**5. Conclusions**

This work represents a primer in giving quantitative information about the urinary metabolome of captive gira ffes, as detected by untargeted 1H-NMR. Foraging behaviors and daily activity could be considered as one of the main reasons for the di fferences we highlighted that are linked to sex and age. A preliminary observation conducted on two female gira ffes suggests that 1H-NMR based metabolomics could be conveniently applied to monitor modifications occurring during pregnancy, some of which are potentially related to inflammatory status triggered by modification of the microbiota profile.

**Supplementary Materials:** The following are available online at http://www.mdpi.com/2218-1989/10/4/157/s1, Table S1: Concentration (mmol/L, median (IQR)) of the molecules quantified by 1H-NMR in all the samples studied in the present investigation, sorted by abundance. Table S2. Concentration (mmol/L) of the molecules quantified by 1H-NMR in the samples collected during and after pregnancy.

**Author Contributions:** C.Z., L.L., S.F., and G.I. conceived and designed the research. C.Z. and L.L. performed metabolomics analysis. S.F. collected the samples. C.Z., L.L., S.F., and G.I. wrote the manuscript. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research received no external funding.

**Acknowledgments:** Chenglin Zhu gratefully acknowledges financial support from Chinese Scholarship Council (grant n◦ 201606910076). Part of the samples used in this study was collected with the financial Grant won by Sabrina Fasoli and donated by the Zebra Foundation for Veterinary Zoological Education in 2019. The authors would like to thank Bandoli Francesca, Cordon Rossana, Cotignoli Chiara, Laguardia Daniele, Laricchiuta Pietro, Sandri Camillo, Schneider Rainer, and Spiezio Caterina for their support. The authors also would like to thank all staff of the zoos involved in this study for their precious help during the sampling and Vito Barnaba, Pietro Ciaccia, Giacomo Melani, and Fulvio Pendezza for their fundamental assistance.

**Conflicts of Interest:** The authors declare no conflict of interest.
