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Forensics &Bioterrorism
Arguilles | Benig | Guiriba | Navarro | Tampil
FORENSICS
OUTLINE
I. INTRODUCTION
A. BRIEF HISTORY
B. AREAS IN FORENSIC SCIENCE
C. BASICS OF CRIME SCENE INVESTIGATION
II. TECHNIQUES AND METHODS USED
III. EXAMPLES
IV. RECENT PROGRESS AND ISSUES
WHAT IS FORENSIC SCIENCE?
Science is a systematically organized body of knowledgeForensic is derived from the latin word, forum which means “public”
Forensic science - “The science of associating people, places, and
things involved in criminal activities”
Houck and Siegel, 2010
AREAS OF FORENSIC SCIENCE
CriminalisticsCollects and analyzes physical evidences
Forensic PathologyDetermination of cause and manner of death
Forensic AnthropologyIdentification of human remains not possible through soft tissue features
Forensic OdontologyAlso called forensic dentistry
Forensic Engineeringinvestigate and tests materials, products or structures that do not function the way they used to
Forensic ToxicologyDetermination of drugs present in body fluids and tissues
Behavioral Sciences
18TH & 19TH
CENTURY
SCIENCE OF PERSONAL
IDENTIFICATION
- STUDY OF FINGERPRINTS
- FORENSIC BLOOD TYPING
- FORENSIC DNA TECHNOLOGY
EARLY 20TH
CENTURY
COMPARISON
MICROSCOPE
5TH CENTURY
FORENSIC MEDICINE
TOXICOLOGY AS ONE OF THE
NEW EMERGING FORENSIC
SCIENCES
TOXICOLOGY AS ONE OF
THE NEW EMERGING
SCIENCES
BRIEF HISTORY
Houck and Siegel, 2010
FORENSIC SCIENCE begins at the CRIME SCENE
CRIME SCENE INVESTIGATION TEAM
(Jackson & Jackson, 2017)
Securing the scene
Safety and preservation of evidences
Separating the WitnessesGathering
information and Comparison
Scanning the Scene
Determining where photos
should be taken
Seeing the Scene
Photography for present &
future use
Sketching the Scene
Searching for EvidencesGathering information and
Comparison
Securing & Collecting Evidence
Preserving Integrityof Evidences(Placement ofEvidence at X)
12 3 4
5 6 7
CRIME SCENE INVESTIGATION
(Bertino & Bertino, 2012; Jackson & Jackson, 2017)
PHYSICAL EVIDENCESEvidence address to the senses of the court that are capable of being exhibited, examined, or viewed by the court (PNP, 2011)
HAIR BLOOD FINGERPRINT
BODY FLUIDS FIBER GLASS(Bertino & Bertino, 2012; Jackson & Jackson, 2017)
HOW DNA TESTING WORKS- A person’s DNA is unique in every person.
-Only identical twins share the same DNA.
-Scientists identify alleles (small portions of the DNA) & give them a unique number representing a person’s DNA profile.
-At each DNA location a person has 2 alleles (GC-AT)
-Paternity Testing: half of the DNA of a child comes from the mother & half comes from the father
-Potential Sibling: biologically related individuals have more DNA in common than those who don’t (Bertino & Bertino, 2012; Jackson & Jackson, 2017)
ANALYSIS OF EVIDENCES
DNA PROFILING & FORENSIC DNA PHENOTYPING
DNA PROFILING
Comparison of DNA in nucleated cells of person with biological matter found at the
crime scene or with the DNA of another person for the purpose of identification or
exclusion
The Modern DNA fingerprinting procedure or DNA typing
Allows a biological sample at a scene of crime or accident to be linked very strongly to the individual from whom it originated
(Jackson & Jackson, 2017; Jeffreys et al, 1985)
DNA PROFILING PROCEDURE
Extract DNA from
the Sample
DNA Quantitation
PCR Amplification Electrophoresis
Pattern Interpretation &
Matching
-Centrifugation-Agitation-Removal of proteins, lipids & RNA- DNA Purification
- Human DNA Isolation- Quantifiler DNA Quantification Kit-ABI PRISM 7500 Sequence Detection System
Amplification of genetic loci using PCR for forensic analyses
(Bertino & Bertino, 2012; Jackson & Jackson, 2017)
DNA PROFILING TECHNIQUES
Only about 1 ng (10¯⁹ g) of DNA is required for an optimum DNA profiling result. OR LESS.
RFLP(Restriction Fragment Length
Polymorphism)
PCR(Polymerase Chain Reaction)
SNP(Single Nucleotide
Polymorphism)
STR & VNTR(Short Tandem Repeats & Variable
Number Tandem Repeats)Y-chromosomal AnalysisMitochondrial DNA Analysis
(Sharma & Vidya, 2016)
-The in vitro replication of DNA fragments-Used when specimens are very little or the DNA is degraded
PRINCIPLE: Amplification of the biological process of DNA replication but is confined to desired specific DNA sequences.
METHOD:1. Denaturation – 94-96 °C , 5 mins & 30 sec2. Annealing – 62 °C, 30 sec3. Polymerization – 72 °C, 30 sec
Polymerase Chain Reaction (PCR)
(Jackson & Jackson, 2017; Sharma & Vidya, 2016)
Polymerase Chain Reaction (PCR)
DNA Profile of 4 persons using 10 primers (reverse and forward)
Restriction Fragment Length Polymorphism (RFLP)- Variation in the length of a DNA fragment produced by a specific
restriction enzyme acting on DNA from different individuals. Variation usually results from a genetic mutation.
- PRINCIPLE: Restriction enzymes recognize very specific restriction sites in a DNA sequence. Individuals rarely have the same array of restriction sites and distances in every DNA sequence.
Restriction Enzymes
(Losos et al, 2007; Sharma & Vidya, 2016)
Restriction Fragment Length Polymorphism (RFLP) - METHOD:
Restriction Digestion
Restriction enzyme cuts specific restriction
site in the DNA
Gel Electrophoresis-DNA fragments are separated by
length & size-Movement towards (+) Electrode-Smaller fragments move faster
Southern BlottingssDNA fragments migrates
from gel to nylon membrane using electric current
ProbingSLP or MLP
Membrane is incubated with radioactive
fluorescent strand of DNA
HybridizationBinding of DNA fragment with
probeNOTE: Probe will only bind to complementary parts, others
are washed off
Developing Probe Pattern/Autoradiogram
Nylon membrane placed againstX-ray Fillm
Image formation by particle emission of radioactive DNA
1 2 3
4 5 6
(Losos et al, 2007; Sharma & Vidya, 2016)
Restriction Fragment Length Polymorphism (RFLP) -PROBING: Locus serves as the target area of DNA
-DNA SLP or MLP band patterns from stains of blood or sperm can be compared with patterns from persons suspected of leaving the stains.
-Limitation of MLP in statistical interpretation of the multi-banded pattern formed. With this, in 1990, MLP was replaced by SLP.
Multiple Locus Probe (MLP) a probe that hybridizes to a
number of different sites in the genome of an organism
Single Locus Probe (SLP) a probe that is able to hybridize
with DNA from a specific restriction fragment on the
Southern blot.
(Losos et al, 2007; Sharma & Vidya, 2016)
Restriction Fragment Length Polymorphism
(RFLP) - METHOD:
(Giuseppe et al, 2002)
Single-Nucleotide Polymorphism (SNP)-Represents the alterations in the DNA sequence at a single
nucleotide position, either due to base changes, insertion or deletions of one or few bases.
-PRINCIPLE: Alterations create different alleles of that gene.Mutation in SNP is very rare, the sequence tend to be passed unchanged across generations.
-Difference in the sequence itself rather the length of DNASNP occurs every 100-300 bases along entire length of the human genome. - DNA template can be as large as a pair of specific primers ~50bp compared to ~300bp needed in STR. (Sharma & Vidya, 2016)
Short Tandem Repeats (STR) & Variable Number of Tandem Repeats (VNTR)-Are DNA sequences whose length varies from 1,000-20,000 bases and contains
repeating sequences for every 2-9 base pairs (STR) or 10-100 bp (VNTR).
-Number of repeat units vary between individuals.
-Cannot take place without the presence of specific oligonucleotides that serves as primers for the DNA polymerase and deoxynucleotide triphosphates, thebuilding blocks of DNA.
-STR is a.k.a. microsatellite
(Sharma & Vidya, 2016)
STR are dispersed more evenly than VNTR in the genome
Short Tandem Repeats (STR) & Variable Number of Tandem Repeats (VNTR)
(Jackson & Jackson, 2017;Sharma & Vidya, 2016)
-Analyses based on STR Profiles
1. Y-Chromosomal Analysis- Y-STR typing uses Y-STR markers give specific
information about the presence of male DNA.- Useful for rape cases and male lineage
2. Mitochondrial DNA Analysis-Contains genetic material found only from the
mother-Enables to type ancient samples with only minute
amounts of DNA needed-mtDNA is present in large amounts in the cell and
are highly stable. (ex. Romanov Case)
COMPARISON OF DNA PROFILING TECHNIQUES
(Sharma & Vidya, 2016)
Other Uses of DNA: FORENSIC DNA PHENOTYPING (FDP)
-Makes use of DNA genotype for the prediction of appearance traits (phenotype) of missing individuals or suspects.
-Results of FDP can serve as “biological witness” wherein Externally Visible Characteristics of a missing person or possible perpetrator can be inferred, however remains an issue.
(Kayser, 2015)
CASES OF DNA PROFILING
CASES INVOLVING FORENSIC DNA ANALYSIS
Pitchfork Case (1987) Murder, Narborough, Leicestershire, EnglandFirst case that a man (Colin Pitchfork) was convicted of murder and rape through the aid of DNA profiling of blood & saliva samples and comparing it with the DNA of semen evidence.
Routier Case (1996)Murder, Rowlett, TexasDNA profiled from blood evidences using STR and Y-STR techniques proved that it was the mother (Darlie Routier) who killed her two sons (Devon and Damon) and not an intruder which she initially stated.
(Elvidge, 2016; HubPages Inc., 2017)
CASES INVOLVING FORENSIC DNA ANALYSIS-In the Philippines,
Vallejo (2002) & Yatar (2004) CasesRape-Homicidewere solved after finding DNAs of the perpetrators (Gerrico Vallejo, Joel Yatar) the vaginal swabs of the victims. Note: The 2 are separate cases.
De Villa Case (2001) Rapewas reopened in 2003 for DNA-based paternity testing finding out that the DNA of the child of the said raped victim (niece of the suspect) did not match with the perpetrator
(De Ungria, n.d.)
CASES INVOLVING FORENSIC DNA ANALYSIS-Key to solving ‘Stolen Trees’ Cases,
-Molecular geneticist Keith Woeste at Purdue University was able to prove that the walnut wood came from a tree stolen in Warren County.
-Woeste ground the wood into a fine powder to extract enough DNA evidence to convince investigators the wood matched that of the stolen tree, which had been sold to a sawmill.
(Purdue.edu, 2004)
RECENT PROGRESS AND ISSUES
RECENT PROGRESSOral Microbiota as a tool to estimate Time Since Death (TSD)
RECENT PROGRESSSex determination based on Raman Spectroscopy of saliva traces for forensic purposes
- Raman spectroscopy is a non-destructive technique that can be used at the crime scene to determine the sex of saliva donor
- Can be used in the absence of DNA Profile or in cases where DNA results does not yield a match in a database - RAMAN SPECTRA
RECENT PROGRESSDNA profiling in crimes involving monozygotic twins
Researchers used ultra-deep generation sequencing associated with bioinformatics techniques in differentiating DNAs of monozygotic twins
Results: 5 Single Nucleotide Polymorphisms (SNPs) were present in the twin father and his child but absent in twin uncle
Conclusion: Rare mutations will occur early after the human blastocyst splits into two (the origin of twins) and that such mutations will be carried on into somatic tissue and germline
RECENT PROGRESSForensic Science Institute Act
Establishing Forensic Science Institute in the University of the Philippines System
Objectives:
● Promote R&D● Enhance existing laboratories● To develop a corps of professionals
CURRENT ISSUES
familial searching in CombinedDNA Index System (CODIS)Database could affect innocentpeople whose DNA would partiallymatch with that found in the crimescene (Gloudemans & Shamaprasad,2015).
CURRENT ISSUES
In the Philippines,● Need for fingerprints and DNA database (i.e. Automated
Fingerprint Identification System or AFIS)● Cases of malpractice in investigating dead bodies, writing
autopsies and dead reports● Lack of support and participation of the legislative branch
BIOTERRORISM
Outline
I. INTRODUCTION
A. BIOTERRORISM
B. HISTORY
C. BIOLOGICAL AGENTS
II. TECHNIQUES AND METHODS USED
III. EXAMPLES
IV. RECENT PROGRESS AND ISSUES
WHAT IS BIOTERRORISM?Bioterrorism is intentional use of microorganisms or toxins derived from living organisms to cause death or disease in humans or the animals and plants on which we depend.
● Catastrophic terrorism with mass casualties● Microevents using low technology causing
civil unrest, disruption, disease, disabilities and death● Spreading a virulent disease among animal production
facilities● Poisoning water, food crops and supplies
Das and Kataria, 2010
NATIONS DISSIDENT GROUPS
HISTORYThe use of biological agents as weapons is NOT a new concept since it is known to be used as early as the ancient times.
Prakash et al., 2010
650 BC Assyrian politicians dumped fungus (Claviceps purpurea) from rye into their opponents′ wells, giving them fatal ergot poisoning.
400 BC Scythian archers infected their arrows by dipping them in decomposing bodies or in blood mixed with manure.
HISTORYThe use of biological agents as weapons is NOT a new concept since it is known to be used as early as the ancient times.
Prakash et al., 2010
300 BC Persian, Greek, and Roman used dead animals to contaminate water sources such as wells.
190 BC Hannibal, a Carthaginian commander, won a naval victory by firing earthen vessels full of venomous snakes into the enemy ships.
HISTORYThe use of biological agents as weapons is NOT a new concept since it is known to be used as early as the ancient times.
Prakash et al., 2010
1346 Tatar armies spread bubonic plague bycatapulting diseased corpses over the walls of the city of Kaffa
1767 Spreading of smallpox via contaminated blankets by the British to the Native American population loyal to the French.
HISTORYThe use of biological agents as weapons is NOT a new concept since it is known to be used as early as the ancient times.
Prakash et al., 2010
WWI US & Germany developed biological weapons using anthrax & glanders.
WWII Japan operated Unit 731 that carried out human experiments on prisoners exposing them to plague, anthrax, syphilis, and other agents.
CHARACTERISTICS OF BIOLOGICAL AGENTS
EFFECTIVE AT LOW DOSE DIFFICULT TO IDENTIFY
AGENTS OF BIOTERRORISM
CLASSIFICATION, ROUTES OF ENTRY & METHODS OF DELIVERY
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY A● Easily disseminated● Causes high mortality● Causes public panic and social disruption● Requires special attention for public health preparedness
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY A● Examples: Bacillus anthracis (anthrax), Clostridium botulinum
(botulism), Yersinia pestis (plague), Variola major virus (smallpox), Francisella tularensis (tularaemia), Filo viruses and Arena viruses (viral hemorrhagic fevers), Ebola virus (Ebola hemorrhagic fever), Marburg virus (Marburg hemorrhagic fever), Lassa virus (Lassa fever), etc.
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY B● Moderately easy to disseminate● Causes moderate morbidity● Requires enhanced disease surveillance and public health
diagnostic capacity
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY B● Examples: Alpha viruses, Eastern, western, and Venezuelan
equine encephalomyelitis viruses (EEE, WEE, VEE), Brucella species (brucellosis), Burkholderia mallei (glanders), Coxiella burnetti (Q fever), foodborne and waterborne pathogens (Cryptosporidium parvum, Escherichia coli O157: H7, Salmonella species, Shigella dysenteriae, Vibrio cholerae, etc.)
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY C● Could be engineered for dissemination in the future● Have potential for high morbidity, mortality, and major health
impacts● Examples: Hanta viruses, Multidrug-resistant tuberculosis,
Nipah virus, Tick-borne encephalitis viruses, Tick-borne hemorrhagic fever viruses, Yellow fever
CLASSIFICATIONAccording to the Centre for Disease Control and Prevention (CDC),currently known biological weapons can be classified into 3categories.
Das and Kataria, 2010
CATEGORY C● Could be engineered for dissemination in the future● Have potential for high morbidity, mortality, and major health
impacts● Examples: Hanta viruses, Multidrug-resistant tuberculosis,
Nipah virus, Tick-borne encephalitis viruses, Tick-borne hemorrhagic fever viruses, Yellow fever
ROUTES OF ENTRY
METHODS OF DELIVERY
● Through contaminated food and water
● Through air-handling systems
● Through bomblets delivered by aircrafts
● Through spray tanks released via aircrafts and other
vehicles or via tall buildings
● Through delivery by post
● Through deliberate infiltration of infected animals, vectors,
and pests
SPECIFIC EXAMPLES
ACTS OF BIOTERRORISM IN THE 20TH-21ST CENTURY
GLANDERS● 1915● Dr. Anton Dilger, developed a microbiology facility in
Washington, D.C. that produces large quantities of anthrax and glanders bacteria
● 3,000 horses, mules, sheeps & cattle shipped from Norway, Spain, Romania, and the United States
● To interrupt the flow of supplies to the Allied frontlines● Inoculated using needles and capillary tubes● Effects are unknown since no reports have been made
of disease outbreaks among livestock.
MYCOTOXIN● 1981● Attacks in Vietnam, Laos, Cambodia & Afghanistan
linked to Soviet Union● Mold byproduct of Fusarium spp.● Helicopter or plane flying over releasing a yellow
colored cloud that would fall in a manner that looked, felt and sounded like rain
● Known as “Yellow Rain”● “Naturally occurring phenomenon of a swarm of Asian
honeybees defecating in flight”
SALMONELLA● 1981● Bhagwan Shree Rajneesh cult● Contaminated salad bars with Salmonella bacteria in a
small town in Oregon● Aim: To influence the local county elections.● Largest scale act of bioterrorism in U.S. history● More than 750 cases of salmonellosis
RICIN● 1991● Ricin - a ribosome-binding protein naturally occurring
in the seeds of the castor oil plant.● Patriots Council, an antigovernment extremist group,
members arrested for plotting to kill a U.S. marshal with ricin
● Smearing it on the marshal’s car door handle● Lethal when inhaled or absorbed through damaged
skin.● After 2-5 days, damage to nervous system and internal
organs
SMALLPOX● 1992● Soviet Union had the ability to launch missiles
containing weapon grade smallpox● Smallpox killed at least 300 million people ● At present, smallpox is one of the biggest threat since
most vaccines are destroyed as per recommendations of WHO advisory committee
ANTHRAX● 1994● Aum Shinrikyo, a Japanese religious sect released
anthrax through aerosol from the tops of buildings in Tokyo
● However the attack was a total failure since the group used the vaccine strain of the bacterium.
ANTHRAX● 2001● Letters containing anthrax
spores were mailed to a television news anchor, US senator, and others
● 5 deaths from pulmonary anthrax and 17 other cases of inhalation and cutaneous anthrax.
RECENT PROGRESS AND ISSUES
IMPLEMENTING LAWS ON BIOTERRORISMGENEVA PROTOCOL● Also known as “Protocol for the
Prohibition of the Use in War of Asphyxiating, Poisonous or other Gases, and of Bacteriological Methods of Warfare.”
● Signed at Geneva on June 17, 1925 ● Entered into force on February 8,
1928● PH gave its depositary on June 8,
1973● Countries that violated the law:
Spain, France, Japan, US, UK, Germany, Soviet Union, Iraq, Syria
IMPLEMENTING LAWS ON BIOTERRORISMBIOLOGICAL WEAPONS CONVENTION (BWC)● Also known as “Convention on the Prohibition of the Development,
Production and Stockpiling of Bacteriological (Biological) and Toxin Weapons and on their Destruction.”
● Signed at London, Moscow, and Washington D.C. on April 10, 1972 ● Entered into force on March 26, 1975● PH gave its signed on April 10, 1972 (L, W) and on June 21, 1972 (M);
and gave its depositary on May 21, 1973 (W) ● Occasionally holds review conferences regarding issues and
implementations of the law
IMPLEMENTING LAWS ON BIOTERRORISMREPUBLIC ACT NO. 9271● Also known as “"Implementing Rules
and Regulations of the Quarantine Act of 2004.” or simply “Quarantine Act”
● Creation of the Bureau of Quarantine ● Monitors aircrafts and transport vessels
for any radiological, biological, and chemical (RBC) weapons
● Also responsible for any RBC incidents that may happen inside the country such as bioterrorism
COMMON REACTIONS OF PEOPLE TO BIOTERRORISM● Vicarious Rehearsals - These are emergency protocols or
courses of action done as a safety precaution of people
unlikely to be affected by the bioterrorism attack.
● Denial - This is a reaction wherein people refuse to
acknowledge the existence of a threat or to take necessary
action towards it.
● Stigmatization - This is the fear and isolation of a group
perceived to be contaminated or risky to associate with.
● Fear and Avoidance
● Withdrawal, Hopelessness, and Helplessness
DETECTION AND PREPARATION FOR BIOTERRORISM ATTACKSIntelligence and Information Management
● People’s awareness to biological weapons and other bioterrorism agents
● Scientists’ further research on the matter and help explain in layman’s
terms the bioterrorism agents
● Exchange of information between intelligence, S&T, and public health
communities
BioWatch - “BioWatch air monitoring and analysis, notification procedures, and risk assessment can eliminate or substantially minimize the catastrophic impact of a biological attack. BioWatch also gives public health, law enforcement, emergency management, and laboratory communities a forum to help develop a common understanding of a bioterrorism incident, focus resources, and determine the appropriate response.”
DETECTION AND PREPARATION FOR BIOTERRORISM ATTACKSIdentification of Biological Agents in Environment
● Traditional laboratory detection is slow and limited. However, it is
also the most reliable approach especially in developing countries.
● Develop and evaluate rapid, sensitive, and specific early-detection
technologies.
DETECTION AND PREPARATION FOR BIOTERRORISM ATTACKSSurveillance and Diagnosis of Infection and Disease
● Complications due to lack of relevant medical experience in
handling bioterrorism agents in addition to nonspecific symptoms
of the related disease (fever, flu, etc.)
● Surveillance and diagnosis should focus on three areas: preclinical,
human disease, and agricultural.
ARE WEPREPARED?
BIOTERRORISM IN THE PHILIPPINES
“The Philippines is ill-equipped to deal with a chemical or
biological attack by terrorists because its cash-strapped police
have neither the equipment nor the technology to handle
these "dirty weapons" of mass destruction.”
Singapore Institute of International Affairs (SIIA), 2005
BIOTERRORISM IN THE PHILIPPINESAccording to Maria Auxilia T. Siringan, the head of the
Microbiological Research and Services Laboratory, University
of the Philippines Diliman, Quezon City, the Philippines is:
• Not capable of detecting or stopping the spread of
bioterrorism attacks
• Still in the conceptual stage of creating a biodefense system
prepared to stop the intentional release of biological agents
or toxins such as plague, smallpox, anthrax or the Ebola
virus.BioPrepWatch Reports, 2012
BIOTERRORISM IN THE PHILIPPINES
• Last November 7, 2016 at the 8th Review Conference of the
Biological Weapons Convention in Geneva, Switzerland, the
Philippines, as one of the state parties, highlighted its efforts
in raising awareness in bioterrorism by implementing the
National Chemical Biological Radiological Nuclear (CBRN)
Action Plan and its active engagement in counter-terrorism
measures within the Asia Pacific Region.
DFA, 2016