Synthesis of Functionalized 3H-pyrrolo-[1,2,3-de] Quinoxalines via Gold-Catalyzed Intramolecular Hydroamination of Alkynes.

A gold-catalyzed protocol to obtain functionalized 3H-pyrrolo [1,2,3-de] quinoxalines from suitable substituted N-alkynyl indoles has been proposed. The mild reaction conditions were revealed to be compatible with different functional groups, including halogen, alkoxyl, cyano, ketone, and ester, allowing the isolation of title compounds with yields from good to high. A reaction mechanism has been proposed, and theoretical calculations have been provided to rationalize the final step of the hypothesized reaction mechanism. As quinoxaline-containing polycyclic compounds, this class of molecules may represent a valuable template in medicinal chemistry and material science.

One-pot synthesis of dihydroquinolones by sequential reactions of o-aminobenzyl alcohol derivatives with Meldrum's acids.

The functionalized 3,4-dihydroquinolin-2-one nucleus has been assembled in good to high yields through the sequential reaction of readily available N-Ts-o-aminobenzyl alcohols with 5-substituted Meldrum's acid derivatives under mild basic conditions. Highly diastereoselective synthesis of 3-substituted-4-phenyl-1-tosyl-3,4-dihydroquinolin-2(1H)-ones was accomplished from N-(2-(hydroxy(phenyl)methyl)phenyl)-4-methylbenzenesulfonamide under the same reaction conditions. Regarding the reaction mechanism, we hypothesized that the formation of dihydroquinolones proceeds through the in situ generation of aza-o-QMs followed by conjugate addition of enolate/cyclization/elimination of acetone and CO2.

Synthesis of 4-Substituted-1,2-Dihydroquinolines by Means of Gold-Catalyzed Intramolecular Hydroarylation Reaction of N-Ethoxycarbonyl-N-Propargylanilines.

An alternative Au(I)-catalyzed synthetic route to functionalized 1,2-dihydroquinolines is reported. This novel approach is based on the use of N-ethoxycarbonyl protected-N-propargylanilines as building blocks that rapidly undergo the IMHA reaction affording the 6-endo cyclization product in good to high yields. In the presence of N-ethoxycarbonyl-N-propargyl-meta-substituted anilines, the regiodivergent cyclization at the ortho-/para-position is achieved by the means of catalyst fine tuning.

DNA Taxonomy Confirms the Identity of the Widely-Disjunct Mediterranean and Atlantic Populations of the Tufted Ghost Crab Ocypode cursor (Crustacea: Decapoda: Ocypodidae).

The distribution area of the tufted ghost crab Ocypode cursor includes two widely separate sub-areas, i.e. the tropical and subtropical Atlantic coasts of Africa and Macaronesia, and the central-eastern Mediterranean basin. The current disjunct distribution of the species is possibly the remnant of a previous wider and continuous distribution area that was fragmented during the Pleistocene, with the disappearance of the species from the temperate Atlantic Ocean and the western Mediterranean basin, and its survival in the warmer areas of the eastern Atlantic and the Mediterranean Sea. Such disjunction is thus compatible with an ancient isolation between the Mediterranean and Atlantic populations of the species, which could in fact constitute two well-characterized independent evolutionary lineages, or even two cryptic species. Unexpectedly, the sequencing of a fragment of the mtDNA COI gene from Mediterranean and Atlantic Ocypode cursor allopatric populations showed the cohesion of the species throughout its distribution range, and the nesting of Mediterranean populations within the single Atlantic population studied. This pattern is hereby tentatively ascribed to an incomplete lineage sorting due to the large population sizes of both the Atlantic and Mediterranean subpopulations of the species. The current westward expansion of the species in the Mediterranean Sea originating from the Levantine basin, due to ongoing regional sea warming, follows a typical phalanx dispersal mode.

The DNA barcoding of Mediterranean combtooth blennies suggests the paraphyly of some taxa (Perciformes, Blenniidae).

A dataset including novel and publicly available mtDNA COI sequences of 14 Mediterranean combtooth blenny species belonging to nine genera was assembled in order to provide a reference dataset for DNA barcoding studies. Some inconsistencies in the current taxonomy of some genera were observed. In particular, the monophyly of the genera Parablennius and Salaria is not supported by the present dataset and the absence of reciprocal monophyly between the morphospecies Salaria basilisca and S. pavo questions their status and stresses the need of a revision of the genus Salaria.

On the Occurrence of the Invasive Freshwater Limpet Ferrissia californica (Mollusca: Gastropoda: Planorbidae) in Japan.

Two members of the genus Ferrissia Walker, 1903, namely Ferrissia nipponica (Kuroda, 1949) and F. japonica Habe and Burch, 1965, have been reported to occur in Japan, but due to a lack of molecular data, doubts have been expressed as to their validity. Furthermore, the possible presence of allochthonous Ferrissia taxa has been stated under a variety of names, so that even now there is no consensus on their presence and identity. Recently, freshwater limpets belonging to the planorbid genus Ferrissia were collected in an irrigation trough on the Izu island of Hachijojima, Tokyo, Japan. Molecular identification of the collected specimens, based on a fragment of the large ribosomal subunit 16S (mtDNA), unequivocally proved that they belong to the allochthonous, invasive species Ferrissia californica (Rowell, 1863), the protagonist of a relentless cryptic invasion worldwide to which can Japan can now be added. The shells of the collected specimens show some scars, which bear testimony to the fragile nature of the shell and, possibly, to the paucity of calcium in the inland waters of Hachijojima.

The good, the bad and the ugly: Emys trinacris, Placobdella costata and Haemogregarina stepanowi in Sicily (Testudines, Annelida and Apicomplexa).

Endemic Sicilian pond turtles Emys trinacris Fritz, Fattizzo, Guicking, Tripepi, Pennisi, Lenk, Joger et Wink were examined for the presence of haemogregarine parasites. The presence of haemogregarines, occurring mainly in the microgametocyte stage (13.2 ± 0.12 µm in length and 6.4 ± 0.52 µm in width), was observed in approximately 9% of the sampled E. trinacris. Based on the observed morphology and on the sequencing of nuclear 18S rDNA, we identified the parasite as Haemogregarina stepanowi Danilewsky, 1885. Morphometric study of uninfected and infected red blood cells has shown that H. stepanowi induces different changes in erythrocyte shape depending on the infective stage. The differential count of leukocytes in specimens infected with H. stepanowi showed no significant difference compared with healthy specimens. However, considering the health problems which might be induced by H. stepanowi in the closely related European pond turtle Emys orbicularis (Linneaus), monitoring of the health status of the infected Sicilian populations of E. trinacris is desirable. The restricted distribution of populations of Emys infected with haemogregarines in Sicily is quite puzzling and the possible human-mediated introduction of the parasite in Sicily is briefly discussed.

Distribution and Genetic Lineages of the Craspedacusta sowerbii Species Complex (Cnidaria, Olindiidae) in Italy.

Olindiid freshwater jellyfishes of the genus Craspedacusta Lankester, 1880 are native to eastern Asia; however, some species within the genus have been introduced worldwide and are nowadays present in all continents except Antarctica. To date, there is no consensus regarding the taxonomy within the genus Craspedacusta due to the morphological plasticity of the medusa stages. The species Craspedacusta sowerbii Lankester, 1880 was first recorded in Italy in 1946, and until 2017, sightings of the jellyfish Craspedacusta were reported for 40 water bodies. Here, we shed new light on the presence of the freshwater jellyfishes belonging to the genus Craspedacusta across the Italian peninsula, Sardinia, and Sicily. First, we report 21 new observations of this non-native taxon, of which eighteen refer to medusae sightings, two to environmental DNA sequencing, and one to the finding of polyps. Then, we investigate the molecular diversity of collected Craspedacusta specimens, using a Bayesian analysis of sequences of the mitochondrial gene encoding for Cytochrome c Oxidase Subunit I (mtDNA COI). Our molecular analysis shows the presence of two distinctive genetic lineages: (i) a group that comprises sequences obtained from populations ranging from central to northern Italy; (ii) a group that comprises three populations from northern Italy-i.e., those from the Lake Levico, the Lake Santo of Monte Terlago, and the Lake Endine-and the single known Sicilian population. We also report for the first time a mtDNA COI sequence obtained from a Craspedacusta medusa collected in Spain.

Overlooked cryptic endemism in copepods: systematics and natural history of the calanoid subgenus Occidodiaptomus Borutzky 1991 (Copepoda, Calanoida, Diaptomidae).

Our comprehension of the phylogeny and diversity of most inland-water crustaceans is currently hampered by their pronounced morphological bradytely, which contributed to the affirmation of the "Cosmopolitanism Paradigm" of freshwater taxa. However, growing evidence of the existence of cryptic diversity and molecular regionalism is available for calanoid copepods, thus stressing the need for careful morphological and molecular studies in order to soundly investigate the systematics, diversity and distribution patterns of the group. Diaptomid copepods were here chosen as model taxa, and the morphological and molecular diversity of the species belonging to the west-Mediterranean diaptomid subgenus Occidodiaptomus were investigated with the aim of comparing the patterns of morphological and molecular evolution in freshwater copepods. Three species currently lumped under the binomen Hemidiaptomus (Occidodiaptomus) ingens and two highly divergent clades within H. (O.) roubaui were distinguished, thus showing an apparent discordance between the molecular distances recorded and Occidodiaptomus morphological homogeneity, and highlighting a noteworthy decoupling between the morphological and molecular diversity in the subgenus. Current Occidodiaptomus diversity pattern is ascribed to a combined effect of ancient vicariance and recent dispersal events. It is stressed that the lack of sound calibration points for the molecular clock makes it difficult to soundly temporally frame the diversification events of interest in the taxon studied, and thus to asses the role of morphological bradytely and of accelerated molecular evolutionary rates in shaping the current diversity of the group.

Molecular phylogeny of the Notostraca.

We used a combined analysis of one nuclear (28S rDNA) and three mitochondrial markers (COI, 12S rDNA, 16S rDNA) to infer the molecular phylogeny of the Notostraca, represented by samples from the six continents that are inhabited by this group of branchiopod crustaceans. Our results confirm the monophyly of both extant notostracan genera Triops and Lepidurus with good support in model based and maximum parsimony analyses. We used branchiopod fossils as a calibration to infer divergence times among notostracan lineages and accounted for rate heterogeneity among lineages by applying relaxed-clock models. Our divergence date estimates indicate an initial diversification into the genera Triops and Lepidurus in the Mesozoic, most likely at a minimum age of 152.3-233.5 Ma, i.e., in the Triassic or Jurassic. Implications for the interpretation of fossils and the evolution of notostracan morphology are discussed. We further use the divergence date estimates to formulate a biogeographic hypothesis that explains distributions of extant lineages predominantly by overland dispersal routes. We identified an additional hitherto unrecognised highly diverged lineage within Lepidurus apus lubbocki and three additional previously unknown major lineages within Triops. Within T. granarius we found deep differentiation, with representatives distributed among three major phylogenetic lineages. One of these major lineages comprises T. cancriformis, the T. mauritanicus species group and two hitherto unrecognised T. granarius lineages. Samples that were morphologically identified as T. granarius diverged from the most basal nodes within this major lineage, and divergence dates suggested an approximate age of 23.7-49.6 Ma for T. cancriformis, indicating the need for a taxonomic revision of Triassic and Permian fossils that are currently attributed to the extant T. cancriformis. We thus elevate T. cancriformis minor to full species status as Triops minorTrusheim, 1938 and include in this species the additional Upper Triassic samples that were attributed to T. cancriformis. We further elevate T. cancriformis permiensis to full species status as Triops permiensisGand et al., 1997.

Disentangling cryptic species in Parastenocarididae (Copepoda: Harpacticoida) with an integrative approach: the case of Stammericaris similior sp. nov. and Stammericaris destillans Bruno & Cottarelli 2017.

Stammericaris similior sp. nov. is described combining light microscopy, scanning electron microscopy, and genetic barcoding. The new species was collected from rimstone pools in Scrivilleri Cave, a cave in Sicily with so far unexplored microcrustacean fauna. The new species is particularly interesting because it is morphologically very similar to Stammericaris destillans, an epikarstic parastenocaridid endemic to a different Sicilian cave; however, the phylogenetic analysis based on the mitochondrial COI gene of sixteen parastenocaridids shows that these two Stammericaris are two distinct species, with an uncorrected p-distance of 22.9, and the sequences of Stammericaris similior sp. nov. cluster together in a well-supported monophyletic clade, with two different haplotypes. To our knowledge, the presence of different species of almost identical morphology had not been recorded before for the genus Stammericaris. The integrated molecular and morphological analysis, the latter conducted with the support of SEM, allows disentangling the affinities of the new species and identifying a few distinctive characters: the males of the new species are characterized by the caudal rami shorter than the anal somite; the morphology of the P3, which is thin and slightly arched, with three proximal spinules on exp-1; the peculiar structure of the P4 enp; the P4 basis ornamented with two spinules of different length, the one closest to the endopod being the shortest one, and a half-moon shaped lamella. The new species differs from S. destillans for its larger size, the presence of: three spinules, instead of two, on the P3 exp-1; the half-moon shaped lamella on the P4 basis; a row of spinules along the inner margin of P4 exp-1. We also provide data on the ecology and distribution of the new species, a list of the other copepod species collected, and a dichotomic key for the males of all species presently assigned to the genus.

Morphological and molecular diversity patterns of the genus Tropodiaptomus Kiefer, 1932 (Copepoda, Calanoida, Diaptomidae) in Thailand.

Tropodiaptomus is one of the most specious genera in the family Diaptomidae, but it is often rare in terms of distribution and abundance. Moreover, Tropodiaptomus species show a noteworthy variability in some of the morphological characters considered of prime importance in diaptomid taxonomy, and the presence of cryptic or pseudocryptic species is likely. Thus, through a geographically-wide sampling in Thailand, we aimed to investigate the local diversity of the genus and to compare the morphological and molecular diversity pattern based on mitochondrial and nuclear genes sequences. DNA taxonomy was also implemented in order to check whether the Tropodiaptomus lineages were independent species according to the "evolutionary genetic species concept". Six Tropodiaptomus morphospecies were found, three of which are putative species new to Science pending a formal description. The finding of such a high incidence of undescribed species stresses the existence of a significant "Linnean shortfall" affecting Thai diaptomids. The molecular results showed that most of the studied species could be identified consistently with their morphology-based taxonomy. However, Tropodiaptomus vicinus and T. cf. lanaonus showed a high level of genetic diversity, suggesting that traditional morphological techniques might be inadequate for correctly assessing their taxonomical status.

A redescription of the Mediterranean endemic cladoceran Daphnia chevreuxi Richard, 1896 (Cladocera: Daphniidae).

The Mediterranean area is a biodiversity and endemism hotspot. Circum-Mediterranean taxa are known among different hydrobionts, including the water fleas. Some Mediterranean endemic cladoceran taxa have been described or redescribed according to modern taxonomical standards, but accurate drawings are missing for others. Here we redescribe the Mediterranean endemic Daphnia chevreuxi Richard, 1896 (Crustacea: Cladocera) and briefly review available data on its distribution and ecology. The species is confirmed to be a typical inhabitant of the temporary ponds of the central Mediterranean area, whereas its populations from the eastern Balkans and the Middle East should be studied in order to check for their actual identity. We conclude that the Mediterranean area is an example of a well-studied region as Cladocera are concerned, but the study of other regions is necessary in order to understand better the cladoceran diversity and distribution patterns in Eurasia.

The European Pine Marten Martes martes (Linnaeus, 1758) Is Autochthonous in Sicily and Constitutes a Well-Characterised Major Phylogroup within the Species (Carnivora, Mustelidae).

No molecular data are currently available for the Sicilian populations of the European pine marten Martes martes, thus preventing any sound inference about its native or non-native status on the island, as well as the local phylogeography of the species. In order to investigate these issues, we sequenced two mtDNA markers in road-killed specimens collected in Sicily. Both markers consistently demonstrated the existence of a well-characterised Sicilian clade of the species, which is endemic to the island and constitutes the sister group of a clade including the Mediterranean and Central-North European major phylogroups of the European pine marten. Such evidence supports the autochthony of Martes martes in Sicily and points to a natural Pleistocene colonisation of the island followed by isolation. The occurrence of a, to date undetected, major phylogroup of the species in Sicily calls for the dedicated monitoring of the Sicilian populations of the species in order to preserve this evolutionarily significant unit.

Pleistocene allopatric differentiation followed by recent range expansion explains the distribution and molecular diversity of two congeneric crustacean species in the Palaearctic.

Pleistocene glaciations had a tremendous impact on the biota across the Palaearctic, resulting in strong phylogeographic signals of range contraction and rapid postglacial recolonization of the deglaciated areas. Here, we explore the diversity patterns and history of two sibling species of passively dispersing taxa typical of temporary ponds, fairy shrimps (Anostraca). We combine mitochondrial (COI) and nuclear (ITS2 and 18S) markers to conduct a range-wide phylogeographic study including 56 populations of Branchinecta ferox and Branchinecta orientalis in the Palaearctic. Specifically, we investigate whether their largely overlapping ranges in Europe resulted from allopatric differentiation in separate glacial refugia followed by a secondary contact and reconstruct their postglacial recolonization from the inhabited refugia. Our results suggest the existence of distinct refugia for the two species, with genetic divergence among intraspecific lineages consistent with late Pleistocene glacial cycles. While B. ferox lineages originated from Mediterranean refugia, the origin of B. orientalis lineages was possibly located on the Pannonian Plain. We showed that most dispersal events predominantly happened within 100 km, coupled with several recent long-distance events (> 1000 km). Hence the regional habitat density of suitable habitats in Central Europe is possibly a key to the co-existence of the two species. Overall, our study illustrates how isolation in combination with stochastic effects linked to glacial periods are important drivers of the allopatric differentiation of Palaearctic taxa.

Palladium-catalyzed Tsuji-Trost-type reaction of benzofuran-2-ylmethyl acetates with nucleophiles.

The palladium-catalyzed benzylic-like nucleophilic substitution of benzofuran-2-ylmethyl acetate with N, S, O and C soft nucleophiles has been investigated. The success of the reaction is dramatically influenced by the choice of catalytic system: with nitrogen based nucleophiles the reaction works well with Pd2(dba)3/dppf, while with sulfur, oxygen and carbo-nucleophiles [Pd(η3-C3H5)Cl]2/XPhos is more efficient. The regiochemical outcome shows that the nucleophilic substitution occurs only on the benzylic position of the η3-(benzofuryl)methyl complex. The high to excellent yields and the simplicity of the experimental procedure make this protocol a versatile synthetic tool for the preparation of 2-substituted benzo[b]furans.

Diversity and taxonomy of the genus Unio Philipsson in Italy, with the designation of a neotype for Unio elongatulus C. Pfeiffer (Mollusca, Bivalvia, Unionidae).

Novel Unio spp. populations from Slovenia, the Italian peninsula, Sardinia and Sicily were genetically analysed in order to define the distribution and diversity of the genus Unio in Italy and neighbouring areas. The presence of two primarily allopatric autochtonous species, Unio elongatulus Pfeiffer, and Unio mancus Lamarck, is confirmed for the Italian peninsula, Corsica, Sardinia, and Sicily.Autochthonous populations of Unio elongatulus are present in the peri-Adriatic drainages of the Italian and Balkan peninsulas, south as far as the Ofanto River (Apulia, Italy) and Lake Skadar (Albania), while its presence in the Tyrrhenian rivers of Tuscany is likely due to anthropogenic introduction events. Conversely, Unio mancus turtonii Payraudeau, an endangered peri-Tyrrhenian taxon, was found with autochthonous populations in the Apennine-Tyrrhenian drainages of peninsular Italy, eastern Mediterranean France, Corsica, Sardinia and Sicily, while the actual autochthony of the single population found in the Ionian basin of the Italian peninsula (Bradano River, Basilicata) deserves further investigation. The Italian population of U. mancus requienii Michaud, reported from Lake Montepulciano is to be considered allochthonous.The binomen U. elongatulus, although widely used in the recent scientific literature, was, to date, assigned to a doubtful species, because its type locality includes a large area inhabited by different Unio taxa, which are not clearly distinguishable by their shell alone; furthermore, no type material is present in historical collections. To retain the recently-used name, a new restricted type locality is established, where only U. elongatulus lives, and a neotype is designated. The validity of the subspecies of Unio mancus is also discussed and confirmed.Finally, hypotheses on the origin of Italian mussels, and considerations on their conservation status are discussed.

An annotated checklist of freshwater Copepoda (Crustacea, Hexanauplia) from continental Ecuador and the Galapagos Archipelago.

An annotated checklist of the free-living freshwater Copepoda recorded in different regions in Ecuador (including the Amazon, the Andes, the coastal region, and the Galapagos Islands) is here provided. We revised all published records, critically evaluated the validity of each taxon and provided short taxonomic and biogeographical remarks for each one. A total of 27 taxa have been reported, including species and records at the generic level only. The species and taxa identified only up to the generic level belong to five families and 14 genera. The Cyclopoida is the most diverse group with 16 records belonging to species (or identified to the generic level only) and eight genera, followed by the Harpacticoida with six species, one identification to the generic level only, and four genera, and Calanoida with four species belonging to two genera. A total of 18 taxa are recorded for the Andes. Six have been recorded in the Amazon, two are recorded for the coastal region, and six for the Galapagos. One species is shared between the Amazon and the Andes. One species is shared between the coastal region and the Amazon. Seventeen are only reported from the Andes and four are only reported from the Amazon. At the current status of the knowledge, any attempt to analyze and generalize distributional patterns of copepods in Ecuador is premature due to the scarcity of available information, and evidently there is an urgent need for more extensive field collections. A few working hypothesis for future studies are identified.

Revision of Recent and fossil Mixtacandona Klie 1938 (Ostracoda, Candonidae) from Italy, with description of a new species.

Although studies on ostracods dwelling in inland subterranean habitats of Italy have increased in the last decades, highlighting a considerable taxonomic diversity, available information is still far from sufficient to understand phylogenetic and biogeographic relationships. Mixtacandona Klie 1938 is one of the most common and diverse genera of ostracods in subterranean waters. Of the 20 living recognized species in the genus, all stygobiontic and restricted to the Palearctic Region, four are known for the Italian peninsula and Sardinia, one of which exclusively as fossil. Several other Recent and fossil taxa attributable to Mixtacandona, but identified at supraspecific level, have been reported from various Italian regions. Here we report Mixtacandona idrisi n. sp., collected in a cave near Palermo, the first cavernicolous ostracod from the island of Sicily; in addition, the description of material recently collected in northern Italy, consisting of M. laisi and of a putative new species that has been tentatively allocated to the same genus, is given. Mixtacandona (Trapezicandona) italica Karanovic & Pesce, 2000 is tentatively treated as a junior synonym of M. talianae Gliozzi & Mazzini, 1998. The validity of species-groups within Mixtacandona is discussed. Relying on the results of this study, the authors argue for the need of a comprehensive revision of the genus.