Temperature-Dependent Sex Determination and Artificial Incubation of Tuatara, Sphenodon Punctatus
Authors:   , Nelson Nicola J 
Published: November 2021

<p>Juveniles resulting from artificially induced and incubated eggs are often used to found or augment populations of rare reptiles, but both procedures may compromise the health of hatchlings or their fitness in natural environments. I aimed to test whether these procedures affected size or performance of juvenile tuatara, Sphenodon punctatus, New Zealand reptiles with temperature-dependent sex determination (TSD). Size and performance are phenotypic traits likely to influence fitness and eventual lifetime reproductive success, and are thus important measures of the suitability of artificial induction and incubation techniques for conservation management. I incubated 320 tuatara eggs artificially at 18, 21 and 22ºC; 52% of these were obtained by induction, the remainder were collected from natural nests. An additional 25 natural nests were left intact for investigation of TSD and effects of incubation temperature in nature. Juveniles from all incubation regimes were kept for ten months post-hatching in similar rearing conditions and sexed by laparoscopy. Induced eggs were significantly smaller than naturally laid eggs, and resulted in significantly smaller hatchlings, even when variation among clutches was accounted for. Incubation temperature did not greatly influence size at hatching, but was an important determinant of size by ten months of age; initial egg mass was the most important factor affecting size of hatchlings. Data indicate that TSD occurs in nature. The sex of hatchlings from 21 nests was investigated: 10 nests produced 100% male hatchlings, 4 nests produced 100% female hatchlings, and only 7 nests produced mixed sex ratios which ranged from 11% to 88% males. Sex of juveniles was related to temperature with a larger proportion of males produced in warmer nests. The overall percentage of male hatchlings in natural nests was 64%. Hatching success was 65% from natural nests during the 1998/99 season. Incubation temperatures throughout the year ranged from 2.9 to 34.4ºC. Global warming is likely to skew the hatchling sex ratio towards males if female tuatara are unable to select nest sites according to environmental cues. Evidence from size patterns of tuatara incubated in natural nests supports differential fitness models for the adaptive significance of TSD. The evaluation of artificial incubation as a conservation management tool demonstrated that it is a procedure that benefits conservation as it can be used reliably to produce founders; hatching success was 94% during this study. The sex ratio of artificially incubated juveniles can be easily manipulated; the pivotal temperature lies between 21 and 22ºC. Constant artificial incubation conditions resulted in larger juveniles by ten months of age than those from natural incubation. Naturally incubated juvenile tuatara, however, were faster for their size, their reaction norm to predator stimuli was to run, and they were possibly more aggressive, suggesting naturally incubated juveniles could survive better in nature. No firm conclusions can be reached on the quality of artificially incubated juvenile tuatara because further research will be required to establish the relevance of performance test results in nature and consequences of incubation regimes in the longer term with respect to relative fitness of individuals.</p>