RULES FOR WORKING WITH MICROORGANISMS
Precautionary Proceedures
- Do not eat, drink, smoke, apply cosmetics or lip balm, or store
food in the laboratory.
- Bring any necessary equipment, laboratory instructions and your
lab notebook to the work bench. A flow chart and/or notes may help
you carry out the lab work with a minimum of page shuffling.
- Wear a protective laboratory coat or apron when you are working
with cultures, and avoid wearing long, full sleeves if possible.
- Tie long hair back or put it up. If hair hangs loose, it becomes
a contamination hazard and also may catch on fire in the bunsen burner
flame.
- Carry and store cultures of microorganisms in racks or baskets.
Do not leave cultures on the table or in unmarked areas when the laboratory
session is completed.
- Place cultures to be discarded in racks or trays designated for
contaminated material; these racks should be clearly labeled. All
such materials should be autoclaved before further handling, discarding,
or washing.
- Decontaminate work surface after spills and at the beginning and
end of each laboratory period with the disinfectant provided in the
laboratory.
- Cover small spills with paper towels and soak the towels well with
disinfectant. Let the towels stand for one-half an hour. Place these
materials in a container with disinfectant or in a plastic bag which
can be sealed. Autoclave container before discarding.
- Mix liquid cultures gently to avoid foaming and splashing which
may produce an aerosol. Do not mix cultures by bubbling expiratory
air through the liquidwith a pipette.
- Never pipette cultures by mouth; mechanical pipetting devices are
to be used. Place contaminated pipettes in a pan which contains an
appropriate disinfectant.
- Wash hands carefully with soap after any possible contamination
and before leaving laboratory. Dry hands thoroughly after washing.
- Open cuts should be covered with bandages; if they are on the hands,
wear disposable gloves.
- Individuals who have special health problems such as diabetes or
suppressed immunity from disease or therapy should be encouraged to
discuss them privately with the instructor.
- Develop the habit of keeping your hands away for your mouth, nose,
eyes, and face to prevent self-inoculation.
- Before centrifuging, inspect tubes for cracks, Whenever possible,
use autoclavable plastic centrifuge tubes with screw caps. Avoid filling
the tube to the point that the rim becomes wet with the culture.
- Avoid the use of hypodermic syringes. Use a pipette whenever possible.
If a syringe is needed, use a needle-locking hypodermic syringe.
- Before and after injecting an animal, swab the site of injection
with disinfectant.
- Shake broth cultures in a manner that avoids wetting the plug or
cap.
- Periodically clean out deep freeze and dry-ice chests in which cultures
are stored to remove broken ampules and tubes. Use rubber gloves and
respiratory protection during the cleaning.
- Handling of materials infectious to man and other animals in the
laboratory require theAugust 9, 2006 federal, state and local government.
Emergency Procedure For Working With Microorganisms:
- Spills
In the case of spill, flood area with disinfectant provided in
the laboratory. Dispose of paper towels with contaminated waste. Wipe
counter dry with paper towels.
For spills on carpeting, after flooding with disinfectant, clean up
as directed by the instructor.
- Ingestion
Do not swallow.
Empty contents of mouth into sink.
Rinse mouth with copious amount of water.
Report all ingestion to instructor.
- Exposure
Eyes: Immediately WALK to eye wash station. Turn on water.
Hold eyelids open and wash for at least two minutes. Inform instructor.
Cuts: Wash cut with water. Follow with disinfectant. Place
a pad or clean material over the cut. Apply pressure directly over
cut. Do not put mouth over the cut. Do not breathe on the cut.
Needle injection: Report immediately to the instructor. The
instructor will call infirmary and the infirmary will then determine
the appropriate course of action.
Biosafety Level 2
Biosafety Level 2 is suitable for experiments involving agents
of moderate potential hazard to personnel and the environment. It
differs in that laboratory personnel have specific training in handling
pathogenic agents and are supervised by competent scientists. Access
to the laboratory is limited when experiments are being conducted,
and procedures involving large volumes or high concentrations of agents
or in which aerosols are likely to be created are conducted in biological
safety cabinets or other physical containment equipment.
Special Practices
Contaminated materials that are to be decontaminated at a site away
from the laboratory are placed in a durable leak-proof container which
is closed before being removed from the laboratory.
- Access to the laboratory is limited by the laboratory supervisor
when experiments are being conducted. In general, persons who
are at increased risk of acquiring infection or for whom infection
may be unusually hazardous are not allowed in the laboratory or
animal rooms. Persons at increased risk may include children,
pregnant women, and individuals who are immunodeficient or immunosuppressed.
The supervisor has the final responsibility for assessing each
individual circumstance and determining who may enter or work
in the laboratory.
- The laboratory supervisor will assure that only persons who
have been advised of the potential hazard and meet any specific
entry requirements (e.g., immunization) may enter the laboratory
or animal rooms.
- When infectious materials or infected animals are present in
the laboratory or animal rooms, a hazard warning sign, incorporating
the universal biohazard symbol, is posted on all laboratory and
animal room access doors and on such other items (i.e., equipment,
containers, materials) as appropriate to indicate the presence
of viable infectious agents. The hazard warning sign should identify
the agent, list the name of the laboratory supervisor or other
responsible person(s), and indicate any special requirements for
entering the area (immunization, respirators, etc.).
Biohazard Warning Symbol:
An insect and rodent control program is in effect.
- Laboratory gowns, coats, or uniforms must be worn in the laboratory
but must not be worn to a lunchroom or outside the building in which
the laboratory is located.
- Animals not involved in the experiment being performed are not
permitted in the laboratory.
- All wastes from laboratories and animal rooms must be appropriately
decontaminated before being disposed.
- The use of hypodermic needles and syringes is restricted to
lavage, parenteral injection and aspiration of fluids from laboratory
animals and diaphragm vaccine bottles. Hypodermic needles and
syringes are not used as a substitute for automatic pipetting
devices in the manipulation of infectious fluids. Serial dilutions
of infectious agents should not be done in diaphragm bottles with
needles and syringes because of the hazards of autoinoculation
and of aerosol exposure. Cannulas should be used instead of sharp
needles whenever possible.
- If activities of lesser biohazard potential are conducted in
the laboratory concurrently with activities requiring Biosafety
Level 2, all activities will be conducted at Biosafety Level 2.
- Gloves should be worn for all procedures requiring the handling
of infectious materials or infected animals. Holding small laboratory
mammals with forceps when they are receiving injections or otherwise
being handled provides an additional level of protection for personnel.
- Serological procedures with inactivated antigens shown to be
free of residual infectivity can be performed on the open bench.
- All spills, accidents, and overt or potential exposures to infectious
materials must be immediately reported to the laboratory supervisor.
A written record must be prepared and maintained. Appropriate
medical evaluation, surveillance, and treatment
- Baseline serum samples should be collected from and stored
for all laboratory and other at-risk personnel. Additional serum
specimens may be collected periodically depending on the agents
handled or the functions of the facility.
- A safety or operations manual which identifies known and potential
hazards and which specifies practices and procedures to minimize
or eliminate such risks should be prepared or adopted. Personnel
should be advised of special hazards and are required to read
and follow standard practice and procedures.
Containment Equipment
Biological safety cabinets (Class I, II, or III) or other appropriate
personnel protective or physical containment devices are used whenever:
- Procedures with a high potential for creating aerosols are
conducted. These may include centrifugation, grinding, blending,
vigorous shaking or mixing, sonic disruption, opening containers
of infectious materials whose internal pressures may be different
from ambient, intranasal inoculation of animals, and harvesting
infected tissues from animals or eggs.
- High concentrations or large volumes of infectious agents are
used. Such materials may be centrifuged in the open laboratory
if sealed heads or centrifuge safety cups are used and if they
are opened only in a biological safety cabinet.
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