Widespread learned predator recognition to an alien predator across populations in an amphibian species.

Alien predators are a major cause of decline and extinction of species worldwide, since native organisms are rarely equipped with specific antipredatory strategies to cope with them. However, phenotypic plasticity and learned predator recognition may help prey populations to survive novel predators. Here we examine geographical variation in the learning ability of larval spadefoot toads (Pelobates cultripes) to recognize invasive predatory crayfish (Procambarus clarkii). We compare the learning-mediated behavioural responses of tadpoles from six populations across two regions in Spain (central and southern), with different histories of exposure to the presence of the invasive species. Two of the populations showed innate recognition of chemical cues from the invasive crayfish, whereas three of them learned to recognize such cues as a threat after conditioning with conspecific alarm cues. Learning abilities did not differ among southern populations, but they did among central populations. We assessed patterns of genetic variation within and among these two regions through microsatellite markers and found low genetic divergence among the southern populations but greater differentiation among the central ones. We hypothesize that similar responses to the invasive crayfish in southern populations may have arisen from a combination of extended historical exposure to this introduced predator (~ 50 y) and higher levels of gene flow, as they inhabit a highly interconnected pond network. In contrast, populations from central Spain show lower connectivity, have been exposed to the invasive crayfish for a shorter period of time, and are more divergent in their plastic responses.

Background matching through fast and reversible melanin-based pigmentation plasticity in tadpoles comes with morphological and antioxidant changes.

Facultative colour change is widespread in the animal kingdom, and has been documented in many distantly related amphibians. However, experimental data testing the extent of facultative colour change, and associated physiological and morphological implications are comparatively scarce. Background matching in the face of spatial and temporal environmental variation is thought to be an important proximate function of colour change in aquatic amphibian larvae. This is particularly relevant for species with long larval periods such as the western spadefoot toad, Pelobates cultripes, whose tadpoles spend up to six months developing in temporary waterbodies with temporally variable vegetation. By rearing tadpoles on different coloured backgrounds, we show that P. cultripes larvae can regulate pigmentation to track fine-grained differences in background brightness, but not hue or saturation. We found that colour change is rapid, reversible, and primarily achieved through changes in the quantity of eumelanin in the skin. We show that this increased eumelanin production and/or maintenance is also correlated with changes in morphology and oxidative stress, with more pigmented tadpoles growing larger tail fins and having an improved redox status.

Assessment of predation risk through conspecific cues by anuran larvae.

Accurate assessment of predation risk is critical for prey survival during predator-prey interactions. Prey can assess predation risk by the presence of cues dropped by predators themselves, but they can also gather information about risk level through cues released by other prey, avoiding the hazard of being in close proximity to predators. In this study, we examine the ability of anuran larvae (Pelobates cultripes) to detect predation risk indirectly when they are in contact with conspecifics that have been recently exposed to chemical stimuli from natural predators (larvae of aquatic beetles). In a first experiment, we confirmed that larvae exposed to predator cues exhibited innate defensive behavior, indicating that they perceived the risk of predation and, thus, could potentially act as risk indicators for naïve conspecifics. In a second experiment, we observed that unexposed larvae paired with a startled conspecific adjusted their antipredator behavior, presumably by mirroring conspecifics' behavior and/or using chemical cues from their partners as a risk information source. This cognitive ability of tadpoles to assess predation risk through conspecific cues might play an important role in their interaction with predators, facilitating the early detection of potential threats to elicit appropriate antipredator responses and increase the chances of survival.

Pigmentation plasticity enhances crypsis in larval newts: associated metabolic cost and background choice behaviour.

In heterogeneous environments, the capacity for colour change can be a valuable adaptation enhancing crypsis against predators. Alternatively, organisms might achieve concealment by evolving preferences for backgrounds that match their visual traits, thus avoiding the costs of plasticity. Here we examined the degree of plasticity in pigmentation of newt larvae (Lissotriton boscai) in relation to predation risk. Furthermore, we tested for associated metabolic costs and pigmentation-dependent background choice behaviour. Newt larvae expressed substantial changes in pigmentation so that light, high-reflecting environment induced depigmentation whereas dark, low-reflecting environment induced pigmentation in just three days of exposure. Induced pigmentation was completely reversible upon switching microhabitats. Predator cues, however, did not enhance cryptic phenotypes, suggesting that environmental albedo induces changes in pigmentation improving concealment regardless of the perceived predation risk. Metabolic rate was higher in heavily pigmented individuals from dark environments, indicating a high energetic requirement of pigmentation that could impose a constraint to larval camouflage in dim habitats. Finally, we found partial evidence for larvae selecting backgrounds matching their induced phenotypes. However, in the presence of predator cues, larvae increased the time spent in light environments, which may reflect a escape response towards shallow waters rather than an attempt at increasing crypsis.

Low levels of chemical anthropogenic pollution may threaten amphibians by impairing predator recognition.

Recent studies suggest that direct mortality and physiological effects caused by pollutants are major contributing factors to global amphibian decline. However, even sublethal concentrations of pollutants could be harmful if they combined with other factors to cause high mortality in amphibians. Here we show that sublethal concentrations of pollutants can disrupt the ability of amphibian larvae to recognize predators, hence increasing their risk of predation. This effect is indeed much more important since very low amounts of pollutants are ubiquitous, and environmental efforts are mostly directed towards preventing lethal spills. We analyzed the effects of two common contaminants (humic acid and ammonium nitrate) on the ability of tadpoles of the western spadefoot toad (Pelobates cultripes) to recognize chemical cues from a common predator, nymphs of the dragonfly Anax imperator. We compared the swimming activity of tadpoles in the presence and absence of water-borne chemical cues from dragonflies at different concentrations of humic acid and ammonium nitrate. Tadpoles reduced swimming activity in response to predator cues in the absence of pollutants, whereas they remained unresponsive to these cues when either humic acid or ammonium nitrate was added to the water, even at low concentrations. Moreover, changes in tadpole activity associated with the pollutants themselves were non-significant, indicating no toxic effect. Alteration of the natural chemical environment of aquatic systems by pollutants may be an important contributing cause for declines in amphibian populations, even at sublethal concentrations.

Asymmetry in food handling behavior of a tree-dwelling rodent (Sciurus vulgaris).

Asymmetry in motor patterns is present in a wide variety of animals. Many lateralized behaviors seem to depend on brain asymmetry, as it is the case of different tasks associated to food handling by several bird and mammal species. Here, we analyzed asymmetry in handling behavior of pine cones by red squirrels (Sciurus vulgaris). Red squirrels devote most of their daily activity to feeding, thus this species constitutes an appropriate model for studying asymmetry in food processing. We aimed to explore 1) the potential lateralization in handling of pine cones by squirrels, 2) the dominant pattern for this behavior (left- vs. right-handed), and 3) whether this pattern varies among populations and depending on the pine tree species available. Results revealed that red squirrels handle pine cones in an asymmetrical way, and that direction of asymmetry varies among populations and seems to be determined more by local influences rather than by the pine tree species.

Honest sexual signaling in turtles: experimental evidence of a trade-off between immune response and coloration in red-eared sliders Trachemys scripta elegans.

Sexual signals can be evolutionarily stable if they are honest and condition dependent or costly to the signaler. One possible cost is the existence of a trade-off between maintaining the immune system and the elaboration of ornaments. This hypothesis has been experimentally tested in some groups of animals but not in others such as turtles. We experimentally challenged the immune system of female red-eared sliders Trachemys scripta elegans, with a bacterial antigen (lipopolysaccharide (LPS)) without pathogenic effects to explore whether the immune activation affected visual colorful ornaments of the head. The LPS injection altered the reflectance patterns of color ornaments. In comparison to the control animals, the yellow chin stripes of injected animals exhibited (1) reduced brightness, (2) lower long wavelength (>470 nm) reflectance, and (3) lower values for carotenoid chroma. The postorbital patches of injected individuals also showed reduced very long wavelength (>570 nm) reflectance but did not change in carotenoid chroma. Thus, experimental turtles showed darker and less "yellowish" chin stripes and less "reddish" postorbital patches at the end of the experiment, whereas control turtles did not change their coloration. This is the first experimental evidence supporting the existence of a trade-off between the immune system and the expression of visual ornaments in turtles. We suggest that this trade-off may allow turtles to honestly signal individual quality via characteristics of coloration, which may have an important role in intersexual selection processes.

Effects of body temperature on righting performance of native and invasive freshwater turtles: consequences for competition.

Righting behavior of aquatic turtles might be subject to coadaptation pressures between preferred basking temperature and locomotion, given that it is mainly performed on land and may critically determine the survival of turtles. We analyzed the effect of body temperature (T(b)) on righting performance of two species of freshwater turtles, the endangered native Spanish terrapin (Mauremys leprosa), and the red-eared slider (Trachemys scripta elegans), an introduced invasive species that is displacing native turtles in the Iberian Peninsula. Interspecific differences in morphology, basking requirements and behavioral responses have been found between Spanish terrapins and introduced sliders. Therefore, we hypothesized that T(b) might differentially affect righting behavior of these two turtle species. We found a clear effect of T(b) on righting response of both M. leprosa and T. scripta, with the performance enhanced at the preferred basking temperature of each turtle species. These results suggest that righting might be coadapted to preferred basking temperature in freshwater turtles. Also, M. leprosa required longer times to right on average than T. scripta, which denotes a higher efficiency of introduced sliders at righting performance. These interspecific behavioral asymmetries in righting performance between native and exotic turtles might contribute to the greater competitive ability of introduced T. scripta, favoring the expansion of exotic sliders in the new environments in which they are introduced, in detriment to native Spanish terrapins.

Feeding status and basking requirements of freshwater turtles in an invasion context.

Thermoregulatory behavior and feeding status are strongly related in ectotherms. A trade-off between maintenance of energy balance and digestion efficiency has been recently proposed to affect thermoregulation in these animals. On the other hand, competition for basking sites has been described between Iberian turtles and the introduced red-eared slider (Trachemys scripta elegans). T. scripta negatively interferes with basking behavior of native turtles and benefits from a greater capacity to retain body heat, which may likely result in thermoregulatory advantages for the introduced sliders. Consequently, complex effects and alterations in metabolic rates of native turtles might derive from a deficient basking behavior. We compared the basking requirements of the endangered native Spanish terrapin (Mauremys leprosa) and those of the introduced red-eared slider, analyzing the upper set point temperature (USP) (defined as the body temperature at which basking ceased) of both native and introduced turtles, under feeding and fasting conditions. We found higher values of USP in the native species, and a reduction of this temperature associated with food deprivation in the two turtle species. This adjustment of thermoregulatory behavior to the nutritional status found in freshwater turtles suggests that ectotherms benefit from metabolic depression as an adaptive mechanism to preserve energy during periods of fasting. However, a reduction in metabolic rates induced by competition with sliders might lead M. leprosa to a prolonged deficiency of their physiological functions, thus incurring increased predation risk and health costs, and ultimately favoring the recession of this native species in Mediterranean habitats.