Optimizing captive short‐beaked echidna (Tachyglossus aculeatus) fecal sample identification and hormonal analysis

Abstract

The objectives of this study were to develop a fecal marking protocol to distinguish male from female samples during the echidna breeding season and to determine if normalizing fecal progesterone metabolite data for inorganic content improves the detection of biologically relevant changes in metabolite concentrations. Over a period of 6 weeks, four echidnas were provided with green food coloring powder mixed into 20 g of their regular feed with the dose adjusted weekly by 0.05 g. The proportion of organic (feces) versus inorganic matter (sand) in the fecal samples of three echidnas was determined by combustion of organic matter. Hormonal data was then expressed as metabolite concentration per total dry mass (with sand) of extracted sample versus metabolite concentration per total mass of organic material (without sand). The optimal dose of food coloring powder was 0.30 g: this was excreted in the feces of all echidnas within 24 h of consumption with color present for two consecutive days.

Gut microbiota in the short‐beaked echidna (Tachyglossus Aculeatus) shows stability across gestation

“…The timing of these events (e.g., copulation, egg‐laying) was paired with existing literature (Dutton‐Regester et al, 2021; Dutton‐Regester, Roser, Meer, Russell, et al, 2023; Morrow et al, 2009) to determine what stage of gestation each female echidna was in on that collection date. To further discriminate fecal samples during copulation events when both male and female echidnas were present in the same enclosure, males were provided with Chromacake green water‐soluble food coloring powder (Baking Pleasures) in 20 g of their food portion (Dutton‐Regester, Roser, Meer, Renfree, et al, 2023; Dutton‐Regester, Roser, Meer, Russell, et al, 2023). While each enclosure was searched daily for fecal samples, a sample was not necessarily always found; this was likely due to a lack of defecation during the preceding 24 h. Where fecal material was present, samples were placed into labeled plastic Ziplock bags and stored at −20°C for later processing.…”

Faecal microbiota in the short-beaked echidna (Tachyglossus aculeatus) shows stability across gestation

“…The timing of these events (e.g., copulation, egg-laying) was paired with existing literature [8, 9,47] to determine what stage of gestation each female echidna was in on that collection date. To further discriminate faecal samples during copulation events when both male and female echidnas were present in the same enclosure, males were provided with Chromacake green water-soluble food coloring powder (Baking Pleasures, Queensland, Australia) in 20 g of their food portion [9,53]. While each enclosure was searched daily for faecal samples, a sample was not necessarily always found; this was likely due to a lack of defecation during the preceding 24 h. Where faecal material was present, samples were placed into labelled plastic Ziplock bags and stored at − 20°C for later processing.…”