IDENTIFICATION OF BLOOD AND BLOOD STAINS

Legal Importance of the Study of Blood:

1. For disputed parentage (maternity and paternity):
a. Disputed paternity may arise:
(1) When the wife committed adultery and the husband denied to be the father of
the child.
{2) When a child was born out of lawful wedlock and the mother claimed
someone to be the father but he vehemently denied it.
(3] In a claim for support or right of succession of the alleged illegitimate child,
b. Disputed maternity may arise:
(1) In case of allegation of jnterchange of children in a hospital or nursery home,
either accidentally or deliberately.
(2) In cases of wayward or stray children being claimed by two or more women.
(3) For ownership of dead fetus or newly born child found in street trash.

2. Circumstantial or corroborative evidence against or in favor of the perpetrator of a crime:

Example: "A" was found dead with a deep stab wound on the chest. "B" was found with a
kitchen knife in his hand stained with blood. Examination of the weapon showed that the stain
was blood of human origin and belonging to the same group as that of the deceased "A". With
such result of the examination, the investigating authorities have a very strong presumption
that "B" was the one who committed the crime.

3. Determination of the cause of death:

The amount of blood or blood stains found in the scene of the crime or found inside the body
of the deceased outside the blood vessels may imply that the cause of death of the person is
hemorrhage.

4. Determination of the direction of escape of the victim or the assailant:

The shape of the blood or blood stains will give the investigator an idea on the direction of the
source of blood. Usually, in small drops, the tapering end of the blood spot is towards the
direction of the moving source of blood.

5. Determination of the approximate time the crime was committed:

Although there are variations as to the color and soluble changes as to regards the age of the
stain, we can only say that when there is too much change, it is not very recent.

6. Determination of the place of commission of the crime.

7. Determination of the presence of certain diseases.

Problems to be Answered in the Examination of Blood:

1. Determine whether the stain is due to blood.
2. If due to blood, determine whether it is of human origin or not.
3. If it is of human origin, to what group does it belong?
4. 4. Does it belong to the person in question?
5. The manner, degree and condition of the article which have been stained.
6. Age of the stain.

PRELIMINARY OR GROSS EXAMINATION OF THE STAIN:

1. Determine the material, make, color of the article stained.
2. Note which surface has been stained and the color of stain. Recent blood stains are dark-
red.
3. Study the direction of the origin of the blood stain.
The spot of blood is usually tapering towards the direction of the source. A fall will give a
splash appearance.
4. For small and discolored stains, the use of a lens or ultra-violet light may be useful.
5. Determine the amount by the degree of soaking, size and intensity of color.

PHYSICAL EXAMINATIONS:
1. Solubility test:
Recent blood shed is soluble in saline solution and imparts a bright red color. Stains
which have been exposed to air become dry; hemoglobin is transformed to meth-
hemoglobin or hematin. If the stain has been kept in damp places for a long time;
hemoglobin is transformed to hematin.

2. Heat test:
Solution of the blood stain when heated will impart a muddy precipitate.

3. Luminescence test:
Stains on dark fabric mixed with mud, paint, etc. emit bluishwhite luminescence in a
dark room when sprayed with one of the two solutions:
a. 3-amino-phthalic-acid-hydrazide-HCL 1 gram Sodium peroxide 5 grams Distilled water
1,000 cc.
b. 3-amino-phthalic-acid-hydrazide-HCL 1 gram Sodium carbonate 50 grams Hydrogen
peroxide (10 Vol.) 50 grams Distilled water 1,000 cc.

The substance responsible for the reaction is hematin. Older stains therefore react
better than new ones. Although the solutions are said not to interfere with further tests,
unsprayed specimen of the material must be kept for the serologic test. (Lyon's Medical
Jurisprudence for India by S.P.S. Greval, 1953, p. 303).

CHEMICAL EXAMINATIONS:

1. Saline extract of the blood stain plus ammonia will give a brownish tinge due to the
formation of alkaline hematin.

2. Benzidine test: A piece of white filter paper is pressed firmly on the suspected stain.
Benzidine reagent is dropped on the paper, then followed by drops of active hydrogen
peroxide. A positive result will show blue color. A positive result is not conclusive,
because an oxidizing agent will give a positive blue color reaction. Benzidine test has the
sensitivity up to 1:300,000 dilution.

Benzidine reagent: Benzidine sulphate is dissolved in glacial acetic acid to form 10%
solution.

3. Guaiacum test (Van Deen's Dyas' or Schombein's Test):

To a white filter paper pressed and rubbed on the surface of the stain, the solution of
the alcoholic tincture of guaiacum is added and then hydrogen peroxide or ozonic ether
is applied by drops. If blood is present, a blue color is imparted by the mixture. It is not
conclusive like the benzidine test because potato skin, iron rust, cheese, blue and indigo
may give a positive reaction to the test. The guaiacum test is positive up to 1:5,000
dilution.

4. Phenolphthalein test (Kastle-Meyer Test):

A drop of the Kastle-Meyer's reagent is dropped on a white filter paper with the stain
and left for at least ten seconds. A positive result will show a pink color after the
addition of hydrogen peroxide. This test is not conclusive but sensitive up to
1:80,000,000 dilution. This test proves only the presence of peroxidase. Kastle-Meyer's
reagent:

5. Leucomalachite Green test:

This test which was recommended by Adler in 1904 is quite useful, but it is not so
sensitive as the benzidine test. It depends upon the fact that leucomalachite green is
oxidized to malachite green with a bluish-green or peacock-blue color by hydrogen
peroxide solution. The reaction occurs also with a solution of the blood pigment
previously boiled. On the other hand, the reaction is negative when iron is removed
from hemoglobin forming hematoporphyrin.

MICROSCOPIC EXAMINATIONS:
Saline extract of the stain is examined under the microscope. Note the presence of red
blood cells, leucocytes, epithelial cells and microorganisms. The presence of red blood
cells will conclusively show that the stain is blood.
By microscopic examination, we can differentiate the origin or the part of the body it
came from. Menstrual blood will show abundance of vaginal epithelial cells and
Doederlein's bacilli.

MICRO-CHEMICAL TESTS:
1. Hemochromogen crystal or Tokayama test:
2. Teichmann's blood crystals or Hemin crystals test:
3. Acetone-haemin of Wagenhaar test:
SPECTROSCOPIC EXAMINATIONS:
This examination depends on the principle that blood pigments have the power to
absorb light of certain wave length and produce certain characteristic absorption bands
on the spectrum. By means of the spectroscope we can determine the presence of the
following substances:
1. From fresh blood stains:
a. Oxyhemoglobin
b. Hemoglobin
c. Reduced hematin or hemochrogen
2. From older stains:
a. Methemoglobin
b. Alkaline hematin
c. Hematoporphyrin
d. Reduced hematin
3. Other blood preparations:
a. Acid hematin
b. Alkaline hematin
c. Carboxyhemogiobin
d. Hematin

BIOLOGIC EXAMINATIONS:

1. Precipitin test:
This test is to determine whether the blood is of human origin or not. Principle of the
test: By injecting an animal, usually, a rabbit, with defibrinated blood of unrelated
animal, an anti-serum is produced in the blood of the animal injected. The serum of this
animal injected is capable specifically of precipitating the serum of the unrelated animal
whose blood serum has been injected. However, closely related animals may also give
the same response.
Value of the precipitin test: If positive result is obtained, we can tell in a more or less
conclusive way that the blood stain is of human origin; although anthropoid ape may
give the same result. The same test and technique may be made to determine whether
muscles, secretions, bones and other body fluids are of human origin or not. Certain
materials like alcohol, formaldehyde, corrosive sublimate, lysol, creoline, carbolic acid,
acids and alkalies destroy the property of blood to react with precipitin.

2. Blood grouping:
Principle of the test: All human beings have their blood belonging to any of the four
principal blood groups. A normal suspension of human red blood cells when mixed with
its own serum or serum of a similar group will make the red blood cells suspension
remain even. But if suspended in the serum of another group, the red blood cells clump
with one another and this is called agglutination. The red blood cells contain
agglutinogens and the serum contains agglutinins.

Value of the test: It may solve disputed parentage (paternity or maternity). A positive
result is not conclusive that the one in question is the offspring, but a negative result is
conclusive that he is not the child of the alleged parents.

Age of the Blood Stains:

When blood is exposed to the atmosphere or some other influences, its hemoglobin is
converted to meth-hemoglobin or hematin. The color is changed from red to reddish-
brown. The presence of acid accelerates the formation of hematin. These changes take
place in warm weather within 24 hours. Blood of one week old and that of six weeks
may not present a difference in physical and chemical properties.

Differential Characteristics of Blood from Different Sources:

1. Arterial Blood:
a. Bright scarlet in color.
b. Leaves the blood vessel with pressure.
c. High oxygen contents.
 2. Venous Blood:
a. Dark red in color.
b. Does not spill far from the wound.
c. Low oxygen content.

3. Menstrual Blood:
a. Does not clot.
b. Acidic in reaction owing to mixture with vaginal mucous.
c. On microscopic examination, there are vaginal epithelial cells.
d. Contains large number of Deoderlein's bacillus.

4. Man's or Woman's Blood:

There is no method differentiating a man's blood from a woman's blood. Probably, the
presence of sex hormone in female blood may be a point of differentiation.

5. Child's Blood:
a. At birth, it is thin and soft compared with that of adult.
b. Red blood cells are nucleated and exhibit greater fragility.
c. Red blood cells count more than in adult.

Normal Anatomy
Anatomy is the identification and description of the structures of living things. It is
a branch of biology and medicine.

The word “anatomy” comes from the Greek words “ana,” meaning “up,” and “tome,”
meaning “a cutting.” Studies of anatomy have traditionally depended on cutting up, or
dissection, but now, with imaging technology, it is increasingly possible to see how a
body is made up without dissection.
There are two ways of looking at anatomy: Gross, or macroscopic, anatomy and
microscopic anatomy.

1. Gross anatomy

In medicine, gross anatomy, macro anatomy, or topographical anatomy refers to the

study of the biological structures that are visible to the naked eye.

The study of gross anatomy may involve dissection or noninvasive methods. The aim
is to collect data about the larger structures of organs and organ systems.

In dissection, the scientist cuts open the human or animal cadaver is cut open and
examines its organs.

2. Endoscopy

Inserting a tube with a camera at the end, might be used to study structures
within living animals. Endoscopy is performed either through the mouth or
through the rectum, so the gastrointestinal tract is often the primary organ of
interest.

There are also less invasive methods. For example, to study the blood vessels of living
animals or humans, the scientist may insert an opaque dye into the animal that will
highlight the circulatory system when imaging technology, such as angiography.

Techniques such as magnetic resonance imaging (MRI), computed tomography (CT),

or x-ray also reveal information about the inside of a living body.

Medical and dental students perform dissection as part of their practical work in gross
human anatomy during their studies. They may dissect a human corpse.

Students of gross anatomy will need to learn about the major body systems.
Human body systems

There are 11 organ systems in the human body:

 The integumentary system, meaning skin, hair, nails, and so on

 Skeletal system
 Muscular system
 Lymphatic system
 Respiratory system
 Digestive system
 Nervous system
 Endocrine system, which regulates hormone production
 Cardiovascular system
 Urinary system
 Reproductive systems

Although these systems have different names, they all work interdependently, meaning
they work together and depend on each other.

Microscopic anatomy

Microscopic anatomy, also known as histology, is the study of cells and tissues of
animals, humans and plants that are too small to been seen with the naked eye.

By looking at tissue under the microscope, we can learn about the architecture of the
cells, how they are put together, and how they relate to each other.

For example, if a person has cancer, examining the tissue under the microscope will
reveal how the cancer cells are acting and how they affect normal human tissue.

This commonly involves studying tissues and cells using histological techniques such
as sectioning and staining, and then looking at them under an electron or light
microscope.
Sectioning involves cutting tissue into very thin slices so they can be examined.
Histological stains are added to biological structures, such as tissues, to add colors or
to enhance colors so they can be more easily distinguished when they are examined,
especially if different structures are next to each other.

Histology is vital for the understanding and advancement of medicine, veterinary

medicine, biology, and other aspects of life science.

Histology is used for:

Forensic investigations: The microscopic study of biological tissues can help explain
why, for example, somebody unexpectedly died.

Autopsies: As in forensic investigations, biological tissues from deceased people and

animals can be analyzed, so that investigators may better understand the causes of
death.

Vital Signs
There are four main vital signs: body temperature, blood pressure, pulse and breathing rate.
Normal ranges for these signs vary by age, sex, weight and other factors.
What are vital signs?

Vital signs are used to measure the body's basic functions. These measurements are taken to
help assess the general physical health of a person, give clues to possible diseases and show
progress toward recovery. The normal ranges for a person's vital signs vary with age, weight,
gender and overall health.

There are four main vital signs: body temperature, blood pressure, pulse (heart rate), and
breathing rate.

Body temperature: 

A drop in body temperature below 95º Fahrenheit is defined as hypothermia. Keep in

mind that temperature can vary due to factors other than illness or infection. Stress,
dehydration, exercise, being in a hot or cold environment, drinking a hot or cold
beverage and thyroid disorders can influence body temperature. Because older adults
do not control body temperature as well as younger adults, older adults may be ill
without ever displaying signs of a fever.
Blood pressure: Blood pressure is the measurement of the pressure or force of
blood against the walls of your arteries. Blood pressure is written as two
numbers, such as 120/80 millimeters of mercury (mm Hg). The first number is
called the systolic pressure and measures the pressure in the arteries when the
heart beats and pushes blood out to the body. The second number is called the
diastolic pressure and measures the pressure in the arteries when the heart rests
between beats.

Pulse: Your pulse is the number of times your heart beats per minute. Pulse
rates vary from person to person. Your pulse is lower when you are at rest and
increases when you exercise (because more oxygen-rich blood is needed by the
body when you exercise). A normal pulse rate for a healthy adult at rest ranges
from 60 to 80 beats per minute. Women tend to have faster pulse rates than
men. Your pulse can be measured by firmly but gently pressing the first and
second fingertips against certain points on the body — most commonly at the
wrist or neck (but can also be measured at the bend of the arms, in the groin,
behind the knees, inside the ankles, on the top of the feet or at the temple area of
the face) — then counting the number of heart beats over a period of 60
seconds. A faster than average pulse can indicate such health problems as
infection, dehydration, stress, anxiety, a thyroid disorder, shock, anemia or
certain heart conditions. 

Respiratory rate: A person's respiratory rate is the number of breaths you take
per minute. The normal respiration rate for an adult at rest is 12 to 20 breaths per
minute. A respiration rate under 12 or over 25 breaths per minute while resting is
considered abnormal. Among the conditions that can change a normal
respiratory rate are asthma, anxiety, pneumonia, congestive heart failure, lung
disease, use of narcotics or drug overdose.