Pregnancy diagnosis in zoo animals by estrogen determination in feces

Safar‐Hermann, Nina; Ismail, Mohammed; Choi, H. S.; Möstl, Erich; Bamberg, E.

doi:10.1002/zoo.1430060209

Cited by 35 publications

(13 citation statements)

References 9 publications

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“…In recent years, the measurement of the steroid hormone concentration in feces has been reported to be a useful diagnostic tool for wild animals [8,10,12,19,21,26] and pigs [1,2,7,13,20,25], blood collection from which is especially difficult.…”

Section: Extraction Of E1s: E1s Was Extracted From Fecal Solutionmentioning

confidence: 99%

Fecal Estrone Sulfate Profile in Sows during Gestation.

Ohtaki

Moriyoshi

Nakada

et al. 1999

The Journal of Veterinary Medical Science

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ABSTRACT. The aim of this study was to establish radioimmunoassay (RIA) for fecal estrone sulfate (E1S) and to elucidate changes in fecal E1S during pregnancy in the sow. Fecal E1S was extracted on a commercially available solid phase column, and the E1S fraction obtained was subjected to RIA. The sensitivity of the RIA was 8.5 pg/tube. The intra-and inter-assay coefficients of variation were 8.8-8.9% and 10.7-14.2%, respectively. Mean recovery for E1S added to fecal samples was as high as 95.0%. A significant positive correlation (r=0.904, n=147 p<0.0001) existed between fecal and plasma E1S concentrations. Mean E1S concentration in feces and plasma fluctuated exhibiting two peaks. The first peak of E1S concentration was evident on days 28-32 in feces and on days 26-30 in plasma. The E1S concentration in both feces and plasma remained at baseline levels during mid-pregnancy, but began to rise gradually around days 72-82 and 70-80, in feces and plasma respectively, and reached a peak concentration on days 110-114. Following parturition, the concentration of E1S in plasma declined rapidly, but there was a two-day delay before a decline in fecal E1S. Apart from this two-day delay, changes in fecal E1S were similar to those in plasma E1S. The study indicates that the measurement of E1S in feces could be a useful tool for early pregnancy diagnosis and for monitoring fetal development in sows and gilts.-KEY WORDS: estrone sulfate, feces, pregnancy, radioimmunoassay, sow.J. Vet. Med. Sci. 61(6): 661-665, 1999 our department were used for the present study. The sows were artificially inseminated on the first day of estrus. Fecal samples were collected from each sow at intervals of two days during pregnancy, as described previously by Moriyoshi et al. [13]. Briefly, a plastic glove for rectal examination was worn on the hand which was inserted into the rectum of sows. An appropriate amount of impacted feces was taken from the rectum and immediately aliquoted in 2 g amounts. The aliquots were put into plastic test tubes each containing 8 ml of 0.01 M phosphate buffer with 0.1% of bovine serum albumin (BSA; albumin, bovine fraction V, Sigma-Chemicals Co., St. Louis, U.S.A), and then taken back to the laboratory. After thoroughly shaking on a Direct Mix TS-50 (Thermal Kagaku Sangyo Co., Ltd., Japan), the solution was centrifuged at 1,700 × g for 15 min, and the supernatant was frozen at 20°C as the fecal solution until assayed for E1S. Immediately after fecal collection, blood samples were also collected from the median tail vein into heparinized test tubes. Plasma was separated by immediate centrifugation (1,700 × g, 15 min) and kept frozen at 20°C until assayed for E1S. In order to use as E1S standard for fecal samples, E1S-free feces were collected from 5 castrated boars. The E1S-free fecal solutions prepared as described above were pooled. Preparation of standard fecal E1S solution: Fecal samples were collected from 5 castrated boars and the fecal solutions prepared were pooled as described above. These were used fo...

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Section: Extraction Of E1s: E1s Was Extracted From Fecal Solutionmentioning

confidence: 99%

Fecal Estrone Sulfate Profile in Sows during Gestation.

Ohtaki

Moriyoshi

Nakada

et al. 1999

The Journal of Veterinary Medical Science

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“…Fecal samples are easier to collect than urine samples and the current trend in noninvasive monitoring of reproductive function suggests their increasing importance [Lasley and Kirkpatrick, 19911. Several assays based on fecal sample analysis have been shown to be reliable markers of luteal activity andior pregnancy in domestic and nondomestic species [Mostl et al, 1983;Bamberg et al, 1984Bamberg et al, , 1991Choi et al, 1987;Safar-Hermann et al, 1987;Desaulniers et al, 1989;Messier et al, 1990;Kirkpatrick et al, 1991;Lucas et al, 1991;Wasser et al, 1991;Schwarzenberger et al, 1991Schwarzenberger et al, , 1992. The purpose of the present study was to establish a method for fecal progestagen analysis during the estrous cycle and pregnancy in the okapi.…”

Section: Introductionmentioning

confidence: 99%

Fecal progestagen evaluations to monitor the estrous cycle and pregnancy in the okapi (Okapia johnstoni)

et al. 1993

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The present study was conducted to establish a noninvasive method of reproductive monitoring in the okapi (Okapia johnstmi). Fecal samples were collected three times a week from nonpregnant okapis (n = 3) for periods of 2, 9, and 23 months, respectively, and for 2 months each from pregnant okapis (n = 4) at different stages of gestation. Samples were analyzed with an enzyme-immunoassay (EIA), using an antibody against pregnanediol, and the results are considered as measurements of unconjugated total immunoreactive progestagens (Pd-Pgs). Mean values of Pd-Pgs during the follicular (FP) and the luteal phases (LP) of the estrous cycles were 0.6 * 0.1 Kg/g and 6.1 * 0.3 Kg/g feces, respectively.Matings occurred at the terminations of the LP. By dividing the number of entire LPs into the time over which samples were available, average estrous cycle lengths in two okapis were estimated to be 15.5 (n = 11 LP) and 15.8 (n = 36 LP) days, respectively. In three animals, gestation lengths of 423, 424, and 431 days were calculated by observed matings. During days -280 to -220 before parturition, the fecal Pd-Pgs constantly increased from about 20-60 pg/g feces. Values were 100-350 pg/g during the last third of gestation. Values decreased in the week before parturition, and a continuous decline to FP values was observed within 3-4 days postpartum. During the 2 months postpartum investigated in one animal, the Pd-Pgs were in the FP range except one LP 3 weeks postpartum. It was concluded that fecal Pd-Pgs in female okapis are present in a ratio of -1 : lo:> 100 during FP, LP, and late pregnancy, respectively. Their measurement by EIA enables noninvasive monitoring of the estrous cycle and pregnancy diagnosis.

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“…These have been used as measurement samples to measure the steroid hormone concentration in animals, and good results have been reported in many cases. The measurement of the steroid hormone concentration in feces has been reported to be a useful method for wild animals and pigs, from which blood collection is especially difficult [2,4,14,22,25,37,39]. However, most of these reports on the use of fecal samples used radioimmunoassay (RIA) for the measurements, and as RIA can only be performed at a very limited number of laboratories due to various restrictions, and requires a long time for measurement, these reported methods of measurement using fecal samples have low applicability to the clinical field of veterinary medicine.…”

Section: Standard Fecal Gestagen Solution Preparation: Fecalmentioning

confidence: 99%

Measurement of Gestagen Concentration in Feces Using a Bovine Milk Progesterone Quantitative Test EIA Kit and Its Application to Early Pregnancy Diagnosis in the Sow.

Moriyoshi

Nozoki

Ohtaki

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. We attempted to measure the gestagen concentration in the feces of pigs by using a commercial bovine milk progesterone quantitative test EIA kit, and investigated the possibility of applying of this method of gestagen concentration measurement to early pregnancy diagnosis in the sow. Feces were collected from the rectum of the pig, and 0.5 g of the feces was placed in 20 ml of distilled water, stirred, and centrifuged. The supernatant was used as the fecal solution for measurement of gestagen. The procedure used for measuring gestagen in feces was the same as that for the measurement of progesterone in milk, except that a standard fecal gestagen solution (0.5-30.0 ng/ml) was prepared by the authors in the laboratory. The sensitivity of measurement using this method was 0.80 ng/ ml, or 32.0 ng/g of fecal weight. The recovery was 105.2-105.6%. Intra-assay coeffecients of variation (CVs) were 2.8-8.5%. The interassay CVs were 7.4-10.2%. Gestagen concentrations in feces measured by the present method and progesterone concentrations in peripheral plasma, collected at the same time as the feces, were highly correlated (r=0.98, p<0.001). The criteria for diagnosis of pregnancy based on the fecal gestagen level was positive for a gestagen level of 200 ng/g and negative for a gestagen level of < 200 ng/ g. When fecal gestagen measurements were applied to early pregnancy diagnosis in 149 sows, the accuracy of diagnosis from day 21 to day 25 after the last mating was 96.2% for positive cases (102/106) and 95.3% for negative cases (41/43). Thus, the results of this study show the quantitative measurement of the fecal gestagen concentration in the sow using a bovine milk quantitative test EIA kit is a practical method for early pregnancy diagnosis. -KEY WORDS: early pregnancy diagnosis, feces, gestagen, quantitative test EIA kit, sow.J. Vet. Med. Sci. 59(8): 695-701, 1997 easier to collect than blood. Measurements were carried out using a commercial bovine milk progesterone enzyme immunoassay (EIA) kit, which is quick and does not require any special equipment. The authors reported good results from the application of this method to early pregnancy diagnosis in sows [27][28][29]. In the present study, the same commercial bovine milk progesterone EIA kit was used for quantitative measurement of the gestagen concentration in samples of feces, which are even easier to collect than saliva, and the applicability of this method to early pregnancy diagnosis in the sow was investigated. MATERIALS AND METHODS Test kit:The kit used for gestagen quantitative measurement was an Ovucheck bovine milk progesterone EIA kit (Cambridge Veterinary Sciences Ltd., U.K.). The microtitre plate had 96 wells with sheep progesterone antibody pre-coated in each well. The reagents included enzyme-labeled antigen-binding solution (conjugate), substrate tablets, substrate tablet soluble buffer solution, and stopping solution, as well as four types of standard progesterone milk solutions (1.0, 5.0, 10.0, 30.0 ng/ml; standard milk solution was...

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Pregnancy diagnosis in zoo animals by estrogen determination in feces

Cited by 35 publications

References 9 publications

Fecal Estrone Sulfate Profile in Sows during Gestation.

Fecal Estrone Sulfate Profile in Sows during Gestation.

Fecal progestagen evaluations to monitor the estrous cycle and pregnancy in the okapi (Okapia johnstoni)

Measurement of Gestagen Concentration in Feces Using a Bovine Milk Progesterone Quantitative Test EIA Kit and Its Application to Early Pregnancy Diagnosis in the Sow.

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