SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY

AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY

by the total number of sperm ejaculated (sperm
FULL NAME (LAST NAME, FIRST NAME): concentration x sperm volume)
AGUAYO, JOSHUA ARVIN A.
_______________________________________________________________
• Time since last sexual activity - Absence of ejaculation

makes the spermatozoa accumulate in the epididymis
and are soon flushed in the urine. However sperm vitality
“I hereby promise, as a Future Bedan Physician, that I will never cheat in this
exam, nor cheat in behalf of other people. I would not transmit/share the
is not affected by increased length of abstinence
content of this exam to anyone else. I am aware and I fully accept, any and all • Testicular Size. Size of the testes reflects spermatogenic
disciplinary sanctions that may be given to me if I fail to comply. activity as well as sperm morphology.

I am a Future Bedan Physician. I will become a doctor one day. I will answer EVALUATIVE QUESTIONS
this exam with honesty and integrity.”

ge
1. What are the structures involved in the production of
______________________________________________________________
semen? Explain the process of semen production until
Signature
ejaculation.

SEMEN ANALYSIS Seminiferous
↓ tubules -

for the
process of spermatogenesis

Learning Objective 2. Epididymis -
for maturation of sperm.

1. To be able to understand the structures involved in
sperm production and describe the semen components
3. Was Deferens -

propel sperm the

to
ejaculatory duct.

with regards to source and function 4. Seminal vesicles provides the
sperm with fluids nutrients
-

and

2. Describe the flow of semen from production to high in fructose.

ejaculation
3. To be able to describe, explain and understand the 5. Prostate Gland-has enzymes and proteins for coagulation and

different components of semen analysis liquefaction.


You may use the following references:
C. Bulborethral Gland-produces alkaline mucus thataids

WHO 2010 Lab Manual for the Examination and Processing of Human Semen
in neutralizing ofthe
the acidity
Fifth Edition. Publication of the World Health Organization. Geneva vagina's
Switzerland. 2010. secretory product.

males, testosterone andother hormones
Strasinger, SK. Di Lorenzo, MS. Urinalysis and Body Fluids Fifth edition. USA.
=D
During of
puberty
2008 transform into
will cause these cells to sperm cells. These

cells will undergo meiosis and moves the epididymis for maturation.

to

INTRODUCTION
SAMPLE COLLECTION


Semen is produced from a concentrated suspension of
Ideally, samples are collected in a room near the
spermatozoa composed of four fractious contributed by the
laboratory to limit exposure to temperature fluctuations and
following male reproductive organs : epididymis, seminal vesicles,
control the time from collection to analysis. Males should take
prostate and bulbourethral glands.
sexual abstinence at about 2-3 days, maximum of 5 days. Fertility

testing make use of 2-3 samples, collected at a 2 week interval
having two abnormal samples considered to be significant.
Specimens must be collected by masturbation.

1. In cases where collection of sample is done at home, state
the necessary steps from sample collection to transport at
the laboratory.
Give clear and written instruction for collection and
Strasinger, SK. Di Lorenzo, MS. Urinalysis and

-

transport.
Body Fluids Fifth edition. USA. 2008
patient should be given a pre-weighed container, labelled

-

The semen has two major quantifiable attributes with his name and ID number.

patientshould record the time of some production and
-

• Total number of spermatozoa reflecting sperm

deliver sample to the lab within 1 hour.
production and patency of post-testicular duct system

• Total fluid volume reflecting the secretory activity of -During transport, sample should be keptbly 20-37°C.
the accessory glands Reportshould note thatsample was collected from home/other
-

2. In cases where males demonstrate inability to produce location.

According to the WHO Laboratory Manual for sample by masturbation, how is semen collected using
examination and processing of human sperm, laboratory condoms?
measurements of sperm quality depends on:
Use special non-toxic condoms designed for semen collection;these
-

• Collection of a complete sample – First semen fractions

are also commercially available.
during ejaculations contains sperm-rich fluids. Latter
The patientshould be
fractions are composed of seminal vesicular fluids given info. from the manufacturer on how
- to

• Activity of the Accessory Glands – Fluids produced

use the condum, close it, and transportto lab.
dilute the concentration during ejaculation. Sperm
-Record the time of semen production and deliver simple the 19b to
concentration is not a direct measure of sperm count but
within 1 hour of collection.

During transport, sample should be keptb/w 20-37°C.
-

SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY

was collected
Page 1 of 3
Report
should note that sample by
-

AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY

means of special condum
during sexual intercase of home or other

location.
 SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY
AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY
STANDARD PROCEDURES IN SEMEN ANALYSIS 2. List three different methods on how semen volume is
measured. Briefly differentiate the methods used.
According to the WHO Laboratory Manual for What cause low semen volume? High semen volume?
examination and processing of human sperm, analysis of seminal compensatory vol. loss
semen volume can be measuredby:
fluid involves the following steps ↑
-Graduated
cylinder Final vol- observedvol. +0.5mL
-

First 5 minutes 10cc

syringle who the needle-aspirate completely wo air bubbles
~

• Placing specimen container in an incubator for in

slanting position. Add 0.1.

liquefaction Analytical weighing scale-1g.=ImL

-

Between 30 and 60 minutes
Low Vol Semen-Hypospermia
• Assessing liquefaction and semen appearance obstruction in ejaculatory duct.
-

• Semen volume measurement

congenital bilateral absence was deferens.of

• Semen pH assessment collection problems

·

• Wet smear to assess microscopic appearance, sperm partial retrograde ejoculation

motility, and dilution for sperm number androgen deficiency

-

• Semen smear to assess sperm morphology

High Vol. Semen- Hyperspennia
• Semen dilution to assess sperm concentration
-extended period ofs exual arousal/stimulation
• Mixed anti-globulin reaction (MAR) test (if required)
abstinence from sex and mosturbation

-

• Assessing peroxidase-positive cells (if round cells are -active exudation inflammation of
accessory glands
->

present)
3. Describe the normal appearance of semen
• Immunobead test (if required)

• Centrifuging semen (if biochemical markers are to be Normal: Abnormal:

assayed) whiteturbidity presence ofWBC
to Grayish white
-

Color:White
infection

-

Within 3 hours Appearance:Translucent Red coloration RB2

-

• Sending samples to the microbiology laboratory (if odor:musty/Bleach yellow coloration nine contamination

-

required) prolonged abstinence

-

or medication.

After 4 hours

• Fixing, staining and assessing smears for sperm

morphology





INITIAL MACROSCOPIC EXAMINATION



4. Normal pH of semen is alkaline. What conditions
causes increased pH? decreased pH? How is the pH
balance of the semen achieved?

Increased pH (8.0 infection
- =

Decreased pH <7.2=congenital bilateral absence of was
-

deferens/obstruction ofejaculatory
duct/

pH balance of is achieved
-
the semen through the

basic seminal vesicle secutions and acidic

Kindly watch this laboratory guide before answering the prostatic secretions.
Evaluative questions

1. Explain the process of liquefaction. What are the
causes of liquefaction failure? 5. Discuss Semen Hyperviscosity. What are its
implications on fertility?
-

Semen is typically a somi-solidcoagulatedmass. Within

Semen is
Hyperviscosity abnormally viscous
~

an

a few minutes, semen will begin to liquery (thinner),

neterogenous mixture
of lumps will be seen. As it continues,
semen
exhibiting homogeneous stickiness
a

and consistency caused by malfunctions in

the semen will become more homogeneous and quite
the
seminal reside and prostate.

watery. This happens up to60 mins upon collection.
This could impair sperm motility and
infertility.

Delayed liquefaction may indicate
deficiency in prostotic enzymes
-

or infection.



SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY
Page 2 of 3
AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY
 SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY
AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY
b. What tests could be performed to further
INITIAL MICROSCOPIC EXAMINATION evaluate the patient’s infertility

2 tests for decreased motility and clumping:
Reaction test

-

Mixed Agglutination
-
Immunobead test



Sperm Motility Sperm Viability
Reaction test

·
Mixed Agglutination
-used primarily detect
presence of IgG.
for
screening to
-

awarearemaine
Yen

c. Briefly explain the different interpretations
offered by these two tests


Sperm Morphology Sperm Count

<10% ofthe motile sperm attached:Normal
-

Kindly watch this laboratory guide before answering the

Immuno bead test
Evaluative questions
·

more
specific procedure used presence ofIgG,
detect
to

1. Explain the grading of sperm motility. Igm, determines whatarea do these antibodies
IgAand
of sper
Forward and progressive movement affect.

sperm are mixed w/ polyacrylamide beads to wi

be coated

-

either anti-IgG, anti-Igm, antilgA and these beads

Grade attached the sperm be microscopically seen going
to can

4 (a)- rapid, straight-line motility particular
to areas.

presence of beads <50% is considered normal WHO.

-

3 (b) -

slower speed, some lateral movement

d. State three ways in which a positive result on
later
2 (b) -

slow forward progression, noticeable these tests could be affecting male fertility
movement affects
Clumping the quality amountof sperm.
-

and

I (c) -
No forward progression
will affect

-

Orum penetration change in

morphology
⑧ (d) - No movement the
way sperm penetrates ovum

-motility
focuses on
sperm penetration.
2. A yellow colored semen specimen is received in the

laboratory. The analysis is normal except for

decreased sperm motility. Explain the possible

connection between these two findings


colored
yellow semen urine
could indicate contamination,
-

prolonged abstinence, and even medications (VitB, antibiotics for R3).



-

Urine is toxic to the sperm decreasing motility.

3. A semen analysis on a post-vasectomy patient has
normal sperm concentration but motility is decreased
and clumping is observed on wet preparation
a. Explain the possible connection between
these observations and the patient’s recent
surgery.

Presence of anti-sperm antibodies. Clumps are observed

-

This happens when the

during routine semen analysis.
blood-testis barrier is disrupted usually caused
by
infections. Antigens
vasectory, operations, traud or

on
sperm may trigger
the immune an response damaging
of
sperm cells. Damaged sperms may
the cause production

antibodies in the female.

SAN BEDA UNIVERSITY COLLEGE OF MEDICINE – SECTION OF PHYSIOLOGY

Page 3 of 3
AY 2021-2022 REPRODUCTIVE PHYSIOLOGY LABORATORY