The University of the Virgin Islands
Draft BIOLOGICAL SAFETY PLAN
Biological
hazards are naturally occurring substances such as bacteria, fungi and moulds.
In many cases, these hazards are not as visible or well known as chemical hazards
or physical hazards, such as from fires and electrical shock. As a precautionary measure, laboratory coats,
gloves and, if necessary, masks over the mouth, should be used in order to
avoid personal contact with biological agents.
Disinfections and sterilization procedures should be written out and
applied. Disposal methods must follow to
prevent contamination of the surrounding environment. Practicing good biological safety or “biosafety” procedures is a must for the University of the
Virgin Islands’ faculty, researchers, staff, and students.
SECTIONS:
I.
What is a
Regulated Medical Waste?
II.
Procedure for
Sterilizing or Autoclaving Biohazardous Waste
III.
Prevention of
Acquired Biological Infections
IV.
Biohazardous Waste Disposal
V.
Biological
Hazards Spills
VI.
Personal
Contamination – Removal of Biological Agent From Body
VII. Ethidium
Bromide: Hazards and Precautions and Disposal
VIII. Reporting of Incidents
and Accidents
IX.
Important Response Agencies Addresses and Phone Numbers
I. What is a Regulated Medical
Waste?
Cultures and stocks of infectious agents such as
tissue culture materials including human and primate cell lines, human blood
and blood products, impure animal cell lines, preparations made from living organisms
and their products, including vaccines, cultures, etc. intended for use in
diagnosing, immunizing, or treating human beings or animals, or in research
pertaining thereto.
"Infectious Agents" mean any organisms such as a virus or bacteria that
cause disease or an adverse health impact to humans. Those organisms found in Biosafety Levels 2 through 4 of the CDC's Manual for Biosafety in Microbiological and Biomedical Laboratories
(May 1993) are included.
Culture media
such as agar gel, nutrient broth, discarded media from cultures, and blood
agar.
Clean up materials including absorbents, paper towels from culture/stock mixing.
Biologicals include preparations made from living organisms and
their products, such as serums, vaccines, antigens, antitoxins, etc.
Culture dishes, flasks, or other devices used to transfer, inoculate, and mix culture dishes
such as tissue culture plates, assay plates, test tubes, centrifuge tubes,
cotton swabs, pipettes, pipette tips, stirring apparatus, flasks, vials, beakers,
bottles, jars, spatulas, inoculation loops, wires, corks, stoppers, cell
scrapers, cell lifters, paraffin sealing paper, foil, cotton, filters, mixing
sticks, tubing, etc. that have been in contact with infectious materials.
Bio-Sharps
means discarded unused sharps and sharps used in animal or human research.
Examples include glassware such as Pasteur pipettes, glass, glass culture
dishes, blood vials, glass beakers, glass flasks, glass test tubes, slides,
cover slips, etc. in contact with infectious materials. Plasticware,
including broken rigid plastic items, broken plastic culture dishes/flasks,
plastic pipettes, etc., in contact with infectious materials, are other items
in this category. Syringes to include hypodermic needles and other materials capable
of puncturing such as scalpel blades, suture needles, surgical needles,
lancets, staples, instruments designed for cutting or puncturing: saws,
tweezers, scissors, etc. Glassware and Plasticware
that are heavily contaminated and cannot be fully decontaminated should be
packaged and labeled as Hazardous Solid Medical Waste.
Animal waste
means discarded materials including carcasses, body parts, fluids, blood, or
bedding originating from animals known to be contaminated with infectious
materials.
Medical waste means
discarded materials including carcasses, body parts, fluids, blood, or bedding
originating from humans known to be contaminated with infectious materials.
An evaluation of the biological materials that are
being used is essential to ensure that no RMW is being disposed of in the
regular solid waste. For guidance, "Regulated
Medical Waste Treatment, Storage, Containment, Transport and Disposal"
from the New York State Department of
Environmental Conservation contains the definitions of infectious materials and
procedure for the storage, containment, treatment and disposal of infectious
waste. Anything listed in that guide must be handled and managed as a RMW.
A.
Segregation Requirements for RMW
RMW must be segregated into the following three
categories: sharps, fluids (greater than 20 cc), and other RMW.
Bio-Sharps will be collected for disposal in
leak-proof, rigid, labeled, red puncture resistant containers that are secured
to preclude loss of contents.
Fluid RMW will be housed in autoclaved-able bottles or
glassware, which are impervious to moisture, and have strength sufficient to
resist cracking and bursting under normal conditions of usage and handling.
Other RMW (non-sharps/solid waste) will be collected
in bags, which are impervious to moisture, and have strength sufficient to
resist ripping, tearing, or bursting under normal conditions of usage and
handling. The bags shall be secured to preclude loss of contents and will be
either clear or red in color.
RMW will be collected in each laboratory that
generates it. Laboratories will not store more than one full sharps container
and/or red bag at any one time. When full the sharps containers and/or red and
orange RMW bags will be pick up by Physical Plant for storage prior to pick up
by a licensed company for incineration. The sharps containers will be replaced
with new empty containers at the time of transfer. Non-infectious biological
organisms and associated lab debris should be autoclaved prior to disposal in
the regular trash. Use clear autoclaved-able bags. Infectious biological
organisms and associated lab debris should be autoclaved prior to disposal by a
licensed company for incineration. Use red or orange colored autoclaved-able
bags. Red or orange biohazard bags cannot be placed in the regular solid waste.
Autoclaved fluid of non-infectious biological organisms can be poured down the
sink drain into the public sewer system.
RMW reusable containers must be cleaned with a
disinfectant registered with the United States Environmental Protection Agency
(EPA) or disinfected using a DOH approved non-chemical alternative (e.g., steam
sterilization, radiation, etc.). If any reusable containers used for the
storage or transport of RMW that are unable to be decontaminated, such
containers must be packaged, labeled, treated and disposed of as RMW.
II. Procedure for
Sterilizing or Autoclaving Biohazardous Waste
a. Assure that all waste has been properly separated
by category.
b. Place chemical or biological indicator with load or
autoclave tape on outside of container as means of verifying effectiveness of
sterilization.
c. Use the recommended temperature, pressure and time:
i. Temperature: 1210C to 1230C (2500F to 2540F)
ii.
Pressure: 15 psi or more
iii.
Time: Minimum of 30 minutes or more at the recommended temperature.
d. Make sure that steam is able to contact all areas of
the load:
i. Uncover or open containers or bag to be autoclaved to
enable steam contact with all parts of waste.
ii.
Load must not exceed the effective capacity of the autoclave chamber.
iii.
Make sure steam supply to autoclave is adequate and that there are no air
pockets present in steam line.
iv.
Water can be added to containers if added moisture is needed.
e. Use either SLOW EXHAUST setting for liquids or Use
FAST EXHAUST setting for non-liquids.
f.
Observe autoclave safety
rules to prevent injury such as burns:
i. Use proper protective equipment (lab coat and
heat-resistant gloves) to prevent biohazard exposure and to prevent burns.
ii. Make sure that autoclave door is properly sealed.
iii.
DO NOT open door while the chamber is still pressurized.
iv.
DO NOT put sealed items in the autoclave
g. DO NOT autoclave biohazards containing more than a
TRACE of solvents, volatile or corrosive chemicals (phenol, ether, chloroform,
TCA, etc.).
h. DO NOT autoclave any radioactive material unless sanctioned
to do so by Radiation Safety Officer.
i. Allow autoclave to cool down before opening
door. DO NOT stand in front of its door
when opening it.
j.
Because liquids which are removed too soon tend to boil, avoid the risk of
burns by not overfilling containers and by allowing liquids to cool down
properly before their removal.
k. After sterilization and when bag or box has cooled
sufficiently, close and seal bag or place lid on box. Confirm by inspection of
the heat indicator that sterilization has been successfully.
l. Follow disposal procedures outlined for
decontaminated biological materials and decontaminated non-sharps/solid waste
or sharps.
III. Prevention of
Acquired Biological Infections
Sound
fundamental laboratory techniques can reduce the hazards of infections. Two
important ideas to remember: one, perform procedures to prevent transmission of
infect agent and two, decontaminate any object that
came in contact with the infected agent. The list below outlines procedures
applicable in biological laboratories.
- Sterilize contaminated disposal items, such paper
towels, mask, gloves, pipettes and syringes, after use.
- Decontaminate or autoclave-sterilize contaminated
non-disposal items after use.
- Never pipette liquids with mouth; use a pipetter or bulb.
- Use only pipettes with cotton plugs.
- Do not prepare mixtures of infectious material by
bubbling expiratory air through the liquid with a pipette.
- Use alcohol-moistened cotton around the stopper
and needle when removing a syringe and needle from a ruler stopper vaccine
bottle.
- Use only needle-locking hypodermic syringes.
Avoid using syringes whenever possible.
- Expel excess fluid and bubbles from a syringe
vertically into a piece of cotton moistened with a disinfectant, or into a
small bottle containing cotton.
- Never leave a discarded tray of inflected
material unattended.
- Before centrifuging, inspect tubes for cracks.
Inspect the inside of the centrifuge holders for rough walls caused by
erosion of adhering matter. Carefully remove any bits of glass from the
rubber cushion. A germicidal should be added between the tubes and the
centrifuge holders not only disinfects the surfaces of both of these, but
also provides an excellent cushion against shocks that might break the
tube.
- Use centrifuge tubes with screw caps or
equivalent.
- Avoid decanting centrifuge tubes; if you use it
afterwards wipe of the outer rim with a disinfectant. Avoid filling a tube
to the point that the rim ever becomes wet with culture.
- Periodically, clean out deep-freeze and dry-ice
chests in which cultures are stored to remove broken ampoules or tubes.
Use rubber gloves and respiratory protection during this cleaning.
- Instruct personnel and students to develop the
habit of keeping your hands away from your mouth, nose, eyes, and face.
This may prevent self-inoculation of infectious material.
- Smoking, eating, and drinking are prohibited
inside any biological laboratory.
IV. Biohazardous
Waste Disposal
To assure that minimal harm
to people, other organisms, and the environment will result from the disposal
of biological laboratory waste, the waste disposal program must be implemented
and followed.
A. Liquids
·
Decontaminated biohazardous liquids (such as bacterial cultures in liquid
media, human blood, or animal fluids known to contain pathogens) by treatment
with appropriate chemical disinfectant and steam-autoclaving contaminated
materials.
·
After proper
decontamination, dispose of liquids down the drain or pour into designated
waste disposable containers.
B. Disinfected, Preserved Animal Carcasses and
Fresh Animal Tissues
·
Place animal
carcasses and tissues in a black bag and double the bag if necessary.
·
Spray with
appropriate chemical disinfectant and seal the bag completely and discard in
appointed trash bins.
C. Disposable Contaminated Solids
·
Place solid waste
(such as paper towels, gloves, masks, plastic pipette tips, etc) in clear
autoclave bags seal, and attach a “BIOHAZARD LABEL” and stream autoclaved.
·
After autoclaving,
dispose into appointed trash bins.
D. Sharps and Bio-Sharps (metal sharps or contaminated glass sharps)
·
Place all metal
sharps (such as razor blades, scalpels, needles and syringes) into
designated-labeled, Sharps container; steam autoclave (sterilized) container
before disposal. Place all any
contaminated glass sharps (such as Pasteur pipettes and capillary tubes) in to
the designated-labeled, Bio-sharps container; steam autoclave (sterilized) container
before disposal.
V.
Biological Hazards Spills
If a biological hazard spill
does occur, the following general procedures should be followed:
·
Wear appropriate
personal protective clothing such as gloves, lab coat, and approved respiratory
equipment, if needed, to clean up biological hazard spills.
·
Cover the spill
area with paper towels; pour a 10% bleach solution or disinfectant solution
around the edges of the spill and then into the spill. Allow 10 minutes contact
time.
·
Working from the
outer edges into the center, use paper towels to clean the area.
·
Clean the spill
area with fresh towels soaked in a disinfectant. Be sure to decontaminate any
areas or surfaces that you suspect may have been affected by the spill. Use of
a disinfectant spray on the spill area is appropriated at this point.
·
Place all clean
up materials and gloves into a bag for decontamination, preferably by
autoclaving.
·
If an accident
occurs that may generate aerosols or droplets of an infectious agent, all lab
personnel must leave the area and close the door. Decontaminate clothing and then shower using
disinfectant soap. Allow at least 30 minutes for the droplets to settle and for
the aerosol concentration to decrease before cleanup.
·
Personnel
involved must wash hands and face thoroughly after cleanup.
VI. Personal Contamination
Procedures
for the removal of biological contamination from body.
- Biological Hazard Exposure to Eyes
o
Flush the eyeball
and inner eyelid with cold water for 15 minutes. Forcibly hold the eye open to
wash thoroughly behind the eyelids.
o
Get professional
medical attention promptly.
- Biological Hazard Ingestion and Inhalation
o
Remove all
contaminated clothing.
o
Get professional
medical attention promptly.
- Biological Hazard Exposure on Skin
o
Remove all
contaminated clothing.
o
Immediately flood
the contaminated area with sufficient running water.
o
Wash area with
soap and water and apply disinfectant or 10% bleach solution. Use paper towels to scrub affected area.
o
Remove all
contaminated clothing.
o
Continue to rinse
with cold water for 15 minutes. Again,
wash contaminated areas with the water and disinfectant solution but do not
apply creams or lotions.
o
Place used paper
towels and gloves and contaminated clothing into a biohazard bag to be sterilized
by autoclaving before for disposal.
VII.
Ethidium Bromide: Hazards and
Precautions and Disposal
Ethidium Bromide (EB) is commonly used as a non-radioactive
marker for identifying and visualizing nucleic acid bands in electrophoresis
and in other methods of gel-based nucleic acid separation. EB is a dark red,
crystalline, non-volatile solid, moderately soluble in water, which fluoresces
readily with a reddish-brown color when exposed to ultraviolet light (UV). Its
formula is 2,7,-Diamino-10-ethyl-9-phenyl-phenanthridium
bromide, CAS# 1239-45-8. Although it is an effective tool, its hazardous
properties require special safe handling and disposal procedures.
Hazards
EB is a potent mutagen and
moderately toxic after an acute exposure. EB can be absorbed through skin, so
it is important to avoid any direct contact with the chemical. EB is also an
irritant to the skin, eyes, mouth, and upper respiratory tract. It should be
stored away from strong oxidizing agents in a cool, dry place, and the
container must be kept undamaged and tightly closed.
Safety Precautions
People using EB should follow
several safety procedures. The laboratory's Chemical Hygiene Plan should
reference this Fact Sheet, which outlines safe handling of EB and proper
cleanup procedures. EB users should receive documented safety training on its
hazards. EB must appear on the laboratory's chemical inventory, with accurate
estimates of on-hand and yearly use quantities.
- Pure EB should only be handled in a fume hood,
with the user wearing protective equipment that includes a lab coat,
closed-toe shoes, chemically resistant gloves, and chemical safety goggles
(not just safety glasses).
- Nitrile and latex gloves are effective barriers to
short-term exposure to EB.
- Chemical safety goggles must be worm when
handling EB.
- EB users should wash their hands after removing
their gloves, even if they are certain the gloves weren't punctured. An
emergency eyewash and shower should be accessible nearby.
- Like all other toxics, EB should be used in a
specially designated area where no eating or drinking is allowed.
- When using ultraviolet light to visualize EB, the
user must wear UV-blocking eyewear or work in a UV cabinet with shielding
glass in place.
Emergency Exposure Procedures
If EB contacts the eyes,
immediately flush them with copious amounts of cold water for at least 15
minutes. (If it is available, emergency eyewash is the best and safest way to
do this.) For skin contact, immediately wash the affected area with soap and
copious amounts of cold or cool water. If a person inhales EB dust, move him to
an area where he can breathe fresh air. After any exposure to EB (via skin,
inhalation, or eye contact), the affected person should immediately seek a
medical evaluation.
Spill Procedures
Notify all others in the room that the spill has
occurred. Evacuate the room or immediate area and call research PI for
assistance with the cleanup Post the room with signage warning others of the
spill, and prevent unnecessary entry into the . Always wear full protective
clothing, as described above, during any cleanup procedure. You should only
clean up spills if you are aware of the hazards and the decontamination
procedure and have the proper safety and cleanup. A portable UV light is also helpful for
checking to see if surfaces or other items have been contaminated with EB; the
user must wear UV-blocking eyewear and gloves when operating this light.
Disposal
Unwanted solid EB, gels, and
all working solutions must be collected and place into the Ethitidum
Bromide Waste Container. The container with words that clearly identify the
contents as Hazardous Waste and affix completed Hazardous Identification Waste
Tag. Ethitidum
bromide waste must be disposed via a certified chemical waste disposal company.
VIII. Reporting of Incidents and Accidents
Reporting of accidents and
occupational illnesses involving death, critical injury, lost time health care
(by a medical practitioner) or damage to University’s property is required for
employees, for students, and for visitors under both the Occupational Health
and Safety Act and the Workplace Safety and Insurance Act. Reporting of accidents is done centrally
through the Office of Environmental Health and Safety. Employees should understand that the purpose
of reporting and documenting accidents is not to affix blame, but instead to
determine the cause of the accident so that similar incidents may be prevented
in the future. Incidents should be reported on the Incident Report Form within 48
hours of occurrence. Accidents should be reported on the either
an Injury Accident Report Form and Non-Injury Accident Report Form
within 48 hours of occurrence. Period
reviews of these reports will often reveal problem areas that need special
attention
·
Reportable incidents and accidents are those which:
o
result in personal injury or lost time from work (excluding
those requiring minor first aid).
o
result in property damage.
o
occur to a university employee during the course of his/her
work either on or off university premises.
o
occur to a student during the course of his/her classroom,
laboratory or field work.
o
occur to a student during the course of a work placement
(either paid or unpaid) which forms part of their university curriculum.
o
occur to any person on university premises.
IX. Important Response
Agencies Addresses and Phone Numbers
Fire Emergency 911
Police Emergency 911
University of the
University of the
Government of the
Virgin Islands, Department of Planning & Natural Resources, Environmental
Protection,
Government of the
Virgin Islands, Department of Planning & Natural Resources, Environmental
Protection,
Environmental
Protection Agency, Federal Building Room 142,
Centers for Disease Control and
Public Inquiries: 1-800-311-3435 or 404-639-3534
National Response
Center, c/o United States Coast Guard (G-OPF), 2100 2nd Street, Southwest -
Room 2611 Washington, DC 20593-0001
Chemical/Biological Hotline: 1-800-424-8802
Occupational Safety &
Health Administration, Region 2, Regional Office for U.S. Virgin Islands, 201 Varick Street, Room 670, New York, New York 10014 212-337-2378
Written by Ann Marie Dublin and Eugenia Somerall-Sello
Reviewed by Paul L. Kahn, On-Scene
Coordinator, Environmental Protection Agency
Web Page Designed by Eugenia Somersall-Sello
Last Updated: April 29, 2007