Trained quantity discrimination in invasive red-eared slider and a comparison with the native stripe-necked turtle.

Little is known about the behavioral and cognitive traits that best predict invasion success. Evidence is mounting that cognitive performance correlates with survival and fecundity, two pivotal factors for the successful establishment of invasive populations. We assessed the quantity discrimination ability of the globally invasive red-eared slider (Trachemys scripta elegans). We further compared it to that of the native stripe-necked turtle (Mauremys sinensis), which has been previously evaluated for its superior quantity discrimination ability. Specifically, our experimental designs aimed to quantify the learning ability as numerosity pairs increased in difficulty (termed fixed numerosity tests), and the immediate response when turtles were presented with varied challenges concurrently in the same tests (termed mixed numerosity tests). Our findings reaffirm the remarkable ability of freshwater turtles to discern numerical differences as close as 9 vs 10 (ratio = 0.9), which was comparable to the stripe-necked turtle's performance. However, the red-eared slider exhibited a moderate decrease in performance in high ratio tests, indicating a potentially enhanced cognitive capacity to adapt to novel challenges. Our experimental design is repeatable and is adaptable to a range of freshwater turtles. These findings emphasize the potential importance of cognitive research to the underlying mechanisms of successful species invasions.

Captivity induces large and population-dependent brain transcriptomic changes in wild-caught cane toads (Rhinella marina).

Gene expression levels are key molecular phenotypes at the interplay between genotype and environment. Mounting evidence suggests that short-term changes in environmental conditions, such as those encountered in captivity, can substantially affect gene expression levels. Yet, the exact magnitude of this effect, how general it is, and whether it results in parallel changes across populations are not well understood. Here, we take advantage of the well-studied cane toad, Rhinella marina, to examine the effect of short-term captivity on brain gene expression levels, and determine whether effects of captivity differ between long-colonized and vanguard populations of the cane toad's Australian invasion range. We compared the transcriptomes of wild-caught toads immediately assayed with those from toads captured from the same populations but maintained in captivity for seven months. We found large differences in gene expression levels between captive and wild-caught toads from the same population, with an over-representation of processes related to behaviour and the response to stress. Captivity had a much larger effect on both gene expression levels and gene expression variability in toads from vanguard populations compared to toads from long-colonized areas, potentially indicating an increased plasticity in toads at the leading edge of the invasion. Overall, our findings indicate that short-term captivity can induce large and population-specific transcriptomic changes, which has significant implications for studies comparing phenotypic traits of wild-caught organisms from different populations that have been held in captivity.

A new protocol for investigating visual two-choice discrimination learning in lizards.

One of the most widely studied abilities in lizards is discrimination learning. The protocols used to test lizards are often novel or adapted from other taxa without proper validation. We need to ensure that tests of discrimination learning are appropriate and properly applied in lizards so that robust inferences can be made about cognitive ability. Here, we present a new protocol for testing lizard discrimination learning that incorporates a target training procedure, uses many daily trials for efficiency and reinforcement, and has a robust, validated, learning criterion. We trained lizards to touch a cue card using operant conditioning and tested lizards separately on a colour, and pattern discrimination test. Lizards successfully learnt to touch a cue card and to discriminate between light and dark blue but had issues discriminating the patterns. After modifying the test procedure, some lizards reached criterion, revealing possible issues with stimulus processing and interference of generalisation. Here, we describe a protocol for operant conditioning and two-choice discrimination learning in lizards with a robust learning criterion that can help researcher better design future studies on discrimination learning in lizards.

A Smaller Habenula is Associated with Increasing Intensity of Sexual Selection.

The habenula is a small structure in the brain that acts as a relay station for neural information, helping to modulate behaviour in response to variable and unpredictable stimuli. Broadly, it is evolutionarily conserved in structure and connectivity across vertebrates and is the most prominent bilaterally asymmetric structure in the brain. Nonetheless, comparative evolutionary studies of the habenula are virtually non-existent. Here, we examine the volumes of the medial and lateral habenular subregions, in both hemispheres, across a group of Australian agamid lizards in the genus Ctenophorus. In males, we found bilaterally asymmetrical selection on the lateral habenula to become smaller with increasing intensity of sexual selection, possibly as a mechanism to increase aggressive responses. In females, we found bilaterally symmetrical selection on both the medial and lateral subregions to become smaller with increasing sexual selection. This is consistent with sexual selection increasing motivation to reproduce and the habenula's well-characterized role in controlling and modifying responses to rewarding stimuli. However, as there are currently no studies addressing habenular function in reptiles, it is difficult to draw more precise conclusions. As has happened recently in biomedical neuroscience, it is time for the habenula to receive greater attention in evolutionary neuroscience.

Sperm cryopreservation in an Australian skink (Eulamprus quoyii).

Assisted reproductive technologies for population and genetic management for threatened herpetofauna have grown substantially in the past decade. Here we describe experiments to optimise sperm cryopreservation in a model squamate, the eastern water skink Eulamprus quoyii . Small, concentrated volumes of highly motile spermatozoa were reliably collected from adult male E. quoyii by non-lethal ventral massage. Samples were used to: (1) test whether protein-rich diluents, namely Beltsville poultry semen extender (BPSE) and TES and Tris (TEST) yolk buffer (TYB), improve post-thaw quality metrics compared with Dulbecco's phosphate-buffered saline (DPBS); and (2) compare the efficacy of these diluents in combination with either 1.35M glycerol or 1.35M dimethyl sulfoxide (DMSO) at two freezing rates, fast (approximately -20°C min-1 ) versus slow (-6°C min-1 ). Glycerol and DMSO performed equally well in preserving spermatozoa under slow freezing rates. Under these conditions, the use of the complex diluents BPSE and TYB significantly improved post-thaw total motility compared with DPBS. Complex interactions occurred between cryodiluent type, cryoprotectant and freezing rate when testing fast versus slow freezing rates among treatment groups. Under slow freezing rates, DMSO was better at preserving membrane integrity and motility, regardless of diluent type, but successful fast freezing required complex diluents to support motility and membrane integrity, which has implications for implementation in a field setting.

Investment in chemical signalling glands facilitates the evolution of sociality in lizards.

The evolution of sociality and traits that correlate with, or predict, sociality, have been the focus of considerable recent study. In order to reduce the social conflict that ultimately comes with group living, and foster social tolerance, individuals need reliable information about group members and potential rivals. Chemical signals are one such source of information and are widely used in many animal taxa, including lizards. Here, we take a phylogenetic comparative approach to test the hypothesis that social grouping correlates with investment in chemical signalling. We used the presence of epidermal glands as a proxy of chemical investment and considered social grouping as the occurrence of social groups containing both adults and juveniles. Based on a dataset of 911 lizard species, our models strongly supported correlated evolution between social grouping and chemical signalling glands. The rate of transition towards social grouping from a background of 'epidermal glands present' was an order of a magnitude higher than from a background of 'no epidermal glands'. Our results highlight the potential importance of chemical signalling during the evolution of sociality and the need for more focused studies on the role of chemical communication in facilitating information transfer about individual and group identity, and ameliorating social conflict.

A deepening understanding of animal culture suggests lessons for conservation.

A key goal of conservation is to protect biodiversity by supporting the long-term persistence of viable, natural populations of wild species. Conservation practice has long been guided by genetic, ecological and demographic indicators of risk. Emerging evidence of animal culture across diverse taxa and its role as a driver of evolutionary diversification, population structure and demographic processes may be essential for augmenting these conventional conservation approaches and decision-making. Animal culture was the focus of a ground-breaking resolution under the Convention on the Conservation of Migratory Species of Wild Animals (CMS), an international treaty operating under the UN Environment Programme. Here, we synthesize existing evidence to demonstrate how social learning and animal culture interact with processes important to conservation management. Specifically, we explore how social learning might influence population viability and be an important resource in response to anthropogenic change, and provide examples of how it can result in phenotypically distinct units with different, socially learnt behavioural strategies. While identifying culture and social learning can be challenging, indirect identification and parsimonious inferences may be informative. Finally, we identify relevant methodologies and provide a framework for viewing behavioural data through a cultural lens which might provide new insights for conservation management.

Sexual selection on performance traits in an Australian lizard with alternative reproductive tactics.

Sexual selection shapes the adaptive landscape in complex ways that lead to trait integration. Much of our understanding of selection comes from studies of morphological traits. However, few studies attempt to quantify the form and direction of selection on performance even though it is predicted to be a more direct target of selection in nature. We measured sexual selection on performance traits (bite force, sprint speed and endurance) in an Australian lizard, the Eastern water skink (Eulamprus quoyii). We first staged 123 contests between size-matched males to investigate whether performance traits were important in determining contest outcome. In a second experiment, we established six breeding populations in large replicate semi-natural enclosures to estimate whether performance traits predicted reproductive success. Our results show that none of the performance measures were important in predicting contest outcome and were not generally strong predictors of reproductive success. However, our analyses suggest a complex fitness landscape driven by males adopting different alternative reproductive tactics (ARTs). We provide a rare test of the role performance plays in sexual selection and highlight the need to test common assumptions regarding the link between maximal performance and fitness. Our results suggest that performance traits may not necessarily be direct targets of sexual selection, but rather indirect targets through their integration with morphological and/or behavioural traits, highlighting a need for more explicit tests of the predicted links between performance and fitness.

Superior continuous quantity discrimination in a freshwater turtle.

BACKGROUND: Quantity discrimination, the ability to discriminate a magnitude of difference or discrete numerical information, plays a key role in animal behavior. While quantitative ability has been well documented in fishes, birds, mammals, and even in previously unstudied invertebrates and amphibians, it is still poorly understood in reptiles and has never been tested in an aquatic turtle despite the fact that evidence is accumulating that reptiles possess cognitive skills and learning ability. To help address this deficiency in reptiles, we investigated the quantitative ability of an Asian freshwater turtle, Mauremys sinensis, using red cubes on a white background in a trained quantity discrimination task. While spontaneous quantity discrimination methods are thought to be more ecologically relevant, training animals on a quantity discrimination task allows more comparability across taxa. RESULTS: We assessed the turtles' quantitative performance in a series of tests with increasing quantity ratios and numerosities. Surprisingly, the turtles were able to discriminate quantities of up to 9 versus 10 (ratio = 0.9), which shows a good quantitative ability that is comparable to some endotherms. Our results showed that the turtles' quantitative performance followed Weber's law, in which success rate decreased with increasing quantity ratio across a wide range of numerosities. Furthermore, the gradual improvement of their success rate across different experiments and phases suggested that the turtles possess learning ability. CONCLUSIONS: Reptile quantitative ability has long been ignored and therefore is likely under-estimated. More comparative research on numerical cognition across a diversity of species will greatly contribute to a clearer understanding of quantitative ability in animals and whether it has evolved convergently in diverse taxa.

Convergent evolution of skin surface microarchitecture and increased skin hydrophobicity in semi-aquatic anole lizards.

Animals that habitually cross the boundary between water and land face specific challenges with respect to locomotion, respiration, insulation, fouling and waterproofing. Many semi-aquatic invertebrates and plants have developed complex surface microstructures with water-repellent properties to overcome these problems, but equivalent adaptations of the skin have not been reported for vertebrates that encounter similar environmental challenges. Here, we document the first evidence of evolutionary convergence of hydrophobic structured skin in a group of semi-aquatic tetrapods. We show that the skin surface of semi-aquatic species of Anolis lizards is characterized by a more elaborate microstructural architecture (i.e. longer spines and spinules) and a lower wettability relative to closely related terrestrial species. In addition, phylogenetic comparative models reveal repeated independent evolution of enhanced skin hydrophobicity associated with the transition to a semi-aquatic lifestyle, providing evidence of adaptation. Our findings invite a new and exciting line of inquiry into the ecological significance, evolutionary origin and developmental basis of hydrophobic skin surfaces in semi-aquatic lizards, which is essential for understanding why and how the observed skin adaptations evolved in some and not other semi-aquatic tetrapod lineages.

Do I stay or do I go? Shifts in perch use by lizards during morning twilight suggest anticipatory behaviour.

Anticipatory behaviour is the expectation of a near-future event based on information processed in the past and influences an animal's tactical decisions, particularly when there are significant fitness consequences. The grass lizard (Takydromus viridipunctatus) perches on blades of grass at night which likely reduces the probability of predation by terrestrial predators such as snakes, rodents and shrews. During twilight (starting 30 min before sunrise), they move from above the grass to within grass clumps and this is thought to afford the lizard protection while reducing detection by avian predators. Here, we examined how lizards shift their behaviour as a function of visual detectability to their primary predator, the cattle egret (Bubulcus ibis). We show that the lizards shift from their perch site during twilight at the earliest time at which egrets depart communal roosts. At the same time, visual modelling shows a dramatic increase in the detectability of the lizards to the visual system of egrets. Therefore, anticipatory behaviour in response to environmental cues acts to reduce predation risk as lizards become more conspicuous and predators become more active. Grass lizard anticipatory behaviour appears to be finely tuned by natural selection to adjust to temporal changes in predation risk.

Sperm Storage in a Family-Living Lizard, the Tree Skink (Egernia striolata).

The ability to produce viable offspring without recently mating, either through sperm storage or parthenogenesis, can provide fitness advantages under a suite of challenging ecological scenarios. Using genetic analysis, we demonstrate that 3 wild-caught female Tree Skinks (Egernia striolata) reproduced in captivity with no access to males for over a year, and that this is best explained by sperm storage. To the best of our knowledge, this is the first time female sperm storage has been documented in any monogamous family-living reptile, including social Australian egerniine skinks (from the subfamily Egerniinae). Furthermore, by using paternal reconstruction of genotypes we show that captive-born offspring produced by the same females in the preceding year, presumably without sperm storage, were sired by different males. We qualitatively compared aspects of these females' mates and offspring between years. The parents of each litter were unrelated, but paternal and offspring genotypes from litters resulting from stored sperm were more heterozygous than those inferred to be from recent matings. Family-living egerniine skinks generally have low rates of multiple paternity, yet our study suggests that female sperm storage, potentially from outside social partners, offers the real possibility of benefits. Possible benefits include increasing genetic compatibility of mates and avoiding inbreeding depression via cryptic female choice. Sperm storage in Tree Skinks, a family-living lizard with a monogamous mating system, suggests that females may bet-hedge through extra-pair copulation with more heterozygous males, reinforcing the idea that females could have more control on reproductive outcomes than previously thought.

Context-specific response inhibition and differential impact of a learning bias in a lizard.

Response inhibition (inhibiting prepotent responses) is needed for reaching a more favourable goal in situations where reacting automatically would be detrimental. Inhibiting prepotent responses to resist the temptation of a stimulus in certain situations, such as a novel food item, can directly affect an animal's survival. In humans and dogs, response inhibition varies between contexts and between individuals. We used two contextually different experiments to investigate response inhibition in the eastern water skink (Eulamprus quoyii): reversal of a visual two-choice discrimination and a cylinder detour task. During the two-choice task, half of our lizards were able to reach an initial learning criterion, but, thereafter, did not show consistent performance. Only two individuals reached a more stringent criterion, but subsequently failed during reversals. Furthermore, half of our animals were not able to inhibit a pre-existing side preference which affected their ability to learn during the two-choice task. Skinks were, however, able to achieve a detour around a cylinder performing at levels comparable to brown lemurs, marmosets, and some parrot species. A comparison between the tasks showed that reaching the initial criterion was associated with low success during the detour task, indicating that response inhibition could be context-specific in the water skink. To the best of our knowledge, this is the first study to examine inhibitory control and motor self-regulation in a lizard species.

Testing domain general learning in an Australian lizard.

A key question in cognition is whether animals that are proficient in a specific cognitive domain (domain specific hypothesis), such as spatial learning, are also proficient in other domains (domain general hypothesis) or whether there is a trade-off. Studies testing among these hypotheses are biased towards mammals and birds. To understand constraints on the evolution of cognition more generally, we need broader taxonomic and phylogenetic coverage. We used Australian eastern water skinks (Eulamprus quoyii) with known spatial learning ability in three additional tasks: an instrumental and two discrimination tasks. Under domain specific learning we predicted that lizards that were good at spatial learning would perform less well in the discrimination tasks. Conversely, we predicted that lizards that did not meet our criterion for spatial learning would likewise perform better in discrimination tasks. Lizards with domain general learning should perform approximately equally well (or poorly) in these tasks. Lizards classified as spatial learners performed no differently to non-spatial learners in both the instrumental and discrimination learning tasks. Nevertheless, lizards were proficient in all tasks. Our results reveal two patterns: domain general learning in spatial learners and domain specific learning in non-spatial learners. We suggest that delineating learning into domain general and domain specific may be overly simplistic and we need to instead focus on individual variation in learning ability, which ultimately, is likely to play a key role in fitness. These results, in combination with previously published work on this species, suggests that this species has behavioral flexibility because they are competent across multiple cognitive domains and are capable of reversal learning.

Learning from others: an invasive lizard uses social information from both conspecifics and heterospecifics.

Species that are able to solve novel problems through social learning from either a conspecific or a heterospecific may gain a significant advantage in new environments. We tested the ability of a highly successful invasive species, the Italian wall lizard Podarcis sicula, to solve a novel foraging task when social information was available from both a conspecific and an unfamiliar heterospecific (Podarcis bocagei). We found that Italian wall lizards that had access to social information made fewer errors, regardless of whether the demonstrator was a conspecific or a heterospecific, compared to Italian wall lizards that individually learnt the same task. We suggest that social learning could be a previously underappreciated, advantageous mechanism facilitating invasions.

Archipelagos of the Anthropocene: rapid and extensive differentiation of native terrestrial vertebrates in a single metropolis.

Some of the best evidence for rapid evolutionary change comes from studies of archipelagos and oceanic islands. City parks are analogous systems as they create geographically isolated green spaces that differ in size, structure and complexity. Very little, however, is known about whether city parks within a single urban centre drive selection and result in the diversification of native species. Here, we provide evidence for the rapid genetic and morphological differentiation of a native lizard (Intellagama lesueurii) at four geographically close yet unconnected parks within one city. Year of establishment of each city park varied from 1855 (oldest) to 2001 (youngest) equating to a generation time range of 32 to three generations. Genetic divergence among city park populations was large despite the small pairwise geographic distances (<5 km) and found to be two to three times higher for microsatellites and three to 33 times higher for mtDNA relative to nonurban populations. Patterns of morphological differentiation were also found to be most extensive among the four city park populations. In contrast to nonurban populations, city park populations showed significant differentiation in relative body size, relative head and limb morphology and relative forelimb and hindlimb length. Crucially, we show that these patterns of differentiation are unlikely to have been caused by founder events and/or drift alone. Our results suggest that city park 'archipelagos' could represent theatres for rapid evolution that may, in time, favour adaptive diversification.

Dominance and social information use in a lizard.

There is mounting evidence that social learning is not just restricted to group-living animals, but also occurs in species with a wide range of social systems. However, we still have a poor understanding of the factors driving individual differences in social information use. We investigated the effects of relative dominance on social information use in the eastern water skink (Eulamprus quoyii), a species with age-dependent social learning. We used staged contests to establish dominant-subordinate relationships in pairs of lizards and tested whether observers use social information to more quickly solve both an association and reversal learning task in situations where the demonstrator was either dominant or subordinate. Surprisingly, we found no evidence of social information use, irrespective of relative dominance between observer and demonstrator. However, dominant lizards learnt at a faster rate than subordinate lizards in the associative learning task, although there were no significant differences in the reversal task. In light of previous work in this species, we suggest that age may be a more important driver of social information use because demonstrators and observers in our study were closely size-matched and were likely to be of similar age.

Does social environment influence learning ability in a family-living lizard?

Early developmental environment can have profound effects on individual physiology, behaviour, and learning. In birds and mammals, social isolation during development is known to negatively affect learning ability; yet in other taxa, like reptiles, the effect of social isolation during development on learning ability is unknown. We investigated how social environment affects learning ability in the family-living tree skink (Egernia striolata). We hypothesized that early social environment shapes cognitive development in skinks and predicted that skinks raised in social isolation would have reduced learning ability compared to skinks raised socially. Offspring were separated at birth into two rearing treatments: (1) raised alone or (2) in a pair. After 1 year, we quantified spatial learning ability of skinks in these rearing treatments (N = 14 solitary, 14 social). We found no effect of rearing treatment on learning ability. The number of skinks to successfully learn the task, the number of trials taken to learn the task, the latency to perform the task, and the number of errors in each trial did not differ between isolated and socially reared skinks. Our results were unexpected, yet the facultative nature of this species' social system may result in a reduced effect of social isolation on behaviour when compared to species with obligate sociality. Overall, our findings do not provide evidence that social environment affects development of spatial learning ability in this family-living lizard.

The loneliness of the long-distance toad: invasion history and social attraction in cane toads (Rhinella marina).

Individuals at the leading edge of a biological invasion constantly encounter novel environments. These pioneers may benefit from increased social attraction, because low population densities reduce competition and risks of pathogen transfer, and increase benefits of information transfer. In standardized trials, cane toads (Rhinella marina) from invasion-front populations approached conspecifics more often, and spent more time close to them, than did conspecifics from high-density, long-colonized populations.