Non-Invasive Reproductive Hormone Monitoring in Endangered Pygmy Hog (Porcula salvania)

The Pygmy hog (Porcula Salvania), till recently, classified as a critically endangered suid, is facing the threat of extinction globally due to habitat degradation. Efforts are being made to protect the pygmy hogs from extinction and breed them in captivity under Pygmy Hog Conservation Pro-gramme (PHCP). However, very little information is available on the reproductive physiology of pygmy hogs. Therefore, the present study aimed to standardize enzyme immunoassays (EIAs) for monitoring pregnancy and reproductive status using progesterone and testosterone metabolites. A total of 785 faecal samples were collected from five females and two males over a period of one year from PHCP Research and Breeding Centre, Guwahati, Assam. High-pressure liquid chro-matography (HPLC) analysis revealed the presence of immunoreactive progesterone and testos-terone metabolites in faeces. Mating was observed in all the five females and four of them gave birth successfully. We were able to detect pregnancy using faecal progesterone metabolites. Based on mating and parturition, the mean gestation period was estimated to be 153.25 days from four females. The breeding centre recorded 172 births between 1996 and 2000 and found strong sea-sonality in births and most of the births were between May and June. Faecal testosterone metabo-lites were significantly higher in the breeding season than the non-breeding season. This is the first study and will help in future breeding programs in other captive breeding centres and reproduc-tive monitoring of reintroduced populations.

Measurement of Fecal Testosterone Metabolites in Mice: Replacement of Invasive Techniques

Testosterone is the main reproductive hormone in male vertebrates and conventional methods to measure testosterone rely on invasive blood sampling procedures. Here, we aimed to establish a non-invasive alternative by assessing testosterone metabolites (TMs) in fecal and urinary samples in mice. We performed a radiometabolism study to determine the effects of daytime and sex on the metabolism and excretion pattern of radiolabeled TMs. We performed physiological and biological validations of the applied EIA to measure TMs and assessed diurnal fluctuations in TM excretions in male and female mice and across strains. We found that males excreted significantly more radiolabeled TMs via the feces (59%) compared to females (49.5%). TM excretion patterns differed significantly between urinary and fecal samples and were affected by the daytime of ³H-testosterone injection. Overall, TM excretion occurred faster in urinary than fecal samples. Peak excretion of fecal TMs occurred after 8 h when animals received the 3H-testosterone in the morning, or after 4 h when they received the 3H-testosterone injection in the evening. Daytime had no effect on the formed TMs; however, males and females formed different types of TMs. As expected, males showed higher fecal TM levels than females. Males also showed diurnal fluctuations in their TM levels but we found no differences in the TM levels of C57BL/6J and B6D2F1 hybrid males. Finally, we successfully validated our applied EIA (measuring 17β-hydroxyandrostane) by showing that hCG (human chorionic gonadotropin) administration increased TM levels, whereas castration reduced them. In conclusion, our EIA proved suitable for measuring fecal TMs in mice. Our non-invasive method to assess fecal TMs can be widely used in various research disciplines like animal behavior, reproduction, animal welfare, ecology, conservation, and biomedicine.

Hens (Gallus gallus domesticus) with larger spurs have higher faecal testosterone levels; evidence of female to male - like phenotype

Spurs are a testosterone-dependent secondary sexual characteristic of male chickens (Gallus gallus domesticus) and are used as ‘weapons’ in intra-sexual fighting. While predominantly a male feature, they are also found in a small number of females. Here we show that faecal testosterone metabolites of adult hens (i.e. females) with large spurs are significantly greater than those of females with smaller spurs. The presence of spurs in hens together with elevated testosterone levels are indicative of a masculinisation process that may increase with age. This is of particular interest in the study of female to male sex-reversal in fowl.

Hens (Gallus gallus domesticus) with larger spurs have higher faecal testosterone levels; evidence of female to male - like phenotype

Spurs are a testosterone-dependent secondary sexual characteristic of male chickens (Gallus gallus domesticus) and are used as ‘weapons’ in intra-sexual fighting. While predominantly a male feature, they are also found in a small number of females. Here we show that faecal testosterone metabolites of adult hens (i.e. females) with large spurs are significantly greater than those of females with smaller spurs. The presence of spurs in hens together with elevated testosterone levels are indicative of a masculinisation process that may increase with age. This is of particular interest in the study of female to male sex-reversal in fowl.