Effects of Acute Ammonium Nitrate Levels Caused by Agricultural Activities on Four Amphibian Species in The Eastern Black Sea Region

The tadpoles of four amphibian species, namely the Marsh Frog (Pelophylax ridibundus), the Iranian Long-Legged Frog (Rana macrocnemis), the Caucasian Parsley Frog (Pelodytes caucasicus) and the Variable Green Toad (Bufotes variabilis), were exposed to acute concentrations (0 to 500 mg/L) of ammonium nitrate to assess the lethal effects (larval growth, abnormalities, mortality, and LC50 values). Eggs of each species were obtained from clean and polluted habitats in the same region and the tadpoles for experiments were provided from those eggs in the laboratory conditions. Although there was some variability between different populations of the same species or between different species in the observed effects, acute levels of ammonium nitrate caused decreased growth rate and increased abnormalities and mortality in general. Among the 4 amphibian species, the Variable Green Toad was the most damaged one in terms of growth reduction (on average 77-83%), and abnormality rates, and the most damaged one in terms of mortality rates was the Marsh Frog (on average 61-72%). Additionally, the species with the lowest concentration of ammonium nitrate, which killed half of its population, was the Marsh Frog. LC50 values for two populations of Marsh Frog were 37 and 59 mg/L. As a result of our research, it was determined that the acute fertilizer levels caused by agricultural activities in the region had very important harmful effects for all the species we examined. In this context, it can be said that very important environmental and biodiversity problems may occur if certain precautions are not taken regarding the use of the fertilizers and if the awareness of the farmers using these fertilizers cannot be raised.

Growth Layers and Its Complex Structure in a Common Species Under Uncommon Conditions: Pelophylax ridibundus in the Talysh Mountains

In the Talysh Mountains, the marsh frog, Pelophylax ridibundus (Pallas, 1771), increases its distribution due to human activity. In the mountain forest belt, frogs inhabit flowing ponds with cold spring water. These reservoirs are characterized by a stable temperature regime: the water in them is kept at the level of 10 – 12°C in winter and does not fall below 6°C, and in summer does not rise above 18°C. Probably, the cold flow water of ponds in the mountain-forest belt of Talysh can cause some features of growth and maturation for P. ridibundus. The aim of our work was to study the features of the structure of growth layers including the variation in degree of expression of lines of arrested growth (LAGs) of the marsh frogs in ponds with cold flow water. Frogs were collected in the upper part of the Tangeru River gorge in Sym village of Astara District of Azerbaijan (480 m a.s.l.) in August 2018. In total, we studied skeletochronologically 8 females and 9 males. In contrast to the results of the tubular bones studies in the marsh frog presented in earlier works, the studied individuals are characterized by a more complex and diverse structure in the cross section of shin bones. All the revealed diversity of this structure can be divided into three groups. To the first group are relatively rare cases in which there are growth layers with wintering LAGs and additional growth layers but without additional LAGs. The second group includes more frequent cases in which there are growth layers not only with wintering LAGs but with additional LAGs differing discretely from wintering LAGs. The third group includes the rarest cases where there are growth layers with wintering LAGs and with additional LAGs without discrete difference from wintering LAGs. The revealed high variable structure of growth layers and distinctiveness of LAGs in P. ridibundus can be explained by unusual temperature regime in habitat of studied population and the existence of several activity period during a year. The most continuous period of low temperatures (February and March) corresponds to formation hibernation LAGs and dark rings in growth layers. Besides, period from early December to late January may be so cold that frog growth retard sufficiently or stop completely. The formation of more dark parts of growth layers or even several additional LAGs can correspond to this period.

Faunistic review of amphibians in protected areas of Ukrainian Roztochia

Background. Study of amphibian fauna as a necessary component of ecosystems is always relevant and requires a regular renewal of information, especially in protected areas. We conducted a field investigation of amphibians, a literature review and an inventory of the collections of herpetological funds in relation to three protected areas: the Roztochia Biosphere Reserve, the Yavorivskyi National Park, and the Cholgini Ornithological Reserve. Materials and Methods. The field study was performed by classical methods: route method with manual catching and with herpetological nets, vocalization method, sampling of amphibians crushed on the road, and catching individuals with frog fences. We also analyzed the literature and amphibian collections in the Zoological Museum of Ivan Franko National University of Lviv. Results. According to the available literature, the amphibian fauna of these three areas under protection is similar and typical of this region. The results of our field research slightly differ from the literature data, thus, there is a need for further studies. On the study areas, we found 12 species of amphibians, which is more than half of the number of amphibian species in Ukraine, in particular: Smooth Newt Lissotriton vulgaris, Great Cres­ted Newt Triturus cristatus, Fire-bellied Toad Bombina bombina, Eastern Tree Frog Hyla orientalis, Common Spadefoot Toad Pelobates fuscus, Common Toad Bufo bufo, Green Toad Bufotes viridis, Common Frog Rana temporaria, Moor Frog Rana arvalis, Marsh Frog Pelophylax ridibundus, Edible Frog Pelophylax esculentus and Pool Frog Pelophylax lessonae. We detected 2 species of Caudata and 10 species of Anura. 10 species of amphibians were found in the Roztochia Biosphere Reserve, 5 species – in the Yavorivskyi National Park, and 9 species – in the Cholgini Ornithological Reserve. Conclusions. According to the field results, 4 species inhabit all three protected areas – Common Toad, Eastern Tree Frog, Marsh Frog and Edible Frog. A relatively small variety of amphibians found in the Yavorivskyi National Park can be due to the homogeneity of habitats and a proximity of one of the largest military proving grounds in Europe, which limits amphibians in choice of the reproductive and trophic habitats. These habitats deserve special attention because of their fast degradation for the last 10 years. Keywords: amphibians, fauna, protected areas, Ukrainian Roztochia, the Roztochia Biosphere Reserve, the Yavorivskyi National Park, the Cholgini Ornithological Reserve