Semen evaluation

Fertilizing ability of males depends on semen

quality, therefore, after collection of semen in

artificial insemination centers, the ejaculates

rottenly examined to assess the quality of semen

which refer to the fertilizing ability of males

After collection, the semen must be kept warm in
waterbath at 37°C and examined immediately to
evaluate the quality.

All the used buffers, slides, pipettes and tubes for

semen evaluation must be kept clean, dry and free
from spermicidal substances, and they must be placed
on a warm stage or stored in an incubator at 35-37°C.
 Collected semen should examined macroscopically and
microscopically,,,, and sometimes tests for metabolic activity of sperms

Macroscopic tests Microscopic tests

1- volume 1- motility

2- color and opacity a- mass motility

3- evaluation of hygienic quality b- individual motility

4- density and consistency 2- live and dead percentage
5- pH 3- sperm abnormalities

4- sperms concentration
 volume

Species Volume Species Volume

Bull 2-8 ml Stallion 50-150 ml

Buffalo-bull 3-6 ml Boar 150-250 ml

Ram 0.5- 1 ml Dog 10 ml

Camel – bull 2-12 ml

 pH
Species Initial pH Species Initial pH

Bull 6.4-6.9 Stallion 7-7.8

Buffalo-bull 6.4-6.9 Boar 6.9-7.9

Ram 5.9- 6.3 Camel-bull 7.6-7.8

Dog 6-6.8
Ejaculate with pus
 motility
Motility is the most often used criterion for routine semen
evaluation due to the simplicity of the evaluation technique, and
because it gives good indicator about viability of sperms in the
ejaculate.

fertile bulls usually produce semen with highly motile sperms, so in

artificial insemination centers, the mass and individual motility
should be measured for every ejaculate to estimate male fertility
 Mass motility
the strength of the mass motility (wavy motion) is varied
according to the sperm cell concentration, the number of motile
sperms and the strength of their motility.

A drop of undiluted semen (raw semen) placed on dry warm

slide, and examined under 4 or 10 X.
Wave motion Descriptive numerical
scale

No waves, sperm are immotile Very poor 0

poor 1
No waves, weak rotatory movement
fair 2
No waves, strong rotatory movement (< 50% motile sperms)
good 3
Slow wavy motion (50-80% motile sperms)
Very good 4
Rapid distinct wavy motion (~90% motile sperms)
excellent 5
Very rapid vigorous wavy motion with eddies (>90% motile)
Very active sperms(90-100%) of sperms are very active… an indicator of excellent fertile
male… notice the formation of eddies … (degree 5 of mass motility)
Degree 4 of mass motility… about 90% of sperms with very good mass motility… the
motion forms little waves of eddies
Degree 3 of mass motility… slow little waves without eddies (50-80% of sperms are
motile)
Degree 2 of mass motility… less than 50% of sperms are motile… no waves
Degree 1 of mass motility… very weak mass motility due to poor movement of
sperms…
Degree 0… immotile sperms
 Important notes
●Bull ejaculate should be good in descriptive scale
(number 3) to be acceptable for processing in AI centers.

●No mass motility in stallion or boar semen, or in bull

semen collected with electroejaculator or massage
techniques.
 Individual motility
estimated by mixing one drop of raw semen with two drops of 2.9% sodium citrate solution (or
warm saline 0.9%) on a warm slide at 37˚C, then the mixture covered by cover slide and examined
under light microscope at 40X magnification.
Motility scored on the basis of the percentage of sperms with normal forward progressive
movement, whereas those showing circling movements or those oscillating at one place were
regarded as immotile.
Progressive motility may define as the movement of the sperm from one point to another in a
straight line.

Individual motility Descriptive value %motile sperms

Very poor 0-20

of sperms motile 1/5
poor 20-40
of sperms motile 2/5
fair 40-60
of sperms motile 3/5
good 60-80
of sperms motile 4/5
Very good 80-100
of sperms motile 5/5
Weak individual motility,, about 20%
 Measurement of motility and concentration by Computer Assisted Sperm Analyzer (CASA)

Good individual motility with high Weak individual motility with

concentration low concentration
 Live and dead percentage
this can be done by staining the semen with vital stains such as
eosin-nigrosin stain, that cannot stain the live sperms but stains the
dead sperms (because of the destruction of dead sperm walls), so the
live sperms remain unstained or white, while the dead sperms takes the
red stain. Eosin-negrosin stain can be prepared by mixing (0.6% eosin,
5% nigrosin, 2.9% sodium citrate). One drop of semen is added to
several drops of eosin-nigrosin mixture on dry warm slide, and after
gentle mixing leave for about one minute. Make a smear from the
mixture on a clean slide and dry it, then examine under oil immersion
lens (100X). The percentage of dead and abnormalities were calculated
after counting 200 sperms.
Upper: live sperm; take no stain
lower: dead sperm; take red stain