Rudi Verovnik

The colonization of Europe by the freshwater crustacean Asellus aquaticus (Crustacea: Isopoda) proceeded from ancient refugia and was directed by habitat connectivity.

Recent continental‐scale phylogeographic studies have demonstrated that not all freshwater fauna colonized Europe from the classic Mediterranean peninsular refugia, and that northern or central parts of the continent were occupied before, and remained inhabited throughout the Pleistocene. The colonization history of the ubiquitous aquatic isopod crustacean Asellus aquaticus was assessed using mitochondrial COI and a variable part of nuclear 28S rDNA sequences. Phylogeographic analysis of the former suggested that dispersion proceeded possibly during late Miocene from the western part of the Pannonian basin. Several areas colonized from here have served as secondary refugia and/or origins of dispersion, well before the beginning of the Pleistocene. Postglacial large‐scale range expansion was coupled with numerous separate local dispersions from different refugial areas. Connectivity of the freshwater habitat has played an important role in shaping the current distribution of genetic diversity, which was highest in large rivers. The importance of hydrographic connections for the maintenance of genetic contact was underscored by a discordant pattern of mtDNA and nuclear rDNA differentiation. Individuals from all over Europe, differing in their mtDNA to a level normally found between species or even genera (maximal within population nucleotide divergence reached 0.16 ± 0.018), shared the same 28S rRNA gene sequence. Only populations from hydrographically isolated karst water systems in the northwestern Dinaric Karst had distinct 28S sequences. Here isolation seemed to be strong enough to prevent homogenization of the rRNA gene family, whereas across the rest of Europe genetic contact was sufficient for concerted evolution to act.

Phylogeography of subterranean and surface populations of water lice Asellus aquaticus (Crustacea: Isopoda).

The water louse Asellus aquaticus is a widespread, euryoecious species, mostly uniform throughout its range. However, six subspecies are known from the Dinaric karst in the northwestern Balkans. They include some specialized subterranean populations. The pattern of genetic variation among subterranean and surface populations in this hydrographically highly fragmented karst region was investigated using a 653 bp fragment of the mitochondrial gene (COI). Sequencing of 168 individuals from 25 localities revealed 72 haplotypes. amova and methods of phylogenetic reconstruction all uncovered hydrographic structuring of genetic variation of the populations. Nested clade analysis pointed out several fragmentation events, along with some range expansions within hydrographical systems. By superimposing the subterranean mode of life on the phylogeographical pattern, three independent cave colonizations could be inferred within a distance of < 100 km. Caves were invaded after the ancestral surface populations became isolated through vicariant fragmentation. A possible scenario of hydrographic history of the region was constructed combining the molecular data with palaeogeographical information.

Genetic differentiation between two species of the medicinal leech, Hirudo medicinalis and the neglected H. verbana, based on random-amplified polymorphic DNA

Abstract The medicinal leech is one of the few parasitic invertebrates widely used in medicine and as a scientific model object. Because of a dramatic decline in its natural populations, it is subject to considerable conservation effort. Despite all attention, there is confusion regarding the taxonomic status of different morphological forms. The prevailing view is that all varieties of medicinal leech in Europe represent the same species, Hirudo medicinalis. However, the present study based on RAPD molecular markers demonstrates that a second European taxon, H. verbana, forms a distinct species. Phenetic clustering and principal coordinate analysis of eight populations revealed the same basic structure, reflecting taxonomic rather than geographic subdivision. Variation between species explained 60% of the total molecular variance (ΦCT=0.60, P<0.001). Both taxa displayed a significant number of specific RAPD markers. Conversely, no specific fragment supporting the geographic association of both taxa was found. Since the stronghold of commercially exploited medicinal leech populations in southeastern Europe and Turkey belongs to H. verbana, most medicinal and scientific applications probably use this species, not H. medicinalis. Appropriate taxonomic correction of international conservation conventions and legislation is essential.

Random amplified polymorphic DNA diversity among surface and subterranean populations of Asellus aquaticus (Crustacea: Isopoda)

The ecological and evolutionary processes leading to isolation and adaptation of cave animals compared to their surface ancestors are not yet unequivocally understood. In this study the genetic relations of four cave and three surface population of the freshwater crustacean Asellus aquaticus in the Karst region of SW Slovenia and NE Italy were assessed using RAPDs as genetic markers. The results suggest that specialized populations from two caves invaded their subterranean habitat independently, and that their morphological similarity is a result of convergent evolution. Another, less specialized cave population seems to originate from a later colonization of a cave system already inhabited by a more specialized population, but the two populations do not interbreed. This series of temporally and spatially independent invasions has generated a diversity hotspot of non-interbreeding populations of a ubiquitous freshwatercrustacean, which is uniform over most of its range. Genetic variability estimated by the percentage of polymorphic RAPD fragments was similar (86–91%) in most cave and surface populations. Substantially lower values (as low as 49%) were found in two cave populations affected by heavy pollution. Two a priori groupings of populations, traditional subspecies and hydrologically connected groups, were rejected as not significant by means of nested analysis of molecular variance (AMOVA). On the other hand, groupings revealed by UPGMA clustering displayed a significant component of among-group variance. An analysis of gene flow between populations using estimated migration rates confirmed these findings.

The European Grassland Butterfly Indicator: 1990–2011

This report presents the European Grassland Butterfly Indicator, based on national Butterfly Monitoring Schemes (BMS) in 19 countries across Europe, most of them in the European Union. The indicator shows that since 1990 till 2011 butterfly populations have declined by almost 50 %, indicating a dramatic loss of grassland biodiversity. This also means the situation has not improved since the first version of the indicator published in 2005. Of the 17 species, 8 have declined in Europe, 2 have remained stable and 1 increased. For six species the trend is uncertain. The main driver behind the decline of grassland butterflies is the change in rural land use: agricultural intensification where the land is relatively flat and easy to cultivate, and abandonment in mountains and wet areas, mainly in eastern and southern Europe. Agricultural intensification leads to uniform, almost sterile grasslands for biodiversity. Grassland butterflies thus mainly survive in traditionally farmed low‑input systems (High Nature Value (HNV) Farmland) as well as nature reserves, and on marginal land such as road verges and amenity areas. (Less)

Parallels between two geographically and ecologically disparate cave invasions by the same species, Asellus aquaticus (Isopoda, Crustacea)

Caves are long‐known examples of evolutionary replications where similar morphologies (troglomorphies) evolve independently as the result of strong natural selection of the extreme environment. Recently, this paradigm has been challenged based on observations that troglomorphies are inconsistent across taxa and different subterranean habitats. We investigated the degree of replicated phenotypic change in two independent cave invasions by the freshwater isopod Asellus aquaticus; the first in a sulphidic aquifer in Romania, the second in a sinking river in the Dinaric Karst in Slovenia. Both ancestral surface populations still live alongside the subterranean ones. Phylogenetic analyses show independence of the two colonization events, and microsatellite analysis shows no evidence of ongoing genetic exchange between surface and subterranean ecomorphs. The overall morphology has changed dramatically at both sites (50 of 62 morphometric traits). The amount of phenotypic change did not reflect differences in genetic diversity between the two ancestral populations. Multivariate analyses revealed divergent evolution in caves, not parallel or convergent as predicted by the current paradigm. Still, 18 traits changed in a parallel fashion, including eye and pigment loss and antennal elongation. These changes might be a consequence of darkness as the only common ecological feature, because Romanian caves are chemoautotrophic and rich in food, whereas Slovenian caves are not. Overall, these results show that morphologically alike surface populations can diverge after invading different subterranean habitats, and that only about one‐third of all changing traits behave as troglomorphies in the traditional sense.

Case 3508 Maculinea Van Eecke, 1915 (Lepidoptera: lycaenidae): proposed precedence over Phengaris Doherty, 1891

Abstract. The purpose of this application, under Article 23.9.3 of the Code, is to conserve the widely used generic name Maculinea Van Eecke, 1915 in its accustomed usage. The name Maculinea Van Eecke, 1915 is threatened by its senior synonym Phengaris Doherty, 1891. It is proposed that Maculinea be given precedence over the other name whenever the two are considered to be synonyms.

Annotated checklist of Albanian butterflies (Lepidoptera, Papilionoidea and Hesperioidea)

Abstract The Republic of Albania has a rich diversity of flora and fauna. However, due to its political isolation, it has never been studied in great depth, and consequently, the existing list of butterfly species is outdated and in need of radical amendment. In addition to our personal data, we have studied the available literature, and can report a total of 196 butterfly species recorded from the country. For some of the species in the list we have given explanations for their inclusion and made other annotations. Doubtful records have been removed from the list, and changes in taxonomy have been updated and discussed separately. The purpose of our paper is to remove confusion and conflict regarding published records. However, the revised checklist should not be considered complete: it represents a starting point for further research.

Asellus Aquaticus: A Model System for Historical Biogeography

Abstract The aquatic isopod Asellus aquaticus independently populated three cave systems in Europe, where it forms troglomorphic populations all described as separate subspecies. Molecular studies confirm that within northwestern part of Dinaric Mts. additional independent invasions of subterranean habitat occurred, resulting in differently developed troglomorphic traits of those populations. The population present in hidrographically isolated subterranean Reka River drainage is most distant both in terms of molecular markers and morphology and has been described as a separate species A. kosswigi. Similarly, the two populations, inhabiting two separate rivers within Planina Cave, show differences in morphology, mtDNA and microsatellites with very few hybrid specimens at the confluence of the rivers, indicating additional speciation in caves. In a broader view A. aquaticus reached central Europe from the east through current Panonian Plain and spread into the subterranean habitats during karstification linked with Dinaric orogenesis approximately 2–4 Mya ago. Several distinct subspecies both from isolated karst poljes and caves were therefore described. It is still not clear whether the formation of cave adapted populations proceeded in a complete isolation from the surface source population or it could have been achieved in sympatry through ecological speciation. These and other similar questions, provide future opportunities to study this exciting model organism.

Revised checklist of the butterflies of Serbia (Lepidoptera: Papilionoidea)

With studies spanning almost two centuries, butterflies are one of the best known insect groups in Serbia. However, there are still several inconsistencies regarding the number and selection of species included in national checklists published in the last decades. In order to overcome the confusing situation we provide a taxonomically up-to-date checklist of the butterflies for the country, including Kosovo, based on a comprehensive survey of the literature, inspection of available museum collections and from intensive field surveys over the last twenty years. Our aim is also to resolve some long standing problems with species potentially occurring in the country. For this purpose genitalia dissections and DNA barcoding have been used for identification where necessary. The annotated checklist includes 199 species of which Carcharodus orientalis is new for the country and the recently discovered white Anthocharis damone is listed for the first time. We also provide conclusive evidence of the presence of Melitaea ornata in Serbia. Among species listed in the previous species lists we excluded eight species due to inconclusive evidence, and provide a detailed explanation for their exclusion. We hope this publication will stimulate further studies of this important bioindicator group of insects and provide the basis for their conservation in the country.