Integrative taxonomy reveals two new cryptic species of Hyphessobrycon Durbin, 1908 (Teleostei: Characidae) from the Maracaçumé and middle Tocantins River basins, Eastern Amazon region
Abstract
Two new species, Hyphessobrycon frickei Guimarães, Brito, Bragança, Katz & Ottoni sp. nov. and H. geryi Guimarães, Brito, Bragança, Katz & Ottoni sp. nov., are herein described, based on seven different and independent species delimitation methods, and on molecular and morphological characters, making the hypothesis of these new species supported from an integrative taxonomy perspective. They belong to the “Rosy tetra” clade, which is mainly characterized by the presence of a dark brown or black blotch on the dorsal fin and the absence of a midlateral stripe on the body. These two new species are distinguished from the other members of this clade mainly by the arrangement, shape and color pattern of humeral and dorsal-fin spots, as well as by other characters related to scale counts and body pigmentation. The placement of the new species within the “Rosy tetra” clade was based on the combination of morphological character states mentioned above and corroborated by a molecular phylogenetic analysis using the mitochondrial gene cytochrome oxidase subunit 1. In addition, a new clade (here termed Hyphessobrycon copelandi clade) within the “Rosy tetra” clade is proposed based on molecular data, comprising H. copelandi, H. frickei sp. nov., H. geryi sp. nov. and a still undescribed species. Our results corroborate the occurrence of hidden species within the “Rosy tetra” clade, as suggested by previous studies.
References
Alfaro M.E. & Holder M.T. 2006. The posterior and the prior in Bayesian phylogenetics. Annual Review of Ecology, Evolution, and Systematics 37: 19–42. https://doi.org/10.1146/annurev.ecolsys.37.091305.110021
Amorim F.P. 2018. Jenynsia lineata species complex, revision and new species description (Cyprinodontiformes: Anablepidae). Journal of Fish Biology 92: 1312–1332. https://doi.org/10.1111/jfb.13587
Avise J.C. 2000. Phylogeography: the History on Formation of Species. University Press, Cambridge.
Benine R.C., Melo B.F., Castro R.M.C. & Oliveira C. 2015. Taxonomic revision and molecular phylogeny of Gymnocorymbus Eigenmann, 1908 (Teleostei, Characiformes, Characidae). Zootaxa 3956 (1): 1–28. https://doi.org/10.11646/zootaxa.3956.1.1
Betancur-R R., Arcila D., Vari R.P., Hughes L.C., Oliveira C., Sabaj M.H. & Ortí G. 2018. Phylogenomic incongruence, hypothesis testing, and taxonomic sampling: The monophyly of characiform fishes. Evolution 73 (2): 329–345. https://doi.org/10.1111/evo.13649
Benzaquem D.C., Oliveira C., da Silva Batista J., Zuanon J. & Porto J.I.R. 2015. DNA Barcoding in Pencilfishes (Lebiasinidae: Nannostomus) Reveals Cryptic Diversity across the Brazilian Amazon. PLoS ONE 10 (2): e0112217. https://doi.org/10.1371/journal.pone.0112217
Bickford D., Lohman D.J., Sodhi N.S., Ng P.K.L., Meier R., Winker K., Ingram K.K. & Das I. 2006. Cryptic species as a window on diversity and conservation. Trends in Ecology and Evolution 22 (3): 148–155. https://doi.org/10.1016/j.tree.2006.11.004
Bittencourt P.S., Campos Z., Muniz F.L., Marioni B., Souza B.C., Da Silveira R., de Thoisy B., Hrbek T. & Farias I.P. 2019. Evidence of cryptic lineages within a small South American crocodilian: the Schneider’s dwarf caiman Paleosuchus trigonatus (Alligatoridae: Caimaninae). PeerJ 7: e6580. https://doi.org/10.7717/peerj.6580
Bragança P.H.N., Ottoni F.P. & Rangel-Pereira F.S. 2015. Hyphessobrycon ellisae, a new species from northeastern Brazil (Teleostei: Characidae). Ichthyological Exploration of Freshwaters 26 (3): 255–262.
de Brito P.S., Guimarães E.C., Carvalho-Costa L.F. & Ottoni F.P. 2019. A new species of Aphyocharax Günther, 1868 (Characiformes, Characidae) from the Maracaçumé river basin, eastern Amazon. Zoosystematics and Evolution 95 (2): 507–516. https://doi.org/10.3897/zse.95.36788
Carvalho F.R. & Malabarba L.R. 2015. Redescription and osteology of Hyphessobrycon compressus (Meek) (Teleostei: Characidae), type species of the genus. Neotropical Ichthyology 13 (3): 513–540. https://doi.org/10.1590/1982-0224-20140173
Carvalho M.L., Costa Silva G.J,. Melo S., Ashikaga F.Y., Shimabukuro-Dias C.K., Scacchetti P.C., Devidé R., Foresti F. & Oliveira C. 2018. The non-monotypic status of the neotropical fish genus Hemiodontichthys (Siluriformes, Loricariidae) evidenced by genetic approaches. Mitochondrial DNA Part A 29 (8): 1224–1230. https://doi.org/10.1080/24701394.2018.1431230
Castro-Paz F.P., Batista J.S. & Porto J.I.R. 2014. DNA Barcodes of Rosy Tetras and Allied Species (Characiformes: Characidae: Hyphessobrycon) from the Brazilian Amazon Basin. PLoS ONE 9 (5): e98603. https://doi.org/10.1371/journal.pone.0098603
Chenna R., Sugawara H., Koike T., Lopez R., Gibson T.J., Higgins D.G. & Thompson J.D. 2003. Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Research 31: 3497–3500. https://doi.org/10.1093/nar/gkg500
Close B., Banister K., Baumans V., Bernoth E.M., Bromage N., Bunyan J., Erhardt W., Flecknell P., Gregory N., Hackbarth H. & Morton D. 1996. Recommendations for euthanasia of experimental animals: Part 1. Laboratory Animals 30 (4): 293e316. https://doi.org/10.1258/002367796780739871
Close, B., Banister, K., Baumans, V., Bernoth, E.M., Bromage, N., Bunyan, J., Erhardt, W., Flecknell, P., Gregory, N., Hackbarth, H. & Morton, D. 1997. Recommendations for euthanasia of experimental animals: Part 2.Laboratory Animals 31 (1): 1e32. https://doi.org/10.1258/002367797780600297
Costa W.J.E.M. & Amorim P.F. 2011. A new annual killifish species of the Hypsolebias flavicaudatus complex from the São Francisco River basin, Brazilian Caatinga (Cyprinodontiformes: Rivulidae). Vertebrate Zoology 61 (1): 99–104.
Costa W.J.E.M., Amorim P.F. & Mattos J.L.O. 2012. Species delimitation in annual killifishes from the Brazilian Caatinga, the Hypsolebias flavicaudatus complex (Cyprinodontiformes: Rivulidae): implications for taxonomy and conservation. Systematics and Biodiversity 10: 71–91. https://doi.org/10.1080/14772000.2012.664177
Costa W.J.E.M., Amorim P.F. & Aranha G.N. 2014. Species limits and DNA barcodes in Nematolebias, a genus of seasonal killifishes threatened with extinction from the Atlantic Forest of south-eastern Brazil, with description of a new species (Teleostei: Rivulidae). Ichthyological Exploration of Freshwaters 24 (3): 225–236.
Costa W.J.E.M., Cheffe M.M. & Amorim P.F. 2017. Two new seasonal killifishes of the Austrolebias adloffi group from the Lagoa dos Patos basin, southern Brazil (Cyprinodontiformes: Aplocheilidae). Vertebrate Zoology 67 (2): 139–149.
Darriba D., Taboada G.L., Doallo R. & Posada D. 2012. jModelTest 2: more models, new heuristics and parallel computing. Nature Method 9 (8): 772. https://doi.org/10.1038/nmeth.2109
Davis J.I. & Nixon K.C. 1992. Populations, genetic variation, and the delimitation of phylogenetics species. Systematic Biology 41 (4): 421–435. https://doi.org/10.1093/sysbio/41.4.421
Dayrat B. 2005. Towards integrative taxonomy. Biological Journal of the Linnean Society 85 (3): 407–415. https://doi.org/10.1111/j.1095-8312.2005.00503.x
Desalle R., Egan M.G. & Siddall M. 2005. The unholy trinity: taxonomy, species delimitation and DNA barcoding. Philophical Transactions of the Royal Society B 360: 1905–1916. https://doi.org/10.1098/rstb.2005.1722
Drummond A.J., Suchard M.A., Xie D. & Rambaut A. 2012. Bayesian phylogenetics with BEAUti and the BEAST 1.7. Molecular Biology and Evolution 29: 1969–1973.
Eigenmann C.H. 1908. Preliminary descriptions of new genera and species of Tetragonopterid characins. Bulletin of the Museum of Comparative Zoology 52: 91–106.
Eigenmann C.H. 1918. The American Characidae (Part 2). Memoirs of the Museum of Comparative Zoology 43: 101–208.
Fink W. & Weitzman S. 1974. The so called cheirodontin fishes of Central America with descriptions of two new species (Pisces: Characidae). Smithsonian Contributions to Zoology 172: 1–45. https://doi.org/10.5479/si.00810282.172
Fricke R. & Eschmeyer W.N. 2020. Guide to fish collections. Available from http://researcharchive.calacademy.org/research/ichthyology/catalog/collections.asp [accessed 10 Mar. 2020].
Fricke R., Eschmeyer W.N. & van der Laan R. 2020a. Catalog of Fishes: Genera, Species, References. Available from http://researcharchive.calacademy.org [accessed 31 Mar. 2020].
Fricke R., Eschmeyer W.N. & Jon D.F. 2020b. Catalog of Fishes: Species by Family/Subfamily. Available from http://researcharchive.calacademy.org/research/ichthyology/catalog/SpeciesByFamily.asp [accessed 31 Mar. 2020].
Fujisawa T. & Barraclough T.G. 2013. Delimiting species using single-locus data and the Generalized Mixed Yule Coalescent approach: a revised method and evaluation on simulated datasets. Systematic Biology 62 (5): 707–724. https://doi.org/10.1093/sysbio/syt033
García-Alzate C.A., Román-Valencia C. & Taphorn D.C. 2008. Revision of the Hyphessobrycon heterorhabdus-group (Teleostei: Characiformes: Characidae), with description of two new species from Venezuela. Vertebrate Zoology 58 (2): 139–157.
García-Melo J.E., Oliveira C., Da Costa Silva G.J., Ochoa-Orrego L.E., Garcia Pereira L.H. & Maldonado-Ocampo J.A. 2019. Species delimitation of neotropical Characins (Stevardiinae): Implications for taxonomy of complex groups. PLoS ONE 14 (6): e0216786. https://doi.org/10.1371/journal.pone.0216786
Géry J. 1977. Characoids of the World. TFH-publications, Neptune City Inc.
Goldstein P.Z. & Desalle R. 2010. Integrating DNA barcode data and taxonomic practice: determination, discovery, and description. BioEssays 33 (2): 135–147. https://doi.org/10.1002/bies.201000036
Guimarães E.C., De Brito P.S., Feitosa L.M., Carvalho-Costa L.F. & Ottoni F.P. 2018. A new species of Hyphessobrycon Durbin from northeastern Brazil: evidence from morphological data and DNA barcoding (Characiformes, Characidae). ZooKeys 765: 79–101. https://doi.org/10.3897/zookeys.765.23157
Guimarães E.C., De Brito P.S., Feitosa L.M., Carvalho-Costa L.F. & Ottoni F P. 2019. A new cryptic species of Hyphessobrycon Durbin, 1908 (Characiformes, Characidae) from the Eastern Amazon, revealed by integrative taxonomy. Zoosystematics and Evolution 95 (2): 345-360. https://doi.org/10.3897/zse.95.34069
Hebert P.D.N., Cywinska A., Ball S.L. & de Waard J.R. 2003a. Biological identifications through DNA barcodes. Proceedings of the Royal Society B 270 (1512): 313–321. https://doi.org/10.1098/rspb.2002.2218
Hebert P.D.N., Ratnasingham S. & de Waard J.R. 2003b. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society B 270 (1): 96–99. https://doi.org/10.1098/rsbl.2003.0025
Hebert P.D.N., Penton E.H., Burns J.M., Janzen D.H. & Hallwachs W. 2004a. Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences 101 (41): 14812–14817. https://doi.org/10.1073/pnas.0406166101
Hebert P.D.N., Stoeckle M.Y., Zemlak T.S. & Francis C.M. 2004b. Identification of birds through DNA barcodes. PLoS Biology 2 (10): e312. https://doi.org/10.1371/journal.pbio.0020312
Hrbek T., Da Silva V.M.F., Dutra N., Gravena W., Martin A.R. & Farias I.P. 2014. A new species of river dolphin from Brazil or: How little do we know our biodiversity. PLoS ONE 9 (1): e83623. https://doi.org/10.1371/journal.pone.0083623
Huelsenbeck J.P. & Ronquist F. 2001. Mr. Bayes: Bayesian inference of phylogenetic trees. Bioinformatics 17 (8): 754–755. https://doi.org/10.1093/bioinformatics/17.8.754
Jacobina U.P., Lima S.M.Q., Maia D.G., Souza G., Batalha-Filho H. & Torres R.A. 2018. DNA barcode sheds light on systematics and evolution of neotropical freshwater trahiras. Genetica 146 (6): 505–515. https://doi.org/10.1007/s10709-018-0043-x
Javonillo R., Malabarba L.R., Weitzman S.H. & J.R. Burns. 2010. Relationships among major lineages of characid fishes (Teleostei: Ostariophysi: Characiformes), based on molecular sequence data. Molecular Phylogenetics and Evolution 54 (2): 498–511. https://doi.org/10.1016/j.ympev.2009.08.026
Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution 16 (2): 111–120. https://doi.org/10.1007/BF01731581
Kumar S., Stecher G. & Tamura K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870–1874. https://doi.org/10.1093/molbev/msw054
Leary S., Underwood W., Anthony R., Cartner S., Corey D. & Grandin T. 2013. AVMA Guidelines for the Euthanasia of Animals. Available from: http://works.bepress. com/cheryl_greenacre/14 [accessed 31 Mar. 2020].
Malabarba L. R. & Weitzman S. H. 2003. Description of a new genus with six new species from Southern Brazil, Uruguay and Argentina, with a discussion of a putative characid clade (Teleostei: Characiformes: Characidae). Comunicações do Museu de Ciências e Tecnologia, PUCRS, Série Zoologia 16 (1): 67–151.
Mattos J.L.O. & Costa W.J.E.M. 2018. Three new species of the ‘Geophagus’ brasiliensis species group from the northeast Brazil (Cichlidae, Geophagini). Zoosystematics and Evolution 94 (2): 325–337. https://doi.org/10.3897/zse.94.22685
Melo B.F., Sidlauskas B.L., Hoekzema K., Vari R.P. & Oliveira C. 2014. The first molecular phylogeny of Chilodontidae (Teleostei: Ostariophysi: Characiformes) reveals cryptic biodiversity and taxonomic uncertainty. Molecular Phylogenetics and Evolution 70: 286–295. https://doi.org/10.1016/j.ympev.2013.09.025
Melo B.F., Ochoa L.E., Vari R.P. & Oliveira C. 2016a. Cryptic species in the Neotropical fish genus Curimatopsis (Teleostei, Characiformes). Zoologica Scripta 45: 650–658. https://doi.org/10.1111/zsc.12178
Melo B.F., Sidlauskas B.L., Hoekzema K., Frable B.W., Vari R.P. & Oliveira C. 2016b. Molecular phylogenetics of the Neotropical fish family Prochilodontidae (Teleostei: Characiformes). Molecular Phylogenetics and Evolution 102: 189–201. https://doi.org/10.1016/j.ympev.2016.05.037
Melo B.F., Benine R.C., Silva G.S.C., Avelino G.S. & Oliveira C. 2016c. Molecular phylogeny of the Neotropical fish genus Tetragonopterus (Teleostei: Characiformes: Characidae). Molecular Phylogenetics and Evolution 94: 709–717. https://doi.org/10.1016/j.ympev.2015.10.022
Mirande J.M. 2010. Phylogeny of the family Characidae (Teleostei: Characiformes): from characters to taxonomy. Neotropical Ichthyology 8 (3): 385–568. https://doi.org/10.1590/S1679-62252010000300001
Mirande J.M. 2018. Morphology, molecules and the phylogeny of Characidae (Teleostei, Characiformes). Cladistics 1–19. https://doi.org/10.1111/cla.12345
Murphy J.C., Jowers M.J., Lehtinen R.M., Charles S.P., Colli G.R., Peres Jr.A.K., Hendry C.R. & Pyron R.A. 2016. Cryptic, sympatric diversity in tegu lizards of the Tupinambis teguixin group (Squamata, Sauria, Teiidae) and the description of three new species. PLoS ONE 11 (8): e0158542. https://doi.org/10.1371/journal.pone.0158542
Oliveira C., Avelino G.S., Abe K.T., Mariguela T.C., Benine R.C., Ortí G., Vari R.P. & Castro R.M.C. 2011. Phylogenetic relationships within the speciose family Characidae (Teleostei: Ostariophysi: Characiformes) based on multilocus analysis and extensive ingroup sampling. BMC Evolutionary Biology, 11, 275. https://doi.org/10.1186/1471-2148-11-275
Ota R.R., Carvalho F.R. & Pavanelli C.S. 2020. Taxonomic review of the Hyphessobrycon panamensis species-group (Characiformes: Characidae). Zootaxa 4751 (3): 401–436. https://doi.org/10.11646/zootaxa.4751.3.1
Ottoni F.P., Mattos J.L.O., Katz A.M. & Bragança P.H.N. 2019. Phylogeny and species delimitation based on molecular approaches on the species of the Australoheros autrani group (Teleostei, Cichlidae), with biogeographic comments. Zoosystematics and Evolution 95 (1): 49–64. https://doi.org/10.3897/zse.95.31658
Padial J.M., Miralles A., De la Riva I. & Vences M. 2010. The integrative future of taxonomy. Frontiers in Zoology 7: 16. https://doi.org/10.1186/1742-9994-7-16
Pereira L.H.G., Pazian M.F., Hanner R., Foresti F. & Oliveira C. 2011. DNA barcoding reveals hidden diversity in the Neotropical freshwater fish Piabina argentea (Characiformes: Characidae) from the Upper Paraná basin of Brazil. Mitochondrial DNA 22 (1): 87–96. https://doi.org/10.3109/19401736.2011.588213
Poulakakis N., Edwards D.L., Chiari Y., Garrick R.C., Russello M.A., Benavides E., Watkins-Colwell G.J., Glaberman S., Tapia W., Gibbs J.P., Cayot L.J. & Caccone A. 2015. Description of a new Galapagos giant tortoise species (Chelonoidis; Testudines:Testudinidae) from Cerro Fatal on Santa Cruz Island. PLoS ONE 10 (10): e013877. https://doi.org/10.1371/journal.pone.0138779
Puillandre N., Lambert A., Brouillet S. & Achaz G. 2012. ABGD, automatic barcode gap discovery for primary species delimitation. Molecular Ecology 21: 1864–1877. https://doi.org/10.1111/j.1365-294X.2011.05239.x
Queiroz de K. 2005. Different species problems and their resolution. BioEssays 27 (12): 1263–1269. https://doi.org/10.1002/bies.20325
Queiroz de K. 2007. Species concepts and species delimitation. Systematic Biology 56 (6): 879–886. https://doi.org/10.1080/10635150701701083
Roca A.L., Georgiadis N.J., Pecon-Slattery J. & O’Brien S.J. 2001. Genetic evidence for two species of elephant in Africa. Science 80 (293): 1473–1477. https://doi.org/10.1126/science.1059936
Rosso J.J., González-Castro M., Bogan S., Cardoso Y.P., Mabragaña E., Delpiani M. & Astarloa J.M.D. 2018. Integrative taxonomy reveals a new species of the Hoplias malabaricus species complex (Teleostei: Erythrinidae). Ichthyological Exploration of Freshwaters IEF-1076: 1–18. https://doi.org/10.23788/IEF-1076
Rozas J., Sánchez J.C., Messeguer X. & Rozas R. 2003. DnaSP, DNA polymorphism analyses by the 506 coalescent and other methods. Bioinformatics 19: 2496–2497. https://doi.org/10.1093/bioinformatics/btg359
de Santana C.D., Crampton W.G., Dillman C.B., Frederico R.G., Sabaj M.H., Covain R., Ready J., Zuanon J., Oliveira R.R., Mendes Júnior R.N.G., Bastos D.A., Teixeira T.F., Mol J., Ohara W., Castro N.C., Peixoto L.A., Nagamachi C., Sousa L., Montag L.F.A., Ribeiro F., Waddell J.C., Piorsky N.M., Vari R.P. & Wosiacki W.B. 2019 Unexpected species diversity in electric eels with a description of the strongest living bioelectricity generator. Nature Communications 10: 4000. https://doi.org/10.1038/s41467-019-11690-z
Sites J.W. & Marshall J.C. 2003. Delimiting species: A Renaissance issue in systematic biology. Trends in Ecology and Evolution 18: 462–470. https://doi.org/10.1016/S0169-5347(03)00184-8
Swofford D.L. 2002. PAUP. Phylogenetic Analysis Using Parsimony (*and Other Methods), 4.0 edn, Sinauer, Sunderland, MA.
Sytsma K.J. & Schaal B.A. 1985. Phylogenetics of the Lisianthius skinneri (Gentianaceae) complex in Panama utilizing DNA restriction fragment analysis. Evolution 39: 594–608.
Taylor W.R. & Van Dyke G.C. 1985. Revised procedures for staining and clearing small fishes and other vertebrates for bone and cartilage study. Cybium 9 (2): 107–120.
Ward R.D., Zemlak T.S., Innes B.H., Las, P.R. & Hebert P.D.N. 2005. DNA barcoding Australia’s fish species. Philosophical Transactions of the Royal Society of London B, Biological Sciences 360 (1462): 1847–1857. https://doi.org/10.1098/rstb.2005.1716
Weitzman S.H. 1962. The osteology of Brycon meeki, a generalized characid fish, with an osteological definition of the family. Stanford Ichthyological Bulletin 8 (1): 3–77.
Weitzman S.H. & Palmer L. 1997. A new species of Hyphessobrycon (Teleostei: Characidae) from the Neblina region of Venezuela and Brazil, with comments on the putative ‘rosy tetra clade’. Ichthyological Exploration of Freshwaters 7 (3): 209–242.
Wiens J.J. & Penkrot T.A. 2002. Delimiting species using DNA and Morphological variation and discordant limits in spiny lizards (Sceloporus). Systematic biology 51 (1): 69–91. https://doi.org/10.1080/106351502753475880
Xia X.H. 2013. Dambe5: a comprehensive software package for data analysis in molecular biology and evolution. Molecular Biology and Evolution 30: 1720–1728. https://doi.org/10.1093/molbev/mst064
Xia X.H., Xie Z., Salemi M., Chen L. & Wang Y. 2003. An index of substitution saturation and its application. Molecular Phylogenetics and Evolution 26: 1–7. https://doi.org/10.1016/S1055-7903(02)00326-3
Zhang J., Kapli P., Pavlidis P. & Stamatakis A. 2013. A general species delimitation method with applications to phylogenetic placements. Bioinformatics 29: 2869–2876. https://doi.org/10.1093/bioinformatics/btt499
Copyright (c) 2020 Erick Cristofore Guimarães, Pâmella Silva de Brito, Pedro Henrique Negreiros Bragança, Jadson Pinheiro Santos, Axel Makay Katz, Luis Fernando Carvalho Costa, Felipe Polivanov Ottoni
This work is licensed under a Creative Commons Attribution 4.0 International License.
Creative Commons Copyright Notices
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (CC BY 4.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are NOT ALLOWED TO post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to taxonomic issues.