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CHAPTER 4.2 166 4.2.3.4. Experimental design Each of the 22 ejaculates was processed as described above. After re-dilution in BotuCrio, the sperm yield expressed as % was calculated [(final volume of in BotuCrio extended semen × 100×10 6 sperm/mL) divided by 3000×10 6 initial sperm per treatment multiplied by 100] and an aliquot of semen from each treatment was analyzed with CASA and a morphology smear was prepared prior to cryopreservation. Frozen semen was thawed and TM and PM were analyzed using CASA, while DFI, MI, AR and morphology were determined as well. 4.2.3.5. Statistical analysis sperm quality parameters All data were percentages and were transformed by dividing them by 100 followed by calculating the arcsine of the square root of that value, in order to obtain a normal distribution as assessed by QQ-plots. The possible effect of “Treatment” (cushion vs. Androcoll-E) as the main variable of interest, on the different parameters (TM, PM, NM, AH, DFI, MI, AR, and yield, respectively) was studied. Therefore, multilevel models were fitted in MlwiN 2.02 (Centre for Multilevel Modeling, Bristol, UK) with the eight different parameters as outcome variables and “Treatment” as fixed effect and “Stallion” as random effect to adjust for clustering of ejaculates within stallion. As well, “Freezability” (good vs. poor, see above) and the interaction term “Treatment” × “Freezability” were included in the model. Non-significant terms (p>0.05) were excluded from the full models. accepted straws and number of AI doses The association between “Treatment” and “Number of accepted straws” was studied using Chi 2 on 2×2 tables for poor and good freezing stallions using Win Episcope 2.0 (CLIVE, Edinburgh, UK). Breslow-Day test indicated interaction was present between “Freezability” and “Treatment” allowing only for stratum-specific (poor vs. good freezing stallions) conclusions. Odds ratios with 95% confidence interval were calculated to visualize the strength of the associations. As well, Chi² on 2×2 table was used to test whether “Number of doses” was related to “Treatment” for good and poor freezing stallions. Significance was set at p
4.2.3. Results 4.2.3.1. Sperm quality parameters CHAPTER 4.2 Four stallions were considered to have a good freezability whereas the other four stallions were poor freezers. Good freezers had a significant higher pre-freeze TM and PM, and a higher post- thaw TM and MI compared to poor freezers (Table 1). The other parameters did not differ significantly between good and poor freezers. 100 80 60 40 20 0 A. Pre-freeze progressive motility (%) B. Yield (%) poor freezing stallions Fig. 2. Pre-freeze progressive motility and yield (with SD-bars) of stallion sperm following cushioned centrifugation or selection through Androcoll-E (good freezing vs. poor freezing vs. all stallions) of the stallion’s ejaculates. Analysis of the pre-freeze samples showed a significant improvement in sperm quality parameters for Androcoll-E treated sperm, although this was borderline non-significant for TM (Table 1). However, sperm yield was markedly lower compared to the cushioned group. Analysis of the post thawed samples revealed an overall increase in sperm quality for the Androcoll-E treated samples (Table 1). Cushion Androcoll-E good freezing stallions all stallions 120 100 80 60 40 20 0 poor freezing stallions Cushion Androcoll-E good freezing stallions all stallions 167
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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
Page 123 and 124: CHAPTER 3.3 The extended semen was
Page 125 and 126: CHAPTER 3.3 Table 1: Average values
Page 127 and 128: 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: CHAPTER 4.2 adding 600 µL of AO st
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 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
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CURRICULUM VITAE Maarten Hoogewijs
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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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