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Annexe III _____________________________________________ corresponding alternatively to the beginning and the middle of each third of the total incubation time minus the first 5 days. The three periods of incubation were determined as the totally duration of incubation (estimated approximately at 60 days at this temperature), divided in three equal periods of time. We defined three water potentials: (i) a “wet substrate” (-10 KPa; i.e. 2 g of sterilized water/ g of vermiculite) that represents a soil after rainfalls, (ii) an “intermediate substrate” (-398 KPa; i.e. 0.44 g of sterilized water / g of vermiculite) and (iii) a “dry substrate” (-1000 KPa; i.e. 0.09 g of sterilized water/ g of vermiculite) that represents a soil in drought period. At each period of incubation, two groups of three different and new boxes were transferred from intermediate to the wet and to the dry substrates (see Table 1). In the middle of the period, the two groups of boxes were transferred again with, for each group, one box staying in the same substrate and the two others changed for the two other substrates. One box stayed on intermediate substrate during all the course of incubation. Finally, each box represented a different treatment of moisture substrate and has undergone a maximum of two transfers during one period of incubation. Data collection during incubation Every other day, from day 5 until hatching, eggs were individually weighed with a Sartorius electronic balance (to the nearest 0.001g). Boxes without eggs were also weighed to adjust water potential in replacing water absorbed by eggs or escaped around edges of the box sides. Eggs covered by Fungus or other aborted eggs were removed and opened to check the content. Eggs with no viable embryos were excluded from statistical analyses of change in egg mass during incubation. After piping and before hatching, each turtle was placed on wet cotton in an individual plastic cup. Incubation time of each egg was defined as the pip date minus the date where a white spot appeared on the eggshell. 6
Annexe III _____________________________________________ Husbandry, hatchling size and sex identification After emergence, turtles were maintained in the humidified cup until total absorption of yolk. Hatchlings were then weighed to the nearest 0.001 g on an electronic Sartorius balance and individually marked by shell notching. Approximately 15-20 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. During 15 months, hatchlings were weighted approximately every two months. After this period, all juveniles were sacrified and sex was determined a posteriori by dissection and microscopic observation of gonad morphology (Pieau, 1972; Yntema, 1976). Juveniles with an indeterminate sex phenotype (n=12) were removed from statistical analyses. Statistical analyses All data were tested for normality (Kolmogorov-Smirnov) and for homogeneity (Bartlett’s test) prior to further statistical analysis. We used a mixed linear model to analyze normally distributed data and a generalized mixed model for not normally distributed data. Statistical analyses were performed using SAS for windows version 9.1 (SAS institute Inc., Cary, NC, USA, 2003). 7
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N° d’ordre : UNIVERSITE PARIS-SU
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Remerciements Je remercie tout d’
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Sommaire __________________________
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Introduction ______________________
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Introduction ______________________
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Introduction ______________________
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Introduction ______________________
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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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III - Etudes expérimentales des pr
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Conclusion & Perspectives _________
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Bibliographie _____________________
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Bibliographie _____________________
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Bibliographie _____________________
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Bibliographie _____________________
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Page 151 and 152: TABLE DES ANNEXES A. Supplementary
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TURTLE, AN EXOTIC AND INTRODUCED SP
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