EXAMINATION AND

EVALUATION OF
SEMEN
PLACE OF
EXAMINATIO • Artificial Insemination Laboratory and
N AND • Reproductive Physiology Laboratory
EVACUATION
of SEMEN
 • Semen should not be
• – Exposed to direct sunlight,
DURING • - contact with metal,
SEMEN • - mixed with chemicals, urine, water etc.,
• - shaken vigorously and often
EXAMINATI • Must be careful and as fast as possible without
ON compromising accuracy
• The tool must be clean and sterile, temperature
37oC
 MACROSCOPIC

TYPES OF MICROSCOPIC
SEMEN
EXAMINATION BIOLOGY

BIOCHEMISTRY
MACROSCOPIC
EXAMINATION
VOLUME

CONSISTENCY

COLOR

SMELL

pH
MACROSCOPIC
EXAMINATION

VOLUME

- VIEWING SCALES IN COLLECTION TUBES

- Average cattle2 4ml and sheep 1ml
MACROSCOPIC
EXAMINATION

consistency
- TILT THE COLLECTION TUBE THEN STRAIGHTEN,
LOOK AT THE WALL OF THE TUBE, IF IT IS SLOW
DOWN THEN IT IS VISCOUS
- Normal generally medium to viscous
MACROSCOPIC
EXAMINATION

COLOR

See the color of semen from milky

white to yellow (Cow), white to
cream (Sheep) sperma domba
MACROSCOPIC
EXAMINATION

SMELL

Cow  like milk

Sheep  prengus
MACROSCOPIC
EXAMINATION

pH

- pH meter
- Ph paper
- Normal pH 6.4 – 6.8 (Cow and Sheep)
MICROSCOPIC EXAMINATION

MASS MOVEMENT

INDIVIDU MOVEMENT

CONCENTRATION

THE NUMBER OF LIVE AND DEAD SPERMATOZOA

NUMBER OF SPERMATOZOA ABNORMALITIES (MORPHOLOGICAL)

MICROSCOP
IC • The movement of several spermatozoa cells
EXAMINATI together thus forms a wave
• Reflects the impulse and concentration of
ON spermatozoa cells
MASS • 37o C in order to obtain optimal sperm cell
movement
MOVEMENT
MICROSCOP HOW
IC 1) Put one drop of semen on the glass of the
object,
EXAMINATI 2) Observe mass movement under
ON a microscope at 100 times magnification.
MASS
MOVEMENT

+++ ++
MICROSCOP
IC • Video of mass movement +++ ; ++ and + in
cows
EXAMINATI • Video of mass movement +++ ; ++ and + in
ON cows/sheep

MASS
MOVEMENT
MICROSCOP
IC • Movement per individual spermatoazoa
EXAMINATI • It is important to reach the egg (ovum)
present in the fallopian tubes
ON • 37o C in order to obtain optimal sperm cell
INDIVIDUAL movement
• Should be done as soon as the semen is
MOVEMENT accommodated
S
MICROSCOP
IC CARANYA
• 1). Letakkan satu tetes semen di atas gelas
EXAMINATI obyek, tambahkan satu tetes atau 1 : 10 atau
ON 1 : 100 larutan NaCl fisiologis campur sampai
homogen.
INDIVIDUAL • 2). Tutuplah gelas obyek dengan gelas
MOVEMENT penutup dan
• 3). Dilakukan pemeriksaan di bawah
S mikroskop dengan pembesaran 400 kali.
MICROSCOP VIDEO
IC • 60/3  60% progressive motile at fast speed
• 55/3  55% progressive motile at fast speed
EXAMINATI • 30/3  30% progressive motile at fast speed
ON • 45% 3+.mpeg
INDIVIDUAL • 60% prog 3 ind.mpeg
• 80 % progresif 3+ ind.mpeg
MOVEMENT • 90 % individu 3+.mpeg
S
MICROSCOP
IC METODE
EXAMINATI • RUSIA

ON • HAEMACYTOMETER PAPAN THOMA/NEUBAUER

IMPROVED
CONCENTR • SPEKTROFOTOMETER

ATION • ALAT BLOM

MICROSCOP EVALUATION:
IC DENSUM (D)  DISTANCE LESS THAN ONE
EXAMINATI HEAD SIZE SPERMATOZOA (>1000M/ml)
SEMI DENSUM (SD)  DISTANCE OF MORE
ON THAN ONE HEAD SIZE OF SPERMATOZOA
CONCENTR (500M-1000M/ml)
RARUM (R)  DISTANCE IS ALMOST EQUAL TO
ATION  ONE SIZE OF SPERMATOZOA (< 500M/ml)
RUSSIAN AZOOSPERMIA (A)  NO OR FEW
SPERMATOZOA
MICROSCOP
IC
EXAMINATI
ON METHOD
CONCENTR THOMA BOARD
ATION NEUBAUER BOARD IMPROVED

HAEMAC
YTOMETER
MICROSCOP HOW
1). First sucked semen using an erythrocyte pipette
IC up to 0.5.

EXAMINATI 2). Then add diluent (NaCl 3% methylen

blue/eosin) until the number 101. So that the
ON amount of diluent: 100/0.5 = 200 times, because
the liquid in the pipette up to 1.0 does not mix.
CONCENTR 3). Bend the tip of the suction rubber and shake the
pipette in a motion to form a figure eight several
ATION times until the solution inside is homogeneous.
HAEMAC 4). Furthermore, before the examination, dispose
of 2-4 drops of solution to remove the liquid in the
YTOMETER pipette that did not mix earlier.
5). Drip the above solution on Thoma's counting
board through one side of the cover glass.
MICROSCOP
IC
EXAMINATI HOW TO CALCULATE
Five (5) large squares consisting of 4
ON diagonal squares and 1 square at the
other corner.
CONCENTR So what counts is 5 x 16 small squares =
80 small squares.
ATION In thoma counting board there are 400
small squares in all.
HAEMAC If the number of spermatozoal cells in
80 small squares = y, so in 400 small
YTOMETER squares (0.1 mm3) semen contains
400/80 x 200 x Y.
Means in 1 mm3 of semen contains =
400/80 x 200 x 10 x Y = 10000 Y. So, if Y
= 100 cells then in 1 mm3 of semen
there are 10000 x 100 = 1000 000
spermatozoa cells / mm3.
MICROSCOP
HOW
IC 1). Turn on the spectrophotometer for 10
EXAMINATI minutes
ON 2). Setting the wavelength (546 nm) 3). Insert
physiological NaCl into the cuvette as much as
CONCENTRATI 4 ml and then tera in the spectrophotometer
ON 4). Take the cuvette, add 40 microliters of
semen, stir until homogeneous slowly.
SPECTROPHOT
5). Put it again into the spectrophotometr then
O-METER see the results on printing paper.
MICROSCOPI HOW
C • Take a clean glass object and if the former needs
to heat or remove it with alcohol to free the fat
EXAMINATIO layer.
N • On the glass object is given one small drop of
semen and one large drop of Eosin nigrosin
THE NUMBER solution beside it.
OF LIVE AND • Mix the dye with semen until homogeneous.
DEAD • Make a thin knead preparation and dry on
the flame (this process should be completed
SPERMATOZOA in a maximum of 15 seconds).
• See under a microscope with 400X
magnification.
 EVALUATION
• SPERMATOZOA LIVE HEAD UNCOLORED;
MICROSCOPIC • DEAD SPERMATOZOA COLORED HEAD
EXAMINATIO
N
THE NUMBER
OF LIVE AND
DEAD
SPERMATOZO
A

WHAT PERCENT IS ALIVE?

 EVALUATION
• SPERMATOZOA LIVE HEAD UNCOLORED;
MICROSCOPIC
• DEAD SPERMATOZOA COLORED HEAD
EXAMINATIO
N
THE NUMBER
OF LIVE AND
DEAD
SPERMATOZO
A
 EVALUATION
• SPERMATOZOA LIVE HEAD UNCOLORED;
MICROSCOPIC • DEAD SPERMATOZOA COLORED HEAD
EXAMINATIO
N
THE NUMBER
OF LIVE AND
DEAD
SPERMATOZO
A
 EVALUATION
• SPERMATOZOA LIVE HEAD UNCOLORED;
MICROSCOPIC
• DEAD SPERMATOZOA COLORED HEAD
EXAMINATIO
N
THE NUMBER
OF LIVE AND
DEAD
SPERMATOZO
A
 • PRIMARY
MICROSCOPI • SECONDARY
C
EXAMINATIO
N
NUMBER OF
SPERMATOZOA
ABNORMALITI
ES
MICROSCOPI • PRIMARY
• SECONDARY
C
EXAMINATIO
N
NUMBER OF
SPERMATOZOA
ABNORMALITI
ES
 RESISTANCE
BIOLOGICAL
TEST
EXAMINATI
ON
CATALASE TEST
BIOLOGICAL
EXAMINATI HYPOTONIC SOLUTION
ON
RESISTANCE DEFENDING AGAINST HYPOTONIC
TEST SOLUTIONS
 How

BIOLOGICAL
Prepare an Erlenmeyer tube (100-200 ml), with a pipette.
EXAMINATI 0.02 ml of semen was taken into the Erlenmeyer flask then
ON added 10 ml of 1% NaCl, then the solution was stirred
slowly and examined the motion of spermatozoa cells
RESISTANCE under a microscope with a magnification of 100 times.
With each addition of 10 ml of 1% NaCL is attempted
TEST continuously and will be stopped after many spermatozoa
cells appear whose movements only rotate.
This process must be done at a temperature of 20 – 25o C
and the entire experiment must be completed in 5
minutes..
 DETERMINING SEMEN HYGIENE
BIOLOGICAL BLOOD CELLS AND BACTERIA IN
EXAMINATI SEMEN PRODUCE THE ENZYME
CATALASE THAT CAN SEPARATE
ON OXYGEN FROM A SOLUTION OF WATER
CATALASE PEROXIDE (H2O2)
TEST THE CATALASE NUMBER IS THE
AMOUNT OF AIR FORMED ON TOP OF
THE LIQUID IN THE CATALASE TUBE
 DEHYDROGENIZATION
BIOCHEMIC TEST
AL
EXAMINATI
ON ASCORBINE ACID
LEVEL TEST
BIOCHEMIC This test principle is based on the length of
time from the semen to change the blue
AL color of the methylen blue solution.
EXAMINATI From the speed of changing the color of
methylen blue can be determined the
ON fertility level of semen.
DEHYDROGE
NIZATION
TEST THE FASTER IT CHANGES BLUE, THE MORE
FERTILE THE SEMEN WILL BE
BIOCHEMIC Each 100 ml of fertile
AL
EXAMINATI semen contains 3 – 8
ON mg of ascorbin acid
ASCORBINE
ACID LEVEL Less fertile <3 mg
TEST
ascorbic acid,
or >8 mg
 Breed of animals : Sheep
REPORT Age
Animal name
:
:

1. MACROSCOPIC EXAMINATION Parameters Result Normal Disorder Description

Volume
Consistensy
Color
Smell
pH

Noted:
- Results column created simulation
- The Normal column can be searched from readings both
Indonesian and Foreign
 Parameters Result Normal Disorder Description
REPORT Mass movement
Individual
movements
2. MICOSCOPIC EXAMINATION Concentration
(Russia)
Concentration
(Thoma)
Concentration
(spektrofotometer)
% alive
% dead
% Abnormalities

Noted:
- Results column created simulation
- The Normal column can be searched from readings both
Indonesian and Foreign
 RESISTANCE
REPORT
3. TEST
BIOLOGICAL
EXAMINATI • - PRINCIPLE
ON • - PROCEDURE
• - EVALUATION
REPORT

• COLLECTED IN ARTIFICIAL INSEMINATION

LECTURER ROOM A314 3RD FLOOR
• AT THE LATEST TO BE COLLECTED TOMORROW AT
13:00
• DON'T FORGET THE REPORT BOOK IS NAMED, NIM
AND CLASS
THANK
YOU