Kirkpatrick, J. F., Kasman, L. H., Lasley,, B. L., & Turner, J. W. J. (1988). Pregnancy Determination in Uncaptured Feral Horses. J Wildl Manag, 52(2), 305–308.
Abstract: The urinary excretion of estrone sulfate ($\text{E}{1}\text{S}$) by 25 free-roaming feral horses (Equus caballus) was measured by radioimmunoassay applied to extracts of urine-soaked soil. Twelve of 15 mares having $\text{E}{1}\text{S}$ concentrations >1.0 mg/mg creatinine (x = 2.64 +- 1.02 [SD]) produced foals. All 10 mares with $\text{E}{1}\text{S}$ concentrations <1.0 mg/mg creatinine (x = 0.44 +- 0.26) did not foal. Extracting urine from soil and measuring $\text{E}{1}\text{S}$ and creatinine can be used to determine pregnancy in free-roaming feral horses without the stress of capture or immobilization.
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Kirkpatrick, J. F., Lasley, B. L., & Shideler, S. E. (1990). Urinary steroid evaluations to monitor ovarian function in exotic ungulates. Zoo Biol, 9(5), 341–348.
Abstract: A direct enzyme immunoassay (EIA) for non-specific urinary progesterone (Po) metabolites, utilizing a non-specific monoclonal antibody against pregnanediol-3-glucuronide, was evaluated for the purpose of assessing luteal function in equids. Urinary pregnanediol-3-glucuronide (PdG) and immunoreactive PdG-like conjugate (iPdG) concentrations, indexed by creatinine, were compared to plasma Po concentrations in non-conceptive ovarian cycles through two ovulations in four mares. High-performance liquid chromatography (HPLC) of urine from lutealphase mares and a pregnant zebra revealed an absence of significant concentrations of PdG and the presence of at least three immunoreactive compounds, all of which were more polar than PdG. The concentration of iPdG in the mare ranged from a nadir of approximately 3 ng/mg Cr at the time of ovulation to nearly 400 ng/mg Cr at the mid-luteal-phase peak and paralleled plasma Po concentrations. This non-radiometric assay for iPdG permits the assessment of ovulation, luteal formation and function, and luteolysis in unprocessed urine samples from domestic mares. Data from a single zebra indicate this approach also will permit simplified and non-invasive longitudinal studies of ovarian function among a wide range of Equidae.
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Kirkpatrick, J. F., Shldeler, S. E., Lasley, B. L., & Turner, J. W. J. (1991). Pregnancy determination in uncaptured feral horses by means of fecal steroid conjugates. Theriogenology, 35(4), 753–760.
Abstract: This study was carried out to develop an accurate, rapid and inexpensive method for diagnosing pregnancy in uncaptured feral horses by analysis of fecal steroid metabolites and to compare the accuracy of this method with diagnosis by urinary estrone conjugates (E(1)C). Paired urine and fecal samples were collected from 40 sexually mature feral mares during August and October. Urine samples were extracted directly from the soil and analyzed by enzymeimmunoassay (EIA) for E(1)C. Water extracts of fecal samples were assayed by EIA for E(1)C and nonspecific progesterone metabolites (iPdG). Urinary E(1)C, fecal E(1)C and fecal iPdG concentrations for seven mares which produced foals were 3.9 +/- 1.3 (SEM) mug/mg creatinine, 4.2 +/- 0.8 ng/g feces and 1.411 +/- 569.6 ng/g feces, respectively. Urinary E(1)C and fecal E(1)C and iPdG concentrations for the 33 mares which did not produce foals were 0.1 +/- 0.0 mug/mg creatinine and 0.5 +/- 0.1 and 32.8 +/- 4.5 ng/g feces, respectively. These differed (P < 0.01) from values in mares which produced foals.
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