SEMEN EVALUATION

SEMEN EVALUATION
PRACTICAL VIEW
BY
Hassaan Ali Gad
Assistant lecturer of urology and Andrology
Aswan University

OUTLINE
• What is semen
• Formation of the sperm cell
• WHY PERFORM SEMEN ANALYSIS
• What is the “standard” approach to semen
evaluation?
Hassaan Ali 20154/7/2015

What is semen
A mixture of seminal plasma and cells
• Seminal plasma contains:
– Prostatic fluid (~30% of the volume)
– Epididymal plasma (~5% of the volume)
– Seminal vesicle fluid (the remainder of the ejaculate)
• The cells are:
– Spermatozoa
– Germ line cells
– Leukocytes of various types
– Bacteria
– Epithelial cells
– Occasional red cells

Spermatogenesis is a cascade of cell divisions:
Mitosis: spermatogonia to
primary spermatocytes
First meiotic division:
secondary spermatocytes
Second meiotic division:
haploid spermatids
This process takes 70 ± 4 days
in the human – so errors will
take about 3 months to show
up

Spermiogenesis:
differentiation of the round spermatid into a spermatozoon

WHY PERFORM SEMEN ANALYSIS?
•Diagnosis of sterility
•Diagnosis of infertility
•Prognosis for fertility
•Identify treatment options:
• surgical treatment
• medical treatment
• assisted conception treatment

What is the “standard” approach to
semen evaluation?
•World Health Organization’s Lab Manual fifth edition in 2010..
•Focus is on standardization with expanded section on quality
control.
•Methods amenable for use in any (“third world”) country.
•Basic infertility work-up.

Sample Collection
• For a meaningful result, semen samples must always be
collected under standardized conditions:
– the container has to be sterile and known NOT to be
spermotoxic (i.e. provided by the lab)
– the man must have had 3 – 5 days of abstinence
– the man must have washed his hands before collection
(particularly if microbiological analysis is requested)
– the sample must be kept at 37°C until analysis, which begins
ideally within 30 min, but absolutely within 60 min, of
ejaculation

Methods Collection
1. Masturbation (the method of choice for all seminal fluid
tests).
2. By condom: it is not recommended for fertility testing
because the condoms may contain spermicidal agents.
3. By coitus interrupts: (withdrawal method).
4. Percutaneous epididymal sperm aspiration (PESA).
if a blockage in the vas deferens is suspected, semen can be
taken directly from the epididymis
5. TESA: Testicular sperm extraction (TESE)- Open Testicular
Biopsy: is a highly invasive, open surgical procedure
performed under general anaesthetic. The scrotum and
testes are cut open, before testicular tissues are cut away
and examined for sperm, which, if present can be extracted.

Semen Study
– There are several macroscopic and microscopic evaluations
which give useful diagnostic information about the sample
–Macroscopic microscopic
– Appearance
– Odour
– Liquefaction
– Volume
– Viscosity
– pH
-concentration/count
- motility
-morphology
-viability

WHO 2010
Lower Reference LimitParameter
1.5Semen volume (ml)
15Sperm concentration (106/ml)
39Total sperm number
(106/ejaculate)
32Progressive motility (PR, %)
40Total motility (PR +NP, %)
58Vitality (live sperms, %)
4Sperm morphology (NF, %)
>/=7.2pH*
<1Leucocyte* (106/ml)
<50MAR/Immunobead test* (%) Hassaan Ali 20154/7/2015

Nomenclature for semen variable
(WHO)
• Normal semen quality Normospermic
• No ejaculate Aspermia
• Low volume Hypospermia
• High volume Hyperspermia
• No spermatozoa Azoospermia

Nomenclature for semen variable
(WHO)
• Low spermatozoa conc. Oligozoospermia
• Low spermatozoal motility Asthenozoospermia
• Low normal morphology Teratozoospermia
• Excessively high sperm conc., Polyzoospermia
• WBCs in semen Pyospermia
• RBCs in semen Heamatospermia
• Non viable sperms (dead). Necrozospermia

Factors affecting SA results
• Medicines senomroh elamef dna elam ,enidtiemic sa hcus ,
dna ,niotnarufortin ,enizalasaflus ,)negortse,enoretsotset(
senicidem yparehtomehc emos.
• Caffeine occabot gnikoms dna ,anaujiram ,eniacoc ,lohocla ,.
• Herbal fo sesod hgih dna trow s'nhoJ .tS sa hcus ,senicidem
aecanihce.
• Temperature -fi wol yletaruccani eb lliw eulav ytiltiom mreps
dloc steg elpmas nemes eht.
• Exposure to radiation niatrec sa hcus( slacimehc emos ,
erusopxe taeh degnolorp dna ,)sedicimreps ro sedictisep.
• An incomplete semen sample --si elpmas a fi nommoc erom
notiabrutsam naht rehto sdohtem yb detcelloc.
• Not ejaculating for several days -- affect the semen volume

Macroscopic
• Volume
• Normal:1.5 ml per ejaculation
• Low volume (<1ml) reflect a problem with the seminal vesicles and
prostate – a block, retrograde ejaculation, infection or lack of
androgen.
• Low semen volume cannot neutralize vaginal acidity
• High semen volume dilute sperms/ active infection
• pH
• Normal:=/>7.2 (alkaline)
• Increased pH is indicative of infection within the reproductive tract.
• A decreased pH Seminal vesicle dysfunction or agenesis.
• Vas deferens agenesis.
• Ejaculatory ducts obstruction.
• Incomplete collection.
• .

Macroscopic
• Appearance
• Normal:Whitish to grey opalescent
• Yellow (urine, jaundice);
• Pink/Reddish/Brown (RBCs)
• Liquefaction
• Normal:15–30 minutes after collection
• Lumpy >60 min – sperms may be trapped in unliquefied jelly; maybe
sign of prostatic infection, lack of prostatic protease
• Viscosity
• Normal;Smooth and watery
• Abnormal –, thick with long threads (21G needle). High viscosity
impede sperm movements

sperm count
• Certain conditions may be linked with a low or absent
sperm count. These conditions include :
• Orchitis
• Varicocele
• Klinefelter syndrome
• Radiation treatment to the testicles
• Diseases that can cause shrinking (atrophy) of the testicles
(such as mumps).
• Long term illness such as diabetes which may cause
retrograde ejaculation
• Hormonal imbalance (testosterone, luteinizing hormone
(LH), follicle-stimulating hormone (FSH), or prolactin
• Azoospermia - A small sample (biopsy) of the testicles may
be needed for further evaluation.

Motility
• Fertilization of an ova is dependent on the ability of the sperm to reach
and unite with it.
• Grade 4 (A): Sperm with progressive motility. These are the strongest and
swim fast in a straight line.
• Grade 3 (B): Sperm with non-linear forward motility. These also move
forward but tend to travel in a curved or crooked motion.
• Grade 2 (C): These have non-progressive motility because they do not
move forward despite they move their tails .
• Grade 1 (D): These are immotile and fail to move at all .

Morphology
• Refers to the shape and size of the sperm. A normal
sperm has an oval head and a tail 7- 15 times longer
than the head. Abnormal sperm morphology can be a
contributing factor to infertility.
• Causes of abnormal morphology include;
• Congenital testicle abnormalities.
• Fever.
• If any abnormal morphology is seen the semen analysis
should be repeated in four to two weeks to check if the
changes are temporary or permanent.
.

Vitality
• Distinguishes between samples with live, but
immotile sperm (e.g Kartagener syndrome),
and those with lots of dead sperm (could
result from: sperm senescence;
• exposure to detergents or lubricants; or
spermotoxic antibodies)

Other cells in semen
• Leukocytes: normally (1-4/HPF), increase number
(leukocytospermia) indicates reproductive tract infection
• Epithelial cells: normally (1-2/HPF)
• Spermatocytes: (Immature germ cells) 1-2/HPF.
• Erythrocytes: (1-2/HPF). Increased number may indicate a
reproductive tract infection or damage to a small capillary
during sample production.
• Note: bacteria and protozoan such as Trichomonas vaginalis
are uncommon in human semen but their presence is
indicative of possible male reproductive tract infection and
should be reported to the referring doctor for further
evaluation.

Semen biochemistry
• Acid phosphatase: marker for prostatic function
• Citric acid: can indicate prostatic function – low levels may
indicate dysfunction or a prostatic duct obstruction
• Zinc: marker for prostatic function – colorimetric assay
(WHO)
• Fructose: marker for seminal vesicle function, and is a
substrate for sperm metabolism – spectrophotometric
assay (WHO)
• -Glucosidase: secreted exclusively by the epididymis and
so is a marker for epididymal function – spectrophotometric
assay (WHO)

• A CASA system is an automated, objective and standardized
equipment that allows to assess all the parameters of
sperm in a semen sample.
• CASA are most often used for the assessment of sperm
concentration and motility following the WHO criteria.
• By use of CASA several specific motility parameters in a
more detailed manner such as velocity can be obtained.

Advantages of CASA
• Advantages
– More objective and reproducible measurement
– Superior documentation of laboratory values
– Superior in measurement of sperm motility
• Disadvantages
– Not reliable if sperm density is <2x106/ml or >50x106/ml, lots of
debris/immotile sperm. Require dilution if >40 x 106/ml in
isotonic buffer to avoid sperm collision
– Need to record 200 sperms for accurate distribution of velocity.
– Parameters not standardized between laboratories –difficult to
interpret results
– No improvement on the manual method in distinguishing
fertilizing capacity of semen

SEMEN EVALUATION

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