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Environmental DNA (eDNA) monitoring has revolutionized the way biodiversity is surveyed and has been proposed as a reliable method to inform management decisions. Recognition of eDNA techniques as reliable tools to inform management and biosecurity require stringent standards to assess sample quality and reliability of results. Laboratories can have their workflows assessed and certified through accreditation and be involved in proficiency testing schemes provided to test the accuracy and precision of molecular methods. Currently, there is only one eDNA‐based proficiency testing scheme designed to test competency of laboratories in amplifying eDNA from the Great Crested Newt, Triturus cristatus (Laurenti, 1768) in water samples. This test, however, is a closed scheme currently run by invitation only to laboratories in the United Kingdom. Given the paucity of eDNA‐based proficiency schemes to ensure high‐quality services, this commentary discusses how future proficiency testing schemes could be designed to assess eDNA sample quality and reliability on detection results for environmental management and biosecurity applications. We discuss the use of tissue‐derived and synthetic oligonucleotides as reference materials, the need for proficiency testing schemes to assess the capacity of analytical facilities to determine sample quality as well as accurately detecting trace eDNA in blind samples and discuss the context in which fit for purpose eDNA testing schemes could be designed. To complement the future development of eDNA proficiency testing schemes, we provide firsthand accounts and lessons learned while developing the current Great Crested Newt eDNA proficiency scheme. Lastly, we highlight current limitations in standardizing rapidly improving eDNA‐based techniques and discuss disadvantages to accreditation and standardization from an Australian‐centered perspective as a means to promote an active debate on the topic of future eDNA accreditation and proficiency testing.

Terrestrial landscapes surrounding aquatic habitat influence the persistence of amphibian spatially structured populations (SSPs) via their crucial role in providing estivation and overwintering sites, facilitating or hampering dispersal and colonisation, and consequently the maintenance or loss of genetic diversity. To highlight the landscape drivers of genetic variation, we investigated the relationship between the level of genetic variation measured within ponds of the great crested newt (Triturus cristatus), and the composition of the surrounding landscape at various spatial scales. Based on the sampling of 40 ponds in 13 SSPs, the influence of landscape features on several estimators of genetic variation was investigated via linear mixed models, with effects within and between SSPs incorporated. The best models depended on the spatial scale, with more significant associations within radii of 50 and 100 m of core ponds, particularly for allelic richness. Responses within and between SSPs were mostly similar. The availability of aquatic habitat in the landscape had a positive effect, while woodland, arable land and pasture had different effects depending on scale and response variable. Total length of roads within a 250 m radius influenced effective population size negatively. Our results stress the need to investigate the influence of environmental predictors at multiple spatial scales for an adequate understanding of ongoing processes. Generally, the landscape affected genetic variation similarly within and between SSPs. This allowed us to provide general guidelines for the persistence of great crested newt populations, with an emphasis on the importance of the aquatic habitat.

Decades of environmental DNA (eDNA) method application, spanning a wide variety of taxa and habitats, has advanced our understanding of eDNA and underlined its value as a tool for conservation practitioners. The general consensus is that eDNA methods are more accurate and cost‐effective than traditional survey methods. However, they are formally approved for just a few species globally (e.g., Bighead Carp, Silver Carp, Great Crested Newt). We conducted a meta‐analysis of studies that directly compare eDNA with traditional surveys to evaluate the assertion that eDNA methods are consistently “better.”Environmental DNA publications for multiple species or single macro‐organism detection were identified using the Web of Science, by searching “eDNA” and “environmental DNA” across papers published between 1970 and 2020. The methods used, focal taxa, habitats surveyed, and quantitative and categorical results were collated and analyzed to determine whether and under what circumstances eDNA outperforms traditional surveys.Results show that eDNA methods are cheaper, more sensitive, and detect more species than traditional methods. This is, however, taxa‐dependent, with amphibians having the highest potential for detection by eDNA survey. Perhaps most strikingly, of the 535 papers reviewed just 49 quantified the probability of detection for both eDNA and traditional survey methods and studies were three times more likely to give qualitative statements of performance. Synthesis and applications: The results of this meta‐analysis demonstrate that where there is a direct comparison, eDNA surveys of macro‐organisms are more accurate and efficient than traditional surveys. This conclusion, however, is based on just a fraction of available eDNA papers as most do not offer this granularity. We recommend that conclusions are substantiated with comparable and quantitative data. Where a direct comparison has not been made, we caution against the use of qualitative statements about relative performance. This consistency and rigor will simplify how the eDNA research community tracks methods‐based advances and will also provide greater clarity for conservation practitioners. To this end suggest reporting standards for eDNA studies., A meta‐analysis shows that environmental DNA outperforms traditional methods for biodiversity monitoring, both in terms of cost and efficiency.

1.It is of vital importance that aquatic conservation is evidence based, and in the field of farmland pond management and restoration evidence was largely lacking until an article published in Aquatic Conservation: Marine and Freshwater Ecosystems (AQC) in 2012. 2.To examine the influence of farmland pond management on aquatic biodiversity conservation, macrophyte and invertebrate diversity in ponds subject to traditional management involving scrub and occasional sediment removal at different time intervals in the past (0–2, 3–5, 6–10 years since management), were compared with a set of neighbouring, highly terrestrialized ponds that had not been managed for many decades. 3.With the exception of Mollusca, significantly higher species diversity was found for managed ponds compared with the late‐succession unmanaged ponds, with invertebrate gamma diversity significantly lower for the late‐succession ponds, compared with all the managed pond categories. 4.The AQC article was a key component of the Natural England (UK Government's adviser on the natural environment in England) ‘Freshwater and Wetland Conservation Narrative’ and has helped with integrating pond management into recent great crested newt Triturus cristatus mitigation policy as well as bringing it to the fore in past and evolving agri‐environment policy. 5.The AQC article provided the evidence and in turn the confidence for the authors and a number of conservation partners to form a Norfolk Ponds Project. Since 2014, the Project has delivered or facilitated more than 100 pond restorations in Norfolk, eastern England, as well as educating conservation practitioners and farmers on the importance of farmland ponds via various events. 6.With good underpinning science it has been possible to achieve considerable impact in the field of farmland pond conservation. This study shows the importance of setting aside time and support for academic staff to translate applied research outputs into practical impact.

Environmental DNA (eDNA) metabarcoding is revolutionising biodiversity monitoring, but has unrealised potential for ecological hypothesis generation and testing. Here, we validate this potential in a large-scale analysis of vertebrate community data generated by eDNA metabarcoding of 532 UK ponds. We test biotic associations between the threatened great crested newt (Triturus cristatus) and other vertebrates as well as abiotic factors influencing T. cristatus detection at the pondscape. Furthermore, we test the status of T. cristatus as an umbrella species for pond conservation by assessing whether vertebrate species richness is greater in ponds with T. cristatus and higher T. cristatus Habitat Suitability Index (HSI) scores. T. cristatus detection was positively correlated with amphibian and waterfowl species richness. Specifically, T. cristatus was positively associated with smooth newt (Lissotriton vulgaris), common coot (Fulica atra), and common moorhen (Gallinula chloropus), but negatively associated with common toad (Bufo bufo). T. cristatus detection did not significantly decrease as fish species richness increased, but negative associations with common carp (Cyprinus carpio), three-spined stickleback (Gasterosteus aculeatus) and ninespine stickleback (Pungitius pungitius) were identified. T. cristatus detection was negatively correlated with mammal species richness, and T. cristatus was negatively associated with grey squirrel (Sciurus carolinensis). T. cristatus detection was negatively correlated with larger pond area, presence of inflow, and higher percentage of shading, but positively correlated with HSI score, supporting its application to T. cristatus survey. Vertebrate species richness was significantly higher in T. cristatus ponds and broadly increased as T. cristatus HSI scores increased. We reaffirm reported associations (e.g. T. cristatus preference for smaller ponds) but also provide novel insights, including a negative effect of pond inflow on T. cristatus. Our findings demonstrate the prospects of eDNA metabarcoding for ecological hypothesis generation and testing at landscape scale, and dramatic enhancement of freshwater conservation, management, monitoring and research.

The study was focused on determining the sites of amphibian and reptile occurrence in the Pieprzowe Mountains Nature Reserve (area 18.01 ha) and adjacent areas (total area 58.81 ha). The investigations also involved the biology of breeding in the selected species, as well as determining threats and protection measures. Situated in the eastern part of the Świętokrzyskie Voivodeship, the steppe-like Pieprzowe (Pepper) Mountains Nature Reserve was established in 1979. Its aim is to protect xerothermic assemblages inhabited by many species of rare plants including the largest European aggregation of roses growing wild. The area is located on a steep, sometimes rugged escarpment, which is part of the Vistula River erosional margin. The escarpment exposes Cambrian black pepper-like shales. The slope base is overgrown by a narrow belt of willow thickets. Above occur assemblages of xerothermic thickets with a few trees. The highest parts of the nature reserve are overgrown with xerothermic grasslands with the prevalence of feathergrass steppe (Festuco-Stipion class). The reserve is surrounded by riparian forests, waterlogged meadows and reservoirs of the Vistula former riverbed, the largest of which is an oxbow lake (5.16 ha). The following species were documented in 2016-2017: smooth newt Lissotriton vulgaris L., great crested newt Triturus cristatus Laur., European fire-bellied toad Bombina bombina L., common toad Bufo bufo L., European green toad Bufotes viridis Laur., European tree frog Hyla arborea L., edible frog Pelophylax esculentus L., pool frog Pelophylax lessonae Cam., marsh frog Pelophylax ridibundus Laur., moor frog Rana arvalis Nilss., common frog Rana temporaria L., sand lizard Lacerta agilis L., grass snake Natrix natrix L. and common European adder Vipera berus L. The studies were focused on biology of breeding and phenology in common toad and common frog. The main threats posed on herpetofauna include: human presence, littering, fire raising and changes of water balance. © IOŚ-PIB

The distributions and conservation status of the six native amphibian species in Scotland are reviewed. Issues of under‐recording and false recording are noted, and steps to remedy these problems are described. Threats to amphibians in Scotland include diseases (a minor impact so far), road deaths, habitat loss, and climate change (to which populations may be resilient). Exploitation, competition, and pollution do not appear to be significant threats to amphibians in Scotland. Legal protection is currently strongest for the great crested newts (Triturus cristatus) and natterjack toads (Epidalea calamita), but the natterjack toads have only a small localized Scottish population. The ponds at Gartcosh hold the largest great crested newt population in Scotland. Translocation to a specially created reserve appears to have been successful but a new road across the site is an additional hazard, for which monitoring is in progress. Amphibian habitat enhancement in Scotland includes pond‐creation programmes, which have successfully increased the diversity of plants and invertebrates as well as amphibians. The value of Sustainable Drainage Systems (SuDS) for amphibians and other wildlife is reviewed. The Eurasian beaver reintroduction programme may benefit amphibians in the longer term, but research is currently lacking. Road tunnels combined with fences are a vital mitigation for road‐traffic impacts on amphibians and have been monitored at one Scottish site. Amphibian ladders in road drains are another potentially helpful mitigation. The amphibians of Scotland need more long‐term monitoring. A national strategy is imminent and should stimulate the required research and action.

Dispersal is a central process in ecology and evolution with far reaching consequences for the dynamics and genetics of spatially structured populations (SSPs). Individuals can adjust their decisions to disperse according to local fitness prospects, resulting in context-dependent dispersal. By determining dispersal rate, distance and direction, these individual-level decisions further modulate the demography, relatedness and genetic structure of SSPs. Here, we examined how context-dependent dispersal influences the dynamics and genetics of a great crested newt (Triturus cristatus) SSP. We collected capture-recapture data of 5564 individuals and genetic data of 950 individuals across an SSP in northern Germany. We added genetic data from six sites outside this SSP to assess genetic structure and gene flow at a regional level. Dispersal rates within the SSP were high but dispersal distances were short. Dispersal was context-dependent: individuals preferentially immigrated into high-quality ponds where breeding probabilities were higher. The studied SSP behaved like a patchy population, where subpopulations at each pond were demographically interdependent. High context-dependent dispersal led to weak but significant spatial genetic structure and relatedness within the SSP. At the regional level, a strong hierarchical genetic structure with very few first-generation migrants as well as low effective dispersal rates suggest the presence of independent demographic units. Overall, our study highlights the importance of habitat quality for driving context-dependent dispersal and therefore demography and genetic structure in SSPs. Limited capacity for long-distance dispersal seems to increase genetic structure within a population and leads to demographic isolation in anthropogenic landscapes.

In vertebrates, the main tissue devoted to energy storage is the adipose tissue. In salamanders, energy reserves can also be stored in the adipose tissues of the tail. Therefore, we evaluated if energy storage in salamanders' tails is related to individual body condition, life cycle and environmental constraints. We calculated a scaled measure of tail width for 345 salamanders belonging to six Mediterranean taxa exhibiting wide phylogenetic, behavioural and ecological variation. We related this measure to the Scaled Mass Index (SMI), a body condition index which reliably predicts body fat. We found significant relationships between the SMI and scaled tail width in the terrestrial Spectacled salamander and Alpine salamanders, independently of sex. At the same time, we found that energy storage in the tail is maximum in Alpine Salamanders, which experience reduced activity periods and restricted access to resources. Conversely, we found a significant effect of sex in Imperial cave salamanders, where females store reserves in the tail to counterbalance resource investment in parental care, and in Corsican Brook Newts, where the reproductive function of males' tails may imply a greater tail width. Finally, in the biphasic Great Crested Newt, tail width was not related to SMI in both sexes.

Simple Summary This research describes a successful protocol for establishing viable cell lines from one of the amphibian species that are severely threatened with extinction. For the study, fresh or cryopreserved tissue samples were used, and various parameters affecting the optimal growth conditions were tested, such as use of different media, impact of media change and the influence of contamination. We concluded that: (i) Using fresh tissue was more successful in terms of establishing cell lines; (ii) No differences were found between the media used for fresh tissue, while one medium had a positive effect on growth when using cryopreserved tissue; (iii) Contamination caused the tissue cultured to be destroyed; and (iv) Real-time cell analysis visualized a significant impact on cell growth pattern when media change was combined with PBS washing. This research demonstrated the potential of and the challenges to establishing amphibian cell lines, which provide an opportunity for implementing modern reproductive technologies as a future part of conservation strategies for endangered and threatened amphibian species. Abstract This study describes a successful protocol for establishing cell lines from the threatened Triturus cristatus in terms of collection, preparing, establishing, cryopreserving, thawing and quality checking. Different parameters such as media, media change, fresh vs. cryopreserved tissue and seeding density were tested to optimize culture conditions for this species. With fresh tissue, no considerable differences in the use of two different media were found, but with cryopreserved tissue, a combination of ITS (insulin/transferrin/selenite) and 2-mercaptoethanol had a positive effect on growth. Real-time measurements on the cell lines were used, for the first time in amphibian cells, to investigate the effect of different treatments such as media change with or without washing. Media change had a positive impact on the cells, whereas the effect was negative when combined with washing. It is concluded that establishment of cell lines is possible from the great crested newt, especially when using fresh tissue, but much more challenging if the tissue has been cryopreserved. Real-time measurement during cell culture is a useful tool to visualize the sensitivity of amphibian cells during different culture treatments.

International audience; Linear transport infrastructure can impact wildlife directly or indirectly, and amphibians are particularly affected. In some cases, mitigation measures e such as tunnels below the linear transport infrastructure e are implemented, but an evaluation of their effectiveness is often lacking, especially for small animals such as amphibians. Few studies have focused on crossing behaviour in underpasses, yet this information may be crucial to improve mitigation measures. In this study, we conducted experiments in three tunnels below a high-speed railway with individually marked amphibians (great crested newt Triturus cristatus, marbled newt Triturus marmoratus, alpine newt Ichthyosaura alpestris, fire sala- mander Salamandra salamandra, spined toad Bufo spinosus, agile frog Rana dalmatina and edible/pool frog Pelophylax kl. esculentus/lessonae) which were released at the entrance of tunnels varying in length. Behaviours (e.g. completed crossing, U-turn, not moving, and stay) and crossing speed were recorded. R. dalmatina, S. salamandra and T. cristatus (but not B. spinosus) exhibited crossing rate differences between tunnels 21 m and 40 m in length. More individuals performed “complete crossing” through the shorter tunnel. The crossing speed of the S. salamandra and T. cristatus did not differ between the tunnels of different lengths. In a second experiment, we evaluated the effect of acoustic enrichment on the crossing behaviour of one frog and three newt species by broadcasting a soundtrack of mating calls of syntopic anuran species in the tunnels. Pelophylax kl exhibited a large in- crease in complete crossings and in speed. T. cristatus showed an increase in tunnel complete crossings (but not speed) in one of the acoustically enhanced tunnels. These results indicate that acoustic enrichment can improve the crossing of underpasses. This low cost and easily implementable method could help to inform the design of wildlife passages as mitigation measures for impacted amphibian populations, though more ex- periments on the effect of tunnel characteristics on amphibian behaviour would be valuable.

Road mitigation infrastructure for pond‐breeding amphibians aims to provide a safe and sustainable passage for individuals between critical habitat patches. However, relatively little is known about how amphibians interact with mitigation systems because of the challenges of documenting movements at sufficiently large sample sizes. The effect of real or perceived barriers to short‐term movement could ultimately determine the success or failure of road mitigation schemes. We quantified behavioural responses of the protected great crested newt Triturus cristatus in a complex road mitigation system in the UK. We used fluorescent paint to mark individuals in order to measure distance travelled and trajectory orientation over two seasons (spring when adults migrate to breeding ponds and autumn when newts disperse) and in three components of the mitigation system (fences, tunnel entrances and inside the tunnels). A total of 250 juveniles and 137 adult great crested newts were marked and tracked during 38 survey nights. Adults were individually identified using belly‐pattern recognition. There was substantially greater activity along the fences during autumn (82% of newt captures) compared to spring. Triturus cristatus typically moved short distances each night (3.21 m per night in spring and 6.72 m per night in autumn), with a maximum of 25.6 m travelled inside a tunnel. Adult recapture rates were low (9.7%) and only 3% of the newts found along the fences reached the tunnel entrances. Movements were straighter in spring and inside the tunnels and newts had higher crossing rates in autumn compared to spring. Overall, behaviour and seasonal movement patterns significantly influenced the use of the mitigation system, in a way that could impact landscape connectivity for T. cristatus over the long‐term. Adequate incorporation of fine‐scale movement dynamics could help develop new behavioural models, inform our understanding of amphibian ecology and substantially improve future road mitigation projects.

The ecological niche of a species determines whether a species can persist and reproduce in a patch or not. The niche of a species is often described using habitat suitability models and indices. Accordingly, one may expect tight links between demography, phenotypes of individuals, population size, and habitat suitability. However, such links are not always found. Here, we study the relationship between a habitat suitability index that is commonly used for conservation assessments and metrics describing the performance at the level of populations and individuals. Using data from a metapopulation of a pond-breeding amphibian, the Great Crested Newt (Triturus cristatus), we show that habitat suitability predicts population size but not body condition. Ponds with higher suitability had a higher population size of newts, whereas population size correlated negatively with body condition of individuals. Our results are in line with previous studies showing no straightforward relationship between habitat suitability and body condition (a measure of individual performance) and the performance of populations. We suggest that a population size-dependent reduction of body condition may be a regulatory mechanism in newt populations.

Conserving genetic diversity in wild species is vital for preserving adaptations to local environmental conditions. We conducted a habitat creation project for a flagship European Protected Species of amphibian (northern great crested newt Triturus cristatus) at its north-western range edge in the Scottish Highlands, combining existing knowledge about the species’ local habitat requirements and the genetic structure of existing populations. Reconciled by a spatial connectivity model, we determined the best locations for the creation or restoration of 25 ponds to facilitate their natural colonisation, and to reconnect areas with similar genetic makeup whilst preserving differences between populations belonging to separate genetic clusters. A dynamic occupancy model based on data derived from standardised surveys revealed that both detection and occupancy probabilities strongly depended on habitat features known to be locally favourable for T. cristatus. At least 12 of the 25 new ponds were colonised within six years, representing a 26% increase in number of breeding ponds in the region and offsetting a gradual decline in number of pre-existing breeding sites recorded over the last 3 decades. Combining genetic evidence, spatial modelling and historical knowledge with local stakeholder engagement led to a habitat creation and management scheme that has met its immediate and evidence-based conservation goals in a way which is transferable to a range of different systems.

Objective Analysis of environmental DNA (eDNA) is a method that has been used for the detection of various species within water bodies. The great crested newt (Triturus cristatus) has a short eDNA survey season (mid-April to June). Here we investigate whether this season could be extended into other months using the current methodology as stipulated by Natural England. Results Here we present data to show that in monthly water samples taken from two ponds (March 2014–February 2015) we were able to detect great crested newt DNA in all months in at least one of the ponds. Similar levels of great crested newt eDNA (i.e. highly positive identification) were detected through the months of March–August, suggesting it may be possible to extend the current survey window. In order to determine how applicable these observations are for ponds throughout the rest of the UK, further work in multiple other ponds over multiple seasons is suggested. Nevertheless, the current work clearly demonstrates, in two ponds, the efficacy and reproducibility of eDNA detection for determining the presence of great crested newts. Electronic supplementary material The online version of this article (doi:10.1186/s13104-017-2657-y) contains supplementary material, which is available to authorized users.

Road mitigation tunnels are increasingly deployed for amphibians but very little is known about chemical pollution in such schemes. We assessed pollution pressures associated with road runoff at a major great crested newt mitigation scheme in England. Sediments and waters in the mitigation system were analysed for major physico-chemical parameters, trace metals and total petroleum hydrocarbons and compared to a nearby reference site. Seven out of eight tested metals including copper, zinc, lead and iron were in significantly greater concentrations in the tunnels than at a reference site and at environmentally significant concentrations. Water samples also exhibited elevated concentrations of aluminium and chromium and occasionally extreme alkaline pH associated with leaching of portlandite in tunnel cements. High conductivity values in waters and sediments corresponding with seasonal de-icing salt application were also apparent. The study highlights the potential pollutant pressures for amphibians associated with large-scale urban development and road mitigation schemes.

I Norge er det to salamanderarter, storsalamander (Triturus cristatus) og småsalamander (Lissotriton vulgaris). Dette er det nordligste utbredelsesområdet for salamandere i Europa. Salamandere lever mesteparten av livet sitt på land, men vil dra tilbake til vann når det er parringstid på våren. I denne oppgaven estimerte jeg populasjonsstørrelse og overlevelse for begge artene i en liten gårdsdam i Eidskog i Sørøst-Norge basert på en 4-årig tidsserie. Området rundt dammen er preget av kulturlandskap, myr og skog. I noen tidligere år har dammen tørket inn. Jeg benyttet meg av fangst-gjenfangst metoden, og brukte ruser for å fange salamanderne i seks uker i månedene april til juni. Jeg brukte programmet Amphident for å identifisere individer basert på individuelle bukmønstre. Jeg tok bilder av bukmønsteret på salamanderne, og sammenlignet det med individer fra tidligere fangster. Jeg analyserte dataene jeg samlet inn i MARK, som er et merking-gjenfangstprogram under lukket populasjonsforventning. Mine resultater viste at det var en forskjell i gjennomsnittlig overlevelse mellom storsalamander (74%) og småsalamander (42%) over 4-år. Det var derimot ingen tydelig forskjell i overlevelse mellom kjønn for storsalamander, og heller ikke for småsalamander. Det var derimot en forskjell i overlevelse hos hunnene hos både storsalamander (79%) og småsalamander (37%). Populasjonsestimatet viser at bestanden til storsalamanderne sakte øker fra år til år, (76 til 112 fra 2016 til 2019 for hunner, og 75 til 124 for hanner). Småsalamandernes populasjonsutvikling viser også en svak økning (234 til 262 for hunner, og 271 til 332 for hanner), bortsett fra i året 2018, hvor dataene viser et høyere estimat. Trolig har ikke tørken påvirket salamanderpopulasjonen i noen nevneverdig grad, men man kan kanskje se en forsinket effekt av en manglende årsklasse i senere år. Salamanderes levealder gjør at de kan avvente med reproduksjon til forholdene igjen ligger til rette. Skulle det derimot være tørke i flere år, er det rimelig å anta at man kan se tydelige negative effekter hos begge artene. Fra et forvaltningsperspektiv er det viktig med kunnskap om dammene i nærområdet, samt oppvekst og overvintringsområder, og konnektiviteten mellom dem. Salamanderpopulasjonene er trolig stabile, og har en god overlevelse. Jeg anbefaler at området rundt dammen blir fortsatt ivaretatt, og at arealinngrep som flatehogst ikke gjennomføres da det er oppvekst- og overvintringsområde for salamandere. Jeg foreslår også at man kan gjøre tiltak for å hindre at dammen tørker inn. In Norway, there are two salamander species, great crested newt (Triturus cristatus) and smooth newt (Lissotriton vulgaris). This is the northernmost distribution area for salamanders in Europe. Salamanders live most of their lives on land but will return to water during the breeding season in the spring. In this study, I estimated population size and survival for both species in a small farm pond in Eidskog in southeast Norway based on a 4-year time series. The area around the pond is characterized by cultural landscapes, marshes, and forests. In some previous years, the pond has dried up. I used the mark-recapture method, and used funnel traps to catch the salamanders for six weeks in the months of April to June. I used the Amphident program to identify individuals based on individual abdominal patterns. I took pictures of the abdominal pattern of the salamanders and compared it to individuals from previous catches. I analyzed the data I collected in MARK, which is a mark-recapture program under the expectation that the population was closed. My results showed that there was a difference in average survival between great crested newts (74%) and smooth newts (42%) over 4 years. There was no clear difference in survival between the sexes for great crested newts, nor for smooth newts. On the other hand, there was a difference in the survival of females in both great crested newts (79%) and smooth newts (37%). The population estimate shows that the population of the great crested newt is slowly increasing year by year, (76 to 112 from 2016 to 2019 for females, and 75 to 124 for males). The population of smooth newts also shows a slight increase (234 to 262 for females, and 271 to 332 for males), except in the year 2018, where the data show a higher estimate. Probably, the drought has not affected the salamander population to any significant degree, but one may see a delayed effect of a lack of cohort in later years. Salamanders' longevity means that they can await reproduction until conditions are right again. However, should it be drought for several years, it is reasonable to assume that one can clearly see negative effects in both species. From a management perspective, it is important to have knowledge of the ponds in the immediate area, as well as growing up and wintering areas, and the connectivity between them. Salamander populations are probably stable and have a good survival rate. I recommend that the area around the pond is still maintained and that area intervention such as clear-cuts is not carried out as it is a nursery and hibernation area for salamanders. I also suggest that measures can be taken to prevent the pond from drying in.

Identifying the drivers of population fluctuations in spatially distinct populations remains a significant challenge for ecologists. Whereas regional climatic factors may generate population synchrony (i.e., Moran effect), local factors including the level of density-dependence may reduce the level of synchrony. Although divergences in the scaling of population synchrony and spatial environmental variation have been observed, the regulatory factors that underlie such mismatches are poorly understood.No previous studies have investigated how density-dependent processes and population-specific responses to weather variation influence spatial synchrony at both local and continental scales. We addressed this issue in a pond-breeding amphibian, the great crested newt (Triturus cristatus). We used capture-recapture data collected through long-term surveys in five T. cristatus populations in Western Europe.We found a low level of demographic synchrony at both local and continental levels. Weather has weak and spatially variable effects on survival, recruitment and population growth rate. In contrast, density-dependence was a common phenomenon (at least for population growth) in almost all populations and subpopulations.Our findings support the idea that Moran’s effect is low in species where the population dynamics more closely depends on local factors (e.g. population density and habitat characteristics) than on large-scale environmental fluctuation (e.g. regional climatic variation). Those demographic feature likely have far-reaching consequences for the long-term viability of the spatially structured populations and their ability to response to large-scale climatic anomalies.

As the biggest inner city national reserve in Germany, the D¨onche in Kassel is characterized by its history and composition of ponds, reeds and varying woodlands. Due to artificial grazing, the structural open landscape with small-area wetland habitats was well preserved. In the last two decades, changes in aquatic ecosystems were becoming more obvious. From originally 400 water filled bomb crater only five are left and while several dry out in late spring or early summer, only three might keep enough water for amphibians to reproduce. After the project group “Amphibien- und Reptilienschutz in Hessen e.v.” (Agar) verified the existence of several amphibian species in the D¨onche in 2002, no further detailed inventory on Salamandridae biodiversity and especially the northern crested newt had been conducted. Because of the long time since any latest survey and an obviously changing situation in the water household, we collected information on populations and species diversity for an inventory covering the entire breeding season between March and July.We followed movements of the great crested newt (Triturus cristatus) using a potentially novel tool for newt tracking using body patterns and analyzed the situation of the common newt (Lissotriton vulgaris) while attempting to get an overview on all present amphibian species with a focus on the water quality and water-cycle. As a result of our study we are stating the worrying condition of local amphibian populations due to a major decline in habitat suitability for all aquatic amphibians in the nature reserve D¨onche, most likely caused by an increase in global temperature.

Habitat loss and fragmentation due to urbanisation and road developments have substantial impacts on amphibian populations and road mitigation measures are increasingly used in order to reduce such risks. However, numerous important questions remain unanswered on the actual effectiveness of such ecological road mitigation projects. We used specially adapted time-lapse recording cameras and a custom image analysis script to monitor the amphibian usage and effectiveness of a multi-tunnel mitigation site in northern England over 4 years and used before-and-after pond survey data to evaluate the mitigation results from a population trend perspective. We monitored four amphibian species, including the European protected great crested newt (Triturus cristatus), the main target of amphibian road mitigation in the UK. Despite substantial evidence of mitigation rejection and U-turn movements inside the tunnels, overall, most amphibians entering tunnels successfully used them to move between the different parts of the site separated by the road. Road fences appeared effective for preventing amphibian road mortality and site connectivity was re-established via the wildlife passage system, with multiple new ponds across the road colonised by amphibians. Unlike frogs and toads, tunnel usage by newts was strongly autumn based, indicating that such systems are mainly suitable for supporting newt dispersal movements between breeding areas. The great crested newt population increased rapidly over the course of the monitoring period, suggesting that, together with habitat replacements and improvements, the road mitigation measures were effective at maintaining site connectivity and as a population conservation measure.

Tracking individual animals with small-sized passive integrated transponder tags (PIT tags) has become a popular and widespread method, one which can be used for investigating life history traits, including dispersal patterns of small protected animals such as newts. In this study, we tested the applicability of PIT tag usage for individual marking with the Great crested newt (Triturus cristatus) as a model amphibian species, and to test the detection of the newts in nature using a passive telemetry system. Clove oil was used as an anaesthetic before surgery. We implanted PIT tags under the skin of 140 newts. The survival rate of newts was 98.57%. X-ray images were taken to check the exact positions of the PIT tags. Since approximately 15.71% of the newts were capable of expelling the tag from their bodies, tag loss has to be accounted for in future behavioural studies dealing with newts and other amphibians potentially capable of frequent tag expulsion. Lastly, we detected by passive telemetry 97 individuals out of 100 released into a natural breeding pond. Males had higher activity (13 detected males vs 7 females per hour) than females, thus males could be detected if present with more certainty. The result of the movement behaviour showed that e.g. the male of T. cristatus in a breeding pond can travel up to 20 m in 78 seconds. In summary, this promising method could allow the automatic data collection of marked newts in aquatic as well as in terrestrial biotopes, providing data on their dispersal, diurnal activity and movement behaviour.

In human-modified landscapes, little is known about the influence of aquatic habitat types on the demographic structure of residing amphibian populations. In the present paper, we focus on a European flagship urodele species (the great crested newt Triturus cristatus) at the north-western range of its distribution, applying the method of skeletochronology to compare the ages of individuals retrieved from agricultural ponds with individuals retrieved from aquatic sites favourably managed for T. cristatus presence. Median ages ranged between 4.5 and 10.0 years depending on sex and population, and did not differ between the two site categories. Females were on average older than males at both agricultural ponds as well as favourably managed sites. Median ages at sexual maturity (3 years for females and 2 years for males) were 4 years below the most commonly observed age cohort in both sexes, suggesting that young adults regularly forgo reproduction. Mean body size did not differ between agricultural ponds and favourably managed sites. However, the former were characterised by a higher variance in body size, which is possibly linked to more unstable ecological conditions in agricultural settings. Taken together, our findings confirm that under suitable conditions agricultural ponds can harbour sustainable populations, an important finding for the broad-scale conservation management of T. cristatus which does not usually take population demographies into account.\ud Keywords: Demography Great crested newt Skeletochronology Triturus cristatus Urodeles

International audience; 1. Understanding the mechanisms that regulate the dynamics of spatially structured populations (SSP) is a critical challenge for ecologists and conservation managers. Internal population processes such as births and deaths occur at a local level, while external processes such as dispersal take place at an inter-population level. At both levels, density dependence is expected to play a critical role. At a patch scale, demographic traits (e.g., survival, breeding success) and the population growth rate can be influenced by density either negatively (e.g., competition effect) or positively (e.g., Allee effects). At the scale of an SSP, although positive density-dependent dispersal has been widely reported, an increasing number of studies have highlighted negative density-dependent dispersal. 2. While many studies have investigated the effects of density on population growth or on dispersal, few have simultaneously examined density-dependent effects at the scale of both the local population and the entire SSP. In this study, we examine how density is related to demographic processes at both the pond level (survival and population growth) and the SSP level (between-pond dispersal) in a pondbreeding amphibian, the great crested newt (Triturus cristatus). The study was based on 20 years of individual capture–recapture (CR) data (from 1996 to 2015) gathered from an SSP made up of 12 experimental ponds (“patches”). 3. We first used a CR multievent model to estimate both survival and dispersal rates in specific ponds as a function of distance between ponds. Then, using a second CR multievent model, we examined whether survival and recapture rates were influenced by population density in a pond. Lastly, we used state-space time series models to investigate whether density affected population growth in each pond. 4. Our results found a positive density-dependent effect on survival and a negative density-dependent effect on departure. In addition, the findings indicate that population growth was negatively related to density in all 12 ponds. 5. These results support the hypothesis that in SSPs, density may have multiple and contrasting effects on demographic parameters and growth rates within local populations as well as on dispersal. This study underlines the need to better understand how density dependence may influence potential trade-offs between life-history strategies and life-history stages.

Taugbøl, A., Dervo, B.K., Brandsegg, H. & Fossøy, F. 2019. Analyser av miljø-DNA og strykeprøver for overvåking av soppen Batrachochytrium dendrobatidis (Bd) i Akershus. NINA Rapport 1660. Norsk institutt for naturforskning. Analyser av miljø-DNA er en forholdsvis ny metode for overvåking av arter og økosystemer der innsamling av prøver ikke er avhengig av langvarig innsats eller taksonomisk ekspertise i felt. Denne metoden har vist seg å være svært effektiv med tanke på overvåking av truede arter og arter som det er vanskelig å oppdage visuelt i felt. En slik art er soppen Batrachochytrium dendrobatidis (Bd), som parasitterer huden til amfibier og som kan gi utfall i infeksjonssykdommen chytridiomykose. Det ble for første gang påvist Bd i Norge i 2017, og patogenet ble påvist i fem av femten ulike amfibiedammer i Akershus, sør for Oslo, ved hjelp av miljø-DNA-analyser. I denne rapporten oppsummerer vi prøveinnsamling og analyser for 2018. I tillegg til miljø-DNA-prøver ble det også fange dyr og tatt strykeprøver fra huden. Det ble samlet inn vann fra de 5 kjente infiserte dammene og fem kontrolldammer ved fem ulike tidspunkt, med en forventning om økt utslag av Bd utover sesongen basert på resultater fra 2017. Resultatene fra 2018 viser derimot en motsatt effekt, med lavere konsentrasjoner av Bd inn mot de varmere sommermånedene i de to prøvene som er positive fra samme dam (Østre Støkken) og ingen positive utslag fra den siste innsamlingsrunden. Det ble påvist Bd i en ny dam med vannprøver for 2018, i Nordre Holstad, mens Garderenga ikke fikk utslag på Bd i 2018. Det ble også påvist smitte på dyr i 4 av 5 dammer da ingen av de innfangede dyrene slo positivt ut på Bd i Garderenga. Positive prøver er generelt pålitelige med metoden vi har brukt i denne analysen, og vår erfaring tilsier at falske positiver (påvisning av en art selv om det ikke finnes i lokaliteten) er svært sjeldne. Usikkerheten rundt en negativ prøve er derimot mer uviss, og det at Bd ikke ble påvist i miljø-DNA prøvene selv om strykeprøver hadde positive utslag kan skyldes flere årsaker, som for eksempel vannkvalitet i lokaliteten, temperatur, tetthet av arten, prøvevolumet som ble innsamlet samt behandling og analysering av prøven på lab. En negativ miljø-DNA-prøve bør derfor ikke sees på som et endelig bevis for arten ikke finnes i lokaliteten. Med tanke på de spesielt høye temperaturforholdene sommeren 2018 og lite kunnskap om hvordan livssyklusen til Bd opptrer i våre nordlige dammer, er det gode grunner til å tro at vi enten kan ha samlet inn fra dammene på et ugunstig tidspunkt på dagen, at en forhøyet damtemperatur kan ha ført til atferdsendringer hos vertsdyrene slik at vi ikke fanger opp sporstoffer fra huden i like stor grad som i 2017, eller at syke dyr har hatt kortere oppholdstid i dammene i 2018.

International audience; Dispersal is one of the main processes that determine community structure. Individuals make dispersal decisions according to environmental and/or social cues that reflect the fitness prospects in a given patch. The presence and abundance of heterospecifics within the same ecological guild, and/or their breeding success, may act as public information that influences movement decisions. To date, most studies investigating the role of heterospecific attraction have focused on habitat choice, using both experimental and correlational approaches. The present study is the first to examine how long-term variation in heterospecific density in breeding patches may affect dispersal patterns in spatially structured populations. We investigate how the dispersal decisions of the great crested newt (Triturus cristatus) are related to the variable density of two other newt species, the alpine newt (Ichthyosaura alpestris) and the palmate newt (Lissotriton helveticus). To examine this issue, we used capture–recapture data collected in an experimental pond network over a 20-year period. The results revealed that the great crested newt’s dispersal is context dependent and is affected by variation in heterospecific density: individuals were less likely to emigrate from ponds with high heterospecific density and were more likely to immigrate to ponds with high heterospecific density. These findings suggest that individuals adjust their dispersal decisions at least partly based on public information provided by heterospecifics. This mechanism may play a critical role in the dynamics of spatially structured populations and community functioning.

International audience; Dispersal (i.e. movement from a natal or breeding site to another breeding site) is a central process in ecology and evolution as it affects the eco-evolutionary dynam- ics of spatially structured populations. Dispersal evolution is regulated by the bal- ance between costs and benefits, which is influenced by the individual phenotype (i.e. phenotype-dependent dispersal) and environmental factors (i.e. condition-dependent dispersal). Even though these processes have been extensively studied in species with simple life cycles, our knowledge about these mechanisms in organisms displaying complex life cycles remains fragmentary. In fact, little is specifically known about how the interplay between individual and environmental factors may lead to alternative dispersal strategies that, in turn, lead to the coexistence of contrasted site fidelity phe- notypes. In this paper, we examined breeding dispersal in a pond-breeding amphibian, the great crested newt Triturus cristatus, within usual walking distances for a newt. We took advantage of recent developments in multi-event capture–recapture models and used capture–recapture data (946 newts marked) collected in a spatially structured population occupying a large pond network (73 ponds). We showed a high rate of breeding site infidelity (i.e. pond use) and the coexistence of two dispersal phenotypes, namely, a highly pond faithful phenotype and a dispersing phenotype. Individuals that were site faithful at time t – 1 were therefore more likely to remain site faithful at time t. Our results also demonstrated that the probability that individuals belong to one or the other dispersal phenotypes depended on environmental and individual factors. In particular, we highlighted the existence of a dispersal syndrome implying a covariation pattern among dispersal behavior, body size, and survival. Our work opens new research prospects in the evolution of dispersal in organisms displaying complex life cycles and raises interesting questions about the evolutionary pathways that contribute to the diversification of movement strategies in the wild.

Intensive forest management and landscape degradation are threats to amphibian populations. We modelled and compared the extinction and colonization dynamics of the great crested newt in four different spatial contexts that describe landscape change from past to present and future forest landscapes in eastern Finland. In future landscape scenarios, we explored the effects of two forest use intensities with different logging rotation times. The introduction of fish into breeding ponds has been the main cause of local extinctions of the great crested newt. In the future, intensifying land-use and shorter logging rotation will decrease the connectivity between ponds the most. In conservation planning, more attention should be paid to the spatial arrangement of habitat patches, especially if the intensity of human impact will increase. Conservation effort must be targeted towards securing source ponds in the core area and towards enhancing connectivity.

Recreational activities have become a major use of forests but have also been widely cast as having a negative impact on wildlife and related nature conservation. Natural science analyses dominate the study of the relationship between recreation and wildlife, and yet, there are also critical social scientific dimensions. These include understanding the beliefs that recreationists hold regarding how their activities impact on wild species and their habitats. These beliefs are important because individual understandings of the consequences of behaviours are closely related to how they behave in practice. This paper presents a study of the perceptions of forest visitors concerning recreational impacts on amphibians, such as the great crested newt (Triturus cristatus). Amphibians have been neglected in the recreational disturbance literature despite posing some of the most substantial conservation challenges. In this study, data generated by a face-to-face survey of visitors at two forest sites in Scotland reveals that nearly half of recreationists surveyed did not believe that their activities had much impact on amphibians but pointed to other activities as being responsible for any potential issues. Recreationists who visit forests the most felt that they had the least impact. Despite this, management methods to reduce recreational disturbance are broadly supported. Respondents also indicated that they favoured forms of communication such as codes of conduct and educational programmes, as ways to raise awareness and learn about this issue. We conclude by reflecting on the implications of our results for forest managers who may wish to engage visitors and minimise the disturbance of wildlife.

The use of environmental DNA (eDNA) is rapidly emerging as a potentially valuable survey technique for rare or hard to survey freshwater organisms. For the great crested newt (Triturus cristatus) in the UK, the substantial cost and manpower requirements of traditional survey methods have hampered attempts to assess the status of the species. We tested whether eDNA could provide the basis for a national citizen science-based monitoring programme for great crested newts by (i) comparing the effectiveness of eDNA monitoring with torch counts, bottle trapping and egg searches and (ii) assessing the ability of volunteers to collect eDNA samples throughout the newt’s UK range. In 35 ponds visited four times through the breeding season, eDNA detected newts on 139 out of 140 visits, a 99.3% detection rate. Bottle traps, torch counts and egg searches were significantly less effective, detecting newts 76%, 75% and 44% of the time. eDNA was less successful at predicting newt abundance being positively, but weakly, correlated with counts of the number of newts. Volunteers successfully collected eDNA samples across the UK with 219 of 239 sites (91.3%) correctly identified as supporting newts. 8.7% of sites generated false negatives, either because of very small newt populations or practical difficulties in sample collection. There were no false positives. Overall, we conclude that eDNA is a highly effective survey method and could be used as the basis for a national great crested newt monitoring programme.

Since its introduction in 1974, the use of glyphosate in agriculture has been continuously increasing; however, the application modes of this herbicide have been changing. Therefore, glyphosate-based herbicides can be used as an appropriate indicator for assessing how changes in pesticide application modes affect wild-living organisms in agricultural landscapes over time. Amphibians that migrate through arable fields may be exposed to the chemicals applied to field crops. Using data on the tem-poral coincidence of four amphibian populations with glyphosate applications from a three-year in-vestigation in northeast Germany as well as data on the application of glyphosate to field crops in German agriculture over 20 years, we estimated the species-specific increasing rates of coincidence likelihoods during this period. The overall consumption of glyphosate used in German agriculture between 1992 and 2012 increased by a factor of 5.7, while the species-specific coincidence likelihood increased from 2.2 to 6.1, respectively. Our results reveal the highest increases in coincidence for both adult and juvenile great crested newt (Triturus cristatus) and fire-bellied toad (Bombina bombina). Adults and juveniles of moor frog (Rana arvalis) and adults of spadefoot toad (Pelobates fuscus) were subjected to moderate increases, with rates ranging from 3.2 to 3.6; in contrast, juvenile individuals of P. fuscus showed small increases. We suggest that the risk assessments of pesticide application (in this case, glyphosate) should not only consider the present use at the time of authorization but also consider changes in application modes over time that may lead to increases in potential exposure of non-target organisms, such as amphibians.

The use of aquatic environmental DNA (eDNA) to detect the presence of species depends on the seasonal activity of the species in the sampled habitat. eDNA may persist in sediments for longer than it does in water, and analysing sediment could potentially extend the seasonal window for species assessment. Using the great crested newt as a model, we compare how detection probability changes across the seasons in eDNA samples collected from both pond water and pond sediments. Detection of both aquatic and sedimentary eDNA varied through the year, peaking in the summer (July), with its lowest point in the winter (January): in all seasons, detection probability of eDNA from water exceeded that from sediment. Detection probability of eDNA also varied between study areas, and according to great crested newt habitat suitability and sediment type. As aquatic and sedimentary eDNA show the same seasonal fluctuations, the patterns observed in both sample types likely reflect current or recent presence of the target species. However, given the low detection probabilities found in the autumn and winter we would not recommend using either aquatic or sedimentary eDNA for year-round sampling without further refinement and testing of the methods.

Taugbøl, A., Dervo, B.K., Brandsegg, H., Fossøy, F. 2018. Analyser av miljø-DNA for påvisning av soppen Batrachochytrium dendrobatidis (Bd) i Østfold. NINA Rapport 1564, Norsk institutt for naturforskning. Analyser av miljø-DNA er en forholdsvis ny metode for overvåking av arter og økosystemer der innsamling av prøver ikke er avhengig av langvarig innsats eller taksonomisk ekspertise i felt. Denne metoden har vist seg å være svært effektiv med tanke på overvåking av truede arter og arter som det er vanskelig å oppdage visuelt i felt. En slik art er soppen Batrachochytrium dendrobatidis (Bd), som parasitterer huden til amfibier og som kan gi utfall i infeksjonssykdommen chytridiomykose. Denne soppen ble påvist i vannprøver fra fem lokaliteter i Akershus i 2017 og det var ønskelig å kartlegge et større område for å se på utbredelse av Bd. Det ble derfor samlet inn vannprøver fra 25 lokaliteter i Østfold, der prøvene ble testet for tilstedeværelse av Bd, storsalamander og småsalamander. Samtlige prøver var negative for Bd, 16 dammer var positive for storsalamander og 21 dammer var positive for småsalamander. Gjennomsnittlig antall DNA kopier per mL med standardavvik for storsalamander var 5,397 ± 14,11, mens det for småsalamander var 2,884 ± 4,66. Positive prøver er generelt pålitelige med metoden vi har brukt i denne analysen, og vår erfaring tilsier at falske positiver (påvisning av en art selv om det ikke finnes i lokaliteten) er svært sjeldne. Usikkerheten rundt en negativ prøve er derimot mer uviss, og det at Bd ikke ble påvist i noen av dammene kan skyldes flere årsaker, som for eksempel vannkvalitet i lokaliteten, temperatur, tetthet av arten, prøvevolumet som ble innsamlet samt behandling og analysering av prøven på lab. En negativ miljø-DNA-prøve bør derfor ikke sees på som et endelig bevis for at arten ikke finnes i lokaliteten. Med tanke på de spesielt varme temperaturforholdene sommeren 2018 og en noe manglende kunnskap om livssyklusen til Bd i våre nordlige dammer, er det gode grunner til å tro at vi enten kan ha hentet inn prøver fra dammene på et ugunstig tidspunkt, at en forhøyet vanntemperatur har ført til atferdsendringer hos vertsdyrene slik at vi ikke fanger opp sporstoffer fra huden i like stor grad som i 2017, eller at syke dyr i mindre grad oppholder seg i dammene utover sommeren. Vi kan med andre ord ikke utelukke falske negative resultater for Bd og anbefaler en ny, utvidet kartlegging i 2019. Taugbøl, A., Dervo, B.K., Brandsegg, H., Fossøy, F. 2018. Analyser av miljø-DNA for påvisning av soppen Batrachochytrium dendrobatidis (Bd) i Østfold. NINA Report 1564. Norwegian Institute for Nature Research. Analyses of environmental DNA (e-DNA) is a promising method for surveying species and ecosystems, as the method is cheap and there is no demand of taxonomic expertise in the field. Further, examining environmental samples for DNA has proven efficient with concern to the detection of threatened species and species that are hard to detect in the field. One such species is the invasive fungus Batrachochytrium dendrobatidis (Bd) that parasitizes the skin of many amphibian species, often resulting in a lethal infections disease called chytridiomycosis. Bd was detected in Norway for the first time in five localities 2017 (25% of the investigated ponds in Akershus), and there was a need to extend the sampling area to further improve our knowledge of where Bd was present in Norway. As part of a research project financed by the County Governor, NINA has collected water samples from 25 different ponds in Østfold County for analysis of environmental DNA (eDNA) on Bd, where the results are summarized in this report. All samples were negative for Bd, 16 ponds were positive for northern crested newt (Triturus cristatus) and 21 ponds were positive for common newt (Triturus vulgaris). The average number of DNA-copies per mL with standard error for the northern crested newt was 5,397 ± 14,11, while the same numbers for common newt was 2,884 ± 4,66. That Bd was not detected in any of the ponds could be caused by false negative samples. Several sampling factors could lead to false negative samples, such as the time of sampling in the season and in the day, a potentially changed behaviour of infected animals, temperature in the pond, the amount of DNA leaking from the targeted species and the amount of water that was collected and filtered. Fur-ther, as each pond were only sampled once, we should therefore not treat the negative results as final. Also, with the especially warm summer of 2018, our low knowledge of how Bd’s life cycle is enacting in our northern ponds and our for the most part negative results from the infected ponds from 2017 (unpublished), we believe that some of the ponds indeed are false negatives and suggest a new, extended sampling effort in the same region in 2019.

International audience; Advances in the sequencing of DNA extracted from media such as soil and water offer huge opportunities for biodiversity monitoring and assessment, particularly where the collection or identification of whole organisms is impractical. However, there are myriad methods for the extraction, storage, amplification and sequencing of DNA from environmental samples. To help overcome potential biases that may impede the effective comparison of biodiversity data collected by different researchers, we propose a standardised set of procedures for use on different taxa and sample media, largely based on recent trends in their use. Our recommendations describe important steps for sample pre-processing and include the use of (a) Qiagen DNeasy PowerSoil ® and PowerMax ® kits for extraction of DNA from soil, sediment, faeces and leaf litter; (b) DNeasy PowerSoil ® for extraction of DNA from plant tissue; (c) DNeasy Blood and Tissue kits for extraction of DNA from animal tissue; (d) DNeasy Blood and Tissue kits for extraction of DNA from macroorganisms in water and ice; and (e) DNeasy PowerWater ® kits for extraction of DNA from microorganisms in water and ice. Based on key parameters, including the specificity and inclusivity of the primers for the target sequence, we recommend the use of the following primer pairs to amplify DNA for analysis by Illumina MiSeq DNA sequencing: (a) 515f and 806RB to target bacterial 16S rRNA genes (including regions V3 and V4); (b) #3 and #5RC to target eukaryote 18S rRNA genes (including regions V7 and V8); (c) #3 and #5RC are also recommended for the routine analysis of protist community DNA; (d) ITS6F and ITS7R to target the chromistan ITS1 internal transcribed spacer region; (e) S2F and S3R to target the ITS2 internal transcribed spacer in terrestrial plants; (f) fITS7 or gITS7, and ITS4 to target the fungal ITS2 region; (g) NS31 and AML2 to target glomeromycota 18S rRNA genes; and (h) mICOIintF and jgHCO2198 to target cytochrome c oxidase subunit I (COI) genes in animals. More research is currently required to confirm primers suitable for the selective amplification of DNA from specific vertebrate taxa such as fish. Combined, these recommendations represent a framework for efficient, comprehensive and robust DNA-based investigations of biodiversity, applicable to most taxa and ecosystems. The adoption of standardised protocols for biodiversity assessment and monitoring using DNA extracted from environmental samples will enable more informative comparisons among datasets, generating significant benefits for ecological science and biosecurity applications.

Elgtvedt, I. & Dervo, B. K. 2018. Kartlegging av amfibier i utvalgte ynglelokaliteter i Oslo kommune. NINA Rapport 1574. Norsk institutt for naturforskning. Denne rapporten beskriver resultatene av feltarbeidet som ble utført for å kartlegge forekomsten av småsalamander (Lissotriton vulgaris), storsalamander (Triturus cristatus) og frosk (Rana sp.) i utvalgte lokaliteter i Oslo kommune på oppdrag for Bymiljøetaten i 2018. Totalt ble 15 dammer undersøkt. Alle dammene er tidligere undersøkte ynglelokaliteter med unntak for dammen ved Grønmo golfbane. Informasjonen for mange av dammene er fra eldre feltundersøkelser, og må-let med kartleggingen har vært å oppdatere kunnskapen om disse utvalgte amfibielokalitetene. Det ble fanget voksne småsalamandre i 11 av de 15 dammene, til sammen 946 hanner og 736 hunner. I to av dammene uten fangst av voksne, Grønmo golfbane og Vadedammen, ble det kun funnet larver av småsalamander. Det ble fanget voksne storsalamander i 5 av de 15 dam-mene, til sammen 76 hanner og 91 hunner. I Vadedammen er det i tillegg et ubekreftet funn av larver til storsalamander. Det er behov for å gjennomføre skjøtselstiltak for storsalamander i en eller to av dammene, mens det er behov for å gjennomføre tiltak i 6-7 dammer for småsalamander. Behovet for skjøtsel ser ut til å være størst i Møllesvingen i forhold til storsalamander. For små-salamander synes behov for restaurering å være størst for alle dammene på Jomfrubråtveien, Øvre Ljan, Båntjernveien 5 og Andersendammen. Det ser ut til å være behov for både fjerning av kantvegetasjon, våtmarksvegetasjon og fjerning av noe sedimenter og vannvegetasjon. Elgtvedt, I. & Dervo, B. K. 2018. Survey of amphibians in selected reproduction localities in Oslo municipality. NINA Report 1574. Norwegian Institute for Nature Research. This report describes the fieldwork performed to map the occurrence of smooth newt (Lissotriton vulgaris), great crested newt (Triturus cristatus) and frogs (Rana sp.) in selected localities in Oslo municipality on mission from Bymiljøetaten in 2018. In total were 15 ponds examined. All the selected breeding localities have been examined earlier, except from the pond by Grønmo golf course. The information about many of the ponds are from other field surveys, and the aim of this survey have been to update the knowledge about the selected amphibian localities. There were captured grown smooth newt in 11 of the 15 ponds, in total 946 males and 736 females. In the two ponds where no adults of smooth newt were observed, Grønmo golf course and Vadedammen, there was only found larva. There were captured grown great crested newt in 5 of the 15 ponds, in total 76 males and 91 females. In Vadedammen there was also an unconfirmed number of great crested newt larva. There is a need to carry through management plans for great crested newt in one or two of the ponds, while there is a need to carry out measures in 6-7 of the ponds for smooth newt. The need for management plans are highest in Møllesvingen in relations to great crested newt. For smooth newt, it is a hign need for restoration in all the ponds on Jomfrubråtveien, Øvre Ljan, Bånthernveien 5 and the pond Andersendammen. It looks like there are both needs to remove edge vegetation, wetland vegetation and to remove sediments and aquatic vegetation.

Приведены сведения о встречах в 2017-2018 гг. амфибий и рептилий, занесенных в Красную книгу Свердловской обл., Slowworm Anguis fragilis, Smooth Snake Coronella austriaca, and Great Crested Newt Triturus cristatus were recorded in the Sverdlovsk region in 20172018. All these species are included in the regional Red Data Book.

Taugbøl, A., Dervo, B.K., Brandsegg, H., Sivertsgård, R. Fossøy, F. 2018. Bruk av miljø-DNA til overvåkning av små- og storsalamander. NINA Rapport 1476. Norsk institutt for naturforskning. I denne rapporten oppsummerer vi resultatene fra uttesting av miljø-DNA som en metode for overvåkning av småsalamander og storsalamander i Akershus fylke og Lier kommune. Miljø-DNA er definert som alt DNA som har blitt tilført omgivelsene fra ulike arter, via bl.a. hudrester, spytt og avføring. Ved hjelp av miljø-DNA kan man påvise tilstedeværelse av arter uten direkte observasjon. Resultater fra en tidligere uttesting av miljø-DNA fra salamanderdammer viste stor variasjon i DNA-konsentrasjon mellom punktprøver tatt i samme dam. Det ble derfor testes ut ulike prøve-oppsett for innsamling av vann i to dammer i første testrunde for denne rapporten; forskjellige filter, preserveringsmetoder og vannvolum. Etter sammenligning av disse resultatene konklu-derte vi med å bruke en 0,5 liter vannprøve fra en oppsamlingskanne (totalt 15 liters blandeprøve fra 15 stasjoner) filtrert igjennom 0.45 μm filter preservert i 1400 μl ATLbuffer for oppbevaring frem mot ekstraksjon. For hver dam ble det samlet inn minimum en oppsamlingskanne, som ble filtrert for minimum en prøve. Det ble også samlet inn flere oppsamlingskanner på samme tidspunkt for flere dammer, opptil tre oppsamlingskanner og seks DNA-prøver for et tidspunkt. Vi finner noe variasjon mellom prøver samlet inn fra samme oppsamlingskanne og noe variasjon mellom oppsamlingskanner samlet inn fra samme dam til samme tidspunkt, men totalt sett er det god sammenheng i DNA konsentrasjonen mellom prøvene tatt med denne innsamlingsmetoden. Alle de kjønnsmodne salamanderne oppholder seg i yngledammen ca. to uker etter at isen har gått og er der hovedsakelig frem til midten av juli. Ved å samle inn flere dammer igjennom sesongen kan vi få et innblikk i hvordan DNA konsentrasjonen endrer seg over tid, selv om denne også kan variere pga. variasjon i temperatur og med økt parrings adferd. For begge disse faktorene forventer vi en økning av mengden DNA i vannet. Et utvalg av dammene i denne rapporten ble undersøkt opptil fire ganger. Resultatene indikerer ikke en jevn økning gjennom sesongen, men heller en økning før en reduksjon, så en ny økning. Det trengs mer utprøving før resultatene viser at vi kan konkludere med at dette er en trend eller en ren tilfeldighet. Småsalamander har høyere konsentrasjon av miljø-DNA enn storsalamander i denne undersøkelsen, og dette er også reflektert i fangst per innsats (CPUE) for begge områdene. Selv om det er en forholdsvis god sammenheng mellom mengdeforholdet av de to artenes miljø-DNA og fangstdata for de ulike dammene, er det usikkerheter rundt hvordan vi bør omregne den lavere kroppsstørrelsen til småsalamander og mengden av dennes miljø-DNA. Vi konkluderer med at miljø-DNA er en svært godt egnet metode for påvisning av amfibier. Ved å videreutvikle metoden er det stor grunn til å tro at miljø-DNA vil kunne være en kostnadseffektiv metode for påvisning av samtlige amfibiearter, og metoden vil også kunne gi indikasjoner på bestandsstørrelsen til de ulike artene i dammene som prøvetas. Taugbøl, A., Dervo, B.K., Brandsegg, H., Sivertsgård, R. Fossøy, F. 2018. Bruk av miljø-DNA til overvåkning av små- og storsalamander. NINA Rapport 1476. Norsk institutt for naturforskning. This rapport summarizes the findings of using environmental DNA (eDNA) as a tool for monitoring the two present newts in Norway, the common newt and the crested newt. eDNA is all the genetic material obtained directly from environmental samples without necessarily direct observation or sampling of the species. The total amount of DNA shredded from animals present in the environment, through e.g. skin, saliva and feces, is isolated, and the targeted species is amplified with species-specific primers. The samples collected for this report were water samples from two municipalities in Norway, Akershus and Buskerud. Results from earlier testing of eDNA in water ponds inhabited by newts identified a large variation in DNA abundance between samples collected from the same pond. We wanted to decrease the variation between samples taken from the same pond, so we first tested a variety of water volume, filters and storage-methods of the filters in the first field sampling in 2017. This resulted in a standardized sampling method of first collecting 1 L of water at 15 sites spread across a certain distance, we then sampled two replicated samples of 0,5 L water from the 15 L batch for filtration, the water was filtrated through a 0.45 μm filter and stored in 1400 μl ATL buffer until the filter was processed in the lab. We sampled a total of 30 ponds, where many ponds were sampled more than one time. For each pond and sampling time, we collected one-three 15 L batch of water and filtered at least 2 replicated samples from each batch. The results indicate some variation between replicated samples filtered from the same batch, and some variation between samples filtrated from separate batches, but the total variation between replicated samples and batches was low. As all the mature newts are inhabiting the ponds about two weeks after the ice breaks and stays in the ponds until mid-July, we could compare how the eDNA concentration in the ponds changes with factors such as reproductive activity and seasonal temperature, where we expected an increase in the amount of eDNA with increased mating behavior, that again are linked to temperature. As some of the ponds were sampled several times during the season, we could check for any such correlations. The results indicate that there is an increase in the amount of eDNA to-ward late May, until it increases again toward our latest sampling in late June. As we only have a small subset of ponds that were sampled multiple times, we need to check this trend by more careful sampling in the future. There was a higher abundance of eDNA from the common newt compared to the crested newt, and this is also reflected by catch per unit effort data for the same ponds. We do not, however, know if the shredding rate of the two species reflect their size, as this would indicate that the eDNA of the common newt should be about half that of the crested newt. More careful studies are needed in order to quantify species specific differences in eDNA quantity of the two species. This report identifies eDNA as a promising tool for future monitoring, both to identify new ponds with presence/absence, but also to indicate the number of individuals that inhabits each pond.