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Kinoshita K.,Kobe University | Inada S.,Kobe University | Seki K.,Kobe Municipal Oji Zoo | Sasaki A.,Tama Zoological Park | And 3 more authors.
PLoS ONE | Year: 2011

Knowledge of the basic reproductive physiology of snow leopards is required urgently in order to develop a suitable management conditions under captivity. In this study, the long-term monitoring of concentrations of three steroid hormones in fecal matter of three female snow leopards was performed using enzyme immunoassays: (1) estradiol-17β, (2) progesterone and (3) cortisol metabolite. Two of the female animals were housed with a male during the winter breeding season, and copulated around the day the estradiol-17β metabolite peaked subsequently becoming pregnant. The other female was treated in two different ways: (1) first housed with a male in all year round and then (2) in the winter season only. She did not mate with him on the first occasion, but did so latter around when estradiol-17β metabolite peaked, and became pseudopregnant. During pregnancy, progesterone metabolite concentrations increased for 92 or 94 days, with this period being approximately twice as long as in the pseudopregnant case (31, 42, 49 and 53 days). The levels of cortisol metabolite in the pseudopregnant female (1.35 μg/g) were significantly higher than in the pregnant females (0.33 and 0.24 μg/g) (P<0.05). Similarly, during the breeding season, the levels of estradiol-17β metabolite in the pseudopregnant female (2.18 μg/g) were significantly higher than those in the pregnant females (0.81 and 0.85 μg/g) (P<0.05). Unlike cortisol the average levels of estradiol-17β during the breeding season were independent of reproductive success. The hormone levels may also be related to housing conditions and the resulting reproductive success in female leopards. The female housed with a male during the non-breeding season had high levels of cortisol metabolites and low levels of estradiol-17β in the breeding season, and failed to become pregnant. This indicates that housing conditions in snow leopards may be an important factor for normal endocrine secretion and resulting breeding success. © 2011 Kinoshita et al. Source


Kinoshita K.,Kobe University | Morita H.,Kobe University | Miyazaki M.,Kobe University | Hama N.,Kobe Municipal Oji Zoo | And 6 more authors.
Analytical Methods | Year: 2010

The usefulness of near infrared spectroscopy (NIRS) to monitor urine estrogen concentrations was studied in order to determine optimal timing for breeding captive female giant pandas. NIR spectra of daily urine samples from a female giant panda (Ailuropoda melanoleuca) were acquired in the period between March 1st and 25th, 2007 (n = 53). Estrone-3-glucuronide (E1G) concentrations in the samples were also measured by enzyme immunoassay (EIA). Transmittance spectra of all urine samples were obtained in the wavelength range from 1100 to 2432 nm (excluding the range from 1884 to 2012 nm) with sample thickness of 1 mm. Partial least square regression was applied to the spectra and good correlation was obtained between E1G concentration measured by EIA and predicted values by NIR (R2 = 0.94, SECV = 10.04 ng ml-1). The results of both soft-independent modeling of class analogy (SIMCA) and moving principal component analysis (MPCA) could detect the time changes in E1G concentration as measured by EIA (the Pearson's correlation coefficients between E1G concentration and the interclass distances of SIMCA or the index of MPCA were r = 0.64 and r = 0.81 respectively, P < 0.01). As for MPCA index, the index sharply dropped on March 24th corresponding to the decrease of the E1G concentration indicating ovulation. Finally, artificial insemination was performed for 3 consecutive days including the peak day, March 24th, and the female became pregnant. These results indicated that NIRS and the following MPCA analysis of the respective urine spectral data could detect the changes of urinary hormones during estrous cycle at a nanogram level. The NIRS can find the optimal timing for breeding quicker and easier than EIA, so this technique can be useful for captive breeding of this threatened species. © 2010 The Royal Society of Chemistry. Source

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