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Objectives - Morning Session

Introduction to Biology
Compare Prokaryotic and Eukaryotic cells and list structures common to
both types of cells
Describe the function of cells and DNA
Describe the basic differences between bacteria, viruses and toxins

Bioterrorism & Select Agent Overview

Define Bioterrorism; familiarize with history aspect of BT
List agents on the A, B & C lists, explain why they are on those specific
lists
Learn about biological agents that are on the Select Agent A List

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 Objectives - Afternoon Session
Clandestine Labs & Agent Detection
Recognize the potential indicators of a biological threat (clan labs)
Explain the different types of presumptive biological detection testing
technology; recognize their limitations, and the importance of sending all
samples to the appropriate lab for definitive testing and identification

Sampling and Packaging

Describe forensic sampling protocols which are currently in place in
Canada
Identify the different sampling tools required to retrieve various samples
Use the sampling protocols to collect samples for laboratory analysis

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Infectious Disease
Through the ages:-

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SMALLPOX

Signs of Smallpox

Electron micrograph of a
1157 B.C Ramses V, an Egyptian Pharaoh Smallpox virus 5
PLAGUE

Sign of bubonic plague

Plague also known as the Black

Death
Yersinia pestis, the bacterium that
causes Plague 6
PLAGUE ROUTE

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 1918 Spanish INFLUENZA

Influenza virus
showing surface antigens

Geographic origin unknown

Victims healthy young adults
Spread was facilitated via close
troop quarters and massive troop
movements Electron micrograph of
influenza virus 8
Biological Basics
 Cells

All living things are made up of cells

There are two types of cell:
Prokaryotic cells e.g Bacteria
Eukaryotic cells e.g Plant and animal cells

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Prokaryotic cells

www.cellsalive.com
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Eukayrotic cells
Nucleus
Cytoplasm

Nuclear
Cell
membrane
membrane

Ribosomes

Golgi bodies
Mitochondria
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Prokaryotic vs Eukaryotic

Prokaryotic means Eukaryotic means true

without nucleus nucleus
All bacteria are All plant and animal
prokaryotic cells. cells are eukaryotic
Simple structure cells.
Very small- diameter Complex structure
ranges from between 1 10 times larger than
10 micrometers (m) prokaryotic cells
diameter ranges from
between 10 100
micrometers (m)
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Cell structure
Some structures are found only in
prokaryotic cells
Nucleoid
Cell wall
Some structures are found only in
eukaryotic cells
Nucleus
Mitochondria
Nuclear membrane
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Some structures are common to both

types of cells
Cytoplasm
Cell membrane
Ribosomes
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Cell function

The main job of a cell is to make the

molecules that it needs to survive, grow,
multiply and do its job.

The molecules that all cells make are

called proteins.
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Cells use DNA to make proteins

DNA (deoxyribonucleic acid) is inside

every living cell
DNA directs vital activities such as
growth, division, movement and even
death
DNA provides the instructions cells use to
build proteins
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The chemical language for

this code is very simple.
It consists of four molecules
called nucleotides.
They are Adenine, Thymine,
Guanine and Cytosine

EVERY ORGANISM & INDIVIDUAL HAS A

UNIQUE DNA SIGNATURE
 Video
Every Creature has DNA

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 1 22
Nucleus- is the control centre of all
cells, it holds all the instructions
needed to make proteins.

DNA Recipe
book
Locked inside
the nucleus

2 3
RNA makes a copy of the
instructions written in the mRNA takes these
recipe book instructions to the ribosome
inside the cell cytoplasm

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Ribosome- is like a chef. It follows the
instructions delivered by the mRNA to
put amino acids together in the right
order to build proteins

Proteins- are complex molecules that do most

5 of the work in our bodies. Once proteins are
put together they are transported to where
they are needed. Proteins may be used inside
the cell, sent out to its surface or excreted from
the cell entirely. Antibodies and hormones are
examples of proteins.
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Video
What happens within the cell
& how proteins are made
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Importance of DNA & RNA

Every living thing on this planet has UNIQUE
DNA signature
All Biological agents - Bacteria, Viruses and
Toxins have unique DNA signatures
Scientist use these unique DNA signatures to
identify specific biological agents
How do we get to DNA?....
 By a method called DNA isolation
Routine procedure used to collect DNA for molecular and forensic
analysis with 3 steps:
BreakingthecellsopentotheexposetheDNA
(bychemicalorphysicalmethods)
Removinglipidsbyaddingdetergent
PrecipitatingtheDNAwithanalcohol
DNAnormallystaysdissolvedinwater,but
whensaltyDNAcomesincontactwith
alcoholitbecomesundissolved
(precipitation).Thephysicalforceofthe
DNAclumpingtogetherasitprecipitates
pullsmorestrandsalongwithitasitrises
intothealcohol

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 FYI only
DNA is a long, stringy molecule. The salt that you added in step one helps it
stick together. So what you see are clumps of tangled DNA molecules!
You can use a wooden stick or a straw to collect the DNA. If you want to
save your DNA, you can transfer it to a small container filled with alcohol.
ethanol or isopropanol. Since DNA is insoluble in these alcohols, it will
aggregate together, giving a pellet upon centrifugation. This step also
removes alcohol-soluble salt.
Cells open, commonly referred to as cell disruption or cell lysis, to
expose the DNA within. This is commonly achieved by chemical and
physical methods-blending, grinding or sonicating the sample.
Removing membrane lipids by adding a detergent or surfactants.
Removing proteins by adding a protease (optional but almost always done).
Removing RNA by adding an RNase (often done).
Precipitating the DNA with an alcohol usually ice-cold ethanol or
isopropanol. Since DNA is insoluble in these alcohols, it will aggregate
together, giving a pellet upon centrifugation. This step also removes alcohol-
soluble salt.
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Importance of DNA
Scientists use to extract DNA from many
different sources
It is usually the first step in many
laboratory experiments
DNA extraction can be used to create DNA
fingerprints to help
diagnose genetic diseases
solve criminal cases
identify biological agents!!
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Biological agents
Bacteria, viruses and toxins are biological
agents that can cause disease in plants,
animals or humans
Including by intentional means!!
Unlike nuclear and chemical agents,
biological agents do not give off any
signatures which are detectable with
currently available hand-held devices
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Biological agents
Undetectable by senses
More toxic than chemicals by weight
Signs and symptoms are delayed
Unique DNA signatures and proteins
enable biological agent detection
More difficult to identify that chemical agents
Limited field detection
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Biological agents
Bacteria

Infectious capable of causing infection

Contagious can spread from person to person
no nucleus
DNA is circular and is not organized into chromosomes
no membrane-bound organelles in the cytoplasm
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Biological agents
Viruses

Infectious capable of causing infection

Contagious - can spread from person to person
Simple, acellular organization
Contains DNA or RNA (not both) enclosed in a coat of
protein
Cannot reproduce independently of living cells
Use host metabolism to replicate and reproduce
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Biological agents

Toxins

Not infectious Not capable of causing an infection

Not contagious - cannot spread from person to person
Poisonous substances produced by animals, plants or
bacteria
Usually proteins, ex. Ricin, Botulinum, SEB
 Sizes Of Biological Agents
(smallest to largest)

Viruses - 0.02 to 0.3 microns

Bacteria - 0.5 to 10 microns
Toxins - less than one millionth
the size of a bacteria

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Relative sizes
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*PIN 1x *
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*PIN 1x *
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2x
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4x
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6x
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8x
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*10x *
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HAIR 20x
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40x
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DUST MITE 60x
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80x
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*100x *
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200x
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400x
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600x
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RAGWEED POLLEN 800x
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* 1000x *
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LYMPHOCYTE 2000x
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RED BLOOD CELL 4000x
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BAKERS YEAST 6000x
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*10,000 X *
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STAPHYLOCCOUS 20,000 X
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40,000 X
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60,000 X
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EBOLA 80,000 X
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*100,000 X*
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200,000 X
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400,000 X
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RHINOVIRUS 600,000 X
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800,000 X
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*1,000,000 X*
 Bacteria:- Closer Look
Single celled organism
containing DNA
Do not need a host to
survive
Alive
M ove
R espire
S ensitive
G row
R eproduce
E xcete
N utrients
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 Bacteria

Some can form

spores
Resistant to harsh
environments
ANTHRAX
BOTULINUM

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Bacterial Reproduction

Reproduce by
asexual process

Binary fission is
where a single cell
divides into two
identical cells
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Video
Binary Fission
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Viruses
Are NOT cells
Contain either RNA or DNA surrounded by a
protein coat
Are NOT alive
Do not move, grow, respire
Do not have a nucleus, cytoplasm or membrane
Need a host to survive, must hijack cells
Can only reproduce in living host cells
 Are viruses alive?

Only 1 characteristic of life:

Reproduction
Can only reproduce inside a host cell!
Process or reproduction = Lytic cycle

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Viral reproduction

Lytic cycle - this

form of viral
replication kills the
host cell
Virus attaches to host cells
membrane and injects its
nucleic acid into the host cell.
The viral nucleic acid takes
over protein synthesis, creating
new viruses.
The host cell bursts, lyses,
releasing the newly formed
viruses
 Viruses
Viruses are host specific
A protein on the surface of the virus has a shape
that matches a molecule in the plasma membrane of
its host, allowing the virus to lock onto the host cell

Viruses are prone to genetic information

exchange
Mutation
Recombination
Reassortment
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 Video
How we get different viral
combinations

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Reassortment Events: 2009
Influenza A (H1N1) Virus

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 In summary

Viruses are non-living

They are much smaller then the cells they
enter
Can only reproduce with the help of host
cells
Prone to genetic information exchange

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Growing bacteria and viruses in a

laboratory

Bacteria Viruses
Need nutrients and Need a host cell in order
warmth to grow to replicate
Grown on agar plates, Grown in tissue culture
which supply nutrients tissue culture provides
Incubated at a suitable viruses with cells that they
temperature. can invade and hijack.
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Growing bacteria and viruses

Specialised equipment is required

depending on what you are growing

This equipment can help you determine

what a terrorist might be growing in a
lab
 Anthrax Plague Tularemia

Nutrient
broth

Bacteriallabequipment
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Viral indicators Viral incubator

ViralLab
Equipment
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Microscopes

Light microscope Electron microscope

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Electron micrographs of Viruses

SARS Ebolavirus
coronavirus

Influenza Herpes virus

virus
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Toxins

Are NOT cells

Poisonous substances, usually proteins,
produced by animals, plants or bacteria
Clostridium botulinum makes botulinum toxin
Staphylococcus aureus makes Staphylococcus enterotoxin B
(SEB)
Seeds from a castor bean plant make ricin
 Toxins
Small proteins/peptides
Metabolic by-products:-
Bacterial toxins
cholera, botulinum, diphtheria etc.
Plant toxins
Ricin, abrin etc.
Animal/Insect toxins
Venom, deep sea fish, etc.
Molds, Yeast, Fungi
Sick Building Syndrome
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 In summary.
BACTERIA VIRUSES TOXINS

Infectious? YES YES NO

Able to YES YES only in a NO

Replicate? host cell
Has Genetic YES YES NO it is a
Material (DNA)? protein
Survival in the Hours to Years* Hours to days Hours to Years
environment (spores)
Questions?