Unwrapping broken tails: Biological and environmental correlates of predation pressure in limbless reptiles.

Studying species interactions in nature often requires elaborated logistics and intense fieldwork. The difficulties in such task might hinder our ability to answer questions on how biotic interactions change with the environment. Fortunately, a workaround to this problem lies within scientific collections. For some animals, the inspection of preserved specimens can reveal the scars of past antagonistic encounters, such as predation attempts. A common defensive behaviour that leaves scars on animals is autotomy, the loss of a body appendage to escape predation. By knowing the collection site of preserved specimens, it is possible to assess the influence of organismal biology and the surrounding environment in the occurrence of autotomy. We gathered data on tail loss for 8189 preserved specimens of 33 snake and 11 amphisbaenian species to investigate biological and environmental correlates of autotomy in reptiles. We applied generalized linear mixed effect models to evaluate whether body size, sex, life-stage, habitat use, activity pattern, biome, tropicality, temperature and precipitation affect the probability of tail loss in limbless reptiles. We observed autotomy in 23.6% of examined specimens, with 18.7% of amphisbaenian and 33.4% of snake specimens showing tail loss. The probability of tail loss did not differ between snakes and amphisbaenians, but it was higher among large-sized specimens, particularly in adults and females. Chance of tail loss was higher for diurnal and arboreal species, and among specimens collected in warmer regions, but it was unaffected by biome, precipitation, and tropicality. Autotomy in limbless reptiles was affected by size-dependent factors that interplay with ontogeny and sexual dimorphism, although size-independent effects of life-stage and sex also shaped behavioural responses to predators. The increase in probability of tail loss with verticality and diurnality suggests a risk-balance mechanism between species habitat use and activity pattern. Although autotomy is more likely in warmer regions, it seems unrelated to seasonal differences in snakes and amphisbaenians activity. Our findings reveal several processes related to predator-prey interactions involving limbless reptiles, demonstrating the importance of scientific collections to unveil ecological mechanisms at different spatio-temporal scales.

Reproductive Biology, Sperm storage, and Sexual Maturity of Thamnodynastes strigatus (Serpentes: Dipsadidae).

Life history strategies determine and influence many aspects of species fitness. In this study, we describe the reproductive biology - reproductive cycle, sperm storage, and sexual maturity - of Thamnodynastes strigatus in South Brazil. We analyzed 49 individuals (25 males and 24 females) from herpetological collections. The reproductive cycle of males and females was described considering the morpho-anatomical and histological changes in the testes, ductus deferens, and kidney, as well in the ovary and oviduct. The age at the onset of sexual maturity was determined by skeletochronology of the caudal vertebra. The reproductive cycle is seasonal semi-synchronous and most individuals have a reproductive peak in spring and summer. The seasonal biennial reproductive cycle and viviparity are two phylogenetically conserved characters in Tachymenini snakes. Thamnodynastes Strigatus females store sperm in the utero-vaginal junction furrows during autumn. There were no differences between the ages of sexual maturity of males (4-11y) and females (4-12y). Females reach sexual maturity at larger body sizes, and this may confer an adaptive advantage due to a higher fecundity potential. Herein, we confirmed the previously described seasonal biennial reproductive cycle of T. strigatus through histological analysis.

First record of growth rings for 11 native subtropical anuran species of South America.

Skeletochronology is the most accurate method to estimate a population age structure. The methodology is based on the analysis of secondary bone tissue in order to count growth rings. This study aimed to provide initial data, allowing researchers to further work out in the age of individuals and populations, sampling evidence of the presence of growth rings in 11 native species (representing nine families) of a subtropical region of southern Brazil. Four bone samples of each specimen were used to perform the skeletochronological analysis: the penultimate phalanges of the 3rd and 4th fingers, the humerus, and the femur. The presence of growth rings was confirmed in the periosteal layer of the bones of all analyzed species. In comparison with phalanges, growth rings of humeri and femora are more irregular and less distinguishable. This is the first record of growth rings to the native species herein analised. The skeletochronology was proved to be an effective tool in determining the age of anuran amphibians from a subtropical region, since this environment presents well defined climatic seasonality.

Hematophagous Flies Attracted to Frog Calls in a Preserved Seasonal Forest of the Austral Neotropics, with a Description of a New Species of Corethrella (Diptera: Corethrellidae).

The signaler-eavesdropper interaction has been investigated for a wide range of organisms, and although many flies feed on calling frogs, this dynamic has been addressed only poorly in the austral Neotropics. We investigated this interaction in southern Brazil using pairs of suction traps (acoustic + silent) broadcasting frog calls or an artificial white noise in ponds and streams. From 139 sessions, flies of the genera Corethrella (Corethrellidae), Forcipomyia (Ceratopogonidae) and Uranotaenia (Culicidae) were collected, including five Corethrella species, the most abundant of which was previously unknown and is formally described here. Additionally, we present the southernmost records of Corethrella lopesi, C. alticola and C. atricornis. Numbers of Forcipomyia midges and Uranotaenia mosquitoes did not differ between silent traps and traps broadcasting frog calls, and did not differ between white noise traps and adjacent silent traps. However, the number of female Corethrella was significantly higher in traps broadcasting calls of the pond-breeding frog P. aff. gracilis compared to adjacent silent traps; calls of this frog attracted the five Corethrella species and also collected significantly more female Corethrella than the white noise. By evaluating different taxa of flies and broadcasting different sounds, we demonstrated that Corethrella midges were attracted only to the acoustic cue of P. aff. gracilis calls, while Forcipomyia and Uranotaenia were captured in traps by chance. Our results suggest that female Corethrella feed on males of the common pond-breeding frog P. aff. gracilis in southern Brazil, and highlight the utility of frog call traps in revealing the diversity of Corethrella in the austral Neotropics.

Reproductive phenology of the American Bullfrog in subtropical Brazil: photoperiod as a main determinant of seasonal activity.

The North American bullfrog Lithobates catesbeianus continues to invade ecosystems worldwide, potentially causing population declines and even extinctions. Within its native distribution, bullfrogs show prolonged reproductive seasons and high fertility. However, data on breeding biology of bullfrogs ex-situ in invaded localities mainly comes from anecdotal reports. Understanding how invasive species are adjusting their life histories to new colonized environments is important for conservation purposes. Here we describe temporal and spatial abundance, calling activity, spawning and tadpole distribution of bullfrogs in southern Brazil. Eighteen samplings occurred during one year. The abundance of individuals was positively related to longer photoperiods and higher temperatures. Reproductive activity was also positively associated with longer photoperiods. Calling sites, spawning and tadpoles were associated with microhabitats presenting hydrophytes, which may provide shelter and thermal stability to bullfrogs. The reproductive seasonal activity of bullfrogs can be highly variable across its growing geographical range, but in subtropical Brazil it is associated with photoperiod, a highly predictable abiotic determinant. In our study area, bullfrogs presented a breeding season twice as long as that observed in some native localities. We suggest that management strategies directed to bullfrog populations must consider the habitat structures and seasonal regimes determined by each invaded environment.

Molecular and sensory mechanisms to mitigate sunlight-induced DNA damage in treefrog tadpoles.

The increased incidence of solar ultraviolet B (UVB) radiation has been proposed as an environmental stressor, which may help to explain the enigmatic decline of amphibian populations worldwide. Despite growing knowledge regarding the UV-induced biological effects in several amphibian models, little is known about the efficacy of DNA repair pathways. In addition, little attention has been given to the interplay between these molecular mechanisms with other physiological strategies that avoid the damage induced by sunlight. Here, DNA lesions induced by environmental doses of solar UVB and UVA radiation were detected in genomic DNA samples of treefrog tadpoles (Hypsiboas pulchellus) and their DNA repair activity was evaluated. These data were complemented by monitoring the induction of apoptosis in blood cells and tadpole survival. Furthermore, the tadpoles' ability to perceive and escape from UV wavelengths was evaluated as an additional strategy of photoprotection. The results show that tadpoles are very sensitive to UVB light, which could be explained by the slow DNA repair rates for both cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6,4) pyrimidone photoproducts (6,4PPs). However, they were resistant to UVA, probably as a result of the activation of photolyases during UVA irradiation. Surprisingly, a sensory mechanism that triggers their escape from UVB and UVA light avoids the generation of DNA damage and helps to maintain the genomic integrity. This work demonstrates the genotoxic impact of both UVB and UVA radiation on tadpoles and emphasizes the importance of the interplay between molecular and sensory mechanisms to minimize the damage caused by sunlight.

Identification of influential events concerning the Antarctic ozone hole over southern Brazil and the biological effects induced by UVB and UVA radiation in an endemic treefrog species.

The increased incidence of solar ultraviolet radiation (UV) due to ozone depletion has been affecting both terrestrial and aquatic ecosystems and it may help to explain the enigmatic decline of amphibian populations in specific localities. In this work, influential events concerning the Antarctic ozone hole were identified in a dataset containing 35 years of ozone measurements over southern Brazil. The effects of environmental doses of UVB and UVA radiation were addressed on the morphology and development of Hypsiboas pulchellus tadpole (Anura: Hylidae), as well as on the induction of malformation after the conclusion of metamorphosis. These analyzes were complemented by the detection of micronucleus formation in blood cells. 72 ozone depletion events were identified from 1979 to 2013. Surprisingly, their yearly frequency increased three-fold during the last 17 years. The results clearly show that H. pulchellus tadpole are much more sensitive to UVB than UVA light, which reduces their survival and developmental rates. Additionally, the rates of micronucleus formation by UVB were considerably higher compared to UVA even after the activation of photolyases enzymes by a further photoreactivation treatment. Consequently, a higher occurrence of malformation was observed in UVB-irradiated individuals. These results demonstrate the severe genotoxic impact of UVB radiation on this treefrog species and its importance for further studies aimed to assess the impact of the increased levels of solar UVB radiation on declining species of the Hylidae family.

Seasonal, daily activity, and habitat use by three sympatric pit vipers (Serpentes, Viperidae) from southern Brazil.

Viperid snakes are widely distributed in the South America and the greater distribution range of the family is found at the Crotalinae subfamily. Despite the abundance of this snakes along their geographic distribution, some ecological aspects remain unknown, principally at subtropical areas. In the present study, we evaluated the activity (daily and seasonal) and the use of the habitat by Bothrops diporus, B. jararaca and B. jararacussu, in an Atlantic Forest area at southern Brazil. We observed higher incidence of viperid snakes during the months with higher temperatures, while no snakes were found during the months with lower temperatures. The data suggest the minimum temperature as environmental variable with the greatest influence on the seasonal activity of this species. Considering the daily activity, we observed a tendency of snakes to avoid the warmest hours. Bothrops jararacussu tend to avoid open areas, being registered only inside and at the edges of the forest. We compared our results with previous studies realized at tropical areas and we suggest the observed seasonal activity as an evolutive response, despite the influence of the different environmental variables, according to the occurence region.

The role of phytophysiognomies and seasonality on the structure of ground-dwelling anuran (Amphibia) in the Pampa biome, Southern Brazil.

Considering that habitat use by amphibians is related both with climate and environmental features, we tested the hypothesis that anuran assemblages found in different phytophysiognomies and in different seasons vary in structure. Additionally, we searched for species which can be indicators of habitat and seasons. The study was conducted in the Pampa biome, southern Brazil. Sampling was done through pitfall traps placed in three phytophysiognomies: grassland, ecotone grassland/forest; and forest. The seasonality factor was created by grouping months in warn and cold seasons. Sixteen species were found and the assemblages were influenced both by phytophysiognomies and climatic seasonality. In a paired comparison, the three phytophysiognomies differed in structure of assemblage from each other. Physalaemus henselii, P. riograndensis, Pseudopaludicola falcipes and Pseudis minuta were indicators of ecotone. Leptodactylus gracilis and Physalaemus biligonigerus were indicators of grassland. None species was indicator of forest. Most of the species were indicators of warm season: Elachistocleis bicolor, Leptodactylus fuscus, L. gracilis, L. latinasus, L. latrans, L. mystacinus, Physalaemus biligonigerus, P. cuvieri and Pseudis minuta. None species was indicator of cold season. We found that even for species of open areas, as Pampa, heterogeneous phytophysiognomies are important for maintaining abundance and constancy of populations of anuran.

Efficiency of snake sampling methods in the Brazilian semiarid region.

The choice of sampling methods is a crucial step in every field survey in herpetology. In countries where time and financial support are limited, the choice of the methods is critical. The methods used to sample snakes often lack objective criteria, and the traditional methods have apparently been more important when making the choice. Consequently researches using not-standardized methods are frequently found in the literature. We have compared four commonly used methods for sampling snake assemblages in a semiarid area in Brazil. We compared the efficacy of each method based on the cost-benefit regarding the number of individuals and species captured, time, and financial investment. We found that pitfall traps were the less effective method in all aspects that were evaluated and it was not complementary to the other methods in terms of abundance of species and assemblage structure. We conclude that methods can only be considered complementary if they are standardized to the objectives of the study. The use of pitfall traps in short-term surveys of the snake fauna in areas with shrubby vegetation and stony soil is not recommended.

Genotoxic risk assessment of solar UV radiation in tadpoles from Brazilian wetlands.

Solar ultraviolet (UV) radiation is an environmental genotoxic factor linked to amphibian decline. Here we assessed the genotoxic risk of UVB and UVA exposure for tadpoles from open ponds in southern Brazil, a mid-latitude region influenced by stratospheric ozone depletion. Daily UV doses were measured on the surface of a pond in Taim Ecological Station (TAIM; 32°49'24''S; 52°38'31''W) on a cloudless summer day to predict the worst-case scenario for UV-induced DNA damage. Pond descriptors were related to the use of microhabitats by Boana pulchella tadpoles in two ponds over the climate seasons of 2013 and 2014. Our results indicate that shaded microhabitats were more frequent than unshaded ones in autumn, winter, and spring but not in summer. Hence, the penetration of UV radiation into the water of unshaded microhabitats was evaluated through laboratory experiments with artificial UV sources and pond water samples. Physical and biological sensors were applied in the experiments to measure the incident UV radiation and its genotoxic action. By integrating field and laboratory data, we demonstrate that low doses of biologically effective UV radiation reached the tadpoles in autumn, winter, spring, and early summer due to a high proportion of shaded microhabitats and a high concentration of solids in unshaded microhabitats. However, the relative reduction of shaded microhabitats jointly with a declining water level in late summer may have exposed tadpoles to high UV doses. Our experiments also indicate that solar UVB radiation, but not UVA, is primarily responsible for the induction of DNA pyrimidine dimers in organisms living under the surface of aquatic ecosystems. The present work highlights the determinant role of wetland descriptors for minimizing the genotoxic potential of UV radiation and its consequences for amphibians.

Pampean lizard assemblage from subtropical Brazil: a temporal analysis.

The increasing human occupation of natural environments is one of the greatest threats to biodiversity. To mitigate the negative anthropogenic effects, it is necessary to understand the characteristics of natural populations and the natural history of species. A study was conducted with an assemblage of lizards from a disturbed area of the Pampa biome, from February 2001 to January 2004. The assemblage showed a unimodal seasonal pattern, with the recruitment period occurring during the warmer months. The captures were seasonal for two of the three monitored years, and concentrated within warmer months. The minimum temperature explained the number of catches for the assemblage as a whole. However, when the species were analyzed individually, the temperature only explained the seasonal occurrence of Teius oculatus. The abundance of species was significantly different in the third year of study for Cercosaura schreibersii and Ophiodes striatus. This latter species was no longer registered in the study area from May 2003 until the end of the study. Therefore, O. striatus may be more sensitive to environmental changes, considering the events of change in vegetation during the study. With frequent and increasing environmental disturbances, it is necessary to take conservation measures and encourage the increase of knowledge on Pampean lizards.

Pet distribution modelling: Untangling the invasive potential of Trachemys dorbigni (Emydidae) in the Americas.

Human activities have been changing the global biogeographic patterns by the introductions of invasive species. For reptiles, the invasion rate increase of non-native species is remarkably related to the pet trade, especially for freshwater turtles. Here we estimated the invasive potential of the South American turtle Trachemys dorbigni in the Americas using a combination of climatic and human activity variables. We built species distribution models based on data from the native and invasive ranges, using the ensemble model from five different algorithms (GAM, MAXENT, BRT, RF and GBM). We compared the two models' performance and predictions, one calibrated with only climatic variables (climate-driven), and the second also included a descriptive variable of human activity (climate plus human-driven). Suitable areas for T. dorbigni covered occurrence areas of its congeners and highly diversified ecoregions, such as the eastern USA, the islands of Central America, and the south eastern and eastern Brazilian coast. Our results indicate that human activities allow T. dorbigni to establish populations outside of its original climatic niche. Including human activity variables proved fundamental to refining the results to identify more susceptible areas to invasion and to allow the efficient targeting of prevention measures. Finally, we suggested a set of actions to prevent T. dorbigni becoming a highly impacting species in the areas identified as more prone to its invasion.

Correction: Effects of traffic noise on the calling behavior of two Neotropical hylid frogs.

[This corrects the article DOI: 10.1371/journal.pone.0183342.].

Sunlight-induced genotoxicity and damage in keratin structures decrease tadpole performance.

The increased incidence of solar ultraviolet (UV) radiation, an environmental genotoxic agent, due to ozone depletion or deforestation may help to explain the enigmatic decline of amphibian populations in specific localities. In this work, we evaluated the importance of DNA repair performed by photolyases to maintain the performance of treefrog tadpoles after acute and chronic treatments with environmental-simulated doses of solar UVB and UVA radiation. Immediately after UV treatments, tadpoles were exposed to a visible light source to activate photolyases or kept in dark containers. The biological effects of UV treatments were evaluated through morphological, histological, locomotor and survival analyzes of Boana pulchella tadpoles (Anura: Hylidae). The results indicate that tadpole body weight suffered influence after both UVB and UVA treatments, although the body length was bit affected. The locomotor performance of UVB-exposed tadpoles was significantly reduced. In addition, UVB radiation induced a severe impact on tadpole skin, as well as on keratinized structures of mouth (tooth rows and jaw), indicating that these should be important effects of solar UV radiation in the reduction of tadpole performance. Furthermore, photolyases activation was fundamental for the maintenance of tadpole performance after chronic UVB exposures, but it was relatively inefficient after acute exposures to UVB, but not to UVA radiation. Therefore, this work demonstrates how the UV-induced genotoxicity and structural alterations in the skin and oral apparatus affect tadpole performance and survival.

Effects of traffic noise on the calling behavior of two Neotropical hylid frogs.

Anthropogenic disturbance has been pointed to as one of the major causes of the world´s biodiversity crisis. Among them, noise pollution is a potential underestimated threat, projected to increase in the next decades accompanying urban expansion. Rising levels of noise pollution may result in negative impacts on species highly dependent on acoustic communication. Amphibians have long served as model organisms for investigating animal acoustic communication because their reproduction depends on transmitting and receiving acoustic signals. A few studies have investigated the effects of anthropogenic noise on anurans, but there is still limited knowledge on how it affects them. In this study, we test the effect of two intensities of traffic noise on calling males of two Neotropical treefrogs species. We expect to record more changes in call parameters, to avoid masking effect, at higher intensity noise treatments, and in the species with higher call/noise frequency overlap. We performed a set of field playback experiments exposing male frogs to road noise at two different intensities (65dB and 75dB). Focal species are Boana bischoffi (high call/noise frequency overlap) and B. leptolineata (low call/noise frequency overlap). Both species changed acoustic parameters during or after the exposure to traffic noise. Advertisement call rate of B. bischoffi decreased during road noise, and dominant frequency decreased over time. Call length of B. leptolineata increased or decreased, depending on the order of noise intensity. We also observed spatial displacement in both species, which moved away from the noise source. Our results provide evidence that traffic noise affects anuran calling behavior, and noise intensity is an important factor affecting how species respond.

Can Protected Areas with Agricultural Edges Avoid Invasions? The Case of Bullfrogs in the Southern Atlantic Rainforest in Brazil.

Bruno Madalozzo, Camila Both, and Sonia Cechin (2016) The American bullfrog, Lithobates catesbeianus, is one of the 100 most harmful invasive species of the planet. Climatic and topographic models predict that the Atlantic Forest regions of southern Brazil are favorable for the establishment of invasive bullfrog populations. The predicted increase of temperature and concentration of gases associated with the greenhouse effect will augment the vulnerability of protected areas of the Atlantic forest to bullfrog invasions in the coming years. In this study we investigated to what extent protected areas of the Atlantic Forest surrounded by anthropogenic landscapes are vulnerable to bullfrog invasions. We conducted surveys in 36 waterbodies located either in a protected area or in anthropogenically modified adjacent locations on a forest-edge-agriculture gradient. We collected data on abundance and breeding to identify the main descriptors (local and landscape variables) that explain the distribution of bullfrogs along this gradient. The variance partitioning analysis showed a strongest association of bullfrog abundance with local waterbody descriptors (area-depth-hydroperiod) and secondarily with a forest-edge-agriculture gradient, i.e., the landscape. The observed distribution pattern suggests that protected areas are likely to be invaded by bullfrogs. Therefore, management strategies should focus on both scales: landscape and waterbody. Supervising the construction of large (permanent or deep) waterbodies in edge habitats of the park and adjacent areas can be effective and agriculture and forest management could importantly complement the prevention of invasions.

Ecologia e história natural das serpentes de uma área de Caatinga no nordeste brasileiro / Ecology and natural history of snakes in a area of Caatinga in northeastern Brazil

We studied a snake assemblage from an area of Caatinga with shrub and tree vegetation to describe the natural history of the snake species. A total of 636 individuals among 22 species from four families were recorded. The distribution of species abundances is log-normal; and the composition presents typical species from the Caatinga, with Oxybelis aeneus (Wagler, 1824) and Philodryas nattereri Steindachner, 1870 being the most common. The natural history of each species is described based on information regarding activity patterns, diet, habitat use, reproduction, and defensive repertoire obtained during the study, as well as on information available in the literature. The study area is located in a priority region for conservation and our results emphasize that conservation policies should be implemented in the region.(au)

Pampean lizard assemblage from subtropical Brazil: a temporal analysis

The increasing human occupation of natural environments is one of the greatest threats to biodiversity. To mitigate the negative anthropogenic effects, it is necessary to understand the characteristics of natural populations and the natural history of species. A study was conducted with an assemblage of lizards from a disturbed area of the Pampa biome, from February 2001 to January 2004. The assemblage showed a unimodal seasonal pattern, with the recruitment period occurring during the warmer months. The captures were seasonal for two of the three monitored years, and concentrated within warmer months. The minimum temperature explained the number of catches for the assemblage as a whole. However, when the species were analyzed individually, the temperature only explained the seasonal occurrence of Teius oculatus. The abundance of species was significantly different in the third year of study for Cercosaura schreibersii and Ophiodes striatus. This latter species was no longer registered in the study area from May 2003 until the end of the study. Therefore, O. striatus may be more sensitive to environmental changes, considering the events of change in vegetation during the study. With frequent and increasing environmental disturbances, it is necessary to take conservation measures and encourage the increase of knowledge on Pampean lizards.

O crescimento da ocupação humana sobre ambientes naturais é uma das maiores ameaças à biodiversidade. Para amenizar os efeitos negativos antropogênicos, é necessário entender as características das populações naturais, e a história natural das espécies. Um estudo foi conduzido com uma assembeia de lagartos de uma área perturbada do Pampa, de fevereiro de 2002 a janeiro de 2004. A assembleia apresentou padrão sazonal unimodal, com recrutamento ocorrendo durante os meses mais quentes. As capturas foram sazonais durante dois dos três anos monitorados, e concentradas nos meses mais quentes. A temperatura mínima explica o número de capturas para a assembleia como um todo. Entretanto, quando as espécies foram analisadas individualmente, a variável explica apenas a ocorrência sazonal de Teius oculatus. A abundância das espécies foi significativamente distinta no terceiro ano de estudo para Cercosaura schreibersii e Ophiodes striatus. Essa última espécie não teve mais registros na área de maio de 2003 até o final do estudo. Assim, O. striatus pode ser mais sensível às udanças abientais, considerando os eventos de udança na vegetação ocorridas durante o estudo. Com as frequentes e crescentes perturbações ambientais que o bioma vem sofrendo, são necessárias ações de conservação, e dar suporte para o aumento do conhecimento dos lagartos pampeanos.