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CHAPTER 4.2 subpopulation of superior sperm is cryopreserved. These sperm are more likely to withstand the cryopreservation protocol; additionally the harmful effect of seminal plasma is eliminated since almost all seminal plasma is discarded during the selection procedure and the subsequent washing of the pellet. It is likely that some poor freezing stallions might have experienced additional benefit if heterologous seminal plasma from a good freezing stallion is used instead of the homologous seminal plasma that was added back in the present study, this was however not the intention of the present study. Nevertheless it is clear that Androcoll-E can be beneficial to successfully cryopreserve stallion semen. Stallion selection however, plays a key role in the eventual outcome. 172 In conclusion, sperm selection using Androcoll-E prior to cryopreservation resulted in an overall increased post-thaw sperm quality. Stallions that produce ejaculates with low post-thaw PM might benefit from sperm selection prior to cryopreservation. As such only the best population of sperm is cryopreserved, which could result in good frozen semen. Androcoll-E can be beneficial to successfully cryopreserve stallion semen, but stallion freezability plays a role in the eventual outcome. Additional experiments need to be performed to evaluate the in vivo fertility of frozen thawed equine semen, selected with Androcoll-E prior to cryopreservation.
References CHAPTER 4.2 Barth A.D., Oko R.J. 1989. Abnormal morphology of bovine spermatozoa. Ames, Iowa: Iowa State University Press. Ecot P., Decuadro-Hansen G., Delhomme G., Vidament M. 2005. Evaluation of a cushioned centrifugation technique for processing equine semen for freezing. Animal Reproduction Science 89:245-8. Esteves S.C., Sharma R.K., Thomas Jr. A.J., Agarwal A. 2000 Improvement in motion characteristics and acrosome status in cryopreserved human spermatozoa by swim-up processing before freezing. Human Reproduction 15:2173-2179. Evenson D, Jost L. 2000. Sperm chromatin structure assay is useful for fertility assessment. Methods in Cell Science 22:169-189. Evenson D.P., Darzynkiewicz Z., Melamed M.R. 1980. Relation of mammalian sperm chromatin heterogeneity to fertility. Science 240:1131-1133. Garner D.L., Johnson L.A. 1995. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biology of Reproduction 53:276-84. Hansen C., Vermeiden T., Vermeiden J.P.W., Simmet C., Day B.C., Feitsma H. 2006 Comparison of FACSCount AF system, Improved Neubauer hemocytometer, Corning 254 photometer, SpermVision, UltiMate and NucleoCounter SP-100 for determination of sperm concentration of boar semen. Theriogenology 66:2188-2194. Henkel R.R., Schill W.-B. 2003. Sperm preparation for ART. Reproductive Biology and Endocrinology 1:108-130. Hoogewijs M., Rijsselaere T., De Vliegher S., Vanhaesebrouck E., De Schauwer C., Govaere J., Thys M., Hoflack G., Van Soom A., de Kruif A. 2010. Influence of different centrifugation protocols on equine semen preservation. Theriogenology 74:118-126. Januskauskas A., Johannisson A., Rodriguez-Martinez H. 2001. Assessment of sperm quality through fluorometry and sperm chromatin structure assay in relation to field fertility of frozenthawed semen from Swedish AI bulls. Theriogenology 55:947-961. Januskauskas A., Johannisson A., Rodriguez-Martinez H. 2003. Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure and field fertility. Theriogenology 60:743-758. Klinc P., Kosec M., Majdic G. 2005. Freezability of equine semen after glass beads column separation. Equine Veterinary Journal 37:43-47. Knop K., Hoffmann N., Rath D., Sieme H. 2005. Effects of cushioned centrifugation technique on sperm recovery and sperm quality in stallions with good and poor semen freezability. Animal Reproduction Science 89:294-7. 173
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AUTOMATED AND STANDARDIZED ANALYSIS
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AUTOMATED AND STANDARDIZED ANALYSIS
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A Area AI Artificial Insemination A
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PREFACE The first recorded case of
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GENERAL INTRODUCTION CHAPTER 1 3
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1.1.1. Introduction CHAPTER 1.1 The
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1.1.4. Concentration CHAPTER 1.1 Ac
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Fluorescent Activated Cell Sorter C
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CHAPTER 1.1 preparation (D, µm) is
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CHAPTER 1.1 These chambers are freq
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CHAPTER 1.1 it is not possible to o
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CHAPTER 1.1 overview of common abno
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CHAPTER 1.1 head, (B5) pyriform hea
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CHAPTER 1.1 The acrosome can be eva
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CHAPTER 1.1 The sperm chromatin str
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Collection and processing of equine
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CHAPTER 1.2 30 (a) Colorado State U
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CHAPTER 1.2 can be induced pharmaco
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CHAPTER 1.2 centrifugation time and
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CHAPTER 1.2 36 → Colloid centrifu
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CHAPTER 1.2 of a stallion stored fo
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CHAPTER 1.2 1.2.4. Cryopreservation
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CHAPTER 1.2 0.5 mL straws, and glyc
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CHAPTER 1.2 determined if the final
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CHAPTER 1.2 46 Chemically defined e
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CHAPTER 1.2 48 Cryopreservation - t
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1.3.1. Standards for equine AI dose
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CHAPTER 1.3 The use of density grad
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CHAPTER 1 Avery B., Greve T. 1995.
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CHAPTER 1 Cheng F.-P., Fazeli A., V
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CHAPTER 1 Flipse R.J., Patton S., A
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CHAPTER 1 Kuisma P., Andersson M.,
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CHAPTER 1 Medeiros A.S.L., Gomes G.
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CHAPTER 1 Pickett B.W. 1993. Collec
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CHAPTER 1 Taberner E., Morató R.,
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CHAPTER 1 Waite J.A., Love C.C., Br
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CHAPTER 2 As evidenced in the gener
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3.1 Influence of technical settings
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CHAPTER 3.1 a CEROS (Hamilton-Thorn
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CHAPTER 3.1 Progressive Motility CA
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CHAPTER 3.1 than at least they shou
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3.2.1. Abstract CHAPTER 3.2 Sperm m
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CHAPTER 3.2 depth but not the chamb
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3.2.3.3. The different chambers and
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CHAPTER 3.2 Finally, Pearson coeffi
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90 80 70 60 50 40 30 20 10 0 drop f
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ISAS20rM Makler10r WHO 120 120 120
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60 PM Rapid 50 40 30 20 10 0 CHAPTE
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Table 5. Averages (± standard erro
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CHAPTER 3.2 sperm (tail) which coul
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CHAPTER 3.2 Hoogewijs M.K., Govaere
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CHAPTER 3.2 Spizziri B.E., Fox M.H.
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3.3.1. Abstract CHAPTER 3.3 Routine
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CHAPTER 3.3 morphological abnormali
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CHAPTER 3.3 The extended semen was
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CHAPTER 3.3 Table 1: Average values
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Repeatability Agreement SQA-Ve PM (
Page 129: References CHAPTER 3.3 Arunvipas P.
Page 133 and 134: 3.4.1. Abstract CHAPTER 3.4 In this
Page 135 and 136: CHAPTER 3.4 therefore in these expe
Page 137 and 138: CHAPTER 3.4 The averages obtained a
Page 139 and 140: 3.4.4.2. Experiment 2 CHAPTER 3.4 F
Page 141: References CHAPTER 3.4 Arunvipas P.
Page 145: 4.1 Influence of different centrifu
Page 148 and 149: CHAPTER 4.1 4.1.2. Introduction 140
Page 150 and 151: CHAPTER 4.1 142 4.1.3.3. Semen proc
Page 152 and 153: CHAPTER 4.1 144 In the experiment 3
Page 154 and 155: CHAPTER 4.1 centrifugation (T1) and
Page 156 and 157: CHAPTER 4.1 (non CP vs CP) was sign
Page 158 and 159: 35 B 80 A 33 beat cross frequency (
Page 160 and 161: CHAPTER 4.1 152 4.1.4.2. EXPERIMENT
Page 162 and 163: CHAPTER 4.1 Love and coworkers (200
Page 164 and 165: CHAPTER 4.1 Love C.C., Brinsko S.P.
Page 167 and 168: 4.2.1. Abstract CHAPTER 4.2 The inc
Page 169 and 170: 4.2.3. Materials and methods 4.2.3.
Page 171 and 172: CHAPTER 4.2 For the SLC group, 15 m
Page 173 and 174: CHAPTER 4.2 adding 600 µL of AO st
Page 175 and 176: 4.2.3. Results 4.2.3.1. Sperm quali
Page 177 and 178: CHAPTER 4.2 The aforementioned “T
Page 179: CHAPTER 4.2 Sperm yields markedly d
Page 183: CHAPTER 4.2 Waite J.A., Love C.C.,
Page 187 and 188: CHAPTER 5 This thesis originated fr
Page 189 and 190: CHAPTER 5.1 Fig. 1. Schematic prese
Page 191 and 192: CHAPTER 5.1 higher than the current
Page 193 and 194: CHAPTER 5.1 al., 2007) showing a cl
Page 195 and 196: CHAPTER 5.1 treatment strategy for
Page 197 and 198: CHAPTER 5.2 supernatant allowing sp
Page 199 and 200: CHAPTER 5.2 such keeping the pellet
Page 201 and 202: CHAPTER 5.2 components of the semin
Page 203 and 204: Final reflections 5.4 Actual change
Page 205 and 206: CHAPTER 5 Dillon R.H., Fauci L.J.,
Page 207: CHAPTER 5 Suárez S.S., Osman R.A.
Page 210 and 211: SUMMARY technique that enables proc
Page 212 and 213: SUMMARY from stallions previously c
Page 214 and 215: SAMENVATTING beweeglijkheid (Hoofds
Page 216 and 217: SAMENVATTING de spermacellen gesele
Page 219 and 220: DANKWOORD Het obligatoire dankwoord
Page 221: DANKWOORD dankzij jou en ze zijn er
Page 225: CURRICULUM VITAE Maarten Hoogewijs
Page 228 and 229: BIBLIOGRAPHY Govaere J., Ducatelle
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BIBLIOGRAPHY ABSTRACTS AND PROCEEDI
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FUNDAMENTAL ASPECTS OF CRYOPRESERVA
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ADDENDUM I presence of these CPAs a
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ADDENDUM I Fig. 3. Theoretical curv
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ADDENDUM II DETAILED LOADING TECHNI
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In m’n hoofd is alles heel eenvou
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