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.

Abstract

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...

Early Pregnancy Diagnosis in the Sow by Fecal Gestagen Measurement Using a Bovine Milk Progesterone Qualitative Test EIA Kit.

“…Feces was collected and preserved according to a method described in a previous report [18]. Fecal collection was carried out while each sow remained unrestrained in her stall.…”

“…Because the previous study [18] showed that the criterion for positive pregnancy diagnosis was a gestagen concentration of more than 200 ng/g of fecal weight or 5 ng/ml of fecal solution [18], a standard fecal solution was prepared with at a progesterone concentration of 5 ng/ml. The method of preparation was the same as that used in the previous study [18].…”

“…The method of preparation was the same as that used in the previous study [18]. First, fecal samples were collected from 5 sows in estrus by a method described previously.…”

“…Measurements using the quantitative kit were carried out according to the method of the authors [18]. Table 1 shows the accuracy of pregnancy diagnosis determined by using qualitative measurements of the fecal solution gestagen concentration from day 21-25 of the last mating.…”

“…Various methods of early pregnancy diagnosis for sows have been reported: the rectal examination method [1][2][3][4], vaginal mucosal tissue examination tatively measure the gestagen concentration in feces in approximately 1.5 h and uses a measurement sample (i.e., feces) that is much easier to collect than blood is. This method is quick, requires no special equipment, and good results were obtained from the application of this method to early pregnancy diagnosis [18]. The present study consisted of an investigation of a bovine milk progesterone qualitative test EIA kit (qualitative kit) that can measure progesterone concentration in approximately 10 min, and whether or not it can be applied to the qualitative measurement of gestagen in the feces of sows and whether these results can be applied to early pregnancy diagnosis.…”