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LABORATORIUM BIOSAINS
UNIVERSITAS BRAWIJAYA MALANG
BBIB-SiINGOSARI MALANGBIOSAINS
THE QUALITY AND Ca +2 INTENSITY CHARACTERS OF
LOCAL INDONESIAN GOAT SPERM AFTER FREEZING BY
NONCONVENTIONAL METHODS OF CRYOPRESERVATION
LPDP-RISPRO , KEMENKEU, Rep. Of Indonesia
Contract No. : PRJ-1047/LPDP/2015
Dr. Gatot Ciptadi
Fac. Of Animal Husbandry, Brawijaya
University, Indonesia
Supported by:
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Artificial Insemination (AI)
Implementation.
There are still very many types of
livestock / animals have not been
frozen (sperms)
Potential alternative technologies
sperm freezing and conservation of
genetic materials for both research
and commercial purposes.
Goat Population East Java:
3.178.197 heads, Indonesia:
18.879.596 heads
spermatozoa
INTRODUCTION
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Research program : 2 YEARS
Year I : Goat
Senduro
Lumajang- East
Java
Year II: Sheep: DEG,
Sapudi Island , East
Java, Indonesia
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Location
Method of ReserachRealization of Cell Bank (Sheep and Goats)
Periode Actifities Goal and Targets Lokasi dan sampelpercobaan
Year I:
•Research
•Production
•Aplication
•Marketing
Male Selection
Chromosom and DNAAnalysis
Sperm Freezing: Production andin vitro test of quality
Sperm freezing, qualitycontrol, production andstock.
Genetic quality information
BBIB Singosari,
LSIH and LaboratoriumBiosains UB
Research Station Fac. OfAnimal Sci. UB.
Small holder farm
Year II:
•Research
•Production
•Aplication
•Marketing
Aplication of AI
Ferlitity test ( lab and Field trial)
Commercial sperm production
Promotion
AI BBIB Singosari,
LSIH and LaboratoriumBiosains UB
Research Station Fac. OfAnimal Sci. UB.
Small holder farm
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
The Goals of Research
Realizing a frozen cell bank ( spermatozoa ) goat -
sheep for conservation and commercialization
purposes of:
Year I: Goat Senduro ( Lumajang )
Year II : Fat Tailed Sheep ( Sapudi Island )
Utilization stock products frozen cell ( -196 0C ) for
research and AI applications
Important : Feasibility study of the conservation of
genetic material from a technical , economic and
scientific aspect as well as to anticipation of the
possibility of increasing development of
international trade cells .The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Methods Lab experiment method . and Application
Field ,Producing frozen cell ( spermatozoa )
quality (genetic , viability , fertility , productivity
, specific) Spermatozoa livestock / animals
native, superior criteria , specific or rare .
Products stored frozen cells -196 0C , straw
0:25 ml . animals used previously have
genetic screening ( chromosomes and DNA )
to guarantee the quality and free of defects .
The production process and the freezing of spermatozoa in BBIB
Singosari ( ISO 17025 : 200 / ISO 9001/2008 ) based on ISO 2008 .
Test genetic / chromosomal DNA carried in LSIH - UB and UB
Biosciences Laboratory UB ( ISO 17025 : 2008 and ISO 9001 : 2008 )
. Frozen cell product testing laboratory fertility in vitro followed by IB
application tests in the field.
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Research Supporting (Field Trials)
The 12 students in the field in collaboration with
the Department of Livestock Lumajang, with the
selection and measurement of vital statistics
phenotypic characters, total of 900 individuals (
including 249 males ) .
students who study: freezing and testing the
quality of sperm goats and Senduro with
vitrification method .
Males were high selected ( selected 1.0 % ) .
Tim BBIB and UB : Measuring Vital statistics
candidate male
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
LABORATORY RESEACH RESULT
Sperm quality In vitro and Genetic Quality.
Selected goats : analyzed normal
genetically ( chromosomally normal )
the quality of spermatozoa (fresh) : motility
more than 80% of fresh semen and more
than 40 % post- thawing motility ( PTM ) in
accordance with ISO 2008 .
Genetic analysis has been completed and
indicates that the goat Senduro and PE is
normal based on the analysis of
chromosomes ( 2N = 60 , X = acrosentris , Y
= metasentris ) , a DNA analysis on the
stages of DNA isolation followed by PCR for
gene identification GHThe 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
RESEARCH RESULTS(1). Male Goats have been testedfor genetic , spermatozoa quality ,
libido and test post-thawing motility (PTM and Implemented AI resulted
in high lever result ( 86.67 % pregnancy rate)
(2). Research Genetic analysis of goats selected (chromosome and
DNA analysis has been completed).
Chromosome analysis showed the normal : number and structural
(3). Initial production of frozen semen and test quality goat (Senduro)
already done, freezing still continue to be performed
(4). Goat sperms Ca++ intensity is different and
varied beetwen fresh and Freezing sperm.
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
Figure 1. Different profile of Calcium intensity of local goat sperm :
(a).Fresh sperm
(b). Freezing sperms (post thawing) of local Indonesian Goat.
Analyses by Confocal Laser Scanning Microscope (CLSM : 400, Z.4 , 3 D).
The Calcium Intensity profile
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
C o n c l u s s i on1.Sires are selected based on pedigree and a high level of selection
intensity , phenotypic and vital statistics standard
2. Sperm quality is considered (ISO 2008)
3. The calcium intensity pattern is varied more and higher
in fresh sperm than freezing sperm. The local goat sperm
frozen by non conventional could be used for the
application of AI. However, this method needs to be
enhanced using diluents and cryoprotectant more
appropriate.
Suggestions
It was important to study the relationship between the
relative intracellular calcium intensity both fresh and
freezing semen with their fertility in vitro.
.The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN
FOR MORE INFORMATION:
www.bankselgamet.com1. . LPDP-RISPRO. Kemenkeu
(FUND MANAGEMENT
INSTITUTE OF EDUCATION
MINISTRY OF FINANCE No.
Agreement : PRJ - 1047 / LPDP
/ 2015
2. BBIB Singosari Malang
3. Goat Breeders Assoc. Etawah
Jaya , Lumajang
4. The District Livestock Service
Office . Lumajang
5. All Goat Small Holder Farm/
Breeders Senduro districts .
The 17 th AAAP ANIMAL SCIENCE CONGGRESS
22-25 August 2016, FUKUOKA, JAPAN