chapter 3 - RiuNet

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STANDARDIZATION OF SPERM MOTILITY EVALUATION frequency (BCF; beats/s), defined as the average rate at which the curvilinear sperm trajectory crosses its average path trajectory. Spermatozoa were considered immotile if their VCL was 50% motile cells. All trials were carried out using each of these motility classes (except the ejaculate portion trial, in which only C-III class was used). 2.4 Effect of chambers and magnification lens Different tools can be used for sperm motility evaluation by CASA systems. In this trial, two commercially available chamber models, the ISAS D4C20 disposable chamber (20 µm deep; Proiser R+D) and the Makler reusable chamber (10 µm deep; Sefi Medical Instruments) and two magnification lenses (10x versus 20x in a Nikon E400 microscope, negative phase contrast) were tested. 2.5 Effect of frame rate To assess the effect of frame rate on the system’s ability to describe sperm motion, sperm quality parameters at 20, 30, and 60 fps (Hz) were compared. With the aim of avoiding variations between replicates within the same sample, the original file, captured at 60 fps, was manually modified using video analysis software removing one or two frames from every three original ones within each video file, as such obtaining files of 30 or 20 fps, respectively. 2.6 Effect of ejaculate portion and post activation time Sperm samples were collected in two portions: the first collectable millilitre (1st mL) was retrieved in a test tube, and the rest of the sperm was collected in another test tube. Sperm quality parameters of C-III class samples were also measured at different postactivation times (30, 60, and 35
CHAPTER 1 90 seconds), with the aim of assessing the effects of the differences in time from the sperm activation event. 2.7 Statistical analysis The mean and standard error (SE) were calculated for all the sperm quality parameters. Shapiro-Wilk and Levene tests were used to check the normality of data distribution and variance homogeneity, respectively. The one-way analysis of variance (ANOVA) and Student's t-test were used to analyze data with normal distribution. Significant differences between postactivation times were detected using the Tukey multiple range test (P
Page 2 and 3: SPERM PHYSIOLOGY AND QUALITY IN TWO
Page 4 and 5: ACKNOWLEDGEMENTS Muchos años han p
Page 6 and 7: aunque tiene una alcachofa en la ca
Page 8 and 9: TABLE OF CONTENTS SUMMARY 1 RESUMEN
Page 10 and 11: SUMMARY SUMMARY The conservation st
Page 12 and 13: RESUMEN RESUMEN Las especies objeto
Page 14 and 15: RESUM RESUM Les espècies objecte d
Page 16: GENERAL INTRODUCTION 7
Page 19 and 20: GENERAL INTRODUCTION who postulated
Page 21 and 22: GENERAL INTRODUCTION 2. Sperm physi
Page 23 and 24: GENERAL INTRODUCTION Finally, the a
Page 25 and 26: GENERAL INTRODUCTION 3. Sperm quali
Page 27 and 28: GENERAL INTRODUCTION in fish (Kime
Page 29 and 30: GENERAL INTRODUCTION However, for s
Page 31 and 32: GENERAL INTRODUCTION future applica
Page 33 and 34: GENERAL INTRODUCTION With regards t
Page 36: OBJECTIVES The chapters presented b
Page 40 and 41: STANDARDIZATION OF SPERM MOTILITY E
Page 56: CHAPTER 2 Study of the effects of t
Page 59 and 60: CHAPTER 2 reducing the pressure on
Page 61 and 62: CHAPTER 2 ongoing task in order to
Page 63 and 64: CHAPTER 2 To measure sperm density,
Page 65 and 66: CHAPTER 2 Spermiating males (%) 100
Page 67 and 68: CHAPTER 2 100 80 60 I II III IV A 4
Page 69 and 70: CHAPTER 2 3.2 Hormonal treatments P
Page 71 and 72: CHAPTER 2 With regard to the sperm
Page 73 and 74: CHAPTER 2 Relative volume (%) 80 I
Page 75 and 76: CHAPTER 2 In this respect, it is we
Page 77 and 78: CHAPTER 2 treatment to that of the
Page 79 and 80: CHAPTER 2 Our results demonstrate t
Page 82 and 83: ROLE OF IONS IN THE SPERM ACTIVATIO
Page 92: CHAPTER 4 Study of pufferfish (Taki
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CHAPTER 4 2002) so Takifugu niphobl
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CHAPTER 4 carrying out sperm collec
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CHAPTER 4 Table 1. Activation media
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CHAPTER 4 TM (%) 100 A a a Und-PD a
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CHAPTER 4 TM (%) 80 60 40 20 0 A a
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CHAPTER 4 TM (%) 80 A ASW100 GLU 60
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CHAPTER 4 although the activation m
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CHAPTER 4 dilution ratio and an opt
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CHAPTER 4 2006). Several studies in
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CHAPTER 4 5. Conclusions Some concl
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RELATIONSHIP BETWEEN SPERM MOTILIY
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GENERAL DISCUSSION 1. Improvements
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GENERAL DISCUSSION profiles induced
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GENERAL DISCUSSION more studies on
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GENERAL DISCUSSION sperm motility p
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CONCLUSIONS 133
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CONCLUSIONS vii. The coefficients o
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REFERENCES Aarestrup K, Økland F,
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REFERENCES Beirão J, Cabrita E, P
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REFERENCES Cabrita E, Robles V, Cu
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REFERENCES De Vos A, Van De Velde H
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REFERENCES Gallego V, Pérez L, Ast
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REFERENCES sperm competition and fe
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REFERENCES Liu QH, Li J, Xiao ZZ, D
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REFERENCES Mortimer ST. Effect of i
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REFERENCES Peñaranda DS, Marco-Jim
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REFERENCES Rurangwa E, Volckaert FA
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REFERENCES Tesch F. Telemetric obse
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REFERENCES Wong PYD, Lee WM, Tsang
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