Forensic examination of sample

Rohit Sah
 Specimen collection
• Specimen collected as part of forensic evidence include:

 Blood: For Drugs and DNA,

 Urine: Drugs, Pregnancy test
 Anal swab, Oral swab, Vaginal swabs: For Spermatozoa, DNA
 Swab from bite Marks: DNA
 Any stains: Blood or semen
 Nail clippings: skin cells, blood, fibre
 Pubic hair, Head hair: comparison study, DNA
 Skin : swab for Semen, saliva, blood; foreign material
 Clothing: Stains (Blood, saliva, semen), foreign material
Consent
• A written informed consent should be obtained before evidence collection.
• Informed consent must also be obtained for collection of samples for medical reasons e.g.
to draw blood samples for testing with STDs (Sexually Transmitted Diseases) as well as
for evidence collection e.g. DNA.

Precautions
• Proper technique used
• Collected as early as possible
• Labeling done accurately
• All wet specimens should be dried
• Specimen should be secure and tamper proof
• Chain of custody should be maintained
 Biological stains

• Biological stains are the body secretions or excretory products which leaves
its traces behind.
 Blood
 Semen
 Saliva
 Urine
 Feces
 Milk
 Vaginal secretions
BLOOD STAINS
 Collection of Blood Stains
• A clean piece of white filter paper or a piece of clean white cloth or gauge or cotton swab
may be used, allowing blood to soak into it, then drying it at room temperature. A control
filter paper, etc. should also be sent for examination.

• If the object is porous, a portion of unstained area should also be taken.

• If the object is non-porous and particularly if it is metallic, stains can be removed by

scraping and placed in small glass containers. They should not be placed in envelopes
where they will be reduced to powder.

• Stains on clothing may be scraped off or a fragment of the material cut.

• If blood is liquid, a sample can be pipetted and placed in a bottle and refrigerated.
 Preservation of Blood Stains

• The stained article is allowed to dry at room temperature.

• No extra heat should be used as this will cause deterioration of the stain.
• If the stained clothes are not dried, putrefaction sets in, and it becomes difficult or impossible
to know whether the blood is of human or animal origin.
• For packing bloodstained articles, polythene bags or air-tight containers must be avoided,
because the residual moisture may get trapped and give rise to bacterial growth leading to
destruction of the stains.
• Each garment should be wrapped separately and should not come in contact with other
clothing.
• While folding the garment, care should be taken to avoid two stains touching each other, as
they could belong to two different sources.
• A clean paper should be kept between each stain. All kinds of stains should be sent to the
State Forensic Science Laboratory for examination.
 Medico-legal Importance of Blood
1. Civil Importance
2. Criminal Importance

Civil Importance
• Disputed paternity and maternity issues
 Blood groups are hereditary factors and their modes of inheritance are helpful in disputed
paternity and maternity issues.

 It is possible to determine the groups of one parent if the groups of the child and the other
parents are known.

 Similarly, from the determination of blood groups of both parents it can be known to
possible blood groups for the child.

 When the same child is claimed by two women, or when two children are interchanged by
accident in maternity home, hospital etc., blood grouping tests are helpful.
• Divorce and nullity of marriage
 For example, presence of some forbidden diseases like Hemophilia,
Erythroblastosis foetalis etc.
 Note: Provision in Muluki Ain (Bihabari, no.4 and 5): nullity of marriage possible
between male and female if they are suffer from serious disease (Maharogi).

• Compensation cases
 Industrial/commercial workers exposed to different toxic agents. Examination of
blood may become important to determine the degree of harm suffered by the
workers due to a toxic agent.

• Civil negligence cases

 The questions arising in hospital/medical practices.
 E.g. Chemotherapeutic practices, blood transfusion etc.
Criminal Importance
• Blood is frequently accessible biological evidence in the various types of crime scene,
such as homicide, hit-and-run, rape, robbery etc.
• Depending upon the nature of case many questions may arises;
 Blood or other material?
 If, blood, Human or animal?
 If animal, Source of origin
 Arterial or venous, Victim or Accused, Ante mortem or postmortem, age of stain, injury
etc.
 Direction and heights of bleeding
 Cause of death.
 Sex of person
 Age of person
 Relationship between the offense, offender, victim and offending agents
 Any abnormalities of person
 Identification of person, accused, victim etc
Seminal fluid
 Introduction

• Semen contains sperm and secretions of seminal vesicles, prostate, Cowper's gland and
urethral glands.

• Identification of semen is important in sex-related crimes like rape, sodomy, incest and
sexual perversions.
 Collection of Material

• Fluid from the vagina is collected with a pipette or throat swab inserted with or without
the aid of a speculum, or vaginal washing i done,. which is concentrated by
centrifugation.

• Dried stains could be removed by scraping with a clean scalpel blade.

• Thin dried stain is collected with a wet throat swab.

• A portion of cloth containing the stain is cut out, dried and preserved.

• The pubic hair should be plucked and placed in a small container.

• Stains on smooth, impervious surface should be gently scraped off with the point of a
knife into a glass container.
 Preservation of Seminal fluid

• Seminal fluid should be handled promptly after collection to prevent degradation and
bacterial growth.

• It is recommended to transport the sample to the laboratory as soon as possible.

• Seminal fluid should be stored and transported at a controlled temperature. Generally,

refrigeration at 2-8°C (36-46°F) is suitable for short-term preservation.

• Avoid freezing the sample unless specific protocols require it, as freezing may
damage spermatozoa and affect subsequent analysis.
 Medicolegal Importance of Seminal fluid
• Sexual Assault Investigations: Seminal fluid analysis plays a crucial role in cases of
alleged sexual assault. The presence of semen in the victim's body or on clothing can provide
evidence of sexual contact or intercourse. DNA profiling from the semen sample can identify
the perpetrator, linking them to the crime scene or the victim's body.

• Paternity Testing: Seminal fluid analysis is essential for determining paternity in cases
where parentage is in question. By comparing the DNA profile of the child, the alleged
father, and the seminal fluid sample, it can be determined if the alleged father is biologically
related to the child.

• Infertility Investigations: Seminal fluid analysis is a fundamental diagnostic tool for

evaluating male infertility. Assessing semen parameters such as sperm count, motility,
morphology, and other factors can help determine the cause of infertility and guide
appropriate treatment.
• Sexual Offenses: Seminal fluid stains found at a crime scene or on the victim's body
can provide crucial evidence in sexual offense cases. The presence of semen can help
corroborate the victim's account and establish sexual contact or assault.

• False Allegations: In cases where false allegations of sexual assault are made, seminal
fluid analysis can play a critical role in establishing the absence of semen or disproving the
occurrence of sexual contact.

• Postmortem Examinations: Seminal fluid analysis may be performed in cases of

suspicious deaths or sexual assault-related homicides. It can provide valuable evidence in
determining the presence of sexual activity and help establish the circumstances
surrounding the death
Saliva
 Introduction

• Salivary stains may be found over clothes, body or private parts (breasts, face, neck),
unburnt portion of cigarette etc.

• Saliva stains are confirmed by presence of buccal squamous cells and/or a-amylase.

• It has significant role in sexual offence cases, identifying drug of abuse, blood groups
determination and sex determination.
 Collection of saliva

• Before collecting saliva, ensure that the individual has not eaten, drunk, smoked, or used
any oral hygiene products for at least 30 minutes prior to collection.
• Instruct the individual to rinse their mouth with water before collection to minimize
contamination.
• Provide clear instructions on how to collect the saliva, whether by drooling, spitting, or
using a collection device.
• Ensure that the collection container or device is sterile and suitable for preserving the
sample.
• Collect an adequate volume of saliva to ensure there is enough for analysis, typically
around 2-3 ml.
• Label the collection container or device with the individual's information, including their
name, date, and time of collection, to maintain proper identification.
 Preservation of Saliva
• Saliva samples should be handled promptly after collection to prevent degradation and
bacterial growth.

• It is recommended to transport the samples to the laboratory as soon as possible.

• Saliva samples should be stored and transported at a controlled temperature.

• Generally, refrigeration at 2-8°C (36-46°F) is suitable for short-term preservation.

• If long-term storage is required, samples can be frozen at -20°C (-4°F) or lower.

• However, note that repeated freezing and thawing should be avoided, as it may degrade
the sample.
 Medicolegal Importance of saliva
• DNA Analysis: Saliva contains epithelial cells from the lining of the mouth, which can be
a valuable source of DNA for identification purposes. DNA profiling from saliva samples can
help establish the identity of individuals involved in a crime, such as suspects, victims, or
unidentified individuals found at a crime scene.

• Sexual Assault Investigations: Saliva may be present in cases of sexual assault,

particularly in cases involving oral contact. The analysis of saliva samples can help identify
the presence of the perpetrator's DNA in cases where there was a forced or non-consensual
oral act.

• Bite Mark Analysis: Saliva can be present in bite mark cases, where the teeth of the
perpetrator leave marks on the victim's body. Saliva samples obtained from the bite marks
can help identify the individual responsible by analyzing the DNA present in the saliva.
• Drug Testing: Saliva can be used for drug testing purposes in cases such as driving
under the influence (DUI) investigations or workplace drug testing. Saliva samples can
detect the presence of drugs or their metabolites, providing evidence of recent drug use.

• Toxicology: Saliva can be analyzed for the presence of various substances, including
drugs, alcohol, toxins, and poisons. Saliva testing can be useful in cases of suspected
poisoning, substance abuse, or intoxication.

• Paternity Testing: Saliva samples can be used for paternity testing, where the DNA
profiles of the alleged father and child are compared to establish biological parentage.

• Disease Diagnosis: Saliva can be utilized for the diagnosis of certain diseases and
conditions. Saliva-based tests can detect the presence of specific markers or pathogens,
aiding in the diagnosis of oral diseases, infections, and other health conditions.
Hairs
 Collection of hairs
• Wear disposable gloves during the collection process to avoid contamination. Use clean,
sterile tools such as forceps or tweezers to handle the hair strands.
• Depending on the specific requirements, you may need to collect hairs from different
areas of the body
• Hair can be collected by plucking individual strands directly from the scalp or by cutting
close to the scalp using clean, sterilized scissors. Ensure that the entire length of the hair
from root to tip is obtained.
• About 20 hairs plucked or cut from the victim
• If root sheath attached, DNA analysis using PCR
• If there is no root sheath, microscopic analysis can detect characteristics as the victim’s/
suspect's hair
• Place the collected hair strands in a clean, sterile envelope or paper fold.
• Avoid using plastic bags, as they can trap moisture and cause degradation of the sample.
• Label the package with relevant information, including the individual's name, date of
collection, and the location of the hair sample.
 Preservation hair sample

• Place the collected hair sample in a clean, sterile envelope or paper fold. Avoid using
plastic bags, as they can trap moisture and cause degradation of the sample. Make sure
the packaging is properly sealed to prevent any loss or contamination of the hair.

• Hair samples should be protected from environmental contaminants, such as dust, dirt, or
other foreign substances. Handle the hair samples with clean, gloved hands to minimize
the risk of introducing extraneous DNA or other contaminants.

• Hair samples should be stored in a cool, dry environment to minimize the degradation of
the hair. Avoid exposing the samples to extreme temperatures, direct sunlight, or high
humidity, as these factors can negatively impact the stability and integrity of the sample.
 Medicolegal Importance of hairs

• Identification: Hair evidence can be used for individual identification purposes.

Comparison of hairs found at a crime scene with known hair samples from suspects or
victims can help establish their presence at the scene and potentially link them to the crime.

• DNA Analysis: Hairs with their roots intact contain DNA that can be extracted and
analyzed. DNA profiling from hair samples can provide valuable information for
identification, exclusion, or linkage in criminal investigations.

• Linking Individuals to a Crime: Hairs found at a crime scene, on a victim, or on an

object associated with a crime can potentially link a suspect to the scene. Comparison of
hair samples can establish associations between individuals, victims, and crime scenes.
• Sexual Assault Investigations: Hair evidence can be critical in sexual assault cases.
Hair transfer during sexual contact may link the victim and suspect. Comparing hair samples
from the victim's body or clothing with the suspect's hair can provide important evidence in
such cases.

• Drug Testing: Hair analysis can be used for drug testing purposes. It can provide a
historical record of drug use over an extended period. Testing the hair shaft can detect the
presence of drugs or their metabolites, providing evidence of past drug use.

• Cause of Death Determination: In cases of suspicious deaths, hair analysis can provide
valuable information about drug or toxin exposure, chronic substance abuse, or exposure to
environmental contaminants. It can contribute to determining the cause and manner of death.

• Species Identification: Hair analysis can aid in the identification of animal species, which
may be relevant in cases involving animal attacks or illegal wildlife trade.