Journal of Insect Behavior

, Volume 31, Issue 6, pp 616–628 | Cite as

Patterns of Sperm Transfer Behavior in a Pholcid Spider with Two Distinct Copulatory Phases

  • Franco Cargnelutti
  • Lucia Calbacho-Rosa
  • Alex Córdoba-Aguilar
  • Alfredo V. PerettiEmail author


Sexual selection is the responsible force for the evolution and maintenance of genital diversity and function. This is the case for example, of genital movements performed by males during mating and copulation duration. Spiders perform ritualized copulations whereby males carry out different types of movements using their pedipalps with varying duration. The function and duration of these pedipalp movements is unclear. In the pholcid spider, Holocnemus pluchei males that copulate with virgin females perform two copulatory phases: phase I in which the pedipalps move and phase II in which pedipalps remain motionless. Using H. pluchei as a study species, our study aims were: 1) to assess if sperm transfer occurs when pedipalps move or are still and quantify the number of sperm in male bulbs and in the female uterus externus after copulation; and, 2) to determine if amount of sperm transferred to females is associated with duration of each copulatory phase. Two experimental groups (i. e. complete copulation and interrupted copulation) were established in which the amount of sperm remaining in the male bulbs and the amount of sperm stored by females were determined. Our results show that sperm transfer occurs during phase I, that males transfer almost all sperm from their bulbs while the females store only 20% of that male amount. There was no relation between the amount of sperm transferred or stored and the duration of the copulatory phases. These results support the hypothesis that while both phases may serve a copulatory courtship, only phase I (when pedipalps move) serves for sperm transfer.


Mating sperm transfer spermatozoa Holocnemus pluchei sexual selection 



We thank Camilo I. Mattoni, Alejandra Ceballos and Matias Izquierdo for their useful comments to previous versions of this paper. We also thank Fedra Bollatti, Macarena González, David Vrech, Paola Olivero and Natalia Rivetti for their help during collecting trips. We also thank the Jardín Zoológico de Córdoba and the Universidad Nacional de Córdoba for allowing us to collect the samples. Mathias Foellmer and an anonymous referee provided helpful and constructive comments which improved the manuscript. Financial support was provided by the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Fondo para la Investigación Científica y Tecnológica (FONCYT) and Secretaría de Ciencia y Tecnología de la Universidad Nacional de Córdoba (SECYT).

Compliance with Ethical Standards

This study was conducted in compliance with the “Guidelines for the use of animals in research” as published in Animal Behaviour (1991, 41, 183–186), and with the laws of the country where the research was conducted.

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

10905_2018_9702_MOESM1_ESM.avi (12.1 mb)
Online Resource 1 Male carrying out Phase I. (AVI 12373 kb)
10905_2018_9702_MOESM2_ESM.avi (8.8 mb)
Online Resource 2 Male carrying out Phase II. (AVI 9019 kb)


  1. Aisenberg A, Costa FG (2005) Females mated without sperm transfer maintain high sexual receptivity in the wolf spider Schizocosa malitiosa. Ethology 111:545–558CrossRefGoogle Scholar
  2. Albo MJ, Costa FG (2017) Female wolf spiders exert cryptic control drastically reducing ejaculate size. Ethology 123:659–666CrossRefGoogle Scholar
  3. Albo MJ, Peretti AV (2015) Worthless and nutritive nuptial gifts: mating duration, sperm stored and potential female decisions in spiders. PLoS One 10:e0129453CrossRefGoogle Scholar
  4. Albo MJ, Bilde T, Uhl G (2013) Sperm storage mediated by cryptic female choice for nuptial gifts. Proc R Soc Biol Sci Ser B 280:20131735CrossRefGoogle Scholar
  5. Alcock J (1994) Post-insemination associations between male and female insects: the mate-guarding hypothesis. Annu Rev Entomol 39:1–21CrossRefGoogle Scholar
  6. Assis BA, Foellmer MW (2016) One size fits all? Determinants of sperm transfer in a highly dimorphic orb-web spider. J Evol Biol 29:1106–1120CrossRefGoogle Scholar
  7. Blumstein DT, Evans CS, Daniel JC (2000) JWatcher.
  8. Bukowski TC, Christenson TR (1997) Determinants of sperm release and storage in a spiny orbweaving spider. Anim Behav 53:381–395CrossRefGoogle Scholar
  9. Bukowski TC, Linn CD, Christenson TR (2001) Copulation and sperm release in Gasteracantha cancriformis (Araneae: Araneidae): differential male behaviour based on female mating history. Anim Behav 62:887–895CrossRefGoogle Scholar
  10. Calbacho-Rosa L, Peretti AV (2015) Copulatory and post-copulatory sexual selection in haplogyne spiders, with enphasis on pholcidae and oonopidae. In: Peretti AV, Aisenberg A (eds) Cryptic female choice in arthropods: patterns, mechanisms and prospects. Springer, New York, pp 109–137CrossRefGoogle Scholar
  11. Calbacho-Rosa L, Córdoba-Aguilar A, Peretti AV (2010) Occurrence and duration of post-copulatory mate guarding in a spider with last sperm precedence. Behaviour 147:1267–1283CrossRefGoogle Scholar
  12. Calbacho-Rosa L, Galicia-Mendoza I, Dutto MS, Córdoba-Aguilar A, Peretti AV (2013) Copulatory behavior in a pholcid spider: males use specialized genitalic movements for sperm removal and copulatory courtship. Naturwissenschaften 100:407–416CrossRefGoogle Scholar
  13. Chapman T, Davies SJ (2004) Functions and analysis of the seminal fluid proteins of male Drosophila melanogaster fruit flies. Peptides 25:1477–1490CrossRefGoogle Scholar
  14. Cordero C (1995) Ejaculate substances that affect female insect reproductive physiology and behavior: honest or arbitrary traits? J Theor Biol 174:453–461CrossRefGoogle Scholar
  15. Cordero-Rivera A (2017) Sexual conflict and the evolution of genitalia: male damselflies remove more sperm when mating with a heterospecific female. Sci Rep 7:7844CrossRefGoogle Scholar
  16. Costa FG (1979) Análisis de la cópula y de la actividad postcopulatoria de Lycosa malitiosa Tullgren (Araneae, Lycosidae). Rev Bras Biol 39:361–376Google Scholar
  17. Costa FG, Toscano-Gadea CA (2003) Experimental interruption and re-initiation of mating in a wolf spider: an analysis of behavioural patterns and resultant progeny. Ethol Ecol Evol 15:173–181CrossRefGoogle Scholar
  18. Danielson-François AM (2006) Natural history of Glenognatha emertoni (Araneae, Tetragnathidae): mating behavior and sperm release in a haplogyne. J Arachnol 34:387–398CrossRefGoogle Scholar
  19. Danielson-François AM, Bukowski TC (2005) Female mating history influences copulation behavior but not sperm release in the orb-weaving spider Tetragnatha versicolor (Araneae, Tetragnathidae). J Insect Behav 18:131–148CrossRefGoogle Scholar
  20. Dickinson JL (1986) Prolonged mating in the milkweed leaf beetle Labidomera clivicollis (Coleoptera: Chrysomelidae): a test of “sperm-loading” hypothesis. Behav Ecol Sociobiol 18:331–338CrossRefGoogle Scholar
  21. Eberhard WG (1991) Copulatory courtship and cryptic female choice in insects. Biol Rev 66:1–31CrossRefGoogle Scholar
  22. Eberhard WG (1994) Evidence for widespread courtship during copulation in 131 species of insects and spiders, and implications for cryptic female choice. Evolution 48:711–733CrossRefGoogle Scholar
  23. Eberhard WG (1996) Female control: sexual selection by cryptic female choice. Princeton University Press, PrincetonGoogle Scholar
  24. Eberhard WG (2004) Why study spider sex: special traits of spiders facilitate studies of sperm competition and cryptic female choice. J Arachnol 32:545–556CrossRefGoogle Scholar
  25. Eberhard WG (2010) Rapid divergent evolution of genitalia: therory and data updated. In: Leonard JL, Cordoba-Aguilar A (eds) The evolution of primary sexual characters in animals. Oxford University Press, New York, pp 40–78Google Scholar
  26. Eberhard WG, Huber BA (2010) Spider genitalia: precise maneuvers with a numb structure in a complex lock. In: Leonard JL, Cordoba-Aguilar A (eds) The evolution of primary sexual characters in animals. Oxford University Press, New York, pp 249–284Google Scholar
  27. Elgar MA (1995) The duration of copulation in spider: comparative patterns. Rec West Aust Mus Suppl 52:1–11Google Scholar
  28. Foelix RF (2011) Biology of spiders, 3rd edn. Oxford Univerity Press, New YorkGoogle Scholar
  29. Gabel E, Uhl G (2013) How to prepare spider sperm for quantification. Arachnology 16:109–112CrossRefGoogle Scholar
  30. Herberstein ME, Wignall A (2011) Introduction: spider biology. In: Herberstein ME (ed) Spider behaviour, flexibility and versatility. Cambridge Univerity Press, New York, pp 1–21CrossRefGoogle Scholar
  31. Hosken DJ, Stockley P (2004) Sexual selection and genital evolution. Trends Ecol Evol 19:87–93CrossRefGoogle Scholar
  32. Huber BA (1995) Copulatory mechanism in Holocnemus pluchei and Pholcus opilionoides, with notes on male cheliceral apophyses and stridulatory organs in Pholcidae (Araneae). Acta Zool (Stockh) 76:291–300CrossRefGoogle Scholar
  33. Huber BA (2000) New World pholcid spiders (Araneae: Pholcidae): a revision at generic level. Bull Am Mus Nat Hist 254:1–348CrossRefGoogle Scholar
  34. Huber BA (2014) Pholcidae. In: Claps LE, Morrone JJ (eds) Roig-Juñet S. Biodiversidad de artropodos argentinos Editorial INSUE-UNT, San Miguel de Tucumán, pp 131–141Google Scholar
  35. Huber BA, Eberhard WG (1997) Courtship, copulation, and genital mechanics in Physocyclus globosus (Araneae, Pholcidae). Can J Zool 74:905–918CrossRefGoogle Scholar
  36. Jakob EM (1991) Costs and benefits of group living for pholcid spiderlings: losing food, saving silk. Anim Behav 48:482–484Google Scholar
  37. Kamimura Y (2005) Last-male paternity of Euborellia plebeja, an earwig with elongated genitalia and sperm-removal behavior. J Ethol 23:35–41CrossRefGoogle Scholar
  38. Kaster JL, Jakob EM (1997) Last-male sperm priority in a haplogyne spider (Araneae: Pholcidae): correlations between female morphology and patterns of sperm usage. Ann Entomol Soc Am 90:254–259CrossRefGoogle Scholar
  39. Kelly DA, Moore BC (2016) The morphological diversity of intromittent organs: an introduction to the symposium. Integr Comp Biol 56:630–634CrossRefGoogle Scholar
  40. Laborda A, Simó M (2008) First south American records of Holocnemus pluchei (Scopoli, 1763) and Spermophora senoculata (Duges, 1836) (Araneae: Pholcidae). Ganaya 72:261–265Google Scholar
  41. Leonard J, Córdoba-Aguilar A (2010) The evolution of primary sexual characters in animals. Oxford University Press, New YorkGoogle Scholar
  42. Linn CD, Molina Y, Diffata J, Christenson TE (2007) The adaptive advantage of prolonged mating: a test of alternative hypotheses. Anim Behav 74:481–485CrossRefGoogle Scholar
  43. Parker GA, Simmons LW, Kirk H (1990) Analyzing sperm competition data: simple models for predicting mechanisms. Behav Ecol Sociobiol 27:55–65CrossRefGoogle Scholar
  44. Peretti AV, Aisenberg A (2015) Cryptic female choice in arthropods: patterns, mechanisms and prospects. Springer, New YorkCrossRefGoogle Scholar
  45. Peretti AV, Eberhard WG (2010) Cryptic female choice via sperm dumping favours male copulatory courtship in a spider. J Evol Biol 23:271–281CrossRefGoogle Scholar
  46. Peretti AV, Eberhard WG, Briceño RD (2006) Copulatory dialogue: female spiders sing during copulation to influence male genitalic movements. Anim Behav 72:413–421CrossRefGoogle Scholar
  47. Porter AH, Jakob EM (1990) Allozime variation in the introduced spider Holocnemus pluchei (Araneae: Pholcidae) in California. J Arachnol 18:313–319Google Scholar
  48. R Core Team (2016) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL
  49. Schäfer MA, Uhl G (2002) Determinants of paternity success in the Pholcus phalangioides (Pholcidae, Araneae): the role of male and female mating behaviour. Behav Ecol Sociobiol 51:368–377CrossRefGoogle Scholar
  50. Schneider JM, Gilberg S, Fromhage L, Uhl G (2006) Sexual conflict over copulation duration in a cannibalistic spider. Anim Behav 71:781–788CrossRefGoogle Scholar
  51. Schöfl G, Taborsky M (2002) Prolonged tandem formation in firebugs (Pyrrhocoris apterus) serves mate-guarding. Behav Ecol Sociobiol 52:426–433CrossRefGoogle Scholar
  52. Simmons LW (2001) Sperm competition and its evolutionary consequences in the insects. Princeton University Press, New JerseyGoogle Scholar
  53. Simmons LW (2014) Sexual selection and genital evolution. Austral Entomol 53:1–17CrossRefGoogle Scholar
  54. Siva-Jothy MT (1987) The structure and function of the female sperm-storage organs in libellulid dragonflies. J Insect Physiol 33:559–567CrossRefGoogle Scholar
  55. Siva-Jothy MT, Tsubaki Y (1989) Variation in copulation duration in Mnais pruinosa pruinosa Selys (Odonata: Calopterygidae), I: alternative mate-securing tactics and sperm precedence. Behav Ecol Sociobiol 24:39–45CrossRefGoogle Scholar
  56. Snow LS, Andrade MC (2004) Pattern of sperm transfer in redback spiders: implications for sperm competition and male sacrifice. Behav Ecol 15(5):785–792CrossRefGoogle Scholar
  57. Szirányi A, Kiss B, Samu F, Harand H (2005) The function of long copulation in the wolf spider Pardosa agrestis (Araneae, Lycosidae) investigated in a controlled copulation duration experiment. J Arachnol 33:408–414CrossRefGoogle Scholar
  58. Uhl G (1994) Genital morphology and sperm storage in Pholcus phalangioides (Fuesslin, 1775) (Pholcidae, Aranae). Acta Zool 75:1–12CrossRefGoogle Scholar
  59. Uhl G (1996) Sperm storage secretion of female cellar spiders (Pholcus phalangioides; Araneae): a gel-electrophoretic analysis. J Zool 240:153–161CrossRefGoogle Scholar
  60. Uhl G, Nessler SH, Schneider JM (2010) Securing paternity in spiders? A review on occurrence and effects of mating plugs and male genital mutilation. Genetica 138:75–104CrossRefGoogle Scholar
  61. Uhl G, Kunz K, Vöcking O, Lipke E (2014) A spider mating plug: origin and constraints of production. Biol J Linn Soc 113:345–354CrossRefGoogle Scholar
  62. Weldingh DL, Toft S, Larsen ON (2011) Mating duration and sperm precedence in the spider Linyphia triangularis. J Ethol 29:143–152CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Facultad de Ciencias Exactas, Físicas y Naturales, Departamento de Diversidad Biológica y EcologíaUniversidad Nacional de CórdobaCórdobaArgentina
  2. 2.Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Laboratorio de Biología Reproductiva y EvoluciónInstituto de Diversidad y Ecología Animal (IDEA)CórdobaArgentina
  3. 3.Departamento de Ecología Evolutiva, Instituto de EcologíaUniversidad Nacional Autónoma de MéxicoDistrito FederalMexico

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