Biological Body Fluids: Semen Analysis in Detail. Unit-V Hematology IVth Semester. DMLT

Biological body fluids

The various liquids within the human body that play essential roles in maintaining physiological functions are called Biological body fluids. The major biological body fluids include blood, plasma, serum, lymph, Cerebrospinal Fluid (CSF), Synovial Fluid, Pericardial Fluid, Peritoneal Fluid, Amniotic Fluid, and Sweat,

Semen Analysis in detail

Semen is the fluid released from the male reproductive system during ejaculation that consists of spermatozoa suspended in seminal plasma.

Significance of semen analysis

• 1. Semen analysis provides information about male fertility. It identifies problems related to sperm production and function.

• 2. Semen analysis is a must to examine vaginal secretions or clothing stains for the presence of semen in rape.

• 3. Semen analysis helps to check the effectiveness of vasectomy.

• 4. Cervical mucus from a female partner is examined for viable spermatozoa.

Collection of specimen

• 1. Instruct patient for 5 to 7 days of abstinence.

• 2. Collect the specimen in a clean dry wide-mouth container by masturbation. Collection of specimens by coitus interruption is not a satisfactory procedure.

• 3. Examine the specimen within 2 hours of ejaculation.

Components of Semen Analysis

Physical and chemical examination of semen

• 1. Volume: Normal volume ranges typically fall between 1.5 and 5 ml.

• 2. Viscosity: Fresh specimen is highly viscid, opaque, white, or grey-white. Shelf liquification occurs within 15 to 30 minutes. Absence of liquification affects the spermatozoa movement in the vaginal canal. Prostatic enzyme initiates liquefaction.

• 3. pH (Acidity/Alkalinity): Semen pH measures the acidity or alkalinity of the semen. Normal pH levels are typically between 7.2 and 8.5 with an average of 7.7. Abnormality due to the pH of semen is very rare.

• 4. Fructose Level: Fructose is a sugar that provides energy for sperm. A low fructose level may indicate a problem with the seminal vesicles.

Procedure: Add 5.0 ml resorcinol reagent in a test tube. Add 5.0 ml semen specimen. Mix resorcinol and semen specimens properly. Place the tube in a boiling water bath for 5 minutes.

Observation: No color change indicates the absence of fructose. The appearance of red precipitate within 30 seconds indicates the presence of fructose in the semen specimen.

Microscopic Examination of semen

1. Sperm Concentration (Sperm Count): Sperm concentration measures the number of sperm cells per milliliter of semen. A normal sperm concentration is 60 to 150 million sperm/ml. It may go up to 300 million sperm/ml.

Process: Allow liquefaction of semen. Mix the semen sample with diluting fluid in a ratio of 1:20. Use a WBC pipette. For mixing, draw semen in a WBC pipette up to mark 0.5. Draw diluting fluid in the WBC pipette up to mark 11. Use the Neubauer counting chamber to count several sperms.

Calculation: Count spermatozoa in 2 sq mm and multiply it with 100000. This gives the number of spermatozoa per ml of semen.

2. Motility of spermatozoa: Sperm motility refers to the percentage of sperm that are actively moving. There are two types of motility. Progressive Motility: Sperm that moves forward in a straight line. Non-Progressive Motility: Sperm that move, but not in a straight line.

Place a small drop of semen on a clean dry glass slide. Cover it with the coverslip. Examine it under high power objective and low power objective. Observe at least 200 spermatozoa. Check thre proportion of motile and non motile spermatozoa.

3. Sperm Morphology (Shape and Size): Morphology evaluates the shape and structure of sperm. The percentage of sperm with normal morphology is usually above 4%.

Process: Prepare a semen smear on a dry clean glass slide using one drop of semen. Stain the smear using a suitable stain solution. The best staining method is the hematoxylin method or the Papanicolaou method. Other stains like basic fuchsin, Giemsa, and crystal violet are also used.

Staining results:

• Nucleus of head: Dark blue

• The cytoplasm of the head: Pale blue

• Middle piece&Tail: Pink-red

Observation: Normal spermatozoa are 50 to 70 micrometers in length. It has a large head, a small neck, and a long tail. 90% of spermatozoa body consists of tail.

The normal value of sperm motility is 80 to 85%.

4. Total Sperm Count: This parameter is calculated by multiplying the sperm concentration by the semen volume. The total sperm count provides an overall assessment of the number of sperm in the entire ejaculate.

5. White Blood Cells (WBCs): The presence of white blood cells in semen may indicate an infection or inflammation. Elevated levels may require further investigation.

STRUCTURE OF SPERMATOZOA (Sperm):

sperms are microscopic and highly motile male gametes. Sperms can survive inside the female reproductive system from 24 hours to 48 hours. Human sperm consists of four parts: Head, Neck, Body, and Tail.

1. Head: It is the anterior-most, enlarged, and flat part of sperm. The Head has two components. These are the large nucleus and cap-like structure acrosome. The acrosome has sperm lysine. Sperm lysine is used to contact the ovum and to penetrate the ovum during fertilization.

2. Neck: It is a narrow part present between the head and body of sperm. The neck has two centrioles in its cytoplasm. The proximal centriole is free and present near the nucleus.

3. Body: It is also called the middle piece of sperm. It is present in between the neck and tail. Axial filament passes through this middle piece and it is coiled by mitochondria called mitochondrial spiral. The mitochondrial spiral provides energy for sperm movement and vibration.

4. Tail: It is the longest part of the sperm. The axial filament runs through the tail and is surrounded by a thin layer of cytoplasm. The tail has two parts main piece and the end piece. The main piece becomes narrow gradually towards the posterior end This end piece of the tail is considered as the naked end of the axial filament. The tail vibrates very rapidly that helps sperm to swim in a fluid medium.

Interpretation of Results Abnormalities

Microscopic examination may observe the following abnormalities in semen

• 1. Aspermia: Complete absence of spermatozoa in multiple slides of semen.

• 2. Oligospermia: Presence of only a few spermatozoa in semen during semen analysis.

• 3. Necrospermia: Presence of plenty of spermatozoa in semen but all are nonmotile.

• 4. Haemospermia: Presence of a large number of red blood cells and pus cells along with spermatozoa in semen.

Abnormalities in sperm morphology

• 1. Spermatozoa: Abnormally shaped head of spermatozoa, Abnormal size of spermatozoa head, double-headed spermatozoa, thin-headed spermatozoa, coiled tail large-headed. spermatozoa. Absence of neck, swollen neck, absence of tail, short tail, double tail, or terminal droplet tail

• 2. Others: Presence of red blood cells, pus cells (leukocytes), epithelial cells, bacteria, or fungus.

Abnormal spermatozoa should not be more than 20% of all spermatozoa present in the semen.

Dr Pramila Singh