Odonata Assemblages in Urban Semi-Natural Wetlands.

As the human population in urban areas is continuously growing, urbanization is one of the greatest threats to biodiversity. To mitigate the negative effects, the inclusion of blue zones (aquatic habitats) in modern urban development practices is strongly recommended, as they could be beneficial for the local biodiversity conservation. Odonata are a flagship group and are widely used in freshwater conservation as ecological indicators of habitat integrity and health. However, our understanding of their ecological requirements in urban landscapes is not yet complete. Therefore, we analyzed the taxonomic and functional diversity of Odonata in a semi-natural wetland in the Croatian capital. This study was conducted in the summers of 2020 and 2023. Most taxonomic and functional assemblage metrics were comparable between the two main habitat types, anthropogenically disturbed and natural oxbow lakes. However, significant differences were found in relation to the time scale, where most metrics were lower in 2023, indicating the negative impact of extreme climate events (including droughts) that occurred in this region after 2020. With 19 species recorded, our results indicate that semi-natural urban wetlands, especially natural oxbow lakes, have great potential to function as good habitats for Odonata, where even some species of conservation concern were detected. When developing landscape management plans in urban areas, it is essential to consider the importance of habitat heterogeneity in terms of good structure of aquatic macrophytes (presence of submerged, emergent and floating vegetation), which would ensure the most suitable habitat conditions for local Odonata species.

A collaborative backbone resource for comparative studies of subterranean evolution: The World Asellidae database.

Transition to novel environments, such as groundwater colonization by surface organisms, provides an excellent research ground to study phenotypic evolution. However, interspecific comparative studies on evolution to groundwater life are few because of the challenge in assembling large ecological and molecular resources for species-rich taxa comprised of surface and subterranean species. Here, we make available to the scientific community an operational set of working tools and resources for the Asellidae, a family of freshwater isopods containing hundreds of surface and subterranean species. First, we release the World Asellidae database (WAD) and its web application, a sustainable and FAIR solution to producing and sharing data and biological material. WAD provides access to thousands of species occurrences, specimens, DNA extracts and DNA sequences with rich metadata ensuring full scientific traceability. Second, we perform a large-scale dated phylogenetic reconstruction of Asellidae to support phylogenetic comparative analyses. Of 424 terminal branches, we identify 34 pairs of surface and subterranean species representing independent replicates of the transition from surface water to groundwater. Third, we exemplify the usefulness of WAD for documenting phenotypic shifts associated with colonization of subterranean habitats. We provide the first phylogenetically controlled evidence that body size of males decreases relative to that of females upon groundwater colonization, suggesting competition for rare receptive females selects for smaller, more agile males in groundwater. By making these tools and resources widely accessible, we open up new opportunities for exploring how phenotypic traits evolve in response to changes in selective pressures and trade-offs during groundwater colonization.

Metal bioaccumulation in stygophilous amphipod Synurella ambulans in the hyporheic zone: The influence of environmental factors.

Given the increasing need to protect vulnerable freshwater ecosystems and make them more resilient to human use and climate change, biomonitoring of the hyporheic zone (HZ), which plays a critical role in pollution attenuation, is essential. The aim of the present study was to assess the potential of the amphipod species Synurella ambulans as a bioindicator of metal contamination in the HZ of the Sava River (Croatia). Amphipods were collected during the four seasons at two sampling sites (average sampling depth 55 cm) differing in type (agricultural and urban) and intensity (diffuse and point source contamination) of anthropogenic influence, one located upstream (Medsave), and the other downstream (Jarun) of the wastewater treatment plant discharge. Concentrations of Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Rb, Sn, Zn, Ca, K, Mg and Na were measured in the interstitial water, sediments and specimens of S. ambulans by HR ICP-MS. Physicochemical parameters (temperature, DO, O2 saturation, pH, conductivity, alkalinity, total water hardness, CODKMnO4, nutrients) were measured in the interstitial water, while organic carbon was measured in the interstitial water and sediments. Metal concentrations in interstitial water and sediments were below thresholds set by environmental quality standards. Metal concentrations in S. ambulans were classified as follows: higher at the Jarun site (Al, Cr, Fe, Ni, Pb, Sn), higher at the Medsave site (Cd, Cu, Rb) and mostly comparable at both sites (Co, Mn, Zn). Bioaccumulation factors were generally higher at Jarun, with average values ranging from 322 to 143,278 L kg-1. Bioaccumulation of metals in S. ambulans depended on various environmental factors, with metal exposure level and dissolved macro elements showing the strongest association with metals accumulated in S. ambulans. The findings provided the first evidence on the suitability of S. ambulans as a good bioindicator of chronic metal contamination in the HZ.

Taxonomy of Alpioniscus (Illyrionethes): A. magnus and three new species from the Dinaric Karst (Isopoda: Oniscidea: Trichoniscidae).

Alpioniscus (Illyrionethes) is a widespread subgenus of Trichoniscidae in the Dinaric Karst with 15 nominal troglobiotic species. Here we redescribe Alpioniscus (Illyrionethes) magnus and describe three new species (A. lossinii n. sp., A. drazinai n. sp., and A. mandalinae n. sp.) restricted to coastal areas. As a result, 18 nominal Illyrionethes species are known from the Dinaric Karst.

Is the spatial distribution of lentic water mite assemblages (Acari: Hydrachnidia) governed by prey availability?

Predation is a biotic interaction that links water mites to different taxonomic groups of benthic invertebrates. Diptera larvae (Chironomidae) and microcrustaceans (Cladocera, Copepoda, Ostracoda) are known to be the most commonly preferred prey by water mites. Although these predatory interactions are known from observations and ex situ investigations, the distribution patterns and co-occurrence of water mites and their prey in littoral lentic habitats have been poorly studied. Our goal was to determine whether predation might serve as a significant factor in water mite assemblage composition and distribution. Samples were taken from littoral zones of 21 artificial lakes (reservoirs) in the Dinaric Western Balkan ecoregion of Croatia. At every site, 10 samples were collected with regard to shore slope in depth zones of up to 1 m. In total 490 samples were collected from April 2016 to July 2017. Data analysis showed that the spatial distribution of water mites was partially correlated with the environmental parameters we measured (explaining 45.1% of total water mite variation). A positive correlation between water mite abundance and diversity and depth gradient (favouring more stable conditions in deeper parts of the littoral zone) was also observed. Finally, water mite abundance and species richness variation were found to be best predicted by the abundance of potential prey groups. Predation was statistically determined to be a strong dispersion variable, most probably influencing both spatial distribution and composition of water mite assemblages.

Water mite (Acari: Hydrachnidia) diversity and distribution in undisturbed Dinaric karst springs.

Large undisturbed karst springs in the Dinaric region are particularly interesting and extremely valuable and rare ecosystems, housing diverse and yet undiscovered water mite species. The complex life history traits of water mites include several life stages that are interconnected with other freshwater arthropod groups, making them important components of freshwater fauna. This study examined whether the differences in water mite assemblages between Mediterranean and alpine karst springs were driven by seasonal and/or regional changes in environmental conditions. The first ecological data on water mite dynamics and distribution in the Dinaric region of Croatia are presented. Water mites were represented by 11 taxa, decreasing in taxon richness from alpine to Mediterranean springs. Spatial patterns of water mite assemblages at the regional scale were best explained by water temperature, dissolved oxygen and water velocity. Pseudotorrenticola rhynchota and Oxus setosus were exclusively found in springs with higher water temperature and velocity, whereas the genera Aturus sp. and Woolastookia rotundifrons were exclusively recorded in the alpine springs, where oxygen concentrations were significantly higher. Atractides sp., Lebertia sp. and Sperchon sp. were recorded in most springs. Water mite abundances had a summer peak in the Mediterranean region, whereas two smaller peaks were observed in the spring and fall in the alpine region. Regional differences in the seasonality of water mite abundance were most likely patterned by the emergence of insect crenofauna.

Benthos-drift relationships as proxies for the detection of the most suitable bioindicator taxa in flowing waters - a pilot-study within a Mediterranean karst river.

Mediterranean karst aquifers are sensitive systems vulnerable to contamination, exhibiting high rates of diversity and endemicity. In the present pilot-study, we aimed to detect the most suitable bioindicators of contaminant accumulation and mobilization within a Mediterranean karst river (Krka River, Croatia), whose lowermost sections belong to a designated protection area (national park). To meet our goal, we sampled water, drift and benthos (macroinvertebrates and periphytic microfauna) at the two Krka River sites, located upstream and downstream from town Knin and its urban influences. We compared: 1) environmental conditions (water physico-chemical parameters, trace- and macro-element concentrations); 2) abundance and diversity of periphyton and macroinvertebrate taxa constituting benthos; and 3) macroinvertebrate benthos-drift relationships between the two sites. Despite higher values of all measured physico-chemical parameters, and most trace- and macro-element concentrations at the urban-influenced site, the concentrations of contamination indicators (i.e., COD, nutrients, metals) at both sites were generally low. This is likely a result of specific "self-purification ability" of the Krka River, mediated by relatively high contaminant retention potential of the underlying tufa (i.e., calcareous) and/or macrophyte substrates. Between-site differences in water quality further affected the spatial variation of macrozoobenthos, drift, and periphytic microfauna. We suggest that increased COD and orthophosphate concentration, and macrophyte presence at the urban-influenced site, supported higher densities and diversity of benthic organisms dominated by eurivalent (i.e., contamination-tolerant) taxa. The most numerous macroinvertebrate taxa in benthos were amphipod Gammarus balcanicus and the representatives of the endemic Dinaric karst taxa - gastropods Emmericia patula and Radomaniola curta germari, and another amphipod Echinogammarus acarinatus. Although we expected to observe significantly increased drift at the urban-influenced site due to the degraded environmental conditions, it was not observed. The observed benthos-drift patterns suggest that freshwater amphipods (i.e., gammarids), which were found most numerous in drift, could be considered as the most suitable bioindicators of a contaminant (i.e., metal) accumulation and mobilization within karst aquifers comparable to Krka River.

Spatial distribution and seasonal changes of mayflies (Insecta, Ephemeroptera) in a Western Balkan peat bog.

Peat bogs are unique wetland ecosystems of high conservation value all over the world, yet data on the macroinvertebrates (including mayfly assemblages) in these habitats are still scarce. Over the course of one growing season, mayfly assemblages were sampled each month, along with other macroinvertebrates, in the largest and oldest Croatian peat bog and an adjacent stream. In total, ten mayfly species were recorded: two species in low abundance in the peat bog, and nine species in significantly higher abundance in the stream. Low species richness and abundance in the peat bog were most likely related to the harsh environmental conditions and mayfly habitat preferences. In comparison, due to the more favourable habitat conditions, higher species richness and abundance were observed in the nearby stream. Three of the recorded species, Caenis luctuosa from the peat bog, and Eurylophella karelica and Leptophlebia marginata from the stream are new records for the Croatian mayfly fauna. Typical Central European life cycle patterns were confirmed for several species (e.g. Baetis vernus, Nigrobaetis niger, Electrogena ujhelyii), while for several others (e.g. Habrophlebia fusca, Paraleptophlebia submarginata) some discrepancies were observed. Therefore, these results provide new and valuable information on the ecology of mayflies in peat bog habitats.

The limits of cryptic diversity in groundwater: phylogeography of the cave shrimp Troglocaris anophthalmus (Crustacea: Decapoda: Atyidae).

Recent studies have revealed high local diversity and endemism in groundwaters, and showed that species with large ranges are extremely rare. One of such species is the cave shrimp Troglocaris anophthalmus from the Dinaric Karst on the western Balkan Peninsula, apparently uniform across a range of more than 500 kilometers. As such it contradicts the paradigm that subterranean organisms form localized, long-term stable populations that cannot disperse over long distances. We tested it for possible cryptic diversity and/or unexpected evolutionary processes, analysing mitochondrial (COI, 16S rRNA) and nuclear (ITS2) genes of 232 specimens from the entire range. The results of an array of phylogeographical procedures congruently suggested that the picture of a widespread, continuously distributed and homogenous T. anophthalmus was wrong. The taxon is composed of four or possibly five monophyletic, geographically defined phylogroups that meet several species delimitation criteria, two of them showing evidence of biological reproductive isolation in sympatry. COI genetic distances between phylogroups turned out to be a poor predictor, as they were much lower than the sometimes suggested crustacean threshold value of 0.16 substitutions per site. Most results confirmed the nondispersal hypothesis of subterranean fauna, but the southern Adriatic phylogroup displayed a paradoxical pattern of recent dispersal across 300 kilometers of hydrographically fragmented karst terrain. We suggest a model of migration under extreme water-level conditions, when flooded poljes could act as stepping-stones. In the north of the range (Slovenia), the results confirmed the existence of a zone of unique biogeographical conflict, where surface fauna is concordant with the current watershed, and subterranean fauna is not.