Temperature effects on metabolism and energy requirement during the fast growth phase in the red-footed tortoise, Chelonoidis carbonaria.

Early life is a challenging phase because of the high rates of morphophysiological development and growth. Changes in ambient temperature, which directly affect energy metabolism and digestive functions in ectotherms, may be of great impact during this phase. We addressed this issue in red-footed tortoise (Chelonoidis carbonaria) hatchlings kept in captivity. To this end, we investigated the effect of temperature (28 °C and 18 °C) on mass-specific gross energy intake (GEIm), daily body mass gain (MG), daily intake of gross energy (GEI), digestible energy (DEI), resting metabolic rate (RMR), and specific dynamic action (SDA) components during different seasons in the first 13 months after hatching. Greater GEIm and MG were observed in spring (381.7 ± 84.9 J.g-0.86.day-1 and 0.9 ± 0.4 g.day-1) and summer (356.9 ± 58.9 J.g-0.86.day-1 and 1.0 ± 0.4 g.day-1). The highest and lowest RMRs at 28 °C were observed in spring (36.4 ± 5.1 kJ.kg-1.day-1) and winter (22.4 ± 6.2 kJ.kg-1.day-1), respectively. Regardless season, hatchlings showed greater GEI and DEI, O2 consumption, CO2 production, RMR, maximum metabolic rate after feeding (FMRMAX), and heat increment (FMRMAX- RMR) at 28 °C compared to 18 °C. In addition, the significant body mass influence showed allometric exponents of 0.62 at 28 °C and 0.92 at 18 °C for RMR. Our results indicate an important effect of environmental temperature on energy requirements and utilization in C. carbonaria hatchlings, which is seasonally influenced even in this early phase of life.

Starch and fiber intake effects on energy metabolism, growth, and carapacial scute pyramiding of red-footed tortoise hatchlings (Chelonoidis carbonaria).

Tortoise husbandry includes reports of excessive growth and carapace pyramiding, although triggers still remain to be fully elucidated. Juvenile red-footed tortoises (Chelonoidis carbonaria) were fed with two different diets, one high in fiber (HF; 14.2% crude fiber; 39.2% neutral detergent fiber, NDF; dry matter basis, DMB) and one high in starch (HS; 27.7% DMB), to assess effects on energy metabolism, nutrient digestibility, and growth. A total of 20 hatchlings (10 per diet) were used to evaluate: apparent digestibility coefficients (Da) of nutrients and gross energy (GE), passage times at 5 and 11 months of age; resting and post-prandial metabolic rates at 6 and 12 months of age; growth rates; pyramiding; and estimated body composition. Animals fed HS showed higher mass-specific intake of digestible energy (113.9 ± 32.1 kJ kg-1 day-1 vs. 99.6 ± 35.3 kJ kg-1 day-1; P < 0.05), digestible DM (6.1 ± 1.8 g kg-1 day-1 vs. 5.0 ± 1.8 g kg-1 day-1; P < 0.01), shorter transit (3 ± 1 days vs. 4 ± 1 days; P < 0.01) and retention times (8 ± 2 days vs. 10 ± 2 days; P < 0.01), and higher Da of DM, starch, NDF, and GE. Crude protein Da was higher for HF. Rest and post-prandial metabolic rates, and pyramiding degree were not affected by diets. At 13 months, the animals from HS presented wider plastrons and carapaces, and higher carapace width growth rates. In addition, these animals had lower body mineral content (1.88 ± 0.15% vs. 2.15 ± 0.19%; P < 0.01) and bone density (0.13 ± 0.01 g mm-2 vs. 0.15 ± 0.02 g mm-2; P < 0.02). Results provide evidence that highly digestible foods can accelerate shell growth and lower mineralization in this species.

Influence of incubation temperature on embryo development, hatchling morphology and early growth rate in red-footed tortoise (Chelonoidis carbonaria).

Reptile embryos respond to temperature changes with metabolic and physiological adjustments that influence hatchling success, phenotype, behaviour, and growth rate. Climate change and global warming can affect the reptile population by altering the frequencies of hatchling survival and phenotypes. Therefore, previous studies proposed artificial incubation as a potential strategy for mitigating these effects. Red-footed tortoise (Chelonoidis carbonaria) eggs were collected and incubated at constant temperatures of 27.5 °C and 29.5 °C to investigate the physiological effects of temperature on embryo development, hatchling morphology, and early post-hatch growth rate. The direct effect of temperature on the incubation period, egg mass loss, hatching success, hatchling size, and mass was evaluated at hatching and three months of age. Hatchlings from 29.5 °C presented a shorter incubation period (141 days) than those from 27.5 °C (201 days; p < 0.05). Egg mass loss, hatchling mass, and size at hatching were not different between the incubation temperatures (p > 0.05). However, the hatching success (survival rate) was lower (64.5% versus 100%) in eggs incubated at 29.5 °C, but the hatchling mass and straight plastron width were higher at three months of age than those from eggs incubated at 27.5 °C (p < 0.05). These results indicate that incubation temperature influences hatching success and hatchling size and mass in the first months by influencing the early growth rate.

Copper deficiency-induced anemia is caused by a mitochondrial metabolic reprograming in erythropoietic cells.

The lack of copper has been associated with anemia, myelodysplastic syndromes and leukemia as well as with a loss in complex IV activity and an enlarged mitochondrial morphology. Mitochondria play a key role during the differentiation of hematopoietic stem cells by regulating the passage from a glycolytic to oxidative metabolism. The former is associated with cell proliferation and the latter with cell differentiation. Oxidative metabolism, which occurs inside mitochondria, is sustained by the respiratory chain, where complex IV is copper-dependent. We have hypothesized that a copper deficiency induces a mitochondrial metabolic reprogramming, favoring cell expansion over cell differentiation in erythropoiesis. Erythroid progression analysis of the bone marrow of mice fed with a copper deficient diet and of the in vitro erythropoiesis of human CD34+ cells treated with a bathocuproine - a copper chelator - showed a major expansion of progenitor cells and a decreased differentiation. Under copper deficiency, mitochondria switched to a higher membrane potential, lower oxygen consumption rate and lower ROS levels as compared with control cells. In addition, mitochondrial biomass was increased and an up-regulation of the mitochondrial fusion protein mitofusin 2 was observed. Most copper-deficient phenotypes were mimicked by the pharmacological inhibition of complex IV with azide. We concluded that copper deficiency induced a mitochondrial metabolic reprogramming, making hematopoietic stem cells favor progenitor cell expansion over cell differentiation.