Exploring Sound Emission of the Lizard Pristidactylus valeriae.

Lizards, except geckos, are generally considered voiceless organisms, although some species emit oral sounds. For most of these "vocal lizards", however, there is almost no information on the characteristics of the sounds, precluding exploration of the functionality and evolution of the sounds. Pristidactylus are known as "grunter lizards" since individuals emit oral sounds under predation risk. We explored the characteristics of the sounds emitted by P. valeriae, recording 17 adults and 1 juvenile when they were threatened and captured by a predator. Only adults emitted sounds with open mouths and displayed aggressive postures, e.g., biting attempts. These sounds correspond to hisses, which lack amplitude or frequency modulation. The lizards emitted longer hisses when threatened than when captured by the predator, which may provide honest information on individuals' ability to escape. In addition, males may experience higher distress during threats since their hisses had higher aggregate entropy than those of the females. Finally, hissing has been documented in four of the five Leiosauridae genera, the family to which Pristidactylus belongs, suggesting that sound emission is ancestral to the family.

Response of the weeping lizard to distress calls: the effect of witnessing predation.

Escaping from predation saves life, but costs energy and time. The "threat-sensitive predator-avoidance" hypothesis proposes that prey may modulate their antipredator responses, and thus the associated costs, in accordance with the magnitude of predation risk. This process requires that prey accurately assess this risk by decoding available information from various sources. For example, distress calls are uttered by prey when a predator traps them and can serve as public information on predation risk. Such is the case for the weeping lizard whose distress calls trigger immobility in conspecifics. Here, we tested whether this antipredator response of the weeping lizard is modulated by witnessing predation. We exposed lizards to distress calls alone or paired with models of a prey (conspecific), a predator (snake), or a predatory event (a snake subjugating the conspecific). Data show that the sole presence of the predator or prey paired with distress calls seems not to modulate the antipredator responses. Contrarily, witnessing a predatory event associated with calls intensified antipredator responses; lizards reduced their activity for longer and avoided proximity to the stimuli, which may decrease predation risk by reducing the likelihood of being detected by the predator. We conclude that the weeping lizard can use multisensorial public information to assess predation risk and modulate its antipredator responses.

Geographic variation in the matching between call characteristics and tympanic sensitivity in the Weeping lizard.

Effective communication requires a match among signal characteristics, environmental conditions, and receptor tuning and decoding. The degree of matching, however, can vary, among others due to different selective pressures affecting the communication components. For evolutionary novelties, strong selective pressures are likely to act upon the signal and receptor to promote a tight match among them. We test this prediction by exploring the coupling between the acoustic signals and auditory sensitivity in Liolaemus chiliensis, the Weeping lizard, the only one of more than 285 Liolaemus species that vocalizes. Individuals emit distress calls that convey information of predation risk to conspecifics, which may respond with antipredator behaviors upon hearing calls. Specifically, we explored the match between spectral characteristics of the distress calls and the tympanic sensitivities of two populations separated by more than 700 km, for which previous data suggested variation in their distress calls. We found that populations differed in signal and receptor characteristics and that this signal variation was explained by population differences in body size. No precise match occurred between the communication components studied, and populations differed in the degree of such correspondence. We suggest that this difference in matching between populations relates to evolutionary processes affecting the Weeping lizard distress calls.

Lizard predation by spiders: A review from the Neotropical and Andean regions.

Vertebrate predation by invertebrates has been classically underexplored and thus underestimated, despite the fact that many arthropods consume vertebrates. To shed some light on the relevance that spider predation may have upon lizards in the Neotropical and Andean regions, we compiled the available information in the literature on this trophic interaction. We found 50 reports of spiders consuming lizards in these regions, and the 88% of these were from the Neotropical region. Spiders belong to eight families, but Ctenidae and Theraphosidae were the most frequently reported predators. Lizards belong to 12 families, and the most commonly consumed species corresponded to the families Dactyloidae (all Anolis lizards), Gymnophthalmidae, and Sphaerodactylidae. Data suggest trophic spider-lizard associations between Ctenidae and Dactyloidae, followed by Theraphosidae and Liolaemidae. The body sizes of the spiders and lizards showed a positive relationship, and spiders were smaller than their prey. We conclude that various spider taxa can be considered lizard predators and they may be ecologically important in the Neotropical and Andean regions. However, spiders of prime predation relevance seem to be those of the Ctenidae and Theraphosidae families.

Without a body of evidence and peer review, taxonomic changes in Liolaemidae and Tropiduridae (Squamata) must be rejected.

In his recent self-published book "Reptiles en Chile", Diego Demangel Miranda presented 13 taxonomic changes for liolaemid and tropidurid lizards. While these could be considered validly published according to the International Code of Zoological Nomenclature, we show that these taxonomic propositions lack the necessary scientific rigor in terms of replicability, specimen work, lack of peer review and that they do not follow best practices accepted by the herpetological community. Therefore, we hereby invalidate all 13 taxonomic changes proposed in this book, leaving the taxonomy unaffected. Finally, we call attention to the potentially negative consequences of using these taxonomic changes in conservation and environmental impact studies as incorrect decisions might be taken in relation to the species involved.

The songs of male pied flycatchers: exploring the legacy of the fathers.

Singing is a key element of songbirds' behavioral repertoire, particularly for males, which sing during the breeding season to defend resources against other males and to attract females. Different song traits may convey honest information about males' qualities or conditions, which may be used by females to select their mates. Traits under strong sexual selection have an important component of additive genetic variation (i.e., the main genetic inheritance from parents), and so relatively high heritability; therefore, it can be expected that song traits also do. Although the act of singing is an innate behavior, and thus, genetically determined, songbirds need to learn their songs and therefore the genetic contribution to song traits may be reduced by the effect of environmental factors. We tested this hypothesis in seven song traits recorded in the long-distance migratory bird, the pied flycatcher (Ficedula hypoleuca). From a 23-year database (1992-2015), we obtained songs for 28 father-son pairs, and for each song trait we applied parent-offspring regressions to estimate heritability. The type of syllables sung are learned from tutors, and here we also determined the cultural contribution of fathers to the song repertoires of their sons, by quantifying the percentage of syllables that sons shared with their fathers, and compared this with what sons shared with other males in the population (e.g., neighbors). The heritabilities of song traits were highly variable (ranging from -0.22 to 0.56), but most of these were around zero and none of them were significant. These results indicate that the seven song traits are most likely determined by environmental factors. Sons shared more syllables with their fathers than with neighbors (21% vs. 3%), suggesting that fathers are important song tutors during the nestling period. We conclude that there is a cultural inheritance from fathers to their sons' syllable repertoires, but there is no strong evidence for a genetic contribution of fathers to the seven song traits studied.

Comparing the antipredator behaviour of two sympatric, but not syntopic, Liolaemus lizards.

The microhabitat preferences of prey animals can modulate how they perceive predation risk, and therefore, their antipredator behaviour. We tested under standardized conditions how microhabitat preferences of two Liolaemus lizards affected their responses when confronted with two types of ambush predators (raptor vs. snake), under two levels of predation risk (low vs. high). These lizard species are sympatric, but not syntopic; L. chiliensis basks on bushes, a complex microhabitat that may provide protection against visual predators, while L. nitidus prefers open microhabitats, basking on the top of large bare rocks, highly exposed to visual predators. If microhabitat complexity modulates the antipredator response, L. chiliensis may perceive lower predation risk, exhibiting lower intensity of antipredator responses than L. nitidus. Both species reduced their activity after being exposed to both predators, but lizards differed in the assessment of predation risk; L. nitidus reduced its activity independently of the predation risk experienced, while L. chiliensis only reduced its activity in the high-risk condition. The microhabitat preferences shaped during the evolution of these species seem to modulate their perception of predation risk, which may cause interspecific differences in the associated costs of their antipredator responses.

Advancement of spring arrival in a long-term study of a passerine bird: sex, age and environmental effects.

In migratory birds, mistimed arrival might have negative consequences for individual fitness, causing population declines. This may happen if arrival time is not synchronized with breeding time, especially when earlier springs favour earlier reproduction. We studied spring arrival time to the breeding areas in a pied flycatcher Ficedula hypoleuca population in southern Norway during a 30-year period (1985-2014). We investigated trends in arrival both for the entire population and for different population fractions (e.g. early vs. late arrivals). We also studied sex and age class differences, along with repeatability of arrival. Finally, we explored how arrival is influenced by environmental conditions at the areas birds use throughout the year, using mixed-effects models and quantile regressions with individual-based data. Spring arrival advanced over five days, at a similar rate through the entire population. Males and adult birds arrived earlier than females and yearlings. Arrival was significantly repeatable for males and females. Birds arrived earlier in years with high temperature and rainfall at the breeding grounds, and low NDVI both on the Iberian Peninsula and in central Europe. Later fractions of the population showed a steeper response to these environmental variables. This intra-population heterogeneity in the responses to the environment probably stems from a combination between the different selection pressures individuals are subject to and their age-related experience. Our results highlight the importance of studying how migration phenology is affected by the environment not only on the breeding grounds but also on the other areas birds use throughout the year.

Identification and molecular characterization of five putative toxins from the venom gland of the snake Philodryas chamissonis (Serpentes: Dipsadidae).

Philodryas chamissonis is a rear-fanged snake endemic to Chile. Its bite produces mild to moderate symptoms with proteolytic and anti-coagulant effects. Presently, the composition of the venom, as well as, the biochemical and structural characteristics of its toxins, remains unknown. In this study, we cloned and reported the first full-length sequences of five toxin-encoding genes from the venom gland of this species: Type III snake venom metalloprotease (SVMP), snake venom serine protease (SVSP), Cysteine-rich secretory protein (CRISP), α and ß subunits of C-type lectin-like protein (CLP) and C-type natriuretic peptide (NP). These genes are highly expressed in the venom gland and their sequences exhibited a putative signal peptide, suggesting that these are components of the venom. These putative toxins had different evolutionary relationships with those reported for some front-fanged snakes, being SVMP, SVSP and CRISP of P. chamissonis closely related to the toxins present in Elapidae species, while NP was more related to those of Viperidae species. In addition, analyses suggest that the α and ß subunits of CLP of P. chamissonis might have a α-subunit scaffold in common with Viperidae species, whose highly variable C-terminal region might have allowed the diversification in α and ß subunits. Our results provide the first molecular description of the toxins possibly implicated in the envenomation of prey and humans by the bite of P. chamissonis.

The relation between hairpin formation by mitochondrial WANCY tRNAs and the occurrence of the light strand replication origin in Lepidosauria.

Mitochondrial light strand DNA replication is initiated at light strand replication origins (OLs), short stem-loop hairpins formed by the heavy strand DNA. OL-like secondary structures are also formed by heavy strand DNA templating for the five tRNAs adjacent to OLs, the WANCY tRNA cluster. We tested whether natural OL absence associates with greater capacities for formation of OL-like structures by WANCY tRNA genes. Using lepidosaurian taxa (Sphenodon, lizards and amphisbaenids), we compared WANCY tRNA capacities to form OL-like structures between 248 taxa possessing an OL with 131 taxa without OL (from different families). On average, WANCY tRNA genes form more OL-like structures in the absence of a regular OL than in its presence. Formation of OL-like structures by WANCY tRNAs follows hierarchical patterns that may reduce competition between the tRNA's translational function and its secondary OL function: the rarer the tRNA's cognate amino acid, the greater the capacity to form OL-like structures. High OL-forming capacities for neighboring tRNAs are avoided. Because OL absence usually occurs in taxa with reduced genomes, increased formation of OL-like structures by WANCY tRNAs might result from selection for greater metabolic efficiency. Further analyses suggest that OL loss is one of the latest steps in genome reduction, and promotes the increase in formation of OL-like structures by WANCY tRNA genes in Lepidosauria.

Tetracoding increases with body temperature in Lepidosauria.

Codons expanded by a silent position (quadruplet or tetracodons) may solve the conundrum that at life's origins, the weak tricodon-anticodon interactions could not promote translation in the absence of complex ribosomes. Modern genomes have isolated tetracodons resulting from insertion mutations. Some bioinformatic analyses suggest that tetracoding stretches overlap with regular mitochondrial protein coding genes. These tetragenes are probably decoded by (antisense) tRNAs with expanded anticodons. They are GC-rich, which produce stronger basepairs than A:T interactions, suggesting expression at high temperatures. The hypothesis that tetracoding is an adaptation to high temperatures is tested here by comparing predicted mitochondrial tetracoding in Lepidosauria (lizards, amphisbaenia, and Sphenodon), in relation to body temperature, expecting more tetracoding in species with high body temperature. The association between tRNAs with expanded anticodons and tetracoding previously described for mammals and Drosophila is confirmed for Lepidosauria. Independent evidence indicates that tetracoding increases with body temperature, supporting the hypothesis that tetracoding is an adaptation for efficient translation when conditions (temperature) make triplet codon-anticodons too unstable to allow efficient protein elongation.

Evolution of thermal physiology in Liolaemus lizards: adaptation, phylogenetic inertia, and niche tracking.

Microevolutionary studies often find that complex quantitative characters are highly evolvable and adapted to the local environment, while macroevolutionary studies often show evidence of strong phylogenetic effects and stasis. In this contribution, we show how phylogenetic comparative methods can be used to test hypotheses that may help resolve this paradox. As a test case, we studied the interplay between adaptation and phylogenetic inertia on the thermobiology of 32 species of Liolaemus (Squamata: Liolaemidae), a genus of South American lizards living under diverse climatic conditions. Despite a strong phylogenetic effect in the preferred (selected) body temperature, we found clear evidence that this variable is adapted to local temperature and climate. After controlling for adaptation to the thermal environment, little influence of phylogeny was left. This indicates that the phylogenetic effect was not caused by a lag or slowness in adaptation but primarily by the distribution of the thermal environments on the phylogeny. This can be due to thermal niche tracking. In contrast, we found little or no evidence for adaptation to the thermal environment in either cooling or heating rates, critical thermal minimum, or body size.

Heterogeneity of voltage- and chemosignal-activated response profiles in vomeronasal sensory neurons.

Liolaemus lizards were explored to ascertain whether they would make an amenable model to study single-cell electrophysiology of neurons in the vomeronasal organ (VNO). Despite a rich array of chemosensory-related behaviors chronicled for this genus, no anatomical or functional data exist for the VNO, the organ mediating these types of behaviors. Two Liolaemus species (L. bellii and L. nigroviridis) were collected in Central Chile in the Farellones Mountains and transported to the United States. Lizards were subjected to hypothermia and then a lethal injection of sodium pentabarbitol prior to all experiments described in the following text. Retrograde dye perfusion combined with histological techniques demonstrated a compartmentalization of the proportionally large VNO from the main olfactory epithelium (MOE) in cryosections of L. bellii. SDS-PAGE analysis of the VNO of both species demonstrated the expression of three G protein subunits, namely, G(alphao), G(alphai2), and G(beta), and the absence of G(alphaolf), G(alpha11), and G(q), the latter of which are traditionally found in the MOE. Vomeronasal (VN) neurons were enzymatically isolated for whole cell voltage-clamp electrophysiology of single neurons. Both species demonstrated a tetrodotoxin (TTX)-sensitive, rapidly inactivating sodium current and a tetraethylammonium (TEA)-sensitive potassium current that had a transient and sustained component. VN neurons were classified into two types dependent on the ratio of sodium over sustained potassium current. VN neurons exhibited outward and inward chemosignal-evoked currents when stimulated with pheromone-containing secretions taken from the feces, skin, and precloacal pores. Fifty-nine percent of the neurons were responsive to at least one compound when presented with a battery of five different secretions. The breadth of responsiveness (H metric) demonstrated a heterogeneous population of tuning with a mean of 0.29.

Age and season affect chemical discrimination of Liolaemus bellii own space.

We explored chemical discrimination of own vs. novel space by different age classes (neonates, juveniles, and adults) of the lizard Liolaemus bellii, during pre- and post-hibernation seasons. We recorded the number of tongue flicks (TF) lizards produced during 10 min in their own or a novel enclosure. Age class and season affected chemical discrimination. Only adults and neonates discriminated their own space, albeit using different strategies: while adults made fewer TF in their own enclosure, neonates made more TF in their own enclosure. This difference was interpreted in terms of different requirements for discrimination of individuals during their lives. Increased chemical exploration by juveniles and adults at the onset of the post-hibernation season was associated with food-searching and reproductive behaviors.

Chemical composition of precloacal secretions of two Liolaemus fabiani populations: are they different?

The chemical composition of secretions of precloacal pores from two populations of the lizard Liolaemus fabiani (Puilar and Punta-Brava) were compared. This is an endemic species from the Atacama Salt Flat (26 degrees 46'S 68 degrees 14'W; 2400 m) in northern Chile, restricted to the internal lakes of the salt flat. Interpopulational differences in the chemical composition of the secretions were expected considering that populations have genetic differences and are subject to different thermal conditions. By using GC-MS, a total of 44 compounds were found belonging to three categories: n-alkanes, long-chain carboxylic acids, and steroids. Six compounds were found in all the individuals studied: tetradecanoic, pentadecanoic, hexadecanoic, hexadecenoic, octadecanoic, and octadecenoic acids. The secretions of both populations had similar types and proportions of the different compounds, except for cholesterol and hexanoic acid, which were more abundant in the Puilar population. These differences can be ascribed to the different environmental conditions to which populations are subjected. The adaptive meaning of these differences is discussed.