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• W2922303743 abstract "Southern hairy-nosed wombats (SHNW - Lasiorhinus latifrons) do not breed well in captivity. To overcome this, a better understanding of their reproductive physiology is essential. While faecal hormone analysis has provided useful information regarding progesterone excretion in the SHNW, biologically relevant changes in oestrogens have proven difficult to detect, and protein hormones such as luteinizing hormone (LH) are not readily excreted in faeces. The analysis of urinary hormone metabolites is a valuable tool for assessing reproductive function in other wildlife species and has been linked to changes in reproductive behaviour, which can then be used as an effective non-invasive method of detecting oestrus and oestrous cycle characterisation for captive breeding programs. The overall aims of this research program were to: (1) determine if female captive SHNW could be conditioned for the reliable and frequent urine sample collection for this study, (2) develop and validate techniques to detect biologically relevant changes in urinary LH, oestrogens and progesterone metabolite concentrations in captive female SHNWs, (3) characterise the oestrous cycle of female SHNWs, (4) determine if changes in reproductive hormones could be mapped to changes in behaviours and urine sample characteristics, and (5) determine if male SHNWs could differentiate between oestrus and non-oestrus female urine. The overall objective of this research program was to increase our understanding of the female SHNW reproductive physiology and identify reliable, or improve current, methods of detecting oestrus to enhance captive breeding success of this species. Captive female SHNWs (n = 11) were classically conditioned to provide daily urine samples during the 2013 and 2014 breeding seasons. Initially, the animal was conditioned to a tactile stimulus (light tapping of the pericloacal region) and the presence of the collection tray under the rump during normal urination behaviour. Following three months of conditioning, collection success was 82%; however, some collection protocols were modified for timid or aggressive females, such as the use of a false floor, and aspiration of urine off a clean enclosure floor, to minimise stress associated with daily sample collection. Fresh urine samples were analysed for physical characteristics, including volume, specific gravity, pH, leukocytes and qualitative index of the number of urogenital epithelial cells. Samples were then stored at -20 oC for subsequent analysis of urinary creatinine, LH, oestradiol-17b (E2), estrone-3-glucuronide (E1C) and progesterone metabolites (P4M). Urinary reproductive hormone analysis techniques were validated using two exogenous hormone trials: (1) an injection of a high dose (4 and 10 g) of exogenous gonadotrophin releasing hormone (GnRH) to induce an LH surge that could be detected in urine; and (2) two sequential and increasing injections of equine chorionic gonadotrophin (150 and 300 IU) to stimulate the secretion of urinary E2, E1C and P4M . While serum and urinary LH increased significantly after the GnRH challenge, the collection of single daily urine samples was not sufficient for the detection of a naturally occurring pre-ovulatory LH surge. Similarly, biologically relevant changes in E1C concentrations were only detected after exogenous hormone injections and could not be used to accurately identify the different stages of naturally occurring oestrous cycles. While the analysis of urinary E2 resulted in higher overall oestrogen concentrations compared to E1C, E2 profiles were similar to E1C patterns, in that the different stages of the oestrous cycle were not reliably distinguishable. Further attempts to extract oestrogen metabolites from urine samples by enzyme hydrolysis or diethyl ether extraction also failed to improve the detection of distinct increases in oestrogen associated with oestrus. In contrast, the analysis for urinary P4M and changes in P4M concentrations could be used to identify the approximate onset and duration of both hormonally-induced and natural luteal phases. The mean oestrous cycle length (the beginning of one luteal phase to the beginning of the next) was 35.1 p 2.4 days, with a mean luteal phase of 20.8 p 1.3 days. There was a high level of between and within animal variation in oestrous cycle dynamics as cycles ranged from 23 to 47 days (luteal phases ranged from 12 to 33 days) with some females exhibiting both long and short oestrous cycles within a single breeding season. Changes in the physical characteristics of each urine sample were compared to changes in urinary P4M concentrations. When urinary P4M was l baseline values, urine volume decreased, whereas urine concentration (as evaluated by creatinine and specific gravity) and urogenital epithelial cells increased significantly. To determine if changes in urinary P4M could be mapped to changes in behaviour, the frequency and duration of 26 general, defensive and reproductive behaviours were analysed for seven females for eight days surrounding the onset of the luteal phase; three days when urinary P4M was l baseline values (D-3, D-2, D-1), three days when urinary P4M was g baseline values [D0 (classified as the first day where urinary P4M was above baseline for 3 consecutive days), D+1, D+2)] and two days which served as behaviour control days (D-14 and D+14) when reproductive behaviours were least likely to occur. Behavioural analysis showed that the duration of urination decreased significantly when P4M was l baseline values; however, there was no difference in the frequency or duration of feeding. Five of the seven females analysed exhibited pacing behaviour which increased significantly when P4M was l baseline values. Rump biting (defensive behaviour) by the female also increased when urinary P4M was l baseline values. Male-initiated urogenital sniffing was low when urinary P4M was l baseline values, and the lack of difference in olfactory behaviour of males exposed to oestrus and non-oestrus urine samples confirmed that males did not preferentially investigate oestrus samples and therefore were unlikely to be able identify receptive females by evaluating urine alone. The results from this research demonstrate that the collection and analysis of urine can be a valuable non-invasive tool for the reproductive assessment of captive female SHNW. The analysis of urinary P4M was effective for estimating the reproductive cycle dynamics of captive females; however, more research is required to refine techniques for the accurate assessment of biologically relevant changes in urinary LH and oestrogens. The collection of fresh urine samples from individual females and the analysis of urinary characteristics enabled the identification of markers which could be linked to changes in urination behaviour and locomotion, potentially useful non-invasive tools for monitoring the reproduction of this species and identify receptive females to optimise future breeding attempts." @default.
• W2922303743 created "2019-03-22" @default.
• W2922303743 creator A5009866306 @default.
• W2922303743 date "2018-05-25" @default.
• W2922303743 modified "2023-09-24" @default.
• W2922303743 title "The use of urine as a tool to analyse the reproductive function of captive female southern hairy-nosed wombats (Lasiorhinus latifrons)" @default.
• W2922303743 doi "https://doi.org/10.14264/uql.2018.351" @default.
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