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Revista mexicana de biodiversidad ISSN: 1870-3453 ISSN: 2007-8706 Instituto de Biología Lagunas-Calvo, Omar; Hernández-Mena, David Iván; Rivas, Gerardo; Contreras-Mirón, Samantha; Mendoza-Garfias, Berenit; Echeverría-Martínez, Olga M.; Oceguera-Figueroa, Alejandro Rediscovery of the nemertean Prostoma graecense from Xochimilco, 75 years after its first and only record Revista mexicana de biodiversidad, vol. 89, no. 1, 2018, pp. 39-45 Instituto de Biología DOI: 10.22201/ib.20078706e.2018.1.2094 Available in: http://www.redalyc.org/articulo.oa?id=42559253005 How to cite Complete issue Scientific Information System Redalyc More information about this article Network of Scientific Journals from Latin America and the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Project academic non-profit, developed under the open access initiative Revista Mexicana de Biodiversidad 89 (2018): 39-45 Taxonomy and systematics Rediscovery of the nemertean Prostoma graecense from Xochimilco, 75 years after its first and only record Redescubrimiento del nemertino Prostoma graecense de Xochimilco, 75 años después de su primer y único registro Omar Lagunas-Calvo a, David Iván Hernández-Mena b, Gerardo Rivas a, Samantha Contreras- Mirón b, Berenit Mendoza-Garfias c, Olga M. Echeverría-Martínez d, Alejandro Oceguera- Figueroa b, * a Departamento de Biología Comparada, Facultad de Ciencias, Universidad Nacional Autónoma de México, Apartado Postal 70-399, 04510 México City, Mexico b Laboratorio de Helmintología, Instituto de Biología, Universidad Nacional Autónoma de México, Apartado postal 70-153, 04510 México City, Mexico c Laboratorio de Microscopia Electrónica, Instituto de Biología, Universidad Nacional Autónoma de México, Apartado postal 70-153, 04510 México City, Mexico d Laboratorio de Microscopia Electrónica, Facultad de Ciencias, Universidad Nacional Autónoma de México, Apartado Postal 70-181, 04510 México City, Mexico *Corresponding author: [email protected] (A. Oceguera-Figueroa). Recibido: 23 enero 2017; aceptado: 27 septiembre 2017 Abstract The identity of the freshwater nemertean Prostoma graecense (Böhming, 1892) from Lake Xochimilco is solved 75 years after its first recording. Identification of the specimens was accomplished via both morphological and molecular means, the latter employing the mitochondrial cytochrome c oxidase subunit I. Recent literature and our results suggest that this species has a cosmopolitan distribution and has, presumably, been recently introduced to areas well removed from its original distribution, which is unknown. The current study provides a background for continuing the work with freshwater and marine nemerteans and underscores the importance of including molecular data in future studies of the group. Keywords: Freshwater invertebrates; Nemertea; Tetrastemmatidae; Cox1; Taxonomy Resumen Se resuelve la identidad taxonómica del nemertino dulceacuícola Prostoma graecense (Böhming, 1892) del lago de Xochimilco 75 años después del primer registro. La identificación de los ejemplares se realizó con base en caracteres morfológicos y moleculares, esta última empleando el citocromo c oxidasa subunidad I mitocondrial. La literatura reciente y nuestros resultados sugieren que esta especie tiene una distribución cosmopolita y que ha sido introducida, presumiblemente, en áreas lejanas de su distribución original, la cual es desconocida. El presente estudio proporciona un contexto para continuar los estudios sobre nemertinos de agua marina y dulceacuícola y enfatiza la importancia de incluir datos moleculares en futuros estudios del grupo. Palabras clave: Invertebrados dulceacuícolas; Nemertea; Tetrastemmatidae; Cox1, Taxonomía ISSN versión impresa: 1870-3453; versión electrónica: 2007-8706; Universidad Nacional Autónoma de México, Instituto de Biología. Open Access bajo la licencia CC BY-NC-ND (4.0) https://doi.org/10.22201/ib.20078706e.2018.1.2094 40 O. Lagunas-Calvo et al. / Revista Mexicana de Biodiversidad (2018): 39-45 https://doi.org/10.22201/ib.20078706e.2018.1.2094 Introduction 1758, tilapia Oreochromis niloticus (Linnaeus, 1758), and the water hyacinth Eichhornia crassipes (Mart.) Solms. Members of the phylum Nemertea (ribbon worms) are (Cervantes-Sánchez & Rojas-Rabiela, 2000; Zambrano et characterized by the presence of an eversible muscular al., 2010). The aim of the present study is to document the proboscis housed within a cavity called a rhynchocoel; this presence of the freshwater nemertean in Xochimilco, an structure is found in no other animal group. Nemerteans area that has been severely impacted by human activities. are unsegmented, bilaterian, vermiform animals with a In addition, this study aims to solidify the taxonomic and closed circulatory system and protonephridia as excretory nomenclatural status of the nemertean and to generate organs. The body size of these worms range in length from the mitochondrial cytochrome c oxidase subunit I (cox1) a few millimeters up to 30 meters (Sundberg & Strand, DNA sequence, the preferred zoological barcoding region, 2010; Strand & Sundberg, 2015), representing the longest which may serve as a framework for future studies of this metazoan on Earth. Members of Nemertea are mostly charismatic taxon. marine, with only 22 out of the 1,280 described species inhabiting freshwater (Kajihara et al., 2008; Sundberg & Materials and methods Gibson, 2008; Strand & Sundberg, 2015). Most of the species diversity within the group occurs in the Nearctic In February, April, and October of 2016, 17 nemerteans and Palearctic, but this may only reflect the intensity of were collected from “Embarcadero de Cuemanco”, sampling efforts and taxonomic work rather than a real Xochimilco, México City, Mexico (19º17’20.934” N, biogeographical pattern (Strand & Sundberg, 2015). Because 99º06’06.585” W). Specimens were found within the roots of the difficulty of identifying preserved specimens, several of water lily Eichhornia crassipes that were collected from authors have suggested that morphological identification the edges of canals. The aquatic plants were placed in should always be accompanied by DNA sequence analyses trays and, after about 10 minutes, ribbon worms were (Chen et al., 2010; Sundberg & Strand, 2010; Sundberg et recovered from the bottom of the tray with the use of al., 2010, 2016). delicate paintbrushes. For subsequent morphological Only a single freshwater nemertean is known from procedures, worms were relaxed in 7% MgCl2 and fixed Mexico, originally referred to as Stichostemma rubrum in 4% formaldehyde. Two specimens were stained with (Leydi, 1850) from Lake Xochimilco, México City (Rioja, Gomori trichrome and mounted on permanent slides with 1941). Gibson and Moore (1976) listed S. rubrum under Canada balsam. For histological procedures, worms were the genus Prostoma and concluded that no typically North fixed in 10% formaldehyde in phosphate-buffered saline American species of Prostoma can be recognized, thus (PBS) at pH 7.2, dehydrated with graded ethanol series and the Mexican records of P. rubrum should be considered embedded in Epon epoxy resin. Semi-thin serial sections, as P. graecense. In the most recent taxonomic account of obtained with a Leica Ultracut UTC Ultramicrotome, were freshwater nemerteans from North America, only 2 species stained with toluidine blue. In addition, 2 worms were were recognized: Prostoma canadiensis Gibson & Moore, dehydrated and critical point dried with CO2, mounted and 1978, which possesses a frontal organ (including previous coated with a mixture of gold-palladium and subsequently records of P. graecense), and Prostoma asensoriatum observed in a Hitachi model SUI510 scanning electron (Montgomery, 1896), which lacks such an organ (Strand & microscope (SEM) at the Laboratorio Nacional de Sundberg, 2015). To robustly infer the presence of a frontal Biodiversidad, Instituto de Biología, Universidad Nacional organ, histological preparations are required, a procedure Autónoma de México (IB-UNAM). Voucher specimens that has yet to be used to characterize nemerteans from were deposited in the National Invertebrates Collection Mexico. at the IB-UNAM (accession number CNINV-002, 003). More than 75 years have passed since the first and only For molecular analyses, cox1 sequences from 3 specimens record of the freshwater nemertean from Xochimilco, an were obtained via procedures described elsewhere (Kvist area well known for its expansive biodiversity and number et al., 2014; Oceguera-Figueroa et al., 2005). The newly of endemics; arguably the most charismatic of these is generated cox1 sequences were aligned and compared with Axolotl [Ambystoma mexicanum (Shaw & Nodder, 1798)] 15 sequences of members of the suborder Monostilifiera and the Montezuma leopard frog [Lithobates montezumae (Class Enopla) available from GenBank (Table 1) and (Baird, 1854)]. During that time, Xochimilco has changed selected based on BLASTn (Altschul et al., 1990) matches. dramatically, due to a steep increase in human population Nucleotide sequences were aligned with ClustalW numbers and the intentional and unintentional introduction (Thompson et al., 1997), implemented in the web-version of of several exotic species that have reshaped the ecosystem, the software at http://www.genome.jp/tools/clustalw/. The including the common carp Cyprinus carpio Linnaeus, edges of the alignment were trimmed to include sequences M. M. Contreras-Pacheco et al. / Revista Mexicana de Biodiversidad 89 (2018): 39-45 41 https://doi.org/10.22201/ib.20078706e.2018.1.2094