Testate Amoebae in Karst Caves of the Dinaric Arc (South-Eastern Europe) with a Description of Centropyxis bipilata sp. nov.

Protists in caves are scarcely researched. Most cave studies address the diversity of protists, but very little is known about their habitats and spatio-temporal dynamics. The aim of this study was to investigate the diversity and abundance of protists in the Veternica cave in the Medvednica mountain in Croatia on hygropetric and sinter and clay pools during six months. During this study, 47 protists taxa were distinguished belonging to the groups of heterotrophic flagellates, ciliates, heliozoans, stramenopiles and testate and naked amoebae. The highest taxon richness was found at the sinter pool site richest in bats guano (36 taxa). Most diverse were testate amoebae and ciliates. The number of taxa and their abundance differed significantly between the sampling sites. The prospected habitats supported different protist assemblages, proved by ordination analysis. The most distinctive habitat was the hygropetric. Habitat heterogeneity could be attributed to the presence of inorganic and organic sediments at the sites and habitat microhydrology (lotic or lentic system). Kendall’s concordance coefficient showed a good synchronicity between the habitats in the Veternica cave, based on taxon richness and abundance of protists indicating similar seasonal trends. Seasonality in the studied habitats is attributed to the hydro-meteorological conditions in the Veternica cave drainage area. This study is one of the few studies of spatio-temporal diversity and abundance of protists in caves. Despite the similar appearance of small transitional and aquatic habitats in caves, an example of this study showed specific habitat diversification.

Microphototrophs (algae and cyanobacteria) in karst environments have been intensively studied in aquatic epigean habitats. In recent decades knowledge about the communities inhabiting cave entrances and lampenflora has grown substantially, but the data about the communities in aphotic cave zone are scarce. This study aimed to investigate spatio-temporal presence of microphototrophs in the aphotic zone of Veternica Cave (Mt. Medvednica karst) and to present additional preliminary data from 22 caves of the Dinaric karst. The data were collected over ten years, in parallel with research on cave phagotrophic protists. In addition to the remains of microphototrophs, living algae and cyanobacteria were found in the cave aphotic zone. Diatoms (Bacillariophyta) were the most frequent group found, followed by green algae (Chlorophyta), golden-brown algae (Chrysophyta) and the filamentous cyanobacteria (Cyanobacteria). The presence of microphototrophs was detected throughout the year but showed spatio-temporal variations. Microphototrophs were absent in the parts of Veternica Cave with seeping and dripping water, while they were occasionally present in the hydrologically active parts of the cave. The presence of diatoms in the aphotic zone of Veternica Cave was related to hydrological conditions, and was not affected by the distance from the cave entrance. The presence of microphototrophs in caves of the Dinaric karst has been associated with caves subject to various types of flooding by endogenous and exogenous water. Despite the fact that microphototrophs are passively transported to the caves from the surface habitats, the presence of live individuals in the cave aphotic zone implies that they should not be neglected in discussions about cave food webs. Future research of microphototrophs should be focused on the species identification, their abundance, survival time, and detail description of conditions that determine their presence in caves.

Testate amoebae are phylogenetically a very diverse group of eukaryotic microorganisms. They can be found in marine and freshwater habitats and in soil. Some of these single-celled organisms inhabit both surface and cave habitats, but their diversity in caves has barely been explored. Recent studies in the Dinaric region imply that testate amoebae in caves show a high diversity. The aim of this study was to identify the alpha diversity of testate amoebae in the Lika region (Dinaric karst, Croatia) and to compare the habitats of different caves based on testate amoebae assemblages. In eight caves we found more than 40 testate amoebae taxa, including a new testate amoeba species, Psammonobiotus dinaricasp. nov. The greatest diversity of testate amoebae was found in Markov ponor (27 taxa). The Bray-Curtis Similarity Index showed that testate amoebae assemblages in caves inhabited by the endemic and endangered cave bivalve Congeria jalzici (Markov ponor, Dankov ponor and Dražice ponor) differ from caves not inhabited by this species. This differentiation is attributed to the impact of the sinking Lika river, which occasionally completely submerges these caves, creating specific habitats for eukaryotic microorganisms. This study contributes to our understanding of the diversity, biogeography and ecology of testate amoebae in caves, as well as providing further insight into the conditions that sustain populations of C. jalzici.

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