Efeito de armadilha das passagens em grelha sobre os anfíbios

As passagens em grelha são infra-estruturas largamente utilizadas para impedir a deslocação do gado, mas que permitem a circulação de viaturas. Porém, também resultam numa armadilha de fosso onde pequenos animais terrestres podem cair inadvertidamente. O principal objectivo deste estudo foi verificar o efeito que as passagens em grelha têm sobre as populações de anfíbios, aquando das suas deslocações sazonais. Estudaram-se 18 passagens em grelha localizadas em duas zonas de amostragem - Elvas e Alandroal - no Alto Alentejo (Portugal). Esta investigação revelou que a principal espécie encarcerada nestas infra-estruturas é o sapo-de-unha-negra (Pelobates cultripes). Em termos gerais, os anfíbios mais terrestres são significativamente (teste do x2) mais afectados do que os anfíbios marcadamente aquáticos. O teste de Mantel mostrou que não existe correlação entre o tipo de passagem em grelha e os anfíbios encontrados nessas infra-estruturas. Para mitigar o aprisionamento de anfíbios nas passagens em grelha, são propostas cinco medidas. ABSTRACT; Cattle grids are infrastructures widely used to avoid the movement of cattle along a road, though allowing the circulation of vehicles. However, they also result in a pit trap where small terrestrial animals may inadvertently fall. The main objective of this study was to check the effect that cattle grids have on amphibian populations, during their seasonal migrations. 18 cattle grids, located in two sampling zones - Elvas and Alandroal- in Alto Alentejo (Portugal), were studied. This investigation showed that the main species imprisoned in these devices is the iberian spadefoot toad (Pelobates cultripes). Generally, the more terrestrial amphibians are significantly (x2 test) more affected than markedly aquatic amphibians. Mantel test showed that there is no correlation between different kinds of cattle grids and the amphibians found in these infrastructures. To mitigate the imprisonment of amphibians in cattle grids, five measures are proposed.

Estudo da mortalidade de anfíbios em duas estradas rurais próximas: efeito do tráfego rodoviário

As rodovias podem contribuir para a diminuição de populações de anfíbios de várias formas, sendo a mais visível a mortalidade por atropelamento. Neste trabalho procurou-se observar a forma como o tráfego pode afectar a mortalidade de anfíbios. A recolha de dados foi efectuada em duas estradas rurais próximas onde foram registadas as espécies observadas e se estas estavam vivas ou mortas, entre outros. Os resultados comprovaram não haver diferenças significativas em relação ao número de indivíduos ou à proporção das espécies que atravessam as vias e confirmaram que uma das estradas possui uma percentagem de mortalidade muito superior à outra. Os testes estatísticos também revelaram existir uma diferença significativa entre o tráfego nas duas estradas. Desta forma, na estrada com mais trânsito foi observado um número muito superior de mortos. Foi, portanto, possível relacionar os resultados e concluir que os anfíbios são negativamente afectados pela intensidade do trânsito rodoviário. ABSTRACT; The roads can contribute to the amphibian populations decline in many ways, being the most visible the roadkills. The purpose of this study was to observe how the traffic can affect the amphibian mortality. The data collection was conducted in two rural roads, near to each other, where the species observed were recorded, and if they were alive or dead, among other information. The results showed no significant differences in the number of individual or in the proportion of species that cross the roads and confirmed that one of the road has a much higher mortality percentage than the other. Statistical tests also revealed a significant difference between the traffic of the two roads. Thus, on the road with more traffic was observed a much higher number of deaths. It was therefore possible to relate the results and conclude that the amphibians are negatively affected by the traffic intensity.

Community- and population-level effects of chytridiomycosis in a Neotropical amphibian community

As the number of fungal pathogen outbreaks become more frequent worldwide across taxa, so have the number of species extirpations and communities persisting with the pathogen. This phenomenon raises questions, such as: “what leads to host extinction during an outbreak?” and “how are hosts persisting once the pathogen establishes?.” But the data on host populations and communities across life stages before and after pathogen arrival rarely exist to answer these questions. Over the past three to four decades, the amphibian-killing fungus Batrachochytrim dendrobatidis (Bd) spread in a wave-like manner across Central America, leading to rapid species extirpations and population declines. I collected data on tadpole and adult amphibians in El Copé, Panama before, during, and after the Bd outbreak to answer these questions. I used Bayesian statistical approaches to account for imperfect host and pathogen detection of marked and unmarked individuals. In the tadpole community, within 11 months of Bds arrival, density and occupancy rapidly declined. Species losses were phylogenetically correlated, with glass frogs disappearing first, and tree frogs and poison-dart frogs remaining. I found that tadpole communities resembled one another more strongly after the outbreak than they did before Bd invasion. I found no tadpoles within 22 months of the outbreak and limited signs of recovery within 10 years. In contrast, at the same site, for a population of male glass frogs, Espadarana prosopleon, I found that 10 years post-outbreak, the population was consistently half its historic abundance, and that the lack of recruits to the population explained why the population had not rebounded, rather than high pathogen-induced mortality. And finally, examining the entire amphibian community, I found high pathogen prevalence, low infection intensities, and high survival rates of uninfected and infected hosts. Bd transmission risk, i.e., the probability a susceptible host becomes infected, did not relate to host density, pathogen prevalence, or infection intensity– Bd transmission risk was uniform across the study area. My results are especially relevant to conservation biologists aiming to predict the future impacts of Bd outbreaks, those trying to manage persisting populations, and those interested in reintroducing species back into wild amphibian communities.

Síntese química e avaliação das propriedades antibacterianas e antiparasitárias de análogos de peptídeos antimicrobianos de anuros

Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Molecular, 2015.

The role of forest certification for the conservation of biodiversity and sustainability of cork oak woodlands

Doutoramento em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia - UL

Estabelecimento de prioridades para a conservação dos vertebrados terrestres do Parque Natural da Ria Formosa e selecção de áreas para a sua protecção

Dissertação de mest. em Gestão e Conservação da Natureza, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2004

Landscape context influences chytrid fungus distribution in an endangered European amphibian

Wildlife disease is an emerging threat to biodiversity. The amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd), which causes the disease chytridiomycosis, has been documented in over 500 amphibian species globally. Understanding conditions under which amphibians are vulnerable to Bd is important for evaluating species risk and developing surveillance strategies. Here, we investigate the spatial distribution of Bd infection in the ephemeral pond-breeding yellow-bellied toad Bombina variegata, a species of high conservation concern in the European Union. We sampled 550 toads from 60 ponds in a traditional agricultural landscape in Southern Transylvania, Romania. Overall, Bd prevalence was low in B.variegata, but infected toads were widely dispersed through the landscape and were found in a quarter of all sampled ephemeral ponds. At the pond level, increased Bd occurrence was associated with short distances to perennial water sources and high forest cover. These findings suggest that perennial water sources may act as source habitat for Bd, with amphibian movements resulting in Bd spillover into ephemeral ponds. Increased Bd occurrence in ponds surrounded by high levels of forest cover is likely related to cooler and wetter conditions that are more favourable for Bd. Throughout the study landscape, patchy environmental suitability for Bd appears to restrict the pathogen to a subset of B.variegata habitat. Ephemeral ponds in open landscapes, without nearby perennial habitat, likely provide an environmental refuge from Bd, where the risk of infection is low. From a conservation perspective, these findings highlight the importance of maintaining ephemeral ponds in open landscapes, but these are currently threatened by land-use change.

Widespread convergence in toxin resistance by predictable molecular evolution

The question about whether evolution is unpredictable and stochastic or intermittently constrained along predictable pathways is the subject of a fundamental debate in biology, in which understanding convergent evolution plays a central role. At the molecular level, documented examples of convergence are rare and limited to occurring within specific taxonomic groups. Here we provide evidence of constrained convergent molecular evolution across the metazoan tree of life. We show that resistance to toxic cardiac glycosides produced by plants and bufonid toads is mediated by similar molecular changes to the sodium-potassium-pump (Na(+)/K(+)-ATPase) in insects, amphibians, reptiles, and mammals. In toad-feeding reptiles, resistance is conferred by two point mutations that have evolved convergently on four occasions, whereas evidence of a molecular reversal back to the susceptible state in varanid lizards migrating to toad-free areas suggests that toxin resistance is maladaptive in the absence of selection. Importantly, resistance in all taxa is mediated by replacements of 2 of the 12 amino acids comprising the Na(+)/K(+)-ATPase H1-H2 extracellular domain that constitutes a core part of the cardiac glycoside binding site. We provide mechanistic insight into the basis of resistance by showing that these alterations perturb the interaction between the cardiac glycoside bufalin and the Na(+)/K(+)-ATPase. Thus, similar selection pressures have resulted in convergent evolution of the same molecular solution across the breadth of the animal kingdom, demonstrating how a scarcity of possible solutions to a selective challenge can lead to highly predictable evolutionary responses.

First molecular determination of herpesvirus from two mysticete species stranded in the Mediterranean Sea

BACKGROUND Herpesvirus can infect a wide range of animal species: mammals, birds, reptiles, fish, amphibians and bivalves. In marine mammals, several alpha- and gammaherpesvirus have been identified in some cetaceans and pinnipeds species. To date, however, this virus has not been detected in any member of the Balaenoptera genus. CASE PRESENTATION Herpesvirus was determined by molecular methods in tissue samples from a male fin whale juvenile (Balaenoptera physalus) and a female common minke whale calf (Balaenoptera acutorostrata) stranded on the Mediterranean coast of the Region of Valencia (Spain). Samples of skin and penile mucosa from the fin whale and samples of skin, muscle and central nervous system tissue from the common minke whale tested positive for herpesvirus based on sequences of the DNA polymerase gene. Sequences from fin whale were identical and belonged to the Alphaherpesvirinae subfamily. Only members of the Gammaherpesvirinae subfamily were amplified from the common minke whale, and sequences from the muscle and central nervous system were identical. Sequences in GenBank most closely related to these novel sequences were viruses isolated from other cetacean species, consistent with previous observations that herpesviruses show similar phylogenetic branching as their hosts. CONCLUSIONS To our knowledge, this is the first molecular determination of herpesvirus in the Balaenoptera genus. It shows that herpesvirus should be included in virological evaluation of these animals.

Toxicity of polychlorinated biphenyls and N-phenyl-1-naphthylamine in the juvenile turtle, Chelydra serpentina

Polychlorinated biphenyls (PCBs) and substituted phenylamine antioxidants (SPAs) are two chemical groups that have been used in multiple Canadian industrial processes. Despite the production ban of PCBs in North America in 1977, they are still ubiquitous in the environment and in wildlife tissues. Previous studies of fish, amphibians, birds, and mammals have shown that PCBs are toxic and act as endocrine disruptors. In contrast, SPAs, specifically N-phenyl-1-naphthylamine (PANA), have received very little attention despite their current use in Canada and their expected environmental releases. The effects of PCB and PANA exposures in reptiles remain unknown thus, juvenile Chelydra serpentina were used in this thesis as a model vertebrate to fill in missing toxicity research gaps due to their importance as an environmental indicator. First, food pellets were spiked at an environmentally relevant concentration of the PCB mixture Aroclor 1254 (A1254) to model hepatic bioaccumulation (0.45 μg/g A1254 for 31 days) and depuration (clean food for 50 days) of PCBs in turtles. No significant differences in PCB concentrations were observed between the control and treated animals, suggesting that juvenile turtles exposed to environmentally relevant concentrations of PCBs can likely detoxify low concentrations of PCBs. Additionally, two dose-response experiments were performed using A1254 or PANA spiked food (0-12.7 μg/g and 0-3,446 μg/g, respectively) to determine hepatic toxicity and bioaccumulation in juvenile C. serpentina. An increase in hepatic cyp1a was observed when exposed to the highest dose of both chemicals: 1) for A1254, induction correlated to the significant increase in hepatic PCB congeners that are known to be metabolized by CYP1A; and 2) for PANA, induction suggested that CYP1A has a potential role in its detoxification. PCBs are known endocrine disruptors, but no significant changes were observed for both thyroid receptors (alpha and beta) or by estrogen and androgen receptors. This lack of response, also noted in the PANA exposure, suggests that C. serpentina is less sensitive to endocrine disruption than other vertebrates. Furthermore, the expression of genes involved in cellular stress was not altered in PCB and PANA exposed animals, supporting the resilience of turtles to oxidative stress. This is the first study to demonstrate the toxicity of PCBs and PANA in C. serpentina, demonstrating the turtle’s high tolerance to contamination.

Edge enhancement improves disruptive camouflage by emphasising false edges and creating pictorial relief

Disruptive colouration is a visual camouflage composed of false edges and boundaries. Many disruptively camouflaged animals feature enhanced edges; light patches are surrounded by a lighter outline and/or a dark patches are surrounded by a darker outline. This camouflage is particularly common in amphibians, reptiles and lepidopterans. We explored the role that this pattern has in creating effective camouflage. In a visual search task utilising an ultra-large display area mimicking search tasks that might be found in nature, edge enhanced disruptive camouflage increases crypsis, even on substrates that do not provide an obvious visual match. Specifically, edge enhanced camouflage is effective on backgrounds both with and without shadows; i.e. this is not solely due to background matching of the dark edge enhancement element with the shadows. Furthermore, when the dark component of the edge enhancement is omitted the camouflage still provided better crypsis than control patterns without edge enhancement. This kind of edge enhancement improved camouflage on all background types. Lastly, we show that edge enhancement can create a perception of multiple surfaces. We conclude that edge enhancement increases the effectiveness of disruptive camouflage through mechanisms that may include the improved disruption of the object outline by implying pictorial relief.

Reservoir-host amplification of disease impact in an endangered amphibian

Emerging wildlife pathogens are an increasing threat to biodiversity. One of the most serious wildlife diseases is chytridiomycosis, caused by the fungal pathogen, Batrachochytrium dendrobatidis (Bd), which has been documented in over 500 amphibian species. Amphibians vary greatly in their susceptibility to Bd, with some species tolerating infection, while others experience rapid mortality. Reservoir hosts - species that carry infection while maintaining high abundance, but are rarely killed by disease - can increase extinction risk in highly susceptible, sympatric species. However, whether reservoir hosts amplify Bd in declining amphibian species has not been examined. We combine a laboratory study with field surveys, disease sampling, and statistical modeling to investigate the role of reservoir hosts in chytridiomycosis dynamics and species decline in an amphibian community in south-eastern Australia. We show that the non-declining common eastern froglet (Crinia signifera) is a reservoir host for Bd, with laboratory animals carrying intense infection burdens over 12 weeks and the majority of wild sampled individuals carrying intense infections. We find that the presence of C. signifera is strongly associated with Bd prevalence in the sympatric, IUCN red-listed northern corroboree frog (Pseudophryne pengilleyi). Consistent with disease amplification by a reservoir host, we find that P. pengilleyi has declined from areas with high C. signifera abundance. Our results suggest that when reservoir hosts are present, population declines can continue long after the initial emergence of Bd, highlighting an urgent need to assess extinction risk in remnant populations of other declined amphibian species. Reintroductions and in situ management strategies must focus on identifying reservoir hosts and minimizing exposure of threatened species.

Limited influence of stream networks on the terrestrial movements of three wetland-dependent frog species

Quantifying functional connectivity is essential for understanding factors that limit or promote animal dispersal in fragmented landscapes. Topography is a major factor influencing the movement behavior of many animal species, and therefore the extent of functional connectivity between habitat patches. For pond-breeding frogs, areas of low topographic relief (such as streams or drainage lines) offer damp microhabitats that can facilitate movement through otherwise dry landscapes. However, the extent of topographic bias of frog movements has rarely been quantified. We used a replicated study to compare captures in high- and low-relief transects, for three species from a pond-breeding frog community in southeastern Australia. We captured frogs significantly more often on low-relief transects. However, capture rates decreased with increasing distance from water at similar rates on both high-relief and low-relief transects, and we observed few differences between adult and juvenile movements. Our results suggest that although low-relief drainage lines are important for the pond-breeding frogs in question, ecologists and landscape managers should not discount the role of high-relief locations. Because low-relief drainage lines represent a low proportion of the pond margin, >90% of movements are likely to occur across high-relief locations. Therefore, for the species that we studied, buffer zones designed to conserve only hydrological networks would provide insufficient protection of frequently used pond margins, while drainage lines are unlikely to act as vital networks facilitating connectivity between breeding ponds. Our study suggests that movement across slopes may be most important for facilitating functional connectivity.

Distinguishing the distributions of two cryptic frogs (Anura: Discoglossidae) using molecular data and environmental modeling

Currently, the identification of two cryptic Iberian amphibians, Discoglossus galganoi Capula, Nascetti, Lanza, Bullini and Crespo, 1985 and Discoglossus jeanneae Busack, 1986, relies on molecular characterization. To provide a means to discern the distributions of these species, we used 385-base-pair sequences of the cytochrome b gene to identify 54 Spanish populations of Discoglossus. These data and a series of environmental variables were used to build up a logistic regression model capable of probabilistically designating a specimen of Discoglossus found in any Universal Transverse Mercator (UTM) grid cell of 10 km × 10 km to one of the two species. Western longitudes, wide river basins, and semipermeable (mainly siliceous) and sandstone substrates favored the presence of D. galganoi, while eastern longitudes, mountainous areas, severe floodings, and impermeable (mainly clay) or basic (limestone and gypsum) substrates favored D. jeanneae. Fifteen percent of the UTM cells were predicted to be shared by both species, whereas 51% were clearly in favor of D. galganoi and 34% were in favor of D. jeanneae, considering odds of 4:1. These results suggest that these two species have parapatric distributions and allow for preliminary identification of potential secondary contact areas. The method applied here can be generalized and used for other geographic problems posed by cryptic species.

Assessing the conservation status of Mediterranean temporary ponds using biodiversity: a new tool for practitioners

The assessment of the habitat condition is the first step of conservation actions and several tools are available to assess wetlands. However, only a few tools are adapted to the priority habitat Mediterranean temporary ponds. Thus, our objectives were (i) to identify biological indicators associated with the different conservation status of Mediterranean tem- porary ponds and (ii) to create an efficient evaluation tool for non-experts using indicators of conservation status. A total of 87 ponds were sampled in southwest Portugal to assess the presence of plants, large branchiopods, amphibians, threatened voles and bats. Ponds with favourable conservation status showed higher species richness of plants, large branchiopods and amphibians. We identified eighteen indicators for favourable ponds: 15 plants, one large branchiopod and two amphibian taxa. We propose a new tool to assess the conservation status of Mediterranean tem- porary ponds based on the presence of these indicators. This tool is an alternative to other common, but time- consuming, methods and can be readily used by trained practitioners. The replication and adaptation of this tool to other regions and habitats enables the collection of comparable data and the geographical scaling-up of the assessments.