Semen Analysis

Semen Analysis1Objective:Physiology of semen formationIndication of semen analysisMacroscopic examinationof semenMicroscopic examinationAntibody testing in semenReference ranges2 Physiology of Seminal fluid Semen is grey opalescent fluid , which consists of suspension of spermatozoa in seminal plasma.

It is made up of the secretions of all accessory glands of the male genital tract.

3The Percentage contribution of Each of the Secretions in Seminal Fluid% EjaculateSource of Secretion5Testis40-80Seminal vesicles13-33Prostate2-5Bulbo-uretheral & uretheral glands4

Fructose: imp energy for sperms especially needed for sperms motilityProstaglandins: play a role in controlling sperm movement & sperm penetration of cervical mucusFibrinogen-like substance : acted upon by enzyme vesiculase to induce clotting that occurs in semenThe prostatic fluid pH =6.5Enzymes: vesiculase

Acid phosphatase

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Bulbo-uretheral & uretheral glands

The fluid rich in mucoprotein , so it lubricates the urethra

Occasionally in the secretions contain antisperm antibodies( may be of importance in infertility cases)

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7During ejaculation , each of the components that make up semen may be discharged from the urethra in a sequence

The first part is made up of testicular component of semen followed by secretion of the prostate , lastly the secretion of the vesicles are expelled.

8Indication of Semen AnalysisAssessment of fertility/infertility ( most common)Determination the effectiveness of vasectomyDetermination of suitability of semen for artificial inseminationFollow up of fertility after cancer treatment by radio- or chemotherapyForensic purpose9Semen analysisSpecimens:2 samples should be collected for initial evaluation,

the interval between the two collections should not be less than 7 days or more than 3 weeks ,

if the results of these assessments are markedly different , additional samples should be examined

10Examination of SemenMacroscopic ExaminationVolumeLiquefaction timeAppearance pHViscosityMicroscopic ExaminationSperm countSperm motility Sperm viability Sperm morphologyAgglutinationAntibodies coating of sperms Biochemical TestsFructose testAcid phosphatase11Macroscopic Examination12Volume : ( 2-6ml/ ejaculation)

Aspermia: total absence of ejaculation ( rare)Hypospermia or oligospermia: the seminal fluid is < 2mlHyperspermia: volume > 10 ml

( spermia denotes seminal fluid not spermatozoa)1313Liquefaction time: ( forms gel-like clot immediately after ejaculation)a normal semen sample liquefies with 15 -60 mint . ( prolongation must be recorded).

14Appearance:

homogeneous , grey-opalescent appearance The color is due to high content of protein &presence of > 60 million sperms/mlMay be less opaque if sperm concentration is very lowDense white turbid in inflammation and high WBCsHaematospermia :when red blood cells are present (blood)Yellow in patient with jaundice (very bright yellow due to bilirubin) or taking some vitaminsOr contaminated with urine ( uriniferous odour)15pH : between7.2-8.0 recorded in fresh semen by using pH paper .A patient exceeding pH 8.0 may suggest acute disease of the seminal vesicles Lowering of pH may be due chronic inflammatory of seminal vesiclesOr contamination with urine

16Viscosity :

Normal viscosity is that which allows semen to be poured drop by drop out of the container. In case of abnormal viscosity drop will form a thread more than 2 cm long High viscosity : can interfere determination of sperm motility17Microscopical Examination18Sperm Count:

Total number of sperms in an ejaculation. It is 20 million/ml i.e. 60 million / ejacIt is obtained by multiplying the sperm concentration by the volumeAzoospermia: means no spermatocytes ( male sterility)Oligozoospermia: < 20 million/ml (less than 50 million/ejaculationPolyzoospermia : may reach 350millions/ejaculation19Sperm CountDecreased:vasectomyvaricoceleprimary testicular failure (Klinefelters)secondary testicular failurecongenital vas deferens obstructionendocrine causes (prolactinemia, low testosterone)20Assessment of sperm motility:

the motility of each spermatozoon is graded :a =rapid progressive motilityb= slow or sluggish motilityc= non progressive motilityd= immotility

21Viabilityshould be determined if the % of immotile spermatozoa exceeds 50 %

Supravital stain: Eosin /NigrosinViable do not take up the stain

22Assessment of sperm morphology

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Normally the sperm count contains fewer than 20 % abnormal forms e.g. bitailed, short tailed , 2 heads ..etc.24Other cellular elements:

Leukocytes:

leukocytes predominantly neutrophils are present in most human ejaculate.Normal 40 WBCs/ HPF = ++If increased leukospermia or pyospermia may be associated with infection and poor sperm qualityWhen the number is increased, microbiological tests should be performed to investigate the presence of of infection in any of the accessory glands

25AgglutinationReported when motile sperm stick to each other in a definite pattern.

Head-headTail-tailHead-tail

Immunological cause of infertility

26Biochemical TestsFructose Test: It is secreted for sperm nutrition from seminal vesicleImpairment of seminal vesicular secretion will result in reduced fructose secretion in semen and the motility of the sperms will be reduced.

Fructose disappears in cases of : (a) absence of seminal vesicle;(b) obstruction of ejaculatory duct; and (c) inflammation of seminal vesicle. It is decreased in case of testosterone deficiency. So, fructose is used as fertility test. 27Acid phosphatase:

Secreted from the prostate. The test is used as: 1)A marker of prostatic functions; and 2)In forensic laboratories as a test for the presence of semen.

28Testing for Antibody coating of spermatozoaSperms can induce immune response not only between species but also within one species and within an individualsSperm antibodies may : Cause agglutination of spermCause reduction in motilityInhibit the ability of sperm to penetrate cervical mucusImpede binding of the sperm to the oocyteAntibodies may be IgA & IgG 29It must be remembered that sperm antibodies may be present in the cervical mucus itself , may inhibit the entry of sperm in the female genital system29Reference Ranges30Volume 2.0-6.0 mlpH 7.2-8.0Color: greyish whiteLiquefaction: 15-60 min.Count >20 million/mlTotal count > 40 million/ejaculateMotility: > 50 % motile( grades a+b) or > 25% with progressive motility ( grade a) within 60 minutes of ejaculatonMorphology > 30% normal formViability > 75% viableWBC< 1million/mlRBC none31Quoted from the WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction ,199931