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

## TL;DR

This study shows that feeding lemur tree frogs crickets with higher calcium improves their bone density, though it doesn't affect their skin microbiome.

## Contribution

The study is the first to demonstrate the impact of dietary calcium on bone density in lemur tree frogs.

## Key findings

- Frogs fed high-calcium crickets had significantly higher bone density compared to those fed low-calcium crickets.
- There was no significant difference in skin microbiome diversity between the two dietary groups.
- This is the first study to assess dietary calcium's effect on bone development in Agalychnis spp.

## Abstract

Threatened species, such as lemur tree frogs (Agalychnis lemur), benefit from conservation programs. However, maintaining amphibians under human care presents challenges, including providing appropriate nutrition. Many frogs under human care are fed crickets, but these insects are naturally low in calcium. This study examined whether gut-loading crickets with a high-calcium diet could improve lemur tree frog bone density and affect bacterial and fungal skin communities (microbiome). There was no significant difference in the skin microbiome between groups; however, frogs offered high-calcium crickets had significantly higher bone density compared to frogs offered low-calcium crickets. These findings emphasize the importance of proper nutrition in protecting endangered frogs in captivity. Enhancing dietary calcium can help zoos and conservation programs raise healthier frogs, increasing their chances of survival and successful breeding. This research provides valuable insights for global amphibian conservation efforts by ensuring frogs under human care receive the necessary nutrition to thrive.

The lemur tree frog (Agalychnis lemur), 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 (Acheta domesticus), 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 Agalychnis 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, p = 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.