The Rhyacophila fasciata Species Complex (Trichoptera: Rhyacophilidae) in Central Europe with description of a new species, Rhyacophila loeffleri Valladolid & Waringer, n. sp., based on morphological, genetic and ecological evidence.

The morphology of the larva, pupa, male, and female of Rhyacophila loeffleri Valladolid & Waringer, n. sp., from Austria, Czech Republic, France, and Italy is described. Data from molecular analysis are also provided, based on mitochondrial cytochrome c oxidase subunit I (mtCOI), including sequences from German and Swiss specimens, and compared with those of samples of the nominate species R. fasciata Hagen 1859, as well as with other species and subspecies in the Rhyacophila fasciata Species Complex: R. septentrionis McLachlan 1865, R. denticulata McLachlan 1879, R. sociata Navs 1916, R. kykladica Malicky & Sipahiler 1993, R. delici Kuini & Valladolid 2020, R. fasciata viteceki Valladolid & Kuini 2020 and R. macedonica Karaouzas, Valladolid & Ibrahimi 2022. Our study revealed morphological differences between R. fasciata and R. loeffleri, as well as genetic differences among the other taxa of the R. fasciata Complex, supporting the recognition of R. loeffleri as a new species. Also, Rhyacophila fasciata viteceki is recognized as a distinct species, R. viteceki (status promotus), based on results of the molecular analysis.

The Rhyacophila fasciata Group in Europe: Rhyacophila macedonica Karaouzas, Valladolid amp; Ibrahimi (n. sp.) from Greece, Kosovo, Republic of North Macedonia and Serbia (Trichoptera: Rhyacophilidae).

The morphology of all postembryonic stages (larva, pupa, male, and female) of Rhyacophila macedonica Karaouzas, Valladolid, Ibrahimi n. sp. from Greece, Republic of North Macedonia, Kosovo and Serbia was examined. Morphological data were supplemented by a molecular analysis of the mitochondrial cytochrome C oxidase subunit I (COI) and compared with samples of the nominate species Rhyacophila fasciata Hagen 1859, as well as with other species and subspecies in this group: Rhyacophila septentrionis McLachlan 1865, Rhyacophila denticulata McLachlan 1879, Rhyacophila sociata Navs 1916, Rhyacophila kykladica Malicky Sipahiler 1993, Rhyacophila fasciata delici Kucinic Valladolid 2020, and Rhyacophila fasciata viteceki Valladolid Kucinic 2020. Our results revealed morphological differences between the nominate species and R. macedonica, as well as genetic differences among the taxa in the Rhyacophila fasciata Group, so we propose R. macedonica as a new species of the Group. Based on the new molecular data, we also elevate R. f. delici to the status of a distinct species, R. delici (status promotus).

Would delineation of nitrate vulnerable zones be improved by introducing a new parameter representing the risk associated with soil permeability in the Land Use-Intrinsic Vulnerability Procedure?

Most methods for mapping groundwater vulnerability are based on the excessively simplistic approach that aquifer recharge is produced by vertical infiltration. The novel Land Use-Intrinsic Vulnerability (LU-IV) procedure assesses groundwater vulnerability to nitrate pollution over the entire territory, including aquifers catchment areas. In this research, it was analysed if the delineation of nitrate vulnerable zones (NVZs) would be improved by introducing a new parameter representing the risk associated with soil permeability (parameter S) in the procedure. Different versions of parameter S were tested: S_HC (risk associated with soil hydraulic conductivity), S_St+G+S (risk associated with the stone, gravel and sand fraction of the soil) and S_C (risk associated with the clay fraction). The study was undertaken in the catchment areas of the Oja and Tirón alluvial aquifers (Spain). The efficacy of the following six models was compared: Model 1 (original LU-IV procedure), Model 2 (LU-IV' procedure using parameter S_HC), Model 3 (LU-IV' procedure using parameter S_St+G+S), Model 4 (LU-IV' procedure using parameter S_C), Model 5 (LU-DRASTIC-COP procedure, based on DRASTIC-COP method), and Model 6 (designated NVZ). Catchment scale validations of the six models showed similar, highly significant correlations between the percent coverages of the estimated NVZs and those of the alluvial areas polluted by nitrate for Models 1 to 4. Models 5 and 6 did not show any significant results. In light of these results, Models 1 to 4 were considered the best predictors of nitrate pollution and the best methods for NVZ delineation. Results support the idea that including a parameter S in the LU-IV' procedure is not essential since equivalent results were obtained from the original LU-IV procedure. So, the LU-IV procedure should be considered the best and simplest method of those tested for accurately delineating NVZs.

N and P behaviour in alluvial aquifers and in the soil solution of their catchment areas: How land use and the physical environment contribute to diffuse pollution.

The role of land use and the physical environment in N and P pollution of alluvial aquifers was analysed at three levels of information: (1) aquifer (N and P in groundwater), (2) soil transect (potentially leachable N and P in the soil solution) and (3) aquifer's catchment area. The study was carried out in the Oja and Tirón alluvial aquifers and their catchment areas (northern Spain). Nitrate was the dominant N form, both in groundwater and the soil solution of aquifers' catchment areas. Orthophosphate and organic-P were the codominant P forms in the aquifers. Orthophosphate was the main form in the soil solution. During the period 2005-2017 no significant decrease in nitrate pollution was observed, suggesting the need to review current Nitrate Vulnerable Zone (NVZ) designations. Since nitrate is highly mobile, it tended to accumulate in stagnation zones at the lower reaches of the aquifers. P did not accumulate in the same zones due to its low solubility. Principal component analyses (PCAs) of the aquifers, soil transects and aquifers' catchment areas revealed that the observation scale influences the environmental factors that can be detected as intervening in groundwater pollution. At the aquifer scale, links were found between nitrates and land use, topographic, hydrogeological and climatic factors. The protective effect of natural areas against nitrate pollution was noteworthy, while agriculture was associated with pollution. At the soil transect scale, an altitudinal gradient governed soil particle size distribution and land use, separating mountain forest soils from agricultural soils. The negative relationship between clay contents vs. nitrate and orthophosphate in the soil solution pointed to a regulatory role of clay. At the catchment scale, the size and physical characteristics of the catchments and land use distribution determined macronutrient availability in the soil solution and, in turn, N and P groundwater distribution.

The Rhyacophila fasciata Group in Europe: Rhyacophila fasciata Hagen 1859 and formerly synonymized species (Trichoptera: Rhyacophilidae), with new description of Rhyacophila fasciata and Rhyacophila septentrionis McLachlan 1865 (stat. prom.).

The presence and distribution of Rhyacophila fasciata Hagen 1859 in Europe were revised, based on bibliographic study, collection specimens, and new material collected in different countries. The status of formerly synonymized species, Rhyacophila ferruginea (Scopoli 1763) and Rhyacophila septentrionis McLachlan 1865 was also assessed. The type of R. ferruginea is missing, the taxon is still unidentified, and thus we propose Rhyacophila ferruginea as a nomen dubium. Morphological features and genetic evidence revealed that R. septentrionis differs from R. fasciata, so we propose to change its status to status resurrectus. We therefore include new descriptions of the different stages (larva, pupa, male, and female) of R. fasciata and of R. septentrionis, together with a molecular analysis based on mitochondrial cytochrome oxidase I (mtCOI) and ecological notes. The species R. coppai Oláh 2020 NEW SYNONYM and R. soreda Coppa Oláh 2020 NEW SYNONYM are synonyms of R. sociata Navás 1916; the species R. kopasa Oláh Coppa 2020 NEW SYNONYM and R. rova Oláh Coppa 2020 NEW SYNONYM are synonyms of R. denticulata McLachlan 1879; the species R. matrensis Oláh Szczesny 2020 is probably a synonym of R. fasciata, so more study of this species is needed in order to confirm or deny that it is a valid species.

The Rhyacophila fasciata Group in Croatia and Bosnia and Herzegovina: Rhyacophila f. fasciata Hagen 1859 and the description of two new subspecies, Rhyacophila fasciata delici Kucinic Valladolid (ssp. nov.) from Croatia and Bosnia and Herzegovina and Rhyacophila fasciata viteceki Valladolid Kucinic (ssp. nov.) from Bosnia and Herzegovina (Trichoptera: Rhyacophilidae).

We present the description of two new subspecies of the Rhyacophila fasciata Group: Rhyacophila fasciata delici Kucinic Valladolid (ssp. nov.), broadly distributed in Croatia and present also in Bosnia and Herzegovina, and R. fasciata viteceki Valladolid Kucinic (ssp. nov.), found in Bosnia and Herzegovina. Our study of the morphology of adults, as well as our analysis of the barcode region of the mitochondrial cytochrome oxidase I (mtCOI) gene and geographical distribution confirm the differences of the two new subspecies with the nominal species R. f. fasciata, also found in both countries.

The Rhyacophila fasciata Group in Greece: Rhyacophila kykladica Malicky amp; Sipahiler 1993 (stat. prom.) (Trichoptera: Rhyacophilidae). Morphological description, genetic and ecological features.

The morphology of the different stages (larva, pupa, male, and female) of Rhyacophila fasciata kykladica Malicky Sipahiler 1993 was examined as a basis for the description of the taxon. Morphological data were supplemented by molecular analysis of mitochondrial cytochrome C oxidase subunit I (mtCOI), which were compared with samples of the nominate subspecies Rhyacophila fasciata fasciata Hagen 1859, as well as with other new species in this group, Rhyacophila denticulata McLachlan 1879 and Rhyacophila sociata Navás 1916. Our results revealed genetic differences between all the taxa; therefore, a change of taxonomic status of R. fasciata kykladica to R. kykladica (stat. prom.) is proposed.

Assessment of the risks of N-loss to groundwater from data on N-balance surplus in Spanish crops: An empirical basis to identify Nitrate Vulnerable Zones.

The aim of this research was to conduct an empirical assessment of the risks of N-loss to groundwater associated with land use (LU), based on annual data on the net N-balance surplus in Spanish crops. These data were used to generate a detailed risk rating system reflecting the potential risks of N-loss from agriculture. The new LU ratings were used to assess the specific vulnerability of groundwater to nitrate pollution, by using the LU-IV procedure (Arauzo 2017). The study area included the catchment areas of 12 alluvial aquifers associated to tributaries of the Ebro River (Spain). Most of the alluvial aquifers were chronically polluted by nitrate, with only a few remaining unaffected by pollution. The LU maps from two different base maps (MCAE 2000-09; SIOSE 2011) were used to generate the respective versions of the map of vulnerability to nitrate pollution using the LU-IV procedure. Potential nitrate vulnerable zones (NVZ) were extracted from different models of vulnerability for comparison with the map of groundwater nitrate content. The models compared were the following: model A (LU-IV procedure, based on MCAE 2000-09 and using LU ratings from N-surpluses in Spanish crops), model B (LU-IV procedure, based on SIOSE 2011 and using LU ratings from N-surpluses in Spanish crops), model C (LU-IV procedure, based on MCAE 2000-09 and using LU ratings from bibliographical references; Arauzo, 2017), model D (IV index), model E (DRASTIC index), and model F (GOD index). Results confirmed, as expected, that models A and B proved to be the best risk predictors, both for polluted groundwater areas and for areas at risk of being polluted. These results support the high level of reliability of the LU-IV procedure, when applying the LU ratings obtained empirically from the N-surpluses.

The Rhyacophila fasciata Group in Western Europe: Confirmation of Rhyacophila denticulata McLachlan 1879 (stat. prom.) and Rhyacophila sociata Navás 1916 (stat. res.), based on morphological and molecular genetic evidence (Trichoptera: Rhyacophilidae).

In order to check the presence and distribution of Rhyacophila fasciata fasciata Hagen 1859 (species described from Austria) and R. fasciata denticulata McLachlan 1879 in the Iberian Peninsula, we studied the morphology of Spanish, French, and Austrian specimens, together with their mitochondrial cytochrome c oxidase (mtCOI). We observed that the individuals considered to date as R. fasciata denticulata are in fact two different species: R. denticulata, presently known from France and possibly in some rivers of the Basque Country (Spain), and R. sociata Navás 1916 distributed in Spain and France. These two species are also different from the reference species (R. fasciata) from Austria, so we propose a change in the taxonomic status of R. fasciata denticulata to R. denticulata (stat. prom.) and the restoration of R. sociata (stat. res.), with the designation of a neotype, due to the loss of the holotype.

Vulnerability of groundwater resources to nitrate pollution: A simple and effective procedure for delimiting Nitrate Vulnerable Zones.

This research was undertaken to further our understanding of the factors involved in nonpoint-source nitrate pollution of groundwater. The shortcomings of some of the most commonly used methods for assessing groundwater vulnerability have been analysed and a new procedure that incorporates key improvements has been proposed. The new approach (LU-IV procedure) allows us to assess and map groundwater vulnerability to nitrate pollution and to accurately delimit the Nitrate Vulnerable Zones. The LU-IV procedure proved more accurate than the most widely used methods to assess groundwater vulnerability (DRASTIC, GOD), when compared with nitrate distribution in the groundwater of 46 aquifers included in the study (using the drainage basin as the unit of analysis). The proposed procedure stands out by meeting the following requirements: (1) it uses readily available parameters that provide enough data to feed the model, (2) it excludes redundant parameters, (3) it avoids the need to assign insufficiently contrasted weights to parameters, (4) it assess the whole catchment area that potentially drains N-polluted waters into the receptor aquifer, (5) it can be implemented within a GIS, and (6) it provides a multi-scale representation. As the LU-IV procedure has been demonstrated to be a reliable tool for delimiting NVZ, it could be particularly interesting to use it in countries where certain types of environmental data are either not available or have only limited availability. Based on this study (and according to the LU-IV procedure), it was concluded that an area of at least 1728km2 should be considered as NVZ. This sharply contrasts with the current 328km2 officially designated in the study area by the Spain's regional administrations. These results highlight the need to redefine the current NVZ designation, which is essential for an appropriate implementation of action programmes designed to restore water quality in line with Directive 91/676/EEC.