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Pigeons are considered to play key ecological roles in frugivory and seed dispersal. They have colonised numerous oceanic islands and diversified into several species in sympatry. How these species coexist in similar niches is poorly understood although dietary separation is among the mechanisms suggested to avoid trophic overlap. We investigated the trophic ecology of the two endemic Columba species co-occurring in the laurel forest and thermosclerophyllous relicts of two of the Canary Islands. This study includes diet description in spatiotemporal terms, its relationship with fruit availability and seed treatment in 10 study areas established on La Palma and La Gomera. We used non-invasive DNA analysis to identify the faeces of the two con-generic species and microhistological methods to examine their diets. The degree of trophic overlap was evaluated by niche similarity and breadth indices. Molecular faecal sampling determined the spatiotemporal distribution of both pigeons to identify their areas of coexistence. These frugivorous pigeons’ diets did not differ concerning the main plant species, but they diverged quantitatively in the proportions and parts of plants consumed. Lauraceae fruits were their staple foods although Rhamnaceae and some Fabaceae and Solanaceae were also important. Both pigeons showed selective preferences for some fruits. Significant spatiotemporal variations in their diets were observed along with a general tendency to increase fruit intake at its ripening times. Our results suggest that different trophic strategies facilitate the coexistence of these frugivorous columbids. These pigeons act as seed dispersers and/or predators depend ing on seed features (size and hardness), and this may have valuable implications for their conservation., This study was partially funded by the Organismo Autónomo de Parques Nacionales (80/2005), Ministerio de Medio Ambiente y Medio Rural y Marino of Spain. PM received grant support from Proyectos Intramurales Especiales (2004 3 0E 169) of the Spanish National Scientific Research Council. The manuscript was edited by Guido Jones, currently funded by the Cabildo de Tenerife, under the TFinnova programme supported by MEDI and FDCAN funds.

Humans have transformed most landscapes across the globe, forcing other species to adapt in order to persist in increasingly anthropogenic landscapes. Wide-ranging solitary species, such as wild felids, struggle particularly in such landscapes. Conservation planning and management for their long-term persistence critically depends on understanding what determine survival and what are the main mortality risks. We carried out the first study on annual survival and cause-specific mortality of the European wildcat with a large and unique dataset of 211 tracked individuals from 22 study areas across Europe. Furthermore, we tested the effect of environmental and human disturbance variables on the survival probability. Our results show that mortalities were mainly human-caused, with roadkill and poaching representing 57% and 22% of the total annual mortality, respectively. The annual survival probability of wildcat was 0.92 (95% CI = 0.87–0.98) for females and 0.84 (95% CI = 0.75–0.94) for males. Road density strongly impacted wildcat annual survival, whereby an increase in the road density of motorways and primary roads by 1 km/km2 in wildcat home-ranges increased mortality risk ninefold. Low-traffic roads, such as secondary and tertiary roads, did not significantly affect wildcat's annual survival. Our results deliver key input parameters for population viability analyses, provide planning-relevant information to maintain subcritical road densities in key wildcat habitats, and identify conditions under which wildcat-proof fences and wildlife crossing structures should be installed to decrease wildcat mortality., This research was funded by: the German Federal Ministry of Transport and Digital Infrastructure (BMVI) as part of the mFund project “WilDa—Dynamic Wildlife–Vehicle Collision warning, using heterogeneous traffic, accident and environmental data as well as big data concepts” grant number 19F2014B; the Deutscher Akademischer Austauschdienst (DAAD) Research Grants, Short-Term Grants, 2020 (57507441); the Deutsche Wildtier Stiftung (DeWiSt). The data from Cabañeros National Park were collected in the frame of the project OAPN 352/2011 funded by Organismo Autónomo Parques Nacionales. MM was supported by a research contract Ramón y Cajal from the MINECO (RYC-2015-19231). FDR was supported by a postdoctoral contract funded by the University of Málaga through the grants program “Ayudas para la Incorporación de Doctores del I Plan Propio de Investigación de la Universidad de Málaga (Call 2019)”. PM was supported by UIDB/50027/2020 with funding from FCT/MCTES through national funds.

Amphibians are the most highly threatened vertebrates, and emerging pathogens are a serious threat to their conservation. Amphibian chytrid fungi and the viruses of the Ranavirus genus are causing disease outbreaks worldwide, including in protected areas such as National Parks. However, we lack information about their effect over amphibian populations in the long-term, and sometimes these mortality episodes are considered as transient events without serious consequences over longer time-spans. Here, we relate the occurrence of both pathogens with the population trends of 24 amphibian populations at 15 sites across a national Park in northern Spain over a 14-year period. Just one out 24 populations presents a positive population trend being free of both pathogens, while seven populations exposed to one or two pathogens experienced strong declines during the study period. The rest of the study populations (16) remain stable, and these tend to be of species that are not susceptible to the pathogen present or are free of pathogens. Our study is consistent with infectious diseases playing an important role in dictating amphibian population trends and emphasizes the need to adopt measures to control these pathogens in nature. We highlight that sites housing species carrying Ranavirus seems to have experienced more severe population-level effects compared to those with the amphibian chytrid fungus, and that ranaviruses could be just as, or more important, other more high-profile amphibian emerging pathogens., This study was funded by the Spanish Ministry for Science, Innovation and Universities (CGL2015-70070-R, PI: JBo and RTI2018-095023-B-I00, PI: Luz Boyero) and by Organismo Autónomo Parques Nacionales of Spain (2399/2017, PI: JBo).We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

This study evaluates the potential for pasture improvement and restoration at two silvopastoral sites. We used a total of 16 wild legume species under different management systems (rhizobial inoculation, mycorrhizal inoculation, sheep penning, and sheep penning with mycorrhizal inoculation), at two mountain sites in Sierra Nevada Natural Park (Granada, southern Spain). The first site, Soportújar, had higher soil fertility and rainfall than the second, Lanjarón. Forage yields in Soportújar ranged from 265 to 8970 kg DM ha¿1, Vicia amphicarpa, Vicia monantha, Vicia disperma and Lathyrus clymenum being the most productive species. Mycorrhizal inoculation resulted in higher forage yields for Lens nigricans, V. disperma and Vicia lutea. Seed yields were low, ranging from 5.9 to 1234 kg ha¿1. Forage yields in Lanjarón were lower, ranging from 46 to 1415 kg DM ha¿1; and the most productive species were V. monantha, V. disperma, Lathyrus cicera and Medicago rigidula. Sheep penning alone and together with mycorrhizal treatment resulted in greater forage yields for most of the species studied, although differences were only significant for V. disperma, V. monantha and Lathyrus sphaericus. Seed yields ranged from 0.4 to 60 kg ha¿1. In conclusion, we recommend V. monantha, V. disperma and L. cicera followed by V. amphicarpa and L. clymenum (in wetter more fertile sites) and M. rigidula (in drier sites), as they seem to be the best adapted to the pedoclimatic conditions of Sierra Nevada Natural Park. Increasing nutrients in the soil (by sheep penning) and promoting nutrient assimilation (by mycorrhizal inoculation) may be effective strategies for increasing pasture biomass in silvopastoral sites., This work has been funded by the National Parks Autonomous Agency (OAPN, Spanish Ministry of Agriculture, Food and Environment) through the project “Investigaciones sobre la flora forrajera natural en mejoras de pastos, restauración forestal y silvicultura preventiva con ganado: una experiencia piloto en Sierra Nevada” (Ref. 748). We would like to thank Baltasar del Pozo for managing the sheep penning, Mycovitro who donated the Glomigel, and Manuel Fernández-López and Pablo Villadas who provided the rhizobium inocula for this study.

Large, long-lived species with slow life histories and protracted pre-breeding stages are particularly susceptible to declines and extinction, often for unknown causes. Here, we show how demographic modeling of a medium-sized raptor, the Red Kite Milvus milvus, can aid to refocus conservation research and attention on the most likely mechanisms driving its decline. Red Kites’ survival and reproduction increased through three sequential stages for 1–2, 3–6, and 7–30 yr of age, mainly corresponding to individuals that are dispersing, attempting to gain a territory, and breeding. As typical of long-lived species, elasticities were highest for adult (≥7 yr old) survival, but this was high, with little scope for improvement. Instead, the declines were driven by an extremely low survival of pre-adults in their first years of life, which weakened the whole demographic system by nullifying the offspring contribution of adults and curtailing their replacement by recruits. For example, 27 pairs were necessary to generate a single prime age adult. Simulation of management scenarios suggested that the decline could be halted most parsimoniously by increasing pre-adult survival to the mean levels recorded for other areas, while only the synergistic, simultaneous improvement of breeding success, adult and pre-adult survival could generate a recovery. We propose three actions to attain such goals through selective supplementary feeding of both breeding and non-breeding individuals, and through mortality improvement by GPS remote-sensing devices employed as surveillance monitoring tools. Our results show how improving demographic models by using real, local vital rates rather than “best guess” vital rates can dramatically improve model realism by refocusing attention on the actual stages and mortality causes in need of manipulation, thus building precious time and resources for conservation management. These results also highlight the frequent key role of pre-adult survival for the management of long-lived species, coherent with the idea of demographic systems as integrated chains only as strong as their weakest link., This study was funded by the Foundation Jaime González-Gordon and by the research projects 1602/2015 of the Spanish Ministry of Agriculture, Food and the Environment (Autonomous Organism of National Parks), PGC2018-095860-B-I00 of the Ministry of Science, Innovation and Universities with Feder Funds, and P18-FR-4239 of the Andalucía Autonomous Region (Consejería de Economía, Conocimiento, Empresas y Universidad).

[Background] Syngnathid fishes (Actinopterygii, Syngnathidae) are flagship species strongly associated with seaweed and seagrass habitats. Seahorses and pipefishes are highly vulnerable to anthropogenic and environmental disturbances, but most species are currently Data Deficient according to the IUCN (2019), requiring more biological and ecological research. This study provides the first insights into syngnathid populations in the two marine Spanish National Parks (PNIA—Atlantic- and PNAC—Mediterranean). Fishes were collected periodically, marked, morphologically identified, analysed for size, weight, sex and sexual maturity, and sampled for stable isotope and genetic identification. Due the scarcity of previous information, habitat characteristics were also assessed in PNIA., [Results] Syngnathid diversity and abundance were low, with two species identified in PNIA (Hippocampus guttulatus and Syngnathus acus) and four in PNAC (S. abaster, S. acus, S. typhle and Nerophis maculatus). Syngnathids from both National Parks (NP) differed isotopically, with much lower δ15N in PNAC than in PNIA. The dominant species were S. abaster in PNAC and S. acus in PNIA. Syngnathids preferred less exposed sites in macroalgal assemblages in PNIA and Cymodocea meadows in PNAC. The occurrence of very large specimens, the absence of small-medium sizes and the isotopic comparison with a nearby population suggest that the population of Syngnathus acus (the dominant syngnathid in PNIA) mainly comprised breeders that migrate seasonally. Mitochondrial cytochrome b sequence variants were detected for H. guttulatus, S. acus, and S. abaster, and a novel 16S rDNA haplotype was obtained in N. maculatus. Our data suggest the presence of a cryptic divergent mitochondrial lineage of Syngnathus abaster species in PNAC., [Conclusions] This is the first multidisciplinary approach to the study of syngnathids in Spanish marine NPs. Habitat preferences and population characteristics in both NPs differed. Further studies are needed to assess the occurrence of a species complex for S. abaster, discarding potential misidentifications of genus Syngnathus in PNAC, and evaluate migratory events in PNIA. We propose several preferential sites in both NPs for future monitoring of syngnathid populations and some recommendations for their conservation., Study funded by Proyecto Hippoparques (1541S/2015 and 1580S/2015; Organismo Autónomo de Parques Nacionales—OAPN, Ministerio de Agricultura, Alimentación y Medio Ambiente, Spain). Project 1580S/2015 was in charge of genetics.

13 Págs., Rear-edge tree populations forming the equatorward limit of distribution of temperate species are assumed to be more adapted to climate variability than central (core) populations. However, climate is expected to become more variable and the frequency of climate extremes is forecasted to increase. Climatic extreme events such as heat waves, dry spells and spring frosts could become more frequent, and negatively impact and jeopardize rear-edge stands. To evaluate these ideas, we analyzed the growth response of trees to successive spring frosts in a mixed forest, where two temperate deciduous species, Fagus sylvatica L. (European beech) and Quercus petraea (Matt.) Liebl. (sessile oak), both at their southernmost edge, coexist with the Mediterranean Quercus pyrenaica Willd. (Pyrenean oak). Growth reductions in spring-frost years ranked across species as F. sylvatica > Q. petraea > Q. pyrenaica. Leaf flushing occurred earlier in F. sylvatica and later in Q. pyrenaica, suggesting that leaf phenology was a strong determinant of spring frost damage and stem growth reduction. The frost impact depended on prior climate conditions, since warmer days prior to frost occurrence predisposed to frost damage. Autumn Normalized Difference Vegetation Index data showed delayed leaf senescence in spring-frost years and subsequent years as compared with pre-frost years. In the studied forest, the negative impact of spring frosts on Q. petraea and especially on F. sylvatica growth, was considerably higher than the impacts due to drought. The succession of four spring frosts in the last two decades determined a trend of decreasing resistance of radial growth to frosts in F. sylvatica. The increased frequency of spring frosts might prevent the expansion and persistence of F. sylvatica in this rear-edge Mediterranean population., The National Parks Autonomous Agency (2483S/2017, 2481S/2017); the Autonomous Community of Madrid (P2013/ MAE-2760); the Spanish Ministry of Economy, Industry and Competitiveness (AGL2016-76769-C2-1-R, RTI2018-096884-B-C31); and the Spanish Ministry of Education, Culture and Sports (FPU15/03533 to A.R-.C.).

Invasive species pose a major threat to global diversity, and once they are well established their eradication typically becomes unfeasible. However, certain natural mechanisms can increase the resistance of native communities to invaders and can be used to guide effective management policies. Both competition and herbivory have been identified as potential biotic resistance mechanisms that can limit plant invasiveness, but it is still under debate to what extent they might be effective against well-established invaders. Surprisingly, whereas biotic mechanisms are known to interact strongly, most studies to date have examined single biotic mechanisms separately, which likely influences our understanding of the strength and effectiveness of biotic resistance against invaders. Here we use long-term field data, benthic assemblage sampling, and exclusion experiments to assess the effect of native assemblage complexity and herbivory on the invasion dynamics of a successful invasive species, the alga Caulerpa cylindracea. A higher complexity of the native algal assemblage limited C. cylindracea invasion, probably through competition by canopy-forming and erect algae. Additionally, high herbivory pressure by the fish Sarpa salpa reduced C. cylindracea abundance by more than four times. However, long-term data of the invasion reflects that biotic resistance strength can vary across the invasion process and it is only where high assemblage complexity is concomitant with high herbivory pressure, that the most significant limitation is observed (synergistic effect). Overall, the findings reported in this study highlight that neglecting the interactions between biotic mechanisms during invasive processes and restricting the studied time scales may lead to underestimations of the true capacity of native assemblages to develop resistance to invaders., Funding for this project was obtained from the Spanish Ministry Project ANIMA (CGL2016-76341-R, MINECO/FEDER, UE), the European Union’s EMFS Program, AFRIMED (EASME/EMFF/2017/1.2.1.12/S4/01/SI2.789059), the Spanish Autonomous Organism of National Parks through the project BIGPARK (project 2017-2462), and the Padi Foundation to FT (CGA App: 5134). JS received the support of a fellowship from “La Caixa” Foundation (ID 100010434) with code (LCF/BQ/DE17/11600001).

The reproductive biology of two species of pipefish, Syngnathus typhle and S. abaster, was investigated in western Mediterranean coastal waters, through determination of sex-ratio and maturity state, and histological assessment of gonads, brood pouch and embryos of these little-known syngnathid species. A total of 22 S. typhle and 13 S. abaster individuals were collected by towing-net on shallow seagrass meadows (Posidonia oceanica and Cymodocea nodosa) at Balearic Islands (Spain). Overall sex-ratio (male : female) was 1.0 : 1.2 for S. typhle and 1.0 : 2.7 for S. abaster. Regarding maturity, all stages of females were sampled while only immature or brooding males were collected. Males with embryos in the pouch were found in summer for both species, and mature females of S. typhle were found in summer whereas mature females of S. abaster were found in winter and summer, suggesting that each species might have a different reproductive cycle. This study contributes to the knowledge on the reproductive biology of the two most abundant pipefish species in W-Mediterranean coastal waters, which might help for further restocking and conservation management strategies., This study was partly funded by Proyecto Hippoparques (1541S/2015 and 1580S/2015; Organismo Autónomo de Parques Nacionales Ministerio de Agricultura, Alimentación y Medio Ambiente, Spain).

Pipefish are a vulnerable and diverse group of ichthyofauna tightly associated with seagrass meadows, key habitats in shallow marine areas. Despite their charismatic role, the main ecological features, habitat, and diet of this group remain largely unknown. This study focuses on assessing pipefish habitat and feeding preferences, including different hosting seagrasses such as Posidonia oceanica and Cymodocea nodosa from the Balearic Islands, western Mediterranean. Four species (Syngnathus typhle, S. abaster, Nerophis ophidion, and N. maculatus) were found associated to different seagrasses. S. typhle and N. maculatus were more frequent in P. oceanica meadows, while S. abaster and N. ophidion in C. nodosa. Individuals of all species captured in P. oceanica were larger than those living in C. nodosa, suggesting a size-dependent habitat preference. Feeding preferences, however, were driven by prey availability and fish features, e.g., head/snout morphology. For the first time in the western Mediterranean, a thorough description of the diet and potential prey of this group was carried out. Epifaunal assemblages (potential prey) were dominated in both habitats by harpacticoid copepods and gammarid amphipods, and they were also the primary prey according to stomach contents of all species. These results can contribute to future pipefish conservation and management actions, such as targeting crucial habitat identification and designing culture and reintroduction protocols., This study was funded by the research Project “Singnátidos de los Parques Nacionales de las Islas Atlánticas y del Archipiélago de Cabrera (541S/2015)” awarded to CSIC by Organismo Autónomo Parques Nacionales (Ministerio de Agricultura, Alimentación y Medio Ambiente, Programa Estatal de I+D+i Orientada a los Retos de la Sociedad, Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016). Additional support was provided by the joint Associated Unit IMEDEA-LIMIA (Laboratorio de Investigaciones Marinas y Acuicultura, Dirección General de Pesca y Medio Marino, Govern Illes Balears).

Among the scincid lizards inhabiting the Mediterranean region, the Ocellated skink (Chalcides ocellatus) and the little-known and endangered Chafarinas' skink (Chalcides parallelus) co-occur in sympatry at the Chafarinas Islands (NW Africa). Although both species are present in the archipelago, C. ocellatus is spread in the three islands, but C. parallelus is present only in one island. What factors influence this distribution, and whether these two species show evidence for syntopy is unknown. In this study, we investigated spatial niche occupancy at the level of microhabitat of both skinks at the Chafarinas Islands. We tested for microhabitat differences between islands and then, we tested the null hypothesis of no differences between microhabitat used by each of these skink species and a set of locations chosen at random from the general habitat. Results showed that the distribution and abundance of available microhabitats were different among islands, thus each island has different spatial niches that may be important in distinguishing the areas occupied by these skinks. Results also revealed that C. ocellatus did not use available microhabitats at random, but preferentially selected particular microhabitats. However, C. parallelus used microhabitats according to their availability, although it showed some aspects of habitat selection similar to C. ocellatus. Our results showed that the inclusion of detailed microhabitat descriptors could be useful for understanding patterns of niche occupancy by these skinks species. Furthermore, to our knowledge this is the first study documenting the spatial niche selection of C. parallelus and provides novel ecological information that may be of major importance to develop management strategies that will help to conserve this endangered species., Financial support was provided by Organismo Autónomo de Parques Nacionales (Spain), GENA SL and the Portuguese Science and Technology Foundation (FCT) through Postdoctoral Grant SFRH/BPD/109182/2015 to E. Civantos, with additional financial support from the Spanish Ministerio de Ciencia, Innovación y Universidades project PGC2018-093592-B-I00 (MCIU/AEI/FEDER, UE) to J. Martín.

Mesorhizobium contains species widely known as nitrogen-fixing bacteria with legumes, but their ability to promote the growth of non-legumes has been poorly studied. Here, we analyzed the production of indole acetic acid (IAA), siderophores and the solubilization of phosphate and potassium in a collection of 24 strains belonging to different Mesorhizobium species. All these strains produce IAA, 46% solubilized potassium, 33% solubilize phosphate and 17% produce siderophores. The highest production of IAA was found in the strains Mesorhizobium ciceri CCANP14 and Mesorhizobium tamadayense CCANP122, which were also able to solubilize potassium. Moreover, the strain CCANP14 showed the maximum phosphate solubilization index, and the strain CCANP122 was able to produce siderophores. These two strains were able to produce cellulases and cellulose and to originate biofilms in abiotic surfaces and tomato root surface. Tomato seedlings responded positively to the inoculation with these two strains, showing significantly higher plant growth traits than uninoculated seedlings. This is the first report about the potential of different Mesorhizobium species to promote the growth of a vegetable. Considering their use as safe for humans, animals and plants, they are an environmentally friendly alternative to chemical fertilizers for non-legume crops in the framework of sustainable agriculture., This research was supported by the Ministerio de Medio Ambiente y Medio Rural y Marino, Organismo Autónomo de Parques Nacionales (Ref. 111/2010) to M.L.-B. and by the Strategic Research Programs for Units of Excellence from Junta de Castilla y León (CLU-2O18-04) to E.V. It was also funded by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), and by Portuguese funds through FCT—Fundação para a Ciência e a Tecnologia, in the framework of the project POCI-01-0145-FEDER-016810 (PTDC/AGR-PRO/2978/2014 and the Project UIDB/05183/2020) to E.M. E.M. acknowledges an FCT contract from the Individual Call to Scientific Employment Stimulus 2017 (CEECIND/00270/2017), which funds her position.

Several reproductive issues and the larval development of the ferruginous limpet, Patella ferruginea Gmelin, 1791, an endangered species endemic from the western Mediterranean Sea, were studied to fill gaps in the knowledge of its life cycle. Average diameter of mature oocytes was 141.83 μm and mean oocyte density in the ovary was 283,800 oocytes/gram. No significant correlations were found between both oocyte diameter or density and female shell length. Female fecundity (number of oocytes per gonad) was significantly correlated with shell length and varied between 189,200 oocytes in a 40.0 mm female and 5,019,200 oocytes in an 86.4 mm female. However, there was considerable variability, in particular for the largest females. Spawning induction was not achieved using usual molluscan aquaculture methods. Thus, oocytes obtained after dissection of females were used for fertilisations trials. Alkalinisation treatments of seawater were used to test improvement in oocyte maturation and later fertilisation rates. Treatment at pH 9 during 2 h produced the highest increase in the percentage of mature oocytes and in the fertilisation rate; but these results showed high variability and were mainly significant when the initial degree of maturation was low. Sperm concentration experiments determined that best in vitro fertilisation was performed at 10 and 5×10 spermatozoids/ml. The sequence and timing of the complete larval development of Patella ferruginea in laboratory conditions is described and illustrated here for the first time. At 20 °C, larvae became competent for metamorphosis 3 days after fertilisation, but some crawling pediveliger larvae with a still well-developed velum were found even 7 days after fertilisation. Recruits 1–2 mm in length were achieved in low numbers from two of the assays and were first detected between 131–141 days after fertilisation. The resulting juveniles were monitored for two years and sex determination of five survivors at the end of this period showed that two were mature males, two mature females and one indeterminate. Our results show that the main reproductive traits or larval development of P. ferruginea hardly differ from those of other non-endangered Mediterranean or NE Atlantic limpet species. Therefore, its decline cannot be mainly attributed to constraints of these traits as was previously suggested, but to human impact. On the other hand, it is feasible to complete the life cycle of this species in laboratory conditions, from fertilised eggs to mature individuals. However, an important part of the process like spawning induction was not achieved as gonads needed to be dissected fatally from females, although sperm could be obtained from males through non-lethal biopsies. At present, large-scale aquaculture production for reintroduction, restocking or stock enhancement purposes is neither possible nor an advisable conservation tool yet. Further study is required and meanwhile, the appropriate design of a network of effectively protected marine areas that ensures connectivity among extant populations is necessary., This work was funded at initial stages (2006-2011) by contracts made by the Organismo Autónomo de Parques Nacionales (OAPN) and the Dirección General de Conservación de la Naturaleza of the former Spanish Ministerio de Medio Ambiente (1996-2008), Ministerio de Medio Ambiente, y Medio Rural y Marino (2008-2011) and Ministerio de Agricultura, Alimentación y Medio Ambiente (2011-2016). At its late stage (2011-2012), was funded by the project “Action plan for viability proposals of the endangered limpet, Patella ferruginea” (Project Cero of the Spanish Research Council-CSIC Foundation).

Ivermectin is the most common endectocide used to control parasites affecting livestock. Short-term physiological and behavioural effects of ivermectin on dung beetles may have long-term consequences for beetle populations and ecosystem functioning. Long-term effects of the use of ivermectin can be estimated by comparing dung assemblages and ecosystem functions in areas with conventional ivermectin-treated livestock and environmentally similar areas in which livestock are not treated with veterinary medical products (organic farming). In this study, we investigated both short-term and long-term effects of the administration of ivermectin on the characteristics of dung beetle assemblages and the services they provided in a protected area (Doñana National Park, SW Spain). We examined short-term dung colonization, dwelling, relocation, and disaggregation rates and the associations between these processes and the key assemblage parameters of species richness, abundance, biomass and functional diversity. Furthermore, we analysed changes in soil physical-chemical properties and processes. Short-term differences were observed in the total amount of dung relocated by dung beetles at different colonization vs. emigration stages, suggesting that dung beetles in this area were affected by the recent treatments of livestock with ivermectin. Moreover, short-term effects could also be responsible for the significant differences in dung spreading rates between sites. Conventional use of ivermectin disrupted ecosystem functioning by affecting species richness, abundance and biomass. The decrease in diversity parameters was related to a reduction in the functional efficiency, which resulted in the long-term accumulation of dung on the ground and considerable changes in soil functionality., Financial support was provided by the projects CGL2015-68207-R of the Secretaría de Estado de Investigación– Ministerio de Economía y Competitividad, and OAPN 762/2012 of the Organismo Autónomo de Parques Nacionales-Ministerio de Agricultura, Alimentación y Medio Ambiente.

In nature, animals are exposed to a broad range of threats imposed by predators, which may strongly influence the ecology of prey species directly or indirectly by affecting their behavior via fear of predation. Here, we studied wood mice Apodemus sylvaticus behavioral and physiological responses to simulated predation risk. Risk avoidance was analyzed by live trapping with control traps and traps treated with feces of common genet Genetta genetta (direct cue of risk) under new moon nights and following by simulated full moon conditions (indirect cue). The time devoted to foraging behavior and capture time were analyzed by video recording mice activity around traps. Food intake was calculated based on the amount of bait remaining in each trap. Fecal corticosterone metabolites (FCMs) were measured by enzyme-immunoassay as indicators of physiological stress responses. Fewer wood mice were captured during full moon, yet only non-breeding adult males clearly avoided common genet odor. Mice were captured sooner at night during the simulated full moon conditions and later in predator-treated traps. Foraging activity was lower when individuals faced predator’s feces, but neither food intake nor FCM levels were affected by predation risk cues. Direct and indirect cues of predation risk selectively affected wood mice behavior, although behavioral responses seem to be modulated by different costs–benefit balances related to the individual’s perception of risk. The lack of physiological responses to predation risk cues suggests that wood mice did not perceive them as reliable stressors or the response was too small or transient to be measured by FCM., This work was partially supported by the project RISKDISP (CGL2009-08430/BOS) and Comunidad de Madrid together with the European Social Fund and Universidad Autónoma de Madrid (CCG10-UAM/AMB-5325). This study is a contribution to the projects MONTES [CSD2008-00040], RISKDISP [CGL2009-08430], and VULGLO [CGL2010-22180-C03-03], funded by the Spanish Ministerio de Ciencia e Innovación; 096/2002 and 003/2007, funded by the Organismo Autónomo Parques Nacionales; and ANASINQUE [PGC2010-RNM-5782], funded by the Junta de Andalucía. Á.N.-C. was supported by a FPU scholarship [AP2008-03430] from the Ministerio de Educación y Ciencia of Spain

The Tablas de Daimiel National Park (TDNP) is a unique floodplain ecosystem in central Spain, serving as permanent resting and breeding areas for many waterbird species. In the last decades, this biodiversity hotspot has been severely endangered by poorly treated wastewater discharges from upstream urban communities arriving through its two major contributors, the Cigüela and Guadiana rivers. In this work, we analysed the potential risk of this constant input of micropollutants (estrogens, dioxin-like compounds and other endocrine disruptors) for the resident wildlife. We sampled 12 locations in TDNP and in the nearby Navaseca Pond during 2013, and performed a series of in-vivo and in-vitro bioassays, including Daphnia magna post-exposure feeding inhibition and recombinant yeast-based assays for dioxin-like and estrogenic activities. These results were then compared with the chemical composition of the samples, analysed by GC–MS/MS and LC-MS/MS, and evaluated according to their toxic potential as toxic equivalents or TEQ. The Navaseca Pond, heavily impacted by wastewater from the town of Daimiel, showed the highest levels of toxic compounds, estrogenic activity, and Daphnia toxicity. Conversely, the less impacted TDNP sites showed low residue levels of contaminants, low estrogenicity and dioxin-like activity and negligible toxicity. The results indicates that the current good chemical status of TDNP is menaced by both the inflow of wastewater treatment plants effluents from Guadiana and Cigüela rivers into TDNP tributaries and, as it occurs in the Navaseca Pond, by direct sewage discharges., This study was supported by the Spanish grants (OAPN755/2012 and CTM2014-51985). Claudia Rivetti was supported by a Spanish fellowship FPI BES-2012-053631 from the Ministry of Economy and Competitively, Celia Laguna was supported by a research contract funded by EU FEDER through the Regional Government of Castilla-La Mancha (PEIC2014-005-P), and Jhon J. López - Perea was supported by a doctoral FPU grant (AP-2012-04966) from the Spanish Ministry of Education.

Crimean–Congo haemorrhagic fever virus (CCHFV) is the causative agent of the severe tick‐borne, often fatal, zoonotic Crimean–Congo haemorrhagic fever (CCHF), which is widely distributed worldwide. The CCHFV transmission to humans occurs through tick bite, crushing of engorged ticks or contact with infected host blood. Previously, CCHFV genotype Africa III was reported in Spain. Given the emergence of CCHF and the role of ticks in pathogen maintenance and transmission, we investigated the presence and genotype identity of the virus in tick species parasitizing abundant wild host species in south‐western Spain. A total of 613 adult ticks were collected from hunter‐harvested wild ungulates in twenty locations throughout south‐western Spain. Ticks were identified, nucleic acids were extracted, RNA was analysed by a nested RT‐PCR targeting CCHFV S segment, and the amplicons were sequenced. According to the 212‐bp sequence amplified, the presence of CCHFV human genotype Europe V was detected in Hyalomma lusitanicum and Dermacentor marginatus ticks collected from red deer, fallow deer and Eurasian wild boar in different locations from south‐western Spain. Genotype Africa IV was also detected in a H. lusitanicum tick collected from a red deer. The detection of CCHFV in different tick species collected from various wild hosts and localities provided strong evidence of widespread CCHFV presence in the region, suggesting that the circulation of the virus in Spain requires more attention. Additionally, the identification of the CCHFV genotype Europe V in ticks suggested that its introduction in Spain was probably from Eastern Europe., This study was partially supported by MINECO Plan Nacional grant CGL2017‐ 89866‐R and EU‐FEDER and by the Health Sciences Institute of the Government of Castilla‐La Mancha (JCCM), Spain, through project BPLY/19/180501/000321. We acknowledge the contribution of “Organismo Autónomo de Parques Nacionales (OAPN)‐Centro Quintos de Mora” and "La Morera Estate" to the tick survey. AMF was funded by JCCM through contract SBPLY/18/180501/000009, and IGFM was supported by the Research Plan of the University of Castilla‐La Mancha (UCLM), Spain.

The Tablas de Daimiel National Park (TDNP) is a floodplain ecosystem in central Spain with a potential risk of heavy metal and metalloid pollution. The objective of this study was to know the accumulation of arsenic (As), cadmium (Cd), mercury (Hg), lead (Pb), and selenium (Se) in muscle of six species of freshwater fish from the floodplain of TDNP. We obtained muscle samples of Cyprinus carpio (n = 89); Squalius pyrenaicus (n = 16); Ameiurus melas (n = 9); Lepomis gibbosus (n = 8); Micropterus salmoides (n = 6) and Carassius auratus (n = 5). A. melas, which is a predatory species, had significantly higher Hg concentrations than omnivorous or herbivorous species (i.e. C. carpio). On the contrary, A. melas showed lower concentrations of As, Pb and Se than omnivorous species (i.e. S. pyrenaicus and L. gibbosus ). The concentration of Hg was positively associated with fish size in C. carpio and A. melas. Some individuals of C. carpio (5.7%) and S. pyrenaicus (12.5%) showed Pb muscle concentrations above the maximum residue levels established by the European legislation for human consumers. The observed muscle Se concentrations can be associated with adverse effects on fish such as blood changes, reduced growth, mortality of juveniles and reproductive failure. The accumulation of Se in this floodplain located in a seleniferous area and the contamination produced by spent Pb shot pellets used for hunting in the past are discussed as potential sources of the elevated levels of these two elements in fish from this floodplain wetland., This study was supported by grant of research in National Parks (OAPN 755/2012) from the Ministry of Agriculture, Food and Environment Spain. Jhon J. López-Perea was supported by a doctoral FPU grant (AP-2012-04966) from the Spanish Ministry of Education. Sergio Fernández-Trujillo also thanks University of Castilla-La Mancha for the research starting scholarship (BIN1611), and Junta de Comunidades de Castilla-La Mancha for his pre-doctoral contract, SBPLY/16/180501/000356.

[Aim] To obtain nuclear and mitochondrial phylogenies of Macaronesian Rhopalomesites weevils and test the monophyly and time of origin of two species groups feeding on Euphorbiaceae and other plants. Additionally, to investigate the population structure within species, and its associations with geographical isolation versus trophic selection. [Location] Macaronesian Islands and the British Isles. [Methods] Maximum likelihood and Bayesian phylogenetic analyses were undertaken using mitochondrial (cox1 and cytb) and nuclear (ITS-2 and 28S RNA) genomic sequences. Ancestral Rhopalomesites host plant associations and divergence times were inferred from Bayesian analyses and population data. [Results] Evidence was found for two Rhopalomesites clades. One was associated with Euphorbia host plants, having vicariant species in the Madeira and Canary archipelagos. In this lineage, an ancestral association with Euphorbia mellifera in the two island groups was deduced, which has subsequently undergone shifts to related host plant species in marginal areas. The second clade was found to be an ecological generalist, exploiting decaying wood from the Lauraceae or other forest trees – is also present on such islands along with the Azores and parts of Atlantic Western Europe. [Main conclusions] The results point to a quasi-parallel colonization by the two ecologically distinct lineages in Macaronesia, dating to the early Pliocene, followed by allopatric isolation according to the presence of suitable habitats on particular islands in more recent times., This study was financed by a Spanish MMA grant (Organismo Autónomo de Parques Nacionales, 025/2007), and partially by a Spanish MCINN grant (CGL2009-08256) and a Netbiome Project (ISLAND-BIODIV) of the European Fund for Regional Development and the Canary Agency for Research. David Hernández-Teixidor held a PhD grant awarded by the Canary Government, 85% financed from the European Social fund, and then by a Spanish FPU fellowship. We appreciate the funding provided to the ULL by the Agencia Canaria de Investigación, Innovación y Sociedad de la Información, financed 85% by the European Social Fund.

[Context]: Camera trapping is commonly employed for studying carnivores because it provides better data than do other methods, and with lower costs. Increasing the probability of detecting the target species can reduce parameter uncertainty and survey effort required to estimate density and occurrence. Different methods, including attractants and baits, can be used to increase detectability. However, their efficacy has rarely been quantitatively assessed. [Aims]: To quantify the efficacy of scent and bait attractants to increase the detection probability of mesocarnivores by camera trapping. [Methods]: We tested two scent lures, valerian extract (Val) and lynx urine (LU), their combination (Val-LU), and a non-reward bait (i.e. not accessible to the animal), raw chicken, as carnivore attractants in two protected areas of central Spain. We used camera-trapping records under an occupancy-modelling framework to estimate attractant-specific detection probabilities for six mesocarnivore species, and quantified their effects by comparing these estimates with the baseline (i.e. no attractant) detectability. [Key results]: The weekly detection probability of four mesocarnivore species (red fox, Vulpes vulpes, stone marten, Martes foina, common genet, Genetta genetta, and Eurasian badger, Meles meles) significantly increased when Val-LU (between 0.11±0.07 and 0.67±0.05) or chicken (between 0.31±0.02 and 0.77±0.22) were used as attractant, when compared with baseline detectability (between 0.01±0.02 and 0.29±0.05). Although rarely, wildcats (Felis silvestris) were mostly detected with Val-LU. The probability of detecting a species after k sampling occasions (7 days each) where it was present was highly improved with both Val-LU and chicken for all species (e.g. from 0.20 to 0.98 after four sampling occasions with chicken for the stone marten). Both attractants reduced the sampling time required to ascertain that a species was absent to between 42% and 15% of baseline values. [Conclusions]: The tested attractants greatly improved the detectability of most Iberian mesocarnivores. Although chicken was preferable for some species such as stone marten, Val-LU is most efficient for detecting the whole mesocarnivore community, including rare species such as wildcats. [Implications]: Attractant selection in studies using non-invasive detection methods must be supported by quantitative assessment of the detection probability associated to each attractant. Researchers must choose those attractants best fitting target species and study aims., This study was funded by Organismo Autónomo Parques Nacionales, project OAPN 352/2011. F. Díaz-Ruiz was supported by a Juan de laCierva postdoctoral contract (Ministerio de Ciencia e Innovación, Funder Id: 10.13039/501100004837, FJCI-2015-24949) from the Spanish Ministry of Economy, Industry and Competitiveness). P. Monterroso was granted a postdoctoral fellowship by the project ‘Genomics Applied to Genetic Resources’ co-financed by North Portugal Regional Operational Program 2007/2013 (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF).

The Eurasianwild boar has experienced aworldwide demographic explosion that increases awareness on shared pathogens. However, shedding routes of relevant wild boar pathogens are unknown. Previous observations on sex- and age-related differences in Aujeszky's disease virus (ADV) exposure led us to hypothesize that shedding patterns of endemicwild boar pathogens may be influenced by individual traits.We investigated shedding routes of ADV, porcine parvovirus (PPV), porcine circovirus type 2 (PCV2) and Coxiella burnetii and analysed the effect of host sex and age on pathogen shedding patterns. The presence of pathogen antibodies in serumand of pathogen DNA in oral, nasal, genital and rectal swabswas analysed by ELISA and PCR, respectively. The influence of sex and age in pathogen shedding prevalencewas tested statistically.Main routes of ADV, PPV, PCV2 and C. burnetii shedding were identified but the hypothesis of sex- and/or age-related shedding patterns couldn't be confirmed., This study is a contribution to European Commission grants ANTIGONE (278976) and APHAEA (EMIDA ERA-NET) and to agreement between OAPN and IREC for ‘Quintos de Mora’ estate. FRF acknowledges funding from ‘Juan de la Cierva’ and ‘Ramón y Cajal’ contracts (Spanish Ministry for the Economy and Competitiveness – MINECO).

[Aim]: A striking case of mitochondrial DNA (mtDNA) introgression occurs in the Iberian Peninsula: the mtDNA of Lepus timidus, a species that inhabited this territory during the Last Glacial Maximum (LGM), is almost fixed in the Iberian range of Lepus europaeus, is very frequent in the northern half of Iberia within Lepus granatensis, and is fixed in Lepus castroviejoi. Here, we used a transdisciplinary approach to understand the ecological drivers of the distribution of this introgressed mtDNA in the Iberian hare species. [Location]: Europe, with an emphasis on the Iberian Peninsula. [Methods]: First, we modelled the climatic niche of L. timidus to predict the favourability for the species during the LGM. Second, we gathered data regarding L. timidus mtDNA introgression for 1137 individuals (139 populations) from the Iberian Peninsula. We tested whether the climatic favourability for L. timidus during the LGM reflects the frequency of the introgressed mtDNA in the carrier hare populations. [Results]: The climatic favourability for the LGM is positively related to the presence of individuals with L. timidus mtDNA, and discriminates better than by chance between individuals with and without this lineage. At the population level, the favourability values relate to the observed frequencies of mtDNA introgression and discriminate between populations with and without L. timidus mtDNA. [Main conclusions]: The results suggest that the geographical distribution of L. timidus mtDNA introgression is linked to L. timidus distribution in Iberia during the LGM. Our study helps to improve our understanding of the role of climate in the reticulated evolutionary history of hares, and is a first step towards understanding the ecological impact of mtDNA introgression on Iberian hares., P.A. is currently supported by the Spanish Ministerio de Economía y Competitividad (MINECO) and Universidad de Castilla-La Mancha (UCLM) through a ‘Ramón y Cajal’ contract (RYC-2012-11970). J.M.-F. and R.C. hold FCT Investigator and postdoctoral grants, respectively, from the Fundaçao para a Ciência e a Tecnologia (FCT), funded by Programa Operacional Potencial Humano (POPH) – Quadro de Referencia Estrategico Nacional (QREN) from the European Social Fund and by the Portuguese Ministerio da Educacao e Ciencia (IF/00033/2014 and SFRH/BPD/64365/2009, respectively). This work is funded by FEDER funds through the Operational Programme for Competitiveness Factors – COMPETE and by National Funds through FCT – Foundation for Science and Technology under the EXPL/AAG-MAA/1082/2013 and FCOMP-01-0124-FEDER-041082. Partial funding was also obtained from FCT-ANR/BIA-EVF/0250/2012 and by the Spanish Ministry of Agriculture, Food and Environment, Spanish National Park’s Network (project 1098/2014).

Altitude significantly affects vegetation growth and distribution, including the developmental stages of a forest. We used shotgun Illumina sequencing to analyze microbial community composition and functional potential in melojo-oak (Quercus pyrenaica Willd.) rhizospheric soil for three different development stages along an altitudinal gradient: (a) a low altitude, non-optimal site for forest maintenance; (b) an intermediate altitude, optimal site for a forest; and (c) a high altitude, expansion site with isolated trees but without a real forest canopy. We observed that, at each altitude, the same microbial taxa appear both in the taxonomic analysis of the whole metagenome and in the functional analysis of the methane, sulfur and nitrogen metabolisms. Although there were no major differences at the functional level, there were significant differences in the abundance of each taxon at the phylogenetic level between the rhizospheres of the forest (low and intermediate altitudes) and the expansion site. Proteobacteria and Actinobacteria were the most differentially abundant phyla in forest soils compared to the expansion site rhizosphere. Moreover, Verrucomicrobia, Bacteroidetes and Nitrospirae phyla were more highly represented in the non-forest rhizosphere. Our study suggests that rhizospheric microbial communities of the same tree species may be affected by development stage and forest canopy cover via changes in soil pH and the C/N ratio., This work was supported by research grants including ERDF (European Regional Development Funds): P08-CVI-03549 from Consejería de Innovación, Ciencia y Empresa from Junta de Andalucía, OAPN 021/2007 from Organismo Autónomo Parques Nacionales (Spanish Ministry of Environment) and 20134R069-RECUPERA 2020 from the Spanish Ministerio de Economía y Competitividad and CSIC. We would like to thank the authorities of the Sierra Nevada National Park for the access, facilities and soil sampling, and also Edward Kirton (JGI, California) for technical assistance with bioinformatics. JFCD was awarded a postdoctoral contract from Junta de Andalucía, and AJFG was awarded a postdoctoral contract from RECUPERA 2020. The work of the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User Facility, is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231.

There is a risk of ‘ecological surprises’ if multiple potentially interacting stressors are managed individually, which is a question attracting significant current interest. Habitat degradation and species introductions are major threats to global biodiversity, and riverine fish are among the most threatened taxa in the world. Our interest was whether the presence of non-native species can affect native fish sensitivity to water quality deterioration in a large region in northeastern Spain (99,700 km, 15 catchments, 530 sites). We used a ‘base model’ with geographical and hydro-morphological variables, which are the major shaping factors in rivers. We tested whether water pollution, non-native species, or their interaction provided an improved understanding of patterns of distributions and health measurements of the twelve most common native species. There was little evidence that variation in native species abundance, where they occurred, the presence of diseases and changes in mean fish length or body condition was affected by water deterioration, the presence of non-native species, or their interaction. The disease rate and occurrence of native species might be affected, to a minor degree, by water quality changes and the presence of non-native species. Environmental conditions between sites with and without non-native fish differed in the condition of riparian areas and in water quality. Based on presence-absence data and changes in abundances through weighted average equations we also derived potential safe levels of salinization, nutrient pollution, and pH for the native fish. Overall, additive effects of stressors prevail over interactions, and the restoration of natural hydro-morphology in rivers is likely to be the most effective management approach to improving the prospects for the native fish fauna., Field surveys were funded by the ‘Agència Catalana de l'Aigua (ACA)’, ‘Confederación Hidrográfica del Ebro (CHE)’, the Natural Parks of ‘Collserola’ and ‘Sant Llorenç del Munt i Serra de l'Obac’, the Sabadell Council, and the project FURIMED-2 (CGL2008-03388BOS). The manuscript elaboration was supported by the Severo Ochoa Program for Centres of Excellence in R + D + I (SEV-2012-0262) and a fellowship from the ‘Fundació Barcelona Zoo and Ajuntament de Barcelona’ awarded to AMV.

20 páginas, 1 figura, 3 tablas., Recent studies have reported an unusually high frequency of nonamplifying alleles at microsatellite loci in bivalves. Null alleles have been associated with heterozygous deficits in many studies. While several studies have tested for its presence using different analytical tools, few have empirically tested for its consequences in estimating population structure and differentiation. We characterised 16 newly developed microsatellite loci and show that null alleles are ubiquitous in the wedge clam, Donax trunculus. We carried out several tests to demonstrate that the large heterozygous deficits observed in the newly characterised loci were most likely due to null alleles.Wetested the robustness of microsatellite genotyping for population assignment by showing that well-recognised biogeographic regions of the south Atlantic and south Mediterranean coast of Spain harbour genetically different populations., CR was supported by the Spanish Ministry of Education (Grant no PR2010-0601) and the Spanish Scientific Research Council (CSIC). This research was financially supported by Organismo Autonomo de Parque Nacionales Grant OAPN122_2010 to CR, EM, JAC and PD. ADM was supported by the Strategic Research Themes of the University of the South Pacific.

Understanding how different ecological and evolutionary processes influence the distribution of pathogens within the environment is important from many perspectives including wildlife epidemiology, evolutionary ecology and conservation. The simultaneous use of ecological and evolutionary frameworks can enhance our conceptual understanding of host-parasite interactions, however such studies are rare in the wild. Using samples from 12 bird species caught across all habitats existing on an oceanic island, we evaluated how environmental variables, parasite host specificity and parasite phylogenetic relationships determine the distribution and prevalence of haemosporidians (Haemoproteus, Plasmodium and Leucocytozoon) in the wild living avifauna. Differences were found in the prevalence of Plasmodium, but not Leucocytozoon, strains between habitats. The warmest temperature best predicted Plasmodium prevalence in the low altitude habitats, which had the highest incidence of Plasmodium. The prevalence of Leucocytozoon lineages was associated with natural factors, i.e. rainfall, temperature and habitat, but the two most important predictors (from model averaging) for models of Leucocytozoon were anthropogenic: poultry farms and distance to a water reservoir. We found no relationship between local (Tenerife, Canary Islands) versus global host range indices (which assess the diversity of hosts that a parasite is observed to infect), thus global generalist lineages do not behave in the same way on Tenerife (i.e. they infected less avian hosts than was expected). Phylogenetic analysis revealed that the most abundant haemosporidians on Tenerife grouped with lineages found in African host species. Our data indicate that climatic and anthropogenic factors, plus proximity to the African mainland, are the main factors influencing the presence and distribution of avian haemosporidians on Tenerife. Future climate projections for the archipelago foresee significant temperature increases which would, given our results, increase rates of Plasmodium infection in bird species in all habitats. Such patterns could be of concern if those increase mortality rates in the unique avifauna of these islands., DPP was supported by a postdoctoral fellowship (based at the University of East Anglia (UEA), UK) from the Spanish Ministry of Economy and Competitiveness. CGQ was funded by a PhD grant from Colciencias, Colombia and by the School of Biological Sciences at UEA. JCI was supported by the Spanish Administration of National Parks, Spanish Minister of Agriculture, Food and Environment (grant ref.: 375/2011).

Obtaining fundamental biodiversity metrics such as alpha, beta and gamma diversity for arthropods is often complicated by a lack of prior taxonomic information and/or taxonomic expertise, which can result in unreliable morphologically based estimates. We provide a set of standardized ecological and molecular sampling protocols that can be employed by researchers whose taxonomic skills may be limited, and where there may be a lack of robust a priori information regarding the regional pool of species. These protocols combine mass sampling of arthropods, classification of samples into parataxonomic units (PUs) and selective sampling of individuals for mtDNA sequencing to infer biological species. We sampled ten lowland rainforest plots located on the volcanic oceanic island of Réunion (Mascarene archipelago) for spiders, a group with limited taxonomic and distributional data for this region. We classified adults and juveniles into PUs and then demonstrated the reconciliation of these units with presumed biological species using mtDNA sequence data, ecological data and distributional data. Because our species assignment protocol is not reliant upon prior taxonomic information, or taxonomic expertise, it minimizes the problem of the Linnean shortfall to yield diversity estimates that can be directly compared across independent studies. Field sampling can be extended to other arthropod groups and habitats by adapting our field sampling protocol accordingly., This research was supported by the ERA‐Net Net‐Biome research framework, financed through Canary Island Government ACIISI grants SE‐12/02 (PO) and SE‐12/04 (BE), cofinanced by FEDER; Portuguese FCT‐NETBIOME grant 0003/2011 (PB); French ANR‐NETBIOME grant n°11‐EBIM‐001‐01 (CT); Région Réunion council for research activities (DS); Université de La Réunion contract DGADD/PE/20120585 (DS). BCE was also supported by Spanish MINECO grant CGL2013‐42589‐P, cofinanced by FEDER, and Spanish MAGRAMA grant S20141203_002597 from the Organismo Autónomo Parques Nacionales.

Species whose geographical distribution encompasses both mainland and island populations provide ideal systems for examining potential isolation and genetic divergence. This has also interest from a conservationist point of view, as it is important to protect “evolutionarily significant units”. We report a phylogenetic mitochondrial DNA analysis comparing the populations of the three Chalcides skink species from the Chafarinas Islands (NW Africa) with specimens of the same species from the nearest mainland. We tested for the potential genetic distinctiveness of the skink island populations. However, the results of the comparison of the genetic variability of the mitochondrial coding gene cytb were conclusive showing that the genetic divergence between continental and island Chalcides species was either non-existent or extremely low. We discuss how genetic divergence may be lower than expected if separation time of the islands with the mainland has not been long enough or if the island skink populations were currently communicated via ocean rafting with individuals coming from the mainland ones., R. García-Roa benefited from a FPI grant, J. Ortega from a CSIC JAE-pre grant. Legal authorization and support for the study was provided by the Organismo Autónomo de Parques Nacionales (Spain), with additional financial support from the Ministerio de Economía e Innovación research project CGL2014-53523-P.

Using a series of standardized sampling plots within forest ecosystems in remote oceanic islands, we reveal fundamental differences between the structuring of aboveground and belowground arthropod biodiversity that are likely due to large-scale species introductions by humans. Species of beetle and spider were sampled almost exclusively from single islands, while soil-dwelling Collembola exhibited more than tenfold higher species sharing among islands. Comparison of Collembola mitochondrial metagenomic data to a database of more than 80 000 Collembola barcode sequences revealed almost 30% of sampled island species are genetically identical, or near identical, to individuals sampled from often very distant geographic regions of the world. Patterns of mtDNA relatedness among Collembola implicate human-mediated species introductions, with minimum estimates for the proportion of introduced species on the sampled islands ranging from 45% to 88%. Our results call for more attention to soil mesofauna to understand the global extent and ecological consequences of species introductions., This research was supported by the ERA‐Net Net‐Biome research framework, financed through: Canary Island Government ACIISI grants SE‐12/02 (PO), SE‐12/03 (JCC), SE‐12/04 (BE), cofinanced by FEDER; Portuguese FCT‐NETBIOME grant 0003/2011 (PB); French ANR‐NETBIOME grant no 11‐EBIM‐001‐01 (CT); Région Réunion council for research activities (DS), Université de La Réunion contract DGADD/PE/20120585 (DS). B.C.E. was also supported by Spanish MINECO grant CGL2013‐42589‐P, co‐financed by FEDER, and Spanish MAGRAMA grant S20141203_002597 from the Organismo Autonomo Parques Nacionales.

Understanding the importance of host genetic diversity for coping with parasites and infectious diseases is a long-standing goal in evolutionary biology. Here, we study the association between probability of infection by avian malaria (Plasmodium relictum) and individual genetic diversity in three blue tit (Cyanistes caeruleus) populations that strongly differ in prevalence of this parasite. For this purpose, we screened avian malaria infections and genotyped 789 blue tits across 26 microsatellite markers. We used two different arrays of markers: 14 loci classified as neutral and 12 loci classified as putatively functional. We found a significant relationship between probability of infection and host genetic diversity estimated at the subset of neutral markers that was not explained by strong local effects and did not differ among the studied populations. This relationship was not linear, and probability of infection increased up to values of homozygosity by locus (HL) around 0.15, reached a plateau at values of HL from 0.15 to 0.40 and finally declined among a small proportion of highly homozygous individuals (HL > 0.4). We did not find evidence for significant identity disequilibrium, which may have resulted from a low variance of inbreeding in the study populations and/or the small power of our set of markers to detect it. A combination of subtle positive and negative local effects and/or a saturation threshold in the association between probability of infection and host genetic diversity in combination with increased resistance to parasites in highly homozygous individuals may explain the observed negative quadratic relationship. Overall, our study highlights that parasites play an important role in shaping host genetic variation and suggests that the use of large sets of neutral markers may be more appropriate for the study of heterozygosity–fitness correlations., This study was funded by grants 69/2003 (Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia), CGL2010-21933-C02-01 (Ministerio de Ciencia e Innovación) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund). ESF was supported by a predoctoral scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund, and VGN enjoyed a FPI predoctoral scholarship from Ministerio de Ciencia e Innovación-EuropeanSocial Fund. J.O was supported by ‘Juan de la Cierva’ and ‘Severo Ochoa-EBD’ post-doctoral fellowships (Ministerio de Ciencia e Innovación).

10 páginas, 2 figuras, 5 tablas., In a resource management perspective, the understanding of the relative influence of the physical factors on species connectivity remains a major challenge and is also of great ecological and conservation biology interest. Despite the overfishing threat on the wedge clam Donax trunculus in Europe, relatively little information is known about its population genetic structure and connectivity and their consequences on conservation policies. We employed 16 microsatellite loci to characterise the genetic diversity and population structure of D. trunculus. A total of 514 samples from seven different localities along the Atlantic-Mediterranean transition, from the Atlantic (Gulf of Cádiz) to the north-western Mediterranean were genotyped. The analysis of the population genetic structure displayed a clear distinction along the Atlantic-Mediterranean transition with different clusters in the Atlantic Ocean, the Alboran Sea and the northwestern Mediterranean. Consequently, we recommend that these three areas should be considered as different management units. We showed that all populations seem to be at high long-term risk of extinction with the exception of the protected Doñana National Park population which still seems to have evolutionary potential. Therefore, our results emphasized the necessity of protection of this economic resource and the validity of molecular tools to evaluate the population dynamics., This research was financially supported by Organismo Autónomo de Parques Nacionales Grant OAPN122_2010 to C.R., E.M., J.A.C. and P.D.

Treeline ecotones are considered early-warning monitors of the effects of climate warming on terrestrial ecosystems, but it is still unclear how tree growth at treeline will track the forecasted temperature rise in these cold environments. Here, we address this issue by analysing and projecting growth responses to climate on two different cold-limited alpine treelines: Pinus uncinata Ram. in the Spanish Pyrenees and Larix sibirica Ledeb. in the Russian Polar Urals. We assess radial-growth changes as a function of tree age and long-term climate variability using dendrochronology and a process-based model of tree growth. Climate‒growth relationships were compared considering young (age &lt, 50 years) and old trees (age &gt, 75 years) separately. Warm summer conditions enhanced radial growth, particularly after the 1980s, in the Polar Urals sites, whereas growth was positively related to warm spring and winter conditions in the Pyrenees sites. These associations were stronger in young than in old trees for both tree species and regions. Forecasted warm conditions are expected to enhance growth rates in both regions, while the growing season is forecasted to lengthen in the Pyrenees treelines, mostly in young trees. The observed age-related responses to temperature also depend on the forecasted warming rates. Although the temperature sensitivity is overall increasing for young trees, those responses seem more divergent, or even reversed, throughout the contrasting emission scenarios. The RCP 8.5 emission scenario corresponding to the most pronounced warming and drier conditions (+4.8 &deg, C) could also amplify drought stress in young trees from the Pyrenees treelines. Our modelling approach provides accessible tools to evaluate functional thresholds for tree growth in treeline ecotones under warmer conditions.

Direct observations of sex change were made on the endangered limpet Patella ferruginea in Chafarinas Islands (Alboran Sea) between 2006 and 2011. Individuals of the species were sexed and tagged during spawning season for subsequent monitoring to determine possible sex changes. Mortality was minimized by following a carefully designed sexing protocol. Out of 49 tagged specimens (41–88mm in size) that could be sexed in successive years (28 males and 21 females), 16 males changed into females between consecutive years (50.0% of males smaller than 70mm and 100% of males larger than this size), while two females changed sex to males between consecutive spawning seasons (both were smaller than 70 mm). Overall, 36.7% of the limpets monitored changed sex between consecutive years: 57.1% of males became females and 9.5% of females changed to males These observations confirms the occurrence of two-way sex change, or reverse sequential hermaphroditism, in P. ferruginea. Our findings unveil this sexual strategy in this endangered limpet and provide new direction for studies designed to address the mechanisms and factors that determine sex change and its effects on population dynamics., This work was funded at its late stage by the project “Action plan for viability proposals of the endangered limpet, Patella ferruginea” (Project 0 of the Spanish Research Council –CSIC- Foundation) and in former stages by grants awarded by the Organismo Autónomo de Parques Nacionales and the Dirección General de Conservación de la Naturaleza of the Spanish Ministerio de Medio Ambiente.

We modelled the distribution of two vulnerable steppe birds, Otis tarda and Tetrax tetrax, in the Western Palearctic and projected their suitability up to the year 2080. We performed two types of models for each species: one that included environmental and geographic variables (space-included model) and a second one that only included environmental variables (space-excluded model). Our assumption was that ignoring geographic variables in the modelling procedure may result in inaccurate forecasting of species distributions. On the other hand, the inclusion of geographic variables may generate an artificial constraint on future projections. Our results show that space-included models performed better than space-excluded models. While distribution of suitable areas for T. tetrax in the future was approximately the same as at present in the space-included model, the space-excluded model predicted a pronounced geographic change of suitable areas for this species. In the case of O. tarda, the space-included model showed that many areas of current presence shifted to low or medium suitability in the future, whereas a northward expansion of intermediate suitable areas was predicted by the space-excluded one. According to the best models, current distribution of these species can restrict future distribution, probably due to dispersal constraints and site fidelity. Species ranges would be expected to shift gradually over the studied time period and, therefore, we consider it unlikely that most of the current distribution of these species in southern Europe will disappear in less than one hundred years. Therefore, populations currently occupying suitable areas should be a priority for conservation policies. Our results also show that climate-only models may have low explanatory power, and could benefit from adjustments using information on other environmental variables and biological traits; if the latter are not available, including the geographic predictor may improve the reliability of predicted results., AE has a contract funded by the project 1098/2014 (Organismo Autónomo Parques Nacionales, Spain). This paper is a contribution to CGL2009-13029/BOS of the Spanish Ministry of Science, as well as to the REMEDINAL 3 (S2013/MAE-2719) network of the CAM. This work was partly supported by the Project CGL2009/11316/BOS (Spanish Ministry of Science and FEDER).

Seabirds are colonial vertebrates that despite their great potential for long-range dispersal and colonization are reluctant to establish in novel locations, often recruiting close to their natal colony. The foundation of colonies is therefore a rare event in most seabird species and little is known about the colonization process in this group. The Cory's shearwater (Calonectris diomedea) is a pelagic seabird that has recently established three new colonies in Galicia (NE Atlantic) thus expanding its distribution range 500 km northwards. This study aimed to describe the establishment and early progress of the new Galician populations and to determine the genetic and morphometric characteristics of the individuals participating in these foundation events. Using 10 microsatellite loci, we tested the predictions supported by different seabird colonization models. Possibly three groups of non-breeders, adding up to around 200 birds, started visiting the Galician colonies in the mid 2000's and some of them eventually laid eggs and reproduced, thus establishing new breeding colonies. The Galician populations showed a high genetic diversity and a frequency of private alleles similar to or even higher than some of the large historical populations. Most individuals were assigned to several Atlantic populations and a few (if any) to Mediterranean colonies. Our study suggests that a large and admixed population is settling in Galicia, in agreement with predictions from island metapopulation models of colonization. Multiple source colonies imply that some birds colonizing Galicia were dispersing from very distant colonies (> 1500 km). Long-distance colonizations undertaken by relatively large and admixed groups of colonizers can help to explain the low levels of genetic structure over vast areas that are characteristic of most oceanic seabird species., This study was funded annually (2010–2013) by the Organismo Autónomo de Parques Nacionales of Spain (http://www.magrama.gob.es/es/parques-nacionales-oapn/programa-investigacion/) through the research project 079/2009. Financial support for the fieldwork in Azores, Madeira, Selvagens and Berlengas was provided by the EU project LIFE04NAT/PT/000213, coordinated by SPEA (Sociedade Portuguesa para o Estudo das Aves), the BirdLife International partner in Portugal. HP acknowledges the support given by the Fundação para a Ciência e Tecnologia (Portugal, SFRH/BD/21557/2005 and SFRH/BPD/85024/2012). Financial support for the fieldwork in Cima Islet was provided by the EU project LIFE09NAT/PT/000041

The study of the factors structuring genetic variation can help to infer the neutral and adaptive processes shaping the demographic and evolutionary trajectories of natural populations. Here, we analyse the role of isolation by distance (IBD), isolation by resistance (IBR, defined by landscape composition) and isolation by environment (IBE, estimated as habitat and elevation dissimilarity) in structuring genetic variation in 25 blue tit (Cyanistes caeruleus) populations. We typed 1385 individuals at 26 microsatellite loci classified into two groups by considering whether they are located into genomic regions that are actively (TL; 12 loci) or not (NTL; 14 loci) transcribed to RNA. Population genetic differentiation was mostly detected using the panel of NTL. Landscape genetic analyses showed a pattern of IBD for all loci and the panel of NTL, but genetic differentiation estimated at TL was only explained by IBR models considering high resistance for natural vegetation and low resistance for agricultural lands. Finally, the absence for IBE suggests a lack of divergent selection pressures associated with differences in habitat and elevation. Overall, our study shows that markers located in different genomic regions can yield contrasting inferences on landscape-level patterns of realized gene flow in natural populations., This study was funded by grants 69/2003(Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia), CGL2010-21933-C02-01(Ministerio de Ciencia e Innovación) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund). ESF and JBE are supported by a PhD scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund, VGN is supported by a Forschungskredit of the University of Zurich (FK-14-103), RB is supported by a Juan de la Cierva fellowship (Ministerio de Ciencia e Innovación), ESD is supported by a PhD scholarship from Ministerio de Ciencia e Innovación-European Social Fund (FPIProgram), and JO is supported by research contracts funded by Severo Ochoa (SEV-2012-0262) and Ramón y Cajal (RYC-2013-12501) Programs.

Daily activity patterns in mammals depend on food availability, reproductive stage, habitat selection, intraspecific interactions and predation risk, among other factors. Some mammals exhibit behavioral plasticity in activity patterns, which allows them to adapt to environmental changes. A good example of this can be found in the red fox Vulpes vulpes. This species is adapted to living in highly humanized environments, where it is often culled because it may affect human interests (e.g. through the consumption of game species or livestock). We assessed the potential main drivers of the daily activity patterns of the red fox in 12 Iberian Mediterranean areas through the use of camera traps. Among drivers, we considered main prey availability (European wild rabbits Oryctolagus cuniculus), degree of human disturbance (e.g. distance to human settlements, and intensity of predator control) and habitat structure. Our results revealed a predominantly crepuscular and nocturnal activity of foxes with local variations. Although overall fox activity increased with rabbit availability, the temporal overlap with prey activity was on average low, because foxes increased activity when rabbits decreased theirs (twilight-night). Red fox activity rhythms seemed to be determined by human presence where human disturbance is high. In addition, diurnal activity decreased in areas with higher levels of human disturbance (closer to human settlements and high predator control intensity) and increased in dense habitats. Our study shows that daily activity patterns of highly adaptable species are determined by several interacting drivers, resulting in complex behavioral patterns. This suggests that further ecological studies should consider different factors simultaneously for a better understanding of daily activity patterns of wildlife in different landscapes., This study was funded by projectsCGL2009-10741 (Spanish Ministry of Science and Innovation and EU-FEDER funds); HUNTing for Sustainability(212160, FP7-ENV-2007-1), and OAPN352/2011 (Organismo Autónomo de Parques Nacionales, Spain). F. Díaz-Ruiz and J. Caro enjoyed postdoctoral contracts financedby the European Social Fund (ESF) and the Junta de Comunidades de Castilla-La Mancha (Operational Programme FSE 2007/2013). M. Delibes-Mateos is currently funded by Consejería de Economía, Innovación, Ciencia y Empleo of Junta de Andalucía, and the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement 267226.

We test the use of hyperspectral sensors for the early detection of the invasive dense-flowered cordgrass (Spartina densiflora Brongn.) in the Guadalquivir River marshes, Southwestern Spain. We flew in tandem a CASI-1500 (368–1052 nm) and an AHS (430–13,000 nm) airborne sensors in an area with presence of S. densiflora. We simplified the processing of hyperspectral data (no atmospheric correction and no data-reduction techniques) to test if these treatments were necessary for accurate S. densiflora detection in the area. We tested several statistical signal detection algorithms implemented in ENVI software as spectral target detection techniques (matched filtering, constrained energy minimization, orthogonal subspace projection, target-constrained interference minimized filter, and adaptive coherence estimator) and compared them to the well-known spectral angle mapper, using spectra extracted from ground-truth locations in the images. The target S. densiflora was easy to detect in the marshes by all algorithms in images of both sensors. The best methods (adaptive coherence estimator and target-constrained interference minimized filter) on the best sensor (AHS) produced 100% discrimination (Kappa = 1, AUC = 1) at the study site and only some decline in performance when extrapolated to a new nearby area. AHS outperformed CASI in spite of having a coarser spatial resolution (4-m vs. 1-m) and lower spectral resolution in the visible and near-infrared range, but had a better signal to noise ratio. The larger spectral range of AHS in the short-wave and thermal infrared was of no particular advantage. Our conclusions are that it is possible to use hyperspectral sensors to map the early spread S. densiflora in the Guadalquivir River marshes. AHS is the most suitable airborne hyperspectral sensor for this task and the signal processing techniques target-constrained interference minimized filter (TCIMF) and adaptive coherence estimator (ACE) are the best performing target detection techniques that can be employed operationally with a simplified processing of hyperspectral images., This study has been funded by the Spanish Ministry of Science and Innovation through the research projects HYDRA (No. CGL2006-02247/BOS) and HYDRA2 (CGL2009-09801/BOS), by the National Parks Authority (Organismo Autonomo de Parques Nacionales) of the Spanish Ministry of Environment to project OAPN 042/2007, and by funding from the European Union (EU) Horizon 2020 research and innovation program under grant agreement No. 641762 to ECOPOTENTIAL project. The Espacio Natural de Doñana provided permits for field work in protected areas with restricted access. We are grateful to the Instituto Nacional de Técnica Aeroespacial (INTA), Spain, for performing the airborne campaign and the geometric correction of the images. J.B. has to acknowledge a sabbatical stay at Pye Laboratory of the Commonwealth Scientific and Research Organization (CSIRO) Marine and Atmospheric Sciences, Australia, and at the Climate Change Cluster (C3) of the University of Technology Sydney, Australia, funded by the Spanish Ministry of Education, during data analysis and writing of this paper. This publication is a contribution from CEIMAR and also a contribution from CEICAMBIO. We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)

[EN]: Iberian wild goat subsp. Capra pyrenaica victoriae inhabits fewer and smaller areas than subs. C. p. hispanica reason why the former is considered as Vulnerable by IUCN. Therefore studies aimed to assess the status of C. p. victoriae are very valuables. We studied the habitat use, parasite excretion rates and diet quality of the Montes de Toledo's C. p. victoriae population. The red deer is a potential competitor of the Iberian wild goat and subsequently was included in this study. Our results showed that both species had a good population status (good corporal condition, high quality diet and suitable population density). According to the indicators considered in this study we do not expect that red deer suppose a relevant risk for the viability of the Iberian wild goat population. The implications of the results to Iberian wild goat management are deeper discussed., [ES]: La Capra pyrenaica victoriae presenta pocas poblaciones y éstas ocupan áreas reducidas, siendo considerada por ello por la IUCN como vulnerable. Son necesarios, por tanto, estudios locales encaminados a evaluar el estado de dichas poblaciones. En este trabajo estudiamos la población presente en los Montes de Toledo a través del análisis del uso que hace del hábitat, del nivel de parasitación y de la calidad de su dieta. El ciervo fue también incluido en este esquema de monitorización por ser un potencial competidor para la cabra montés. Los resultados indican que ambas especies presentan un adecuado estado poblacional (buena condición corporal, dieta de elevada calidad y valores de densidad razonables). Según los indicadores considerados no sería esperable que el ciervo supusiera un riesgo relevante para la viabilidad de la población de cabra montés. Se discuten las implicaciones de los resultados para la gestión cinegética de la cabra montés., Este trabajo ha sido promovido y financiado por el Parque Nacional de Cabañeros (Organismo Autónomo de Parques Nacionales) en el marco del convenio de colaboración 8.06/5.46.3324. P. Acevedo disfruta de un contrato dentro del programa Juan de la Cierva – Fondo Social Europeo.

The study of the genetic consequences of dispersal is a central topic in evolutionary, conservation and behavioural research. However, few studies have simultaneously considered dispersal movements from marked individuals and contemporary patterns of gene flow. We analysed the link between dispersal behaviour and gene flow in four populations of blue tits with different degrees of connectivity. For this purpose, we monitored four breeding patches and used genotypic and capture–mark–recapture data to study the genetic consequences of dispersal at different spatial scales. Data on natal dispersal movements revealed that both males and females dispersed less than expected under a random pattern of settlement at the two large spatial scales considered: the whole study area and the two main localities. However, natal dispersal distance was lower than expected under random settlement within natal patches in males whereas an opposite pattern was found for females. Accordingly, microsatellite data revealed limited gene flow between the localities studied and an isolation-by-distance pattern of genetic structure that was particularly strong at the large spatial scale (i.e. considering geographically distant breeding patches). Finally, the strong male philopatry was reflected by a stronger genetic structure and a lower admixed ancestry in this sex. Overall, we found evidence that restricted dispersal and fragmentation may have both contributed to reduce interpopulation gene flow at different spatial scales in a forest species and that there is concordance between genetic studies and those based on capture–mark–recapture., During this work V.G.N. and J.O. were supported by respective predoctoral FPI (MEC) and postdoctoral JAE-Doc (CSIC) fellowships. This study was funded by the grants 69/2003 (Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund).

This study aimed to maximize the sensitivity of bovine tuberculosis detection in living wild fallow deer (Dama dama) under field conditions. We evaluated the rapid test (RT; CervidTB STAT-PAK Assay, Chembio Diagnostic Systems, Inc., USA) in comparison with the comparative cervical skin test (CCT). A total of 134 fallow deer were captured between January and March 2008. At time 0, 0.1 ml of avian purified protein derivative (avian PPD; Cooper-Zeltia, Spain), 0.1 ml bovine PPD (Cooper-Zeltia, Spain), 0.1 ml negative control PBS and 0.1 ml of a positive control (the mitogen phytohaemagglutinin, PHA; containing 250 mg PHA, diluted in PBS) were injected intradermally at four shaved sites in the neck. The skin fold thickness at each injection site was measured at time 0 and 72 h (3 repeats each time). Animals with a skin test response of 2 mm or more at the bovine PPD injection site and animals with any visible reactivity in the RT were necropsied and tissues submitted for culture and for histopathology. A total of 36 fallow deer were considered reactors to bovine PPD or to the RT (apparent prevalence 27%). Regarding both bovine PPD reactivity and the skin fold increase at the PHA injection site, we found significant effects of age and sex by age interaction. Adult males had the largest responses. Mycobacterium bovis was isolated from lymphoid tissues of 21 fallow deer. Skin test sensitivity, as compared to M. bovis culture confirmed deer, was 80.1% (17/21). But, the CCT alone would have missed 4 of 21 culture confirmed animals. RT sensitivity, based on culture confirmed deer, was also 80.1% (17/21). Similarly, the RT alone would have missed another 4 of 21 culture confirmed deer. However, combining the CCT and the RT allowed for detecting all 21 culture positive fallow deer. We conclude that the combined application of the RT and the skin testing can maximize the sensitivity of bTB detection in living fallow deer, thus facilitating control programs for wildlife disease surveillance., This study was funded by Patrimonio Nacional (special thanks to M. Moreno and A. Jacoste) through contracts to IREC and TRAGSEGA (A. Rosón). This is a contribution to MCINN Plan Nacional Research grant AGL2008-03875 and FEDER, and to EU grant TB-STEP 212414. Studies on diseases shared between domestic animals and wildlife are also supported by grants and contracts from INIA and Ministerio de Medio Ambiente y Medio Rural y Marino (OAPN and SDGSPP), and Grupo Santander – Fundacion Marcelino Botin. MPM-H acknowledges a post-doctoral contract from Instituto de Salud Carlos III.

9 páginas, 5 figuras, 4 tablas.-- Este artículo está dedicado a nuestro amigo y colega el Prof. Alejandro Javalera, fallecido en el curso de esta investigación., This study shows results on litterfall dynamics and decay in mangrove stands of Avicennia germinans distributed along a latitudinal gradient (three forest sites) in the Gulf of California, in order to assess whether internal sources could support the observed mangrove ecosystem organic deficit in this arid tropic. Total mean annual litterfall production increased southward (712.6 ± 53.3, 1501.3 ± 145.1 and 1506.2 ± 280.5 g DW m−2 y−1, in the Yaqui, Mayo and Fuerte areas respectively), leaves being the main component of litter in all locations during the entire year, followed by fruits. The wet season (June–September) showed the highest litterfall rates through fruits. The temporal trend of litterfall production was significantly explained through mean air temperature (R2 = 68%) whilst total annual litter production in the entire region showed a statistically significant relationship with total soil phosphorus, salinity, total nitrogen, organic matter and tree height (R2 = 0.67). Throughout 117 days of the decomposition experiment, the litter lost 50% of its original dry weight in 5.8 days (average decay rate of 0.032 ± 0.04 g DW d−1) and there were not significant differences in the remaining mass after 6 days. The percentage of both C and P released from the litter correlated significantly with the ratio of tidal inundated days to total experiment days (R2 = 0.62, p = 0.03 and R2 = 0.67, p = 0.02, respectively); however, the frequency of tidal inundation only showed a significant increase in C release from Avicennia litter after 6 and above 48 days of decomposition. Whereas the total C content of litter bags decreased linearly over the decomposition to (% Total C = 5.52 − 0.46 days, R2 = 0.81, p = 0.0005), N content displayed an irregular pattern with a significant increase of decay between 48 and 76 days from the beginning of the experiment. The pattern for relative P content of litter revealed reductions of up to 99% of the original (%tot-P = −9.77 to 1.004 days, R2 = 0.72, p = 0.01) although most of the P reduction occurred between 17 and 34 days after the experiment started. Soil N and P contents, which exhibited significant differences in the course of the decomposition experiment, appeared to show significant differences between sampling sites, although they were not related to tidal influence, nor by leaf and nutrient leaching. In a global basis, C/N litter ratios decreased linearly (C/N = 32.86 − 0.1006 days, R2 = 0.62, p = 0.02), showing a strong and significant correlation with meteorological variables (R2 = 0.99, p = 0.01). C/P ratios of litter increased through an exponential function (C/P = 119.35e0.04 day, R2 = 0.89, p < 0.001). Changes in the remaining percentage of litter mass during the experiment were significantly correlated with soil C/N ratio (R2 = 0.56, p = 0.03) as well as with the soil C/P ratio (R2 = 0.98, p < 0.001). Our results of litter decomposition dynamics in this mangrove support the fact of null net primary productivity of the arid mangrove wetlands: fast litter decomposition compensates the ecosystem organic deficit in order to sustain the mangrove productivity. Litter decomposition plays a key role in the ecosystem metabolism in mangroves of arid tropics., This research was supported by the Mexican Secretary of Natural Resources and Environment (SEMARNAT-CONACYT; contract SEMARNAT-2002-C01-0147) and by the Spanish Ministry of Environment (Organismo Autónomo Parques Nacionales; contract 2008/001 DECAMERON). The first author has been granted a CSIC-JAEDoc contract.

Serum samples from 101 mouflons (Ovis aries musimon) collected from July 2002 to January 2006 were tested for antibodies against Anaplasma spp., Brucella spp., bovine viral diarrhea virus, Chlamydophila abortus, Coxiella burnetii, Mycobacterium avium ssp. paratuberculosis, and Maedi-Visna virus. Mouflon came either from extensive farms or high ungulate density fenced hunting estates. Antibodies were detected against Anaplasma spp. (22.2%), C. burnetii (4.0%), M. avium ssp. paratuberculosis (1.0%) and C. abortus (1.0%). According to our results, mouflons could participate as a wild reservoir in the epidemiology of Anaplasma spp. infection and maybe Q fever, but they do not seem to contribute in the epidemiology of the rest of the studied infectious diseases in south-central Spain., This is a contribution to the agreement CSIC–SDGSA–OAPN and to agreements with Consejería de Agricultura, JCCM. J. R. López-Olvera has an I3P grant from the Consejo Superior de Investigaciones Científicas (CSIC).

Viral diseases can influence the population dynamics of wild carnivores and can have effects on carnivore conservation. Hence, a serologic survey was conducted in an opportunistic sample of 137 foxes (Vulpes vulpes) and 37 wolves (Canis lupus) in Spain for 1997–2007 to detect antibodies against canine distemper virus (CDV) and against canine parvovirus (CPV) by indirect ELISA. Antibodies against CDV were detected in 18.7% of the analyzed animals and antibodies against CPV in 17.2%. There was no difference in antibody prevalence to CDV between both species, even in the same region (P > 0.05), but there was a significant difference in antibody prevalence to CPV between foxes (5.1%) and wolves (62.2%) (P < 0.05). In fox populations there was a significant difference in antibody prevalence to CDV between geographic areas (Aragón 26.4%, La Mancha 7.8%, P < 0.05). In wolf populations there was significantly higher antibody prevalence against CPV (P < 0.05) in Castilla y León (100%) than in the Cantabric region (53.3%). There was no significant sex or age-related difference in the antibody prevalence against CDV or CPV in foxes. These results indicate that contact with CDV is widespread among wild canid populations in Spain and that CPV is endemic in the Iberian wolf population. The implications of these results are briefly discussed., This is a contribution to the agreement between IREC, SDGSA-MAPA and OAPN-MMA, and to the agreement between CSIC and Principado de Asturias. RS Acknowledges a grant from Castilla-La Mancha.

Serum samples from 282 wild carnivores from different regions of Spain were tested for antibodies to Toxoplasma gondii by the modified agglutination test using a cut-off value of 1:25. Antibodies to T. gondii were found in 22 of 27 (81.5%) of Iberian lynx (Lynx pardinus), 3 of 6 European wildcats (Felis silvestris), 66 of 102 (64.7%) red foxes (Vulpes vulpes), 15 of 32 (46.9%) wolves (Canis lupus), 26 of 37 (70.3%) Eurasian badgers (Meles meles), 17 of 20 (85.0%) stone martens (Martes foina), 4 of 4 pine martens (Martes martes), 6 of 6 Eurasian otters (Lutra lutra), 4 of 4 polecats (Mustela putorius), 1 of 1 ferret (Mustela putorius furo), 13 of 21 (61.9%) European genets (Genetta genetta), and 13 of 22 (59.1%) Egyptian mongooses (Herpestes ichneumon). Serological results indicated a widespread exposure to T. gondii among wild carnivores in Spain. The high T. gondii seroprevalence in Iberian lynx and the European wildcat reported here may be of epidemiologic significance because seropositive cats might have shed oocysts., This study is a contribution to GC-05-006, ICS, JCCM, and agreements between IREC and Principado de Asturias and Ministerio de Agricultura and OAPN, Spain. This study received partially support from the Spanish CICYT, grants AGL2004-06103-C02-01/GAN and AGL2005-07401. This study was also partially included in the “Programa de Actuaciones para la Conservación del Lince en Andalucía II”. Consejería del Medio Ambiente, Junta de Andalucía. R Sobrino was recipient of a PhD grant from Junta de Comunidades de Castilla La Mancha (JCCM).

There has been a worldwide increase in the number and geographical spread of wild boar populations in recent decades leading to an increase in both the circulation of disease agents and greater contact with domestic animals and humans. Diseases affect the population dynamics of wildlife but the effects of most viral diseases on the European wild boar are largely unknown. Many viral diseases present in domestic pig populations are also present in wild boars where they can provide a disease reservoir, as is clearly the case with classical swine fever, but little is known about other viral diseases such as porcine circovirus diseases or hepatitis E. This review considers the current scientific knowledge of the effects of viral diseases on wild boar populations and their rôle as potential disease reservoirs. The focus is on those viral diseases of domestic swine and wild boars that are included as notifiable by the Office International des Epizooties (OIE)., This is a contribution to the agreement between MAPA (SDGSA), OAPN and IREC.

The embryonic development of the squid Loligo vulgaris was observed from 183 egg masses collected from special devices deployed throughout Cabrera National Park (Baleares Islands, western Mediterranean Sea). Sequence alignment analysis of the cytochrome oxidase I gene revealed that all embryos belonged to L. vulgaris. In total, 549 egg capsules were examined. Viable egg capsules (n = 420) were classified into one of five maturation stages according to the primary external features. The length of the viable egg capsules varied between 40 and 170 mm, and increased with embryonic development. The non-viable capsules (n = 129) were categorized into four groups: I (Ginger root), non-viable II and III, and empty egg capsule (IV). The percentage of non-viable capsules (i.e. grades I, II and III) was 92.25%. Empty capsules accounted for 7.75% of the total non-viable egg capsules. Embryonic development was classified into a second scale of eight stages. Egg capsule stage and embryonic stage were significantly related (n = 420; p, This work was financially supported through the Organismo Autónomo de Parques Naturales de España of the Ministerio de Agricultura, Alimentación y Medio Ambiente (CEFAPARQUES, Project number: 458/2011)

Despite the importance of spring freezing events for alpine species distribution, few studies have analysed the response of alpine shrub species to early spring freezes. It is also not known how snow cover gradients influence the process of de-hardening between individuals of the same species and their vulnerability to early spring frosts. We analysed early spring freezing resistance for the buds of eight alpine Ericaceae shrubs growing at 1 m snow depth at treeline in the Swiss Alps. Moreover, buds of Rhododendron ferrugineum and Loiseleuria procumbens were analysed for freezing resistance and sugar content along a snow depth gradient. The LT50 (lethal temperature for 50% of samples) of the eight species ranged from −25.1 ± 1.6 °C (Vaccinium vitis-idaea) to −11.1 ± 1.2 °C (Vaccinium uliginosum), with differences being related to the phenological stage in addition to shrub preferences for contrasting snow cover microsites. Although the effect of snow depth on the freezing resistance of plants was not significant, samples collected from 1 m to 1.5 m snow depth tended to be more vulnerable to freezing, particularly L. procumbens. Buds collected at shallower snow depths had higher sugar concentrations, indicative of their stronger physiological hardening. Consequently, we conclude that differences in snow cover may significantly affect the physiological hardening of plants during the onset of spring. Individuals growing at less than 0.5 m snow cover are hardier, i.e., show moderately higher freezing resistance than individuals from snow banks. Snow cover is a highly important aspect of climate change, and freezing resistance in alpine plants with respect to snow conditions can be a relevant driver of plant responses to climate change., The authors thank Melisa Dawes, Christopher Hughes and Elisabeth Leibold, for assistance during field work, Britta Jahn-Humphrey for help in the lab, Julia Wheeler for assistance with plant processing and English editing and Kari Taulavuori and two anonymous referees for useful comments on earlier versions of the manuscript. SP was funded by the Ramón y Cajal fellowship (RYC-2013-14164, Ministerio de Economía y Competitividad, Spain), Arbaltmont project (Organismo Autónomo de Parques Nacionales, Spain) and DGA (Gobierno de Aragón) travel grant (BOA 9/4/2012).

We carried out field studies and laboratory experiments to investigate (1) the possible feeding preference of the red swamp crayfish Procambarus clarkii on living macrophytes, and (2) the influence that such a feeding preference can cause on the distribution and bundance of P. clarkii in wetland systems. Field studies, at two sampling areas of the Spanish wetland Tablas de Daimiel National Park (TDNP), showed that P. clarkii had significantly higher mean values of density and biomass at S-2 (with Char a hispida as the dominant macrophyte) than at S-1 (with Ceratophyllum submersum as the dominant macrophyte). Laboratory experiments showed that P. clarkii had a significant feeding preference for C. hispida versus C. submersum. Analyses of the biochemical composition of each macrophyte species showed that the unpreferred macrophyte (C. submersum) had higher amounts (per unit of biomass) of total protein, nitrogen, phosphorus, sodium, potassium, magnesium and phenolic compounds than the preferred macrophyte (C. hispida). By contrast, C. hispida had more calcium per unit of biomass than C. submersum. Overall, we conclude that the presence of higher amounts of phenolic compounds in C. submersum might be the foremost factor responsible for the observed feeding preference of P. clarkii on living macrophytes in TDNP., This research was funded by the Autonomous Organism for National Parks, Ministry of the Environment, in Spain.

[Aim] Conifers comprise an ancient and diverse group of plants showing a wide distribution range. To better understand the general patterns of species successfully established on islands, this review compiles information about the distribution, diversity, dispersal potential and conservation status of insular conifers, with special emphasis on those inhabiting remote oceanic islands., [Methods] An exhaustive survey was made of world‐wide databases and literature. We registered information on island distribution (including ocean region, extension and geological origin of the island), endemism and threat status for each insular conifer., [Main conclusion] Our results highlight the wide diversity of insular conifers, as well as the key role of oceanic islands in catalysing speciation mechanisms. Pacific islands in particular harbour the greatest diversity levels, constituting a major centre of diversification. The wide distribution of conifers reflects their great potential for dispersal and colonization, endozoochory being the most favourable dispersal syndrome for reaching remote islands. The general threat status of insular conifers highlights the fragility of island biota and the urgent need for policies focused on their preservation., [Results] 285 of the 547 conifer species considered in this review show an insular distribution (i.e. their distribution encompass insular territories). The family Podocarpaceae is best represented, with 40% of the insular species. The importance of endozoochory for long‐distance dispersal is clear, because it was the most frequent dispersal syndrome among oceanic conifers. A high proportion of the total threatened conifers occur on islands (52%), and many of them are insular endemics (72%). Among conifer families, Araucariaceae is the most threatened in insular territories., This work has been supported by the Ministerio de Educación y Ciencia (CGL 2010‐18759) and the Organismo Autónomo de Parques Nacionales (project 051/2010).