Transcript File

Basics of Biosafety
Working Safely with Biological Materials
What is Biosafety?

Principles and practices employed to protect
laboratory personnel and the environment from
exposure or infection while working with living
organisms, biological materials, or agents.
 Included
are any materials that may be potentially
infectious.
 Includes recombinant DNA research
Agents and Risks

The “agent” is the what creates risk

Risks to the worker or environment are
often unknown

Determining “acceptable risk”?
Assessing Risk
 There
The
is always risk!
risk must be identified
The risk is evaluated
The risk must be measured
Plan to minimize the risk
Who Determines Acceptable Risk?
Assessment is conducted by a Biosafety
Professional in partnership with and based
on information provided by the Principal
Investigator
 The assessment is presented to the
Institutional Biosafety Committee (IBC) for
approval

Identifying Risk
Understand the biology of the agent
 Susceptibility and transmission within the
host
 Hazards associated with equipment and
procedures
 Goal:

 Provide
the highest practical protection and
the lowest practical exposure
Evaluating Risk Acceptability

Worst case scenario -What might happen?

Likelihood of an event

Seriousness of the incident

Actions needed to resolve the problems
What is Acceptable Risk?

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
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
Since there is no such thing as “no risk”
“Safe” means risk has been judged acceptable
Judging risk is a subjective- humans make
decisions
Measuring risk is objective- use available
guidelines, data, and documentation
Keep records of how determinations were made
due to subjective nature of the process
WHO-World Health Organization
Agents Assigned Risk Groups

RG-1 Unlikely to cause disease in humans or animals

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RG-2 May cause disease but typically not serious

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individual risk, low community risk, treatable
RG-3 May cause serious disease, usually treatable

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low individual or community risk
High individual but low community risk, serious respiratory agents
RG-4 Serious or fatal, often not treatable,

Easy transmission, high individual and community risk
Biosafety Levels (BSL)

Different than the Risk Groups!!
 Risk
groups used in risk assessment
 BSL are used in risk management

BSL are ways to control the agent
 facilities,

safety equipment, practices, PPE, etc.
Once risk is assessed then the appropriate BSL
is determined
BioSafety Level 1

Well characterized, non-pathogenic
organisms or agents

Open bench- no containment

Use good laboratory practices, waste
disposal, and aseptic techniques

Example: E. coli K-12 strains
BioSafety Level 2

Agents of moderate hazard to personnel
or environment

Basic lab, but restricted access, containment during
certain processes (i.e. aerosols, large volumes, etc.)

Autoclave and Biological Safety Cabinet desired

Use good laboratory practices, waste disposal, and
aseptic techniques

Example: most non-respiratory, non lethal, agents
BioSafety Level 3

Agents of high hazard to personnel or environment

Respiratory exotic or indigenous agents which are
easily transmissible causing serious or lethal disease

All work is contained, engineering controls and
controlled environments we currently do not have the
facilities to handle.
Example: Mycobacterium tuberculosis, SARS, etc.
BioSafety Level 4

FORGET ABOUT IT!!!

Hemorrhagic fever, deadly viruses, etc.

Total containment, airtight labs, “submarine”
doors, air pumps, water treatment, HEPA
filtration, etc.

Positive pressure “moonsuits”
Laboratory Acquired Infections (LAI)
Bacterial:
76% from clinical labs
8% from research labs
Exposure:
60% acquired from inhalation
Other exposures include:
digestion, sharps, splashes, direct and indirect contact
Laboratory Acquired Infections (LAI)
Viral
16% from clinical labs
 70% from research labs

 32%
from animal related activities
Biohazardous/Medical Waste
Waste
that is potentially infectious to
humans, animals or plants. It includes:
 Medical
Waste according to MMWRA
 Regulated Waste by MIOSHA
 Regulated Waste by CDC/NIH
Michigan Medical Waste Regulatory Act
(MMWRA)
Defines “medical waste”
 Requirements for waste handling and
disposal
 Requires generators to register with
DEQ and implement a Medical Waste
Management Plan

Michigan Medical Waste Regulatory Act
(MMWRA)
Defines “medical waste”
 Requirements for waste handling and
disposal
 Requires generators to register with
DEQ and implement a Medical Waste
Management Plan

Biohazardous Waste Management Plan
Must outline how generating facility
complies with the MMWRA:
 Types of wastes generated
 Storage and disposal of wastes
 Contingency plans
 Training
Biohazardous Waste Categories
Cultures and stocks of infectious
agents and associated biologicals
 laboratory
waste
 biological production waste
 discarded live and attenuated vaccines
 culture dishes and related materials
 contaminated PPE
Biohazardous Waste Categories
Liquid human and animal waste
 liquid
or semi-liquid blood and blood
products and body fluids
 contaminated
items that would release
blood or items that are caked with blood
or other potentially infectious materials;
NOT including urine or materials stained
with blood or body fluids
 infectious
animal waste (research)
Biohazardous Waste Categories
Pathological waste
 tissues
 body
parts other than teeth
 products of conception
 fluids removed by trauma or during
surgery or autopsy/necropsy or other
medical procedure and not chemically
fixed.
…And More Biohazardous
Waste Categories
Animal and plant pathogen waste
 Recombinant DNA waste
 Sharps

Biowaste vs. Trash
3 basic questions to differentiate:
1. Is it contaminated with viable
biological material?
2. Can blood or other regulated
body or biological fluids be
released?
3. Is it a sharps hazard?
#1
Is it contaminated with
viable biological
material?
Examples:
• Contaminated lab waste
• Personal protective equipment
used for handling potentially
infectious materials (including
handling infected animals or their
products)
• Wastes from infectious disease
research (carcasses, body
fluids…)
#2
Can blood or other
(regulated) body fluids or
viable biological materials be
released?
Some Examples…
Tubes of blood
Vacuum flasks containing body
fluids or cell line waste
Managing Liquid Biohazardous Waste
Storage:
 Label and secure bulk vessels
if not disposed of immediately
Treatment:
 Chemical disinfection OR
 Autoclave
Disposal: THEN
 Flush to sewer
 Use proper PPE!
Disinfection

10% bleach solution
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good for general disinfection
High organics use 20%
Needs to be made weekly
Test contact time
Ethanol
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Use 70% solution (most effective)
Longer contact time and flammable
*Should research and know effectiveness and contact time for the
best disinfectant against your agent!
WRAPPERS/NONABSORBENT MATERIALS
CONTAMINATED WITH
BLOOD
BANDAGES/OTHER
ABSORBENTS
SATURATED OR CRUSTED
WITH BLOOD
STAINED?….
or SATURATED?
Managing Non-Sharp Biohazardous Waste

labeled
container
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lined with a
biohazardous
waste bag

equipped with a
lid.
Managing Non-Sharp
Biohazardous Waste
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Securely tie bags for
transport to
treatment/collection
site.
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When moving wastes,
use secondary
containment; avoid
using public halls and
elevators.
“Breakable” Non-sharps Biowaste
Store in labeled containers
that are puncture-resistant,
closable and will capture
leakage, BUT….
…Do NOT use
SHARPS containers!
Effective Waste Autoclaving
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Leave bag open during
autoclaving or loosely closed
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Add water to bag prior to
autoclaving if primarily dry
materials
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Steam must contact
materials
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Place bag in autoclavable
tray with sides
Treated Waste Bag Disposal

Allow waste bag to cool

Use fume hood to reduce
odors

Securely tie bag shut

Place bag in a nontransparent black bag for
regular disposal
Remember: NO ORANGE
BAGS IN DUMPSTER!
#3
Is it a sharps hazard?
Examples:
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needles
syringes
scalpels
all biologically contaminated objects that
can easily penetrate skin (Pasteur pipettes,
razor blades, etc.)
Place sharps in approved sharps container
for disposal!
…Syringes in research settings should
be disposed of as a sharp to avoid
public relations concerns!
Sharps Containers

Containers must be leak-proof,
puncture-resistant, closable & labeled
with the biohazard symbol.

Proper sharps containers
must be used for
both clinic and
field work.
Proper Use of Sharps Containers

Place tops on containers before use
on lab bench

Don’t forget to date the container
when first put into use

Remember: sharps
containers are a
one-way disposal
system
Proper Use of Sharps Containers
Use sharps containers for sharps
ONLY!
•
No solid biohazardous waste (i.e.
gauze, un-broken pipettes, gloves)
•
No mercury
thermometers
What’s wrong with this picture?
Sharps Container Disposal

Containers must be permanently closed
and disposed of through the animal facility
manager:
 Within
90 days
of first use
 When

¾ full
Disposal methods:
 Landfill
 Incineration
 We
use waste hauler
Safety Notes on Sharps Use

Do not re-cap sharps
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Keep sharps container in
close proximity to point of
use (i.e. limit handling) for
easy disposal

Do not leave needles in
pockets of coveralls or
smocks
Carcasses and Body Parts

Human tissues
 Unfixed
tissues are medical waste
 Make waste unrecognizable!

Animal tissues, carcasses
 When
generated in infectious disease or recombinant
DNA research, these are medical waste

These items must be stored in biolabeled,
leakproof containers for incineration.

Waste service- see Audrey Brown
Managing All That Other Waste…
Drain bottles of non-hazardous
materials before disposal in trash
 <3% of volume is considered empty
 Higher volumes must not be thrown
in the trash

Managing All That Other Waste…
Do NOT discard
medications in the
trash.
Return to source for
disposal or seek
assistance from
your campus waste
group.
See Jaime Stock!