Sperm storage and low incidence of multiple paternity in the European pond turtle, Emys orbicularis: A secure but costly strategy?

The freshwater pond turtle, Emys orbicularis, has recently suffered from population declines throughout its range, mainly due to habitat destruction. The mating strategies of this species were studied using genetic data from successive clutches within and between years. To test for the occurrence and frequency of multiple paternity and sperm storage, genetic paternity at six microsatellite markers was assessed in 114 embryos and hatchlings from single and subsequent clutches of 11 females (including clutches from the same or consecutive years). Multiple paternity was rare and only found in two out of 20 clutches from 11 females. All annual successive clutches and 58% of the clutches in the next year, were fertilized with sperm from the same male. The use of stored sperm is thus a frequent strategy in E. orbicularis. However, hatching rate, hatchling mass, and hatchling length decreased in clutches fertilized by stored sperm, suggesting sperm depletion or deterioration through time. The occurrence of stored sperm despite an associated reduced reproductive output indicated that mating and/or the fertilization process is costly to females. The low incidence of multiple paternity may simply be the residual consequence of the capacity to store viable sperm. These results provide important and innovative insights for the conservation of E. orbicularis. In threatened populations, management strategies may aim to enhance effective copulations in order to increase the reproductive output of females.