Effects of Two Different Dietary Calcium Concentrations on Bone Density and Skin Microbiome in Lemur Tree Frogs (Agalychnis lemur)

The lemur tree frog (<i>Agalychnis lemur</i>), a critically endangered species, can benefit from ex situ conservation programs; however, managing amphibians under human care presents challenges, including the provision of appropriate nutrition. House crickets (<i>Acheta domesticus</i>), a common feeder insect, have an inverse calcium to phosphorus ratio (Ca:P; 0.15:1) and low calcium content (<0.3%). While gut-loading crickets with an 8% calcium diet can improve their calcium concentrations, no study has assessed the effects of dietary calcium on bone development in <i>Agalychnis</i> spp. Moreover, no study has examined how diet impacts the gut-skin axis and skin microbiome of these frogs. This study examined how crickets gut-loaded with either a 1.3% or 8% calcium diet affected lemur tree frog bone density and skin microbiome. We hypothesized that frogs consuming the 8% calcium diet would exhibit significantly higher Hounsfield units (HU; bone density) over time, as measured by micro-computed tomography (mCT), and that dietary calcium concentration would have no effect on skin bacterial and fungi microbiomes. Eleven juvenile lemur tree frogs underwent mCT scans at baseline and 90 and 180 days. Total body volume of interest analysis showed a significant increase in HU in the 8% calcium group compared to the 1.3% group (F = 9.9, <i>p</i> = 0.01). There was no significant difference noted in the alpha or beta diversities for the bacterial and fungal microbiomes between dietary groups. This study provides the first evidence of dietary calcium's impact on bone density in lemur tree frogs, offering valuable insights for improving ex situ management of this species.