Abstract
Anurans are renowned for a high diversity of reproductive modes, but less than 1 % of species exhibit internal fertilisation followed by viviparity. In the live-bearing West African Nimba toad (Nimbaphrynoides occidentalis), females produce yolk-poor eggs and internally nourish their young after fertilisation. Birth of fully developed juveniles takes place after 9 months. In the present study, we used genetic markers (eight microsatellite loci) to assign the paternity of litters of 12 females comprising on average 9.7 juveniles. In 9 out of 12 families (75 %), a single sire was sufficient; in three families (25 %), more than one sire was necessary to explain the observed genotypes in each family. These findings are backed up with field observations of male resource defence (underground cavities in which mating takes place) as well as coercive mating attempts, suggesting that the observed moderate level of multiple paternity in a species without distinct sperm storage organs is governed by a balance of female mate choice and male reproductive strategies.
This is a preview of subscription content, access via your institution.

References
Adams EM, Jones AG, Arnold SJ (2005) Multiple paternity in a natural population of a salamander with long-term sperm storage. Mol Ecol 14:1803–1810
Angel F, Lamotte M (1947) Note sur la biologie d’un crapaud vivipare Nectophrynoides occidentalis Ang. C R Hebd Seances Acad Sci 224:413–415
Arnqvist G (1989) Multiple mating in a water strider: mutual benefits or intersexual conflict? Anim Behav 38:749–756
Arnqvist G, Kirkpatrick M (2005) The evolution of infidelity in socially monogamous passerines: the strength of direct and indirect selection on extrapair copulation behavior in females. Am Nat 165(Suppl):S26–S37
Ashby B, Gupta S (2013) Sexually transmitted infections in polygamous mating systems. Philos Trans R Soc B 368:20120048
Birkhead TR (1998) Cryptic female choice: criteria for establishing female sperm choice. Evolution 52:1212–1218
Blackburn DG (1999) Viviparity and oviparity: evolution and reproductive strategies. Encycl Reprod 4:994–1003
Bouwman KM, Burke T, Komdeur J (2006) How female reed buntings benefit from extra-pair mating behaviour: testing hypotheses through patterns of paternity in sequential broods. Mol Ecol 15:2589–2600
Brown JL, Morales V, Summers K (2010) A key ecological trait drove the evolution of biparental care and monogamy in an amphibian. Am Nat 175:436–446
Buckley D (2012) Evolution of viviparity in salamanders (Amphibia, Caudata). Encyclopedia of Life Sciences. Wiley, Chichester, pp 1–13
Byrne PG, Roberts JD (1999) Simultaneous mating with multiple males reduces fertilization success in the myobatrachid frog Crinia georgiana. Proc R Soc Lond Ser B Biol Sci 266:717–721
Byrne PG, Roberts JD (2012) Evolutionary causes and consequences of sequential polyandry in anuran amphibians. Biol Rev 87:209–228
Byrne PG, Whiting MJ (2011) Effects of simultaneous polyandry on offspring fitness in an African tree frog. Behav Ecol 22:385–391
Caspers BA, Krause ET, Hendrix R, Kopp M, Rupp O, Rosentreter K, Steinfartz S (2014) The more the better - polyandry and genetic similarity are positively linked to reproductive success in a natural population of terrestrial salamanders (Salmandra salamandra). Mol Ecol 23:239–250
Castanet J, Pinto S, Loth M-M, Lamotte M (2000) Âge individuel, longévité et dynamique de croissance osseuse chez un amphibien vivipare, Nectophrynoides occidentalis (Anuoure, Bufonidé). Ann Sci Nat Zool Biol Anim 24:11–17
Chen YH, Cheng WC, Yu HT, Kam YC (2011) Genetic relationship between offspring and guardian adults of a rhacophorid frog and its care effort in response to paternal share. Behav Ecol Sociobiol 65:2329–2339
Duellman WE, Trueb L (1986) Biology of amphibians. The Johns Hopkins University Press, Baltimore
Dziminski MA, Roberts JD, Beveridge M, Simmons LW (2010) Among-population covariation between sperm competition and ejaculate expenditure in frogs. Behav Ecol 21:322–328
Frost DR (2015) Amphibian species of the world: an online reference. Version 6.0. http://research.amnh.org/herpetology/amphibia/index.html
Gavaud J (1976) La gemétogenèse du mâle de Nectophrynoides occidentalis Angel (Amphibien Anoure vivipare). I. - Étude quantitative au cours du cycle annuel chez l’adulte. Ann Biol Anim Biochim Biophys 16:1–12
Grafe TU, Stewart MM, Lampert KP, Rödel M-O (2011) Putting toe clipping into perspective: a viable method for marking anurans. J Herpetol 45:28–35
Gray EM (1997) Female red-winged blackbirds accrue material benefits from copulating with extra-pair males. Anim Behav 53:625–639
Haddad CFB, Prado CPA (2005) Reproductive modes in frogs and their unexpected diversity in the Atlantic Forest of Brazil. Bioscience 55:207–217
Hase K, Shimada M (2014) Female polyandry and size-assortative mating in isolated local populations of the Japanese common toad Bufo japonicus. Biol J Linn Soc 113:236–242
Hillers A, Loua NS, Rödel M-O (2008) Assessment of the distribution and conservation status of the viviparous toad Nimbaphrynoides occidentalis on Monts Nimba. Guinea. Endanger Species Res 5:13–19
Hudson CM, Fu J (2013) Male-biased sexual size dimorphism, resource defense polygyny, and multiple paternity in the Emei moustache toad (Leptobrachium boringii). PLoS ONE 8:e67502
Iskandar DT, Evans BJ, Mcguire JA (2014) A novel reproductive mode in frogs: a new species of fanged frog with internal fertilization and birth of tadpoles. PLoS ONE 9, e115884
Jehle R, Sztatecsny M, Wolf JBW, Whitlock A, Hödl W, Burke T (2007) Genetic dissimilarity predicts paternity in the smooth newt (Lissotriton vulgaris). Biol Lett 3:526–528
Johnson SL, Brockmann HJ (2013) Parental effects on early development: testing for indirect benefits of polyandry. Behav Ecol 24:1218–1228
Jones AG (2005) GERUD 2.0: a computer program for the reconstruction of parental genotypes from half-sib progeny arrays with known or unknown parents. Mol Ecol Notes 5:708–711
Jones OR, Wang J (2010) COLONY: a program for parentage and sibship inference from multilocus genotype data. Mol Ecol Resour 10:551–555
Knopp T, Merilä J (2009) Multiple paternity in the moor frog, Rana arvalis. Amphibia-Reptilia 30:515–521
Kupfer A, Wilkinson M, Gower DJ, Müller H, Jehle R (2008) Care and parentage in a skin-feeding caecilian amphibian. J Exp Zool 309A:460–467
Kusano T, Toda M, Fukuyama K (1991) Testes size and breeding systems in Japanese anurans with special reference to large testes in the treefrog, Rhacophorus arboreus (Amphibia: Rhacophoridae). Behav Ecol Sociobiol 29:27–31
Lamotte M (1959) Observations écologiques sur les populations naturelles de Nectophrynoides occidentalis (Fam. Bufonidés). Bull Biol Fr Belg 4:355–413
Lamotte M, Xavier F (1972) Les amphibiens anoures a développement direct d’Afrique. Observations sur la biologie de Nectophrynoides tornieri (Roux). Bull la Société Zool Fr 97:413–428
Lamotte M, Rey P, Vogeli M (1964) Recherches sur l’ovaire de Nectophrynoides occidentalis, batracien anoure vivipare. Arch Anat Microsc Morphol Exp 53:179–224
Larivière S, Ferguson SH (2003) Evolution of induced ovulation in North American carnivores. J Mammal 84:937–947
Lodé T, Lesbarrères D (2004) Multiple paternity in Rana dalmatina, a monogamous territorial breeding anuran. Naturwissenschaften 91:44–47
Lodé T, Holveck MJ, Lesbarrères D (2005) Asynchronous arrival pattern, operational sex ratio and occurrence of multiple paternities in a territorial breeding anuran, Rana dalmatina. Biol J Linn Soc 86:191–200
Nichols HJ, Cant MA, Sanderson JL (2015) Adjustment of costly extra-group paternity according to inbreeding risk in a cooperative mammal. Behav Ecol 26:1486–1494
Orr TJ, Brennan PLR (2015) Sperm storage: distinguishing selective processes and evaluating criteria. Trends Ecol Evol 30:261–272
Parker GA, Birkhead TR (2013) Polyandry: the history of a revolution. Philos Trans R Soc B 368:20120335
Pizzari T, Wedell N (2013) The polyandry revolution. Philos Trans R Soc B 368:20120041
Plough LV, Moran A, Marko P (2014) Density drives polyandry and relatedness influences paternal success in the Pacific gooseneck barnacle, Pollicipes elegans. BMC Evol Biol 14:81
Rovelli V, Randi E, Davoli F, Macale D, Bologna MA, Vignoli L, Tre R, Marconi V, Scienze D (2015) She gets many and she chooses the best: polygynandry in Salamandrina perspicillata (Amphibia: Salamandridae). Biol J Linn Soc 116:671–683
Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics Methods and Protocols: Methods in Molecular Biology. Humana Press, Totowa, pp 365–386
Sandberger-Loua L, Doumbia J, Rödel M-O (2016) Conserving the unique to save the diverse—identifying key environmental determinants for the persistance of the viviparous Nimba toad in a West African World Heritage Site. Biol Conserv 198:15–21
Shine R (1987) The evolution of viviparity: ecological correlates of reproductive mode within a genus of Australian snakes (Pseudechis: Elapidae). Copeia 1987:551–563
Simmons LW (2005) The evolution of polyandry: sperm competition, sperm selection and offspring viability. Annu Rev Ecol Evol Syst 36:125–146
Starr C (1984) Sperm competition, kinship, and sociality in aculeate Hymenoptera. In: Smith R (ed) Sperm competition and the evolution of animal mating systems. Academic Press, New York, pp 428–459
Sztatecsny M, Jehle R, Burke T, Hödl W (2006) Female polyandry under male harassment: the case of the common toad (Bufo bufo). J Zool 270:517–522
Uller T, Olsson M (2008) Multiple paternity in reptiles: patterns and processes. Mol Ecol 17:2566–2580
Vieites DR, Nieto-Román S, Barluenga M, Palanca A, Vences M, Meyer A (2004) Post-mating clutch piracy in an amphibian. Nature 431:305–308
Vilter V, Lugand A (1959) Trophisme intra-utérin et croissance embryonnaire chez le Nectophrynoides occidentalis Ang., crapaud totalement vivipare du Mont Nimba (Haute-Guinée). C R Seances Soc Biol Fil 153:29–32
Wake MH (1978) The reproductive biology of Eleutherodactylus jasperi (Amphibia, Anura, Leptodactylidae), with comments on the evolution of live-bearing systems. J Herpetol 12:121–133
Wake MH (1993) Evolution of oviductal gestation in amphibians. J Exp Zool 266:394–413
Wake MH (2015a) Fetal adaptations for viviparity in amphibians. J Morphol 276:941–960
Wake MH (2015b) How do homoplasies arise? Origin and maintenance of reproductive modes in amphibians. In: Dial KP, Shubin N, Brainerd EL (eds) Great transformations in vertebrate evolution. The University of Chicago Press, Chicago, pp 375–394
Wells KD (2010) The ecology and behavior of amphibians. The University of Chicago Press
Wood SN (2011) Fast stable restricted maximum likelihood and marginal likelihood estimation of semiparametric generalized linear models. J R Stat Soc Ser B 73:3–36
Wourms JP, Lombardi J (1992) Reflections on the evolution of piscine viviparity. Integr Comp Biol 32:276–293
Xavier F (1971) Recherches sur l’endocrinologie sexuelle de la femelle de Nectophrynoides occidentalis Angel (amphibien anoure vivipare). Faculté des sciences Paris, Thèse de doctorat d’état ès-Sciences Naturelles
Xavier F (1974) La pseudogestation chez Nectophrynoides occidentalis Angel. Gen Comp Endocrinol 22:98–115
Xavier F (1977) An exceptional reproductive strategy in anura: Nectophrynoides occidentalis Angel (Bufonidae), an example of adaptation to terrestrial life by viviparity. In: Hecht MK, Goody PC, Hecht BM (eds) Major patterns in vertebrate evolution. Vol 14. NATO advanced Study Institude, Series A, Life Sciences, New York pp 545–552
Xavier F (1986) La reproduction des Nectophrynoides. In: Grassé P-P, Delsol M (eds) Traité de Zoologie - anatomie, sysématique, biologie - Batraciens. Masson, Paris, pp 497–513
Xavier F, Zuber-Vogeli M, Le Quang Trong Y (1970) Recherches sur l’activité endocrine de l’ovaire de Nectophrynoides occidentalis Angel (Amphibien Anoure vivipare) - I. Etude histochimique. Gen Comp Endocrinol 15:425–431
Zhao M, Li C, Zhang W, Wang H, Luo Z, Gu Q, Gu Z, Liao C, Wu H (2016) Male pursuit of higher reproductive success drives female polyandry in the Omei treefrog. Anim Behav 111:101–110
Acknowledgments
We thank the Société de Mines de Fer, Guinée (SMFG) for financial and logistic support. For support in the field, we thank M. Hirschfeld, J. Doumbia, K. Camara, F. Gbêmou, B. Pivi and B. Doré. For helpful discussions, we thank Linus Günther and Simon Ripperger. We thank three anonymous reviewers for their constructive criticism, improving a previous draft of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
The study was funded by the Société des Mines de Fer de Guinée (SMFG), but the company had no influence on study design, data collection, data analyses and interpretation, writing of the manuscript and in the decision to submit the paper for publication; thus, the authors declare that they have no conflict of interests.
All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All work complies with the guidelines for the use of live amphibians and reptiles in research compiled by the American Society of Ichthyologists and Herpetologists (ASIH), The Herpetologists’ League (HL) and the Society for the Study of Amphibians and Reptiles (SSAR), as well as to the IUCN policy statement on research involving species at risk of extinction. The Ministère de l’Enseignement Superieur et de la Recherche Scientifique (MESRS) and the Direction Nationale de la Recherche Scientifique et Technique (DNRST) granted research permits (No. 085/DNPN/2007, No. 103/DNRSIT/DN, No. 095/MENSRS/DNRST No. 091/MESRS/DNRST/2009; No. 121/MESRS/DNRST/2010; No. 177/MESRS/DNRST/2011, No. 027/MESRS/DNRST/2012, No. 061/DNRSIT/DN and No. 020/MESRS/DNRSIT/2014). The authorities from the Ministère de l’Environement et du Development durable, Conakry and the Bundesamt für Naturschutz, Bonn granted CITES export (No. 00314, N°00492) and import permits (E-3117; E-4074), respectively.
Additional information
Communicated by: Sven Thatje
Electronic supplementary material
Below is the link to the electronic supplementary material.
Online Resource 1
Video1.avi, defending cavity entrance: shows a behavioural observation of a male defending a cavity entrance against another male. (AVI 2089 kb)
Online Resource 2
Video2.avi, antagonistic behaviours: shows behavioural observations of male antagonistic behaviours as aggressive calling and fighting. (AVI 3664 kb)
Online Resource 3
Video3.avi: male harassment: shows a male harassing a female, as well as a male trying to dislodge an amplected male. (AVI 2243 kb)
Online Resource 4
Video4.avi: female dislodges male: gives an example of a female dislodging an amplected male from her back. (AVI 2985 kb)
Rights and permissions
About this article
Cite this article
Sandberger-Loua, L., Feldhaar, H., Jehle, R. et al. Multiple paternity in a viviparous toad with internal fertilisation. Sci Nat 103, 51 (2016). https://doi.org/10.1007/s00114-016-1377-9
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00114-016-1377-9
Keywords
- Multiple paternity
- Internal fertilisation
- Operational sex ratio
- Male harassment
- Amphibia
- Nimbaphrynoides occidentalis