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STANDARDIZATION OF SPERM MOTILITY EVALUATION Moreover, regarding change in movement parameters after sperm activation, MOT was the most affected factor, displaying a progressive decrease in the percentage of motile cells after the activation time. Usually in marine and freshwater species, most of the sperm traits used to characterize motility decline within tens of seconds to a few minutes, depending on the species, and this general decrease leads to an eventual full arrest of spermatozoa by ATP consumption (Cosson et al., 2008b). In the case of European eel, in addition to the reduction in motility, a decrease in the percentage of fast spermatozoa was also evident over time. As such our results suggest that sperm analysis in European eel sperm should be performed within the first 60 seconds after activation. 5. Conclusions Computer assisted sperm analysis systems are useful tools for carrying out studies about spermatozoa kinetic parameters in fish species. However, some questions have emerged regarding sperm sample evaluations, and as such several procedural and technical settings should be standardized and validated before comparing results obtained by different laboratories. In this study, we have assessed different technical and biological settings to standardize the evaluation of sperm quality in the European eel and use it as a sperm model for teleost fish. We have discovered that some protocol variables in sperm analysis using CASA software (ISAS v1) can affect the measurement of eel (fish) sperm quality parameters. Notably, neither the type of chamber nor the ejaculate portion affected the sperm quality parameters, suggesting that either type can be used for sperm evaluation in European eels. Finally, to carry out a suitable analysis on sperm quality parameters in European eel, we suggest a few recommendations regarding its application: (1) use the lowest available magnification lens, with the aim of avoiding a large spread in data; (2) use the highest available frame rate, with the aim of obtaining the most “real trajectory” of the spermatozoa; and (3) perform the analysis within the first 60 seconds after activation. 45
CHAPTER 1 Acknowledgements This study was funded from the European Coµmunity’s 7th Framework Programme under the Theme 2 “Food, Agriculture and Fisheries, and Biotechnology,” grant agreement 245257 (Pro-Eel), and Generalitat Valenciana (ACOMP/2011/229). D.S.P. and P.C.F.C. have postdoctoral grants from UPV (CE-01-10) and PAC-EMBRAPA, respectively. I.M. and V.G. have predoctoral grants from Generalitat Valenciana and Spanish MICINN, respectively. The authors thank the Proiser R+D, S.L. team, who performed the task of fractioning the original sequences for the experiment described in section 3.2. 46
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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
Page 95 and 96: CHAPTER 4 2002) so Takifugu niphobl
Page 97 and 98: CHAPTER 4 carrying out sperm collec
Page 99 and 100: CHAPTER 4 Table 1. Activation media
Page 101 and 102: CHAPTER 4 TM (%) 100 A a a Und-PD a
Page 103 and 104: 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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