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Concept: Toads


The introduced cane toad (Bufo marinus) poses a major threat to biodiversity due to its lifelong toxicity. Several terrestrial native Australian vertebrates are adapting to the cane toad’s presence and lab trials have demonstrated that repeated exposure to B. marinus can result in learnt avoidance behaviour. Here we investigated whether aversion learning is occurring in aquatic ecosystems by comparing cane toad naïve and sympatric populations of crimson spotted rainbow fish (Melanotaenia duboulayi). The first experiment indicated that fish from the sympatric population had pre-existing aversion to attacking cane toad tadpoles but also showed reduced attacks on native tadpoles. The second experiment revealed that fish from both naïve and sympatric populations learned to avoid cane toad tadpoles following repeated, direct exposure. Allopatric fish also developed a general aversion to tadpoles. The aversion learning abilities of both groups was examined using an experiment involving novel distasteful prey items. While both populations developed a general avoidance of edible pellets in the presence of distasteful pellets, only the sympatric population significantly reduced the number of attacks on the novel distasteful prey item. These results indicate that experience with toxic prey items over multiple generations can enhance avoidance leaning capabilities via natural selection.

Concepts: Ecology, Learning, Allopatric speciation, Toads, Bufo, Cane toad


Phenotypic differences among species may evolve through genetic accommodation, but mechanisms accounting for this process are poorly understood. Here we compare hormonal variation underlying differences in the timing of metamorphosis among three spadefoot toads with different larval periods and responsiveness to pond drying. We find that, in response to pond drying, Pelobates cultripes and Spea multiplicata accelerate metamorphosis, increase standard metabolic rate (SMR), and elevate whole-body content of thyroid hormone (the primary morphogen controlling metamorphosis) and corticosterone (a stress hormone acting synergistically with thyroid hormone to accelerate metamorphosis). In contrast, Scaphiopus couchii has the shortest larval period, highest whole-body thyroid hormone and corticosterone content, and highest SMR, and these trait values are least affected by pond drying among the three species. Our findings support that the atypically rapid and canalized development of S. couchii evolved by genetic accommodation of endocrine pathways controlling metamorphosis, showing how phenotypic plasticity within species may evolve into trait variation among species.

Concepts: Evolution, Metabolism, Endocrinology, Thyroid, Thyroid hormone, Adrenal cortex, Amphibian, Toads


The Coatzacoalcos Region in Veracruz, Mexico houses one of the most important industrial complexes in Mexico and Latin America. Lead is an ubiquitous environmental pollutant which represents a great risk to human health and ecosystems. Amphibian populations have been recognized as biomonitors of changes in environmental conditions. The purpose of this research is to measure exposure to lead and evaluate hematological and biochemical effects in specimens of giant toads (Rhinella marina) taken from three areas surrounding an industrial complex in the Coatzacoalcos River downstream. Lead levels in toads' blood are between 10.8 and 70.6 μg/dL and are significantly higher in industrial sites. We have found a significant decrease in the delta-aminolevulinic acid dehydratase (δ-ALAD) activity in blood from 35.3 to 78 % for the urban-industrial and industrial sites, respectively. In addition, we have identified a strong inverse relationship between the δ-ALAD activity and the blood lead levels (r = -0.84, p < 0.001). Hemoglobin and mean corpuscular hemoglobin levels, as well as the condition factor, are found to be lower at industrial sites compared with the reference sites. Our results suggest that the R. marina can be considered a good biomonitor of the δ-ALAD activity inhibition and hematological alterations at low lead concentrations.

Concepts: Protein, Red blood cell, PH, Amphibian, Toad, Mexico City, Lead poisoning, Toads


In biological invasions, rates of range expansion tend to accelerate through time. What kind of benefits to more rapidly dispersing organisms might impose natural selection for faster rates of dispersal, and hence the evolution of range-edge acceleration? We can answer that question by comparing fitness-relevant ecological traits of individuals at the invasion front compared with conspecifics in the same area a few years post-invasion. In tropical Australia, the rate of invasion by cane toads (Rhinella marina) has increased substantially over recent decades, due to shifts in heritable traits. Our data on field-collected cane toads at a recently invaded site in the Australian wet-dry tropics span a 5-year period beginning with toad arrival. Compared with conspecifics that we monitored in the same sites post-invasion, toads in the invasion vanguard exhibited higher feeding rates, larger energy stores, better body condition and faster growth. Three processes may have contributed to this pattern: (i) higher prey availability at the front (perhaps due to reduced competition from conspecifics); (ii) the lack of viability-reducing parasites and pathogens in invasion-front toads; and (iii) distinctive (active, fast-growing) phenotypes of the invasion-front toads. Nutritional benefits to individuals in the invasion vanguard (whether because of higher prey availability, or lower pathogen levels) thus may have conferred a selective advantage to accelerated dispersal in this system.

Concepts: Natural selection, Evolution, Biology, Acceleration, Invasion, Toads, Invasions, Dwayne Johnson


Non-invasive measurement of urinary corticosterone and testosterone metabolites in amphibians provides opportunities for endocrine studies of responses to physiological and psychological stressors. Typically, corticosterone metabolite concentrations increase in frog urine within 1-2h of a mild capture and handling stress protocol. However, no study has investigated the effect of duration of manual restraint on the changes in corticosterone and reproductive hormones in amphibians. We quantified urinary corticosterone and testosterone metabolite responses for 8h following various durations of manual restraint (control, 5, 15 or 30min) in adult male cane toads (Rhinella marina) under controlled laboratory conditions. All toads had a corticosterone stress response over 8h to our standard capture and handling stressor. The mean corticosterone stress response was significantly higher after 15 or 30min restraint in comparison to the control (no restraint) or to 5min restraint. Manual restraint for 5, 15 or 30min caused a significant reduction in urinary testosterone concentrations over 8h. We also provide a novel method of quantifying plasticity in corticosterone stress responses in amphibians with respect to restraint duration using the concept of a “reaction norm”. The reaction norm, which was calculated as slope of the regression line of integrated corticosterone response against restraint duration, was 9.69 (pg corticosterone/μg creatinineh)/min for male toads. In summary, corticosterone and testosterone responses to restraint are affected by restraint duration in male toads. Glucocorticoid reaction norms can be applied to study the change in physiological stress hormonal response with respect to restraint duration in other amphibian species.

Concepts: Reproduction, Hormone, Adrenal cortex, Amphibian, Frog, Toad, Toads, True toad


Objective-To evaluate the righting reflex after topical application of a sevoflurane jelly in cane toads (Bufo marinus). Animals-8 cane toads. Procedures-Toads were 6 to 8 months of age and weighed (mean ± SD) 142.0 ± 25.2 g. Sevoflurane jelly was applied to the dorsum of each toad at a dose of 25 μL/g in trial 1 and 37.5 μL/g in trial 2. Toads were placed in dorsal recumbency every 30 seconds until loss of the righting reflex. Jelly was then removed by rinsing the toads with tap water. Toads were then left undisturbed in dorsal recumbency until return of the righting reflex. Chamber sevoflurane concentration was measured to determine vaporization. Results-6 of 8 toads in trial 1 and 8 of 8 toads in trial 2 lost the righting reflex. Mean ± SD time to loss of the reflex was 8.2 ± 1.3 minutes for trial 1 and 8.3 ± 0.9 minutes for trial 2; this difference was not significant. Mean ± SD time to return of the reflex was 25.6 ± 26.2 minutes for trial 1 and 84.4 ± 47.2 minutes for trial 2; this difference was significant. Chamber sevoflurane concentration did not change significantly, compared with baseline (time 0) concentration, at any time in trial 1; however, there was a significant change in chamber sevoflurane concentration from baseline (time 0) concentration in trial 2. Chamber sevoflurane concentrations were not significantly different between trial 1 and trial 2 at any time. Mean ± SD chamber sevoflurane concentration was 0.46 ± 0.2% for trial 1 and 0.57 ± 0.28% for trial 2. Conclusions and Clinical Relevance-Sevoflurane jelly applied topically at a dose of 37.5 μL/g induced a more reliable loss of righting reflex and longer recovery time than when applied at a dose of 25 μL/g in cane toads.

Concepts: Topical, Lidocaine, Toads, Cane toad


If invasive species use chemical weapons to suppress the viability of conspecifics, we may be able to exploit those species-specific chemical cues for selective control of the invader. Cane toads (Rhinella marina) are spreading through tropical Australia, with negative effects on native species. The tadpoles of cane toads eliminate intraspecific competitors by locating and consuming newly laid eggs. Our laboratory trials show that tadpoles find those eggs by searching for the powerful bufadienolide toxins (especially, bufogenins) that toads use to deter predators. Using those toxins as bait, funnel-traps placed in natural waterbodies achieved near-complete eradication of cane toad tadpoles with minimal collateral damage (because most native (non-target) species are repelled by the toads' toxins). More generally, communication systems that have evolved for intraspecific conflict provide novel opportunities for invasive-species control.

Concepts: Ecology, English-language films, Amphibian, Frog, Introduced species, Toads, True toad, Psychoactive toad


Biological invasions can induce rapid evolutionary change. As cane toads (Rhinella marina) have spread across tropical Australia over an 80-year period, their rate of invasion has increased from around 15 to 60 km per annum. Toads at the invasion front disperse much faster and further than conspecifics from range-core areas, and their offspring inherit that rapid dispersal rate. We investigated morphological changes that have accompanied this dramatic acceleration, by conducting three-dimensional morphometric analyses of toads from both range-core and invasion-front populations. Morphology of heads, limbs, pectoral girdles and pelvic girdles differed significantly between toads from the two areas, ranging from 0.5% to 16.5% difference in mean bone dimensions between populations, with invasion-front toads exhibiting wider forelimbs, narrower hindlimbs and more compact skulls. Those changes plausibly reflect an increased reliance on bounding (multiple short hops in quick succession) rather than separate large leaps. Within an 80-year period, invasive cane toads have converted the basic anuran body plan - which evolved for occasional large leaps to evade predators - into a morphotype better-suited to sustained long-distance travel.

Concepts: Evolution, Biology, Amphibian, Frog, Toad, Toads, True toad, Dwayne Johnson


At the edge of a biological invasion, evolutionary processes (spatial sorting, natural selection) often drive increases in dispersal. Although numerous traits influence an individual’s displacement (e.g. speed, stamina), one of the most important is path straightness. A straight (i.e. highly correlated) path strongly enhances overall dispersal rate relative to time and energetic cost. Thus, we predict that, if path straightness has a genetic basis, organisms in the invasion vanguard will exhibit straighter paths than those following behind. Our studies on invasive cane toads (Rhinella marina) in tropical Australia clearly support this prediction. Radio-tracking of field-collected toads at a single site showed that path straightness steadily decreased over the first 10 years post-invasion. Consistent with an evolved (genetic) basis to that behavioural shift, path straightness of toads reared under common garden conditions varied according to the location of their parents' origin. Offspring produced by toads from the invasion vanguard followed straighter paths than did those produced by parents from long-established populations. At the individual level, offspring exhibited similar path straightness to their parents. The dramatic acceleration of the cane toad invasion through tropical Australia has been driven, in part, by the evolution of a behavioural tendency towards dispersing in a straight line.

Concepts: Natural selection, Evolution, Biology, Curve, Line, Analytic geometry, Toads, Line segment


Management of invasive vertebrates is a crucial component of conservation. Trapping reproductive adults is often effective for control, and modification of traps may greatly increase their attractiveness to such individuals. Cane toads (Rhinella marina) are invasive, and males use advertisement vocalisations to attract reproductive females. In amphibians, including toads, specific structural parameters of calls (e.g., dominant frequency and pulse rate) may be attractive to females. Some cane toad traps use an artificial advertisement vocalisation to attract toads. We determined whether variation of the call’s parameters (volume, dominant frequency, and pulse rate) could increase the capture rate of gravid females.

Concepts: Human, Male, Reproduction, Female, Amphibian, Toad, Toads, True toad