22/11/2006
Evaluation of the Quality of Deep-Frozen Semen and Selection of an Effective Method of Goat Sperm
Preparation for In Vitro Fertilization
By
W.P.D.K. FERNANDO
Introduction Objectives Semen Quality Analysis Selection of a Sperm Preparation method Results & Discussion Conclusion Acknowledgements End
CONTENT
INTRODUCTION
Production & Productivity of Local Goat
Demand for kids, meat & milk HIGH & ESCALATING
Supply ?
Reasons Lack of good quality breeding materials Selling of breedable or pregnant she goats for meat
Cont. INTRODUCTION
WHAT IS NEEDED Large amount of better breeding materials Sooner the possible At a lower cost*
Potentials Many sustainable farming systems AI & ET
Good but limited IN VITRO techniques have a higher potential
IVM IVF IVC
The success of the in vitro techniques depends on…Semen factors Semen quality (motility, abnormalities, live/dead ratio, etc.)
fresh or cryopreserved ? how long preserved ? semen of which breed ?
Sperm preparation why Sperm preparation ? which method ?
Classical sperm swim-up method Percoll gradient method Ficoll medium method Method of centrifugation on a discontinuous density gradient and
etc)
For Sri Lanka …
OBJECTIVES
To evaluate whether the quality of deep frozen semen is affected by the breed &/or preserved period
To select an effective method of sperm preparation for IVF in goat
To facilitate the implementation of IVF technology for breed improvement in local goat.
EX:1 EVALUATION OF SEMEN QUALITY 58 Cryopreserved semen straws of four cattle
Breeds Friesian Australian Milking Zeebu (FrAMZ) Jersey (Jr) Friesian (Fr) Friesian Sahiwal (FrSw) were selected
Classified into 5 age categories according to the semen straw preserved year > 20 years 19 – 15 14 – 10 9 – 5 <5
Quality Analysis 1
1.1 Motility Visual detection
1.2 Dead Sperm Percentage Eosin/Nigrosin staining method
1.3 Abnormalities William’s staining method
Ex1
EX:2 SELECTION OF A SPERM PREPARATION METHOD
Two sperm preparation methods were compared
Classical Sperm Swim-up Method A Simplified Sperm Swim-up Method
Ex2
Quality Analysis 22.1 Cleanliness of the semen
2.2 Motility Before & after preparation Visual Detection
2.3 Concentration Haemocytometer
Ex2
RESULTS & DISCUSSION
Means in a column with the same letter are not significantly different.Alpha = 0.05
Breed
Mean Motility
Age(yrs) Mean Motility
FrAMZ 24.50a >20 26.54a
Jy 39.06a 15-19 25.30a
Fr 41.88a 10-14 48.75a
FrSw 44.00a 5-9 40.89a
<5 45.25a
1.1 Sperm Motility
Table 1: Sperm Motility of different breeds and age categories
Ex1
Breed Mean
Death%
Age
(Yrs)
Mean
Death%
FrAMZ 65.63a >20 64.77a
Jy 54.56a 15-19 69.20a
Fr 56.12a 10-14 48.13a
FrSw 53.60a 5-9 56.16a
<5 46.38a
Table 2: Dead sperm percentages of different breeds and age categories
Means in a column with the same letter are not significantly different.Alpha = 0.05
1.2 Dead Sperm PercentageEx1
Correlation of sperm motility and Dead Sperm Percentage
A highly significant negative (Pearson) correlation (r = -0.882, P<0.01) between Sperm Motility and Dead Sperm Percentage.
Ex1
Table 3 - Sperm abnormalities according to breed:
Means in a column with the same letter are not significantly different.Alpha = 0.05
BreedAbnormalities
Head Mid Tail
FrAMZ 18.1a 12.1ab 17.9a
Jy 14.9a 8.4b 11.1b
Fr 18.3a 13.4a 10.4b
FrSw 17.5a 14.8a 8.9b
1.3 AbnormalitiesEx1
Table 4 - Sperm abnormalities according to storage time:
Means in a column with the same letter are not significantly different.Alpha = 0.05
Age Category
Abnormalities
Head Mid Tail
>20 16.8ab 9.5a 12.7a
15-19 17.2ab 9.4a 18.2b
10-14 13.6b 8.4a 8.9a
5-09 17.4ab 14.5b 11.2a
<5 20.8a 16.5b 11.1a
1.3 AbnormalitiesEx1
2.1 Cleanliness of prepared sperms Both methods resulted with slightly particulated
semen
Method
Swmup Sswmup
Initial motility 71a 72a
Final motility 90b 78a
Means with a same letter are notsignificantly different.Alpha = 0.05
2.2 Sperm Motility of both preparation methods
Ex2
Table 5 - Sperm concentration after sperm preparation
Method
Swmup Sswmup
Sperm Concentration
(x 106)4.5a 7.5a
swmup=swim-up methodsswmup=simplified swim-up methodMeans with the same letter are not significantly different.Alpha = 0.05
Ex2
CONCLUSION & SUGGESTIONS There is no effect of storage time and breed on motility
and dead sperm percentage but they affect on abnormalities. Other factors affecting motility & Dead Sperm Percentage need
to be evaluated
The simplified swim up method is as effective as the classical swim up method, but the former is more convenient and more economical. But this selection should be broadened to evaluate the
fertilization rate
ACKNOWLEDGEMENTS
Prof. D.P.S.T.G. Attanayake, Head, Dept. of Biotechnology, FAPMSL
is deeply acknowledged for his valuable encouragement & guidance.
Mr.D.V.S. de S. Gamage Head, Division of Animal Breeding,VRI Mr. A.S.B.Rathnayake Mr. P.A.B.S. Kumara,
Division of Animal Breeding, VRI are also deeply acknowledged for their valuable assistance during the research.
Mr.W.K.S. Amarasinghe and the staff Central Artificial Insemination Station (CAIS), Kundasale.
Dilip Fernando
IntroductionObjectives Ex 1Ex 2Results & DiscussionConclusion
Dilip Fernando
Abnormalities
Semen smear on glass slide Air dried and fixed in flame. Absolute alcohol for 4 min and air dried Washed d. water & with 96% ethanol Stained Carbolfuchsin-eosin solution (william’s
stain) for 10 min Washed with water Located on the stage of LMC
AI & ET vs IVF
we should not stay here we need develop more gene technologies spawning here and there thus we
thought of a far future IVF cant be used for gene manipulation which will be key
in future Immediately dead animals’ semen/oocytes can be used Even oocytes of new born she goats can be matured and
used in IVF
Top Related