SEMEN ANALYSIS FOR INVESTIGATION OF INFERTILITY
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Semen (or seminal fluid) is a fluid that is emitted from the male genital tract and contains sperms that are capable of fertilizing female ova. Structures involved in production of semen are:
- Testes: Male gametes or spermatozoa (sperms) are produced by testes; constitute 2-5% of semen volume.
- Epididymis: After emerging from the testes, sperms are stored in the epididymis where they mature; potassium, sodium, and glycerylphosphorylcholine (an energy source for sperms) are secreted by epididymis.
- Vas deferens: Sperms travel through the vas deferens to the ampulla which is another storage area. Ampulla secretes ergothioneine (a yellowish fluid that reduces chemicals) and fructose (source of nutrition for sperms).
- Seminal vesicles: During ejaculation, nutritive and lubricating fluids secreted by seminal vesicles and prostate are added. Fluid secreted by seminal vesicles consists of fructose (energy source for sperms), amino acids, citric acid, phosphorous, potassium, and prostaglandins. Seminal vesicles contribute 50% to semen volume.
- Prostate: Prostatic secretions comprise about 40% of semen volume and consist of citric acid, acid phosphatase, calcium, sodium, zinc, potassium, proteolytic enzymes, and fibrolysin.
- Bulbourethral glands of Cowper secrete mucus.
|• Testes and epididymis: 10%
• Seminal vesicles: 50%
• Prostate: 40%
• Cowper’s glands: Small volume
Normal values for semen analysis are shown in Tables 835.1 and 835.2.
|1. Volume||≥2 ml|
|2. pH||7.2 to 8.0|
|3. Sperm concentration||≥20 million/ml|
|4. Total sperm count per ejaculate||≥40 million|
|5. Morphology||≥30% sperms with normal morphology|
|6. Vitality||≥75% live|
|7. White blood cells||<1 million/ml|
|8. Motility within 1 hour of ejaculation|
|• Class A||≥25% rapidly progressive|
|• Class A and B||≥50% progressive|
|9. Mixed antiglobuiln reaction (MAR) test||<50% motile sperms with adherent particles|
|10. Immunobead test||<50% motile sperms with adherent particles|
|1. Total fructose (seminal vesicle marker)||≥13 μmol/ejaculate|
|2. Total zinc (Prostate marker)||≥2.4 μmol/ejaculate|
|3. Total acid phosphatase (Prostate marker)||≥200U/ejaculate|
|4. Total citric acid (Prostate marker)||≥52 μmol/ejaculate|
|5. α-glucosidase (Epididymis marker)||≥20 mU/ejaculate|
|6. Carnitine (Epididymis marker)||0.8-2.9 μmol/ejaculate|
INDICATIONS FOR SEMEN ANALYSIS
|• Physical examination: Time to liquefaction, viscosity, volume, pH, color
• Microscopic examination: Sperm count, vitality, motility, morphology, and proportion of white cells
• Immunologic analysis: Antisperm antibodies (SpermMAR test, Immunobead test)
• Bacteriologic analysis: Detection of infection
• Biochemical analysis: Fructose, zinc, acid phosphatase, carnitine.
• Sperm function tests: Postcoital test, cervical mucus penetration test, Hamster egg penetration assay, hypoosmotic swelling of flagella, and computer-assisted semen analysis
Availability of semen for examination allows direct examination of male germ cells that is not possible with female germ cells. Semen analysis requires skill and should preferably be done in a specialized andrology laboratory.
- Investigation of infertility: Semen analysis is the first step in the investigation of infertility. About 30% cases of infertility are due to problem with males.
- To check the effectiveness of vasectomy by confirming absence of sperm.
- To support or disprove a denial of paternity on the grounds of sterility.
- To examine vaginal secretions or clothing stains for the presence of semen in medicolegal cases.
- For selection of donors for artificial insemination.
- For selection of assisted reproductive technology, e.g. in vitro fertilization, gamete intrafallopian transfer technique.
COLLECTION OF SEMEN FOR INVESTIGATION OF INFERTILITY
Semen specimen is collected after about 3 days of sexual abstinence. Longer period of abstinence reduces motility of sperms. If the period of abstinence is shorter than 3 days, sperm count is lower. The sample is obtained by masturbation, collected in a clean, dry, sterile, and leakproof wide-mouthed plastic container, and brought to the laboratory within 1 hour of collection. The entire ejaculate is collected, as the first portion is the most concentrated and contains the highest number of sperms. During transport to the laboratory, the specimen should be kept as close to body temperature as possible (i.e. by carrying it in an inside pocket). Ideally, the specimen should be obtained near the testing site in an adjoining room. Condom collection is not recommended as it contains spermicidal agent. Ejaculation after coitus interruptus leads to the loss of the first portion of the ejaculate that is most concentrated; therefore this method should not be used for collection. Two semen specimens should be examined that are collected 2-3 weeks apart; if results are significantly different additional samples are required.
• Microscopic examination for (i) percentage of motile spermatozoa, (ii) sperm count, and (iii) sperm morphology
EXAMINATION OF SEMINAL FLUID
The tests that can be done on seminal fluid are shown in Box 835.2. Tests commonly done in infertility are shown in Box 835.3. The usual analysis consists of measurement of semen volume, sperm count, sperm motility, and sperm morphology.
Terminology in semen analysis is shown in Box 835.4.
EXAMINATION OF SEMEN TO CHECK THE EFFECTIVENESS OF VASECTOMY
|• Normozoospermia: All semen parameters normal
• Oligozoospermia: Sperm concentration <20 million/ml (mild to moderate: 5-20 million/ml; severe: <5 million/ml)
• Azoospermia: Absence of sperms in seminal fluid
• Aspermia: Absence of ejaculate
• Asthenozoospermia: Reduced sperm motility; <50% of sperms showing class (a) and class (b) type of motility OR <25% sperms showing class (a) type of motility.
• Teratozoospermia: Spermatozoa with reduced proportion of normal morphology (or increased proportion of abnormal forms)
• Leukocytospermia: >1 million white blood cells/ml of semen
• Oligoasthenoteratozoospermia: All sperm variables are abnormal
• Necrozoospermia: All sperms are non-motile or non-viable
The aim of post-vasectomy semen analysis is to detect the presence or absence of spermatozoa. The routine follow-up consists of semen analysis starting 12 weeks (or 15 ejaculations) after surgery. If two successive semen samples are negative for sperms, the semen is considered as free of sperm. A follow-up semen examination at 6 months is advocated by some to rule out spontaneous reconnection.
- PHYSICAL EXAMINATION OF SEMEN FOR INVESTIGATION OF INFERTILITY
- BIOCHEMICAL ANALYSIS OF SEMEN FOR INVESTIGATION OF INFERTILITY
- MICROSCOPIC EXAMINATION OF SEMEN FOR INVESTIGATION OF INFERTILITY
- IMMUNOLOGIC ANALYSIS OF SEMEN FOR INVESTIGATION OF INFERTILITY
- SPERM FUNCTION TESTS OR FUNCTIONAL ASSAYS
- EXAMINATION FOR THE PRESENCE OF SEMEN IN MEDICOLEGAL CASES