SEMEN AND SEMINAL STAINS

SEMEN

- This is a viscid whitish fluid of the male reproductive tract consisting of spermatozoa suspended
in:
- The secretion of accessory glands

PARTS OF SEMEN

a. Seminal fluid
b. Epithelial cells
c. Crystals of choline and lecithin to 3.5 cc is the normal quantity of seminal fluid in single
ejaculation.

A healthy young man releases 400 t0 500 million spermatozoa in a single ejaculation.

CASES WHEREIN EJACULATION HAS NO SPERMATOZOA

1. Males suffering from aspermia

a. ASPERMIA
- A condition wherein males have no spermatozoa at all in their seminal fluid.
2. Males suffering from oligospermia
b. OLIGOSPERMIA
- Conditions whereby males have abnormally low sperm counts or with few spermatozoa.

WHERE SEMEN AND SEMINAL STAINS CAN BE FOUND

Semen can be found fresh in:

1. Vaginal contents of the victim

2. Rectal contents of the victim

Semen can be found as wet or dried condition/secretion on:

1. Hair
2. Skin around the genitals

Semen can be found as dry stain on:

1. Underclothing
2. Bed clothing

COLLECTION, PRESERVATION, PACKING AND TRANSPORT OF SPECIMEN:

1. Seizure of apparel must be done as soon as possible.

 2. In packing a clothing apparel worn by a victim, there should be no friction between the apparel
and the stain.
3. Specimen should not be rolled for transit.
4. Smaller objects like hair should be placed in a test tube and corked.
5. Specimen should be thoroughly dried before packing.
6. Fluid semen should be placed in a test tube. It may be preserved by a few drops of 10% solution
of formalin during hot weather.

SEMEN AND SEMINAL STAINS EXAMINATION

There are four examinations for seminal stains or seminal fluid in the form of stains, namely:

1. Physical Examination
2. Chemical Examination
a. Florence Test
b. Barberio’s Test
c. Acid-Phosphate Test
3. Microscopic Examination
4. Biological Examination

PHYSICAL EXAMINATION

SEMINAL STAIN

a. Semen when dry gives stiff, starchy feeling to the cloth and produces slight deeping of the color
with the disappearance of the color
b. Seminal stains exhibits fluorescence under the ultraviolet light.
c. Grayish white sometimes yellowish stain which is typical of seminal fluid.
d. They have an appearance or outline of a contour map.
e. May have a reddish tint in case of old man.

II. CHEMICAL EXAMINATION

A. FLORENCE TEST
This is only a preliminary test, presence of spermatozoa confirms the presence of seminal
stain.
REAGENT/CHEMICALS: Florence reagent (made of 1.65 g KI and grams I 2 in 30 cc water
Saline solution
PROCEDURE: Cut a portion of the stain and divide into small bits then soak in saline solution.
Transfer into a slide and evaporate the fluid.

Add a drop of Florence reagent and cover with cover slip. Examine under the microscope.
 VISIBLE OF POSITIVE RESULT: Crystals of choline per iodide that are dark brown, rhombic or
needle shaped that occur singly or in crossed even grouped in clusters.

B. BARBERIO’S TEST

This test is almost specific for human semen. Seminal stain as old as 6 years old are said to
respond to this test.

REAGENT/CHEMICALS: Saturated aqueous or alcoholic solution of picric acid.

PROCEDURE: Soak a piece of stained material in a 2.5% solution of trichloroacetic acid for
one hour on a test tube. Centrifuge the test tube. Get the clear liquid part and add to an
equal amount of saturated aqueous or alcoholic solution of picric acid on a glass slide.
Observe under the microscope.

VISIBLE OR POSITIVE RESULT: Crystals that are slender yellowish, rhomboid needles with
obtuse angle or appear as avoid crystals (these crystals are made of spermine picrate)

C. ACID-PHOSPHATASE TEST
This test is the best way to locate and at the same time characterize a seminal stain. This
test replaced the Florence test in reliability and was shown to be specific for human and
higher apes. The test is based fundamentally upon the extraordinarily high acid phosphatase
content of human male ejaculation.

REAGENTS/CHEMICALS: 23 g sodium chloride; 0.5cc of glacial acetic acid; 2 g of sodium

acetate trihydrate in 90 cc water; a suspension of 30 mg anthraquione-1-diazonium chloride
and 50 gms of calcium-1- alphanaphtyl phosphate in 1 cc of 1% aerosol.

PROCEDURE: Treat the stained area in a water bath at pH 5 containing alphanaphtyl

phosphate as substrate and antraquinone 1- diozorium chloride.. Add the above reagents.

VISIBLE OR POSITIVE RESULTS: Orange red pigment.

III. MICROSCOPIC EXAMINATION

The chief purpose of which is to determine the presence of spermatozoa. The

identification of spermatozoa is at present the only specific test for semen.
 VISIBLE RESULT: SPERMATOZOA

This is a small object with a pear-shaped head, behind which is a short neck and then a
tail about ten times as long as the head.

DETERMINATION OF SPERMATOZOA IN FRESH SEMEN

1. Transfer a drop of specimen to a glass slide.

2. Add a drop of water or saline solution and cover with cover slip.
3. Examine under the microscope
4. Observe for the presence of spermatozoa

DETERMINATION OF SPERMATOZOA IN SEMINAL STAINS:

1. A small piece of material is teased on a slide in a drop of water.

2. Allow the smear to dry and then stain with LOFFLER’S METHYLENE BLUE for a minute. Wash
with water, dry and examine under the microscope.

ELEMENTS WHICH MAY OBSTRUCT DETECTION OF SPERMATOZOA:

1. Nature of the fabric

2. Age of the stain
3. Conditions to which the stain was exposed before reaching the laboratory.
4. Handling of the specimen.

IV. BIOLOGICAL EXAMINATION

The test was proposed by Farnum in 1901.. He injected human semen to a rabbit from five to
eight times at intervals of 6 to 8 days. The serum obtained from the blood of the rabbit gave a
precipitate with quite recent and old emulsions of human semen.

Hektoen and Rustinant showed that this serum is both specie specific and semen specific, i.e. it
gives a positive reaction with human blood.

The spermato-precipitin is of value in the identification of seminal fluid in certain cases like for
example bestiality where it may be desirable to differentiate between the human seminal fluid from that
of the animal.
 LIMITATION:

The bacterial action which produces disintegration of spermatozoa in seminal stain is equally
effective in decomposing or digesting the protein constituents of semen that acts as the antigen-
producing antibodies. Such seminal stain with their protein constituents completely disintegrated
cannot possibly give a positive precipitin reaction.

OTHER STAINS OF MEDICO- LEGAL INTEREST

1. Obstetrical and gynecological stains

2. Excrements
a. Adults – yellowish brown and solid
b. Infant- greenish yellow
3. Faint stains
4. Rust stains
a. Rust – reddish-brown in color, insoluble in water and soluble in diluted acid resembles blood
stains
5. Synthetic Dye Stains
a. Resembles of old blood stain, but can be recognized by treating with strong acids and alkali.
6. Mineral Stains
a. These are due to red paints containing oxides of iron.
7. Stains of Vegetable Origin
a. Stain resembles blood. They may be produced by fruit juices like mulberry and mangosteen.

Almost all of the above mentioned stains can be different iated from blood stains through the
action of chemicals inasmuch as they result in a reaction that is not noticeable in blood.