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CHAPTER 1.1 8 In the next paragraphs, the state-of-the-art concerning the analysis of (equine) semen is reviewed. Where possible, the comparison is made with the guidelines according to the World Health Organization (WHO). The WHO laboratory manual for the examination of human sperm and sperm- cervical mucus interaction was published for the first time in 1980. This guideline was developed in response to a growing need for the standardization of procedures for the examination of human semen. The manual has ever since provided global standards for human semen analysis and has been updated on numerous occasions based on new understandings and developments. In 2010 the fifth edition was published. 1.1.2 Gross evaluation of equine semen quality Following ejaculation the gel is separated from the remainder of the semen in the laboratory if no inline filter was applied during collection. The color and consistency of the semen are recorded. Normal semen should be white, grayish-white to slightly cream coloured and this provides a rough estimate of sperm concentration. Usually, larger volumes are more watery instead of a creamy consistency and this will be reflected in low sperm concentrations. A deviant color of the ejaculate may indicate the presence of admixtures, such as blood, urine or purulent material (Estrada and Samper, 2007; Binsko et al., 2011). The conventional semen analysis consists of the evaluation of semen volume, spermatozoal concentration, morphology and motility. Additional evaluations using more recent techniques might improve the predictive value of the examination (Varner, 2008). 1.1.3. Volume The volume of an ejaculate is most often determined by pouring the semen from the collection bottle into a tilted, pre-warmed graduated cylinder (Jasko 1992; Picket, 1993; Squires et al., 1999). This is in sharp contrast with human andrology, where it is advised to weigh the empty and filled container in order to calculate the volume based on the density of semen (WHO, 2010). For human samples, it is not recommended to aspirate or to decant the sample into a graduated cylinder. Anyhow, by decanting a part of the sample will get lost and sample losses have been reported to vary between 0.3 and 0.9 mL (Brazil et al., 2004; Iwamoto et al., 2006; Cooper et al., 2007). Obviously, these losses weigh heavily when handling small volume samples in human andrology with a total volume of about 4.0 mL, when compared to equine ejaculates in which a 10 times larger volume is most often present (Picket, 1993; Squires et al., 1999). In species with lower volume ejaculates (bulls, rams,…) losses due to decanting will be more important.
1.1.4. Concentration CHAPTER 1.1 According to the WHO manual (WHO, 2010), determination of the concentration of a human semen sample is preferably done using a 100 µm deep haemocytometer. The improved Neubauer haemocytometer is most commonly used. These evaluations are rather time consuming and as such, not easily applicable in production settings of animal AI companies where numerous ejaculates are processed. Hence, automated devices are commonly used in animal andrology, which is in contrast with practices in human andrology laboratories. Photometers Photometers (Fig. 1a) are probably the most frequently used devices for estimating concentration. Determination of concentration is based on the absorbance of a part of a light beam by the particles (spermatozoa) in a suspension. The alteration in light transmitted through a sample corresponds to the concentration of particles (Squires et al., 1999). As such, these instruments allow rapid measurements of concentration with a good accuracy as long as the concentration is within range (>100 and
Page 1 and 2: AUTOMATED AND STANDARDIZED ANALYSIS
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Page 7 and 8: A Area AI Artificial Insemination A
Page 9 and 10: PREFACE The first recorded case of
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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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4.1 Influence of different centrifu
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CHAPTER 4.1 4.1.2. Introduction 140
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CHAPTER 4.1 142 4.1.3.3. Semen proc
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CHAPTER 4.1 144 In the experiment 3
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CHAPTER 4.1 centrifugation (T1) and
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CHAPTER 4.1 (non CP vs CP) was sign
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35 B 80 A 33 beat cross frequency (
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CHAPTER 4.1 152 4.1.4.2. EXPERIMENT
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CHAPTER 4.1 Love and coworkers (200
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CHAPTER 4.1 Love C.C., Brinsko S.P.
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4.2.1. Abstract CHAPTER 4.2 The inc
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4.2.3. Materials and methods 4.2.3.
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CHAPTER 4.2 For the SLC group, 15 m
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CHAPTER 4.2 adding 600 µL of AO st
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4.2.3. Results 4.2.3.1. Sperm quali
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CHAPTER 4.2 The aforementioned “T
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CHAPTER 4.2 Sperm yields markedly d
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References CHAPTER 4.2 Barth A.D.,
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CHAPTER 4.2 Waite J.A., Love C.C.,
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CHAPTER 5 This thesis originated fr
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CHAPTER 5.1 Fig. 1. Schematic prese
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CHAPTER 5.1 higher than the current
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CHAPTER 5.1 al., 2007) showing a cl
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CHAPTER 5.1 treatment strategy for
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CHAPTER 5.2 supernatant allowing sp
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CHAPTER 5.2 such keeping the pellet
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CHAPTER 5.2 components of the semin
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Final reflections 5.4 Actual change
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CHAPTER 5 Dillon R.H., Fauci L.J.,
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CHAPTER 5 Suárez S.S., Osman R.A.
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SUMMARY technique that enables proc
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SUMMARY from stallions previously c
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SAMENVATTING beweeglijkheid (Hoofds
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SAMENVATTING de spermacellen gesele
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DANKWOORD Het obligatoire dankwoord
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DANKWOORD dankzij jou en ze zijn er
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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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