Majelantle, Tshepiso LesediGanswindt, AndreGanswindt, Stefanie BirgitHagenah, NicoleHart, Daniel WilliamBennett, Nigel Charles2023-11-132023-11-132023-09Majelantle, T.L.; Ganswindt, A.; Ganswindt, S.B. et al. Non-Invasive Quantification of Faecal and Urine Reproductive Hormone Metabolites in the Naked Mole-Rat (Heterocephalus glaber). Animals 2023, 13, 3039. https://doi.org/10.3390/ani13193039.2076-2615(online)10.3390/ani13193039http://hdl.handle.net/2263/93247DATA AVAILABILITY : The data generated and subsequently analysed during this study will be sent by the corresponding author upon request.SUPPLEMENTARY MATERIALS : FIGURE S1: Serial dilutions of faecal extracts for the 5α-Progesterone EIA. FIGURE S2: Serial dilutions of faecal extracts for the Epiandrosterone EIA. TABLE S1: Details on EIA antibodies, label, standard, and references for each EIA used in the study.The naked mole-rat (Heterocephalus glaber) occurs in colonies with a distinct dominance hierarchy, including one dominant, breeding female (the queen), 1–3 breeding males, and non-reproductive subordinates of both sexes that are reproductively suppressed while in the colony. To non-invasively evaluate reproductive capacity in the species, we first had to examine the suitability of enzyme immunoassays (EIAs) for determining progestogen and androgen metabolite concentrations in the naked mole-rat, using urine and faeces. A saline control and gonadotrophin-releasing hormone (GnRH) were administered to twelve (six males and six females) naked mole-rats which were previously identified as dispersers and housed singly. The results revealed that urine is possibly not an ideal matrix for progestogen and androgen metabolite quantification in naked mole-rats as no signal was detected in the matrix post GnRH administration. A 5α-Progesterone EIA and an Epiandrosterone EIA were identified as suitable for quantifying faecal progesterone metabolites (fPMs) and faecal androgen metabolites (fAMs) in males and females, respectively. The results suggest that there are individual variations in baseline fPM and fAM concentrations, and only two out of six females and no males exhibited an increase in fPM concentrations greater than 100% (−20% SD) post GnRH administration. Conversely, only four out of six females and three out of six males had an increase in fAM concentrations greater than 100% (−20% SD) following GnRH administration. These results imply that some naked mole-rat individuals have a reduced reproductive capacity even when they are separated from the queen.en© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).GnRH administrationFaecal androgen metabolitesFaecal progestogen metabolitesUrine androgen metabolitesNaked mole-rat (Heterocephalus glaber)SDG-03: Good health and well-beingEnzyme immunoassay (EIA)Urine progestogen metabolitesAfrican mole-rats (Bathyergidae)Article