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Mires are specific freshwater habitats with great importance for global biodiversity. The present study investigated diatoms in different habitats of a Đon Močvar mire and their relationship with environmental parameters, as well as the use of diatom indicator values and the German Red List for conservation purposes. The study was conducted monthly from January to November 2012. Diatoms were sampled simultaneously with water for physical and chemical properties in three different habitats. A total of 50 diatom taxa were identified. Based on their relative abundance and diversity, habitat-related environmental factors shaped the diatom community when seasonal dynamics were less important. Multivariate analysis showed that chemical oxygen demand (COD) and conductivity were the most significant environmental variables describing diatom communities. The indicator values characterized the diatoms as a typical mire community and proved to be a reliable starting point for future monitoring and management. Diatom classification by German Red List revealed a dominance of species with priority for conservation, suggesting that the Đon Močvar should have conservation status. This study brings a better understanding of the response of diatoms to living in specific habitats and the basis for future studies on ecological characteristics of new or rare taxa.

Phytoplankton represents one of the most important biological components of primary production, trophic interactions and circulation of organic matter in lakes and reservoirs. To contribute to the understanding of eutrophication processes and ecological status of the small, shallow Butoniga reservoir, a machine learning tool for induction of models in form of decision trees and rule-based models was applied on a data set comprising physical, chemical and biological variables measured at four stations. Two types of models were successfully elaborated, i.e. (1) model describing phytoplankton Phylum which describes and connects phytoplankton Phylum with phytoplankton abundance and biomass, and (2) model simulating phytoplankton biomass according to environmental variables which could be used in management purposes. Such models and their presentation contribute to a better understanding of the Butoniga reservoir ecosystem functioning.

Ciliates are a group of phagotrophic protists found in a wide variety of ecosystems. This study builds on recent studies of ciliates in the Krka river and investigates changes in the phylogenetic and functional diversity of ciliates in biofilm to predict the phylogenetic and functional structure of ciliates in other karstic rivers. Biofilm samples were collected from four representative locations: upstream (Krka spring), midstream (Marasovine), and downstream (Roški slap, Skradinski buk) of the Krka river to test for differences in phylogenetic and functional diversity of ciliates in relation to location and positioning on tufa stones (light/dark-exposed side of tufa stone). Our results showed that Krka spring had higher phylogenetic species variability, lower phylogenetic diversity, and lower functional richness than Skradinski buk, suggesting phylogenetic overdispersal at Krka spring. This could be due to environmental filtering, competitive exclusion, or a combination of these factors. As the first study of its kind in the Mediterranean, our results shed light on the phylogenetic and functional diversity of ciliates in karst ecosystems and provide a basis for future ecological and conservation efforts.

A new marine strain of picoplanktonic algae, PMPFPPE4, was isolated from a mixed net-phytoplankton sample taken from the upper euphotic layer of the southeastern Adriatic Sea. Evaluation of the new strain included morphological investigation (by light and electron microscopy), phylogenetic analysis (utilizing plastid 16S rRNA and nuclear 18S rRNA genes), and physiological characterization (screening of pigment/lipid composition and capturing photosynthesis measurements). The new strain was proven to belong to the genusPicochlorumand the lipid composition revealed an unexpected accumulation of triacylglycerols, indicating an evolutionary adaptation for growth under unfavourable conditions. In addition, lipid remodelling in the exponential to stationary growth phase was characterized by an increased share of membrane-forming digalactosyldiacylglycerols and phosphatidylcholines. Maximum photosynthetic activity measured was at 30 degrees C, but the most rapid increase of photosynthetic activity was at lower temperatures (15-20 degrees C). Moreover, the thermotolerant strain did not exhibit photoinhibition below 40 degrees C and survived a one-month cultivation period in complete darkness. The strain's survival in low light and dark conditions suggests a potential shift from autotrophy to mixotrophy under unfavourable growth conditions. Thus, the unique physiological attributes represented by a high growth rate, thermotolerance, phototolerance and high triacylglycerol synthesis may render the strain highly attractive for biofuel production and growth in large outdoor systems.

Due to its geographical position, relief features, specific geological past and great heterogeneity of space in a relatively small area, Bosnia and Herzegovina has specific hydro-morphological features, with numerous spring habitats and unique aquatic biodiversity, mainly composed by stenothermic and relict taxa. Spring habitats, as well as the flora and fauna that inhabit them are considered particularly vulnerable to climate change and anthropogenic pressures, due to their isolation and fragmentation. According to forecasts, these ecosystems will be exposed to multiple pressures caused by changes in water regime, rising water temperatures and a continuous decline in water quality, which will lead to the disappearance of more sensitive crenobiont species and a reduction in biodiversity. Therefore, in this chapter we present the ecological characteristics and specifics of spring habitats in Bosnia and Herzegovina, with a preliminary list of species that inhabit them and special focus on endemic, sensitive and rare species, which are potentially threatened by predicted changes in habitat quality.

Karst ecosystems play a unique role as exceptional natural habitats in sustaining biodiversity. This study focuses on diatoms, a diverse group of microeukaryotes in the periphytic community of a karstic river. In a multi-microhabitat study along the Krka River (Croatia), our goal was to obtain a detailed overview of diatom diversity and community structure using morphological and molecular approaches, and to assess the applicability of eDNA metabarcoding as a reliable tool for biomonitoring assessment. The results revealed a relatively low agreement in the diatom community composition between the two approaches, but also provided complementary information, with no differences in beta diversity detected between microhabitats. The SIMPER analysis underlined the importance of the molecular approach in identifying diatom community composition, due to errors in distinguishing between deposited diatom cells that occurred in the morphological analysis. In contrast, the morphological approach indicated a clear diatom community separation along the river with a strong location effect. Despite certain differences, both approaches provided a feasible assessment of the ecological status according to the relationship to environmental pressures, classifying the Krka River as High (morphological approach) or Good (molecular approach) throughout the most of its course. Moreover, diatom diversity based on both approaches provides a reliable dataset applicable in routine monitoring assessment and offers a deeper understanding of the presented ecological status. The incompleteness of a reference database presents one major drawback of the molecular approach, which needs further updating in order to improve routine diatom metabarcoding.

The biodiversity of algal communities and environmental conditions were investigated in the springs of Mt. Konjuh. The assemblages of 20 springs emerging from different lithologies (limestones and ophiolites, respectively) comprised 234 algal taxa. Diatoms and cyanobacteria were the most species-rich groups. The most common alkaliphilic, circumneutral, and eutraphentic diatoms were represented by the genera Gomphonema, Nitzschia, Navicula, Cymbella, and Achnanthidium, and by the cyanobacterial genus Phormidium. Hierarchical clustering and SIMPROF analysis based on relative algal abundance clustered springs into six groups, separating them mainly according to spring type and lithology. Indicator species for groups and springs on different lithological substrata were singled out, revealing 33 taxa with preferences for ophiolites, and 20 taxa with preferences for carbonates. The values of the Shannon-Wiener diversity index were moderately high per spring location, and similar for the two groups of springs on different lithologies. A higher similarity in species composition was noted between springs on ophiolites and limestones than between springs on ophiolites and other types of siliceous substrata. The present study suggests that algal assemblages in springs emerging from ophiolites, even those made up by a preponderance of silicates, should be analyzed separately from those related to springs on other siliceous substrata. The results obtained showed that most of the springs studied are affected by anthropogenic impacts and morphological alterations leading to the dominance of highly competitive meso- and eutraphentic algal species, thus emphasizing the importance of further investigation and conservation of these habitats.

EnglishThe spatial and temporal patterns of phytoplankton in relation to physical and chemical properties were studied in a drinking water reservoir - the Lazberc Reservoir - located in north-east Hungary. The main objectives were to determine the role of the hydrologic regime in structuring algal growth and the effects of physical and chemical variables on the coexistence of a dominant species: cyanobacteria, Microcystis aeruginosa (Kutzing) Kutzing and an eukaryotic dinoflagellates, Ceratium hirundinella (O.F. Muller) Dujardin. The results suggest that nutrients (TP, NH4–N and NO3–N), turbidity, and the hydrologic regime play an important role in regulating the occurrence of investigated species. The rainy summer period resulting in a mixing of the water column, and the low level of nitrogen coupled with the internal release of phosphorus from the lake sediment under brief periods of anoxia promote cyanobacterial bloom. During this period, the water column was characterised by high turbidity, which created favourable conditions for developing a huge C. hirundinella biomass and the simultaneous occurrence of M. aeruginosa. This study also demonstrated - that contrary to what was previously known, deep standing waters in the temperate zone are not only dimictic (mixing twice a year during the spring and the fall), but they can also be mixed during the summer period due to continued rainfall, which equalizes the temperature in the whole water column. This can promote the occurrence of cyanobacterial and eukaryote planktonic organisms portuguesOs padroes espaciais e temporais do fitoplâncton e sua correlacao com as especificidades variaveis fisicas e quimicas, foram examinados no nordeste da Hungria, localizados em um reservatorio tipico de agua potavel no reservatorio de Lazberc. Nosso principal objetivo foi para esclarecer o papel da hidrologia, e os diferentes especificidades fisicas e quimicas na coexistencia de especies dominantes: cianobacteria, Microcystis aeruginosa (Kutzing) Kutzing e Dinoflagellata eucariotica, Ceratium hirudinella (OF Muller) Dujardin. De acordo com os resultados, os nutrientes (TP, NH4-N, NO3-N), turbidez e os condicoes hidrologicas desempenham um papel importante na regulacao da ocorrencia das especies examinadas. O periodo de verao chuvoso que resulta um equilibrio do nivel da agua, o baixo nivel de nitrogenio, juntamente com a liberacao interna de fosforo do sedimento, durante um curto periodo anoxico promove a floracao de cianobacterias. Durante este periodo, a coluna de agua foi caracterizada por alta turbidez, criando condicoes favoraveis para o surgimento de uma enorme expansao da biomassa C. hirundinella e a aparicao simultânea de M. aeruginosa. Este estudo mostrou tambem que as aguas profundas nas areas da zona moderada, nao tem apenas dimictic especialidade (misturando duas vezes por ano, na primavera e no outono), mas tambem durante a estacao do verao pode ser misturada por chuvas continuas que compensam a temperatura em toda a coluna da agua. Isso ajuda a presenca de cianobacterias e organismos eucariotas planctonicos

Ciliates (protozoa) are a very large and diverse group of microeukaryotes that occupy a central position in the trophic web of freshwater ecosystems. Ciliates exhibit high ecological sensitivity and have tremendous bioindicator potential, but they are largely ignored in routine biomonitoring, mainly due to limitations in morphological diagnosis, which is both time-consuming and costly. In this study, we combined molecular (Illumina sequencing of the hypervariable V9 region of the SSU rRNA gene) and morphological approaches to get a detailed insight into the periphytic ciliate community structure and function within a karst, tufa-precipitating hydrosystem (National Park Krka, Croatia). Periphyton (biofilm) was sampled from light- and dark-exposed lithified tufa/stones at four representative locations within the Krka River, including upstream, midstream, and downstream river sections. We identified hydrological parameters and saprobiological classification of the sampling locations as the main structuring factors for ciliate communities. The molecular approach showed a clear separation of sampling locations in beta diversity analyses, as confirmed by PERMANOVA test. In contrast, alpha diversity was mostly affected by light exposition of the lithified tufa/stones, as confirmed by morphological approach. Environmental conditions consistently affected ciliate community composition at all locations, resulting in clear separation of periphyton samples. Our results indicate that ciliates should be considered as important bioindicators in monitoring tufa-forming rivers and streams, and the molecular approach (V9 region) proved to be applicable because as it efficiently captured the diversity of ciliates in periphyton. Validation of such an approach would lead to significant progress in extending monitoring to a broader range of indicator organisms than those now included in standard monitoring.

The diatom genus Entomoneis is known from the benthos and plankton of marine, brackish, and freshwaters. Entomoneis includes diatoms with a bilobate keel elevated above the valve surface, a sigmoid canal raphe, and numerous girdle bands. Owing mostly to the scarcity of molecular data for a diverse set of species, the phylogeny of Entomoneis has not been investigated in depth. The few previous studies that included Entomoneis were focused on broader questions and the available data were from a small number of either unidentified Entomoneis or well-known species (e.g., E. paludosa). Since the first description of new species combining both molecular and morphological characters (E. tenera), we have continued to cultivate and investigate Entomoneis in the plankton of the Adriatic Sea. Combined multigene phylogeny (SSU rDNA sequences, rbcL, and psbC genes) and morphological observations (LM, SEM and TEM) revealed six new Entomoneis species supported by phylogenetic and morphological data: E. pusilla, E. gracilis, E. vilicicii, E. infula, E. adriatica, and E. umbratica. The most important morphological features for species delineation were cell shape, the degree and mode of torsion, valve apices, the appearance and structure of the transition between keel and valve body, the ultrastructure and the shape of the girdle bands, and the arrangement and density of perforations along the valve and valvocopulae. Our results highlight the underappreciated diversity of Entomoneis and call for a more in-depth morphological and molecular investigation of this genus especially in planktonic habitats.

An investigation into bryophyte communities in karst Jankovac Stream (Papuk Nature Park, Croatia) was carried out once a month from July 2008 to June 2009. Samples were taken from two lotic microhabitats: (i) Jankovac Spring (JS), a hypocrenal habitat with dense bryophyte clusters (90% bottom cover) and (ii) Jankovac Waterfall (JW), with scattered bryophyte clusters (50% bottom cover). At the same time, seston samples were collected during the spring as the source site and after the JW, as the outflow site. The goals of this study were to understand the (i) algal, protozoan and metazoan bryophyte community assemblages in these two lotic microhabitats, (ii) influence of environmental factors on the structuring of the bryophyte community and (iii) structure of seston along the longitudinal profile. A total of 172 taxa were determined: 68 algae, 55 protozoa, 24 meiofauna and 25 macroinvertebrates. Statistically significant differences between two microhabitats differing in percentage of bryophyte cover were established testing 13 environmental parameters. In dense bryophyte clusters, community structure was determined by flow velocity and pH, and macroinvertebrates achieved higher diversity and abundance. On the contrary, in scattered bryophyte coverage algae, protozoa and meiofauna reached higher abundance and diversity governed by the amount of suspended organic matter and epiphyton. In contrast to previous studies, the inverse ratio of community diversity and abundanceversuspercentage of bryophyte cover was established. We assume this to be the result of an enrichment of the scattered bryophyte clusters by upstream seston. Additionally, the effects of anthropogenic hydromorphological disturbance are reflected in macroinvertebrate diversity and abundance reduction.

Crveno jezero (Red Lake) is one of the largest karstic sinkhole lakes in the Europe. Diatom flora of this lake was unknown due to difficulties in collecting samples. Recently, samples were collected by speleodivers in the frame of systematical natural sciences research, during the cave diving expedition “Crveno jezero 2017” and observations revealed the existence of a new species, Gomphosphenia plenkoviciae Gligora Udovič & Žutinić sp. nov. The species is described herein as new to science based on light and scanning electron microscopy. Gomphosphenia plenkoviciae is characterized by having narrowly clavate to linear valves, which are slightly heteropolar in larger specimens to clavate in smaller specimens, thus differing from the closest taxon by the valve outline and shape of the central area. Smaller valves of G. plenkoviciae differ from other small-sized Gomphosphenia species by valve outline, striae morphology and the shape of axial and central areas.