This paper was presented at the 2011 National Wombat Conference

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Tamara Keeley The value of non-invasive hormone monitoring in captive and wild wombats for management and conservation This paper was presented at the 2011 National Wombat Conference National Wombat Conference Sponsored by The Wombat Protection Society

Tamara Keeley The Wildlife Reproductive Centre (WRC) at Taronga Western Plains Zoo specializes in the evaluation of reproductive and adrenal hormones in serum, saliva, urine and faeces in a variety of wildlife species. This information enhances our general knowledge about different species, and can be used to help inform management and conservation policies. Both experienced in wildlife endocrinology, Tamara Keeley has been a full-time employee of the WRC since 2005 and Kerry Fanson has fulfilled various contract positions between 2009-2011. The WRC collaborates with the University of Queensland and Queensland s Department of Environment and Resource Management to gain knowledge on the reproductive and adrenal physiology of the Southern and Northern hairy-nosed wombat for management and conservation purposes. The WRC has also provided non-invasive sample analysis for captive wombats from Australia Zoo and Dreamworld to provide information on the reproductive status of individual females for management purposes. The value of non-invasive hormone monitoring in captive and wild wombats for management and conservation Authors Tamara Keeley and Kerry Fanson Non-invasive hormone analysis is a useful tool for monitoring both captive and wild populations. Hormones are excreted in urine and faeces, so reproductive and adrenal/ stress physiology can be assessed without impacting the animal. Evaluating reproductive hormone concentrations can provide information of the reproductive status of an individual (cycling, pregnant) and increase our understanding of the reproductive biology of the species. Changes in adrenal hormone concentrations can be used to evaluate potential effects of management, transport, disease and environmental pressures on an individual or a population. In collaboration with other researchers and as a service for other zoos, our lab has used this technique to monitor reproductive and adrenal physiology in captive common and Southern hairy-nosed wombats. We have also monitored adrenal activity in naturally-occurring and reintroduced populations of Northern hairy-nosed wombats. There is vast potential to integrate this monitoring technique as a tool to monitor and improve in situ and ex situ management plans for all of the wombat species. Sponsors The Wombat Foundation Saving the Northern Hairy-nosed Wombat

The value of non-invasive hormone monitoring in captive and wild wombats for management and conservation Tamara Keeley and Kerry Fanson Wildlife Reproductive Centre Taronga Western Plains Zoo

Endocrine monitoring Endocrinology = Study of Hormones Most aspects of reproduction are mediated through hormones Basic information Understanding Basic Biology reproduction stress responses, adaptation behavior Evaluate among species differences

Applied uses Assess reproductive status Diagnose fertility problems Improve or control reproduction Evaluate seasonality Assess adrenal activity Monitor welfare and health

Marsupials Eastern, Western Grey Kangaroo Red Kangaroo Tasmanian Devil Tree Kangaroo Koala Possum Bilby Mahogany Glider Julia Creek Dunnart And...All 3 Wombat Species

Hormone Monitoring in Wombats Captive Bare Nosed Wombats (Service work) Evaluation of reproductive cycles Captive Southern Hairy-Nosed Wombats (UQ PhD Research) Evaluation of reproductive cycles Evaluation of adrenal hormones Stereotypies, enrichment, handling, environmental changes Wild Northern Hairy-Nosed Wombats (Qld DERM) Evaluation of adrenal hormones Adaptation

Non-Invasive Endocrine Monitoring Measurement of reproductive hormones and their metabolites in the feces or urine Non-invasive No impact on hormone expression Summary of endocrine activity Enables large scale, longitudinal studies

Methods of Monitoring Hormones Invasive - stressful Serum Urine Feces Non invasive - Difficult to collect Non-invasive easy to collect Occasional sampling Longitudinal Sampling Longitudinal sampling No time lag < 24 hours time lag Up to 48 hours time lag Snapshot not always representative of current status Original hormone of interest Protein and Steroid Hormones Index samples by creatinine Metabolized conjugates Steroid Hormones Pooled value Metabolites - more noise Steroid Hormones

Biological Validation of Methods Radio-labelled infusion Behavioural - Oestrus Physiological Event Pregnancy Stressor eg. trauma, disease Exogenous hormone challenge GnRH (FSH, LH, androgens) ACTH (corticosteroids)

Physiological Validation Adrenal Hormones Northern Hairy-Nosed Wombat 2000 1750 1500 1250 1000 750 500 250 0 21-Feb-14 28-Feb-14 07-Mar-14 14-Mar-14 21-Mar-14 28-Mar-14 04-Apr-14 11-Apr-14 18-Apr-14 25-Apr-14 02-May-14 09-May-14 16-May-14 23-May-14 30-May-14 06-Jun-14 Corticosteorne ng/g

Hormone Analysis

Follicular Development Ovulation CL Development LH Ovulation Oestrogen FSH Progesterone Luteal Phase Follicular Phase Luteal Phase

Faecal Hormone Analysis Progesterone Milly - Wombat Faecal Progesterone ng/g 600 500 400 300 200 Ovulation 100 0 03-Jan-14 10-Jan-14 17-Jan-14 24-Jan-14 31-Jan-14 07-Feb-14 14-Feb-14 21-Feb-14 Date Dreamworld

Urinary Hormone Analysis - Oestrogen Dozer - Wombat Mini Bus - Wombat Urinary Oestrogen metabolites ng/mg creatinine 80 60 40 20 0 Male Chasing Dozer Aggressive towards other females Urinary Oestrogen metabolites ng/mg creatinine 60 50 40 30 20 10 0 Date Date Australia Zoo

Benefits and Uses Ex Situ Captive Breeding Programs Confirm pregnancy Confirm cycling Adrenal Function Transport Captive conditions In Situ Seasonality Identification of cycling females Adrenal Function Adaptation Environmental pressures Disease

Acknowledgements We would like to thank. Collaborators for introducing us to the world of wombats Dreamworld Australia Zoo Qld Department of Environment and Resource Management UQ