SEMEN

 Semen Composition
o Spermatozoa 5%
o Seminal fluid 60% to 70%
o Prostate fluid 20% to 30%
o Bulbourethral glands 5%
 The testes are paired glands in the scrotum that contain the seminiferous tubules for the secretion
of sperm
 The external location of the scrotum contributes to a lower scrotum temperature that is optimal
for sperm development
 Germ cells for the production of spermatozoa are located in the epithelial cells of the
seminiferous tubules.

STEPS:
1. Specialized Sertoli cells provide support and nutrients for the germ cells as they undergo
mitosis and meiosis (spermatogenesis).
2. When spermatogenesis is complete, the immature sperm (nonmotile) enter the epididymis.
3. In the epididymis, the sperm mature and develop flagella.
 The entire process takes approximately 90 days.
 The sperm remain stored in the epididymis until ejaculation, at which time they are propelled
through the ductus deferens (vas deferens) to the ejaculatory ducts.
 Ejaculatory ducts receive:
a) Sperm from ductus deferens
b) Fluid from the seminal vesicles (60-70%)
 Contain fructose and flavin
 Fructose – energy for motility
 Flavin - gray appearance of semen
1) Muscular prostate gland - aids in propelling the sperm through the urethra by contractions
during ejaculation.
2) Prostate gland – Produces approximately 20% to 30% of the semen volume which is acidic
fluid.
3) Coagulation and liquefaction of the semen following ejaculation - milky acidic fluid contains
high concentrations of acid phosphatase, citric acid, zinc, and proteolytic enzymes.
4) Bulbourethral glands – Contribute 5% of the fluid volume in the form of thick, alkaline mucus
that helps to neutralize acidity from the prostate secretions and the vagina.
 It is important for semen to be alkaline to neutralize, without this neutralization, sperm motility
would be diminished.

Specimen Collection
Missing portion:
First portion Second portion
(Most of the sperm are contained)
 sperm count will be decreased  sperm count is falsely increased
 pH falsely increased  pH is falsely decreased
 specimen will not liquefy  specimen will not clot
 semen volume is decreased
  Sexual abstinence of at least 2 days to not more than 7 days
o Prolonged abstinence tends to have higher volumes and decreased motility
o WHO: two or three samples be collected not less than 7 days or more than 3 weeks apart,
with two abnormal samples considered significant.
 Kept at room temperature and delivered to the
laboratory within 1 hour of collection.
 Awaiting analysis: 37 deg
 Label:
o patient’s name and birth date
o period of sexual abstinence
o completeness of the sample
o difficulties with collection
o times of specimen collection
o specimen receipt
Semen Analysis
 appearance
 volume
 viscosity
 pH
 sperm concentration and count
 motility
 morphology

Appearance Gray-white color Normal

Translucent
Musty odor
Low sperm concentration Infertile
Turbid Presence of white blood cells
Red coloration Presence of red blood cells
Yellow Urine contamination
Liquefaction 30 to 60 minutes after collection Normal
>60 minutes Deficiency in prostatic enzymes
>2 hours Induce liquefaction for analysis
using Dulbecco’s phosphate-
buffered saline or proteolytic
enzymes (e.g
alphachymotrypsin
or bromelain)

Jelly-like granules (gelatinous No significant

bodies)
Mucus strands Interfere
Volume 2-5 mL Normal
Extended abstinence Increase volume
Infertility Decrease volume
(improper function, primarily
 seminal vesicles)
Viscosity (consistency) Droplets Normal
Easily drawn into pipette
Clumped Abnormal
Highly viscous
Reporting 0 (watery) to 4 (gel-like)
Low, normal, or high
pH 7.2 – 8.0 Normal
Evaluated within 1 hour
Infection within the Increased
reproductive tract
Increased prostatic fluid Decreased
Ejaculatory duct obstruction
Poorly developed seminal
vesicles
Sperm count Sperm cells/ejaculate Normal: >40M/ ejaculate
Sperm Motility Forward, progressive movement Normal
Undiluted Proper examination
Well-mixed liquefied
20 HPO fields
Speed and direction
50% rating of 2.0 Normal
Computer-assisted semen Automated evaluation
analysis
Sperm Morphology Oval-shaped head Normal
Long, flagellar tail
Poor ovum penetration Abnormalities in head
Poor motility Affected parts are neckpiece,
midpiece, and tail abnormalities
Thinly smeared stained Evaluation
Oil immersion
Air-dried slides for 24 hours
200 sperm evaluated Abnormal
sperm percentage
Wright’s Giemsa Stain (lab preference)
Shorr
Papanicolaou
Kruger’s strict criteria evaluate sperm morphology
(not routinely performed but (measuring head, neck, and tail
recommended by WHO) size; acrosome
size; and of vacuoles)
>30% normal forms Routine criteria
>14% normal forms Strict criteria

Sperm motility can be evaluated at room temperature or 37°C, sample should be incubated at this
temperature and the preparation made with prewarmed slides and cover slips.
Acrosomal cap - half of the head and cover approximately two thirds of the sperm nucleus
Additional Testing
Sperm Vitality
Seminal Fluid Fructose
Decreased motility with normal
count
Test using eosin-nigrosin stain
Large proportion of vital but
immobile cells
High number of nonviable cells
and immotile
 Abnormalities of the seminal
vesicles
 Bilateral congenital absence
of the vas deferens
 Obstruction of the
ejaculatory duct
 Partial retrograde ejaculation
 Androgen deficiency
Resorcinol test
Normal fructose
Importance of evaluation
Count the number of dead cells
in 100 sperm
Brightfield / phase-contrast
 Living cells – blue
 Dead cells – red
Normal vitality: 50% or more
living cells correspond to the
previously evaluated motility.
Defective flagellum
Epididymal pathology
Low fructose level
(+) fructose = orange color
Equal to or greater than 13 μmol
per ejaculate
Spectrophotometric method
Tested within 2 hours
 Frozen to prevent fructolysis
Antisperm Antibodies Possible cause of infertility Occur following surgery,
Male / Female vasectomy reversal
(vasovasostomy), trauma, and
infection
 Clumps of sperm are Examination
observed
 Sperm-agglutinating
antibodies = sperm stick
to each other in a head-
to-head, head-to-tail, or
tail to- tail pattern.
 Graded as few,
moderate, or many on
microscopic
examination.
a. (MAR) test - screening Tests
procedure, detect the
presence of IgG antibodies
>10% = normal
b. Immunobead test – more
specific, detect IgG, IgM,
IgA antibodies
<50% = normal
Microbial and Chemical More than 1 million leukocytes Infection within the
Testing per millimeter reproductive system, frequently
the prostate
Test Chlamydia trachomatis, Microbial
Mycoplasma hominis, and
Ureaplasma urealyticum
Decreased neutral a - Suggest a disorder of the
glucosidase, epididymis.
glycerophosphocholine,
and L-carnitine
Decreased zinc, citric acid, Indicate a lack of prostatic fluid
glutamyl transpeptidase, and
acid phosphatase

Spectrophotometric methods Used to quantitate citric acid

and zinc
Rape evaluation  Enhanced with xylene and
examining under phase
microscopy
 Motile sperm can be
detected for up to 24
hours after intercourse
 Nonmotile sperm persist
for 3 days.
 As the sperm die off,
only the heads are
 present for 7 days after
intercourse
 High concentration of
prostatic acid phosphatase

Detection of seminal Specific method

glycoprotein p30
(prostatic specific antigen /
PSA), which is present even in
the absence of sperm.
Postvasectomy Semen Beginning at 2 months (+) / (-) results only
Analysis postvasectomy, continuing until
two consecutive monthly
specimens show no
spermatozoa.
Sperm Function Tests Performed in specialized Assess not only the
andrology laboratories characteristics of sperm but also
 include the hamster egg the functional ability
penetration assay,
cervical mucus
penetration test, hypo-
osmotic swelling test,
and the in vitro
acrosome reaction
Semen Analysis Quality The standardized procedures
Control developed by the WHO has
provided a basis for laboratory
testing and reporting.

The use of CASA has aided in

reducing the subjectivity of
the analysis. However, even
computerized analysis has been
shown to vary among operators

Summary