European Lakes Research Articles

Microplastics and anthropogenic fibre concentrations in lakes reflect surrounding land use.

Pollution from microplastics and anthropogenic fibres threatens lakes, but we know little about what factors predict its accumulation. Lakes may be especially contaminated because of long water retention times and proximity to pollution sources. Here, we surveyed anthropogenic microparticles, i.e., microplastics and anthropogenic fibres, in surface waters of 67 European lakes spanning 30° of latitude and large environmental gradients. By collating data from >2,100 published net tows, we found that microparticle concentrations in our field survey were higher than previously reported in lakes and comparable to rivers and oceans. We then related microparticle concentrations in our field survey to surrounding land use, water chemistry, and plastic emissions to sites estimated from local hydrology, population density, and waste production. Microparticle concentrations in European lakes quadrupled as both estimated mismanaged waste inputs and wastewater treatment loads increased in catchments. Concentrations decreased by 2 and 5 times over the range of surrounding forest cover and potential in-lake biodegradation, respectively. As anthropogenic debris continues to pollute the environment, our data will help contextualise future work, and our models can inform control and remediation efforts.

Return of the Apex Predator — How Brown Trout (Salmo trutta) Re-Establishment Shapes an Ecosystem

Re-establishment of a declined apex predator fish species in a lake ecosystem may have dramatic effects on other fish and plankton community already inhabiting the ecosystem. We studied mechanistically potential impacts of re-establishment of the brown trout (Salmo trutta) in the west-central European Lake Constance focusing on two commercially important fish species: whitefish and Eurasian perch. We compared simulation model outputs from two versions of an allometric trophic network model for Lake Constance, one with and one without the trout as the apex predator. The re-establishment of the declined brown trout reduced the perch population directly by predation and indirectly by increased resource competition, whereas the whitefish population was directly affected by increased predation. The decrease in fish biomass densities was highly sensitive to trout larval survivability. Both species showed strong compensatory behaviour by achieving higher per capita resource consumption when the populations decreased in the presence of brown trout.

A multi-omics study on quantifying antimicrobial resistance in European freshwater lakes.

The surveillance of wastewater for the Covid-19 virus during this unprecedented pandemic and mapped to the distribution and magnitude of the infected in the population near real-time exemplifies the importance of tracking rapidly changing trends of pathogens or public health problems at a large scale. The rising trends of antimicrobial resistance (AMR) with multidrug-resistant pathogens from the environmental water have similarly gained much attention in recent years. Wastewater-based epidemiology from water samples has shown that a wide range of AMR-related genes is frequently detected. Albeit sewage is treated before release and thus, the abundance of pathogens should be significantly reduced or even pathogen-free, several studies indicated the contrary. Pathogens are still measurable in the released water, ultimately entering freshwaters, such as rivers and lakes. Furthermore, socio-economic and environmental factors, such as chemical industries and animal farming nearby, impact the presence of AMR. Many bacterial species from the environment are intrinsically resistant and also contribute to the resistome of freshwater lakes. This study collected the most extensive standardized freshwater data set from hundreds of European lakes and conducted a comprehensive multi-omics analysis on antimicrobial resistance from these freshwater lakes. Our research shows that genes encoding for AMR against tetracyclines, cephalosporins, and quinolones were commonly identified, while for some, such as sulfonamides, resistance was less frequently present. We provide an estimation of the characteristic resistance of AMR in European lakes, which can be used as a comprehensive resistome dataset to facilitate and monitor temporal changes in the development of AMR in European freshwater lakes.

Minor Treatments Can Play a Significant Role in Preserving Natural Habitats and Protected Species on the Shore of a Central European Lake

Semi-natural grasslands were previously established through traditional land use and maintained by active management, but their extension nowadays is declining rapidly, particularly in areas that also have tourism potential. In parallel, the conservation value of the remaining areas is increasing. The shore of Lake Balaton is a particularly good example, as Lake Balaton is an area highly affected by tourism, yet there have been valuable habitats able to survive and provide refuge for many vulnerable, protected species. Fortunately, we have reliable information about the vegetation of the area from two decades ago. Comparing these data with our recent surveys we investigated the changes in habitats and the distribution of protected plant species in connection with the active conservation treatments such as grazing or cutting. Our results show that in areas where treatments are still ongoing, protected plant species are more likely to survive, or even other species can appear, which is in clear contrast with conditions experienced in abandoned areas, where at least seven protected species have disappeared. According to our results, minor, but appropriately chosen and well-executed management interventions, can help in the long-term maintenance of species-rich habitats and improving the conservation status of threatened species.

Inventory of the benthic eukaryotic diversity in the oldest European lake.

We have profound knowledge on biodiversity on Earth including plants and animals. In the recent decade, we have also increased our understanding on microorganisms in different hosts and the environment. However, biodiversity is not equally well studied among different biodiversity groups and Earth's systems with eukaryotes in freshwater sediments being among the least known. In this study, we used high‐throughput sequencing of the 18S rRNA gene to investigate the entire diversity of benthic eukaryotes in three distinct habitats (littoral sediment and hard substrate, profundal sediment) of Lake Ohrid, the oldest European lake. Eukaryotic sequences were dominated by annelid and arthropod animals (54% of all eukaryotic reads) and protists (Ochrophyta and Ciliophora; together 40% of all reads). Eukaryotic diversity was 15% higher in the deep profundal than on either near‐surface hard substrates or littoral sediments. The three habitats differed in their taxonomic and functional community composition. Specifically, heterotrophic organisms accounted for 92% of the reads in the profundal, whereas phototrophs accounted for 43% on the littoral hard substrate. The profundal community was the most homogeneous, and its network was the most complex, suggesting its highest stability among the sampled habitats.

A new species of Sclerocephalus with a fully ossified endocranium gives insight into braincase evolution in temnospondyls

AbstractThe late Paleozoic temnospondyl Sclerocephalus formed an aquatic top predator in various central European lakes of the late Carboniferous and early Permian. Despite hundreds of specimens spanning a wide range of sizes, knowledge of the endocranium (braincase and palatoquadrate) remained very insufficient in Sclerocephalus and other stereospondylomorphs because even large skulls had unossified endocrania. A new specimen from a stratigraphically ancient deposit at St. Wendel in southwestern Germany is recognized as representing a new taxon, S. concordiae new species, and reveals a completely ossified endocranium. The sphenethmoid was completely ossified from the basisphenoid to the anterior ethmoid region, co-ossified with the parasphenoid, and the basipterygoid joint was fully established. The pterygoid bears a slender, S-shaped epipterygoid, which formed a robust pillar lateral to the braincase. The massive stapes was firmly sutured to the parasphenoid. In the temnospondyl endocranium, character evolution involved various changes in the epipterygoid region, which evolved distinct morphologies in each of the major clades.UUID: http://zoobank.org/5e6d2078-eacf-4467-84cf-a12efcae7c0b

Land Use and Land Cover Pattern as a Measure of Tourism Impact on a Lakeshore Zone

Lakes provide different ecosystem services, including those related to tourism and recreation. Sustainable development principles should be respected in lake tourism planning. The aim of this study was to assess the impact of tourism on the lakeshore zone in a typical post-glacial Lakeland in Northern Poland (Central Europe). An explanatory analysis of the distribution of individual spatial factor values was performed using the SHapley Additive exPlanations algorithm (SHAP). In a first step, the aim was to select a Machine Learning model for modelling based on Shapley values. The greater or lesser influence of a given factor on the tourism function was measured for individual lakes. The final results of ensemble modelling and SHAP were obtained by averaging the results of five random repetitions of the execution of these models. The impact of tourism on the lakeshore zone can be much more accurately determined using an indirect method, by analysing the tourism and recreational infrastructure constantly present there. The values of the indices proposed in the study provide indirect information on the number of tourists using the tourist and recreational facilities and are a measure of the impact of tourism on the lakeshore zone. The developed methodology can be applied to the majority of post-glacial lakes in Europe and other regions of the world in order to monitor the threats resulting from shore zone exploitation. Such studies can be an appropriate tool for management and planning by the relevant authorities.

Assessment of the spread of pollution in the riverbed of the Neva River as a result of an emergency at a toxic waste landfill

The catchment area of the Neva River, which includes the largest European lakes Ladoga, Onega, Ilmen and Saimaa, located in Finland, has an area of 183,300 km2. The quality of water in the Neva River is determined by the characteristics of the water masses of Lake Ladoga, as well as by point and non-point sources of pollution on the tributaries of the Neva. One of the potential sources of pollution is the Krasny Bor landfill for storing highly toxic waste of hazard classes I – V. In the event of an emergency at the landfill, toxic wastewater can enter the Izhora River, and then into the Neva River, 7 km above the water intakes of St. Petersburg. Using the methods of mathematical modeling, the calculation of the spread of toxic substances in the hydrographic network of the catchment area, and then in the channel of the Neva River was carried out under various scenarios of the development of an emergency at the landfill. A full-scale experiment was carried out to confirm the adequacy of the model of impurity spreading in the channel of the Neva. An assessment of the features of the transport of suspended particles and dissolved impurities in the direction of the water intakes of St. Petersburg is given.

Assessing the ecological vulnerability of the shallow steppe Lake Neusiedl (Austria-Hungary) to climate-driven hydrological changes using a palaeolimnological approach

Lake Neusiedl, the largest steppe lake in Europe, is particularly sensitive to climate variations due to its extreme shallowness (zmax = 1.8 m) and low ratio of catchment to lake area (3.5 : 1). Changes in water budget, salinity and turbidity have key implications for the lake’s ecology and management. Here, we present a multi-proxy palaeolimnological reconstruction of the evolution of Lake Neusiedl since the end of its last complete desiccation (1865–1868), based on an undisturbed radiometrically dated core taken from the open water portion of the lake. Geochemical and biological (algal) proxies outline the succession of three major ecological stages since 1873 ± 16 yrs, with the first major changes appearing already in the 1930s as driven by climate related hydrological variability. Subfossil diatoms proved to be reliable for tracking long-term changes in the trophic conditions of Lake Neusiedl while diatom-inferred lake conductivity revealed to be unreliable due to a combination of lake environmental settings and the absence of a site-specific training set. Nonetheless, multivariate statistical analyses and comparisons with limnological data confirm a great potential of subfossil diatoms for revealing past ecological changes and tipping points of shallow lakes, as long as studies rely on a multi-proxy approach. In agreement with limnological surveys, the sediment record corroborates the high vulnerability of Lake Neusiedl, both in present and past times, towards climate-driven changes in water level and salinity, and allows the prediction, by analogy with the past, of future ecological changes in a context of global warming and increasing nutrient inputs from non-point sources.

Habitat partitioning among three predatory fish in a temperate reservoir

AbstractThe diversity of predatory species plays a key role in ecosystem functioning but our understanding of the mechanisms underlying their coexistence is limited, particularly in freshwater ecosystems. Northern pike Esox lucius, European perch Perca fluviatilis and pikeperch Sander lucioperca are three widespread predatory species in European lakes, where they often coexist. As potential competitors, we hypothesised that partitioning habitat is a determinant of species coexistence. This was tested by quantifying the variability of their habitat use in tracking adult individuals in the Bariousses reservoir (France, 86.6 ha, mean depth 7.1 m). Specifically, we investigated their distribution along the littoral–pelagic and depth axes along the daily cycle and across seasons. From littoral to pelagic waters were first found pike, then perch and finally pikeperch. Pike was the closest to the surface while pikeperch was the deepest. This general pattern was, however, variable across seasons with the three species located in the upper layer in summer during reservoir stratification. Individuals were more evenly distributed along the littoral–pelagic axis and closer to the bottom when water was mixing (autumn, winter). In summer, perch used more intensively in the pelagic zone during the daytime. Other species did not show any diel change of habitats. Our results highlighted that species coexistence is associated with habitat partitioning among these three predators, with perch showing a more variable behaviour regarding habitat characteristics. Now more than ever, in the context of global change which modifies habitats, it is of crucial importance to understand the coexistence mechanisms of species that shape ecosystems.

Diversity of Algae and Cyanobacteria and Bioindication Characteristics of the Alpine Lake Nesamovyte (Eastern Carpathians, Ukraine) from 100 Years Ago to the Present

The species diversity and changes in the structural dynamics of the algal flora from the alpine lake Nesamovyte has been studied for 100 years. During the period of investigations, 234 species (245 infraspecific taxa) were revealed to cover more than 70% of the modern species composition of the studied lake. The modern biodiversity of algae is characterized by an increase in the number of widespread forms, a change from the baseline “montane” complex in comparison to the beginning of the 20th century. Nevertheless, the Nesamovyte Lake still has a unique algae composition that is typical for high-mountainous European lakes. The presence of a different complex of conventionally arctic species of algae, in particular, diatoms is discussed. Structural changes in the taxonomic composition of the algal flora of the lake as well as in the complex of the leading genera, species and their diversity are revealed. An ecological analysis of the algal species composition of the lake showed vulnerability and degradation to the ecosystem of the lake. On this basis, the issue regarding the question of protection and preservation of the algae significance and uniqueness of the flora of algae in the Nesamovyte Lake are discussed.

Cyanophage Distribution Across European Lakes of the Temperate-Humid Continental Climate Zone Assessed Using PCR-Based Genetic Markers

Studies of the diversity and distribution of freshwater cyanophages are generally limited to the small geographical areas, in many cases including only one or few lakes. Data from dozens of various lakes distributed at a larger distance are necessary to understand their spatial distribution and sensitivity to biotic and abiotic factors. Thus, the objective of this study was to analyze the diversity and distribution of cyanophages within the infected cells using marker genes (psbA, nblA, and g91) in 21 Polish and Lithuanian lakes. Physicochemical factors that might be related to them were also analyzed. The results demonstrated that genetic markers representing cyanophages were observed in most lakes studied. The frequently detected gene was psbA with 88% of cyanophage-positive samples, while nblA and g91 were found in approximately 50% of lakes. The DNA sequence analyses for each gene demonstrated low variability between them, although the psbA sequences branched within the larger cluster of marine Synechoccocuss counterparts. The principal component analysis allowed to identify significant variation between the lakes that presented high and low cyanobacterial biomass. The lakes with high cyanobacterial biomass were further separated by country and the different diversity of cyanobacteria species, particularly Planktothrix agardhii, was dominant in the Polish lakes and Planktolyngbya limnetica in the Lithuanian lakes. The total phosphorous and the presence of cyanophage genes psbA and nblA were the most important factors that allowed differentiation for the Polish lakes, while the pH and the genes g91 and nblA for the Lithuanian lakes.

Model‐based decomposition of environmental, spatial and species‐interaction effects on the community structure of common fish species in 772 European lakes

AbstractAimWe tested whether there is a strong effect of species interactions on assembly of local lake fish communities, in addition to environmental filters and dispersal.LocationSeven hundred and seventy‐two European lakes and reservoirs.Time period1993–2012.Major taxa studiedNineteen species of freshwater fishes.MethodsWe applied a latent variable approach using Bayesian Markov chain Monte Carlo algorithms (R package “BORAL”). We compared the contributions of six environmental predictors and the spatial organization of 772 European lakes in 209 river basins on the presence/absence of the 19 most frequent fish species and on the biomass and mean mass of the six dominant species. We inspected the residual correlation matrix for positive and negative correlations between species.ResultsEnvironmental (50%) and spatial (10%) predictors contributed to the presence/absence assembly of lake fish communities, whereas lake size and productivity contributed strongly to the biomass and mean mass structures. We found highly significant negative correlations between predator and prey fish species pairs in the presence/absence, biomass and mean mass datasets. There were more significantly positive than negative correlations between species pairs in all three datasets. In addition, unmeasured abiotic predictors might explain some of the correlations between species.Main conclusionsStrong effects of species interactions on assembly of lake fish communities are very likely. We admit that our approach is of a correlational nature and does not generate mechanistic evidence that interactions strongly shape fish community structures; however, the results fit with present knowledge about the interactions between the most frequent fish species in European lakes and they support the assumption that, in particular, the mean masses of fish species in lakes are modified by species interactions.

Atmospheric deposition of semivolatile organic pollutants in European high mountains: Sources, settling and chemical degradation

Bulk atmospheric deposition samples, including wet and dry deposition, were collected during 2004–2006 in four high mountain European lakes: Skalnate Pleso (Tatra mountains, Slovakia), Gossenköllesee (Alps, Austria), Redon (Pyrenees, Spain), and Lochnagar (Grampian Mountains, Scotland). Samples were analysed for polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCHs), endosulfans, and polybromodiphenyl ethers (PBDEs).The deposition of PCBs, HCHs, and low brominated BDEs reflected baseline contributions from long range atmospheric transport. This was also the case for PAHs in Redon and Gossenköllesee, endosulfans in Lochnagar and Gossenköllesee and HCB in these three lakes. However, Skalnate received PAHs, endosulfans, and HCB from regional sources as it was the case for endosulfans in Redon. The distinct origin of these pollutants was reflected in the relative composition of some metabolites such as the proportion of endosulfan sulfate vs α- and β-endosulfans or the relative composition of BDE47 and BDE99.Wet deposition was the main process for atmospheric removal of PAHs, HCHs, and HCB. In addition, warm season revolatilization from soils and melting snow with subsequent condensation at low temperature were significant for volatile PAHs, HCB, low chlorinated PCBs, and endosulfans.Reaction with OH radicals was not a significant loss process of HCHs and HCB in remote areas, dominated by wet deposition, whereas PCBs and PAHs were significantly removed by both wet deposition and OH radical oxidation, the latter dominating in the highest altitude sites. Photolysis was the main atmospheric removal process of PBDEs, dominating over atmospheric deposition and OH depletion in all sites.

<strong><em>Pseudostaurosira crateri sp. nov.</em> (Fragilariaceae, Bacillariophyta), a new small araphid, fossil diatom species from the Pleistocene (Atlantic Forest, Brazil)</strong>

Pseudostaurosira crateri sp. nov. is described from fossil material analyzed from a Quaternary sediment core retrieved from the Colônia basin (located in the Brazilian Coastal Plain, Atlantic Forest, São Paulo Metropolitan Region). Light and scanning electron microscopy revealed its morphology and enabled its comparison with morphologically similar species from European and African rivers and lakes. The new species features wide and short vimines, a particular characteristic of the genus Pseudostaurosira, in addition to many other exclusive features that distinguish it from any other species in the genus reported to date. Some of these characteristics are: (i) apical pore fields composed of small poroids placed within cavernous troughs carved deep into the valve, (ii) absence of blisters or depositions along the valve mantle edge, (iii) small-sized valve, (iv) presence of incipient or rudimentary spines located on raised virgae at the valve face/mantle junction, and (v) internal accumulations of siliceous material on the volae appearing as concentric discs partially occluding the depression into which the areolae open internally. We further discuss the morphological features of similar species ascribed to the genera Pseudostaurosira, Pseudostaurosiropsis, Staurosirella, Frankophila, and Popovskayella based on literature information. Finally, this study contributes to the knowledge on diatom diversity and its biogeography, especially those from tropical regions, in addition to providing a taxonomical basis for its use as biological markers for paleoenvironmental reconstruction.

Increasing maximum lake surface temperature under climate change

Annual maximum lake surface temperature influences ecosystem structure and function and, in particular, the rates of metabolic activities, species survival and biogeography. Here, we evaluated 50 years of observational data, from 1966 to 2015, for ten European lakes to quantify changes in the annual maximum surface temperature and the duration above a potentially critical temperature of 20 °C. Our results show that annual maximum lake surface temperature has increased at an average rate of +0.58 °C decade−1 (95% confidence interval 0.18), which is similar to the observed increase in annual maximum air temperature of +0.42 °C decade−1 (95% confidence interval 0.28) over the same period. Increments in lake maximum temperature among the ten lakes range from +0.1 in the west to +1.9 °C decade−1 in the east. Absolute maximum lake surface water temperatures were reached in Wörthersee, 27.5 °C, and Neusiedler See, 31.7 °C. Periods exceeding a critical temperature of 20 °C each year became two to six times longer than the respective average (6 to 93). The depth at which water temperature exceeded 20 °C increased from less than 1 to more than 6 m in Mondsee, Austria, over the 50 years studied. As a consequence, the habitable environment became increasingly restricted for many organisms that are adapted to historic conditions.

Quantification of the covariation of lake microbiomes and environmental variables using a machine learning-based framework.

It is known that microorganisms are essential for the functioning of ecosystems, but the extent to which microorganisms respond to different environmental variables in their natural habitats is not clear. In the current study, we present a methodological framework to quantify the covariation of the microbial community of a habitat and environmental variables of this habitat. It is built on theoretical considerations of systems ecology, makes use of state-of-the-art machine learning techniques and can be used to identify bioindicators. We apply the framework to a data set containing operational taxonomic units (OTUs) as well as more than twenty physicochemical and geographic variables measured in a large-scale survey of European lakes. While a large part of variation (up to 61%) in many environmental variables can be explained by microbial community composition, some variables do not show significant covariation with the microbial lake community. Moreover, we have identified OTUs that act as "multitask" bioindicators, i.e., that are indicative for multiple environmental variables, and thus could be candidates for lake water monitoring schemes. Our results represent, for the first time, a quantification of the covariation of the lake microbiome and a wide array of environmental variables for lake ecosystems. Building on the results and methodology presented here, it will be possible to identify microbial taxa and processes that are essential for functioning and stability of lake ecosystems.

Unravelling the effect of multiple stressors on ecological structure of littoral lake macroinvertebrates

AbstractMorphological alteration of shorelines and eutrophication both affect the biological integrity of European lakes. These pressures, often acting simultaneously, are difficult to tease apart. In this study, we related the number of taxa with specific habitat preference to habitat complexity across lakes of varying nutrient state. Habitat complexity at morphologically altered shorelines was significantly lower than at unaltered sites across trophic categories. A generalised linear mixed‐effects model showed decreased number of taxa with specific mesohabitat preference at morphologically simplified sites in oligotrophic and mesotrophic, but not eutrophic lakes. These results suggest: (1) an antagonistic interaction between the effect of nutrient enrichment and morphological alterations on lake littoral communities and (2) the number of macroinvertebrate habitat specialists could potentially be used to assess the effects of structural simplifications of shorelines in lakes of low to medium nutrient status. We conclude that the use of functional traits approach in aquatic ecology should foster better understanding of stressor–response relationships for combined effect of multiple stressors.

First insights into the diversity and ecology of non-biting midges (Diptera: Chironomidae) of the unique ancient Skadar Lake basin (Montenegro/Albania)

In the present study, based on faunistic information collected during 2014 and 2015, in association with physical–chemical conditions of the collection sites, the environmental factors influencing the composition and distribution of chironomid assemblages were investigated. In total, 164 Chironomidae taxa reported in this study extended the existing checklist with 152 taxa newly found in the Skadar Lake basin. The comparison based on the list of species from the six large, well-studied European lakes showed that Lake Constance (Switzerland/Germany/Austria) is the most species-rich waterbody with 174 taxa, followed by Lake Skadar with 164 taxa. For pupal exuviae, the most species-rich site was situated in the centre of the small lake (north-western part of the Skadar Lake). A Canonical Correspondence Analysis (CCA) revealed that shallow coastal parts of the lake are much richer in species than the open, deeper parts of the lake. This first investigation since 1979 of non-biting midge species (including imagines) diversity will help to monitor biodiversity of the basin and to understand how protection programs influence biodiversity within the lake basin.

DNA barcoding in recognition of Gammarusflock diversity and distribution in the ancient Lake Ohrid

Lake Ohrid, located on the Balkan Peninsula at the Albanian-Macedonian border, is the oldest European lake (1.3-1.9 My old) and one of the world’s smallest ancient lakes. Taking into account the size of the lake and its biodiversity, it harbors the highest level of endemism, especially within amphipod crustaceans (ca. 90%) with the endemic Gammarus species flock. Our previous studies upon this flock have shown a substantial decoupling between molecular and morphological diversity, existence of cryptic species and puzzling speciation history. In order to explore sources of observed diversity, in the current study we are investigating ecological preferences of the species within the flock, based on their distribution in depth gradient, in relation to molecular diversity based on DNA barcoding. In the study over 200 barcodes were generated and combined with 173 previously published. The specimens were collected from all depth ranges of Lake Ohrid as well as from springs located on or near the banks of the lake. Within the species flock, 13 BIN’s were identified, 12 previously known and one newly recognized, representing separate lineage and putatively a new species. Two of the flock species were found only in the springs: G. sketi and G. cryptosalemaai. G. sketi, previously found only in springs on the southern banks of Lake Ohrid, has now also been discovered in springs in its north-eastern part. Both species show low haplotypic diversity. All remaining species were recorded from the depth between 20 and 60 meters, that is characterized by the highest ecological diversity with different types of substrates: stones, macrophytes, abundant Dreissena shells as well as sand and silt. Among them G. sywulai, G. macedonicus, G. cryptoparechiniformis, G. lychnidensis, G. ochridensis, G. parechinifromis were found exclusively within this depth range. The three latter species represent single BIN and share haplotypes, at the same time this BIN has the highest number of haplotypes in comparison to others. The remaining species found on this depth represent separate BINs with different levels of haplotype diversity. Only G. lychnidensis, G. stankokaramani and G. solidus were found below the depth of 60 meters, in a quite homogenous environment dominated by silt. In the deepest parts of the lake, between 260 and 290 meters, only G. solidus was found. This species is represented only by three haplotypes while G. stankokaramani is characterized by multiple haplotypes partially shared with G. lychnidensis. The shared haplotype represents the only G. lychnidensis occurrences on the depths below 60 meters. Summarizing, the highest abundance of BINs, species and haplotypes was recovered from the most ecologically diversified depth range of the lake (20 to 60 meters). This suggests that ecological heterogeneity could be the main driver of Gammarus species flock diversification in the ancient Lake Ohrid. Due to the complex pattern of morphological diversity, DNA barcoding proved to be the best if not the only method in identification of the species flock diversity.