BIOLOGICAL

EVIDENCE
(SEMEN)
 WHAT IS SEMEN?
• A COMPLEX MIXTURE PRODUCED BY THE MALE REPRODUCTIVE
ORGANS
• USUALLY WHITE TO YELLOWISH IN COLOR AND STICKY SUBSTANCE
• IT HAS A CHLORINE SMELL
• SLIGHTLY SWEET IN TASTE DUE TO HIGH CONTENT OF FRUCTOSE
• NORMAL QUANTITY IN A SINGLE EJACULATION IS 1.5 TO 5
MILLILITER
 COMPOSITION OF SEMEN
1. SPERM CELL OR SPERMATOZOA
A PEAR-SHAPED HEAD WITH A SHORT NECK AND A TAIL ABOUT TEN
TIMES AS LONG AS THE HEAD

2. SEMINAL FLUID
• THE FLUID PORTION OF SEMEN
• IT PROVIDES NUTRITIVE AND PROTECTIVE MEDIUM OF PROPER VOLUME
FOR CONVEYING THE SPERMATOZOA.
HTTPS://WWW.YOUTUBE.COM/WATCH?V=JQ5RVBJWFTQ
 SOURCES OF SEMEN
1. TESTIS
CONSTITUTES 2 – 5% OF THE SEMEN VOLUME. APPROXIMATELY
200 – 500 MILLION SPERM CELL PRODUCES IN THE TESTIS ARE RELEASED PER
EJACULATION
2. SEMINAL VESICLES
• APPROXIMATELY 65 - 75% OF THE SEMEN VOLUME IS DERIVED FROM
THE SEMINAL VESICLES.
• IT PRODUCES A YELLOWISH VISCOUS FLUID RICH IN FRUCTOSE,
VITAMIN C, PROSTAGLANDIN, PROTEIN KINASE, AND OTHER
SUBSTANCES WHICH NOURISH AND ACTIVATE THE SPERM
 SOURCES OF SEMEN
3. PROSTATE GLANDS
• CONTRIBUTES ABOUT 25-30% OF THE SEMEN VOLUME
• WHITISH (SOMETIMES CLEAR), ACIDIC FLUID CONTAINING ZIC,
PROTEOLYTIC ENZYMES, CITRIC ACID, ACID PHOSPHATASE AND
LIPIDS
4. BULBOURETHRAL GLANDS
• LESS THAN 1% OF THE SEMEN IS CONTRIBUTED BY THESE
STRUCTURES
 IMPORTANCE OF SEMEN ANALYSIS
• A DIAGNOSTIC ASSAY IN DETERMINING TREATMENT PLANS FOR INFERTILITY
• IMPORTANT IN ASSAULT CASES AS IT IS VERY USEFUL FOR DETECTION OF
SEMEN IN CLOTHS
• ISOLATION AND IDENTIFICATION OF SEMINAL STAIN FOUND ON VARIOUS
SUSPECTED AREA
• ANALYSIS OF VARIOUS PROTEIN PRESENT IN SEMEN OR SEMINAL STAIN
• SEMINAL STAINS ARE EXAMINED GENERALLY IN CASES OF SEXUAL
PERVERSIONS
• DETECTION OF SPERM CELL IN VAGINA AND OTHER BODY PARTS
• FECAL MATTER EXAMINATION
CASES WHEN SPERM CELL IS HARD TO DETECT

1. AZOOSPERMIA – TOTAL OR COMPLETE ABSENCE OF SPERM CELL

2. OLIGOSPERMIA – DEFICIENCY IN NUMBER OR ONLY FEW MOTILE
SPERM CELLS ARE PRESENT
3. NECROZOOSPERMIA – SPERM CELLS ARE PRESENT BUT ARE
COMPLETELY DEAD
 EXAMINATION OF SEMEN
1. PHYSICAL OR VISUAL EXAMINATION
• COLOR: YELLOWISH WHITE
• ODOR: DISTINCT MUSTY, UNPLEASANT OR CHLORINE LIKE AND KNOWN AS
SEMINAL ODOR
• TEXTURE: STRETCHY AND STARCHY
• APPEARANCE: USUALLY DIRTY HAVING VARIETY OF STAINS
2. MICROSCOPIC EXAMINATION
• MICROSCOPICALLY, DETECTION OF SPERM CELL IS ALSO CONSIDERED AS A
CONFIRMATORY EVIDENCE FOR THE PRESENCE OF SEMEN IN A SUSPECTED
STAIN.
 EXAMINATION OF SEMEN
3. PRESUMPTIVE TEST
A. ACID PHOSPHATASE TEST
• ONE OF THE BEST KNOWN AND MOST WIDELY EMPLOYED
TECHNIQUES FOR THE PURPOSE OF IDENTIFICATION OF SEMEN. THIS
TEST IS ADOPTED BECAUSE OF HIGH ACID PHOSPHATASE ACTIVITY OF
SEMEN.
• A POSITIVE RESULT SHOWS A PURPLE COLOR WHICH INDICATES THE
PRESENCE OF SEMEN
 EXAMINATION OF SEMEN
B. THIN LAYER CHROMATOGRAPHY
• THIS METHOD IS USED FOR THE DETECTION OF CHOLINE AND SPERMINE.
IN TLC, IT IS POSSIBLE TO DETECT CHOLINE AND SPERMINE
SIMULTANEOUSLY IN SEMINAL STAIN.
• IT IS POSSIBLE TO DETECT CHOLINE AND SPERMINE IN OLD SEMINAL
STAINS EVEN AFTER THE LAPSE OF 5 YEARS.

C. ULTRA VIOLET LIGHT EXAMINATION

• THE TEST WAS DEVELOPED TO DETECT BIOLOGICAL STAINS
• A VISUALIZED FLUORESCENCE UNDER UV LIGHT INDICATES PRESENCE OF
SEMINAL STAIN
 EXAMINATION OF SEMEN
3. CONFIRMATORY TEST
A. FLORENCE TEST
• THIS TEST WAS DISCOVERED BY DR. FLORENCE IN 1886
• A FLORENCE REAGENT IS APPLIED TO THE SLIDE CONTAINING THE STAIN AND A
POSITIVE RESULT SHOWS A DARK BROWN, RHOMBIC OR NEEDLE SHAPED
CRYSTALS
B. BARBERIO’S TEST
• INVENTED BY BARBERIO IN THE YEAR 1905
• WHEN THE QUESTION STAIN IS ALLOWED TO REACT WITH PICRIC ACID IT LEADS
TO THE FORMATION OF YELLOW NEEDLE SHAPED SPERMINE PICRATE CRYSTALS