Recent studies have found that Gopher Tortoise, Gopherus polyphemus, populations in southern Mississippi exhibit low recruitment, due in part to very low hatching success of their eggs. We sought to determine if the cause(s) of this low hatching success was related to egg quality (intrinsic factors), unsuitability of the nest environment (extrinsic factors), or a combination of the two. In 2003, hatching success was monitored simultaneously for eggs from the same clutches that were incubated in the laboratory and left to incubate in nests. A subset of randomly chosen eggs from each clutch was incubated in the laboratory under physical conditions that were known to be conducive to successful hatching to estimate the proportion of eggs that were capable of hatching in a controlled setting. Hatching success in the laboratory was compared with that of eggs incubated in natural nests to estimate the proportion of eggs that failed to hatch presumably from extrinsic factors. Laboratory hatching success was 58.8%, suggesting that roughly 40% of the eggs were intrinsically incapable of hatching even when incubated under controlled conditions. Hatching success in natural nests, 16.7%, was significantly lower than hatching success in the laboratory, suggesting that approximately 42.1% of eggs were capable of hatching but failed to hatch due to some extrinsic aspect(s) of the nest environment. Thus, the low hatching success of Gopher Tortoise eggs in southern Mississippi appears to be attributable to a combination of intrinsic (egg quality) and extrinsic (nest environment) factors.