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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. [ABSTRACT FROM AUTHOR]

The three most abundant and species-rich genera of erpobdellid leeches - Erpobdella, Dina and Trocheta - have traditionally been distinguished by their annulation pattern. Erpobdella has five unsubdivided annuli per somite, in Dina the last annulus is widened and subdivided once, and in Trocheta, the last and often also the first annulus is further subdivided. The last two genera are often referred to as the subfamily Trochetinae. The presented molecular phylogenetic analysis, based on mitochondrial 12S rDNA sequences and nuclear ITS2 sequences,has revealed a sister relationship between Trocheta and Erpobdella rather than between Trocheta and Dina. Furthermore, the annulation pattern has proved to be inappropriate for the diagnosis of Dina and Trocheta. The taxon Trocheta bykowskii krasense has been classified on the basis of its Trocheta-like annulation; however, according to the molecular phylogeny, it belongs to the Dina clade. A pair of diagnostic PCR primers were developed to identify unambiguously individuals with intermediate annulation. In summary, (1) the traditional genus diagnosis of Dina and Trocheta is no longer tenable, and the genera are not always morphologically distinguishable by traditional characters;(2) T. b. krasense Sket, 1968, should be re-classified as Dina krasensis; (3) the subfamily Trochetinae should be annulled because it does not represent a monophyletic assemblage. [ABSTRACT FROM AUTHOR]

We studied species richness patterns of obligate subterranean (troglobiotic) beetles in the Dinaric karst of the western Balkans, using five grid sizes with cells of 80 × 80, 40 × 40, 20 × 20, 10 × 10, and 5 × 5 km. The same two hotspots could be recognized at all scales, although details differed. Differences in sampling intensity were not sufficient to explain these patterns. Correlations between number of species and number of sampled localities increased with increasing cell size. Additional species are expected to be found in the region, as indicated by jackknife 1, jackknife 2, Chao2, bootstrap, and incidence-based coverage (ICE) species richness estimators. All estimates increased with increasing cell size, except Chao2, with the lowest prediction at the intermediate 20 × 20 km cell size. Jackknife 2 and ICE gave highest estimates and jackknife 1 and bootstrap the lowest. Jackknife 1 and bootstrap estimates changed least with cell size, while the number of single cell species increased. In the highly endemic subterranean fauna with many rare species, bootstrap may be most appropriate to consider. Positive autocorrelation of species numbers was highest at 20 × 20 km scale, so we used this cell size for further analyses. At this scale we added 137 localities with less positional accuracy to 1572 previously considered, and increased 254 troglobiotic species considered to 276. Previously discovered hotspots and their positions did not change, except for a new species-rich cell which appeared in the south-eastern region. There are two centres of troglobiotic species richness in the Dinaric karst. The one in the north-west exhibited high species richness of Trechinae (Carabidae), while in the south-east, the Leptodirinae (Cholevidae) were much more diverse. These centres of species richness should serve as the starting point for establishing a conservation network of important subterranean areas in Dinaric karst. [ABSTRACT FROM AUTHOR]

BelgrandiellaA. J. Wagner, 1927 is a genus comprising minute snails with conic or turriform shells that are facultative stygobionts, inhabiting subterranean waters and springs. Few specimens or even empty shells of this taxon can be found, so the taxonomy is based mostly on the shell morphology and geographic distribution. Anatomy is known for a few taxa, and its usefulness is restricted. This, coupled with a popular idea that isolation is the first factor shaping divergence and speciation of the spring/cave fauna, resulted in dozens of nominal species ofBelgrandiella, known from shells at a given locality. Herein we present shells, anatomy and four molecular markers (nuclear H3, 18S and 28S and mitochondrial cytochrome oxidase subunit I) inBelgrandiellafrom 16 localities in Slovenia, northern Croatia, and Bosnia and Herzegovina. Five sites are type localities ofBoleana umbilicata, Belgrandiella kusceri,Belgrandiella krupensis,Belgrandiella zermanicaandBelgrandiella robusta,respectively. Considering the shell and geographic distribution, we have determinedB. cf.robustaBelgrandiellacf.croatica,Belgrandiellacf.fontinalis,Belgrandiellacf.kuesteri,Belgrandiellacf.pagetiandBelgrandiellacf.koprivnensisThe shells of all, as well as penes and female reproductive organs of two putative, species have been illustrated. Molecular analysis indicates four distinct clades, most probably of species rank forB. kusceri,B. cf.kuesteri,B. cf.fontinalisandB. cf.koprivnensis. Four nominal species (B. krupensis,B. robusta,B. umbilicataandB. zermanica) and one genus (Boleana) must be synonymised withBelgrandiella kusceri, and there are no more than four valid, biological species in the studiedBelgrandiella. The molecular divergence is completely unmirrored by high morphological variability. Hence the level of endemism is markedly overestimated in these snails. [ABSTRACT FROM AUTHOR]

The Vjetrenica cave in the Dinaric Karst hosts a worldwide extraordinarily high cave biodiversity. Beside a diverse and specialized cave fauna, sprout-like formations attached to the bed of the cave stream were observed and described, but not further characterized, almost a century ago. Here we investigated these sprout-like microbial aggregates by the rRNA approach and detailed microscopy. Based on fluorescence in situ hybridization and ultrastructural analysis, the sprout-like formations are morphologically highly organized, and their core consists of a member of a novel deep-branching lineage in the bacterial phylum Nitrospirae. This organism displays an interesting cellular ultrastructure with different kinds of cytoplasmic inclusions and is embedded in a thick extracellular matrix, which contributes to the stability and shape of the aggregates. This novel bacterium has been provisionally classified as ' Candidatus Troglogloea absoloni.' The surface of the sprout-like aggregates is more diverse than the core. It is colonized by a bacterial biofilm consisting primarily of filamentous Betaproteobacteria, whereas other microbial populations present in the crust include members of the Bacteriodetes, Gammaproteobacteria, Actinombacteria, Alphaproteobacteria, and Planctomycetes, which are intermingled with mineral inclusions. This study represents the first thorough molecular and ultrastructural characterization of the elusive sprout-like bacterial aggregates, which are also found in other cave systems of the Dinaric Karst. The discovery of Ca. Troglogloea absoloni contributes to the known biodiversity of subterranean ecosystems and especially of macroscopic structures formed in caves by microorganisms, whose composition and ecological function often remain enigmatic. [ABSTRACT FROM AUTHOR]

Natural history museum collections harbour valuable information on species. The usefulness of such data critically depends on the accurate identification of species, which has been challenged by introduction of molecular techniques into taxonomy. However, most collections may suffer from DNA degradation, due to age and/or improper preservation; hence the identification of specimens depends solely on morphological features. This study explores how and to what extent morphological data can help to solve ambiguous taxonomic cases based on selected species concepts and with the use of operational criteria in a species-hypothesis testing procedure. The studied taxon, the Niphargus tatrensis species complex, comprises freshwater subterranean amphipods, distributed across Central Europe, the taxonomic status of which was debated extensively between 1930 and 1960. Using the general species concept, character- and tree-based operational criteria reveal northern and southern diagnosable and exclusive lineages identified here as N. tatrensis Wrześniowski, 1888 and N. scopicauda sp. n., respectively. The remaining populations represent the non-exclusive N. aggtelekiensis Dudich, 1932, which occurs from the eastern Alps to Hungary. In the entire complex, altitudinal distribution is largely limited to areas above 400 m, where the mean annual temperature never exceeds 9°C. Seemingly well-defined distributional ranges of N. tatrensis and N. aggtelekiensis are fragmented in an ecological sense, which raises the question whether two of the three species recognised here actually consist of several unidentified taxa. Morphological similarity between the species, numerous polymorphic features, and the association with cool temperatures lead to a hypothesis in which fragmentation of the ancestral range occurred during post-Pleistocene climate warming, reducing gene flow across lowland populations due to niche conservatism of the ancestral species and/or to invasion of competitive species along the Danube and Drava rivers. The results are discussed regarding how old museum samples are conducive to more detailed molecular-taxonomic and conservation studies. [ABSTRACT FROM AUTHOR]

Macroaggregates in the northern Adriatic, presently considered as a product of agglomeration of macromolecular dissolved organic matter (DOM) into macrogels and particulate organic matter (POM), are primarily associated with phytoplankton, mostly diatom, blooms occurring in late spring during favorable environmental conditions. Composition of mucous macroaggregates in the northern Adriatic, collected at different evolutionary stages, was studied using spectroscopic (solid-state 13C NMR, FT-IR and 1H NMR), elemental and X-ray (XRD) analyses. The results confirm the general structure of macroaggregates, including four major classes of structural elements: carbohydrates, ester and amide functional groups, aliphatic, and organosilicon components. The water-soluble fraction of macroaggregates is mostly composed of carbohydrates with a minor content of aliphatic components, and is similar to ultrafiltered dissolved organic matter (UDOM) from the cultured diatom Cylindroteca fusiformis, suggesting the importance of diatom exudates as macroaggregate precursors. The aliphatic and organosilicon components prevailed in the water-insoluble fraction. The macroaggregate gel is mainly stabilized by organo-mineral interactions with calcite, quartz and clay minerals. The results also indicate the preservation of an N containing component within the macroaggregate refractory organic matrix. [Copyright &y& Elsevier]

Climate changes have substantial impacts on the geographic distribution of montane lakes and evolutionary dynamics of cold-adapted species. Past climate cooling is hypothesized to have promoted the dispersal of cold-adapted species via montane lakes, while future climate warming is thought to constrain their distributions. We test this hypothesis by using phylogeographic analysis and niche modeling of the Holarctic crustacean Gammarus lacustris with global sampling comprised of 567 sequenced individuals and 3180 occurrence records. We found that the species arose in Tian Shan in Central Asia and dispersed into montane lakes along the Alps, Himalayas, Tibet, East Asia, and the North American Rocky Mountain ranges, with accelerated diversification rates outside Tian Shan. Climatically suitable regions for geographic lineages of G. lacustris were larger during cooling periods (LGM), but smaller during warming periods (Mid-Holocene). In the future (2070) scenario, potential distributions in the Himalayas, North Tibet, South Tibet and North America are predicted to expand, whereas ranges in East Asia, Europe and Tian Shan will decline. Our results suggest that Mid-Miocene-to-Pleistocene continuous cooling promoted multiple independent dispersal events out of Tian Shan due to increased availability of montane lakes via "budding" of lineages. Montane lakes are conduits through which cold-adapted amphipods globally dispersed, dominating circumboreal lakes. However, future climate warming is likely to force organisms to shift upward in altitude and northward in latitude, leading to a future change in local populations. These findings highlight the importance of conservation of montane lakes, especially in the context of climate change., (© 2022 John Wiley & Sons Ltd.)

We stated an error in our previous DNA analysis of Gammaridae. The right position of Zenkevitchia in the phylogenetic tree is close to marine and American Gammarus spp., far from the Dinaric troglobiotic Typhlogammarus group of genera. The morphologically diverse Zenkevitchia group supports the necessity of some paraphyly in systematics.

New genus and species of the family Niphargidae (Crustacea: Amphipoda: Senticaudata), Chaetoniphargus lubuskensis gen. nov., sp. nov. from the subterranean waters of cave Lubuška jama near Hajdučki Kukovi in the western part of Velebit Mt. in Croatia, is described and figured, and its relations to other genera and species of the family Niphargidae is discussed. The new taxon is a small niphargid, similar to Niphargobates. The list of all known genera of family Niphargidae is given and key to the genera of this family is presented.

The family Moitessieriidae includes minute dioecious gastropods exclusively inhabiting subterranean waters, including thermal ones. Only empty shells were collected in most species, the vast majority of them are described from their gross shell morphology alone. Several visits to a site are usually required to obtain at least some living individuals. High variability in shell morphology and the lack of diagnostic features, coupled with anticipated high levels of endemism, has resulted in a long list of nominal moitessierid species. Type specimens stored as empty shells omit unambiguous identification and delimitation of species boundaries. Due to inaccessibility of cave animals and consequent lack of material suitable for molecular analysis, the phylogenetic relationships, as well as the taxonomy of the family at genus/species level, are far from being understood. The anatomy of the family is also poorly known and provided only for a few taxa. The distinctness of the Moitessieriidae has sometimes been questioned, and their monophyly not proved. Twelve species of the Balkan Moitessieriidae are considered: two species of Paladilhiopsis, two species of Bythiospeum, six species of Iglica, Costellina turrita and Lanzaia bosnica. The shell morphology of each species, as well as the reproductive system of Paladilhiopsis and Iglica, were analysed. DNA sequences of nuclear histone H3, ribosomal 18S, ribosomal 28S and mitochondrial cytochrome oxidase subunit I (COI) were applied to infer phylogenetic relationships among the taxa. The sequences of Bythiospeum from GenBank have been used to infer relationships between Bythiospeum and Paladilhiopsis that were recently synonymized. Paladilhiopsis and Iglica are distinct, but closely related genera, as is the genus Bythiospeum, which does not occur in the Balkans. Its relationships with both former taxa remain unresolved. The Moitessieriidae are clearly distinct from all other families of the Truncatelloidea, however, their monophyly remains doubtful.

Three new species of the family Gammaridae-Gammarus troglomorphus, sp. n., G. parvioculatus, sp. n. from Lebap Province of Turkmenistan and Tadzocrangonyx alaicus, sp. n. from Batken Region of Kyrgyzstan are described and illustrated. Morphological studies of a closely related Turkmenistan population of G. cf. subaequalis-Garlyk, probably conspecific with Gammarus subaequalis Martynov, 1935 was provided. The affinity of new species to concerned taxa is discussed. To define phylogenetic position of mentioned species DNA barcode data are obtained. Gammarus troglomorphus and G. parvioculatus are close neighbors but exceedingly different morphologically. Gammarus troglomorphus is a troglobiont; G. parvioculatus is an eutroglophile, but with exception of slightly smaller eyes, not troglomorph. Both found only within small areas in the extreme East of Turkmenistan. Gammarus cf. subaequalis-Garlyk seems to extend from the same region far into the eastern Kyrgyzstan.

Turcolana Argano & Pesce, 1980 is the isopod genus occurring in freshwater and brackish groundwater environments around the eastern Mediterranean. In this study, a revised diagnosis of the genus, an updated map of species distribution and a key to species are presented. The first cave dwelling species is described from the Melissotrypa Cave in central Greece, a highly troglomorphic Turcolana lepturoides sp. nov. Its principal distinguishing characters are elongated appendages: styliform uropods (twice as long as pleotelson), antennae (half as long as body) and pereopods (e.g. pereopod 7 45% as long as body). Mitochondrial DNA sequences (16S rRNA, 12S rRNA) of the new species are provided.

Background: Several papers described the structure of curcumin and some other derivatives in solid and in solution. In the crystal structure of curcumin, the enol H atom is located symmetrically between both oxygen atoms of the enolone fragment with an O···O distance of 2.455 Å, which is characteristic for symmetrical H-bonds. In the solution, the geometry of the enolone fragment is attributed to the inherent disorder of the local environment, which solvates one of the basic sites better than the other, stabilizing one tautomer over the other. In this paper, how the position of methoxy groups in dimethoxy curcuminoids influence the conformation of molecules and how the halogen atoms change it when they are bonded at α-position in keto-enol part of molecules is described., Results: Six isomers of dimethoxy curcuminoids were prepared. Conformations in solid state, which were determined by X-ray single crystallography and 1H MAS and 13C CPMAS NMR measurements, depend on the position of methoxy groups in curcuminoid molecules. In solution, a fast equilibrium between both keto-enol forms exists. A theoretical calculation finding shows that the position of methoxy groups changes the energy of HOMO and LUMO. An efficient protocol for the highly regioselective bromination and chlorination leading to α-halogenated product has been developed. All α-halogenated compounds are present mainly in cis keto-enol form., Conclusions: The structures in solid state of dimethoxy curcuminoids depend on the position of methoxy groups. The NMR data of crystalline solid samples of 3,4-diOCH3 derivative, XRD measurements and X-ray structures lead us to the conclusion that polymorphism exists in solids. The same conclusion can be done for 3,5-diOCH3 derivative. In solution, dimethoxy curcuminoids are present in the forms that can be described as the coexistence of two equivalent tautomers being in fast equilibrium. The position of methoxy groups has a small influence on the enolic hydrogen bond. Theoretical calculations show that the energy gap between HOMO and LUMO depend on the position of methoxy groups and are lower in solution. Chlorination and bromination on α-position of 1,3-diketone moiety do not change the preferential form being cis keto-enol as in parent compounds.

Current theory predicts that a shift to a new habitat would increase the rate of diversification, while as lineages evolve into multiple species, intensified competition would decrease the rate of diversification. We used Holarctic amphipods of the genus Gammarus to test this hypothesis. We sequenced four genes (5,088 bp) for 289 samples representing 115 Gammarus species. A phylogenetic analysis showed that Gammarus originated from the Tethyan region with a saline ancestry in the Paleocene, and later colonized the freshwater habitat in the Middle Eocene. Ancestral range reconstruction and diversification mode analysis combined with paleogeological and paleoclimatic evidence suggested that the habitat shift from saline to freshwater led to an increased diversification rate. The saline lineage of Gammarus dispersed to both sides of the Atlantic at 55 million years ago (Ma), because of the few barriers between the Tethys and the Atlantic, and diversified throughout its evolutionary history with a constant diversification rate [0.04 species per million years (sp/My)]. The freshwater Gammarus, however, underwent a rapid diversification phase (0.11 sp/My) until the Middle Miocene, and lineages successively diversified across Eurasia via vicariance process likely driven by changes of the Tethys and landmass. In particular, the freshwater Gammarus lacustris and Gammarus balcanicus lineages had a relatively high diversification shift, corresponding to the regression of the Paratethys Sea and the continentalization of Eurasian lands during the Miocene period. Subsequently (14 Ma), the diversification rate of the freshwater Gammarus decreased to 0.05 and again to 0.01 sp/My. The genus Gammarus provides an excellent aquatic case supporting the hypothesis that ecological opportunities promote diversification.

The photochemical behavior of 2-halo-substituted 1,3-diarylpropan-1,3-dione strongly depends on the nature of the halogen atom bonded and the presence of electron-donor groups on the phenyl ring. In the case of 2-chloro-1,3-diphenylpropan-1,3-dione and 1-(3,5-dimethoxyphenyl)-3-phenylpropan-1,3-dione, cyclization to flavones was the sole reaction pathway, whereas in the case of 2-chloro-1,3-di(4-methoxyphenyl)propan-1,3-dione, only products derived from alpha-cleavage were observed. 2-Fluoro derivatives of 1,3-diarylpropan-1,3-diones were photostable; on the other hand, 2-chloro-2-fluoro derivates resulted in 3-fluoroflavones.

3',5'-dimethoxybenzoin (DMB) is an important photoremovable protecting group. The primary photoreactions of DMB acetate and fluoride following photoexcitation by a subpicosecond laser flash were investigated by pump-probe spectroscopy. The primary photoproduct is identified as a preoxetane biradical intermediate that decays by different pathways depending on solvent polarity. In polar solvents (acetonitrile, water), the biradical decays by releasing acetate or fluoride with a lifetime of about 2 ns. Thus, DMB is an excellent protecting group for the investigation of fast processes such as protein folding.