Safety and Laboratory

Guidelines
Tarig Mahmoud Hassan
[email protected]
Sources of risk
1- Chemicals
result in injury
2- Equipment

3- Microbial cultures result in infection

Safe handling and disposal of
microbes
Microorganisms present varying degrees of risk to:
 Laboratory personnel (students, technicians, and
faculty)
 People outside the laboratory.
 The environment.
Microbial cultures must be handled safely.
Four biosafety levels (BSLs) that provide a set of
minimum standards for laboratory practices, facilities,
and equipment to be used when handling organisms.
Biosafety levels (BSLs)
1- BSL-1:
Organisms do not typically cause disease in healthy
individuals and present a minimal threat to the
environment and lab personnel.
Standard microbiological practices are adequate.
These microbes may be handled in the open, and no
special containment equipment is required.
Examples include Bacillus subtilis, Escherichia coli,
Rhodospirillum rubrum, and Lactobcillus acidophilus.
Biosafety levels (BSLs)
2- BSL-2:
Organisms are commonly encountered in the community
and present a moderate environmental and/or health
hazard.
These organisms are associated with human diseases of
varying severity.
Individuals may do laboratory work that is not especially
prone to splashes or aerosol generation, using standard
microbiological practices.
Examples include Salmonella, Staphylococcus aureus,
Clostridium dificile, and Borrelia burgdorferi.
Biosafety levels (BSLs)
3- BSL-3:
Organisms are of local or exotic origin and are associated
with respiratory transmission and serious or lethal diseases.
Special ventilation systems are used to prevent aerosol
transmission out of the laboratory, and access to the lab is
restricted.
Specially trained personnel handle microbes in a Class I or
II biological safety cabinet (BSC), not on the open bench.
Examples include Bacillus anthracis, Mycobacterium
tuberculolosis, and West Nile virus.
Biosafety levels (BSLs)
4- BSL-4:
Organisms have a great potential for lethal infection.
Inhalation of infectious aerosols, exposure to infectious droplets, and
autoinoculation are of primary concern.
The lab is isolated from other facilities, and access is strictly controlled.
Ventilation and waste management are under rigid control to prevent
release of the microbial agents to the environment.
Specially trained personnel perform transfers in Class III BSCs. Class II
BSCs may be used as long as personnel wear positive pressure, one-
piece body suits with a life-support system.
Examples include agents causing hemorrhagic diseases, such as Ebola
virus, Marburg virus, and Lassa fever.
Summary of Recommended Biosafety Levels for Infectious Agents
Student Conduct
To reduce the risk of infection, do not smoke, eat, drink, or
bring food or drinks into the laboratory room.
Do not apply cosmetics or handle contact lenses in the
laboratory.
Wash your hands thoroughly with soap and water before
working in the lab, after handling living microbes, and before
leaving the laboratory at any time.
Also, wash your hands after removing gloves.
Do not remove any organisms or chemicals from the laboratory.
Lab time is precious, so come to lab prepared for that day’s
work.
Basic Laboratory Safety
Wear protective clothing (i.e., a lab coat) in the
laboratory when handling microbes. Remove the coat
prior to leaving the lab and autoclave it regularly
Do not wear sandals or open-toed shoes in the laboratory.
Wear eye protection whenever you are heating chemicals,
even if you wear glasses or contacts.
Turn off your Bunsen burner when it is not in use.
Tie back long hair, as it is a potential source of
contamination as well as a likely target for fire.
If you are feeling ill, go home.
Basic Laboratory Safety
If it is your lab’s practice to wear disposable gloves while
handling microorganisms, be sure to remove them each
time you leave the laboratory. The proper method for
removal is with the thumb under the cuff of the other
hand’s glove and turning it inside out without snapping
it. Gloves should then be disposed of in the container
for contaminated materials. Then, wash your hands.
Wear disposable gloves while staining microbes and
handling blood products—plasma, serum, antiserum,
or whole blood.
Basic Laboratory Safety
Use an antiseptic (e.g., Betadine) on your skin if it is
exposed to a spill containing microorganisms.
Never pipette by mouth. Always use mechanical pipettors.
Dispose of broken glass or any other item that could
puncture an autoclave bag in an appropriate “sharps” or
broken-glass container.
Use a fume hood to perform any work involving highly
volatile chemicals or stains that need to be heated.
Find the first-aid kit, and make a mental note of its
location.
Basic Laboratory Safety
Find the fire blanket, shower, and fire extinguisher,
note their locations, and develop a plan for how to
access them in an emergency.
Find the eye wash basin, learn how to operate it, and
remember its location.
Reducing Contamination of Self, Others,
Cultures, and the Environment
Wipe the desktop with a disinfectant (e.g., Amphyl® or
10% chlorine bleach) before and after each lab period.
Never assume that the class before you disinfected the
work area. An appropriate disinfectant will be supplied.
Allow the disinfectant to evaporate; do not wipe it dry.
Never lay down culture tubes on the table; they always
should remain upright in a tube holder.
Even solid media tubes contain moisture or
condensation that may leak out and contaminate
everything it contacts.
Reducing Contamination of Self, Others,
Cultures, and the Environment
Cover any culture spills with paper towels. Soak the towels
immediately with disinfectant, and allow them to stand for 20
minutes. Report the spill to your instructor.
When you are finished, place the towels in the container
designated for autoclaving.
Place all nonessential books and papers under the desk.
A cluttered lab table is an invitation for an accident that may
contaminate your expensive school supplies.
When pipetting microbial cultures, place a disinfectant-soaked
towel on the work area. This reduces contamination and possible
aerosols if a drop escapes from the pipette and hits the tabletop.
Sharps Container: Needles, glass, and other contaminated items that can
penetrate the skin or an autoclave bag should be disposed of in a sharps
container. Do not fill above the dashed black line. Notice the autoclave tape in
the lower left. The white stripes will turn black after proper autoclaving. Above
the autoclave tape is the address of the institution that produced the
biohazardous waste.
Sharps containers
Disposing of Contaminated Materials
In most instances, the preferred method of
decontaminating microbiological waste and reusable
equipment is the autoclave.
Disposing of Contaminated Materials
Remove all labels from tube cultures and other contaminated
reusable items and place them in the designated autoclave
container. This will likely be an open autoclave pan to enable
cleaning the tubes and other items following sterilization.
Dispose of plate cultures (if plastic Petri dishes are used) and
other contaminated non-sharp disposable items in the
designated autoclave container . Petri dishes should be taped
closed. (Note: To avoid recontamination of sterilized culture
media and other items, autoclave containers are designed to
be permanently closed, autoclaved, and discarded. Therefore,
do not place reusable and non-reusable items in the same
container.)
Autoclave bag (The white stripes will turn black after proper autoclaving
Disposing of Contaminated Materials
Dispose of all blood product samples and disposable gloves in the
container designated for autoclaving.
Place used microscope slides of bacteria in a “sharps” container
designated for autoclaving, or soak them in disinfectant solution
for at least 30 minutes before cleaning or discarding them. Follow
your laboratory guidelines for disposing of glass.
Place contaminated broken glass and other sharp objects
(anything likely to puncture an autoclave bag) in a sharps
container designated for autoclaving.
Uncontaminated broken glass does not need to be autoclaved,
but should be disposed of in a specialized broken glass container.