the quality and ca +2 intensity characters of … · the production process and the freezing of...

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



Research program : 2 YEARS

Year I : Goat

Senduro

Lumajang- East

Java

Year II: Sheep: DEG,

Sapudi Island , East

Java, Indonesia



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 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


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.



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



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


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.



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



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


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 .



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