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Sturgeon populations are declining worldwide and are the target of extensive conservation efforts. Addressed in several pieces of legislation, sturgeons have received considerable attention as flagship or umbrella species. Despite the need for a better understanding of the distribution and population status, the use of traditional sampling methods failed in the past, thereby hampering reliable assessments, a prerequisite for conservation. Here, we describe the development and application of an environmental DNA (eDNA) metabarcoding approach for detecting rare sturgeons in large rivers. Exemplarily, we developed a reference database for five native Danube sturgeons (Acipenser stellatus, Acipenser gueldenstaedtii, Acipenser ruthenus, Acipenser nudiventris, and Huso huso) and two non-native species (Acipenser baerii and Acipenser transmontanus), assessed these ex situ, and used eDNA as a detection tool along the entire length of the Danube (Europe, ~ 2850 km) and major tributaries. In ex situ analyses, all assays yielded positive amplifications for the assessed sturgeon species. In the Danube, the presence of A. ruthenus was confirmed at 14 of 29 sites (48.3%), and in 2 of 18 tributary sites (11.1%), providing the first comprehensive large-scale biogeographical snapshot of this species. Relative number of reads assigned to A. ruthenus varied between 0 and 2.5%, with sites registering positive detections being clustered in 3 sections of the Danube. Our findings enabled us to confirm the advantages of eDNA monitoring over traditional sampling methods for comprehensive whole-river snapshot studies of sturgeons conducted on a large geographical scale, and therefore we consider it to be a promising approach for application in conservation measures, fisheries management, scientific studies, and adaptive management plans for sturgeons on a global scale.

Vimba bream (Vimba vimba) individuals were collected from the Danube River near Belgrade, from May 2016 to May 2017. Samples of muscle and liver tissue were analyzed for concentrations of 25 elements (macro-, micro-, and toxic elements) using inductively coupled plasma optical emission spectrometry (ICP-OES). For both tissues, the highest concentrations of most elements were observed in late spring/early summer (months of May and June). Majority of statistically significant correlations between elements were positive in both tissues. Contributions of Ca, K, and Mg to human diet were in a similar range. The lowest contribution of all macroelements was observed for Na, while the contribution of Se was well above the recommended dietary allowance for this element. Levels of detected elements were well below MAC values prescribed by national and international regulations. The highest value of Hg liver/muscle index (>1) was observed in August, which could indicate a higher contamination of the sampling location in that month. Se:Hg molar ratio never drops below 1, which could indicate that Hg toxicity is prevented. THQ values showed little variation thought the year and were well below the threshold of 1, with the exception of THQ for Al in May (0.92). This element also strongly affects the hazard index.

eDNA metabarcoding is an effective method for studying fish communities but allows only an estimation of relative species abundance (density / biomass). Here, we combine metabarcoding with an estimation of the total abundance of eDNA amplified by our universal marker (teleo) using a qPCR approach to infer the absolute abundance of fish species. We carried out a 2850 km eDNA survey within the Danube catchment using a spatial integrative sampling protocol coupled with traditional electrofishing for fish biomass and density estimation. Total fish eDNA concentrations and total fish abundance were highly correlated. The correlation between eDNA concentrations per taxon and absolute specific abundance was of comparable strength when all sites were pooled and remained significant when the sites were considered separately. Furthermore, a non-linear mixed model showed that species richness was underestimated when the amount of teleo-DNA extracted from a sample was below a threshold of 0.65.106 copies of eDNA. This result, combined with the decrease in teleo-DNA concentration by several orders of magnitude with river size, highlights the need to increase sampling effort in large rivers. Our results show a comprehensive description of longitudinal changes in fish communities and underline our combined metabarcoding/qPCR approach for biomonitoring and bioassessment surveys when a rough estimate of absolute species abundance is sufficient.

This study was conducted on the Danube River locality Višnjica, exposed to the discharge of the largest wastewater collector in Serbia’s capital, Belgrade. Concentrations of metals and metalloids (Al, Fe, Cr, As, Sr, Mn, Cd, Zn, Mo, Cu, Li, Ni, B, Co, Pb, and Ba) and histopathological alterations were investigated in different tissues of common bream during one representative month of each season in 2014. This is the first study in which these two biomarkers were assessed parallelly in common bream. The highest concentrations of examined elements were noticed in gills and the lowest in muscle. Statistically significant differences in element concentrations between different seasons were noticed only in gills for Al, Cu, and Fe. Concentrations of As and Pb in fish muscle were below the maximum acceptable concentrations (MAC). The histopathological index (HI) of the liver showed higher values in comparison to the HI of the gills. Histopathological index of the gills had a significantly higher score in November in comparison to August. The liver HI had the highest score in April, and the lowest in August, while the total histopathological index had the highest score in November, and the lowest in August, both without significant differences between the months. This study endorses gills and liver as reliable organs for studying accumulation and histopathology as biomarkers of environmental changes. A faster reaction of the gills was confirmed since seasonal variations of both biomarkers were observed in this organ. Common bream proved as a good indicator of the state of organisms in polluted environments.

In complement to the JDS4 traditional fish survey (TFS, mainly by electrofishing), a fish eDNA metabarcoding-based survey has been implemented within the framework of the monitoring program organized by DNAqua-Net and in collaboration with the INTERREG "MEASURES" program. Water samples were collected at 29 sites from the source to the mouth of the Danube River, and at 18 tributaries. Two water samples (mean volume 29 L per sample) were collected at each site and the water filtered in situ. Twelve PCR replicates were performed per sample using teleo primers. In total, 80 taxa were detected, of which 19 corresponded mainly to farmed fish or food fish due to eDNA release in waste waters, most often downstream of large cities. Of the remaining 61 taxa, 50 taxa were identified at the species level, six taxa at the genus level, and five taxa at a higher taxonomic level. Concerning the Danube river itself, 69 and 57 species/markers were detected along the Danube River by TFS and eDNA surveys, respectively. 50 of these taxa were detected by both methods. Nine species were captured by TFS alone, mainly due to gaps in reference librairies and too colosely related barcodes. Eight species were only detected by eDNA. Except for the Salvelinus group, these were all benthic species, which are difficult to catch by electrofishing in large rivers (Acipenser ruthenus, Acipenser stellatus, Benthophilus sp., Romanogobio uranoscopus, Sabanejewia balcanica, Umbra krameri). The species richness tended to increase from upstream to downstream and the relative number of DNA sequences per taxa showed a clear succession of species along the river. Barbatula barbatula, Cottus gobio, Hucho hucho, Lampletra planeri, Phoxinus phoxinus and Thymallus thymallus were restricted to the Upper Danube whereas A. ruthenus, Neogobius fluviatilis, S. balcanica and Scardinius erythrophtalmus were detected from Vienna to the Danube river mouth. Abramis brama, Alburnus alburnus, Cyprinus carpio, Silurus glanis, Sander spp, Zingel streber were detected all along the river course; Alosa spp. and Syngnathus abaster downstream from the Iron Gate; A. stellatus and U. krameri only on the most downstream site (Danube delta). The calculation of a fish index, based on the common metrics used to intercalibrate national fish assessment methods on a European scale, classified most of the sites as being in moderate ecological status. Comparison of the indicative ecological status calculated using the same assessment method based on TSF and eDNA data at the common Danube sites showed a similar classification for six of the 13 sites and a difference of one class for the remaining seven sites.

This chapter examines the relationship between knowledge and Environmental Citizenship. Knowledge has been attributed to have an essential role in influencing pro-environmental behaviour and as such in the development of Environmental Citizenship. We argue that the ‘right’ kind of environmental knowledge is a necessary precondition for pro-environmental behaviour and thus for Environmental Citizenship. In this chapter, we focus on knowledge, but we also emphasise that efforts on fostering knowledge alone in education for Environmental Citizenship, without links to real life, competencies and values, are insufficient for the sake of a sustainable world. We explore which knowledge is needed in order to cultivate the coherent and adequate skills, values, attitudes and competences that we think an Environmental Citizen should have. We identify three different types of knowledge: (i) environmental systems knowledge, (ii) action-related knowledge and (iii) effectiveness knowledge. Finally, we suggest a process of co-production of new knowledge between experts and key citizens as central to the idea of a participatory approach towards developing Environmental Citizenship.

Fish erythrocytes released from erythropoietic sites are immature. During circulation, they increase their own surface, while the cell body and nuclei begin to become more elliptical and less spherical during maturation. The relative abundance of different developmental stages represents an erythron profile which could be a more sensitive indicator of contamination than classic hematological indices. In this study, we analyzed blood smears of the common nase (Chondrostoma nasus) and vimba bream (Vimba vimba) with the main goal to identify the determinants of the developmental stage of erythrocytes. Based on parameters developmental stages, erythrocytes are categorized into immature, intermediate or mature. In this investigation we have used four classification methods: the Two Step Cluster analysis, the K-Means Cluster analysis, and Neural Networks–Multilayer Perceptron and an Ordinal Regression model. Our findings clearly justify that Multilayers Perceptron and OLR models are appropriate to classify the developmental stage of fish erythrocytes in both species.

The Danube River basin represents a hotspot for the European freshwater diversity. The Danube in Serbia is characterized by a significant level of fish diversity, as well as by economically important fish resources. However, the Danube fish fauna is facing a number of adverse anthropogenic impacts, such as unsustainable fishery, habitat loss and fragmentation, water pollution and non-native species invasions. In this chapter, the major adverse impacts on the Danube fish communities in Serbia are presented, and the critical management measures are discussed, such as those related to supportive stocking and other conservation efforts. In order to ensure adequate protection and sustainable management of fish resources in the Danube in Serbia, it will be critical to introduce effective monitoring system, to establish and enforce adequate management measures, and to intensify research efforts on critical issues related to fish management and conservation. Bănăduc D, Curtean-Bănăduc A, Pedrotti F, Cianfaglione K, Akeroyd JR, editors. Human Impact on Danube Watershed Biodiversity in the XXI Century. Springer, Cham; 2020. p. 257–79. (Geobotany Studies).

This study was conducted on individuals of European perch from six reservoirs in Serbia with different properties, used for different purposes, and situated at different altitudes. Samples of muscle, gills, and liver, as well as water samples, were used for analysis of elemental accumulation. Histopathological (HP) analysis of gills and liver, and Fulton’s condition factor (CF) were used as biomarkers. Analysis of Ag, Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Li, Mg, Mn, Mo, Na, Ni, P, Pb, S, Se, Si, Sr, and Zn was conducted by inductively-coupled plasma optical emission spectrometry (ICP-OES), and accordingly, metal pollution index and bioaccumulation factor (BAF) were calculated. Elemental accumulation varied considerably both among tissues and reservoirs. Reservoirs located on large low-altitude rivers were the most heavily polluted with metals (Međuvrsje, on the Zapadna Morava River, and Lake Sava, on the Sava River). Of three tissues analyzed, the gills were most affected by pollution. The levels of Hg in muscle decreased with an increase in reservoir altitude. Specimens sampled from the highest altitude reservoir were the least affected by HP alterations; however, HP differences between the reservoirs were not statistically significant, and the severity of HP changes was generally low. No significant correlations between the elemental accumulation or HP indices with CF were observed.

Concentrations of As, Cd, Cr, Cu, Hg, Pb, and Zn in sediment samples and muscle tissue of the European perch were analyzed using inductively-coupled plasma optical emission spectrometry (ICP-OES), with the aim to assess the potential ecological (RI) and human health risk, and the degree of contamination (Cd) of three types of reservoirs in Serbia, based on their purpose: electricity generation (Vlasina, Perucac, Zaovine, and Međuvrsje), drinking water supply (Garasi), and recreation (Lake Sava). The concentrations of the studied elements were higher in sediments than in fish. However, the levels of Cd in fish caught in Vlasina, Zaovine, and Međuvrsje, Hg in Perucac and Garasi, and Pb in Lake Sava exceeded the maximum allowed concentrations. The pollution load index (PLI) indicated that sediments in all six reservoirs were contaminated, but Cd was low; a moderate contamination with Cd was observed in Vlasina, Perucac, and Zaovine, Cr in Zaovine and Međuvrsje, and Cu in Međuvrsje. A low RI was recorded for all studied reservoirs. Cd was found to be the primary contamination and ecological risk factor. Total target hazard quotient (THQ) and target carcinogenic risk factor (TR) were higher for fishers operating in these reservoirs than for the general population. Higher values of PLI, Cd, RI, and TR were observed in electricity generation reservoirs. Results indicated that this type of reservoirs suffer from higher anthropogenic pressure and/or have a worse pollution management policy compared with other types of reservoirs included in this study, especially the drinking water supply reservoir.

The life‐history traits and population dynamics of the black bullhead Ameiurus melas (Rafinesque) were studied monthly from August 2009 to December 2012 in Lake Sava in Belgrade (Serbia). This period included a mass mortality event in May 2011 and subsequent population recovery during 2012. Available evidence suggests the mass mortality was caused by European catfish virus that only affected the black bullhead. Changes in key life‐history traits and the recovery potential (increase in catch‐per‐unit‐effort, doubling of young‐of‐the‐year to adult ratio, earlier maturation and increased fecundity) following the mass mortality event suggest population resilience typical of a recurring boom‐bust pattern and demonstrate the species’ capacity to recover rapidly and re‐establish following disturbance. Repeated systematic mass removals of both juveniles and adults (using fyke nets) will be required if future fishery management measures aimed at limiting the population size are to be effective.

Due to the increasing industrialization, metals are discharged into all spheres of the environment, in particular, in river courses, which leads to the need for constant monitoring. Metals do not degrade into harmless end products; they are very persistent and have high potential for accumulation in biota. Metals in the fish body are accumulated in different amounts in the tissue specific matter. In relation to the biotic conditions and fish metabolism, the load of food, or the stage of the reproductive cycle, the seasonal variation of metal levels can be expected. Because of that, the objective of our present study was to analyze 15 metals and metalloids in liver, gills, muscle, and gonads of European chub (Squalius cephalus) throughout the 4 seasons, autumn, winter, spring, and summer. The specimens were collected from two rivers, Pestan and Beljanica at the Kolubara basin, and their concentrations were determined with inductively coupled plasma optical emission spectrometry (ICP-OES). Specimens from both rivers have shown similarities in metal accumulation like the highest accumulation of majority of elements in gills, lowest accumulation of majority of elements in muscle (except for Hg), and higher accumulation of some elements in summer (Cu, Fe, Zn). In addition, Cu and Fe showed affinity for liver, while Ba, Cr, Sr, and Zn were specific for gills. Also, Al, B, Fe, Ni, and Pb did not show significant differences in concentrations among different seasons in all investigated tissues.

Accumulation of 17 elements in muscle and liver of common nase and vimba bream, caught between February and May 2016 in the Danube River (1173 river kilometer), were assessed by ICP-OES. The principal component analysis grouped muscle and liver samples based on element concentrations (muscle grouped by higher Ba and Sr values, and liver grouped by higher Al, B, Cd, Cr, Cu, Fe, Mn, and Zn values), but no grouping between the two species was observed. Concentrations of Ba, Cu, Fe, and Zn were significantly higher in muscle, and concentrations of Ba, Cd, Cu, and Mn in liver of common nase, while vimba bream had significantly higher concentrations of Cr and Fe in liver. Common nase has a higher affinity for bioaccumulation of Cu, Fe, and Zn in muscle, while vimba bream has a higher affinity for Al, Cd, and Cr in muscle.

Metals and metalloids are natural components of the biosphere, which are not produced per se by human beings, but whose form and distribution can be affected by human activities. Like all substances, they are a contaminant if present in excess compared to background levels and/or in a form that would not normally occur in the environment. Samples of liver, gills, gonads and muscle from European chub, Squalius cephalus, were analyzed for Al, As, B, Ba, Cr, Cu, Fe, Hg, Mn, Mo, Sr and Zn using inductively coupled plasma optical emission spectrometry (ICP-OES) to highlight the importance of tissue selection in monitoring research. The comet assay or single cell gel electrophoresis (SCGE) was selected as an in vivo genotoxicity assay, a rapid and sensitive method for measuring genotoxic effects in blood, liver and gills of the European chub. Microscopic images of comets were scored using Comet IV Computer Software (Perceptive Instruments, UK). The objective of our study was to investigate two reservoirs, Zlatar and Garasi, and one river, Pestan by: (i) determining and comparing metal and metalloid concentrations in sediment, water and tissues of European chub: liver, gills, muscle and gonads (ii) comparing these findings with genotoxicity of water expressed through DNA damage of fish tissues. A clear link between the level of metals in water, sediment and tissues and between metal and genotoxicity levels at examined sites was not found. This suggests that other xenobiotics (possibly the organic compounds), contribute to DNA damage.

The concentrations of As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn in the muscle, gills, liver and intestine of the wels catfish (Silurus glanis) from the Danube River were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The aim of the study was to determine whether in complex muscle/skin, gill filament/gill arch, proximal/distal liver and proximal/median/distal intestine samples, particular components differ in concentrations of the analyzed elements. Results indicated that there were no differences in the accumulation of different elements between the proximal and distal liver segments and between the proximal and median intestine sections. Conversely, elemental accumulation patterns in muscle and skin differed significantly. Significant differences were also observed between the gill arch and filaments, as well as between the distal and the two upper intestine sections. Findings indicated the importance of detailed reporting of tissue sampling, i.e. whether the skin was included in the muscle sample, as well as if the gill arch and filaments were analyzed together. Due to a potential bias that can be produced by different muscle/skin or gill arch/filament ratios included in the sample, we strongly recommend that they should not be analyzed together. Results of the present study might be of interest to the scientific community and stakeholders involved in aquatic ecosystem monitoring programs. [Projekat Ministarstva nauke Republike Srbije, br. TR37009 i br. 173045]

Chub (Squalius cephalus) specimens were collected in Korenita River seven months after spillover from the waste water of antimony mine tailing pond and compared with chub living in Kruščica reservoir (intended for water supply) and Međuvršje reservoir (influenced by intense emission of industrial, urban and rural wastewater). Concentrations of 15 elements (Al, As, B, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Si, Sr, Zn) were determined in muscle, liver and gills of chub by inductively coupled plasma optical spectrometry (ICP-OES) and histopathological alterations in liver and gills were assessed. Chub specimens from Korenita River had higher concentrations of As, Ba and Pb in all three investigated tissues as well as higher total histopathological index values than chub from reservoirs. Specimens from Međuvršje reservoir were characterized by higher values for concentrations of Cu and Si in muscle tissue and higher values for regressive histopathological alterations in gills. Individuals of chub from Kruščica reservoir had the highest concentrations of Fe in liver, Hg in muscle and Sr and Zn in muscle while gills had the lowest value of total histopathological index. The results from the present study showed higher level of histopathological alterations as a result of mine tailing accident. As a result of mixed contamination on the Međuvršje site, histopathological index values of gills were in line with the index value from Korenita River. Increased values for Fe and Sr in chub tissue from Kruščica reservoir could be explained by geological structure of the site which is characterized by magmatic rock rich in Cu, Fe and Ni as well as dominant carbonate sediment complex of marine origin with increased level of Sr.

The major objective of this study was to investigate the impact of wastewaters on the level of genotoxicity and the possible reflection of genotoxicity on the genetic variability in chub (Squalius cephalus) collected from the sites under different pollution pressures along the Sava River. To emphasize the impact of wastewaters, this study was carried out in a season with low water discharge. The level of genotoxic potential at the sites was studied by alkaline and Fpg- modified comet assay in fish blood, while RAPD technique was employed on the fish muscle tissue. The variations in the genotoxic potential were observed along the Sava River based on the level of DNA damage in blood of the specimens collected at the selected sites. Clustering of the impacted sites was also observed based on RAPD fingerprints. Comparison of the data obtained in RAPD and the data from the comet assay indicated the possible reflection of genotoxicity on the genetic variability in chub collected from the sites with different pollution pressure. This is a preliminary study which reports the differentiation of the population of chub along the Sava River based on RAPD analysis.

The paddlefish (Polyodon spathula) was first introduced to Europe in 1974, mainly due to its potential for rearing in natural polyculture ponds and large temperate reservoirs. The information on the history of paddlefish aquaculture efforts in Europe is scarce, as well as data on current paddlefish aquaculture status and trends. In addition, there is a lack of data on its presence and potential establishment in the wild, while its invasive potential and associated risks and impacts are largely unknown. In order to evaluate its current status in Europe, we conducted a survey among scientists, aquaculture producers and other stakeholders, and reviewed literature and data on the Internet. Based on the results obtained, we discuss the potential and the challenges in European paddlefish aquaculture development, and analyze paddlefish invasive potential and risks associated with its naturalization. Paddlefish aquaculture is well established only regionally in Europe, but offers relatively high potential for further development in pond farms. Nevertheless, future development will require careful planning, especially regarding market development and improved marketing strategies. While paddlefish likely represents a low-risk invader, improved control and reporting on trade and intentional and unintentional releases will be required. Given the lack of knowledge on potential impacts following its introduction, due caution seems highly advisable. This is the peer reviewed version of the following article: Jarić I, Bronzi P, Cvijanović G, Lenhardt M, Smederevac-Lalić M, Gessner J. Paddlefish (Polyodon spathula) in Europe: An aquaculture species and a potential invader. J Appl Ichthyol. 2018, which has been published in final form at [http://doi.org/10.1111/jai.13672]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.

Metal content was evaluated in the sediment, macrophytes and fish in the Medjuvrsje reservoir (Western Serbia). Concentrations of 16 trace elements (Ag; Al; As; B; Ba; Cd; Co; Cr; Cu; Fe; Li; Mn; Ni; Pb; Sr; Zn) were analysed in the sediment, macrophytes and fish of an aquatic ecosystem. Five macrophyte species and three fish tissues (liver, muscle, gills) from five fish species (freshwater bream, common nase, Prussian carp, chub, wels catfish) were sampled and the metal content was analysed with ICP-OES. The sediment concentrations of Cu, Cd, and Zn exceeded the Canadian sediment quality guidelines while concentrations of Cr and Ni were above the Netherlands’ target values. Bioaccumulation factors (BSAF) were calculated for analysed macrophytes and fish tissue. The BSAF had higher values for macrophytes for all investigated elements except for Cu and Zn; Cu had a higher value in the liver of the freshwater bream (0.823) and Zn had a higher value in the liver of freshwater bream (0.914) and chub (0.834) as well as in gills of Prussian carp (2.58) and chub (1.26). Potamogeton pectinatus, Ceratophylum demersum and the root of Phragmites communis showed higher accumulation of elements than Trapa natans and Potamogeton fluitans and the body of P. communis. The highest BSAF values for Ba, Mn, Sr and Ni were recorded in the gills. Cd and Cu had the highest BSAF values in the liver. Results confirmed that particular macrophyte and fish species could be a good indicator of reservoir water and sediment pollution.

Population models in fish represent one of the major scientific approaches to identify and bridge existing gaps in the understanding of the ecology and life history, as well as to support management. In general, the more complex the models get, the more they are restricted to a case-by-case basis for particular, well-studied species, because demographic data required for detailed models are unavailable for the majority of species. In the present study, we propose a simple life-table metamodel, which facilitates population and fishery assessments across entire groups of fish. The general approach is described and its application is demonstrated on two groups of fish, which reflect extremes in their life cycle: sturgeons (order Acipenseriformes) and shads (genus Alosa). The approach allows to determine fishing mortality thresholds across different life history types, and to analyze their general population elasticity. Its application is suggested for rapid assessments across large, species-rich groups of fish. The method also allows utilizing life history data from well-studied species to infer fishing mortality thresholds for other, poorly studied species within the same group. Comparisons of the model output with results from other population models indicated a good congruity.

The main objective of this study was to evaluate the concentrations and seasonal variations of trace elements in surface sediments of three major rivers in Serbia—the Danube, the Zapadna Morava (ZM), and the Južna Morava (JM)—according to sediment quality guidelines. The ZM and the JM create the Velika Morava River, one of the most important tributaries of the Danube, which has been characterized as a source of heavy metal pollution. The total concentrations of 15 elements (Al, As, B, Ba, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sr, and Zn) were determined in surface sediments (0–15 cm depth) collected during three seasons using inductively coupled plasma spectroscopy (ICP-OES). Principle component analysis (PCA) was used to identify the main variations in metal concentrations and grain size distribution. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM-EDS) was used for grain analysis. PCA and three-way MANOVA results showed significant differences in element concentrations and grain size distribution between the rivers, and significant seasonal differences for each river. The concentrations of Cu and Ni exceeded sediment quality guideline levels in the ZM and the Danube, respectively, while excess Hg was detected in all three rivers. Concentrations of Al, Ba, Cu, Fe, Sr, and Zn significantly varied between seasons in the Danube and the ZM, being the highest in the summer. In the JM, concentrations of Al, As, Fe, Mn, and Zn varied with season, with the lowest values in the summer. The ZM had the highest percentage of silt and clay, and SEM-EDS analysis of ZM sediments showed associations of Cu with carbonate hydroxides and/or iron oxides in particles

Poor regulated fishery, pollution, fragmentation and loss of habitat are most important factors influencing decline of sterlet population worldwide. In Middle and Lower Danube region, this species still have significant economic importance since wilde populations are commercially exploited, while Upper Danube populations are dependent on stocking efforts in order to maintain their presence in open waters. Aim of present study is to analyze genetic diversity of sterlet populations from the Middle and Lower Danube and Lower Tisza rivers, as a prerequisite for their effective conservation and management. Analysis of a highly variable D-loop fragment of mitochondrial DNA detected five new haplotypes, while the eight previously identified haplotypes had extended their previous range. Genetic variability could be attributed almost entirely to individuals, with observed lack of population structure. Negative values of neutrality test indicate recent expansion on some sampling locations. Adittionaly, gene flow analysis between Lower and Middle Danube region showed intensive exchange of speciemens. At the same time analysis showed some influence of Tisza dam on gene flow between samples from Tisza and Middle Danube section. Our study indicated the need for a careful planning of sterlet stocking programmes and inclusion of demographic data or catch time-series. Ministry of Education, Science and Technological Development of the Republic Serbia {[}173045]

The effects of physical factors on fish catch in the Serbian part of the Danube River were studied for period of six decades. The data on total catch for the Danube River from river kilometre 845 to river kilometre 1433 were collected from Statistical Office of the Republic of Serbia, while water level and water temperature data were collected from 16 water gauge stations along the investigated part of the Danube River for the period 1948–2009. Cross-correlation functions have been used to analyse the functional connection between Danube water level, water temperature and fish catch while ARMA model which combines cyclic (deterministic) and random (stochastic) components of the analysed sequences was used for the forecasts. The cross-correlation function showed negative correlation between water level and temperature as well as between water temperature and catch and positive correlation between water level and catch. The Danube water level and catch were coherent at the periods of 2.06, 4.13, 6.2, 10.33, 20.66 years, while the cross correlation function between these time series did not show phase lag. The results of reconstruction and forecast of water level, temperature, and catch of fish in the Danube River, obtained by summing the cyclic and stochastic components, was used for the forecast till 2029. In 2016, seven years after, the initial forecasts were made, validity of the model was checked by obtaining data for water temperature, water level and fish catch in the Danube River for the period 2010–2015. Model gave the best prediction for water temperature; average standard error was 1.6 times higher for predicted value than for model value while for fish catch and water level they were 1.96 and 4.97, respectively. Methods used in this work could be powerful tool for prediction of fish catch and serve as the basis for better fisheries management. Science of The Total Environment (2017), 609: 664-671

Biomarkers attract increasing attention in environmental studies, as a tool for detection of exposure and effects of pollution, from both natural and anthropogenic sources. This study aims to assess the impact of multiple stressors during distinctive seasons, covering also extreme hydrological events (extensive flooding in the mid May 2014), on different levels of biological organization in the liver and gills of three closely related freshwater breams. Our previous study on DNA damage in blood cells of these specimens showed increased DNA damage in June 2014, one month after the flooding event. As a continuation of that research, the present study was conducted. As a biomarker of exposure DNA damage was measured by applying the alkaline comet assay, while histopathological alterations were monitored as a biomarker of effect. Additionally, concentrations of metals and metalloids in gills, liver and muscle were assessed. Sampling of fish tissues was performed in 2014, during winter (January and February), spring (March and early June) and summer (late June, July and August). Significant seasonal difference in DNA damage was observed for both tissues. During spring and summer the level of DNA damage in gills was significantly higher when compared to the liver. Histopathological analyses showed higher frequency of alterations in gills during spring, and in liver during summer, but without a significant seasonal difference. Gills had the highest concentration of metals and metalloids during the spring and summer, and liver during winter. Muscle was the least affected tissue during all three seasons. This study highlighted the importance of the multiple biomarker approach and the use of different fish tissues in assessment of surface water pollution.

Studies of metal accumulation in fish are mainly focused on the muscle tissue, while the metal accumulation patterns in other tissues have been largely neglected. Muscle is not always a good indicator of the whole fish body contamination. Elemental accumulation in many fish tissues and organs and their potential use in monitoring programs have not received proper attention. In the present study, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, and Zn concentrations were assessed by inductively coupled plasma mass spectrometry (ICP-MS) in the following 14 tissues of the wels catfish (Silurus glanis) from the Danube River: muscle, gills, spleen, liver, kidneys, intestine, gizzard, heart, brain, gallbladder, swim bladder, vertebra, operculum, and gonads. A high level of differential elemental accumulation among the studied tissues was observed. The maximum overall metal accumulation was observed in the vertebra, followed by the kidneys and liver, with the metal pollution index (MPI) values of 0.26, 0.25, and 0.24, respectively. The minimum values were observed in the gallbladder, muscle, brain, and swim bladder, with MPI values of 0.03, 0.06, 0.07, and 0.09, respectively. Average metal concentrations in the fish muscle were below the maximum allowed concentrations for human consumption. The mean As, Cd, Pb, Cu, Fe, and Zn concentrations in the muscle were 0.028, 0.001, 0.001, 0.192, 3.966, and 3.969 mu g/g wet weight, respectively. We believe that the presented findings could be of interest for the scientific community and freshwater ecosystem managers. There is a need for further research that would assess less studied tissues in different fish species.

Invasive species have the ability to modify their life-history traits in newly colonized areas, with positive shifts in specific life history traits under favourable environmental conditions. If such positive changes in their life history result in a comparably larger population growth rate, it may give them a competitive edge over native species, support faster range expansion and contribute to their invasion success. Within the present paper we hypothesized that the demographic flexibility represents an important contribution to the invasion success of exotic species, and that demographic flexibility patterns of invasive species differ from those in unsuccessful invaders. We tested this hypothesis by the use of elasticity analysis applied on simple age-structured population models of invasive fish species in the Danube River, as well as of non-native species that failed to establish or become invasive. Findings imply that the invasive fish species could have the ability to experience a more rapid population growth under favourable environmental conditions, especially those that sustain recruitment, while at the same time being more robust to changes in survival. The highest population elasticity among the assessed alien invasive species was detected in stone moroko (Pseudorasbora parva). The described approach has the potential to be used as an additional screening tool for invasive species. When combined with other invasion risk profiling methods, it can provide additional insight into characteristics of species invasions and in invasion potential of a species.

Emerging scientific fields are commonly identified by different citation based bibliometric parameters. However, their main shortcoming is the existence of a time lag needed for a publication to receive citations. In the present study, we assessed the relationship between the age of references in scientific publications and the change in publication rate within a research field. Two indices based on the age of references are presented, the relative age of references and the ratio of references published during the preceding 2 years, and applied thereafter on four datasets from the previously published studies, which assessed eutrophication research, sturgeon research, fisheries research, and the general field of ecology. We observed a consistent pattern that the emerging research topics had a lower median age of references and a higher ratio of references published in the preceding 2 years than their respective general research fields. The main advantage of indices based on the age of references is that they are not influenced by a time lag, and as such they are able to provide insight into current scientific trends. The best potential of the presented indices is to use them combined with other approaches, as each one can reveal different aspects and properties of the assessed data, and provide validation of the obtained results. Their use should be however assessed further before they are employed as standard tools by scientists, science managers and policy makers.

Pikeperch (Sander lucioperca), European catfish (Silurus glanis), burbot (Lota lota), and common carp (Cyprinus carpio) were collected from the Danube River (Belgrade section, Serbia), and samples of liver, muscle, and gills were analyzed for Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Hg, Li, Mn, Mo, Ni, Pb, Se, Sr, and Zn using inductively coupled plasma optical emission spectrometry (ICP-OES) to highlight the importance of species and tissue selection in monitoring research, contaminant studies, and human health research. The Kruskal-Wallis test revealed significant differences between fish species in regard to metal levels in liver, muscle, and gills. The principal component analysis (PCA) indicated that the studied fish species could be grouped on the basis of the level of analyzed elements in liver and gills. The Mann-Whitney test showed two subsets (one comprising two piscivorous species, pikeperch and catfish, and the other, two polyphagous species, burbot and carp) in regard to Cr and Hg levels in liver (higher levels in piscivorous species), as well as B, Fe, and Hg in gills (B and Fe with higher levels in polyphagous and Hg in piscivorous species), and As in muscle (higher levels in polyphagous species). Carp had distinctly higher levels of Cd, Cu, and Zn in liver in comparison to other three species. None of the elements exceeded the maximum acceptable concentrations (MAC). However, since Hg levels are close to the prescribed MAC levels, the consumption of these fishes can be potentially hazardous for humans.

Anadromous fish could have intraspecific groups with different life histories; identification of these is very important for management and conservation. Geometric morphometrics represents a quick and cost-effective method to identify such intraspecific groups. In this study, geometric morphometrics of Pontic shad (Alosa immaculata) migrants from the Danube River were investigated, as was Fulton's condition factor (K), hepatosomatic index (HSI) and gonadosomatic index (GSI). Pontic shad specimens were collected during April and May of 2004 and 2006. The sample comprised four groups (65 males and 56 females in 2004; 110 males and 78 females in 2006). Geometric morphometrics revealed differences in shape that were statistically significant for both year (nested analysis of variance: F = 13.84; p < 0.001) and sex (nested analysis of variance: sex nested in year, F = 1.71; p < 0.01). The major differences between years were in head shape and position of fins. The most significant difference between females an...

Pikeperch (Sander lucioperca), European catfish (Silurus glanis), common carp (Cyprinus carpio), and gobies (Neogobius gymnotrachelus, Neogobius melanostomus) were collected from the Danube River (Belgrade section), and samples of liver, muscle, or whole-body composites (in the case of gobies) were analyzed for As, Cu, Fe, Hg, Mn, and Zn with inductively coupled plasma optical spectrometry to find out if there was a correlation between accumulation of these elements in predatory and prey species, as well as in pairs of species with overlapping diets. Concentrations of all analyzed elements were either higher (Cu, Fe, Mn, Zn) in liver than in muscle, or equal (As, Hg), except for Hg in carp, which was higher in muscle. Mercury concentration in liver and muscle of predators (catfish, pikeperch) was significantly (

Recent technological developments have increased the number of variables being monitored in lakes and reservoirs using automatic high frequency monitoring (AHFM). However, design of AHFM systems and posterior data handling and interpretation are currently being developed on a site-by-site and issue-by-issue basis with minimal standardization of protocols or knowledge sharing. As a result, many deployments become short-lived or underutilized, and many new scientific developments that are potentially useful for water management and environmental legislation remain underexplored. This Critical Review bridges scientific uses of AHFM with their applications by providing an overview of the current AHFM capabilities, together with examples of successful applications. We review the use of AHFM for maximizing the provision of ecosystem services supplied, by lakes and reservoirs (consumptive and non consumptive uses, food production, and recreation), and for reporting lake status in the EU Water Framework Directive. We also highlight critical issues to enhance the application of AHFM, and suggest the establishment of appropriate networks to facilitate knowledge sharing and technological transfer between potential users. Finally, we give advice on how modern sensor technology can successfully be applied on a larger scale to the management of lakes and reservoirs and maximize the ecosystem services they provide.

This study examines the use of freshwater bream (Abramis brama) as a sentinel organism for genotoxicity assessment of the Danube River using the comet assay. Sampling of bream was performed during February, April, August, and November in 2014 to assess seasonal variation of DNA damage level as a response to genotoxicity in annual cycle. Additionally, concentrations of fecal coliforms and enterococci were analyzed and they indicated a critical to strong level of fecal pollution on investigated locality during annual cycle. Comet assay was performed on blood, liver, and gill cells of bream. DNA damage level was expressed using tail intensity (TI %), Olive tail moment (OTM), and tail length (TL pix). According to TI and OTM, all three tissues had the highest level of DNA damage in August. The lowest level of DNA damage in liver was measured during February, in blood during November, and in gills during April. According to TL, gills had the highest level of DNA damage in February, and liver cells had the lowest level of damage during April. Multiple correspondence analysis (MCA) showed that DNA damage in blood cells is under the strong influence of variations in NO2, NO3-, NH4+ levels and also the variation in temperature and oxygen levels. DNA damage in liver cells is highly associated with the variations of Mn, Fe, Cu, Zn, and PO43- levels. DNA damage in gill cells is strongly affected by the variations of As, Cd, Pb, Cr, and COD (Mn) levels. Freshwater bream is shown to be a potentially good indicator organism in genotoxic potential field studies.

In this study, 20 heavy metals and trace elements (Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Si, Se, Sr and Zn) were analyzed in different tissues (muscle, liver, intestine, feather and bone) of the great cormorant (Phalacrocorax carbo) and in different tissues (muscle, liver, gills, kidney and gonad) of their main prey (common carp—Cyprinus carpio and Prussian carp—Carassius gibelio) during the nesting season. Cormorant and fish specimens were collected at the Ecka Fishing Farm (Serbia) and from the nearby river. Principal components analysis (PCA) showed that cormorant livers were differentiated from other four tissues by higher concentrations of Hg, Fe, Cu and Mo, feathers by higher concentrations of Al and Si, and bones by higher concentrations of Sr and Mg. Differentiation among the three age classes of cormorants was observed only with regard to elemental concentrations in their feathers: subadult cormorants were differentiated by higher Hg and Zn concentrations, while the adults were separated by higher concentrations of Fe. In comparison with their prey (common carp and Prussian carp) cormorants were differentiated by higher concentrations of Fe, Hg and Cu in the muscle. Toxic mercury concentrations above 4 μg g−1 dw were determined in the liver of juvenile cormorants, as well as in the liver and feathers of subadults. Hg concentrations were significantly correlated among all studied tissues. Cormorant had significantly higher concentrations of Fe and Hg in muscle and liver than two studied fish species.

Summary The aim of this paper was to use geometric morphometry as a helpful tool to clarify the overlap between meristic characters (such as the number of anal fin rays) of different fish identification keys employed for the two sister species, Ameiurus melas and Ameiurus nebulosus. Geometric morphometry included shape analysis of 15 landmarks on 432 specimens. The analyses showed significant differences among groups based on the sampling site, while there was no difference among groups based on the number of anal fin rays (used to distinguish species). This suggests that the use of meristic characters for the determination of these two sister species is rather questionable.

Summary This paper presents the status and trends in management of sturgeon species and the development of sturgeon aquaculture in the Lower Danube countries: Romania, Bulgaria, Serbia, Ukraine and Moldova. Sturgeon fishery moratoria and aquaculture development represent first steps in the Lower Danube countries to combat extirpation. Supportive stocking was used as a compensation for the impact of sturgeon fishery and dam construction, but these efforts unfortunately lacked adequate cooperation and coordination between and among region countries. Unsolved problems remain, such as the presence of illegal sturgeon fishery, water pollution, habitat destruction and fragmentation. Construction of fish passes and habitat restoration project developments are two key issues that have yet to be tackled in the Lower Danube region.

Data on the presence of invasive species are often scarce, especially during the initial period following their introduction, when population abundance and species detectability are both still very low. In such cases, sporadic sightings in their newly occupied territory are often the only available information about their presence and distribution. Here, we demonstrate the potential of using probabilistic models for the assessment of the distribution of newly established invasive species in rivers, based on their sighting records. We have applied Solow’s equation to the sighting record of the Amur sleeper (Perccottus glenii) in the Danube River, to assess the probable range of its population. The method applied indicated that, at the time when the sightings have been made (2003–2008), the Amur sleeper population in the Danube most probably extended between the 704.5 and 1,178.5 river km, while it was highly improbable (P < 0.05) that it was present downstream from the 606 river km or upstream from the 1,277 river km. The application of this approach has, however, to be tested further on a larger number of scenarios and case studies in order to evaluate its reliability. What this method lacks in complexity, compared to more advanced species distribution assessments, it makes up by its ability to be easily used and quickly applied for preliminary assessments of the ranges of riverine species. As a result, the proposed approach might be of interest for scientists and managers dealing with invasive species.

The accumulation of heavy metals in fish has been extensively studied and well documented. However, the research has been mainly focused on the muscle tissue, while the distribution patterns among other tissues, such as liver and gills, have been mostly neglected. Within the present study, the concentrations of Ag, Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, Sr, Zn and Li in the muscle, gills, liver and intestine of the sterlet (Acipenser ruthenus) from the Danube River have been assessed by using the inductively-coupled plasma optical spectrometry. The analysis has revealed a high degree of differential accumulation of the studied heavy metals and trace elements in the tissues of the Danube sterlet. Liver appeared to be the main heavy metal storage tissue, while the lowest levels of analyzed metals were found in muscles. Heavy metal concentrations in the muscle were at acceptable levels for human consumption, except partly for cadmium. We conclude that, in order to enable a more efficient control of contaminants in fish products, there is an apparent need to resolve the problem of lack of legal limits for many metals in European legislation.

A total of 22 fish species have been introduced into the inland waters of Serbia, either intentionally or accidentally. This paper provides a summary of data concerning time and reason of introduction, mode of expansion, degree of acclimatization, impact on native fish and estimated area of recent distribution. Four of the non-native fish species currently occupy more than 51% of Serbian territory while 5 of them occupy between 21-50% of territory. This paper reviews impacts of introduced freshwater fish in Serbia based on collected data.

Sturgeon populations in the Danube River have experienced severe decline during the last several decades, mostly due to the poorly regulated fishery, river fragmentation and water pollution. This study focuses on gaining better understanding of sturgeon life history primarily by addressing the assessment of microelement accumulation in sturgeon pectoral fin rays, especially of strontium and calcium, as a method that can reveal migration patterns of anadromous sturgeons. Analysis was performed on pectoral fin samples of three anadromous Danube sturgeon species (beluga, Russian sturgeon and stellate sturgeon) by the use of a Nuclear Microprobe technique. The most frequent pattern in analyzed samples was represented by a low Sr:Ca ratio in the innermost annuli, followed by an increased ratio in the middle annuli segment, and often with a decreased ratio in the outermost annuli. Probability density estimate has revealed three distinguished maxima of the Sr:Ca ratio, 7.08×10 �3 , 8.98×10 �3 and 9.90× 10 �3 , which might correspond, respectively, to fresh, brackish and saltwater. Although the analysis of the Sr:Ca ratio in sturgeon pectoral fin rays has revealed changes that might indicate probable migration between habitats with different water salinity, further studies are needed for improvement of this method. This study represents the first analysis of this kind that was conducted on sturgeon species from the Black Sea basin.

Pontic shad ( Alosa immaculata Bennet 1835) is an anadromous species that lives in the heavily polluted north-western part of the Black Sea and migrates into the Danube River to spawn. To assess their heavy metal contamination levels, samples of Pontic shad were collected at 863 river kilometre of the Danube River. Muscle, liver and gill samples were prepared using microwave digestion, and the analysis of Al, As, Cd, Cu, B, Ba, Fe, Mg, Sr, Zn, Li, Co, Cr, Mn, Mo, Ni and Pb was performed with inductively-coupled plasma-optic emission spectroscopy (ICP-OES). Significant differences in concentrations of analyzed elements were observed among different tissues, as well as between the genders. Al, Sr, Ba, Mg, and Li had the highest concentrations in gills, while Cd, Cu, Zn, Fe and B were highest in the liver. While the muscle had the lowest concentrations of most of the analyzed elements, it had the highest concentration of As. This study has revealed that the concentrations of Cd and As in Pontic shad muscle tissue were above the maximum acceptable concentrations for human consumption, thus precautions need to be taken in order to prevent future heavy metal contamination.

Construction of the dams in Lower Danube River (LDR- rkm 0 Sulina – rkm 1075 Baziasi) without fish passages, as well as illegal fishery and river bottom modification due to navigation had negative impact on sturgeon’s populations. Beluga Sturgeon, Stellate Sturgeon and Russian Sturgeon still migrate to spawn in LDR but there is lack of information relating to sturgeons long distance migration in this area. Tagging with Vemco V16 TP acoustic tags, equipped with temperature and depth sensors involved 36 adult Beluga and 44 Stellate Sturgeons provide new data about sturgeon migration. Their migrations are monitored by VR2W receivers, deployed on between rkm 71 and rkm 864. Analysis of data showed that 17% of Beluga and 30% of Stellate Sturgeon never recorded and 30% and 11% of Beluga and Stellate Sturgeon recorded in period more than 30 days after tagging. Only 5% of Beluga and 7% of Stellate Sturgeon have been recorded after a period of more than 1 year. During 2013-2016 4 adult Beluga and 1 adult Stellate Sturgeon were recorded in the IGII Dam area. Results of this investigation showed that sturgeons still approached the IG2 dam and construction of fish passage on this dam is needed.

Summary A sample of 198 asp (Aspius aspius) was collected over a 28.5 km long section of the Danube River, upstream and downstream from the mouth of the Sava River, in order to provide information on population structure, diet, growth, and condition of this species. Total body lengths of sampled fish were 90–405 mm, and body weights 5–567 g. The sample was predominantly sub-adult fish aged 0+ to 4+ years. Sexually immature asp prevailed over the sexually mature asp. The sex ratio (1.4 : 1) was biased toward males. Male asps in the Danube become sexually mature only in their fifth year of life, and the females even later, as no sexually mature females were caught. The diet spectrum consists exclusively of fishes; of seven determined prey species, most important are bleak Alburnus alburnus, roach Rutilus rutilus, and goldfish Carassius auratus. Highest value of the seasonal vacuity index is in November (Iv = 94.1), and the smallest in July (Iv = 35.6). Trophic niche breadth is largest in May (H = 1.40), and smallest in November (H = 0.00). The largest average number of prey items is found in age class 4+ (2.33), the smallest in age class 0+ (1.00). Von Bertalanffy growth parameters are L∞ = 526.46, K = 0.28, and t0 = 0.22. The length–weight relationship is described as log W = −5.16 + 3.03 log L. Fulton’s condition factor increases with fish length, from 0.70 to 0.83. Regarding seasonal fluctuations, this factor begins to increase in late spring, with the trend continuing until autumn, and then decreasing from autumn to spring.