interRAI, an international, not-for-profit consortium of researchers. The government of Ontario had previously partnered with interRAI to develop and implement the Resident Assessment Instrument for Mental Health (RAI-MH), the assessment system mandated for use on all persons admitted into inpatient psychiatric care in the province. Core items on the interRAI BMHS were obtained through analysis (N=41,019) of RAI-MH data together with input from representatives from health care, police services, and patient groups. Two police services in southwestern Ontario completed forms (N=235) on persons thought to have a mental disorder. Patient records were later accessed to determine patient disposition. The use of summary and inferential statistics revealed that the variables significantly associated with being taken to hospital by police included performing a self-injurious act in the past 30days, and others being concerned over the person's risk for self-injury. Variables significantly associated with being admitted included abnormal thought process, delusions, and hallucinations. The results of the study indicate that the 14-variable algorithm used to construct the interRAI BMHS is a good predictor of who was most likely to be taken to hospital by police officers and who was most likely to be admitted. The instrument is an effective means of capturing and standardizing police officer observations enabling them to provide more and better quality information to emergency department (ED) staff. Teaching police officers to use the form constitutes enhanced training on major indicators of serious mental disorders. Further, given that items on the interRAI BMHS are written in the language of the health system, language acts as common currency between police officers and ED staff laying the foundation for a more collaborative approach between the systems., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Although widespread declines in anuran populations have attracted considerable concern, the stochastic demographics of these animals make it difficult to detect consistent trends against a background of spatial and temporal variation. To identify long-term trends, we need datasets gathered over long time periods, especially from tropical areas where anuran biodiversity is highest. We conducted road surveys of four anurans in the Australian wet-dry tropics on 4637 nights over a 16-year period. Our surveys spanned the arrival of invasive cane toads (Rhinella marina), allowing us to assess the invader's impact on native anuran populations. Our counts demonstrate abrupt and asynchronous shifts in abundance and species composition from one year to the next, not clearly linked to rainfall patterns. Typically, periods of decline in numbers of a species were limited to 1-2 years and were followed by 1- to 2-year periods of increase. No taxa showed consistent declines over time, although trajectories for some species showed significant perturbations coincident with the arrival of toads. None of the four focal frog species was less common at the end of the study than at the beginning, and three of the species reached peak abundances after toad arrival. Survey counts of cane toads increased rapidly during the initial stage of invasion but have subsequently declined and fluctuated. Distinguishing consistent declines versus stochastic fluctuations in anuran populations requires extensive time-series analysis, coupled with an understanding of the shifts expected under local climatic conditions. This is especially pertinent when assessing impacts of specific perturbations such as invasive species.

Since 2006, 3 new disease syndromes have emerged in farmed saltwater crocodiles (Crocodylus porosus) in the Northern Territory of Australia. We describe the syndromes through a retrospective study of laboratory findings from 187 diagnostic cases submitted to Berrimah Veterinary Laboratories between 2005 and 2014. The first syndrome was characterized by conjunctivitis and/or pharyngitis (CP), primarily in hatchlings. Herpesviruses were isolated in primary crocodile cell culture, or were detected by PCR directly from conjunctiva or pharyngeal tissue, in 21 of 39 cases of CP (54%), compared with 9 of 64 crocodiles without the syndrome (14%, p < 0.0001). Chlamydiaceae were detected by PCR in conjunctiva or pharyngeal tissue of 55% of 29 CP cases tested, and of these, 81% also contained herpesvirus. The second syndrome occurred in juveniles and growers exhibiting poor growth, and was characterized histologically by systemic lymphoid proliferation and nonsuppurative encephalitis (SLPE). Herpesviruses were isolated or detected by PCR from at least 1 internal organ in 31 of 33 SLPE cases (94%) compared with 5 of 95 crocodiles without the syndrome (5%, p < 0.0001). The third syndrome, characterized by multifocal lymphohistiocytic infiltration of the dermis (LNS), occurred in 6 harvest-sized crocodiles. Herpesviruses were isolated from at least 1 skin lesion in 4 of these 6 cases. Although our study revealed strong associations between herpesvirus and the CP and SLPE syndromes, the precise nature of the role of herpesvirus, along with the pathogenesis and epidemiology of the syndromes, requires further investigation., (© 2016 The Author(s).)

Like most ectothermic vertebrates, keelback snakes (Tropidonophis mairii) do not exhibit parental care. Thus, offspring must possess an immune system capable of dealing with challenges such as pathogens, without assistance from an attendant parent. We know very little about immune system characteristics of neonatal reptiles, including the magnitude of heritability and other maternal influences. To identify sources of variation in circulating white blood cell (WBC) concentrations and differentials, we examined blood smears from 246 hatchling snakes and their field-caught mothers. WBC concentrations were lower in hatchlings than in adults, and hatchlings had more basophils and fewer azurophils than adults. A hatchling keelback's WBC differential was also influenced by its sex and body size. Although hatchling WBC measures exhibited negligible heritability, they were strongly influenced by maternal body size and parasite infection (but not by maternal body condition, relative clutch mass or time in captivity). Larger mothers produced offspring with more azurophils and fewer lymphocytes. The mechanisms and consequences of WBC variation are currently unknown, but if these maternal effects enhance offspring fitness, the impact of maternal body size on reproductive success may be greater than expected simply from allometric increases in the numbers and sizes of progeny.

There have been numerous empirical and agent-based modelling studies on the spatial self-structuring of traits, particularly in regard to dispersal ability (termed spatial sorting) of cane toads in northern Australia, but few mathematical modelling studies. In this study, we formulate a reaction-diffusion based partial-integro-differential equation model based on an earlier model by Bouin et al. (2012) to examine this spatial self-structuring of traits in both a cane toad population and lungworm parasite population, which evolves with the cane toad population. In particular, the traits we focus on are dispersal ability for the cane toad population and both prepatent period and larval size for the lungworm parasite population. Apart from the spatial self-structuring of these traits, our results confirm a number of observations made in empirical and agent-based studies; particularly, that there is a noticeable lag between the host and parasite population which is critically dependent on the parasite functional response to host densities, that older populations regress back to lower dispersal speeds and that spatial sorting can still occur with a disadvantage in reproductivity and/or survival in more motile individuals. Moreover, we find that such a disadvantage in reproductivity and/or survival is unlikely to be large if spatial sorting is to have a noticeable effect on the rate of range expansion, as it has been observed to have over the last 60 years in northern Australia., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Many invading species have brought devastating parasites and diseases to their new homes, thereby imperiling native taxa. Potentially, though, invaders might have the opposite effect. If they take up parasites that otherwise would infect native taxa, but those parasites fail to develop in the invader, the introduced species might reduce parasite burdens of the native fauna. Similarly, earlier exposure to the other taxon's parasites might 'prime' an anuran's immune system such that it is then able to reject subsequent infection by its own parasite species. Field surveys suggest that lungworm counts in native Australian frogs decrease after the arrival of invasive cane toads (Rhinella marina), and laboratory studies confirm that native lungworm larvae enter, but do not survive in, the toads. In laboratory trials, we confirmed that the presence of anurans (either frogs or toads) in an experimental arena reduced uptake rates of lungworm larvae by anurans that were later added to the same arena. However, experimental exposure to lungworms from native frogs did not enhance a toad's ability to reject subsequent infection by its own lungworm species.

Biological invasions can bring both the invader and native taxa into contact with novel parasites. As cane toads ( Rhinella marina ) have spread through Australia, they have encountered lungworms (Rhabdias hylae) that occur in native frogs. Field surveys suggest that these lungworms have not host-switched to toads. In our laboratory studies, R. hylae infected cane toads as readily as it infected native frogs, but failed to reach the lungs of the novel host (i.e., were killed by the toads' immune response). Plausibly, then, R. hylae might reduce the viability both of their native hosts (frogs, that can exhibit high parasite burdens) and cane toads (that must deal with infective larvae traveling through the host body). Our laboratory trials suggest, however, that the impacts of the parasite on infected anuran hosts (both frogs and toads) were minimal, with no significant decrements to host survival, activity, growth, or locomotor performance. Ironically, the lack of impact of the parasite on its native hosts appears to be an outcome of co-evolution (frogs tolerate the lungworm), whereas the lack of impact on the novel host is due to a lack of co-evolution (toads can recognize and eliminate the lungworm).

Background: Individual movement is critical to organismal fitness and also influences broader population processes such as demographic stochasticity and gene flow. Climatic change and habitat fragmentation render the drivers of individual movement especially critical to understand. Rates of movement of free-ranging animals through the landscape are influenced both by intrinsic attributes of an organism (e.g., size, body condition, age), and by external forces (e.g., weather, predation risk). Statistical modelling can clarify the relative importance of those processes, because externally-imposed pressures should generate synchronous displacements among individuals within a population, whereas intrinsic factors should generate consistency through time within each individual. External and intrinsic factors may vary in importance at different time scales., Results: In this study we focused on daily displacement of an ambush-foraging snake from tropical Australia (the Northern Death Adder Acanthophis praelongus), based on a radiotelemetric study. We used a mixture of spectral representation and Bayesian inference to study synchrony in snake displacement by phase shift analysis. We further studied autocorrelation in fluctuations of displacement distances as "one over f noise". Displacement distances were positively autocorrelated with all considered noise colour parameters estimated as >0. We show how the methodology can reveal time scales of particular interest for synchrony and found that for the analysed data, synchrony was only present at time scales above approximately three weeks., Conclusion: We conclude that the spectral representation combined with Bayesian inference is a promising approach for analysis of movement data. Applying the framework to telemetry data of A. praelongus, we were able to identify a cut-off time scale above which we found support for synchrony, thus revealing a time scale where global external drivers have a larger impact on the movement behaviour. Our results suggest that for the considered study period, movement at shorter time scales was primarily driven by factors at the individual level; daily fluctuations in weather conditions had little effect on snake movement.

The cane toad invasion in Australia provides a robust opportunity to clarify the infection process in co-evolved versus de novo host-parasite interactions. We investigated these infection dynamics through histological examination following experimental infections of metamorphs of native frogs (Cyclorana australis) and cane toads (Rhinella marina) with Rhabdias hylae (the lungworm found in native frogs) and Rhabdias pseudosphaerocephala (the lungworm found in cane toads). Cane toads reared under continuous exposure to infective larvae of the frog lungworm were examined after periods of 2, 6, 10 and 15 days. Additionally, both toads and frogs were exposed for 24 h to larvae of either the toad or the frog lungworm, and examined 2, 5, 10 and 20 days post-treatment. R. hylae (frog) lungworms entered cane toads and migrated through the body but were not found in the target tissue, the lungs. Larvae of both lungworm species induced inflammation in both types of hosts, although the immune response (relative numbers of different cell types) differed between hosts and between parasite species. Co-evolution has modified the immune response elicited by infection and (perhaps for that reason) has enhanced the parasite's ability to survive and to reach the host's lungs.