Comparison of fecal storage methods for steroid analysis in black rhinoceroses (Diceros bicornis)
Publication Type: |
Journal Article |
Year of Publication: |
2004 |
Authors: |
Wieke T. Galama, Laura H. Graham, Anne Savage |
Publication/Journal: |
Zoo Biology |
Publisher: |
A Wiley Company, Inc., Wiley Subscription Services |
Keywords: |
enzyme immunoassay, fecal steroids, preservation of samples, progestagens |
ISBN: |
1098-2361 |
Abstract:
Abstract 10.1002/zoo.20017.abs Many field studies and conservation programs for wildlife species include noninvasive endocrine monitoring of gonadal function. Freezing fecal samples immediately after collection until further analysis is often not a viable option for researchers in remote areas. Phase 1 of this study was designed to compare different methods of preserving fecal samples over several time periods (30, 90, or 180 days) in order to determine which method provided the most accurate and reliable technique for measuring fecal progestagens. Fecal samples were collected from two female black rhinoceroses (Diceros bicornis) housed at Disney’s Animal Kingdom, Lake Buena Vista, FL. We compared three storage methods: 1) storing fecal samples without processing or preservatives (untreated), 2) storing an aliquot of fecal sample in 80% methanol (MeOH), and 3) drying the fecal sample in a solar box cooker prior to storage. Control samples (day 0) were collected and extracted, and then stored at −20°C until they were analyzed. Phase 2 of the study was designed to examine the effects of long-term storage (up to 180 days) on fecal progestagen profiles that reflect reproductive activity (pregnancy and estrous cycles). In samples obtained from a pregnant female and stored for 30 days, there were no significant differences in fecal progestagen concentrations between the three treatment conditions. However, the mean concentrations of progestagens (± SE) in untreated samples increased significantly from 8.3 ± 0.3 µg/g wet weight feces at day 0 to 17.7 ± 5.1 µg/g feces at day 90, and 17.8 ± 4.7 µg/g feces at day 180. Samples that were collected from a pregnant female and stored in 80% MeOH or dried in the solar box correlated with controls (r=0.86 and 0.87, respectively; P<0.05) at day 180. In contrast, samples that were stored without preservatives for 180 days did not correlate with controls (r=0.35, P>0.05). Progestagen concentrations from samples of the estrous cycling female showed similar results. In conclusion, fecal samples dried in a solar box cooker or stored in 80% MeOH maintained absolute and relative progestagen concentrations for at least 180 days when they were stored outdoors and exposed to the climatic conditions of central Florida. Both methods can have significant applications for the study of reproductive events in areas where access to electricity is limited. Zoo Biol 23:291–300, 2004. © 2004 Wiley-Liss, Inc.