Semen is a grey opalescent fluid which is formed at ejaculation. It is composed of suspension of spermatozoa in seminal plasma. Semen is made up of the secretion of all the accessory glands of the male genital tract:

(Testes 5%, Seminal vesicle 46-48%, Prostate 13-33%, Bulbourethral gland 2-5%). Semen analysis report include: Physical examination Microscopic examination Biochemical tests

1. PHYSICAL PROPERTIES OF SEMEN

normal value abnormal

VOLUME 2-5 ml/ejaculation Aspermia total absence of ejaculation (rare)

Oligospermia reduced volume of seminal fluid

Hypospermia seminal fluid volumeis less than 2 ml

Hyperspermia increased volume of semen above 10 ml (rare)

COLOUR greyish white -It is opalescent due to its high content of protein and the presence of more than 60 million sperms /ml

pale yellow discoloration

urine * easily detected by the consistency of the semen and the urineferous odour

pink blood (haematospermia) traces of fresh blood

bright red large amounts of blood

very bright yellow deep jaundice (bilirubin)

pH 7.3-8.1 * recorded on fresh semen by using pH paper with a range of 7-9

alteration of pH inflammatory conditions of prostate or seminal vesicles

VISCOSITY which allows semen to be poured drop by drop out of the container * measured the time taken by one drop to leave the standard pipette

SPECIFIC GRAVITY

1.028

2. MICROSCOPIAL EXAMINATION

normal value abnormal

SPERM COUNT = the number of sperms in an ejaculation

20 million/ml (about 60 millions/ejaculation) *obtained by multiplying the sperm concentration by the volume

Azoospermia no spermatocytes (male sterility)

Oligozoospermia <20 million/ml (<50million/ejaculation)

Polyzoospermia may reach 350 million/ejaculation

MOTILITY = percentage of sperms in the seminal fluid which are highly active

after 1 hour there must be >80% active sperms * performed soon after the production of the sample & is repeated after 1,2,3 and 6 hours after semen production

Grades of motility (WHO) A—Rapid forward progress motility B—Slow or sluggish progressive motility C—Nonprogressive motility D—Immotility the cutoff value for normal is - 50% grade A+B or - 25% grade A motility

Asthenospermia sperm motility less than the WHO cutoff levels

MORPHOLOGY normally, the sperm count contains <20% abnormal forms e.g. bitailed, short tailed, 2 heads, etc Non-sperm cells

RBCs - normally, there are no RBCs - if present, this indicates haematospermia

WBCs - normally there are very little number of WBCs - increase in cases of inflammation

epithelial cells always present in semen

pus cells - zero to few number -presence of large numbers of these cells indicates inflammation

spermatocytes (germinal cells)

- usually present in normal semen, but few in number

VIABILITY - when the motility is reported as less than 5% to 10%, viability testing is recommended because profoundly low motility may indicate dead sperm (necrospermia)

1. the most common viability assessment involves staining with Eosin Y followed by counter staining with Nigrosin the viable sperm with its intact cell membrane will not take up the dye and will remain unstained

2. Hypo-osmotic swelling test (HOST) - an alternative method to assess sperm viability - based on the principle that viable sperm have intact cell membranes

exposure of the sperm to hypo-osmotic fluid will cause water to flow into the viable cells seen as swelling of the cytoplasmic space & curling of the sperm tail

non-viable sperm with nonfunctional cell membranes will not exhibit this effect because they cannot maintain an osmotic gradient

3. BIOCHEMICAL TESTS properties/fx uses of tests/abnormal condition

FRUCTOSE - secreted from the seminal vesicle (150-650 mg%) - secreted for nutrition of sperm cells

- disappears in cases of: 1. absence of seminal vesicle 2. obstruction of ejaculatory duct 3. inflammation of seminal vesicle - decreased in case of testosterone deficiency fructose is used as fertility test using Seliwanoff’s test

ACID PHOSPHATASE

- secreted from the prostate

the test is used as: 1. a marker of prostatic functions 2. in forensic laboratories as a test for the presence of semen

ANTISPERM ANTIBODY TEST

approximately 10% of infertile men will present with antisperm antibodies (ASA) it has been suggested to be tested routinely in all men undergoing infertility work-ups

ACROSIN low acrosin activity has been associated with low sperm density, motility, and poor normal morphology

ZINC necessary for - chromatin stability and decondensation - head–tail detachment during fertilization

L-CARNITINE - secreted by the epididymis - concentrated in the seminal plasma at up to 10 times the serum levels - has a role in sperm maturation

- low L-carnitine levels are found in oligoasthenozoospermic men

ALPHA GLUCOSIDASE

- play a role in sperm maturation in the epididymis

- used to distinguish nonobstructive from obstructive azoospermia - used as a specific marker for epididymal function * a cutoff value of 12 mIU/mL distinguishes ductal obstruction from primary testicular failure

NORMAL SEMEN PARAMETERS

test normal value

LIQUEFACTION within 20 minutes MORPHOLOGY > 70% normal,mature spermatozoa

MOTILITY > 60% Ph > 7.0 (average 7.7%)

SPERM COUNT 60-150 million/ml VOLUME 1.5-5.0 ml

WBCs < 1million/ml

Precautions and steps of semen sample collection

There should be 2 to 7 days of sexual abstinence before collection The duration of abstinence should be constant, if possible

Two separate samples at least 7 days apart should be analyzed It is best to collect the specimen in a clean (not necessarily sterile), wide-mouthed jar It is important that the entire specimen be collected, because the initial fraction contains the greatest density

of sperm Ideally, collection should take place in the location where the analysis will be performed The degree of sperm motility should be determined as soon as possible after liquefaction, which usually

occurs 15 to 20 minutes after ejaculation Semen should not be exposed to marked changes in temperature, and if collected at home during cold

weather, the specimen should be kept warm during transport to the laboratory In order to allow liquefaction and mixing, semen is placed in a 370 C gently shaking incubator for 30 minutes The semen sample should be examined within 1 hour of production and receipt in the laboratory