Development and Evaluation of a Rapid Enzyme-immunoassay System for Measurement of the Urinary Concentration of Estrone-3-glucuronide in a Female Giant Panda (Ailuropoda melanoleuca)

Abstract

Abstract. To detect estrus for reproductive management, and to determine the relationship between urinary estrogen and estrous behavior, in a female giant panda, we developed and evaluated a rapid enzyme immunoassay (EIA) system for urinary Estrone-3-glucuronide (E1G) using commercial reagents. The developed EIA system took only around 3 hours, including all procedures to obtain a result. It indicated good reproducibility (intra-assay CV of 5.16%, interassay CV of 15.4%) and sensitivity (lowest standard concentration was 0.0156 ng/ml) for measurement of the urinary concentrations of E1G in the giant panda. There was a positive correlation (r=0.934) with the data for estrone (E1) in the same samples, as measured by radioimmunoassay (RIA) performed in a commercial laboratory. The changes in the E1G concentrations were almost synchronous with the changes in E1 assayed by RIA in urine collected during 4 consecutive estrous seasons. The dynamics of urinary E1G measured by this system highly correlated with the occurrence of the presenting estrous behavior in the giant panda. The above results indicate that this assay system may be normally, rapidly and practically used for measurement of the urinary concentration of E1G in the giant panda. Key words: Giant panda (Ailuropoda melanoleuca), Enzyme immunoassay, Estrone-3-glucuronide, Estrous behavior (J. Reprod. Dev. 54: [281][282][283][284][285] 2008) he giant panda (Ailuropoda melanoleuca) is a critically endangered species. Its natural habitat is located and limited to Western China at elevations of 2,000 m to 4,000 m. The actual population is estimated to be no more than 1,500 individuals [1]. The giant panda is categorized as "Endangered" on the Red List of Threatened Species published by the International Union for Conservation of Nature and Natural Resources [2].Female giant pandas have a single annual estrus period and generally are in estrus for only a few days between February and May [3]. This characteristic constitutes a major limitation on production of new born in giant pandas.At the Kobe Municipal Oji Zoo, a collaborative project concerning reproduction of the giant panda has been conducted with China since 2000. Since the main purpose of this project is production of baby pandas, the primary work required is establishment of a rapid assay system to measure the sex steroid hormones to evaluate the giant panda's reproductive status and especially to detect estrus by measuring estrogen concentrations in the female for natural and artificial breeding.Because zoo and wild animals are not domesticated or are untrained, frequent sampling of blood is very difficult, and so urine or feces, which are easily collected without being invasive to the animals, are used to measure hormones. In Japan, another study of reproduction of the giant panda has been underway at Ueno Zoological Gardens, Tokyo, since 1972. They have succeeded in production of giant panda by artificial insemination (AI) [4]; however, they have also experienced unsuccessful reproduction in a fema...

Urinary sex steroid hormone and placental leucine aminopeptidase concentration differences between live births and stillbirth of Bornean orangutans (Pongo pygmaeus)

“…Urine supernatant was used for the assays. Urine samples were Urine concentrations of E 1 G and PdG were assayed using an enzyme immunoassay using the same methods and reagents as described by Hama et al 22,23 The polyclonal E 1 G antiserum (FKA-224E;…”

Urinary estrogens, progesterone, and LH changes during normal menstrual cycles of a captive female pileated gibbon (Hylobates pileatus)

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Development of an Enzyme Immunoassay for Urinary Pregnanediol-3-Glucuronide in a Female Giant Panda (Ailuropoda melanoleuca)

“…The concentrations of standards used in this study were between 0.03125 and 2.0 ng/well. The EIA buffer, coating buffer, wash buffer, substrate buffer and 2nd antibody used in the present study were the same as those used in a previous report [4]. A plate was coated with 2nd antibody and washed with wash buffer 2 times.…”