Evaluation of the Composition of Ichthyofauna in Lakes Free from Commercial Use in a Tightly Protected Area of the Wolin National Park (Poland)

The ichthyological biodiversity of Polish lakes is poorly understood, even in lakes not used for fishing and those under strict protection. With this in mind, the aim of this study was to conduct a comprehensive ichthyological survey of several lakes in the Wolin National Park (Poland). The study was conducted in Domysławskie and Gardno lakes and the anthropogenic Stara Kredownia reservoir. Each fishing exercise was conducted using benthic nets over a 12 h period. The fish caught were measured and weighed, and the species identified. Shannon-Wiener (H) and Simpson biodiversity indices were determined. The catch comprised a total of 4580 fish (of which 4391 were caught in Domysławskie Lake) belonging to 15 species, with a combined weight of 98,165 g. In general, the most prevalent species were non-predatory fish: roach (Rutilus rutilus), freshwater bream (Abramis brama), white bream (Blicca bioerkna), bleak (Alburnus alburnus), and rudd (Scardinius erythrophthalmus). Individually, perch (Perca fluviatilis) and roach (Rutilus rutilus) were the most abundant species. Ichthyofauna diversity varied greatly between the studied water bodies, with 15 species found in Domysławskie Lake and only 6 species in Gardno Lake. In Stara Kredownia Reservoir, no fish were caught, but amphibians were observed, such as great crested newts (Triturus cristatus) and common newts (Lissotriton vulgaris). Based on the results obtained, Lake Domysławskie had a higher species ichthyofauna diversity than the other ‘tench and pike’ lakes. Gardno Lake had a much lower biodiversity.

How reliable is the habitat suitability index as a predictor of great crested newt presence or absence?

The application of a habitat suitability index (HSI) assessment to predict the use of ponds by great crested newts (Triturus cristatus) is commonly used in association with distribution and monitoring projects. Such projects are often used to inform development and planning decision making. However, this type of assessment is frequently misused, and misinterpreted. We used a large, commercially collected environmental DNA (eDNA) survey for great crested newt pond occupancy (489 ponds) to; (1) assess whether it is appropriate to use low HSI scores to rule out occupancy, (2) discuss the use of high HSI scores to identify ponds of high importance for the species and, (3) explore the eDNA detection method. We conclude that there is no evidence to support ruling out pond occupancy based on low HSI scores. However, the conventional view that ponds with HSI scores above 0.7 are of high importance to great crested newts is somewhat supported by the data. Both eDNA and direct observational survey methodologies suffer from sampling error and these need to be acknowledged in the analysis of large data sets.

Establishing Cell Lines from Fresh or Cryopreserved Tissue from the Great Crested Newt (Triturus cristatus): A Preliminary Protocol

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.

Zur Bedeutung der ‚Grünbrücke Kiebitzholm‘ (Schleswig-Holstein) für Amphibien

In den Jahren 2006 bis 2010 wurde die Amphibienfauna auf der Grünbrücke bei Kiebitzholm (Schleswig-Holstein, Norddeutschland), an den angrenzenden Straßensäumen beidseitig der Bundesautobahn (BAB) A21 sowie an Stillgewässern im 1,5 km Radius um die Grünbrücke untersucht. Im Untersuchungsgebiet konn- ten sieben Amphibienarten festgestellt werden, darunter Knoblauchkröte (Pelo- bates fuscus), Kreuzkröte (Epidalea calamita) und Moorfrosch (Rana arvalis), die landes- oder bundesweit als bestandsgefährdet gelten. Bei Knoblauchkröte und Moorfrosch handelt es sich zudem um bundesweit bedeutsame Zielarten für den Biotopverbund. Teichmolch (Lissotriton vulgaris), Erdkröte (Bufo bufo), Grasfrosch (Rana temporaria) und Moorfrosch wurden im Untersuchungszeitraum auf der Grünbrücke nachgewiesen. Der Kammmolch (Triturus cristatus) konnte nur auf der östlichen Rampe festgestellt werden. Die Grünbrücke diente den er- fassten Individuen offenbar in erster Linie zur Querung der BAB A21 und nur zum Teil auch als Sommerhabitat. Eine auf der östlichen Rampe angelegte Wildtränke wurde im Jahr 2010 erstmals von Teichmolch, Kammmolch und Moorfrosch als Laichgewässer genutzt. Knoblauchkröte und Kreuzkröte waren im Untersuchungsgebiet besonders selten. Sie traten in einem bzw. zwei Untersuchungsgewässern auf. Während die Kreuz- kröte auf beiden Seiten der Autobahn gefunden wurde, wurde die Knoblauchkröte nur westlich der Straße erfasst. Die übrigen festgestellten Amphibienarten traten in teilweise größeren Populationen auf beiden Seiten der BAB A21 auf. Die Funde dieser Arten auf der Grünbrücke zeigen, dass bereits ein Verbund zwischen diesem Que- rungsbauwerk und den Populationen in der Umgebung realisiert war.

Ready-to-use workflows for the implementation of genetic tools in conservation management

We present a conservation genetics tool kit, which offers two ready-to-use workflows for the routine application of genetic methods in conservation management. The workflows were optimized for work load and costs and are accompanied by an easy-to-read and richly illustrated manual with guidelines regarding sampling design, sampling of genetic material, necessary permits, laboratory methods, statistical analyses and documentation of results in a practice-oriented way. The manual also provides a detailed interpretation help for the implementation of the results in conservation management. One workflow deals with the identification of pond-breeding amphibians based on metabarcoding and environmental DNA (eDNA) from water samples. This workflow also discriminates the morphologically similar water frogs (Pelophylax sp.) and other closely related species (e.g. Triturus cristatus and T. carnifex). The second workflow studies connectivity among populations using microsatellite markers. Its statistical analyses encompass the detection of genetic groups and historical, recent and current dispersal and gene flow. Using the two workflows does not involve academic research institutes; they can be applied by environmental consultancies, laboratories from the private sector, governmental agencies or non-governmental organisations. These and additional conservation genetic workflows will hopefully foster the routine use of genetic methods in conservation management.

Amphibians in an urban environment: a case study from a central European city (Wrocław, Poland)

In 2016, we studied the occurrence of amphibians in 231 selected ponds in the city of Wrocław (Lower Silesia, Poland) and confirmed the occurrence of 10 species: Bombina bombina, Bufo bufo, Bufotes viridis, Hyla arborea, Pelobates fuscus, Pelophylax esculentus complex, Rana arvalis, Rana temporaria, Lissotriton vulgaris and Triturus cristatus. Among all studied sites, the most common were P. esculentus complex (146 occupied sites, 63.2%) and B. bufo (119, 51.5%), while the rarest was P. fuscus (8, 3.5%). The number of species at a single site varied from 0 (28 sites, 12.1%) to 9 (1 site, 0.4%). Frequency and mean amphibian species richness reached 87.9% and 2.7 ± 1.9, respectively. The composition of amphibian species did not change in comparison to previous studies conducted in the years 1997–2009, but declining trends in five species (B. bombina, B. viridis, H. arborea, P. esculentus complex, P. fuscus) were observed (28 compared sites). The results revealed that the permanency of ponds, their occurrence in the vicinity of river valleys, and a high ratio of watercourse length and green area around ponds are positively correlated and have a significant influence on amphibian species richness within the city. Thus, these identified factors should be considered in the course of sustainable urban planning in order to avoid potential conflicts between nature conservation and urban development.