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Since the successful establishment of a fistula of the vas deferens in the ram by White, Larson & Wales (1959), various techniques have been devised for the collection of epididymal spermatozoa from bulls, rams and boars. Wierzbowski & Wierzchos (1969) list authors who have undertaken such work. Recently, we have developed a fistula method for the regular collection ofepididymal spermatozoa from the rat. Because of its extremely small diameter, the rat's vas deferens is unsuitable for cannulation and when brought directly to the surface of the skin, it soon becomes obstructed and covered by closure of the skin above it. It can, however, be anastomosed either to the small or to the large intestine by simple implantation and will generally remain open and continue to function indefinitely. Utilizing this principle to construct a fistula of the vas deferens, the following procedure has now been frequently undertaken in mature Sprague-Dawley rats (Charles River CD strain). Under Nembutal anaesthesia, the abdomen was opened through a lower midline incision. The end of the caecum was carefully isolated from the caecum proper so that a good blood supply was preserved. A longitudinal opening, about 1 cm in length, was then made in the antimesenteric border of this isolated segment thus forming a pouch. In each dorsolateral aspect of the pouch, a small stab wound was made through which the vasa deferentia were brought out by means of traction sutures. The vasa deferentia were then divided and secured in position by a single fine suture. The four quadrants of the pouch were sutured to the parietal peritoneum to prevent herniation and the wound was closed about the pouch orifice. A small semicircle of skin was excised on either side of the pouch orifice to assist eversión of the stoma and the latter was then sutured to the surrounding skin, leaving an opening approximately 1 cm in diameter. After the operation, the pouch was occasionally flushed with a small quantity of saline until healing had been established. Text-figure 1 illustrates the various steps undertaken during the construction of the fistula. Within a few days of the operation, approximately 0-005 ml of spermatozoa

A TECHNIQUE FOR THE COLLECTION BY FISTULA OF EPIDIDYMAL SPERMATOZOA FROM THE RAT