SEMENALYSIS OR SEMINAL FLUID ANALYSIS

• Semenalysis or seminal fluid analysis is the analysis or examination of seminal fluid.
• Spermatozoa accounts for about 5% of the fluid and comes from the testes and epididymis.
• ACP activity is conveniently detected in a spot test in which hydrolysis of substrate is coupled to a color reaction (Brentamine test); a test is scored as positive when the color appears within a defined time frame.
• Microscopic examination of sperm cells is used to identify ACP activity.
• Fluid from the seminal vesicle accounts for about 60 - 70% of the fluid.
• Fluid from the prostate glands accounts for about 20 - 30% of the fluid.
• Fluid from the bulbourethral glands accounts for about 5% of the fluid.
• Semen plasma is the nutritive medium in the fluid and aids in the fertilization process.
• Choline and spermine keep the sterility of seminal fluid.
• Fructose is the main sugar of ejaculate and a major nutrient of spermatozoa.
• Proteolytic enzyme (fibrinolysin) is responsible for coagulation and liquefaction.
• Carbonates contribute to the alkalinity of the seminal fluid.
• Acid phosphatase and lactate dehydrogenase are secreted in the prostate glands and present in human seminal fluid, absent in other mammals.
• Normal seminal fluid is usually grayish white and translucent, while abnormal seminal fluid may have increased white turbidity, reddish, or yellowish color.
• Normal seminal fluid should liquefy after 30 - 60 minutes, while abnormal seminal fluid may fail to liquefy due to low prostatic enzyme secretion of the prostate.
• Normal seminal fluid volume is 2 - 5 ml/ ejaculate, while increased seminal fluid volume may be associated with extended/prolonged abstinence or decreased seminal fluid volume may be associated with infertility.
• Normal seminal fluid odor is Fishy, distinct musty or chlorox-like, while abnormal seminal fluid odor may be different.
• Normal seminal fluid pH is 7.2 to 8.0, while abnormal seminal fluid pH greater than 8.0 may indicate bacterial infection within the reproductive tract or less than 7 may be due to problem with prostatic secretions and bulbourethral secretions.
• Methods of determining sperm motility include manual and computer assisted semen analysis (CASA).
• Revitalization test uses 1 drop of glucose ringer’s solution to differentiate true dead from immobilized sperm cells.
• Bloom’s Eosin test is a follow-up test if sperm count is normal but motility is decreased.
• HYPO-OSMOTIC SWELLING involves exposing sperms to low sodium concentration and evaluating the membrane integrity and viability.
• Antisperm antibodies (ASA) are immunoglobulins IgG, IgA, and/or IgM that impair sperm functions.
• IN-VITRO ACROSOME REACTION involves evaluating the acrosome to produce enzyme for ovum penetration.
• FLORENCE TEST is based on the presence of spermine crystals to determine choline present in semen; it only indicates presence of semen.
• Condensation with indole acetic acid principle: fructose from semen is treated with concentrated HCl to convert fructose to oxymethyl furfural state which later condenses with indole-3-acetic acid forming a purple colored compound and is measured spectrophotometrically.
• Barbiero’s Test involves using reagents such as 2.5% TCA, saturated picric acid solution.
• CERVICAL MUCUS PENETRATION involves allowing sperms to react with a mid-cycle cervical mucus.
• Produces an orange color when fructose is present.
• Immunobead test is a more specific procedure that can detect presence of IgA, IgG and IgM antibodies and can demonstrate what area of sperm cell the autoantibodies are affecting.
• Sperms are incubated with species-nonspecific hamster eggs and penetration is observed microscopically in Hamster Egg Penetration test.
• FLUORESCENCE/ UV test involves dried sperm cells giving a brilliant pale lavender fluorescence under UV light or wood’s lamp.
• Photometric somogyi filtration principle: uses resorcinol for condensation.
• Anti-sperm antibodies (ASA) are immunoglobulins IgG, IgA, and/or IgM that impair sperm functions.
• Sperms are evaluated for the production of enzyme for ovum penetration in In-vitro Acrosome Reaction test.
• Methods of determination for antisperm antibodies include Mixed Agglutination Reaction (MAR) and Immunobead test.
• If antisperm antibodies are present in a sample, fertility treatment can help.
• Reagents: TCA; indole-3-acetic acid; concentrated HCl.
• Florence Test is based on the presence of spermine crystals to determine choline present in semen; it only indicates presence of semen.
• Fluorescence/ UV test involves dried sperm cells giving a brilliant pale lavender fluorescence under UV light or wood’s lamp.