Skip to main content Skip to main navigation menu Skip to site footer
Published: 2007-05-28

Molecular systematics of Serrasalmidae: Deciphering the identities of piranha species and unraveling their evolutionary histories

Dept. of Biological Sciences, Florida International University, University Park, Miami, FL 33199, USA
United States Geological Survey, 7920 NW 71st St., Gainesville, FL 32653, USA
Dept. of Biological Sciences, Florida International University, University Park, Miami, FL 33199, USA
United States Geological Survey, 7920 NW 71st St., Gainesville, FL 32653, USA
Dept. of Biological Sciences, Florida International University, University Park, Miami, FL 33199, USA
Molecular Serrasalmidae piranhas Characidae Characiformes Serrasalmus Pristobrycon phylogeny polytomy biogeography


Piranhas and their relatives have proven to be a challenging group from a systematic perspective, with difficulties in identification of species, linking of juveniles to adults, diagnosis of genera, and recognition of higher-level clades. In this study we add new molecular data consisting of three mitochondrial regions for museum vouchered and photo-documented representatives of the Serrasalmidae. These are combined with existing serrasalmid sequences in GenBank to address species and higher-level questions within the piranhas using parsimony and Bayesian methods. We found robust support for the monophyly of Serrasalmus manueli, but not for Serrasalmus gouldingi when GenBank specimens identified as S. gouldingi were included in the analysis. “Serrasalmus gouldingi” sequences in GenBank may, however, be misidentified. Linking of juveniles to adults of the same species was greatly facilitated by the addition of sequence data. Based on our sampling and identifications, our data robustly reject the monophyly of the genera Serrasalmus and Pristobrycon. We found evidence for a well-supported clade comprised of Serrasalmus, Pygocentrus, and Pristobrycon (in part). This clade was robustly supported in separate and combined analyses of gene regions, and was also supported by a unique molecular character, the loss of a tandem repeat in the control region. Analysis of specimens and a literature review suggest this clade is also characterized by the presence of a pre-anal spine and ectopterygoid teeth. A persistent polytomy at the base of this clade was dated using an independent calibration as 1.8 million years old, corresponding to the beginning of the Pleistocene Epoch, and suggesting an origin for this clade more recent than dates cited in the recent literature. The sister group to this clade is also robustly supported, and consists of Catoprion, Pygopristis, and Pristobrycon striolatus. If the term piranha is to refer to a monophyletic clade, it should be restricted to Serrasalmus, Pygocentrus, and Pristobrycon (in part), or expanded to include these taxa plus Pygopristis, Catoprion, and Pristobrycon striolatus.


  1. Agostinho, C.S., Agostinho, A.A., Marques, E.E. & Bini, L.M. (1997) Abiotic factors influencing piranha attacks on netted fish in the upper Parana River, Brazil. North American Journal of Fisheries Management, 17, 712–718.

    Araujo-Lima, C. & Goulding, M. (1997) So Fruitful a Fish: Ecology, Conservation, and Aquaculture of the Amazon’s Tambaqui. Columbia University Press, New York, 191 pp.

    Arratia, G. & Cione, L.A. (1996) The fossil record of fossil fishes of Southern South America. In: Arratia, G. (Ed.), Contributions of Southern South America to Vertebrate Paleontology. Special volume of the Münchner Geowissenschaftliche Abhandlungen 30(A), pp. 9–72.

    Benson, G. (1999) Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Research, 27, 573–580.

    Braga, R.A. (1975) Ecologia e Etologia de Piranhas no Nordeste do Brasil (Pisces–Serrasalmus Lacépède, 1803). Banco do Nordeste do Brasil, Fortaleza, Ceará, Brazil, 268 pp.

    Braun, E.L. & Kimball, R.T. (2001) Polytomies, the power of phylogenetic inference, and the stochastic nature of molecular evolution: a comment on Walsh et al. (1999). Evolution, 55, 1261–1263.

    Calcagnotto, D., Schaefer, S.A. & DeSalle, R. (2005) Relationships among characiform fishes inferred from analysis of nuclear and mitochondrial gene sequences. Molecular Phylogenetics and Evolution, 36, 135–153.

    Castagnoli, N. (2000) Brazil fish culture. In: Stickney, R.R. (Ed.), Encyclopedia of Aquaculture. John Wiley and Sons, New York, pp. 123–128.

    Cestari, M.M. & Galetti Jr., P.M. (1992) Chromosome evolution in the genus Serrasalmus and cytotaxonomic considerations about Serrasalminae (Characidae, Pisces). Revista Brasileira de Genética, 15, 555–567.

    Donaldson, K.A. & Wilson, R. R. (1999) Amphi-panamic geminates of snook (Percoidei: Centropomidae) provide a calibration of the divergence rate in the mitochondrial DNA control region of fishes. Molecular Phylogenetics and Evolution, 13, 208–213.

    Eigenmann, C.H. (1915) The Serrasalminae and Mylinae. Annals of the Carnegie Museum, 9(3–4), 226–272.

    Fernández-Yepez, A. & Ramírez, M.V. (1967) Los caribes (serrasalmidos) de Venezuela y las pesqueras. Trabajos anexos a la Comisión Contribuciones al Tema, Ier Foro Internacional sobre Planificación y Desarrollo Pesquero, Caracas, Venezuela, 25 pp., figures. (mimeo).

    Fink, W.L. (1988) Evolutionary biology of piranhas. In: Dalrymple, G.H., Loftus, W.F. & Bernardino, F.S. (Eds.), Wildlife in the Everglades and Latin American Wetlands. Abstracts on the Proceedings of the First Everglades National Park Symposium, held February 25–March 1, 1985. Florida International University, Miami, Florida, pp. 13–14.

    Fink, W.L. (1989) Ontogeny and phylogeny of shape change and diet in the South American fishes called piranhas. In: David, B., Dommergues, J.-L., Chaline, J. & Laurin, B. (Eds.), Ontogenese et Évolution. Geobios, mémoire special, No. 12, 167–172.

    Fink, W.L. (1993) Revision of the piranha genus Pygocentrus (Teleostei, Characiformes). Copeia, 1993, 665–687.

    Fink, W.L. & Machado-Allison, A. (1992) Three new species of piranhas from Brazil and Venezuela (Teleostei: Characiformes). Ichthyological Exploration Freshwaters, 3, 55–71.

    Fink, W.L. & Zelditch, M.L. (1995) Phylogenetic analysis of ontogenetic shape transformations: a reassessment of the piranha genus Pygocentrus (Teleostei). Systematic Biology, 44, 343–360.

    Fink, W.L. & Zelditch, M.L. (1997) Shape analysis and taxonomic status of the Pygocentrus piranhas (Ostariophysi, Characiformes) from the Paraguay and Paraná river basins of South America. Copeia, 1997, 179–182.

    Frailey, C.D., Lavina, E.L., Rancy, A. & de Souza–Filho, J.P. (1988) A proposed Pleistocene/Holocene lake in the Amazon basin and its significance to Amazonian geology and biogeography. Acta Amazonica, 18, 119–143.

    Gaviria, J.I., Nirchio, M., Granado, A. & Estrada, A. (2005). Karyotype and nucleolar organizer regions of Pygocentrus cariba (Serrasalminae) from Caricara del Orinoco, Venezuela. Interciencia, 30, 44–47.

    Géry, J. (1972) Poissons characoides des Guyanes II. Famille des Serrasalmidae. Zoologische Verhandelingen Leiden, 122, 134–248.

    Géry, J. (1976) Les genres de Serrasalminae (Pisces–Characoidei). Bulletin Zoologisch Museum Universiteit van Amsterdam, 5, 47–54.

    Géry, J. (1977) Characoids of the World. Tropical Fish Hobbyist Publications, Neptune City, New Jersey, 672 pp.

    Goldman, N. & Whelan, S. (2000) Statistical tests of gamma-distributed rate heterogeneity in models of sequence evolution in phylogenetics. Molecular Biology and Evolution, 17, 975–978.

    Gosline, W.A. (1951) Notes on the characid fishes of the subfamily Serrasalminae. Proceedings of the California Academy of Sciences, 27, 17–64.

    Goulding, M. (1980) The Fishes and the Forest: Explorations in Amazonian Natural History. University of California Press, Berkeley, 280 pp.

    Goulding, M., Carvalho, M.L. & Ferreira, E.G. (1988) Rio Negro, Rich Life in Poor Water. SPB Academic Publishing, The Hague, The Netherlands, 200 pp.

    Haddad, V., Jr. & Sazima, I. (2003) Piranha attacks on humans in southeast Brazil: epidemiology, natural history, and clinical treatment, with description of a bite outbreak. Wilderness and Environmental Medicine, 14, 249–254.

    Hubert, N., Duponchelle, F., Nuñez, J., Rivera, R. & Renno, J.-F. (2006) Evidence of reproductive isolation among closely related sympatric species of Serrasalmus (Ostariophysii, Characidae) from the Upper Madeira River, Amazon, Bolivia. Journal of Fish Biology, 69, 31–51.

    Jégu, M. (1992) Variations du niveau marin et distribution des poisons d’eau douce en Amazonie Orientale. In: Prost, M.T. (Ed.), Evolution des littoraux de Guyane et de la zone caraïbe méridionale pendant le quaternaire. ORSTOM, Paris, pp. 281–297.

    Jégu, M. (2003) Subfamily Serrasalminae (pacus and piranhas). In: Reis, R.E., Kullander, S.O. & Ferraris, C.J., Jr. (Eds.), Check list of Freshwater Fishes of South and Central America. Edipucrs, Porto Alegre, Brazil, pp. 182–196.

    Jégu, M. & Dos Santos, G.M. (1988) Le genre Serrasalmus (Pisces, Serrasalmidae) dans le bas Tocantins (Brésil, Parà), avec la description d’une espèce nouvelle, S. geryi, du basin Araguaia–Tocantins. Revue d'Hydrobiologie Tropicale, 21, 239–274.

    Jégu, M. & Dos Santos, G.M. (2001) Mise au point à propos de Serrasalmus spilopleura Kner, 1858 et réhabilitation de S. maculatus Kner, 1858. Cybium, 25, 119–143.

    Jégu, M., Leão, E.L.M. & Dos Santos, G.M. (1991) Serrasalmus compressus, une espece nouvelle du Rio Madeira, Amazonie (Pisces: Serrasalmidae). Ichthyological Exploration of Freshwaters, 2, 97–108.

    Kocher, T.D., Thomas, W.K., Meyer, A., Edwards, S.V., Paabo, S., Villablanca, F.X. & Wilson, A.C. (1989) Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proceedings of the National Academy of Science of the United States of America, 86, 6196–6200.

    Lasso, C.A., Mojica, J.I., Usma, J.S., Maldonado-O., J.A., DoNascimiento, C., Taphorn, D.C., Provenzano, F., Lasso-Alcala, O., Galvis, G., Vasquez, L., Lugo, M., Machado-Allison, A., Royero, R., Suárez, C. & Ortega-Lara, A. (2004). Peces de la cuenca del río Orinoco. Parte I: lista de especies y distribución por subcuencas. Biota Colombiana 5 (2), 95–158.

    Lovejoy, N.R. 1997. Stingrays, parasites, and neotropical biogeography: a closer look at Brooks et al.’s hypotheses concerning the origins of neotropical freshwater rays (Potamotrygonidae). Systematic Biology, 46, 218–230.

    Ludwig, A., May, B., Debus, L. & Jenneckens, I. (2000) Heteroplasmy in the mtDNA control region of sturgeon (Acipenser, Huso and Scaphirhynchus). Genetics, 156, 1933–1947.

    Lundberg, J.G., Machado-Allison, A. & Kay, R.F. (1986) Miocene characid fishes from Colombia: evolutionary stasis and extirpation. Science, 234, 208–209.

    Lundberg, J.G. (1997) Freshwater fishes and their paleobiotic implications. In: Kay, R.F., Madden. R.L., Cifelli, R.L. & Flynn, J.J (Eds.), Vertebrate Paleontology in the Neotropics: The Miocene Fauna of La Venta, Columbia. Smithsonian Institution Press, Washington, D.C., pp. 67–91.

    Lundberg, J.G. (1998) The temporal context for the diversification of neotropical fishes. In: Malabarba, L.R., Reis, R.E., Vari, R.P., Lucena, Z.M.S. & Lucena, C.A.S. (Eds.), Phylogeny and Classification of Neotropical Fishes. Edipucrs, Porto Alegre, Brazil, pp. 49–68.

    Lunt, D.H., Whipple, L.E. & Hyman, B.C. (1998) Mitochondrial DNA variable number tandem repeats (VNTRs): utility and problems in molecular ecology. Molecular Ecology, 7, 1441–1455.

    Machado-Allison, A. (1982) Studies on the systematics of the subfamily Serrasalminae (Pisces–Characidae). Unpublished Ph.D. dissertation, The George Washington University, Washington D.C.

    Machado-Allison, A. (1983) Estudios sobre la sistematica de la subfamilía Serrasalminae (Teleostei, Characidae). Parte 2. Discusión sobre la condición monofilética de la subfamilía. Acta Biologica Venezuelica, 11, 145–195.

    Machado-Allison, A. (1985) Estudios sobre la sistemática de la subfamilia Serrasalminae. Parte III. Sobre el estatus generico y relaciones filogenéticas de los géneros Pygopristis, Pygocentrus, Pristobrycon y Serrasalmus (Teleostei–Characidae–Serrasalminae). Acta Biologica Venezuelica, 12, 19–42.

    Machado-Allison, A. (2002) Los peces caribes de Venezuela: una aproximación a su estudio taxonómico. Boletín de la Academia de Ciencias Físicas, Matemáticas y Naturales de Venezuela, 62(1), 35–88.

    Machado-Allison, A. & Fink, W. (1995) Sinopsis de las especies de la subfamilia Serrasalminae presentes en la cuenca del Orinoco: claves, diagnosis e ilustraciones. Serie Peces de Venezuela, Universidad Central de Venezuela, Caracas, 90 pp.

    Machado-Allison, A. & Fink, W. (1996) Los Peces Caribes de Venezuela: Diagnosis, Claves, Aspectos Ecologicos y Evolutivos. Collection Monograph 52, Universidad Central de Venezuela, Caracas, 151 pp.

    Machado-Allison, A., Fink, W. & Antonio, M. E. (1989) Revisión del género Serrasalmus Lacepede, 1803 y géneros relacionados en Venezuela: I. Notas sobre la morfología y sistematica de Pristobrycon striolatus (Steindachner, 1908). Acta Biologica Venezuelica, 12, 140–171.

    Merckx, A., Jégu, M. & Dos Santos, G.M. (2000). Une nouvelle espèce de Serrasalmus (Teleostei: Characidae: Serrasalminae), S. altispinis n. sp., décrite du rio Uatumã (Amazonas, Brésil) avec une description complémentaire de S. rhombeus (Linnaeus, 1766) du Plateau Guyanais. Cybium, 24, 181–121.

    Nakayama, C.M., Jégu, M., Porto, J.I.R. & Feldberg, E. (2001) Karyological evidence for a cryptic species of piranha within Serrasalmus rhombeus (Characidae, Serrasalminae) in the Amazon. Copeia, 2001, 866–869.

    Nakayama, C.M., Porto, J.I.R. & Feldberg, E. (2002) A comparative cytogenetic study of five piranha species (Serrasalmus, Serrasalminae) from the Amazon basin. Genetica, 114, 231–236.

    Nico, L.G. (1991) Trophic ecology of piranhas (Characidae: Serrasalminae) from savanna and forest regions in the Orinoco River basin of Venezuela. Unpublished Ph.D. dissertation, University of Florida, Gainesville, 210 pp.

    Nico, L.G. & Taphorn, D.C. (1986) Those bitin’ fish from South America. Tropical Fish Hobbyist, 34 (Feb.), 24–27, 30–34, 36, 40–41, 56–57.

    Nico, L.G. & Taphorn, D.C. (1988) Food habits of piranhas in the low Llanos of Venezuela. Biotropica, 20, 311–321.

    Nirchio, M., Granado, A., Ron, E. & Pérez, J.E. (2005). Karyotype and nucleolar organizer regions in Serrasalmus rhombeus (Serrasalminae) from Caricara del Orinoco, Venezuela. Interciencia, 27, 676–678.

    Norman, J.R. (1929) The South American characid fishes of the subfamily Serrasalmoninae, with a revision of the genus Serrasalmus Lacepède. Proceedings of the Zoological Society of London, 52 (1928), 781–829.

    Oliveira C., Almeida-Toledo L.F., Foresti F., Britski H. & Toledo-Filho S.A. (1988) Chromosome formulae of Neotropical freshwater fishes. Revista Brasileira de Genética, 11, 577–624.

    Ortí, G., Petry, P., Porto, J.I.R., Jégu, M. & Meyer, A. (1996) Patterns of nucleotide change in mitochondrial ribosomal RNA genes and the phylogeny of piranhas. Journal of Molecular Evolution, 42, 169–182.

    Ortí, G., Porto, J.I.R. & Sivasundar, A. (2000) Phylogeny of the Serrasalminae (Characiformes) based on mitochondrial DNA sequences. Unpublished poster presented at the 80th Annual Meeting of the American Society of Ichthyologists and Herpetologists, La Paz, Baja California Sur, Mexico (14–20 June 2000). Available from (accessed 30 May 2005)

    Palumbi, S.R. (1996) Nucleic acids II: The polymerase chain reaction. In: Hillis, D.M., Moritz, C. & Mable, B.K. (Eds.), Molecular Systematics, 2nd edition. Sinauer Associates, Sunderland, Massachusetts, pp. 205–247.

    Porto, J.I.R., Feldberg, E., Nakayama, C.M., Maia, R.O. & Jégu, M. (1991) Cytotaxonomic analysis in the Serrasalmidae (Ostariophysi, Characiformes). Abstracts of presentations, Seventh International Ichthyology Congress of the European Ichthyological Union, The Hague, The Netherlands. Bulletin Zoologisch Museum, Universiteit van Amsterdam (special issue), 66.

    Porto, J.I.R., Feldberg E., Nakayama, C. & Falcao, J.N. (1992) A checklist of chromosome numbers and karyotypes of Amazonian freshwater fishes. Revue d'Hydrobiologie Tropicale, 25, 287–299.

    Posada D. & Crandall, K.A. 1998. Modeltest: testing the model of DNA substitution. Bioinformatics, 14, 817–818.

    Raffles, H. & Winkler-Prins, A.M.G.A. (2003) Further reflections on Amazonian environmental history: transformations of rivers and streams. Latin American Research Review, 38, 165–187.

    Ravago, R.G., Monje, V.D. & Juinio-Menez, M.A. (2002) Length and sequence variability in mitochondrial control region of the Milkfish, Chanos chanos. Marine Biotechnology, 4, 40–50.

    Ray, D.A. & Densmore, L. (2002) The crocodilian mitochondrial control region: General structure, conserved sequences, and evolutionary implications. Journal of Experimental Zoology, 294, 334–345.

    Reis, R.E. (1998) Systematics, biogeography, and the fossil record of the Callichthyidae: a review of the available data. In: Malabarba, L.R., Reis, R.E., Vari, R.P., Lucena, Z.M.S. & Lucena, C.A.S. (Eds.), Phylogeny and Classification of Neotropical Fishes. Edipucrs, Porto Alegre, Brazil, pp. 351–374.

    Ronquist, F. & Huelsenbeck, J. (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572–1574.

    Rubilar, A. (1994) Diversidad ictiológica en depósitos continentales miocenos de la Formación Cura–Mallín, Chile (37–39°S): implicancias paleogeográficas. Revista Geológica de Chile, 21, 3–29.

    Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A. & Allard, R.W. (1984) Ribosomal DNA spacer length in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Science of the United States of America, 81, 8014–8018.

    Sazima, I. & Andrade–Guimaraes, S. (1987) Scavenging on human corpses as a source for stories about man–eating piranhas. Environmental Biology of Fishes, 20, 75–77.

    Sazima, I. & Machado, F.A. (1990) Underwater observations of piranhas in western Brazil. Environmental Biology of Fishes, 28, 17–31.

    Sivasundar, A., Bermingham, E. & Ortí , G. (2001) Population structure of migratory freshwater fishes (Prochilodus: Characiformes) in major South American rivers. Molecular Ecology, 10, 407–418.

    Slowinski, J.B. (2001) Molecular polytomies. Molecular Phylogenetics and Evolution, 19, 114–120.

    Stern, K.M. (1970) Der Casiquiare–Kanal, einst und jetzt. Amazoniana, 2, 401–416.

    Sternberg, H.O. (1975) The Amazon River of Brazil. Geographische Zeitschrift, 40, 1–74.

    Swofford, D.L. (2002) PAUP*: Phylogenetic Analysis Using Parsimony (*and Other Methods). Sinauer Associates, Sunderland, Massachusetts.

    Thompson, J.D., Gibson T.J., Plewniak F., Jeanmougin F. & Higgins D.G. (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24), 4876–82.

    Van Every, L.R. & Kritsky, D.C. (1992) Neotropical Monogenoidea. 18 Anacanthorus Mizelle and Price, 1965 (Dactylogyridae, Anacanthorinae) of piranha (Characoidea, Serrasalmindae) from the Central Amazon, their phylogeny, and aspects of host–parasite coevolution. Journal of Helminthological Society of Washington, 59, 52–75.

    Walsh, H.E. & Friesen, V.L. (2001) Power and stochasticity in the resolution of soft polytomies: A reply to Braun et al. Evolution, 55, 1264–1266.

    Walsh, H.E., Kidd, M.G., Moum, T. & Friesen, V.L. (1999) Polytomies and the power of phylogenetic inference. Evolution, 53, 932–937.

    Winemiller, K.O. (1989) Ontogenetic diet shifts and resource partitioning among piscivorous fishes in the Venezuelan llanos. Environmental Biology of Fishes, 26, 177–199.