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Annexe IV _____________________________________________ cumbersome (20-25 cm carapace length), and most of them were released by unsuspecting owners in natural ponds. Thus, several reception centres have been created in France, particularly in the South of France, to fetch and take care of these exotic animals. To date, in the field, production of viable hatchlings of this species was only observed in the South of France. Females lay their eggs from June to July and hatchlings emerge from their nests in September (pers. com.). Hatchlings do not overwinter in the nest as commonly observed in natural populations of T. scripta elegans in northern America (Tucker & Paukstis, 1999). Egg collection Sixty gravid females of Trachemys scripta elegans were captured in artificial ponds in the South of France, in two zoological facilities located in the “Ferme aux Crocodiles” (Pierrelatte, France) and in the “Association Tortues Passion” (Vergèze, France). These two facilities are geographically very close to each other, and turtles present there come from everywhere in France, thus we considered the origin of turtles as random. Moreover, densities of animals in these two artificial ponds are approximately the same and turtles are fed twice a week with the same food. We decided thus to consider all turtles as a single population. Each captured female was isolated in a plastic box with some water. Oviposition was induced by injection of 1ml intra muscular oxytocin (Ewert & Legler, 1978). Each oviposited egg was identified by a code number on the eggshell for individual identification, weighed and placed on wet cotton in a cool box to prevent desiccation during transport to the laboratory. After egg deposition, adult females were released in their artificial ponds. Incubation of eggs at laboratory Fertile eggs were 2/3 buried in moistened vermiculite under controlled and intermediate water potential (-398 Kpa i.e. 0.44 g of sterilized water / g of vermiculite), and incubated in four 7
Annexe IV _____________________________________________ programmable incubators (Memmert, IPP 200-400; Schwabach, Germany). A first set of 190 fertilized eggs was incubated at a constant temperature of 28.9°C (regime A, Figure 1) that yields both sexes (Mrosovsky & Pieau, 1991; Godfrey, Delmas & Girondot, 2003). The other 112 eggs were shared and incubated at three different fluctuating temperatures (24 hourssinusoidal B, C and D regimes, Figure 1). Eggs were placed in covered plastic boxes, with a maximum of ten eggs per box. Eggs from the same clutch were dispersed as completely as possible among boxes within each temperature treatments (constant vs. fluctuant), in order to prevent confounding box and clutch effects. Twice a week, boxes were rotated to prevent local variations of temperature in incubators and weighed to adjust water potential. Husbandry After piping and before emerging, each hatchling turtle was placed on wet cotton in an individual plastic cup. After emergence, the turtles were maintained in the humidified cup until total absorption of yolk. Hatchlings were then weighed and individually marked by shell notching. Approximately 15 days after hatching, turtles were housed under standardized conditions in plastic tubs (60 cm long × 40 cm wide × 30 cm high) filled with 4 cm of water, in an air-conditioned room. Juveniles were randomly distributed among tubs, with a density of approximately 15 hatchlings per tub. Ambient temperature was maintained at 28°C, while water temperature varied between 22 and 23°C. Basking sites were placed in the tubs to allow thermoregulation. PowerSun UV lamps (Zoo Med laboratories Inc., San Luis Obispo, CA, USA) offered the appropriate intensity of UVB, UVA, heat and visible light with respect of photoperiod (10:14 L:D). Turtles were fed twice a week ad libitum with Hatchling Aquatic Turtle Food (Zoo Med laboratories Inc., San Luis Obispo, CA, USA). Tubs were cleaned two hours after each feeding. All juveniles were sacrificed when they were 16-month old for sexing. 8
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N° d’ordre : UNIVERSITE PARIS-SU
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Remerciements Je remercie tout d’
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I - La tortue à tempes rouges, un
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II - Les tortues à tempes rouges,
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TABLE DES ANNEXES A. Supplementary
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