AUBF - Semen Analysis

Azoospermia

Absence of Spermatozoa in Semen or Seminal Fluid.

Aspermia

No semen.

Spermatid

Immature/Young Sperm Cells.

Necrospermia

Presence of dead sperm cells.

Oligospermia

Decreased number of sperm.

Asthenozoospermia

Low percentage of progressively motile spermatozoa/sperm cells.

Or may imply a high percentage of either:

• Non-progressively motile sperm (e.g., moving in circles or twitching)

• Non-motile sperm (no movement at all)

• Or both

Teratozoospermia

Low percentage of morphologically normal sperm.

C. Both are True

Seminal Fluid or Semen is a composite solution formed by testes as well as other male reproductive organs.

It consists basically of spermatozoa suspended in seminal plasma.

a. First Statement is True. Second Statement is False.

b. First Statement is False. Second Statement is True.

c. Both are true.

d. Both are false.

Testis → Epididymis → Vas Deferens → Seminal Vesicle → Ejaculatory Duct → Prostate Gland → Bulbourethral Gland → Urethra

Flow of Semen

Testes

Responsible for producing sperm cells, and secreting testosterone. It is also the site of spermatogenesis in the seminiferous tubules. Most important component of the seminal fluid.

Epididymis

It is the site of maturation of sperm, where sperm cells become motile.

Vas Deferens

This propels sperm to the ejaculatory duct.

Seminal Vesicle

A convoluted, pouch-like, that also acts as the transport medium for the sperm and produces an alkaline (7.2 - 8 pH) viscous fluid. This also contains the seminal plasma. This provides nutrients for sperm (fructose) and also contains clotting proteins.

Prostate Gland

A Golf-Ball-Like that produces a slightly acidic fluid (6.5). It contains enzymes that can break down clotting proteins, liquefying semen.

Bulbourethral Gland or Cowper’s Gland

Pea-size part of the male reproductive system that produces a thick alkaline fluid to neutralize the vaginal acidity present in vaginal flora and protects the passing of sperm by neutralizing the acid from urine and prostate gland in the urethra. It also secretes mucus that lubricates the penis and the lining of the urethra.

Testes - 5%

Seminal Vesicle - 60% to 70%

Prostate Gland - 20% to 30%

Bulbourethral Gland - 5%

Four Fractions of Seminal Fluid

Sperm Cells and Prostatic Fluid.

Components of the First Portion of the Ejaculate.

Mostly Seminal Vesicle Fluid

Components of the Second/Last Portion of the Ejaculate.

Sperm Count - Decreased

pH - Falsely Increased (Alkaline)

Specimen - Will not liquefy

Effects of Missed First Portion of the Ejaculate.

Semen Volume - Decreased

Sperm Count - Falsely Increased

pH - Falsely Decreased (Acidic)

Specimen - Will Not Clot

Effects of Missed Last Portion of the Ejaculate.

Seminal Vesicle - Alkaline

Prostate Gland - Acidic

Bulbourethral Gland - Alkaline

pH of Seminal Vesicle, Prostate Gland, and Bulbourethral Gland

Varicocele

This is the most common cause of infertility for male. It is formed when valves inside the veins along the spermatic cord prevent blood from flowing properly, causing the swelling and widening of the veins (hardening of veins that drain the testes). And one of the signs in semen analysis for this is the tapering of the head of the sperm cells.

Mumps or Beke

This one cause infertility around 40% of the time/cases.

1. Varicocele

2. Mumps

3. Klinefelter’s Syndrome (XXY)

4. Malignancy

4 Causes of Infertility

Spermatogenesis

This term means the formation and development of sperm in the seminiferous tubules of the testes.

Spermiogenesis

This term refers to the maturation of spermatids into sperm cells.

90 Days

How long does spermatogenesis take?

Start of Spermatogenesis

Spermatogonium → Mitosis = 1 Spermatogonium , 1 Primary Spermatocyte or Sperm Cells

Primary Spermatocyte → Meiosis I = Secondary Spermatocyte

Secondary Spermatocyte → Meiosis II = Spermatid

Spermatid → Spermiogenesis = Sperm Cell

End of Spermatogenesis

Flow of Sperm Cell Maturation

Sertoli Cells

Cells responsible for providing support and nutrients for the germ cells (spermatogonium, spermatocytes) as they undergo mitosis and meiosis.

Ejaculatory Ducts

This receives both the sperm from the ductus deferens and fluid from the seminal vesicles.

Fructose, Prostaglandin, Clotting Proteins, and Flavin

Fast Forward Po, Clotting Proteins

Components of Seminal Vesicle

Fructose

Semen component that provides nutrients to the sperm cells.

Prostaglandin

Component of semen responsible for sperm motility and viability.

Clotting Proteins

Component of semen responsible for coagulation of semen after about 5 seconds of ejaculation.

Flavin

Component of semen that makes its appearance Gray or Opalescent, as well as the Blue to Yellow Fluorescence when semen is visualized under ultraviolet light (Wood’s Lamp) or Geen-White Fluorescence under UV Light.

Proteolytic Enzymes

Prostatic Specific Antigen

Citric Acid

Acid Phosphatase

Choline and Spermine

Zinc

PePsa, CaAp, Choline and Spermine, Zinc

Components of the Prostatic Fluid

Citric Acid

Component of semen used by sperm for ATP production.

Proteolytic Enzymes

Component of semen that breaks down clotting proteins and liquefy semen.

Usually, 30 to 60 minutes after collection.

Liquefaction Time

Choline and Spermine

Component of semen that inhibit bacterial growth.

1. Masturbation

2. After Interrupted Sexual Intercourse [Coitus Interrupts or Withdrawal Method]

3. Condom

4. Aspiration from Vaginal Canal after Intercourse

Collection Methods

St. Lukes Medical Center

For IVF testing, other than doing it in U.S., Japan, or Singapore, what hospital here in the Philippines offer IVF testing already?

After Interrupted Sexual Intercourse [Coitus Interrupts or Withdrawal Method]

Collection method that should be used when a patient is not allowed to masturbate because it is against their beliefs.

Sperm Clumping

Sign of Anti-sperm Antibodies

1. Empty Bladder

2. Sexual Abstinence

3. Fertility Testing

4. Warm Sterile Glass

Requirements prior to collection of semen.

1. Ideally kept at room temperature or body temperature [37C]

2. Delivered to the laboratory within 1 Hour of Collection

Requirements for transport of semen.

Color: Grayish White or Pearly White

• Translucent

  • Not Clear, Not Turbid

    • Clear: No Sperm Cells

    • White Turbid: Many WBCs and Infection

Odor: Musty or Bleach-Like

Normal Appearance/Color or Odor of Semen.

Clear: Low Sperm Count

Yellow: Urine Contamination, Prolonged Abstinence, Medications

Gray: Flavin

Red: RBCs (Abnormal)

Colors of Semen (Clear, Yellow, Gray, and Red) and Its Meaning.

Deficiency in Prostatic Enzymes or Proteolytic Enzymes.

It should be reported.

Indication if the liquefaction time took more than 60 minutes.

After 2 Hours

After how many hours, before you initiate putting proteolytic enzymes in a specimen that took too long to liquefy?

1. Dulbecco's phosphate-buffered saline (DPBS), or

2. alpha-chymotrypsin bromelain

If after 2 hours the specimen has not liquefied, add _____ or proteolytic enzymes such as _____.

2 to 5 ml per ejaculate

Normal Volume of Semen

Pours like Droplets

Normal Viscosity of Semen

7.2 to 8

Normal pH of Semen

Infection

Indication of High Seminal pH

• Increased Prostatic Fluid

• Obstruction of the Ejaculatory Duct

• Problem in the Seminal Vesicle

Indication of Low Seminal pH

1. Sperm Concentration

2. Motility

3. Morphology Percentage

3 Microscopic Examination for Semen Analysis

20 to 250 Million Sperm PER ML

Sperm Concentration Reference Value

10 to 20 Million Sperm PER ML

Sperm Concentration Borderline Reference Value

AT LEAST 40 Million PER EJACULATE

Sperm Count Reference Value

Sodium bicarbonate

Diluting fluid that dissolves mucus.

Formalin

Diluting fluid that immobilizes sperm

Chilled Water [Not Tap]

Diluting fluid that we always use in the laboratory. The cheapest and always available

Saline and Distilled Water

Diluting fluid other than the sodium bicarbonate, formalin, or chilled water.

Sperm Conc. / mL = [# of Cells Counted x DF (20) x 1,000] / [# of Squares (2) x Vol. of 1 Square (0.1)]

or

Sperm Conc. / mL = # of sperm cells counted x 100,000

• if the dilution factor is 1:20 and the WBX squares are 2.

or

Sperm Conc. / mL = [# of Cells Counted x DF (20) x 1,000] / [# of Squares (5) x Vol. of 1 Square (0.004)]

or

Sperm Conc. / mL = # of sperm cells counted x 1,000,000

• if the dilution factor is 1:20 and you’re counting in 5 RBC Squares.

Sperm Conc. Formula

Sperm Count/Ejaculate = Sperm Conc. x Semen Volume

Sperm Count / Ejaculate Formula

Sperm Motility Grading [Grade - WHO Criteria - Sperm Motility]

Reference Values for Semen Analysis: Volume, Viscosity, pH, Concentration, Count, Motility, Quality of Motility, Morphology