An overview of the Hygrobates fluviatilis complex (Acariformes, Hydrachnidia, Hygrobatidae), with the description of a new species from Portugal and the first record of H. persicus from Cyprus

Water mites of the Hygrobates fluviatilis-complex are often the most ubiquitous and usually the most abundant water mite group in the running waters of the Palaearctic and Balkans. In the present study we used an integrative taxonomic approach by applying partial COI sequences (DNA-barcodes) to describe two new species of the fluviatilis-complex from the Balkan Peninsula, H. balcanicus Pešić sp. nov. (Bulgaria, North Macedonia, Greece) and H. mediterraneus Pešić sp. nov. (Montenegro). Phylogenetic analysis based on COI data placed H. balcanicus sp. nov. as the sister species of the clade grouping H. turcicus Pešić, Esen & Dabert, 2017 and H. ulii Pešić et al. 2019 whereas H. mediterraneus sp. nov. is placed as the sister species of H. grabowskii Pešić et al. 2019, with about 20% and 13% K2P divergence, respectively, between the closest relatives.

We describe a new species of Eupsophus from the Coastal Range of Southern Chile. The new taxon can be differentiated using an integrative taxonomic approach that includes advertisement call, reproductive mode, tadpoles, and mtDNA D-loop sequences. Based on karyological characters, the new species is assigned to the roseus Group (2N=30), and differs from all described species therein by having early winter breeding season, terrestrial tadpoles and its advertisement call with spectral elements reaching the 20 kHz. Phylogenetic analyses based on mitochondrial data place the new species as sister taxon of E. migueli. The discovery of this new species highlights the importance of the multisource approaches to discover cryptic diversity as well as to establish the basis for prioritizing policies and conservation efforts on Chilean batrachofauna.

The Mediterranean islands are known as natural laboratories of evolution with a high level of endemic biodiversity.However, most biodiversity assessments have focused mainly on terrestrial and marine fauna, leaving the freshwater animals aside.Crete is one of the largest islands in the Mediterranean Basin, with a long history of isolation from the continental mainland.Gammarid amphipods are often dominant in macrozoobenthic communities in European inland waters.They are widely used in biomonitoring and exotoxicological studies.Herein, we describe Gammarus plaitisi sp.nov., endemic to Cretan streams, based on morphological characters and a set of molecular species delimitation methods using mitochondrial cytochrome oxidase subunit I and 16S rRNA genes as well as nuclear 28S rDNA, ITS1 and EF1-alpha genes.The divergence of the new species is strongly connected with the geological history of the island supporting its continental origin.

The Mediterranean islands are known as natural laboratories of evolution with a high level of endemic biodiversity. However, most biodiversity assessments have focused mainly on terrestrial and marine fauna, leaving the freshwater animals aside. Crete is one of the largest islands in the Mediterranean Basin, with a long history of isolation from the continental mainland. Gammarid amphipods are often dominant in macrozoobenthic communities in European inland waters. They are widely used in biomonitoring and exotoxicological studies. Herein, we describe Gammarus plaitisi sp. nov., endemic to Cretan streams, based on morphological characters and a set of molecular species delimitation methods using mitochondrial cytochrome oxidase subunit I and 16S rRNA genes as well as nuclear 28S rDNA, ITS1 and EF1-alpha genes. The divergence of the new species is strongly connected with the geological history of the island supporting its continental origin.

An integrative taxonomic framework for the study of the genus <i>Ciona</i> (Ascidiacea) and description of a new species, <i>Ciona intermedia</i>

Species of the allocreadiid genus Creptotrema are parasites of freshwater fishes in the Americas. Species in the genus possess one pair of muscular oral lobes on the oral sucker. Currently, the genus contains eight species, six distributed in South America, one in Middle America and one in North America. Genetic data are only available for the North American species, Creptotrema funduli, a parasite of fundulids originally described from Oneida Lake, New York State. In this study, we obtained 28S ribosomal DNA sequences of trematodes morphologically similar to Creptotrema agonostomi from the mountain mullet, Dajaus monticola, across a wide geographical range in Middle America. Our molecular phylogenetic analyses showed that (1) the genus Creptotrema, as currently conceived, is not monophyletic; (2) the allocreadiids in mountain mullets should be re-allocated in the genus Pseudoparacreptotrema; and (3) the allocreadiid trematodes from D. monticola across Middle America represent four morphologically similar species, three of which can be distinguished genetically. These three new species are described herein using an integrative taxonomy approach. We contend that accurate estimates of species diversity and phylogenetic relationships among allocreadiids, and most likely other species of trematodes, necessarily require an integrative taxonomy approach that should consider at least DNA sequences and scanning electron microscopy.

Simple SummaryDescribing global biodiversity involves identifying species and describing their distributions. The subterranean termite genus Reticulitermes represents an important group of wood-destroying organisms; however, little is known about their species-specific distribution across the three northern hemisphere continents where they are endemic. We combined several taxonomic methods to identify the species of over 4000 specimens in the first statewide survey of subterranean termites from Georgia, USA. The area surveyed, 153,900 km2, represents eco-regions typical of most of the southeast and eastern seaboard of the United States. There were three species, R. flavipes, R. virginicus, and R. nelsonae, found throughout Georgia. R. malletei was predominantly collected in the northern Piedmont soil province, while R. hageni, the least encountered species, was not collected from South Georgia. Our findings support the need for a taxonomic revision of the genus Reticulitermes, agreement on an appropriate integrated taxonomic approach for species determination, and should stimulate future research on diverse topics such as biodiversity, monitoring for these structural pests, and identifying their role in forest ecosystems.Reticulitermes subterranean termites are widely distributed ecosystem engineers and structural pests, yet describing their species distribution worldwide or regionally has been hindered by taxonomic uncertainties. Morphological plasticity confounds the use of taxonomic keys, while recent species descriptions and molecular techniques lacking taxonomic support have caused a muddle in interpreting the literature on Reticulitermes species distributions. We employed an integrative taxonomic approach combining behavioral, morphological, and molecular techniques to identify 4371 Reticulitermes samples to species. Five Reticulitermes species were collected from wood-on-ground at 1570 sites covering 153,900 km2 in the state of Georgia, USA. Three species were collected throughout Georgia, with R. flavipes identified from every one of the 159 counties. R. nelsonae was the second most frequently collected species, found in 128 counties, with R. virginicus third with 122. Two species had distributions confined to the northern part of the state. R. malletei was collected from 73 counties, while the least collected species, R. hageni, was found in 16. Results show that the most recently described species (R. nelsonae, 2012) is widely distributed and the second-most frequently encountered termite, representing 23% of all samples. The invasive species R. flavipes represented half of all the samples collected, while R. hageni, the least at less than 1%. A search of GenBank identified a number of accessions mismatched to a species designation resulting in the literature under-reporting the biodiversity of the genus. We, therefore, outline a path to standardize methods for species identification using an integrated taxonomic approach with appropriate barcodes for consistent identification across research teams worldwide. The data also illuminate new opportunities to examine questions related to the ecology, evolution, dispersal, and resource partitioning behaviors of these sympatric species across distinct geographical regions.

New insights into the evolution and biogeography of freshwater planarians on islands in the Tyrrhenian Sea, Western Mediterranean Basin, with the integrative description of a new endemic species from Corsica (Platyhelminthes: Tricladida: <i>Dugesia</i>)

\n In this work, the integrative taxonomy approach was performed to understand species limits and patterns of diversification in two genera of orizomyine rodents (Cerradomys and Oligoryzomys). Therefore, molecular markers with distinct evolutionary rates were used with different approaches (phylogeny, coalescent-based species delimitation, DNA barcoding, phylogeography, molecular dating). Classic and molecular cytogenetic analyzes were performed, contributing to cytotaxonomy and revealing chromosomal evolution. This work is divided into four chapters, including a brief introduction (Chapter 1). In Chapter 2, the integrative taxonomy approach was used to study the genus Cerradomys, based on cytogenetic and molecular data. The results revealed that cytogenetics is important in the recognition of all described species (cytotaxonomy). Phylogenetic reconstruction showed that internal relationships are well supported, with the exception of C. subflavus and C. goytaca, which are not reciprocally monophyletic. Following the integrative taxonomy, in which species limits are based on the congruence of methods, this work recognizes and reiterates the eight Cerradomys species described so far. We suggest a taxonomic revision in C. langguthi and C. subflavus, since both may represent species-complex or in process of speciation. Times of divergence show that Cerradomys is a recent genus, with speciation events occurred mainly in the Pleistocene. In Chapter 3, classic and molecular cytogenetics (Fluorescence in situ hybridization - FISH with telomeric and Oligoryzomys moojeni probes) were used to study chromosomal evolution in Cerradomys, based on the molecular phylogeny obtained in Chapter 2. Chromosome painting revealed extensive chromosome reshuffling in Cerradomys. Species with the highest diploid numbers showed exclusively telomeric signals whereas interstitial telomeric signals (ITS) were observed in the species with the lowest diploid numbers. Comparisons of chromosome painting with molecular phylogeny data corroborate the hypothesis that ITS, in this case, are remnants of telomeres. Nevertheless, other chromosomal rearrangements were detected with absence of ITS, indicating that these sequences may have been lost in the process of chromosomal breakages, evidencing that there was both retention and loss of ITS along the karyotypic evolution of the genus. In addition, complex rearrangements were detected between the karyotypes of C. goytaca and C. subflavus, reiterating that these two species are distinct, since hybrids probably would not be viable due to meiotic problems. In Chapter 4, aiming to recover the evolutionary history and species limits of Oligoryzomys, molecular phylogeny studies were integrated into cytogenetic data. The genus was monophyletic, but the internal relations had low support. The compilation of phylogenetic, chromosomal data and geographic distribution (interdisciplinarity) was important to understand species boundaries. Four lineages could not be related to any name and may be new species (Oligoryzomys A-D). Oligoryzomys flavescens was recovered paraphyletic in respect to O. fornesi. Oligoryzomys stramineus, O. microtis and O. nigripes were recovered in two well-structured clades each. In the case of the last two species, the subclades are probably related to exclusive karyotypes. In O. microtis, one subclade is composed of samples from the western Amazon region and the other with samples distributed in southern Amazon region, transition with Cerrado (2n=64, FN=64). In O. nigripes, one of the clades is composed of specimens from northeastern Brazil (2n=62, FN=78) and the other from central-south-southeast Brazil, Argentina, Paraguay and Uruguay (2n=62, FN=80-82). Phylogeographic results corroborate phylogenetic and cytogenetic data, revealing two distinctive phylogroups, consistent with incipient species. Chromosome data corroborate previous work and could be associated to the following names: O. mattogrossae, O. moojeni, O. chacoensis, O. stramineus, O. nigripes and O. flavescens, although the last two species should be reassessed. In addition, an undescribed karyotype is being reported for Oligoryzomys aff. utiaritensis (2n=70, FN=72), as well as new records in Brazil for four species. We suggest a taxonomic revision in O. microtis, O. flavescens and O. nigripes, as these species probably represent incipient or species-complex. In addition, samples related to Oligoryzomys aff. delicatus, Oligoryzomys aff. chacoensis, Oligoryzomys aff. rupestris and Oligoryzomys aff. Utiaritensis should be evaluated morphologically to confirm their identities. The results of this work corroborate the importance of interdisciplinary studies, since the rates of evolution differ according to each character\n

Zootaxa came as a new and innovative publication medium for taxonomy, amidst a scenario of devaluation of this important biological science. After 20 years, it has ascertained itself as one of the main journals in animal taxonomy. However, the contribution of the journal to the taxonomy of Mammalia (mammals), one of the most studied groups of animals with a long-standing, dedicated spectrum of specialized journals (mammalogy), could have been expected as minor. All the current and former editors of the Mammalia section of Zootaxa analyzed the relative contribution of the journal to the description of new species of mammals since 2001. We also analyzed the contribution of Zootaxa by taxon, geographic origin of taxa, and geographic origin of first authors. The taxonomic methodology of authors in species description is described as well as the temporal trends in publications and publication subjects. We highlight the editors' picks and eventually, the challenges for the future. We found that Zootaxa has had a significant contribution to mammalogy, being the second journal (the first being Journal of Mammalogy) in terms of number of new species described (76; 10.6% of the new mammalian species described between 2001 and 2020). The majority of the new species were described following an integrative taxonomic approach with at least two sources of data (86%). The analysis of published taxa, their geographic origin, and the country of origin of first authors shows a wide coverage and exhaustive representation, except for the species from the Nearctic. We conclude that Zootaxa has likely responded to a repressed demand for an additional taxonomic journal in mammalogy, with as possible appeals the absence of publication fees and an established publication speed. With 246 articles published in the past 20 years, the Mammalia section of Zootaxa embraces a large spectrum of systematic subjects going beyond alpha taxonomy. The challenges for the future are to encourage publications of authors from the African continent, still poorly represented, and from the palaeontology community, as the journal has been open to palaeontology since its early days.

Species delimitation and species concepts have been a matter of debate among biodiversity researchers in the last decades, resulting in integrative taxonomy approaches and the use of modern species concepts, such as the phylogenetic, evolutionary or general lineage species concepts. The discussion of subspecies status and concepts has been addressed much less extensively, with some researchers completely refraining from recognizing subspecies. However, allopatric insular populations that are particularly differentiated have traditionally been assigned subspecies status. We studied the molecular phylogeny and morphology of endemic Comoran tree snakes of the genus Lycodryas. Taking an integrative taxonomic approach, we used the concept of independent lines of evidence to discriminate between evidence for specific and subspecific status. Molecular (mtDNA) and morphological data provided sufficient evidence to support four different taxa within Comoran Lycodryas. In a revision of this group, we propose two species, each with two subspecies. We present a discussion of the strong sexual dichromatism unique to Comoran Lycodryas within the genus and related genera that may be explained by sexual selection in combination with the absence of major predators. Then, we discuss the effects of insular evolution and the “island rule” on morphological traits in Comoran Lycodryas and in Liophidium mayottensis, another snake endemic to the Comoros. The absence of larger-bodied snakes may have promoted an increase in body size and the number of dorsal scale rows in these species. Finally, we discuss the subspecies concept, its applications and its significance for integrative taxonomy and for limiting taxonomic inflation. We emphasize that taxon descriptions should be based on an integrative approach using several lines of evidence, preferably in combination with statements on the underlying species concepts or operational criteria, to increase the objectivity and comparability of descriptions.

In the present study we used an integrative taxonomic approach that combines morphological and DNA barcoding data to describe Hygrobates neotrigonicus sp. nov., a new species of H. trigonicus-complex from Bingöl Province (East Türkiye). The average K2P-distance between DNA-barcode sequences from the new species and two other European clades of H. trigonicus was estimated at 16.6 ±1.7% and 19.0±2.0% K2P, respectively.

The present work aims to expand the knowledge of the digenean species Prosogonotrema bilabiatum (Sclerodistomidae), a parasite of Chaetodipterus faber (Acanthuriformes) from Brazil, with an integrative taxonomic approach, using light microscopy, scanning electron microscopy, histology, and molecular biology. Forty-one digenean specimens were stained with hydrochloric carmine for morphological studies. Eleven parasites were dehydrated through a graded ethanol series, critical point dried with carbon dioxide, and coated with gold for scanning electron microscopy analysis. Four specimens were processed following histological routine and stained with hematoxylin and eosin and Gomori trichrome. DNA extracted was amplified using 28S partial primer D1-D3. Maximum likelihood and Bayesian inference were performed for phylogenetic analysis. Morphometric and morphological data of the specimens studied ranged in accordance as observed in previous descriptions of the species. Observations from scanning electron microscopy and histology corroborated with those observed in stained whole mounts. Molecular analysis showed that specimens of P. bilabiatum from Brazil clustered with another two sequences of this species fromdifferent hosts and localities, with a high node support value. The integrative taxonomic approach allowed to record and describe new characteristics of P. bilabiatum related to the tegument, the structure and the arrangement of its tissues. The use of molecular markers confirmed that specimens identified as P. bilabiatum from different hosts and localities are all conspecific. Further studies, mainly molecular with less conserved genetic markers, should be carried out to better understand the phylogenetic relationships of Prosogonotrema with Hemiuroidea.

The geographic range of a species is crucial for obtaining information on the exact distribution of the species. The geographic data are important for delimiting distinct species or exploring the degree of differentiation among different populations of a species. The local details of species boundaries facilitate the study of the importance of phylogeographic background, secondary contacts, and hybrid zones, along with the relations between the species and its extrinsic environmental factors. In the present study, the range boundaries of Bufo bufo and Bufo verrucosissimus in the north-eastern region of Türkiye were delineated using an integrative taxonomic approach that utilized a combination of molecular and morphological data. According to the mtDNA results of the present study, B. bufo inhabits a single distribution from İyidere town to Çayeli town in Rize, while B. verrucosissimus is distributed from Şavşat town of Artvin to Ardeşen town in Rize. In addition, the two species coexist in Pazar, Hemşin, and Çamlıhemşin towns in Rize. The demographic analyses indicated a distinct population expansion for the B. verrucosissimus species after the Last Glacial Maximum, while the same did not occur for B. bufo. The univariate and multivariate statistical analyses conducted for the morphological data of the two species corroborated the presence of a putative contact zone between B. bufo and B. verrucosissimus. In summary, the present study resolved the non-distinct geographic boundaries between B. bufo and B. verrucosissimus species and also revealed the easternmost distribution of B. bufo in Türkiye. In addition, important evidence on the putative contact zone between the two species was indicated using an integrative taxonomic approach.

Clestobothrium Lühe, 1899 is a genus of cestodes belonging to the order Bothriocephalidea, which infects marine fish from the Gadiformes order. Herein, a novel species of Clestobothrium is described from the intestine of the European hake Merluccius merluccius (Linnaeus, 1758) in the Ionian Sea (Eastern Mediterranean Sea), through an integrative taxonomic approach that combines morphological and molecular data. The new species, named Clestobothrium brettiae n. sp., can be distinguished from all congeners by its unique ovary shape, and a combination of characters including the arrangement and number of testes. It shares similar morphological characters, with Clestobothrium crassiceps (Rudolphi, 1819) Lühe, 1899, which overlaps in host and geographic distribution. However, morphological differences between Clestobothrium brettiae n. sp. and C. crassiceps also include the arrangement of gladiate spinitriches, as well as a larger scolex, proglottids, testes and cirrus sac, and different ovary morphology. The establishment of Clestobothrium brettiae n. sp. as a novel taxon is supported by detailed morphological description and biometric statistics, in addition to molecular characterisation (based on partial small subunit ribosomal ribonucleic acid [rRNA], partial large subunit rRNA, and internal transcribed spacer region 2), genetic distance, and phylogenetic analyses.