Chpt8_ANS3319SemenEvalFreezeLab_10

Chpt8_ANS3319SemenEv - ANS 3319C Reproductive Physiology Endocrinology Lab Semen Evaluation Cryopreservation Bovine Equine Objectives 1 2 3 To

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ANS 3319C Reproductive Physiology & Endocrinology Lab Semen Evaluation & Cryopreservation: Bovine & Equine Objectives 1) To introduce the components of semen evaluation used in preparing semen for freezing (cryopreservation) using the beef bull as our model. 2) To provide “hands-on” experience in performing the steps of a semen evaluation. 3) To provide “hands-on” experience in the process of cryopreservation of equine semen. Evaluation of Bovine Semen for Cryopreservation Visual Examination of Semen Sample The gross appearance of freshly collected semen is usually the first measure of semen quality. Neat (unaltered) semen appears as a thick whitish to slightly yellowish fluid. The thickness of the semen sample is a reflection of the number of sperm present. There should be no odor associated with the semen sample. Potential odors are suggestive of an infection or the presence of urine, which could be detrimental to fertility of the sample. Other problems considered to be detrimental can also be detected in the color of the semen such as blood, urine, and feces, which cause the semen to be pink to brownish. White clumps or flakes indicate pus and the presence of an infection in the reproductive tract of the male. Sometimes debris from the semen collection site might also be found in the semen sample such as sand, dirt, straw or other bedding material. Ejaculates that are abnormal in color or appearance should be discarded at this point in the processing. Viability Components 1. Motility Motility of semen is one of the simplest viability characteristics to evaluate. Semen is diluted with an isotonic buffer and then examined either under a bright field microscope or a phase contrast microscope using a heated stage (35 - 39°C). Slides and coverslips are pre- warmed on a slide warmer prior to adding warm semen (35 - 39°C). Several fields of view are examined and the percentage of motile cells estimated to the nearest 5 or 10%. Although it is important to look for progressively motile sperm (sperm moving in a nearly straight line) it may be just as relevant in evaluating viability to determine if sperm are just motile at all (total motility, sperm being able to propel themselves with a beating tail). For the first time evaluators, the process of estimating percentages of motile sperm often appears difficult and inaccurate. Critical in this is the standardization of the procedures for diluting the semen and making the slides. During the observations on the microscope it is also important to use the fine focus and continually focus on sperm in different planes. Semen samples kept for processing should have greater than 60% motile sperm . Gross Motility- gross swirling pattern of undiluted semen looks like a large school of fish. Individual Motility
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This note was uploaded on 07/16/2011 for the course ANS 3319 C taught by Professor Yelich during the Spring '11 term at University of Florida.

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Chpt8_ANS3319SemenEv - ANS 3319C Reproductive Physiology Endocrinology Lab Semen Evaluation Cryopreservation Bovine Equine Objectives 1 2 3 To

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