Biology

Laboratory Safety
Biohazard Safety Module

Northern Michigan University

Biology Department

This presentation is designed as a training

course for those students who will be intentionally
working with Biohazardous materials
Biohazards are defined as those materials or
organisms that carry the possibility of causing
disease

The NMU Institutional Biological Safety

Committee oversees research and teaching that
deals with
Infectious Agents (includes attenuated lab &
vaccine strains)
Biologically Derived Toxins
Recombinant DNA
Artificial Gene Transfer

Biohazardous Agents by Risk Groups (RG)

RG1

Agents that are not associated with disease in healthy adult humans

RG2

Agents associated with human disease that is rarely serious or for

which preventive or therapeutic interventions are often available

RG3

Agents that are associated with serious or lethal human disease for
which preventive or therapeutic interventions may be available.
The agent is of low risk for the community

RG4

Agents that are likely to cause serious or lethal human disease for which
preventive or therapeutic interventions are not usually available. The
agent is of low risk for the community.

Agents: any infectious entity or any molecule produced by them capable of

producing disease in humans

At NMU, we are approved to work with RG1 and RG2 agents for
research purposes. However, students could be exposed to RG3 agents
from the environment.

Selected agents

Examples of pathogens and toxins

RG1 group
E. coli nonpathogenic laboratory strains
Saccharomyces cereviseae
Bacteriophages

RG2 group
E. coli, pathogenic strains
Influenza virus
Pseudomonas aeruginosa

Examples of pathogens and toxins

RG3 group
Human immunodeficiency virus
Histoplasma capsulatum
Mycobacterium tuberculosis

RG4 group
Ebola virus
Marburg virus

Routes of transmission of infectious agents in

the lab setting:
Oral Ingestion

Eating, drinking, smoking

Broken skin

Needle-stick
Cuts
Scratches
Bites

Mucosa

Splash of liquids
Handling eye contact lenses
Applying make-up

Upper respiratory tract Inhalation of aerosols

Oral Ingestion

NO drinking or eating in the lab!

It can take fewer than 10 E. coli O157 H7 to infect a human!

International Biohazard Symbol

Universal Symbol
Communicates potential exposure
Red or orange

Any container with biohazard potential must be

labeled with this symbol

Broken skin
Take care when dealing with
SHARPS

Needles and syringes, scalpels, blades, broken

glass, etc.
Glassware exposed to an infectious agent such as
Pasteur pipettes, glass containers, etc.
Any potentially contaminated sharp should be disposed
of in a Biohazard Sharps container

Sharp Containers for Biohazards

Sharp containers

are red,
display the International Biohazard Symbol
are labeled Medical Waste or Infectious Waste
are puncture proof

Mucosa Exposure Routes

No!

Good safety practice!!

Make-up

Contact lenses

Upper respiratory tract exposure methods

Some activities that generate aerosols take care!
Opening tubes
Centrifugation
Flaming to sterilize tools
Vortexing
Pipetting
Electroporation
Sonication
Flow cytometry
REMEMBER: An agent that is not normally airborne transmitted can
be transmitted in the lab by aerosols

Other Potential Sources of Infection

Blood and Other Potentially Infectious Material (OPIM)
Follow "Universal Precautions"
all human blood and certain human body fluids are
treated as if known to be infectious

Environmental Samples:
soil, plants, water samples, animals, etc.
Follow NMU Precautions
all environmental samples are treated as potential
health hazards

Containment of Biohazard Agents

The purpose of containment is to reduce or eliminate exposure
of laboratory workers & the outside environment to hazardous
agents
Primary containment: protection of people & immediate
laboratory environment
It is achieved by the use of
Good Microbiological Techniques
Safety Equipment
Secondary containment : protection of external environment
It is achieved by the use of
Facility Design
Operational Practices

Primary containment
Protection of people and the immediate laboratory environment

Good Microbiological Techniques

Use proper Safety Equipment (Primary Barriers):
Personal Protection Equipment (PPE): gloves,
lab coats, goggles, face shields, etc.
Engineering controls designed to remove or minimize
exposures to hazardous biological materials:
safety centrifuge cups, biological safety cabinets, etc.

Safety Equipment

Biological Safety Cabinets (BSCs) type II

BSCs type II: protect workers when used in combination with

good microbiological techniques, &
prevents external contamination by
materials being manipulated inside cabinet

Proper use of Biological Safety Cabinets (BSCs)

Maintain constant air curtain
Minimize movements in/out
No Burners inside the BSC
Disinfect working surface and interior
Avoid clutter, keep grille clean
UV lamps will be used with care and only for 30 min.
Do not use volatile chemicals inside the BSC
Certify performance annually (We annually certify our BSC at NMU)

Secondary containment
Protection of external environment

Facility Design
Contributes to the protection of people working in labs
Provides a barrier to protect persons outside the labs
Protects persons or animals in the community
Operational Practices
Determined by Risk Group of biohazard agents

RISK GROUP 1
Agents not associated with disease in healthy adult humans

BioSafety Level 1 (BSL-1)

Primary Barriers

No safety equipment required

Secondary Barriers Open bench and sink

Good Microbiological Practices for BSL-1

Limited laboratory access unless responsible permits

No eating, drinking, applying makeup, etc.
No mouth pipetting
Safe handling of sharps
Safety glasses
Lab coats (must stay in the lab until washed)
Hand washing using antimicrobial soap
Decontaminate spills, cultures and waste

CDC guidelines for BSL 1 labs

Access to the laboratory is limited or restricted at the discretion of the
laboratory director when experiments or work with cultures or specimens is in
progress.

Hand washing for Safety

Handwashing with soap and water remains a sensible strategy
for hand hygiene in non-health care settings
Hand Hygiene Guidelines Fact Sheet , CDC

Method:
Twenty seconds of vigorous washing with soap and
water

When:
After lab section or as often as needed

Disinfection Practices

When?
Before and after lab experiment
To decontaminate spills
How?
Add disinfectant and wipe with a paper towel
In the case of a spill, apply disinfectant around the spill
and wipe with the paper towel from the periphery to the
center
Discard paper towels in the biohazard bag

RISK GROUP 2
Agents associated with human disease that is rarely serious
or for which preventive or therapeutic interventions are often
available

BioSafety Level 2 (BSL-2)

Good Laboratory Practices of BSL-1 plus
Limited access
Biohazard warning signs
'Sharps' precautions
Biosafety manual defining any needed
waste decontamination

Procedures for disposal of Biohazards

Into biohazard bags:
All cultures and biological fluids
All contaminated glass
All needles & syringes
All other potential biohazard material
Filled biohazard bags are autoclaved for 40 min. at
121oC & 15 lbs/in2
After biohazard bags and cardboards are
autoclaved, they are bagged in black plastic
bags

Autoclave Guidelines
Equipment must continuously monitor & record temperature &
pressure during entire length of each cycle.
Temperature-sensitive tape must be affixed to each bag or
container.
Effectiveness must be evaluated under a full load at least once
every 40 hours of operation
Each bag must be exposed to a minimum of:
121 degrees Celsius
15 pounds of pressure per square inch
At least 30 minutes time AT THESE CONDITIONS

Autoclave Recordkeeping
A written log shall be maintained for each unit & shall
contain the following:
Date, time, duration, & operator of each cycle
Approximate weight/volume of medical waste treated during
each cycle
Temperature & pressure maintained during each cycle
Method utilized for confirmation of temperature & pressure
Dates & results of calibration & maintenance

Written log record must be retained for three years

Right to Know Policy

You always have the right to ask questions, or
report hazards, either directly or anonymously
without any fear of reprisal to
Your lab instructor and/or the course instructor
Biology Department Head and Chemical Hygiene
Officer Dr. John Rebers (jrebers@nmu.edu)
Public Safety (227-2151)

Next
Please complete the PRESENTATION
REVIEW ACKNOWLEDGEMENT
Then complete the QUIZ that follows the
confirmation
You must achieve a perfect score on the
quiz to successfully complete your safety
training. You may retake the quiz as many
times as necessary.
Failure to complete your safety training
will result in your exclusion from the
laboratory portion of your course

After the quiz

Remember to complete any additional Safety
Training modules that are required for your
course.