Integrative Descriptions of Two New Tardigrade Species along with the New Record of Mesobiotus skorackii Kaczmarek et al., 2018 from Canada

Two new tardigrade species from a moss sample collected in Canada, one representing Macrobiotus hufelandi complex and the second one belonging to the genus Bryodelphax, are described. Integrative analysis was undertaken based on morphological and morphometric data (using both light and scanning electron microscopy (SEM)) combined with multilocus molecular analysis (nuclear sequences, i.e., 18S rRNA, 28S rRNA and ITS-2 as well as mitochondrial COI barcode sequences). Based on COI sequences, Macrobiotus birendrai sp. nov. is most similar to Mac. canaricus (p-distance 17%), whereas Bryodelphax mareki sp. nov. is most similar to Bry. parvulus (p-distance 16%). Both species differ also from their congeners in some morphological and morphometric characters of adults and/or details of egg chorion. Additionally, a large population of Mesobiotus skorackii was found in the sample and this is the first report of this species outside its terra typica in Kirghizia. The original description of this species was prepared based solely on the morphology and morphometry, therefore, here we provide updated data for this species enclosing morphometric and molecular data for the Canadian population.

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An integrative description of Ramazzottius subanomalus (Biserov, 1985 (Tardigrada) from Poland

Zootaxa ◽

10.11646/zootaxa.4300.3.4 ◽

2017 ◽

Vol 4300 (3) ◽

pp. 403 ◽

Cited By ~ 19

Author(s):

DANIEL STEC ◽

KRZYSZTOF ZAWIERUCHA ◽

ŁUKASZ MICHALCZYK

Keyword(s):

Dna Sequences ◽

Original Description ◽

Molecular Data ◽

28S Rrna ◽

Morphometric Traits ◽

Concrete Wall ◽

Morphometric Characters ◽

Mitochondrial Coi ◽

Qualitative And Quantitative ◽

Quantitative Characters

A population of Ramazzottius subanomalus (Biserov, 1985) was found in a moss sample collected from concrete wall in Poznań, western Poland. Animals were prepared for light and scanning electron microscopy and for DNA sequencing to provide an integrative description of the species that was originally described only by means of classical alpha taxonomy. As a result of our studies, we provide the first ever SEM photomicrographs of Ramazzottius subanomalus individuals and their buccal apparatuses. Additionally, we present new DNA sequences as well as new morphometric data for R. subanomalus. The molecular data comprise sequences for three DNA fragments, one mitochondrial (COI) and two nuclear (18S rRNA and 28S rRNA). As a result of being able to analyse a considerable number of animals and eggs, our study has expanded some of R. subanomalus morphometric traits ranges; compared with those provided in the original description. The spine-shaped egg processes as well as qualitative and quantitative characters of adults show R. subanomalus is most similar to Ramazzottius anomalus (Ramazzotti, 1962). However, our study has shown that R. subanomalus differs from R. anomalus by the lack of fine granulation on eggshell surface as well as by three other morphometric characters: longer buccal tube and two aspects of the placoids. We also discuss the validity of the R. anomalus record in Poland in the light of our findings. Finally, we show that the extreme eggshell variability in this species is observed both in natural and laboratory environments.

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Re-description of the Arctic tardigrade Tenuibiotus voronkovi (Tumanov, 2007 (Eutardigrada; Macrobiotidea), with the first molecular data for the genus

Zootaxa ◽

10.11646/zootaxa.4196.4.2 ◽

2016 ◽

Vol 4196 (4) ◽

pp. 498 ◽

Cited By ~ 8

Author(s):

KRZYSZTOF ZAWIERUCHA ◽

MAŁGORZATA KOLICKA ◽

ŁUKASZ KACZMAREK

Keyword(s):

Original Description ◽

Molecular Data ◽

Internal Transcribed Spacers ◽

The Arctic ◽

Morphological Description ◽

28S Rrna ◽

Morphometric Data ◽

Additional Material ◽

Svalbard Archipelago ◽

5.8S Rdna

Tardigrada is phylum of micrometazoans widely distributed throughout the world, because of old descriptions and insufficient morphometric data, many species currently need revision and re-description. Tenuibiotus voronkovi (Tumanov, 2007) is tardigrade previously only recorded from the Svalbard archipelago. This species’ original description was based on two individuals with destroyed claws on the fourth pair of legs and a lack of complete morphometric data for buccal tube and claws. In this paper, we present a re-description of T. voronkovi, supplementing the original description using the original paratype and additional material from Svalbard: Spitsbergen, Nordaustlandet and Edgeøya. This species is characterised by two macroplacoids and a microplacoid, claws of Tenuibiotus type, dentate lunules under claw IV, and faint granulation on legs I–III and strong granulation on the legs IV. We include a new morphological description with microphotographs, morphometric, and molecular data (including: mitochondrial cytochrome c oxidase subunit I (COI), internal transcribed spacers (ITS1–5.8S rDNA–ITS2), and nuclear ribosome subunits 28S rRNA and 18S rRNA). These are the first published molecular data for the genus Tenuibiotus Pilato and Lisi, 2011, analysis of which indicated an affiliation of Tenuibiotus to the family Macrobiotidae. We found no differences in body size between individuals from different islands (Nordaustlandet and Edgeøya), but did observe variability in the eggs. After revision of the literature and the published figures, we concluded that Dastych’s (1985) report of T. willardi (Pilato, 1976) from Svalbard, was actually T. voronkovi, which has the greater distribution in Svalbard, and other Arctic locations, than previously believed.

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Taxonomical considerations and molecular phylogeny of the closely related genera Bitylenchus, Sauertylenchus and Tylenchorhynchus (Nematoda: Telotylenchinae), with one new and four known species from Iran

Journal of Helminthology ◽

10.1017/s0022149x20000784 ◽

2020 ◽

Vol 94 ◽

Author(s):

M. Hosseinvand ◽

A. Eskandari ◽

S. Ganjkhanloo ◽

R. Ghaderi ◽

P. Castillo ◽

...

Keyword(s):

New Species ◽

Anterior Part ◽

Phylogenetic Analyses ◽

Ribosomal Subunit ◽

Molecular Data ◽

28S Rrna ◽

Natural Habitats ◽

Morphometric Characters ◽

Molecular Approaches ◽

Tail Tip

Abstract During several nematological surveys in cultivated and natural habitats in Khuzestan and Zanjan provinces of Iran, a new species, Bitylenchus parvulus n. sp., two new records for Iran – namely, Tylenchorhynchus agri and Tylenchorhynchus graciliformis – and a population of Bitylenchus parvus and one of Sauertylenchus maximus were recovered and characterized based upon morphological and molecular approaches. The new species is characterized by lip region with five to seven annuli, stylet 17.7 (17.0–18.5) μm long, sub-cylindrical tail narrowing abruptly near terminus giving a bluntly digitate shape to the tail tip, cuticle near anterior part of vulva wrinkled and post-rectal sac occupies whole of tail cavity. The phylogenetic analyses were carried out using molecular data from D2–D3 expansion segments of large ribosomal subunit (28S rRNA) for all studied species and the partial small ribosomal subunit (18S rRNA) for the new species. The representatives of Bitylenchus and Sauertylenchus formed distinct clades from Tylenchorhynchus members, supporting the hypothesis in which Bitylenchus and Sauertylenchus could be considered as valid genera, but rejecting the ‘large-genus’ concept for Tylenchorhynchus. Also, Sauertylenchus ibericus was proposed as a junior synonym of S. maximus based on the results from morphological and phylogenetic analysis. Furthermore, an identification key for all known species included in the three genera Bitylenchus, Tylenchorhynchus and Sauertylenchus is presented herein. The number of transverse annuli on the lip region and presence/absence of post-rectal sac were considered as the main diagnostic characters for classifying the species into seven groups, and other morphological and morphometric characters were subsequently used for distinguishing species in each group.

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An example of problems associated with DNA barcoding in tardigrades: a novel method for obtaining voucher specimens

Zootaxa ◽

10.11646/zootaxa.3104.1.3 ◽

2011 ◽

Vol 3104 (1) ◽

pp. 42 ◽

Cited By ~ 15

Author(s):

MICHELE CESARI ◽

ILARIA GIOVANNINI ◽

ROBERTO BERTOLANI ◽

LORENA REBECCHI

Keyword(s):

Dna Barcoding ◽

Dna Barcode ◽

Intraspecific Variability ◽

Original Description ◽

Morphological Characters ◽

Molecular Data ◽

Integrative Taxonomy ◽

Type Locality ◽

Moss Sample ◽

Voucher Specimens

We have in recent papers revealed that an integrative taxonomy approach helps to solve taxonomic problems in tardigrades. However, whole tardigrades are required for DNA work, which leaves no hologenophore voucher specimens with adult morphology. Using a novel methodology for the Tardigrada, we introduce the practice of collecting high quality maximum magnification light microscopy images of recently thawed animals to act as hologenophore voucher specimens of animals later used for DNA barcode sequencing. Within the framework of a DNA barcoding project on tardigrades, we collected a moss sample from the type locality of Macrobiotus terminalis Bertolani & Rebecchi, 1993 (Castelsantangelo, Central Apennines, Italy), a species of the “Macrobiotus hufelandi group”. Within the moss sample we found several animals and eggs with a morphology that corresponded to the original description of M. terminalis, while others were attributable to Macrobiotus macrocalix Bertolani & Rebecchi, 1993. In this study, molecular (cox1 mtDNA) analyses demonstrated no intraspecific variability in M. terminalis from the type locality but very large interspecific differences when compared with M. macrocalix and GenBank data for other species within the M. “hufelandi group”. There was also a large difference between our M. terminalis sequences and the GenBank data of a specimen attributed to the same species. The GenBank sequence originated from a population in the Northern Apennines, whose morphology appeared to be like that of the specimens of the locus typicus. This confirmed the importance in utilising material from the type locality for linking molecular data to the species’ morphological characters. Our paper underlines the importance of an integrative taxonomy in species diagnoses and demonstrates a scenario where morphological observations alone are not always sufficient. Lastly, this work adds reliable information to the sequence reference library that provides a useful building block for further studies on similar and related tardigrade taxa.

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Integrative descriptions of two new Macrobiotus species (Tardigrada, Eutardigrada, Macrobiotidae) from Mississippi (USA) and Crete (Greece)

Zoosystematics and Evolution ◽

10.3897/zse.97.65280 ◽

2021 ◽

Vol 97 (1) ◽

pp. 281-306

Author(s):

Matteo Vecchi ◽

Daniel Stec

Keyword(s):

18S Rrna ◽

Molecular Data ◽

Integrative Taxonomy ◽

Distal Portion ◽

Morphological Data ◽

28S Rrna ◽

Morphometric Characters ◽

Clade B ◽

Specific Morphology ◽

Definition Of

In this paper, we describe two new Macrobiotus species from Mississippi (USA) and Crete (Greece) by means of integrative taxonomy. Detailed morphological data from light and scanning electron microscopy, as well as molecular data (sequences of four genetic markers: 18S rRNA, 28S rRNA, ITS-2 and COI), are provided in support of the descriptions of the new species. Macrobiotus annewintersaesp. nov. from Mississippi belongs to the Macrobiotus persimilis complex (Macrobiotus clade B) and exhibits a unique egg processes morphology, similar only to Macrobiotus anemone Meyer, Domingue & Hinton, 2014, but mainly differs from that species by the presence of eyes, granulation on all legs, dentate lunulae on legs IV, and of bubble-like structures within the tentacular arms that are present on the distal portion of the egg processes. Macrobiotus rybakisp. nov. from Crete belongs to the Macrobiotus clade A and is most similar to Macrobiotus dariae Pilato & Bertolani, 2004, Macrobiotus noemiae Roszkowska & Kaczmarek, 2019, Macrobiotus santoroi Pilato & D’Urso, 1976, and Macrobiotus serratus Bertolani, Guidi & Rebecchi, 1996, but differs from them mainly in the morphological details of its egg processes and chorion reticulation, but also by a number of morphometric characters. In light of the specific morphology of the egg processes of Macrobiotus annewintersaesp. nov. and Macrobiotus anemone, that are equipped with tentacular arms instead of proper terminal disc, we also provide an updated definition of the Macrobiotus persimilis complex.

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Toward a revision of the genus Chelidura Latreille, 1825: designation of the Neotype for Chelidura aptera (Megerle in Charpentier, 1825) (Dermaptera: Forficulidae)

Memorie della Società Entomologica Italiana ◽

10.4081/memoriesei.2020.279 ◽

2021 ◽

pp. 279-302

Author(s):

Paolo Fontana ◽

Federico Pedrazzoli ◽

Valeria Malagnini ◽

Enrico Ruzzier ◽

Federico Marangoni ◽

...

Keyword(s):

Male Genitalia ◽

Type Species ◽

Original Description ◽

Molecular Data ◽

Type Material ◽

Dna Barcodes ◽

Massif Central ◽

Coi Sequences ◽

Morphological Investigations

Since it is not possible, after several years of research, to locate the type material of Chelidura aptera (Megerle in Charpentier, 1825), type species of the genus Chelidura latreille, 1825, and not being able to define in any way, even vaguely, the terra typica, it was decided to fix a neotype. This neotype is here selected on the basis of its matching with the original description and with the support of molecular data. Three Alpine populations, two Italian and one Swiss, of what is actually considered C. aptera were sampled and sequenced; COI sequences were then compared with Chelidura pyrenaica arverna David & Van Herrewege, 1973 from the Massif Central in France, and other species of the genera Chelidurella, Mesochelidura and Forficula. The separation between C. aptera and C. pyrenaica is strongly supported by DNA barcodes and substantial differences among the three Alpine populations have been detected. The genetically intermediate among the Alpine populations (Italy, Piedmont (Biella), Pennine Alps, lago del Mucrone) is chosen as neotype of Chelidura aptera, furthermore largely matching the original description. Further genetic and morphological investigations will clarify the relations among the Alpine populations and in the early future we expect to extend the analyzes to the majority of the Alpine and non-Alpine populations of Chelidura. In addition, the updated view about the taxonomy and distribution of Chelidura and the first illustration of male genitalia of Chelidura pyrenaica arverna David & Van Herrewege, 1973 are provided.

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Integrative description of a new Tunisian tardigrade species, Macrobiotus azzunae sp. nov. (Eutardigrada, Macrobiotidae, hufelandi group)

European Journal of Taxonomy ◽

10.5852/ejt.2021.758.1429 ◽

2021 ◽

Vol 758 ◽

Author(s):

Jamila Ben Marnissi ◽

Michele Cesari ◽

Lorena Rebecchi ◽

Roberto Bertolani

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

18S Rrna ◽

Integrated Approach ◽

Molecular Data ◽

28S Rrna ◽

Considerable Potential ◽

Mitochondrial Coi ◽

Taxonomic Approach ◽

Scanning Electron

In this paper a new tardigrade species, Macrobiotus azzunae sp. nov., from Tunisia, is described. An integrative taxonomic approach was applied by combining morphological, morphometric and molecular data. In particular, light and scanning electron microscopy observations, and four genetic markers, three nuclear (18S rRNA, 28S rRNA and ITS-2) and one mitochondrial (COI) were used. The analysis showed that M. azzunae sp. nov. belongs to the Macrobiotus hufelandi group and is most similar to Macrobiotus sandrae Bertolani & Rebecchi, 1993. It differs from M. sandrae by a more pronounced constriction of the first macroplacoid (hardly visible in M. sandrae) and for the eggshell shape, with thinner wires of the reticulum and meshes around the processes larger than the inter-process meshes in M. azzunae sp. nov., while all meshes are similar in size in M. sandrae. The species is gonochoristic. With this discovery, there are 33 species of tardigrades identified in Tunisia, all non-marine. This result, compared with nearby Sicily, where more research has been conducted, indicates that there is a considerable potential for identification of new species. Further research will be most informative if multiple habitats are explored and if carried out with an integrated approach as done in this present work.

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Remarkable Cryptic Diversity of Paratylenchus spp. (Nematoda: Tylenchulidae) in Spain

Animals ◽

10.3390/ani11041161 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1161

Author(s):

Ilenia Clavero-Camacho ◽

Carolina Cantalapiedra-Navarrete ◽

Antonio Archidona-Yuste ◽

Pablo Castillo ◽

Juan Emilio Palomares-Rius

Keyword(s):

Molecular Data ◽

Parasitic Nematodes ◽

28S Rrna ◽

Cryptic Diversity ◽

Undescribed Species ◽

Plant Parasitic Nematodes ◽

Mitochondrial Coi ◽

Taxonomic Approach ◽

Morphological And Molecular Data ◽

Plant Parasitic

In previous studies, fifteen species of Paratylenchus, commonly known as pin nematodes, have been reported in Spain. These plant-parasitic nematodes are ectoparasites with a wide host range and global distribution. In this research, 27 populations from twelve Paratylenchus species from 18 municipalities in Spain were studied using morphological, morphometrical and molecular data. This integrative taxonomic approach allowed the identification of twelve species, four of them were considered new undescribed species and eight were already known described. The new species described here are P. caravaquenus sp. nov., P. indalus sp. nov., P. pedrami sp. nov. and P. zurgenerus sp. nov. As for the already known described species, five were considered as first reports for the country, specifically P.enigmaticus, P. hamatus, P. holdemani, P. israelensis, and P. veruculatus, while P. baldaccii, P. goodeyi and P. tenuicaudatus had already been recorded in Spain. This study provides detail morphological and molecular data, including the D2-D3 expansion segments of 28S rRNA, ITS rRNA, and partial mitochondrial COI regions for the identification of different Paratylenchus species found in Spain. These results confirm the extraordinary cryptic diversity in Spain and with examples of morphostatic speciation within the genus Paratylenchus.

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Macrobiotus polypiformis sp. nov., a new tardigrade (Macrobiotidae; hufelandi group) from the Ecuadorian Pacific coast, with remarks on the claw abnormalities in eutardigrades

European Journal of Taxonomy ◽

10.5852/ejt.2017.327 ◽

2017 ◽

Cited By ~ 5

Author(s):

Milena Roszkowska ◽

Marta Ostrowska ◽

Daniel Stec ◽

Karel Janko ◽

Łukasz Kaczmarek

Keyword(s):

New Species ◽

Taxonomic Status ◽

28S Rrna ◽

Microscopy Imaging ◽

Morphometric Characters ◽

Mitochondrial Coi ◽

Taxonomic Description ◽

As Species ◽

Species Inquirenda

From a moss sample collected in the Manabí Province in Ecuador, we extracted 96 specimens of a new species of eutardigrade. No eggs were found. In order to obtain eggs, an in vitro culture was prepared. In total, 136 specimens (including ten simplex), one exuvia and 44 eggs (including two with embryos) of the new species were obtained. In addition to the traditional taxonomic description with morphometrics, light and scanning microscopy imaging, we also provide nucleotide sequences of three nuclear (18S rRNA, 28S rRNA, ITS-2) and one mitochondrial (COI) DNA fragments of the new species. Macrobiotus polypiformis sp. nov. belongs to the hufelandi group and is most similar to Ma. paulinae Stec, Smolak, Kaczmarek & Michalczyk, 2015, but differs from it mainly by the lack of dorso-lateral patches of granulation on the cuticle, egg processes with longer and more numerous filaments and in some morphometric characters of both eggs and adults. Moreover, we provide a short discussion on the modifications/abnormalities of the claws in eutardigrades and the possible consequences on the taxonomic status of Mesobiotus armatus (Pilato & Binda, 1996), suggesting its consideration as species inquirenda (with uncertain taxonomic status).

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Molecular characterisation and phylogenetic position of Hemicycliophora labiata Colbran, 1960, from turfgrass in Korea, with comments on its morphology

Nematology ◽

10.1163/15685411-bja10002 ◽

2020 ◽

Vol 22 (8) ◽

pp. 927-938

Author(s):

Abraham Okki Mwamula ◽

Yeong Ho Kim ◽

Heebeen Na ◽

Hyeon Jeong An ◽

Young Ho Kim ◽

...

Keyword(s):

Poa Pratensis ◽

Morphological Characteristics ◽

Morphological Plasticity ◽

Original Description ◽

Molecular Data ◽

Integrative Approach ◽

Phylogenetic Position ◽

28S Rrna ◽

Molecular Characterisation ◽

Species Complexes

Summary Hemicycliophora labiata was reported from the rhizosphere of Poa pratensis and for the first time in Korea. Females and juveniles are molecularly characterised and morphological and morphometric descriptions supplied. Species identification was made using an integrative approach considering morphological characteristics and the results of the analyses of the D2-D3 expansion segment of 28S rRNA, ITS1-5.8S-ITS2 of rRNA and the partial COI gene sequences. The phylogenetic position of H. labiata is resolved after recent misidentifications due to the existence of a wide morphological plasticity within the genus. Females and juveniles from Korea conform to the original description and also to subsequent species descriptions from Australia, South Africa, Sri Lanka, and India. Despite the close morphological similarities with H. typica de Man, 1921, the two species can be adequately differentiated based on molecular data. This is the first molecular characterisation of H. labiata. The use of molecular markers in species delimitation continues to explore the existence of species complexes, thus providing a better understanding of the unresolved species biodiversity within this speciose genus.

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