Stakeholders Perceptions of Non-indigenous Freshwater Fish Species: A Case Study from a Mediterranean Biodiversity Hotspot.

Even though the ecological impacts of invasive alien species (IAS) have been well studied, social aspects of IAS in freshwater ecosystems are still poorly explored. This study aimed to assess the perceptions associated with non-indigenous freshwater fish species (NIFS) among environmental-related professional and non-professional stakeholders in a Mediterranean country that displays high levels of fish species endemism, by using a questionnaire survey approach. Overall, 203 individuals participated, from which, the majority (n = 144) were related to environmental sciences. Most of the respondents perceived NIFS to be a threat of national importance, possibly attributed to the emotional effect of the term non-indigenous (conceived as intruder). However, NIFS were indicated as the least important threat affecting aquatic ecosystems when compared with other pressures. This contrasting perception could be explained by the fact that broad-scale impacts (i.e., climate change) exert stronger reactions compared to NIFS. Interestingly, non-professionals were more successful in identifying NIFS and native species compared to professionals, however both indicated low overall identification success. The majority of the respondents, both professionals and non-professionals, considered that: (a) the official authorities do not implement actions for NIFS management, (b) government spending should be increased to manage NIFS even if it should be reduced for other needs, and (c) early detection is the best way to manage NIFS. The establishment of a nationwide network consisting of all related stakeholders on NIFS issues, aiming to public awareness and preventive management actions to limit the spread and impacts of NIFS should be set as a priority.

Evolutionary history of the Pelasgus minnows (Teleostei: Leuciscidae), an ancient endemic genus from the Balkan Peninsula.

The Balkans are one of the European biodiversity hotspots, hosting outstandingly rich, yet threatened, flora and fauna. This region hosts one of the richest endemic freshwater ichthyofauna in Europe, including several genera occurring exclusively here. One of these is the genus of the primary freshwater minnows Pelasgus, which was designated only in 2007. The genus is one of the most ancient genera of the family Leuciscidae and comprises seven small-bodied species, inhabiting slower, well-vegetated courses of rivers. This work is the first molecular multilocus phylogeny of the genus, based on one mitochondrial and three nuclear markers. In total, 305 individuals across almost entire distribution range of the genus were analysed. We inferred the evolutionary history of the species by comparing the results of our calibrated multilocus coalescent species-tree to palaeogeological events. The diversification of the genus started in the early Miocene and continued through to the beginning of Pleistocene. We identified the regions of the oldest colonization by Pelasgus, the drainages of the ancient lakes Ohrid and Prespa, and the southernmost part of the Peloponnese, hosting Pelasgus minutus, P. prespensis and P. laconicus, respectively. We showed that P. prespensis is not endemic to Lake Prespa, as previously thought; it occurs also in the Albanian River Devoll. This corroborates the emerging opinion that the endemic taxa of ancient lakes evolved within larger-scale historic drainages and not only within the lakes. Our results showed that the species with the most recent common ancestor of the early Pliocene origin, P. thesproticus, P. epiroticus, P. stymphalicus and P. marathonicus, have neighbouring distribution ranges. Pelasgus epiroticus is especially interesting, not only for its pronounced genetic diversity with a geographic pattern, but also for being found at three localities within the native distribution range of P. stymphalicus as a result of a translocation. At two of these localities, we identified hybrids between the two species, and at one of them, the genetically pure native species was not found at all. This points to a threat of the loss of the native ichthyofauna due to unintentional translocations.

Defining non-indigenous fish assemblage types in Mediterranean rivers: Network analysis and management implications.

The design and implementation of appropriate management actions to tackle the spread and negative impacts of non-indigenous fish species (NIFS) in freshwater ecosystems still remains a complex task. In an effort to address the limitations of current managerial approaches, our study develops and applies a classification framework to define non-indigenous fish assemblage types (FATs) in Mediterranean riverine ecosystems and identifies the linkage with various regional, local, biotic and abiotic environmental factors. This framework contributes to a pre-invasion stage screening and to the design of effective type-specific post-invasion management actions dealing with specific NIFS assemblages. A network analysis was applied in order to extract NIFS community level information from 393 samples obtained from 51 river basins of Greece, while a multivariate ordination analysis was conducted to detect the factors best explaining the structure and distribution of FATs. The association patterns of the study's inputs and outcomes are illustrated through an alluvial diagram, providing insights across different spatial scales. In total, five FATs were generated where the major NIFS contributors of average similarity within each modularity class revealed the key indicator species (Gambusia-FAT; Carassius/Lepomis-FAT; Pseudorasbora-FAT; Salmonids-FAT and Carp-FAT). Overall, the identified FATs varied spatially, indicating different community structure, mainly based on the diverse habitat preferences and life-history traits of indicator species. Alien FATs were mainly distributed within large and transboundary rivers, while Translocated and Salmonids FATs mostly occupied ecoregions with relatively depauperate faunas and often in biodiversity hotspots. The results of this study can identify conservation priorities within FATs, inform specific-type post-invasion management actions tackling NIFS, while in addition may provide valuable information for protecting high-priority water bodies before invasion.

Diversification in gravel beaches: A radiation of interstitial clingfish (Gouania, Gobiesocidae) in the Mediterranean Sea.

We report a hitherto unknown radiation within the clingfishes (Gobiesocidae), discovered in one of the best-studied marine biomes, the Mediterranean Sea. The monotypic genus Gouania is a Mediterranean endemic inhabiting the interstices of gravel beaches. Using geometric morphometric analyses, we identified two distinct morphotypes (characterized by a slender and a stout body shape, respectively) among Gouania willdenowi sampled from the three major Mediterranean basins (Eastern, Western and Adriatic). Slender and stout G. willdenowi occurred sympatrically in the Adriatic and the Eastern Mediterranean basins. Only the stout morphotype was found in the Western Mediterranean. Morphotypes were further distinguished by relative eye size and number of vertebrae. Based on mitochondrial and nuclear gene sequences, five highly divergent clades (COI-based K2P distances of 8 to 15%) were identified. The clades represented, respectively, stout Adriatic, slender Adriatic, stout Eastern, slender Eastern and stout Western Mediterranean Gouania. This suggests that the genus Gouania comprises at least five different species. Phylogenetic relationships among clades furthermore imply repeated evolution of convergent morphotypes. The onset of the Gouania radiation was dated to 3.23 (95% HPD 2.08-5.90) mya, and may therefore have overlapped with or followed the Messinian salinity crisis.

Comparative Assessment of Environmental Flow Estimation Methods in a Mediterranean Mountain River.

The ecological integrity of rivers ultimately depends on flow regime. Flow degradation is especially prominent in Mediterranean systems and assessing environmental flows in modified rivers is difficult, especially in environments with poor hydrologic monitoring and data availability. In many Mediterranean countries, which are characterized by pronounced natural variability and low summer flows, water management actions usually focus on prescribing minimum acceptable flows estimated by hydrologic methods. In this study, a comparative assessment of environmental flow estimation methods is developed in a river with poorly monitored flows and limited understanding of past reference conditions. This assessment incorporates both a hydrologic and a fish habitat simulation effort that takes into consideration hydrologic seasonality in a Greek mountainous river. The results of this study indicate that especially in data scarce regions the utilization of biotic indicators through habitat models, may provide valuable information, beyond that achievable with hydrologic methods, for developing regional environmental flow criteria. Despite the widespread use of the method, challenges in transferability of fish habitat simulation provide undefined levels of uncertainty and may require the concurrent use of different assessment tools and site-specific study.

A multilocus assessment of nuclear and mitochondrial sequence data elucidates phylogenetic relationships among European spirlins (Alburnoides, Cyprinidae).

The phylogenetic relationships and taxonomy of the spirlins in the genus Alburnoides are examined by comparative sequencing analysis of mitochondrial and nuclear markers. Molecular analyses revealed 17 Eurasian lineages divided into two main clades, termed the Ponto-Caspian and European in accordance with the lineage distribution. The indel diagnostics of ß-actin and S7 markers and translation of cyt b to the amino acid chain were evaluated as a reliable identifying tool for most of the recognised lineages. Lineage richness is closely connected with the existence of known glacial refugia in most cases. The underestimation of species richness in the genus Alburnoides is confirmed: the genetic analyses support the validity of 11 morphologically accepted species; apart from them, four phylogenetic lineages requiring descriptions as separate species were revealed. The distribution area of the nominotypical species A. bipunctatus s. stricto is newly defined. Two diverging phylogenetic lineages, A. ohridanus, and A. prespensis complex, were observed in the Southeast Adriatic Freshwater Ecoregion, confirmed as a hotspot of endemic biodiversity. A. ohridanus demonstrates high divergence from the A. prespensis complex, represented by three similar mitochondrial lineages with the same nuclear haplotypes and sympatric occurrence. The range restricted endemism was confirmed for at least seven species. The Albanian river systems, as well as the wider Ponto-Caspian basin exhibit complications among definite species delineations and gaps in understanding of microevolutionary processes; these areas require further investigations.

Phylogeny, phylogeography, and evolution in the Mediterranean region: News from a freshwater mussel (Potomida, Unionida).

The Potomida genus (Bivalvia, Unionida) has a Circum-Mediterranean distribution and like other freshwater mussel species, its populations have suffered dramatic declines. Although this genus is currently considered as monotypic, it has a long history of taxonomic revisions and presently many aspects of its systematics and evolutionary history are unclear. We sampled a total of 323 individuals from 39 different sites across the Potomida genus distribution, and sequenced two mitochondrial (16S rDNA and Cytochrome c Oxidase Subunit I) and one nuclear (28S rDNA) genes to clarify its phylogeny and phylogeographic history. Our results show that the genus includes two well-supported clades, one comprising solely the western Mediterranean species Potomida littoralis, and the other including two eastern Mediterranean species, the Greek endemic P. acarnanica and the Anatolian and Middle Eastern P. semirugata. We suggest that Potomida started radiating during the upper Miocene, and that both vicariance and dispersal events shaped the diversification and distribution of the genus along the Mediterranean region. P. littoralis is further divided in two mitochondrial lineages, one restricted to Europe and the other occurring mostly in North Africa. Moreover, some European basins present both lineages in sympatry. The conservation status of the three recognized species should be reevaluated, particularly P. acarnanica, since it is restricted to two Greek river basins presenting a high risk of extinction. Overall, our results clarify some important gaps in knowledge concerning the phylogeny, phylogeography and evolution of the Potomida genus in the Mediterranean region with important taxonomical, ecological and conservational implications.

The Gyrodactylus (Monogenea, Gyrodactylidae) parasite fauna of freshwater sand gobies (Teleostei, Gobioidei) in their centre of endemism, with description of seven new species.

While Gobioidei comprises showcases of (adaptive) radiation, the scientific interest they yielded did not ensure full understanding of goby biodiversity. Even in a well-studied region like Europe, wide knowledge gaps remain. Sand gobies represent one of the few clades whose monogenean parasites have been thoroughly studied. However, in the Balkans, part of the sand gobies' centre of endemism, these parasites were unstudied. We focus on Greek and Croatian freshwater gobies. From five sand goby species, the first parasites are reported, describing seven new Gyrodactylus species. Economidichthys pygmaeus harbours Gyrodactylus benedeni sp. n. and Gyrodactylus dorlodoti sp. n. Its congener E. trichonis hosts G. meelkopae sp. n. Knipowitschia milleri was found to host G. charon sp. n., K. thessala is infected by G. bios sp. n., and K. croatica by G. douglasadamsi sp. n. and G. hellemansi sp. n. Gyrodactylus bubyri was found on its type host K. caucasica. A diverse parasite fauna is expected for a region known for its biodiversity and endemism. The contribution of parasites to species richness in such hotspots is overlooked. The observed species richness per host is rather low compared to the better-studied eastern Atlantic sand gobies. Host vicariance is considered to mediate parasite specificity in this fauna. Some new flatworm species display unique morphological features, such as the remarkable size of the marginal hook sickle proper compared to its foot in the Economidichthys parasites, or a characteristically kinked marginal hook sickle in G. douglasadamsi sp. n. These features reflect their hosts' endemism in the Balkans.

The complete mitochondrial genome of Potomida acarnanica (Kobelt, 1879).

Freshwater mussels (Bivalvia, Unionida) play essential roles in the well-functioning of ecosystems, even providing essential services to humans. However, these bivalves face numerous threats (e.g. habitat loss and fragmentation, pollution, introduction of invasive species, and climate change) which have already led to the extinction of many populations. This underscores the need to fully characterize the biology of these species, particularly those, such as Potomida acarnanica, that are still poorly studied. This study presents the first mitogenome of P. acarnanica (Kobelt, 1879), an endemic species of Greece with a distribution limited to only two river basins. The mitochondrial genome of a P. acarnanica specimen, collected at Pamisos River (Peloponnese, Greece), was sequenced by Illumina high-throughput sequencing. This mitogenome (16,101 bp) is characterized by 13 protein-coding genes, 22 transfer RNA and 2 ribosomal RNA genes. The size of this mitogenome is within the range of another Potomida mitogenome already published for the species Potomida littoralis. In the phylogenetic inference, P. acarnanica was recovered as monophyletic with P. littoralis mitogenome in the Lamprotulini tribe, as expected. This genomic resource will assist in genetically characterizing the species, potentially benefiting future evolutionary studies and conservation efforts.

Integrating environmental DNA monitoring to inform eel (Anguilla anguilla) status in freshwaters at their easternmost range-A case study in Cyprus.

Despite significant population declines and targeted European Union regulations aimed at Anguilla anguilla conservation, little attention has been given to their status at their easternmost range. This study applies wide-scale integrated monitoring to uncover the present-day eel distribution in Cyprus' inland freshwaters. These are subject to increasing pressures from water supply requirements and dam construction, as seen throughout the Mediterranean. We applied environmental DNA metabarcoding of water samples to determine A. anguilla distribution in key freshwater catchments. In addition, we present this alongside 10 years of electrofishing/netting data. Refuge traps were also deployed to establish the timing of glass eel recruitment. These outputs are used together, alongside knowledge of the overall fish community and barriers to connectivity, to provide eel conservation and policy insights. This study confirm the presence of A. anguilla in Cyprus' inland freshwaters, with recruitment occurring in March. Eel distribution is restricted to lower elevation areas, and is negatively associated with distance from coast and barriers to connectivity. Many barriers to connectivity are identified, though eels were detected in two reservoirs upstream of dams. The overall fish community varies between freshwater habitat types. Eels are much more widespread in Cyprus than previously thought, yet mostly restricted to lowland intermittent systems. These findings make a case to reconsider the requirement for eel management plans. Environmental DNA-based data collected in 2020 indicate that "present-day" eel distribution is representative of 10-year survey trends. Suggesting that inland freshwaters may act as an unrealized refuge at A. anguilla's easternmost range. Conservation efforts in Mediterranean freshwaters should focus on improving connectivity, therefore enabling eels to access inland perennial refugia. Thus, mitigating the impact of climate change and the growing number of fragmented artificially intermittent river systems.

Ecoregions in the southern balkans: should their boundaries be revised?

Ecoregion delineations have gained increased attention in Europe, especially following the Water Framework Directive 2000/60/EC (WFD), which provides the European Union's first policy-relevant ecoregion map. However, the WFD's ecoregions were created through a minor adaptation of Illies' (Limnofauna Europaea. Gustav Fisher Verlag, Stuttgart, 1967/1978) freshwater zoogeographic regions, and the map's specific boundaries have not been widely evaluated with respect to the WFD's uses or their biogeographic accuracy. We examined the WFD ecoregion boundaries in Greece and its neighboring Balkan states by comparing them with the most prominent freshwater biogeographic boundaries as shown by riverine freshwater fish assemblages. Classification and ordination analyses of 23 river basin fish assemblages helped delineate natural faunal break boundaries in freshwater species assemblage distributions depicting major biogeographic barriers to aquatic biota dispersal. However, these biogeographic boundaries differ from those delineated in the WFD map, suggesting boundary errors and inconsistencies in the delineation method of the WFD ecoregions. We reviewed specific boundary disagreements and produced a map showing the region's most prominent freshwater biogeographic boundaries by charting them on watershed borders among the four biotically dissimilar river basin groups in the southern Balkans. This regional evaluation reveals both a need to reconcile disparate approaches to ecoregion mapping and to promote the development of a new policy-relevant inland waters ecoregion framework that would support broad-scale water management and aquatic conservation.

Assessing environmental degradation of montane riparian zones in Greece.

A survey of 218 woody vegetation plots at 109 streamside sites was undertaken to describe anthropogenic degradation of riparian zone woodlands in the mountains of mainland Greece. Two assessment indices and specific anthropogenic pressure variables were employed and they showed good correlations at both the site and river segment scales. It was demonstrated through the Spanish Riparian Forest Quality Index (Qualitat de Bosc de Riber: QBR) that most riparian zones were in moderate or good condition, although extremely few sites were of high ecological status. Most riparian sites were affected by several anthropogenic pressures, some pressures produce notable degradation beyond the site-scale (i.e. at the river segment scale). This work provides a summary of the first wide-ranging vegetation-based assessment of montane riparian zones in Greece; the results support the use of site-based rapid assessment protocols along-side aquatic ecological status surveys.

Tracking non-indigenous fishes in lotic ecosystems: Invasive patterns at different spatial scales in Greece.

Mediterranean lotic waters such as rivers, streams and springs are poorly monitored for non-indigenous fish species (NIFS). Since these systems are stressed by multiple anthropogenic pressures, it is important to build robust procedures to track NIFS distribution and spread. This study applies a multi-faceted assessment of NIFS in the lotic ecosystems of Greece at different spatial scales by providing: a) a historical review of temporal patterns and arrival pathways of fish introductions in river basins of Greece (140 basins) across 100years; b) an analysis of occurrence and abundance data of NIFS assemblages at the lotic site scale (644 electrofished sites); c) the mapping of NIFS distributional patterns at river basin (75 basins) and regional scales (7 freshwater ecoregions); and, d) a vector analysis of fish translocations using an ecoregional framework. In total, 55 NIFS were recorded (25 alien and 30 translocated); however, there is a low incidence of NIFS in lotic waters at the site scale (30 NIFS recorded in the field samples; 10 alien and 20 translocated). NIFS introductions in Greece appear to be influenced by specific socio-historical periods, indicating a gradual increase since late 1970s. Despite this increase, our study provides evidence that only four alien species are currently widespread and common in the rivers and streams of Greece: Gambusia holbrooki, Carassius gibelio, Pseudorasbora parva, and Lepomis gibbosus (in order of recorded abundance). NIFS tend to be absent or distributed in very low numbers in upland streams and in smaller river basins. However, the issue of translocated fish species is shown to be a sorely neglected problem that is difficult to track. This review tests a readily transferable screening procedure, contributes to the application of the European Union Regulation on Invasive Alien Species; it suggests gaps and uncertainties, and proposes conservation and management actions.

Integrating MAES implementation into protected area management under climate change: A fine-scale application in Greece.

Increasing anthropogenic pressures often jeopardize ecosystem integrity and policy-relevant conservation management in protected areas. To harmonize nature conservation with human well-being, EU Biodiversity Strategy to 2020 suggests Mapping and Assessment of Ecosystems and their Services (MAES) as the key concept for environmental planning and management in EU Member States. Applying this procedure is challenging due to its data-demanding and multidisciplinary nature, resulting in the ecoystem services (ES) approach being scarcely used in protected areas management. Increased data availability under EU biodiversity-related inventories and monitoring projects, as well as theoretical and empirical research advances developed during the last decade, should be put into practice to guide Member States towards local management frameworks and scenario building under the ongoing changes in the EU socio-economic environment. This study aims at filling this gap by embodying into the MAES operational framework a scenario-based approach and demonstrates this in a challenging case study of a Natura 2000 site, Lake Stymfalia, in Greece. The present management strategy, an ecological-friendly management practice, a water-efficient management practice and a non-environmentally friendly option (e.g. ecosystem destruction) are examined for current and future water demand under current and future climatic scenarios. The proposed methodological framework for ES operationalization is based on the available data (derived by EU Directives and/or modelling), expert judgment and stakeholder involvement. Therefore, this work applies and tests the importance of the MAES approach as a management and coordination platform.

Cultural landscapes and attributes of "culturalness" in protected areas: An exploratory assessment in Greece.

Cultural landscapes are poorly inventoried and evaluated in protected natural areas. This study presents a novel procedure to assess cultural landscape features and their cultural values in the major protected areas of Greece. After identifying a set of culturally modified land cover types and habitat types the GIS-based survey of the entire Natura 2000 protected area network in Greece (419 sites) shows that roughly 67% of protected area land cover consists of cultural landscape features. This was corroborated by the distribution of culturally modified habitat types which take up approximately 50% of the areal cover in a subset of the nation's Natura 2000 network (241 Special Areas for Conservation). Moreover, a set of 12 cultural attributes involving cultural heritage values, traditional land uses and aesthetic quality indicators were scored to assess these "cultural values" in each site. Gradient maps were produced to express an initial nation-wide site ranking profile. Heatmaps help link instead of solely rank culturally valuable sites that are in proximity to each other, showcasing site clusters of outstanding value. These analyses help define the level of "culturalness" of each site based on human-modified landscape and habitat types and provide a baseline review of cultural values in protected natural areas. This screening-level survey identifies the protected areas that may require special attention for managing cultural elements-of-diversity. Difficulties with data availability and uncertainties are reviewed. This procedure supports a paradigm shift that promotes a more holistic evaluation and management of biodiversity-centered protected areas, where until recently cultural landscapes were rarely appreciated.

Non-perennial Mediterranean rivers in Europe: Status, pressures, and challenges for research and management.

Non-perennial rivers and streams (NPRS) cover >50% of the global river network. They are particularly predominant in Mediterranean Europe as a result of dry climate conditions, climate change and land use development. Historically, both scientists and policy makers underestimated the importance of NRPS for nature and humans alike, mainly because they have been considered as systems of low ecological and economic value. During the past decades, diminishing water resources have increased the spatial and temporal extent of artificial NPRS as well as their exposure to multiple stressors, which threatening their ecological integrity, biodiversity and ecosystem services. In this paper, we provide a comprehensive overview of the structural and functional characteristics of NPRS in the European Mediterranean, and discuss gaps and problems in their management, concerning their typology, ecological assessment, legislative and policy protection, and incorporation in River Basin Management Plans. Because NPRS comprise highly unstable ecosystems, with strong and often unpredictable temporal and spatial variability - at least as far as it is possible to assess - we outline the future research needs required to better understand, manage and conserve them as highly valuable and sensitive ecosystems. Efficient collaborative activities among multidisciplinary research groups aiming to create innovative knowledge, water managers and policy makers are urgently needed in order to establish an appropriate methodological and legislative background. The incorporation of NPRS in EU-Med River Basin Management Plans in combination with the application of ecological flows is a first step towards enhancing NPRS management and conservation in order to effectively safeguard these highly valuable albeit threatened ecosystems.

Conservation status of freshwater mussels in Europe: state of the art and future challenges.

Freshwater mussels of the Order Unionida provide important ecosystem functions and services, yet many of their populations are in decline. We comprehensively review the status of the 16 currently recognized species in Europe, collating for the first time their life-history traits, distribution, conservation status, habitat preferences, and main threats in order to suggest future management actions. In northern, central, and eastern Europe, a relatively homogeneous species composition is found in most basins. In southern Europe, despite the lower species richness, spatially restricted species make these basins a high conservation priority. Information on freshwater mussels in Europe is unevenly distributed with considerable differences in data quality and quantity among countries and species. To make conservation more effective in the future, we suggest greater international cooperation using standardized protocols and methods to monitor and manage European freshwater mussel diversity. Such an approach will not only help conserve this vulnerable group but also, through the protection of these important organisms, will offer wider benefits to freshwater ecosystems.

Potential impacts of climate change on flow regime and fish habitat in mountain rivers of the south-western Balkans.

The climate change in the Mediterranean area is expected to have significant impacts on the aquatic ecosystems and particular in the mountain rivers and streams that often host important species such as the Salmo farioides, Karaman 1938. These impacts will most possibly affect the habitat availability for various aquatic species resulting to an essential alteration of the water requirements, either for dams or other water abstractions, in order to maintain the essential levels of ecological flow for the rivers. The main scope of this study was to assess potential climate change impacts on the hydrological patterns and typical biota for a south-western Balkan mountain river, the Acheloos. The altered flow regimes under different emission scenarios of the Intergovernmental Panel on Climate Change (IPCC) were estimated using a hydrological model and based on regional climate simulations over the study area. The Indicators of Hydrologic Alteration (IHA) methodology was then used to assess the potential streamflow alterations in the studied river due to predicted climate change conditions. A fish habitat simulation method integrating univariate habitat suitability curves and hydraulic modeling techniques were used to assess the impacts on the relationships between the aquatic biota and hydrological status utilizing a sentinel species, the West Balkan trout. The most prominent effects of the climate change scenarios depict severe flow reductions that are likely to occur especially during the summer flows, changing the duration and depressing the magnitude of the natural low flow conditions. Weighted Usable Area-flow curves indicated the limitation of suitable habitat for the native trout. Finally, this preliminary application highlighted the potential of science-based hydrological and habitat simulation approaches that are relevant to both biological quality elements (fish) and current EU Water policy to serve as efficient tools for the estimation of possible climate change impacts on the south-western Balkan river ecosystems.