Elevated corticosterone levels are associated with increased immunocompetence in male toads, both when calling and under experimental conditions.

Circulating androgens can influence immune responses and sexual traits in male vertebrates. However, in the last 20 years, glucocorticoids have also been implicated as a possible source of variation in male sexual traits and immunocompetence. In this context, we studied the relations between male vocal mating display, immunity, androgens, and glucocorticoids in the explosive breeding toad Rhinella granulosa. In the field, males with high calling effort display either high- or low-corticosterone (CORT) plasma levels, but only males with both high calling effort and high CORT plasma levels showed high bacterial killing ability (BKA), suggesting that the acute CORT elevation can be immunostimulatory. CORT treatments increased BKA in laboratory experiments, confirming the functional relationship observed in the field. However, toads treated with a low dose of CORT increased BKA for 10 h after the treatment, while toads that received a high dose increased BKA for only 1 h after the treatment. These results indicate that different CORT doses can result in temporal differences in the immune response. We did not find any relationship between calling effort, immune response (BKA and PHA swelling response), and testosterone plasma levels in the field, or any effects of testosterone treatment on immunocompetence. Our results suggest a complex relationship between calling effort and immunity, mediated by CORT plasma levels.

Influence of temperature, size and confinement on testosterone and corticosterone levels in breeding male spined toads (Bufo spinosus).

Winter breeding amphibians opportunistically engage in reproductive behaviour when environmental conditions become favourable. In such explosive breeding systems, males strongly compete for reproductive females. Although most research has been oriented on species in which males use mating calls to attract females, many high-density explosive breeding amphibians do not rely on mating calls. In such systems, larger and stronger males are thought to have significant advantages to access reproductive females. Testosterone (T) is expected to increase with the physical attributes that facilitate access to females, while increased corticosterone levels (CORT) are needed to sustain the energetic requirements associated with mating behaviour. In this study, we investigated how environmental temperature, and body size influence testosterone and corticosterone in males of an explosive winter-breeding species with low investment in mating call, the spined toad (Bufo spinosus). We found that both baseline CORT and T were positively correlated with environmental temperature. Interestingly, despite a remarkable range of variation in CORT and T, there was no evidence that either hormone was correlated with body size. Finally, we found no effect of confinement (13 h of captivity) on circulating CORT and T levels. This suggests that breeding male toads may be relatively insensitive to stress in order to maintain reproductive effort during their short mating period. Future studies should investigate both the influence of the phenology of breeding and the social interactions on these hormonal levels.

Breeding biology and the evolution of dynamic sexual dichromatism in frogs.

Dynamic sexual dichromatism is a temporary colour change between the sexes and has evolved independently in a wide range of anurans, many of which are explosive breeders wherein males physically compete for access to females. Behavioural studies in a few species indicate that dynamic dichromatism functions as a visual signal in large breeding aggregations; however, the prevalence of this trait and the social and environmental factors underlying its expression are poorly understood. We compiled a database of 178 anurans with dynamic dichromatism that include representatives from 15 families and subfamilies. Dynamic dichromatism is common in two of the three subfamilies of hylid treefrogs. Phylogenetic comparative analyses of 355 hylid species (of which 95 display dynamic dichromatism) reveal high transition rates between dynamic dichromatism, ontogenetic (permanent) dichromatism and monochromatism reflecting the high evolutionary lability of this trait. Correlated evolution in hylids between dynamic dichromatism and forming large breeding aggregations indicates that the evolution of large breeding aggregations precedes the evolution of dynamic dichromatism. Multivariate phylogenetic logistic regression recovers the interaction between biogeographic distribution and forming breeding aggregations as a significant predictor of dynamic dichromatism in hylids. Accounting for macroecological differences between temperate and tropical regions, such as seasonality and the availability of breeding sites, may improve our understanding of ecological contexts in which dynamic dichromatism is likely to arise in tropical lineages and why it is retained in some temperate species and lost in others.

Does repeated human handling of study animals during the mating season affect their offspring?

Minimizing disturbance of study animals is a major consideration in ethological and ecological research design. One nearly universal type of disturbance is the handling of study animals as a component of trial setup. Even low to moderate levels of handling can be a substantial stressor to study animals, which may negatively affect their offspring via maternal effects. Understanding how routine human handling and manipulation may affect the outcome of research studies is therefore critical for interpreting study outcomes. We tested whether repeatedly handling and manipulating (i.e., manually disengaging) amplexed pairs of wood frogs (Rana sylvatica [Lithobates sylvaticus]), which have an explosive breeding season, would affect their reproductive output and offspring fitness. Handling and manipulation did not alter any parameter that we measured: reproductive timing, hatching success, and offspring larval duration, survival, and size at metamorphosis. These results suggest that handling and manipulation by researchers may have a negligible effect on wood frog reproduction and offspring fitness. It is possible that many species that are commonly used in reproductive studies because they suppress behavioral and physiological responses during the mating season are likewise unaffected by human handling. Nevertheless, researchers should examine possible consequences of methodological interventions on their study species in order to determine any potential influence on their results. Having a broad understanding of these effects on species that have robust or dampened stress responsiveness during the breeding season would be useful for making generalizations about potential effects.

Dynamic sexual dichromatism produces a sex signal in an explosively breeding Neotropical toad: A model presentation experiment.

Many animals breed in large mating aggregations, where males must rapidly discriminate between prospective mates and rivals. Selection may favour features that facilitate rapid discrimination in these aggregations. The explosively breeding Neotropical Yellow Toad, Incilius luetkenii, exhibits a rapid and dramatic colour change; males change from a cryptic brown to a conspicuous lemon yellow for their brief breeding period. Females, in contrast, remain cryptic brown throughout the year. The function of this temporary, sex-specific colour change is unknown. We tested the hypothesis that yellow colouration displayed by male I. luetkenii facilitates sex recognition during both daytime and nighttime mating aggregations. We created yellow and brown model toads and presented them to males during a breeding event. Male I. luetkenii responded significantly more intensely to brown models compared to yellow models, approaching them and making more amplexus attempts on the brown versus yellow models. This strong pattern held true regardless of ambient light intensity, making this the first study to expose a dynamic colour signal that operates during both day and night. Our results indicate that male I. luetkenii use colouration to quickly discriminate between males and females during their brief, explosive mating aggregations. Our findings suggest that the rapid, dramatic colour change of male I. luetkenii facilitates sex recognition, which could provide a significant fitness advantage to males in the form of reduced energy expenditure and reduced risk of injury by other males. Dynamic dichromatism may provide similar fitness benefits in any organisms that mate in large, competitive aggregations.