Influence of mating order on courtship displays and stored sperm utilization in Hermann's tortoises (Testudo hermanni hermanni)
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Sperm storage is widespread in many vertebrate groups, and it is frequently associated with promiscuous mating systems. Chelonian species are one of the most outstanding examples of a promiscuous group capable of long-term sperm storage; specialized structures have evolved within the oviducts of these vertebrates to ensure sperm vitality across reproductive cycles. Thus far, few studies have investigated the factors regulating multiple paternity, sperm usage by females and paternity distribution in successive clutches. This study aimed to investigate the effect of mating order on male mounting and reproductive success in Testudo hermanni hermanni, combining behavioural and genetic data. A series of planned matings were performed, within which experimental females were mated sequentially to two different males under controlled conditions. Observations conducted during experimental matings revealed that courtship displays did not significantly differ between the first and second males to mate with a female. Interactions ending with a mount were characterized by a significantly higher intensity and occurrence of determinate courtship behaviours, for example biting and running after the female. Paternity analysis performed on hatchlings produced from experimental females revealed that 46 % of the clutches were multi-sired. A significant contribution of previous years' partners was still found, thus confirming the long-term sperm storage within the female oviduct in this species. Finally, mating order did not significantly affect the reproductive success of experimental males during the on-going reproductive season.
KeywordsTestudo hermanni Mating success Sperm storage Female promiscuity Paternity distribution
Many thanks are due to Sandra Cardoso for her precious help and support in our research. We also thank Giulia Santalmasi and Dania Benini for their work in the field and in the laboratory. We also express our warm thanks to Thomas Madsen, an associate editor of Behavioral Ecology and Sociobiology, for his precious comments and suggestions and the accurate revision of our manuscript: this final version of our manuscript greatly benefitted from his invaluable help. Finally we thank Jenny Booth for the linguistic revision of the manuscript. This study was financed by the funds of Associazione TartaEtruria and by the funds ex-60 % of Prof. Marco Vannini and Dott. Stefano Cannicci.
All the experiments performed comply with the Italian and the European current laws on Testudo species.
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