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CHAPTER 5.1 5.1.3. SQA-Ve in standardized semen analysis 186 The sperm quality analyzer (SQA) is not susceptible to similar operational variations as CASA since the device uses integrated algorithms that cannot be changed by the users, and only allow insertion of the tailor-made capillary. As such, the above mentioned issues of settings and counting chambers will not manipulate results, reducing potential sources of bias (Hoflack et al., 2005). Despite these advantages the present versions of the SQA-Ve should not be used for analyzing equine semen samples. So far, as shown in Chapter 3.3 and 3.4, the device is not precise enough and it lacks accuracy to report on equine semen quality. The improvements in the agreement of the SQA-Ve version 1.00.61 with CASA, as evidenced in Chapter 3.4, indicates that with further optimization of the algorithms the SQA might eventually be able to univocally report on equine semen quality. 5.1.4. Standardized semen analysis in relation to fertility So far, sperm motility only correlates poorly with stallion fertility (Jasko et al., 1992; Kuisma et al., 2006). Of course, a sperm cell needs more traits to fertilize, besides being motile. A spermatozoon should also be capable of initiating capacitation and the acrosome reaction, and should have intact DNA in order to be fertile. Mare factors interfere equally with early conception, and affect fertility chances as well. The introduction of actual standards in semen analysis might enable scientists all over the world to assess semen in the same way. As such more and more uniform data will come available which might lead to additional value of in vitro parameters in predicting in vivo fertility. Based on in vivo fertility data of the previous breeding season, a breeding stallion can be classified as being highly fertile, fertile or sub-fertile. For each of these categories, a different artificial insemination (AI) dose of progressive motile sperm might be required to maintain at least a comparable fertility level. Following every season, the fertility status should be recalculated and used during the following breeding season. The AI dose for stallions of unknown fertility (such as young, novice stallions) can be arbitrary set at the same dose as for fertile stallions. As soon as actual fertility data are available, true fertility should be calculated. Much can be debated about the fertility status of a stallion, but at the end fertility is determined by the interaction of stallion, mare and management factors (Fig. 4). Influence of stallion and mare on fertility is evident, but “management” factors such as cycle control, timing of AI,
CHAPTER 5.1 treatment strategy for endometritis, semen handling and so on will also interfere with the final fertility. In this thesis, the focus was partially on the quality of the semen and how it can be assessed (stallion factor influenced by management), and partially on preparation of AI doses (management factor influenced by the stallion and his initial semen quality). Fig. 4. Schematic presentation of the different factors affecting equine fertility. The proportions of the different factors are arbitrary. However, note how semen quality (in green as determined by the stallion) is influenced by management when performing the analysis (pink border). Vice versa, semen handing to prepare the AI doses is influenced by the initial sperm quality (smaller volumes of highly concentrated semen are easier to process). The influence of “management” on determination of the sperm quality should be minimal and standardized, facilitating comparison. Correct analysis of the semen will also point out the weaknesses in the semen preparation more rapidly, which will result in increased sperm management. This will be discussed in the next section, where we evaluated the effect of centrifugation on semen quality in horses. Mare Management Semen handling Semen quality Stallion 187
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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 (
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References CHAPTER 3.3 Arunvipas P.
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3.4.1. Abstract CHAPTER 3.4 In this
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CHAPTER 3.4 therefore in these expe
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CHAPTER 3.4 The averages obtained a
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3.4.4.2. Experiment 2 CHAPTER 3.4 F
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References CHAPTER 3.4 Arunvipas P.
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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 and 180: CHAPTER 4.2 Sperm yields markedly d
Page 181 and 182: References CHAPTER 4.2 Barth A.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: CHAPTER 5.1 al., 2007) showing a cl
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
Page 230 and 231: BIBLIOGRAPHY ABSTRACTS AND PROCEEDI
Page 233 and 234: FUNDAMENTAL ASPECTS OF CRYOPRESERVA
Page 235 and 236: ADDENDUM I presence of these CPAs a
Page 237 and 238: ADDENDUM I Fig. 3. Theoretical curv
Page 239 and 240: ADDENDUM II DETAILED LOADING TECHNI
Page 241: In m’n hoofd is alles heel eenvou
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