Investigation of amphibian mortality events in wildlife reveals an on-going ranavirus epidemic in the North of the Netherlands

In the four years following the first detection of ranavirus (genus Ranavirus, family Iridoviridae) infection in Dutch wildlife in 2010, amphibian mortality events were investigated nationwide to detect, characterize and map ranaviruses in amphibians over time, and to establish the affected host species and the clinico-pathological presentation of the disease in these hosts. The ultimate goal was to obtain more insight into ranavirus disease emergence and ecological risk. In total 155 dead amphibians from 52 sites were submitted between 2011 and 2014, and examined using histopathology, immunohistochemistry, virus isolation and molecular genetic characterization. Ranavirus-associated amphibian mortality events occurred at 18 sites (35%), initially only in proximity of the 2010 index site. Specimens belonging to approximately half of the native amphibian species were infected, including the threatened Pelobates fuscus (spadefoot toad). Clustered massive outbreaks involving dead adult specimens and ranavirus genomic identity indicated that one common midwife toad virus (CMTV)-like ranavirus strain is emerging in provinces in the north of the Netherlands. Modelling based on the spatiotemporal pattern of spread showed a high probability that this emerging virus will continue to be detected at new sites (the discrete reproductive power of this outbreak is 0.35). Phylogenetically distinct CMTV-like ranaviruses were found in the south of the Netherlands more recently. In addition to showing that CMTV-like ranaviruses threaten wild amphibian populations not only in Spain but also in the Netherlands, the current spread and risk of establishment reiterate that understanding the underlying causes of CMTV-like ranavirus emergence requires international attention

Dermocystid infection and associated skin lesions in free-living palmate newts (Lissotriton helveticus) from Southern France

Since the early 1900s, mesomycetozoan parasites have been reported in both European anuran and caudate species. These reports have primarily been descriptive, which has made assessing the impact of these parasites on host populations difficult. Anecdotal reports of Dermocystidium-like parasites are becoming widespread across Europe, possibly indicating that these mesomycetozoan parasites are increasing in distribution and/or abundance. This highlights the need for further investigations into the occurrence, pathogenesis and effects on host health of these parasitic infections for free-living amphibian populations, particularly those which are already stressed or threatened by other factors. Here we report the results of pathological, microbiological and molecular investigations used to characterize unidentified skin lesions in palmate newts (Lissotriton helveticus) from Larzac, France. We confirm that the lesions are the result of infection with a novel dermocystidium-like parasite, which is related to Amphybiocystidium ranae. We also show that the same parasite is distributed across several newt breeding sites. The lesions that result from infection with this parasite range from single or few vesicular or nodular cutaneous lesions to multiple coalescing skin ulcers with extensive hemorrhages. The latter have not been previously described in amphibians due to mesomycetozoan parasitic infection. Dermocystid DNA was detected only in newts that showed lesions, providing comparative evidence of the parasite's pathogenicity. We discuss the potential significance of the presence of this pathogen in the context of the population health of palmate newts

Anfibios y Reptiles del Museo de Zoología de la Universidad de Navarra

Se presenta el Catálogo actualizado de la Colección de Anfibios y Reptiles del Museo de Zoología de la Universidad de Navarra. Una pequeña parte de la colección se halla expuesta al público y forma parte del Museo de Ciencias Naturales de la Universidad de Navarra. A fecha de edición, la colección completa incluye más de 2500 ejemplares, de los que dos tercios son anfibios. Entre otras, 50 especies ibéricas están representadas en la colección

Heritability of Batrachochytrium dendrobatidis burden and its genetic correlation with development time in a population of Common toad (Bufo spinosus)

Despite the important threat that emerging pathogens pose for the conservation of biodiversity as well as human health, very little is known about the adaptive potential of host species to withstand infections. We studied the quantitative genetic architecture responsible for the burden of the fungal pathogen Batrachochytrium dendrobatidis in a population of common toads in conjunction with other life-history traits (i.e., body size and development rate) that may be affected by common selective pressures. We found a significant heritable component that is associated with fungal burden, which may allow for local adaptation to this pathogen to proceed. In addition, the high genetic correlation found between fungal burden and development time suggests that both traits have to be taken into account in order to assess the adaptive response of host populations to this emerging pathogen.Our research was supported by the following grants: Spanish Ministry of Education reference CGL2011-23443, Ministry of Competitivity and Economy reference BES-2012-055220, and Spanish Organization of National Parks reference MARM 428/211.Peer reviewe

Extreme reduction in body size and reproductive output associated with sandy substrates in two anuran species

Geographic variation in body size and reproductive traits has been reported in a wide range of organisms, including amphibians. Most studies have focused on latitudinal and/or altitudinal variation where differences in temperature and duration of the growing season are the main causes for population divergence. We describe a steep variation in body size and reproductive traits in two anuran species in southwestern Spain, associated with changes in the geological substrate. Pelobates cultripes and Bufo calamita (= Epidalea calamita) drastically reduced their size (a 71.6% and 76.1% reduction in body mass for P. cultripes and B. calamita, respectively) in just about 60 km. This extreme size reduction was more pronounced at the boundary between two different geological substrates (hercinic and sandy soil). Mean clutch mass, egg size, and clutch size were all smaller in B. calamita populations in the sandy environment. Likewise, clutch mass and egg size were both smaller in sandy P. cultripes populations. We observed a negative correlation between size-adjusted fecundity and egg size for both species, suggesting the existence of a reproductive trade-off that could explain the differences in reproductive allocation between populations and species. In P. cultripes, small-bodied populations had relatively higher fecundities and smaller eggs than large-bodied ones, whereas in B. calamita populations from the sandy area we found both populations with high fecundity and small eggs, and populations with low fecundity and large eggs. Common environmental effects associated with the sandy substrate produce a similar reduction in size in both toad species. © 2008 Koninklijke Brill NV, Leiden.Peer Reviewe

Influencia local y paisajística sobre la riqueza de especies de anfibios que se reproducen en charcas temporales del litoral sud‐atlántico español. Determinación de impactos

The increase in urban land and the continuous increment of road network experienced by littoral zones contribute to the loss, degradation and isolation of both terrestrial and aquatic habitats in which amphibians spend the different stages of their life cycle. The aim of this study is to explain the relationship between the characteristics of 17 seasonal ponds located in the Atlantic littoral of Cádiz province (SW Spain), the land uses in their surroundings, the road network, and the diversity of amphibian species. Our results show that amphibian richness is higher in larger ponds with longer hydroperiods, surrounded by forest and scrubland, and away from the nearest road. Roads, and in particular secondary roads, have a great and negative impact on amphibian richness because the shorter the distance between the pond and the road was, the fewer breeding species were found. From the data obtained in this study, we have classified the ponds according to their status of conservation. This classification demonstrates that 94% of the studied ponds require immediate measures to be taken in order to reduce the negative impact of habitat fragmentation caused by roads. Finally, in order to maintain healthy amphibian populations at the long term, forests and scrublands surrounding ponds must be protected, and connectivity among ponds, as well as with the terrestrial habitats in the vicinity, must be ensured. Preventing amphibians from road kills during their migration movements can be achieved by means of constructing underpasses and tunnels in hot spots.La proliferación de zonas urbanizadas y la expansión de la red de carreteras en zonas costeras contribuyen a la pérdida, degradación y aislamiento de los hábitats tanto terrestres como acuáticos utilizados por los anfibios en las diferentes fases de su ciclo vital. El objetivo de este estudio es explicar la relación entre las características de 17 charcas temporales ubicadas en el litoral atlántico de la provincia de Cádiz (SO de España), los usos del suelo en su entorno, la red de carreteras y la diversidad de especies de anfibios. Nuestros resultados muestran que la riqueza de anfibios es mayor en charcas más grandes y con hidroperiodos más prolongados, rodeadas de zonas forestales y de matorral, y alejadas de las carreteras. Las carreteras, en particular las secundarias, tienen un fuerte impacto negativo sobre la riqueza de anfibios, como muestra el hecho de que cuanto menor es la distancia entre la charca y la carretera, menor es el número de especies de anfibios presentes. A partir de los datos obtenidos, clasificamos las charcas de acuerdo a su estado de conservación. Esta clasificación demuestra que el 94% de las charcas estudiadas requiere la toma inmediata de medidas con el fin de reducir el impacto negativo de la fragmentación del hábitat causada por las carreteras. Por último, para mantener poblaciones de anfibios saludables a largo plazo, es preciso proteger los bosques y zonas de matorral del entorno de las charcas, así como la conectividad entre charcas y con el medio terrestre cercano. La construcción de túneles y pasos subterráneos en puntos negros de atropello puede servir para prevenir la mortalidad de anfibios en las carreterasPeer reviewe

Competition may explain the fine-scale spatial patterns and genetic structure of two co-occurring plant congeners

1. The spatial distribution of individual plants within a population and the population’s genetic structure are determined by several factors, like dispersal, reproduction mode or biotic interactions. The role of interspecific interactions in shaping the spatial genetic structure of plant populations remains largely unknown. 2. Species with a common evolutionary history are known to interact more closely with each other than unrelated species due to the greater number of traits they share. We hypothesize that plant interactions may shape the fine genetic structure of closely related congeners. 3. We used spatial statistics (georeferenced design) and molecular techniques (ISSR markers) to understand how two closely related congeners, Thymus vulgaris (widespread species) and T. loscosii (narrow endemic) interact at the local scale. Specific cover, number of individuals of both study species and several community attributes were measured in a 10 × 10 m plot. 4. Both species showed similar levels of genetic variation, but differed in their spatial genetic structure. Thymus vulgaris showed spatial aggregation but no spatial genetic structure, while T. loscosii showed spatial genetic structure (positive genetic autocorrelation) at short distances. The spatial pattern of T. vulgaris’ cover showed significant dissociation with that of T. loscosii. The same was true between the spatial patterns of the cover of T. vulgaris and the abundance of T. loscosii and between the abundance of each species. Most importantly, we found a correlation between the genetic structure of T. loscosii and the abundance of T. vulgaris: T. loscosii plants were genetically more similar when they were surrounded by a similar number of T. vulgaris plants. 5. Synthesis. Our results reveal spatially complex genetic structures of both congeners at small spatial scales. The negative association among the spatial patterns of the two species and the genetic structure found for T. loscosii in relation to the abundance of T. vulgaris indicate that competition between the two species may account for the presence of adapted ecotypes of T. loscosii to the abundance of a competing congeneric species. This suggests that the presence and abundance of close congeners can influence the genetic spatial structure of plant species at fine scales

Mediterranean temporary ponds as amphibian breeding habitats: The importance of preserving pond networks

We describe the spatial and temporal variation in the main characteristics of amphibian breeding habitats in Doñana National Park (south of Spain), during two hydrologic cycles with different rainfall amounts and timing (2002-2003 and 2005-2006). We also evaluate amphibian habitat requirements following a model selection approach based on Akaike's Information Criterion. Our results evidenced large spatial variability in all pond characteristics and inter-annual differences in pond hydroperiod, depth, and most water-chemistry characteristics. We observed a remarkable independence of pond characteristics at different sampling dates, suggesting that a pond description based on a single survey may not be representative. Eight of nine amphibian species attempted breeding in both years in spite of the marked inter-annual variability. Habitat models were species-specific and year-specific, as we found inter-annual differences in the pond characteristics relevant for species richness or for the relative abundance of particular species. All these results suggest that this large and diverse network of ponds provides different habitat opportunities each year, favouring the long-term persistence of the whole amphibian community. © Springer Science+Business Media B.V. 2009.Peer Reviewe

A new Late Pleistocene non-anthropogenic vertebrate assemblage from the northern Iberian Peninsula: Artazu VII (Arrasate, Basque Country)

Late Pleistocene palaeontological sites without human intervention are limited in the Cantabrian region, and even more so those with a good state of preservation and rich biodiversity. A new vertebrate fossiliferous locality discovered at Kobate Quarry (Arrasate, northern Iberian Peninsula) is presented in this article. This site, in which remains of 40 different vertebrate taxa were accumulated, acted as a natural trap. The preliminary vertebrate faunal list includes five amphibian taxa, four reptiles, seven species of birds and 24 mammalian taxa. While small mammals are represented by 13 small mammal taxa (seven in the Order Rodentia, five in the Order Eulipotyphla, and one in the Order Chiroptera), the large mammal fauna comprises eleven species, including ungulates and carnivores. The palaeoecology inferred from this faunal assemblage suggests the existence of large forested areas with some grassland and a watercourse nearby, within a notably warm and humid climate. These palaeoenvironmental conditions, combined with AMS and AAR results carried out in macrofaunal bone samples, suggest that the deposit from Artazu VII would be located in the first half of the Late Pleistocene, in the Marine Isotope Stage (MIS) 5c. Résumé Les gisements paléontologiques de la fin du Pléistocène sont limités dans la région Cantabrique, et encore plus limités sont les sites ayant fourni des restes osseux en bon état de conservation et montrant une grande biodiversité. Nous présentons ici un nouveau site trouvé dans la carrière de Kobate (Arrasate, Nord de la péninsule Ibérique). Ce site a certainement agi comme un piège naturel et contient une accumulation de restes appartenant à 40 espèces différentes de vertébrés. La liste préliminaire de la faune de vertébrés comprend cinq taxons d’amphibiens, quatre de reptiles, sept espèces d’oiseaux et 24 taxons de mammifères. Alors que les mammifères sont représentés par 13 taxons de micromammifères (sept appartiennent à l’ordre Rodentia, cinq à l’ordre Eulipothypla, et un à l’ordre Chiroptera), les grands mammifères sont représentés par onze espèces, y compris des ongulés et des carnivores. Cette association faunique suggère un paléoenvironnement caractérisé par de vastes étendues boisées avec quelques prairies et un cours d’eau à proximité, dans un climat particulièrement chaud et humide. Ces conditions paléoenvironnementales, combinées à des datations AMS et AAR à partir d’échantillons d’os de la macrofaune, permettent de localiser le gisement d’Artazu VII dans la première moitié du Pléistocène supérieur, et plus précisement dans le stade isotopique marin (MEI) 5c

Global genomic diversity of a major wildlife pathogen: Ranavirus, past and present

Ranavirus is a genus of large double-stranded DNA viruses (family Iridoviridae) that parasitise three taxonomic classes of poikilothermic vertebrates. They are important wildlife pathogens of conservation and economic concern, posing significant threat to amphibian biodiversity and aquaculture commerce. Despite substantial advances since their discovery in the 1960s, the evolutionary history of ranaviruses remains poorly characterised. The aim of this thesis is to advance the characterisation of Ranavirus evolutionary dynamics to contemporary standards. A large whole-genome dataset was collated and scrutinised, combining all publicly available material with a novel collection of isolate genomes. Cutting-edge microbial genomics tools were applied to gain insight into ranavirus genetic diversity, phylogeography, and genome evolution. Delineation of the Ranavirus pan-genome served as a foundation to conduct phylogenetic and population genetic analyses. Where the limitations of alignment-based methodologies were met, alignment-free techniques were employed to make full use of all genomic information. Phylogenetic reconstructions uncovered unique genetic diversity incompatible with current taxonomic demarcations amongst several lineages of Ranavirus. Pervasive genetic recombination was detected across the genus, and certain lineages contained a high degree of ancestral polyphyly. Recurrent patterns linked to animal trade and aquaculture were detected. Extensively polyphyletic viruses were isolated from captive animals, and population genetic analysis revealed ancestry components shared by ranaviruses isolated from farmed animals on separate continents. Finally, phylodynamic analysis suggests human-mediated translocation of FV3-like ranaviruses began more than a century before present. The inadequacies of current Ranavirus taxonomy are highlighted by this work, and suggests a substantial diversity remains to be characterised. The processes by which ranaviral genetic diversity is generated appears particularly dynamic, with significant contributions made via recombination between distinct linages. Altogether, this thesis underscores the vital impact trade and captive rearing of fish and herpetofauna have had on the global spread of ranaviruses and their processes of genetic diversification. Finally, these results suggest that anthropogenic influences commenced decades earlier than previously thought, likely upon the acceleration of modern globalisation