Use of group-specific antibodies to detect fecal progesterone metabolites during the estrous cycle of cows |
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| Institution: | 1. Institute of Biochemistry & Ludwig Boltzmann Institute of Veterinary Endocrinology University of Veterinary Medicine, Josef Baumann-Gasse 1, A-1210 Vienna, Austria;2. Clinic of Obstetrics, Gynecology and Andrology University of Veterinary Medicine, Josef Baumann-Gasse 1, A-1210 Vienna, Austria;1. Research Station of the Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Ruciane-Nida, Poland;2. Department of Immunology and Pathology of Reproduction, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland;3. Department and Clinic of Reproduction, Faculty of Veterinary Medicine, Wroc?aw University of Environmental and Life Science, Wroc?aw, Poland;4. Department of Animal Reproduction, University of Warmia and Mazury, Olsztyn, Poland;5. Department of Pathology, Faculty of Veterinary Medicine, Wroc?aw University of Environmental and Life Science, Wroc?aw, Poland;1. College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China;2. College of Forestry, Northwest A&F University, Yangling, Shaanxi, 712100, China;3. Biology Research Centre of Qin Mountains Wildlife, Northwest A&F University, Yangling, Shaanxi, 712100, China;4. College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China;5. Shaanxi Reed Musk Deer Industry Co., Ltd., Meixian, Shaanxi, 722307, China;1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China;2. Department of Special Animal Nutrition and Feed Science, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;1. Département de biologie, and Centre d’études Nordiques, Université Laval, Québec G1V 0A6, Canada;2. Reproductive Physiology, Toronto Zoo, 361A Old Finch Avenue, Toronto, Ontario, M1B 5K7, Canada;3. Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Alberta T2N 4Z6, Canada;4. Calgary Zoo, Alberta T2E 7V6, Canada;1. Department of Analytical Chemistry, University of Córdoba, Córdoba, Spain;2. CeiA3 Agroalimentary Excellence Campus, University of Córdoba, Córdoba, Spain;3. Maimónides Institute for Biomedical Research (IMIBIC)/University of Córdoba/Reina Sofía University Hospital, Córdoba, Spain;4. CIBER Fragilidad y Envejecimiento Saludable (CIBERfes), Instituto de Salud Carlos III, Spain;5. Department of Genetic, University of Córdoba, Córdoba, Spain |
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| Abstract: | Progesterone is metabolized to pregnanediones and hydroxylated pregnanes prior to its fecal excretion. Therefore, use of progesterone antibodies underestimates the actual amount of fecal metabolites. To improve the methodology of noninvasive fecal progesterone metabolite analysis, enzymeimmunoassays (EIA) using group-specific antibodies against 5-reduced 20-oxo-pregnane-C3-conjugates were developed. Fecal and milk samples were collected at 1- to 2-d intervals during the morning and evening milking throughout 1 estrous cycle in dairy cows (n = 12). Six immunoreactive metabolites were detected in the feces with high performance liquid chromatography (HPLC), eluting as 5α- and 5β-reduced pregnanes containing a 20-oxo-group (20-oxo-pregnanes). Fecal samples of 3 cows were analyzed by 3 EIAs using antibodies against 4-pregnene-6α-ol-3,20-dione 6HS:BSA (6HS-progesterone), 5α-pregnane-3β-ol-20-one 3HS:BSA and 5β-pregnane-3β-ol-20-one 3HS:BSA, respectively. The follicular and luteal phases were identifiable with each EIA. Luteal phase values and the differences between mean follicular (Days 0 to 2 and 19 to 21) and luteal phase (Days 10 to 16) values obtained with the 5-pregnane EIAs were 3- to 4-fold higher than with the 6HS-progesterone EIA. Since results with the former 2 EIAs were almost identical, the remaining samples were only analyzed by the EIA for 5β-pregnane-3α-ol-20-one. Fecal 20-oxo-pregnane concentrations were parallel to milk progesterone values, but had a lag time of about 0.5 d; the coefficient of correlation (P < 0.001) was 0.76 (y = 155.2 × + 37.2). Fecal 20-oxo-pregnane concentrations during the follicular and luteal phase were 39.5 ± 2.2 and 341 ± 15.2 ng/g feces, respectively. In conclusion, fecal 20-oxo-pregnanes are significantly correlated to milk progesterone concentrations. They consist of several metabolites and compared to a 6HS-progesterone antibody, their evaluation was improved using antibodies against 5-reduced pregnanes. |
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