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ABSTRACT TURNER, FRANCES BYRD. Evaluating Relationships ABSTRACT TURNER, FRANCES BYRD. Evaluating Relationships Between Various Estimates of Semen Quality Based on Fertility Assessments of Boars (Under the Direction of Dr. William L. Flowers). In commercial swine production, boars are an integral part of profitability. Since 99% of breeding in the swine industry uses artificial insemination, a boar can breed between 10,000 and 15,000 sows over his lifetime. Current measurements of semen quality used in the industry have not been studied in terms of their association with boar fertility. The objective of this study was to determine whether and to what degree common assessments of semen quality in boars are related to their fertility. Boars (n = 12) that were between two and two and a half years of age were used. Heterospermic insemations and paternity testing of the piglets were used to group these boars into High, Medium, and Low fertility groups. Average number of pigs sired per litter by boars in the High (n=3), Medium (n=6), and Low (n=3) groups were 71.2 + 0.7%, 51.2 + 1.5%, and 25.5 + 4.6%, respectively. The assumption was made that there was a positive relationship between the proportion of piglets sired in a litter and boar fertility. Both analysis of variance and multiple regression procedures were used to determine relationships between fertility and 76 different estimates of semen quality. Thirteen dealt with various anatomical and functional aspects of spermatozoa, while 63 were associated with proteins in seminal plasma. Proportions of motile spermatozoa (p < 0.075); spermatozoa exhibiting progressive forward motility (p < 0.075); spermatozoa with normal acrosome morphology (p < 0.05); and spermatozoa undergoing an acrosome reaction (p < 0.05) and the average straight line distance traveled by motile spermatozoa (p < 0.05) exhibited a positive relationship with fertility. Boars in the High group were higher than their counterparts in the Low group and boars in the Medium group being intermediate between the two. The average straight line velocity of motile spermatozoa was higher in the High and Medium boars compared with the Low boars. Concentrations of three known and 60 unknown seminal plasma proteins were examined in ejaculates from High, Medium, and Low fertility boars. Of the three known proteins, HSP-70, TNF-α, and TGF-β, only HSP-70 had a positive, linear association (p < 0.05) with boar fertility. Of the 60 seminal plasma proteins that were identified via two dimensional gel electrophoresis, only 10 had either positive or negative relationships with boar fertility. Two – a 26 kD/7.2 pI protein and a 28 kD/6.6 pI protein – exhibited a positive linear (p < 0.05) with fertility; and three – a 19 kD/9.2 pI protein, a 35 kD/6.3 pI protein, and an 186 kD/8.7 pI protein – had negative associations (p < 0.05) with fertility. For the remaining five proteins, 46 kD/6.8 pI (p < 0.10) and 59 kD/5.2 pI proteins were greater in High boars compared with Medium and Low boars. In contrast, concentrations of 19 kD/8.5 pI, 20 kD/6.2 pI, 40 kD/5.0 pI, and 111 kD/8.7 pI proteins were higher (p < 0.075) in Low than Medium and High boars. Results from the multiple regression analyses showed that three variables accounted for 87.6% of the variation in the average number of pigs sired per litter. These were concentrations of the 28 kD/6.6 pI protein (partial R2 = 0.6619; p < 0.0001); proportion of acrosome-reacted spermatozoa (partial R2 = 0.1549; p = 0.0002); and concentrations of the 17 kD/3.8 pI protein (partial R2 = 0.0590; p = 0.0090). In conclusion, two seminal plasma proteins and the proportion of spermatozoa that are acrosome-reacted per ejaculate hold the most promise for assessing boar fertility. Evaluating Relationships between Various Estimates of Semen Quality Based on Fertility Assessments of Boars by Frances Byrd Turner A thesis submitted to the Graduate Faculty of North Carolina State University In partial fulfillment of the Requirements for the degree of Master of Science Animal Science Raleigh, North Carolina May 15, 2008 APPROVED BY: _______________________________ ______________________________ Dr. William L. Flowers Dr. Scott Whisnant Committee Chair ________________________________ Dr. Glen W. Almond BIOGRAPHY Frances Turner was born on February 4th, 1984 in Charlotte, North Carolina to Tim and Sarah Turner. She has two older sisters Virginia and Rachel and one younger sister Lila. She attended Charlotte Latin School in Charlotte, North Carolina for kindergarten through high school. She graduated from Charlotte Latin School in 2002. After graduation from high school Frances came to Raleigh to attend North Carolina State University. She Graduated with her Bachelors of Science in Animal Science in May 2006. In August of 2006 she began to pursue her Masters of Science in Animal Science at North Carolina State University. ii ACKNOWLEDGEMENTS I would like to extend a special thank you to Dr. William Flowers. He has been a wonderful mentor through my undergraduate work and my graduate work. His knowledge, guidance, and encouragement have helped me with my success in this graduate program and other aspects of life. I would also like to say thank you to my graduate committee members Dr. Scott Whisnant and Dr. Glen Almond for their support and guidance. I would like to say a special thank you to Kara Stewart and Marcia Seal for all of their lab expertise and support. I would also like to say thank you to Lisa Thompson, Christy Kinney and Sara Shute for their help and friendship. I also would like to thank all of the graduate students for being supportive and helpful through out my stay as a graduate student. An exceptional thank you goes out to my family. To my Mom and Dad without all of your encouragement and support of my every endeavor I could not have gotten this far. I would also like to send a big thank you out to my sisters for all their encouragement and love. iii TABLE OF CONTENTS LIST OF TABLES ................................................................................................................ vii LIST OF FIGURES .................................................................................................................ix EVALUATING RELATIONSHIPS BETWEEN VARIOUS ESTIMATES OF SEMEN QUALITY BASED ON FERTILITY ASSESMENTS OF BOARS .................................... 1 LITERATURE REVIEW ........................................................................................................ 2 Introduction ................................................................................................................. 2 Oocyte ........................................................................................................................... 3 Sperm Production......................................................................................................... 9 Sperm Transport; Sperm Capacitation; and the Acrosome Reaction ..................... 14 Techniques used to Estimate Semen Quality and Boar Fertility ............................ 27 Visual Estimates ............................................................................................ 27 Semen Volume and Concentration of Spermatozoa .................................... 28 Normal Morphology ...................................................................................... 28 Motility of Spermatozoa ................................................................................ 29 Acrosome and Plasma Membrane Integrity ................................................ 30 Flow Cytometry ............................................................................................. 30 Acrosin Activity of Spermatozoa................................................................... 31 Sperm Binding Tests ...................................................................................... 32 Motility and Fertility ..................................................................................... 34 Sperm Chromatin Structure .......................................................................... 34 Measurements of Non-Sperm Components in Seminal Plasma.................. 36 iv Seminal Plasma Proteins .............................................................................. 37 WORKS CITED ..................................................................................................................... 41 INTRODUCTION .................................................................................................................. 49 MATERIALS AND METHODS .......................................................................................... 49 Experimental Design.................................................................................................. 49 Experimental Animals ............................................................................................... 50 Semen Collection ....................................................................................................... 52 Semen Preperation and Insemination for Heterospermic Matings ......................... 53 Paternity Testing ........................................................................................................ 54 Semen Quality Evaluations ....................................................................................... 56 Motility and Mobitlity.................................................................................... 56 Proportion of Morphologically Normal Acrosomes ................................... 57 Proportion of Spermatozoa with Acrosin
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