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A Non-Surgical Technique for the Collection of Uterine Fluid from the Mare¹

University of Florida, Gainesville 32611

Abstract

A non-surgical, transcervical technique was developed for collection of uterine secretions from unanesthetized horse mares. Uterine flushings were collected on days 4, 8, 12, 14, 16, 18 and 20 postovulation of the estrous cycle. The uterus was flushed with sterile .33 M saline, which was left in the uterus for a minimum of 5 minutes. During this time, the uterus was massaged per rectum to insure adequate mixing of luminal contents and saline. Fluid in the uterus was aspirated into sterile syringes and stored on ice until centrifuged. Centrifugation was carried out at 12,062 x g for 20 min in a refrigerated centrifuge. Flushings were filtered through a .45 µ: filter to remove bacteria and/or cellular debris. After the volume of each uterine flushing was measured, it was stored at –20 C until analyzed for protein. Volume of flushing medium collected from uteri (overall mean 106.3% fluid recovered) was not affected by day of the estrous cycle. Total amount of protein obtained in uterine flushings was affected (P<.025) by day of the estrous cycle with maximal amounts of recoverable protein in the mid-to-late luteal phase.

¹ Department of Animal Science, Florida Agricultural Experiment Station, Journal Series No. 332. This work was supported in part by N.I.H. Grant HD08560 and a grant from the Morris Animal Foundation.