Assessment of trace and macroelement accumulation in cyprinid juveniles as bioindicators of aquatic pollution: effects of diets and habitat preferences.

Juveniles of three cyprinids with various diets and habitat preferences were collected from the Szamos River (Hungary) during a period of pollution in November 2013: the herbivorous, benthic nase (Chondrostoma nasus), the benthivorous, benthic barbel (Barbus barbus), and the omnivorous, pelagic chub (Squalius cephalus). Our study aimed to assess the accumulation of these elements across species with varying diets and habitat preferences, as well as their potential role in biomonitoring efforts. The Ca, K, Mg, Na, Cd, Cr, Cu, Fe, Mn, Pb, Sr, and Zn concentration was analyzed in muscle, gills, and liver using MP-AES. The muscle and gill concentrations of Cr, Cu, Fe, and Zn increased with trophic level. At the same time, several differences were found among the trace element patterns related to habitat preferences. The trace elements, including Cd, Pb, and Zn, which exceeded threshold concentrations in the water, exhibited higher accumulations mainly in the muscle and gills of the pelagic chub. Furthermore, the elevated concentrations of trace elements in sediments (Cr, Cu, Mn) demonstrated higher accumulation in the benthic nase and barbel. Our findings show habitat preference as a key factor in juvenile bioindicator capability, advocating for the simultaneous use of pelagic and benthic juveniles to assess water and sediment pollution status.

Freshwater fish scale database.

Scales of the ray finned fishes can be used for multiple purposes. Beside others by their specific structure and elemental composition they are usable for age determination and food-web researches. Additionally, just their presence, absence, shape, location, or numbers could provide a reliable taxonomic information. The fish scales show remarkable size variation also, which characteristics provide reliable information about the environmental needs of freshwater fish. But till now this information was not interpretable and comparable in the absence of a sufficiently large and detailed database. In this study we provide a database which can facilitate the further complex comparative studies. Our dataset consists of 2954 scale photos characterising 193 freshwater fish species. All photos have a uniquely identification code showing exactly which species the scale belongs to. In addition to the photos, our database includes a table showing the exact taxonomic classification of the studied species, the average body profile-index, and the relative scale size, as well as the ecological (flow and feeding habitat preferences) and life strategic characteristics (feeding and breeding guild memberships) of each species. The sampled species have diverse origin, covering five biogeographical regions. An average of five adults from each species were selected for sampling. Three-to-four scales were collected from the anterior part of the body between the dorsal fin and the lateral line. The removed scales were cleaned and prepared to microscope slides, and after this process they were photographed or scanned. Our database can be used for both research and educational purposes. A large number of scale photos assigned of species can be an opportunity to create an automatic species identification system. Moreover, the subjectivity in morphometric measurements can be eliminated by analysing the database with artificial intelligence.

eDNA metabarcoding reveals the role of habitat specialization and spatial and environmental variability in shaping diversity patterns of fish metacommunities.

Information is scarce on how environmental and dispersal processes interact with biological features of the organisms, such as their habitat affinity, to influence patterns in biodiversity. We examined the role of habitat specialist vs. generalist species, and the spatial configuration, connectivity, and different environmental characteristics of river-floodplain habitats to get a more mechanistic understanding of alpha and beta diversity of fish metacommunities. We used environmental DNA metabarcoding to characterize species (taxa) richness and composition in two separate floodplains of the river Danube (Austria and Hungary) during two different hydrological conditions. Results showed that differences in the number of generalist and specialist species and their responses to connectivity and environmental gradients influenced patterns in alpha and beta diversity. Of the components of beta diversity, richness difference (nestedness) showed consistently higher values than replacement (turnover), mainly due to the decrease of specialist species along the connectivity gradient (i.e., from the mainstem to the most isolated oxbows). Variance in both alpha and beta diversity could be well predicted by a set of local and regional variables, despite high environmental variability, which characterizes river-floodplain ecosystems. Of these, the joint or shared variance fractions proved to be the most important, which indicates that the effects of local and regional processes cannot be unambiguously separated in these river-floodplain systems. Local scale environmental variables were more important determinants of both alpha and beta diversity in the low water period than in the high water period. These results indicate the differential role of local and regional processes in community organization during different hydrological conditions. Maintenance of both local and regional scale processes are thus important in the preservation of alpha and beta diversity of floodplain fish metacommunities, which should be considered by environmental management.

Hydrology is a major influence on amphibian abundance in a large European floodplain.

River-floodplain ecosystems play a crucial role in connecting landscape patches through hydrological connectivity, but they are among the most threatened ecosystems. Floodplains provide important habitat for amphibians by connecting aquatic and terrestrial habitats. Modifications to floodplain hydrology can impact amphibian communities, yet few studies have examined amphibian metacommunities in floodplain wetlands.In this study, we assessed patterns in amphibian breeding abundance in one of the largest floodplains of the Danube River, Hungary, relative to hydrological connectivity and multi-scale variables at 30 waterbody sites. Our aim was to determine whether these patterns aligned with the pond-permanence gradient hypothesis, where breeding amphibian abundance is predicted to be highest in ephemeral ponds without large predatory fish. We used Bayesian hierarchical modelling to estimate multi-species abundance from repeated survey (count) data collected over one breeding season.We detected the eggs and larvae of four amphibian species. The best model of abundance included covariates describing two principal component axes associated with waterbody hydrology and landscape composition within a 500-m radius of a site. There was a positive relationship between mean community abundance at a site and hydrological disconnection from the main river channel; however, the common toad (Bufo bufo) was associated with hydrologically connected waterbodies. There was a positive relationship between mean community abundance and a high proportion of forest cover and low cover of agricultural land within a 500-m radius around a site, although this relationship was clear for only two species. There was no support for models containing the number of large predatory fish species detected at a site.Although our results showed that amphibian abundance declined with hydrological connectivity, based on model selection we could not ascribe this relationship to an increased number of large predatory fish species detected in waterbodies close to the main river channel. Differences in life history and habitat requirements are likely to have explained interspecific responses to hydrological connectivity. Our results underscore the importance of addressing amphibian abundance at multiple spatial scales in floodplain wetlands, as landscape composition partly explained patterns in abundance.Application of multi-species abundance modelling allowed us to investigate environmental relationships for common and infrequently detected species. Habitat restoration programmes in floodplains should provide waterbodies disconnected from main river channels as potential amphibian breeding sites and protect or restore forest as terrestrial habitat.

Effect of 80% ethanol or 10% formalin fixation, freezing at -20 °C and staining on Myxobolus (Myxosporea) spores to be deposited in parasitological collections.

The preparation of myxosporeans for the description of myxospores and their preservation as type material in parasitological collections show great variations. Most frequently, formalin and ethanol are used for fixation and Giemsa solution for staining spores. In this work, authors studied the effect of 80% ethanol and 10% formalin fixation, freezing at -20 °C and staining on the size and transparency of two Myxobolus species of cyprinid fishes, M. bramae and M. bliccae spore, and recommended a new method for the deposition of type material to parasitological collections in museums. The studies have commended that fresh spores from mature plasmodia are the best material for measuring the size and studying the inner structures, the number of polar tubules in polar capsules and the morphological characters of the intercapsular appendix. The obtained quantitative data suggest that cryo- and chemical preservation do not have a notable negative effect on spores compared to fresh samples but they decrease the transparency of spores. Staining the spores with Ziehl-Neelsen has proved to be a useful method for studying the fine structure without size reduction, while Giemsa staining induced a shrinkage of spores so it seems to be not ideal for description of a new species. When treating spores of Myxobolus spp. with Lugol's solution, iodinophilous vacuoles in the sporoplasm were not recognised but visualisation of the coils of polar tubules was enhanced. As a type material for newly described species, authors suggest phototypes and spores fixed in 80% ethanol to be deposited into collections, as this preservation method is suitable for subsequent research, such as re-measurements and molecular analysis.

Morphological and molecular studies on two myxosporean infections of cyprinid fishes: Thelohanellus pyriformis from tench and Thelohanellus cf. fuhrmanni from nase.

During surveys on myxosporean parasites of Lake Balaton and River Danube fishes, two Thelohanellus spp. were found on tench (Tinca tinca) and on common nase (Chondrostoma nasus). They were identified as Thelohanellus pyriformis and Thelohanellus cf. fuhrmanni, respectively. Myxospores of T. pyriformis from tench were collected from arteria branchialis afferens of gill filaments. The mature myxospores of this species were pyriform in shape and 19 ± 0.6l (18-19.5) long, 8.2 ± 0.54 (7.5-9) wide, 7.3 ± 0.25 (7-7.5) thick containing polar tubules with 9-10 turns. The plasmodia of Thelohanellus cf. fuhrmanni were collected from under the skin of snout of the common nase. The myxospores were pyriform, 16.3 ± 0.39l (15.5-16.5) long, 6.5 ± 0.55 (6.3-7) wide, 6.3 ± 0.53 (5.8-7) thick containing polar tubules with 6 turns. Small subunit ribosomal DNA sequences of both Thelohanellus species differed from other known myxozoans. The myxospores morphology, histopathology and ssrDNA sequences supported a diagnosis of T. pyriformis from tench and T. cf. fuhrmanni from common nase.

The mixed phylogenetic origin of northern pike (Esox lucius Linnaeus 1758) populations in the Middle Danubian drainage.

BACKGROUND: Pikes, members of genus Esox, are widespread freshwater predators of the northern hemisphere, and important sport fish also. From the Carpathian basin only one species, the northern pike (E. lucius) is noted. At the same time the pike stocks living in this area show high level of phenotypic variance (e.g. various body pattern) and its growth varies highly both among and within populations. These features usually explained by the environmental diversity of the area. Whereas we think that genetic reasons -e.g. the appearance of other/new pike species in the area- may also be responsible for these observed features. Since as no detailed information have been published from the pike populations of this area, so far; we conducted phylogenetic and morphological assay on 88 pike specimens, collected from 49 Middle Danubian sampling sites. RESULTS: Our phylogenetic surveys showed that the northern pike appear in the study area solely, but all the three of its major lineages (Northern, Circumpolar, Southern) were indicated. Only six specimens represent the Northern lineage, collected from the western part of the study area. The Circumpolar and Southern lineages were common in the Carpathian basin, but the Southern lineage showed higher levels of haplotype diversity than the Circumpolar clade. Which indicates that only the Southern lineage is native in the area, while the other two groups could have appeared in the Middle Danubian system either spontaneously or by human introduction. Moreover, the different clades appeared in the same populations, suggesting the opportunity of inter-lineage hybridisation. From the studied morphologicalal features, the number of scales on the lateral line and the head length showed significant differences among the lineages. At the same time the body pattern of the studied individuals seems to be rather influenced by the ontogenic changes than phylogeny. CONCLUSIONS: The high phenotypic variability of Middle Danubian northern pike populations may be due that all of its three major clades appeared and came in secondary contact in the area. In the within watershed spread of the non-native lineages the human stocking/transfer may play a considerable role.

Only one can remain? Environmental and spatial factors influencing habitat partitioning among invasive and native crayfishes in the Pannonian Ecoregion (Hungary).

Biological invasions have increasingly threatened indigenous species, influence metacommunity organisation and consequently, global biodiversity. World-wide expansion of non-indigenous crayfish (NICS) is associated with dramatic changes in species poor indigenous crayfish (ICS) assemblages challenging conservation planning. We analysed long-term changes of crayfish occurrences from the pre-invasion state, through the first appearance of NICS, to their intensive spread in Hungarian waters. Further, we analysed present-day crayfish metacommunity patterns for co-occurrences and influence of spatial and environmental factors. Historic data revealed a marked pre-invasion decline in indigenous noble crayfish Astacus astacus and stone crayfish Austropotamobius torrentium populations, but not in the narrow-clawed crayfish Pontastacus leptodactylus. Historic data provided no direct evidence for the impact of NICS on ICS, rather it supported that NICS often entered areas where ICS had been extinct or were not present at all. Crayfish species extremely rarely co-occurred which could indicate their strong competition and be related to utilization of empty sites by NICS. Crayfish metacommunities were predominantly spatially structured indicating the primary influence of ongoing invasion. Crayfish species also exhibited different environmental preferences mainly along the altitude and temperature gradients. We conclude that the invasion is still in the expanding phase and without an effective conservational program the future of ICS is doubtful in Hungary. Conservation policy should focus on the preservation and reintroduction of the stone and noble crayfishes in highland refugees. Expansion of NICS should be prevented in refugee areas by utilizing possibilities provided by natural and artificial barriers, and education and strict ban should be simultaneously applied to prevent further illegal releases by aquarists.

Multiple stressor effects on alpha, beta and zeta diversity of riverine fish.

We examined the effects of regional scale land use and local scale environmental and biotic stressors on alpha, beta and zeta diversities of native fish communities in wadeable streams and non-wadeable rivers in the Danube basin, Hungary. Relationships among land use and local scale environmental and biotic stressors were weak both in streams and rivers, suggesting that these stressors act relatively independently. Alpha diversity decreased strongly with increasing local scale environmental stressor intensity in rivers. On the contrary, its response to stressors was more obscure in streams, where the best-fit statistical model indicated the importance of the interaction between land use, local scale environmental and biotic stressors, while the secondly ranked model highlighted the negative impact of local scale environmental stressors. Analysis of variance using distance matrices provided evidence that stressors alone and in interactions explained compositional differences of pairs of study sites (beta diversity). Considering the degree of overall degradation, both local (alpha) and among-site (beta and zeta) diversity indices responded to increasing stressor intensity, generally negatively. Riverine fish communities showed higher degrees of similarity (lower beta and higher zeta) than stream fish communities. They also showed increasing similarity (i.e. homogenization) with increasing overall stressor intensity, unlike stream fish communities, which showed no relationship with overall stressor intensity. Our results suggest that the relationships between land use and local scale environmental and biotic stressors can be complex and so do their effects on biodiversity. While stressor specific indices can provide information on the role of specific stressors in some cases, the examination of overall stressor effects is needed to assess realistically the effects of anthropogenic disturbances on native fish diversity. Diversity indices that quantify among-site changes in species composition, such as measures of beta and zeta diversity, can be fruitful for better understanding the role of multiple stressors in structuring ecological communities.

Scavenging behaviour and size-dependent carcass consumption of the black bullhead (Ameiurus melas).

This study examined the size-dependent scavenging behaviour of black bullheads Ameiurus melas under laboratory conditions, using common bleak Alburnus alburnus and pumpkinseed Lepomis gibbosus carcasses. Video camera observations showed that the activity of A. melas was higher at night, but substantial daytime activity was also recorded. Larger A. melas were more active than their smaller conspecifics, especially at night. All size classes exhibited a well-defined sequence of consuming different parts of the carcasses independent of size, but larger individuals tended to consume carcasses more efficiently. Carcasses of the softer-bodied A. alburnus were consumed more readily than those of the bonier L. gibbosus, independent of size. This scavenging behaviour of A. melas might facilitate the invasion success of the species.

Phylogenetic evidence for a new species of Barbus in the Danube River basin.

Three species of small-sized rheophilic Barbus fishes are endemic to and widely distributed throughout the mountain regions in the Danube River basin. In Hungary, barbels referred to as B. petenyi occur in streams in the foothills of the Carpathians near the borders with Slovakia, Ukraine and Romania. However, up to now, no genetic investigations were carried out on rheophilic barbels in this region. This study aims to clarify the taxonomic identity and distribution of the rheophilic barbels in the Hungarian plain based on molecular and morphological analyses. Two mitochondrial genes (cytochrome b, ATPase 6/8) and one nuclear gene (beta-actin intron 2) were sequenced and several morphometric and meristic characters were recorded. Phylogenetic and morphological analyses revealed that there are four genetically distinct lineages among the rheophilic barbels in the Carpathian Basin. The results demonstrated that North-Hungarian Barbus populations belong to B. carpathicus and that B. petenyi presumably does not occur in Hungary. As expected, B. balcanicus was only recorded in samples from the Balkans analyzed for reference. A distinct species, new to science, was discovered to be present in Sebes-Körös River (Crisul Repede) in eastern Hungary and western Romania and is formally described here as B. biharicus Antal, László, Kotlík - sp. nov.