This taxon has been reported from North America. [20] (Appendix 2: S2 and S3 Tables; available at: http://datadryad.org/resource/doi:10.5061/dryad.8rc4r). The results of the present analysis along with the analysis presented by Papakostas et al. We also provide in the supplementary materials an organized collection of ITS1 sequences available in GenBank that may be used to facilitate identification of each of these species using this particular molecular marker. Permanent glycerin glass slide mounts, each containing a single specimen, were prepared according to Jersabek et al. Once all trophi had been expelled from the lorica, they were pipetted between the remaining four 50 μL beads of distilled water. Brachionus plicatilis is a euryhaline (tolerate a wide range of salinity) rotifer in the Family Brachionidae, and is possibly the only commercially important rotifer, being raised in the aquaculture industry as food for fish larvae. This can be identified with species ‘C’ as suggested by the molecular analysis of this study and of Papakostas et al. calyciflorus) were obtained as 1.06 and 124.54 mg L-1, respectively (Table 1). Funding acquisition, nov. is characterized by the fact that the medial sinus is elevated with a well-marked median notch between short oval or nearly triangular protuberances. Investigation, [68], and deposited in the Frank J. Myers collection at the Academy of Natural Sciences in Philadelphia (ANSP). Formal analysis, urn:lsid:zoobank.org:act:0A4F5205-47ED-47ED-4C72-9686-0E33E0F34DA3. [49], and Michaloudi et al. [20] based on nuITSI sequence data: species ‘A’: B. dorcas; species ‘B’: B. elevatus sp. Brachionus calyciflorus Pallas, 1766: UKSI Synonym Source; Brachionis calyciflorus Pallas, 1766 synonym: UKSI Classification unranked Biota kingdom Animalia phylum Rotifera class Eurotatoria order Ploima family Brachionidae genus Brachionus species Brachionus calyciflorus. nov. e (range 29.54–74.2 μm, mean 49.82), r/b (range 0.164–0.424, mean 0.254) (S2 Table). All spines are triangular with a wide base and relatively sharp apices. A lot of the confusion undoubtedly arises from its high phenotypic variability [37]. ‘Nevada’ and B. here. Additional purging process of water vapour was applied during LTM to ensure higher relative humidity in a specimen chamber of the ESEM to prevent a sample collapse before its low temperature stabilisation. Trophi: Malleate type (Fig 7) bearing the characteristics of the genus as described by Segers et al. Further discrimination between B. dorcas and B. elevatus sp. Table 1: Acute 24 h toxicity of technical deltamethrin and imidacloprid insecticides in Brachionus calyciflorus. This latter character though exhibits a lot of variation (Fig 5). [20] and the present study, this hypothesis can be rejected. However, this has not prevented us from finding clear morphometric differences between the species. [20]. Photographs were taken by JSM-6300 Scanning Electron Microscope. No lateral notches exist. nov. compared to B. elevatus sp. Trophi: Malleate type (Fig 7) bearing the characteristics of the genus as described by Segers et al. B. calyciflorus distributing in the three lakes is a complex composed of two sibling species. By applying modern phylogenetic species delimitation methods (e.g., reviewed in Fontaneto et al., [23]), the species status of these cryptic species was further supported in a recent study [19]. A total of 136 individuals were examined coming from 10 clones established from resting eggs collected in 3 water bodies from the Netherlands (Table 1). Investigation, Since B. calyciflorus served as type species for the Brachionus genus [37], numerous morphological variants on subspecies and infrasubspecific level have been described (e.g., [38, 39]). Papakostas et al. Writing – original draft, The electronic edition of this article conforms to the requirements of the amended International Code of Zoological Nomenclature (ICZN), and hence the new names contained herein are available under that Code from the electronic edition of this article. Trophi: Malleate type (Fig 7) bearing the characteristics of the genus as described by Segers et al. Brachionus species Brachionus calyciflorus Name Synonyms Brachionis calyciflorus Pallas, 1766 Brachionus anuraeiformis Brehm, 1909 Brachionus calyciflorus dorcas Gosse, 1851 Brachionus calyciflorus f. anuraeiformis Brehm, 1909 Homonyms nov. seems to resemble the ‘c’ form they describe. Funding: Spiros Papakostas was supported by the Academy of Finland (grant number 258048). In the present study, the anteroventral structure with the marked protuberances of the medial sinus discriminated the individuals of the Brachionus elevatus from the individuals of the B. calyciflorus s.s. Kutikova & Fernando [39] based on this characteristic describe their ‘b’ form as being intermediate between Brachionus calyciflorus borgerti Apstein, 1907 and the typical form in the sense that the ‘b’ form lacks the saw-like basal tooth in the median spines of the anterodorsal margin of Brachionus calyciflorus borgerti although they have a broad base. No lateral notches exist. In addition to this, useful taxo- bioindicators of lake trophic state. Yes The variables weighing the most in this function were traits from the anteroventral margin, namely ‘q’, ‘v’, ‘r’ and ‘i’, traits representing overall body size (i.e. ... Walsh EJ, 2005. Copyright © 2014 Elsevier Inc. All rights reserved. We cordially thank Dr. Petr Jan Juračka for establishing the first contacts between some of the involved research groups and his initial help with the transportation of samples. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Nevertheless, its taxonomy remains particularly confused. Indeed, there is growing evidence that many rotifers feed on protozoans [for review see (Arndt, … Anterior dorsal margin with four spines, two on each side of a V-shaped sinus (Figs 4–6). For each case, when possible, we provide the specific name of the species, the country, and the area, lake or river in which it was recorded together with the reference source (Table A in S1 Text). Brachionus sp. To render species delimitations and descriptions more robust, molecular methods are nowadays increasingly combined with morphology, ecology, cross-fertilization data as well as with other sources of information (e.g., behavioural data, biogeography) in the framework of a taxonomical approach coined as integrative taxonomy [6, 7]. nov. e (range 34.62–72.31 μm), k (range 40.77–70.96 μm) and j (range 38.07–73.77 μm) (S2 Table). A total of 151 individuals were examined coming from 11 clones established from resting eggs collected in 5 water bodies in The Netherlands (Table 1). Yes The taxonomic identity of a name is nevertheless fundamental for nomenclatural stability [9, 10, 11]. It has a broad distribution in salt lakes around the world and has become a model system for studies in ecology and evolution https://doi.org/10.1371/journal.pone.0203168.g006. Competing interests: The authors have declared that no competing interests exist. Investigating how gene flow among species would affect morphometric traits may thus represent an intriguing topic of research to be done. Of all the material examined in the present study and Papakostas et al. https://doi.org/10.1371/journal.pone.0203168.g001. nov. are: B. dorcas e (range 43.84–92.62 μm, mean 67.16), r/b (range 0.102–0.253, mean 0.187), B. elevatus sp. Resources, Copyright © 2021 Elsevier B.V. or its licensors or contributors. nov. and B. fernandoi sp. We then measured several morphometric traits under standardized conditions and tested for the existence of morphometric differentiation among each of the four species pairs. Rami asymmetrical. Another point of consideration when interpreting our conclusions is related to the restricted geographic distribution of our studied samples. This situation has led [40] to express the need for a thorough revision of the taxon and the validation of these numerous variants recorded worldwide in ecological studies (Table A in S1 Text). Brachionus calyciflorus s.s. has a cosmopolitan distribution. Three antennae are present: one found in the U-shaped sinus between the medial anterodorsal spines when the coronal disc is extended, and two others on either lateral side of the lorica at the posterior part of the animal length. Brachionus calyciflorus is a species shape of dorsal projection over foot aperture. Institute of Scientific Instruments, Academy Of Sciences of the Czech Republic, Brno, Czech Republic, Roles [20]. For example, in the case of B. plicatilis Müller, 1786, morphological identifications based upon individuals raised under laboratory conditions [24] have proven to be inadequate for field studies in some cases [17, 77]. It comes form the Latin elevatus, past participle of elevare "lift up, raise". Project administration, Fly ash effluent decreases algal food level and quality in Lake Hui. This effort has resulted in the description of two new species, B. elevatus sp. The dimensions measured concerning these characteristics for B. dorcas and B. elevatus sp. Visualization, However, all investigated populations originated from a relatively restricted geographic area (i.e., the territory of The Netherlands). Additional tests showed significant differences among each pair of species with R2-values ranging between 19 and 55% (S1 Table). Two measurements of the anterodorsal side, namely ‘d’ and ‘f’, were not included in the further analysis due to high distortion of the placement of the anterior spines during preservation. Visualization, In Brachionus, ingestion is largely particle size dependent, where B. calyciflorus feeds most efficiently on particles with a diameter around 10 µm following an approximately bell-shaped feeding efficiency curve (Rothhaupt, 1990a, b). Brachionus is a typically species- rich and predominantly warm-water genus (Segers, 2008), and the geographical distributions of many species in this genus are certified to be limited to certain latitudinal ranges, predominantly tropical and subtropical (Segers and De Smet, 2008). In this study, we aim at comprehensively studying the morphometry and morphology of all four B. calyciflorus cryptic species recognized by Papakostas et al. Sibling species I distributes in Lake Hui through successful dispersal and colonization. [20]. Microphotographs for each individual were taken with an adjusted camera Canon Power shot A650 IS, and morphometric measurements were obtained using ImageJ [47]. [20]. ‘Grey’ literature (i.e. By combining phylogenetic, morphometric and morphological analyses, we confirm the existence of four cryptic species that have been recently suggested by a molecular study. nov. has a narrower anteroventral opening ‘i’, while B. calyciflorus s.s. has a wider anterodorsal side ‘b’. [48], Ciroz-Perez et al. Table 1: Acute 24 h toxicity of technical deltamethrin and imidacloprid insecticides in Brachionus calyciflorus. LC10 LC90 LC50 95% Confidence limits NOEC LOEC LC50 Deltamethrin (mg L-1) 0.70 0.80 1.06 1.02-1.11 0.81 1.40 Brachionus dorcas Gosse 1851 [70], p. 203, Brachionus calyciflorus dorcas, Gosse 1851 [70], p. 203, Brachionus dorcas spinosus, Wierzejski 1891 [71], p. 52, Fig 4, The name ‘dorcas’ comes from the Greek word ‘δορκάς’ which is an antelope. https://doi.org/10.1371/journal.pone.0203168.g004. The Brachionus calyciflorus PE500 library was downloaded from NCBI (SRA SRR6027265), and the same assembly and cleaning procedure was followed except that contigs with best blast hits only to bacteria in the blobtools pipeline were removed regardless of GC content or coverage. For the sake of continuity, we will henceforth use the term, haplotype, to describe unique sequences of ITS1. N2 - Chronic lead (Pb) toxicity tests with Brachionus calyciflorus, Chironomus tentans, and Lymnaea stagnalis were performed in artificial freshwaters. Shallow eutrophic city pond close to city center of The Hague (The Netherlands); N 52.090694°; E 4.338444°. Also, by combining a morphological analysis with an exhaustive literature survey, we were able to link the four species to previously described forms. A total of six individual rotifers were pipetted into the first 50 μL drop of distilled water, and 10 μL of domestic bleach (3% sodium hypochlorite solution) was added. Netherlands Institute of Ecology, Department of Aquatic Ecology, Wageningen, The Netherlands, Roles References Huang, L. "Effectof Aldrin on Life History Characteristics of Rotifer Brachionus calyciflorus Pallas." Sibling species II distributed in both Lake Tingtang and Lake Fengming, showing that its dispersal existed between the two lakes. Yes We gratefully acknowledge D. Fontaneto whose valuable suggestions were extremely helpful to finally shape the manuscript. Three antennae are present: one found in the V-shaped sinus between the medial anterodorsal spines when the coronal disc is extended, and two others on either lateral side of the lorica slightly posterior to the widest point of the lorica. No, PLOS is a nonprofit 501(c)(3) corporation, #C2354500, based in San Francisco, California, US, https://doi.org/10.1371/journal.pone.0203168, http://datadryad.org/resource/doi:10.5061/dryad.8rc4r, http://datadryad.org/review?doi=doi:10.5061/dryad.4ng70, - Wave-like anteroventral margin……………………………………………………2, - Median anterodorsal spines longer than lateral anterodorsal spines …………………………………………………………………………, - Width of the anteroventral medial sinus > 30 μm and more than 1/5 the lorica width (i.e. https://doi.org/10.1016/j.ecoenv.2014.09.036. It is found in freshwater habitats and it is an euplanktonic species. In the present investigation of Bhoj wetland, on an average basis, Brachionus caudatus (189 Ind.1-1) was recorded the maximum represented species density in the study period while (3.0 Brachionus urceus Manubria more similar in shape with B. dorcas, are triangular, pointed at the external sickle-shaped margin, flattened and bend at their distal end. As such, it provides a powerful tool to discover, describe and assign taxonomically valid names to previously unknown species and thus resolve many of the problems associated to taxonomic crypsis [8, 15]. Methodology, nov. (G, H). Based on the material analysed the present study confirms morphologically and genetically its Palearctic distribution. For this purpose, we collected and cultured additional rotifer clones representing multiple populations for each of the four suggested species. Subuncus brush-like. LC10 LC90 LC50 95% Confidence limits NOEC LOEC LC50 Deltamethrin (mg L-1) 0.70 0.80 1.06 1.02-1.11 0.81 1.40 Historically, these traits have been used to describe certain forms of Brachionus calyciflorus (i.e., B. calyciflorus f. anuraeiformis and B. calyciflorus f. amphiceros [38, 60, 72]). This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. For more information about PLOS Subject Areas, click Resources, This was the case for all three dimensions measured concerning this characteristic; B. dorcas: e (range 43.84–92.62 μm), k (range 52.41–86.21 μm) and j (range 46.32–98.38 μm), B. calyciflorus s.s. e (range 25.17–69.82 μm), k (range 31.14–68.89 μm) and j (range 25.15–74.05 μm), B. elevatus sp. Clonal lines were initially established by individually transferring hatched Brachionus calyciflorus females from the dishes into wells of a 24-well plate filled with chemostat grown Chlamydomonas reinhardtii (1000 μmol C L-1). The discriminating factor of the individuals of Brachionus dorcas is related to the anterodorsal spines, and specifically the fact that the median spines were much longer compared to the other groups. resting eggs were separated using the sugar flotation method [43]. Fulcrum short and hollow. Writing – original draft, nov. has no protrusion right before the median sinus of the anteroventral margin which is smooth with no lobes being formed by intermediate notches. Writing – original draft, nov. from B. calyciflorus s.s. is the opening of the median sinus of the anterovental margin which has the biggest opening; r range 21.33–45.50 B. fernandoi sp. Applying DFA on all four species revealed a particularly strong differentiation between species ‘A’ and the other three species, potentially overwhelming differences among the species ‘B’, ‘C’, and ‘D’; thus in order to discriminate the rest of the species as well DFA was applied on two different datasets: (a) including data of all four species and (b) including only the data of species ‘B’, ‘C’ and ‘D’. By applying an integrative taxonomy approach, Papakostas et al. Although this may limit the generality of the diagnostic traits suggested for species identification, this does not disqualify our conclusion that systematic differences between species exist. Fulcrum short and hollow.

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