Contents
Forword
This manual is designed to provide general operating and
emergency procedures for working safely in the biohazard facility (BHF) located in the
Chanin Building. This manual also defines the duties and responsibilities of all
individuals using or operating the BHF.
The BHF, located on the sixth floor of the Chanin
Building, is designed to meet federal requirements for a Biosafety Level 3 (BSL-3)
containment laboratory. Containment of biohazards is achieved through the use of primary
control equipment, facility design, and laboratory procedures. Not all laboratories or
animal rooms in the BHF are classified as BSL-3. Laboratories or animal rooms considered
BSL-3 will be clearly marked at their entrance.
Biohazard containment in this facility will function
properly if operational procedures are followed. The operational procedures described in
this manual have been designed to provide a high degree of protection to our workers and
to the community from potentially infectious materials that may be used in this facility.
We would like to emphasize that even the best containment facility and equipment is
rendered ineffectual with improper laboratory procedures. Persons working in the biohazard
facility must be trained and proficient in microbial practices and techniques before
handling infectious or potentially infectious materials. The Principal Investigator is
directly responsible for ensuring that each employee has received appropriate training and
experience before beginning work in the BHF.
The operating procedures detailed in this manual shall
apply to all staff, support personnel, and to any authorized visitors to the facility. It
is essential that all personnel entering any laboratory in the BHF read and comply with
this manual.
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I. Roles and Responsibilities
INSTITUTIONAL BIOSAFETY COMMITTEE
The Institutional Biosafety Committee (IBC) approves or
rejects all proposals and research conducted within the biohazard facility (BHF) and also
designates the biosafety level required. It is also responsible for: reviewing activities
which raise health and safety issues, reviewing the activities of the Environmental Health
and Safety Department as it pertains to infectious agents, assess containment levels,
establishes a medical surveillance plan for all appropriate personnel and reviews any
changes, challenges or grievances concerning research within the facility.
BIOSAFETY OFFICER
The Biosafety Officer (BSO) is the administrator of the
BHF. The BSO has the authority to: determine if an employee is unable to work within the
facility, deactivate any malfunctioning containment equipment, and insure compliance with
governmental health and safety regulations. The BSO is responsible for coordinating
meetings of the IBC, provide technical guidance and training materials to personnel
regarding laboratory safety, revise day-to-day procedures as experience dictates, insure
that workers follow procedures and practices, advise Principal Investigators as to the
proper functioning of their workers, initiate and supervise any needed emergency response,
investigate and report to the IBC and Principal Investigator, any significant violations
within the facility, accompany authorized visitors or maintenance workers around and into
the BHF.
PRINCIPAL INVESTIGATOR
The Principal Investigator (PI) is immediately responsible
to insure that the purpose of this manual and all other applicable guidelines are
fulfilled. He should verify that all staff members conducting research within the BHF are
properly trained, have read the BHF manual, and follow the specific protocols and policies
related to the containment laboratories. The PI and his specific research projects must
inform the laboratory staff of any potential hazards associated with their work including,
biological, chemical, and radioactive hazards. The PI is responsible for investigating and
reporting to the BSO, in writing, any accidents or incidents involving his staff in the
BHF. The PI must also notify the BSO and the IBC of new employees who will be working in
the BHF. All new employees must meet with the BSO before they are assigned to work in the
BHF.
INDIVIDUAL LABORATORY WORKER
All personnel assigned to the BHF should read and comply
with the procedures of this manual and meet with the BSO before starting work. In addition
they should be clearly instructed by their PI as to the procedures they must follow while
performing research in the BHF. The laboratory worker is responsible for properly labeling
all biological, chemical and radioactive materials within the facility. Any unsafe act or
malfunctioning equipment should be brought to the immediate attention of the BSO and the
PI. Employees should report to their PI and the BSO any instances which constitutes an
exposure to biological, chemical or radioactive materials. Individuals who are pregnant or
immunocompromised should seek medical advise before working in a BHF laboratory.
AUTHORIZED USER
An authorized user of the BHF is an individual who has
sufficient training and experience to work safely in this facility. The authorized user is
selected by the Principal Investigator assigned to the BHF and is approved or authorized
to work in the area by the BSO and the IBC. This authorization can be rescinded.
Periodically, the Principal Investigator will provide a list of all authorized users
permitted in the BHF.
INSTITUTE FOR ANIMAL STUDIES
The Institute for Animal Studies is responsible for all
research using animals. The Institute for Animal Studies reviews all research projects
using animals, inspects animal rooms, maintains animals, posts hazard warnings on
entrances to rooms, provides training to animal handlers and assists with the proper
disposal of animal waste.
INSTITUTION
The Institution assumes the responsibility for insuring
compliance with all guidelines within this manual and other applicable guidelines
including:
- CDC/NIH: Biosafety in Microbiological and Biomedical
Laboratories, 3rd edition, 1993.
- NIH - Working Safely with HIV in the Research Laboratory
Biosafety Level, 2/3, 1988.
- CDC Guidelines for protection against Viral Hepatitis and
Hepatitis B prevention.
- NCI Biological Safety Manual for Research Involving
Oncogenic Viruses.
- CDC/NIH: Primary Containment for Biohazards: Selection,
Installation and Use of Biological Safety Cabinets, 1st edition, 1995.
- NIH Guidelines on recombinant DNA Molecules, October 1997.
- OSHA Rule governing Occupational Exposure to Bloodborne
Pathogens, 1990.
- CDC guidelines to prevent T.B. exposure.
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II. Physical Containment
Feature and Zone Classification:
The term "containment" is used in describing
safe methods for managing infectious agents in the laboratory environment where they are
being handled or maintained. The purpose of
containment is to reduce or eliminate exposure potential
or risk of laboratory personnel and others, and to prevent escape of potentially
infectious agents to the outside environment. Primary containment, the protection of
personnel and the immediate laboratory environment from exposure to infectious agents, is
provided by good microbiological technique and the use of appropriate safety equipment.
Secondary containment, the protection of the environment external to the laboratory from
exposure to infectious materials, is provided by the combination of facility design and
operational practices.
The biological safety cabinets (BSC) are among the most
effective, as well as the most commonly used, primary containment devices in laboratories
working with infectious agents. These cabinets when used in conjunction with good
microbiological techniques, provide an effective containment system for safe manipulation
of moderate and some high-risk microorganisms.
The two types of laminar flow biological safety cabinets
utilized in the BHF are the Class II type B and Class III cabinets. The Class II cabinet
is the most common and can be found in each laboratory. This BSC is designed for work with
low to moderate risk agents, up to BSL3. The design features of this hood include an air
barrier along the work opening to prevent the escape of biological agents into the
laboratory, High Efficiency Particulate Air (HEPA)-filtered supply air across the work
surface, and HEPA-filtered exhaust air.
The Class III cabinet, located in room 622 is designed for
work with hazardous agents assigned to BSL3 or 4. This cabinet provides maximum protection
to the worker and environment. The Class III BSC is a totally enclosed, ventilated cabinet
of gas-tight construction with a non-opening view window. This cabinet operates under
negative pressure and both supply and exhaust air is HEPA-filtered with the exhaust
passing through a double HEPA filtration before being discharged. Manipulation of
materials with the cabinet is through heavy-duty rubber gloves attached in a gas tight
manner to ports in the cabinet. This system prevents direct contact with hazardous agents
which are introduced into the cabinet via a double door pass-through box.
Secondary barriers are provided by special laboratory
design features. A bioseal on the perimeter of the facility forms an integral shell which
can be decontaminated, when needed. All penetrations into the facility are sealed on both
the clean and contaminated side of the room. The air-handling system in the facility is
balanced such that the facility is negative to the rest of the building, the laboratories
and animal rooms are negative with respect to the corridor, and the biological safety
cabinets are negative with respect to the laboratories. Thus, the direction of the air
flow is always toward the area of increasing hazard. Exhaust air from the facility is
HEPA-filtered before being released to the environment.
Contaminated waste is removed from the facility via the
pass-through autoclave. Animal carcasses and decontaminated materials are then transported
to a pathological incinerator where materials are completely destroyed. All exhausts,
including vents, and vacuum lines are all equipped with in-line HEPA filters.
Personnel access and egress for the facility is through
the access corridor located at the north and south ends of the facility. This access
corridor may also be used for the introduction of materials and equipment through the
secondary barrier.
The facility is divided into two zones, potentially
contaminated zone and non-contaminated zone. Potentially contaminated zone is the primary
containment zone, which includes the class II and class III laminar flow biological safety
cabinets constitute. All work requiring BSL-2 or 3 containment must be conducted within
these cabinets.
The open laboratory consists of the space exterior to the
BSC which is also considered potentially contaminated and constitutes the secondary
containment zone. All work conducted in this space shall be in accordance with BSL-2 and 3
requirements. Another potentially contaminated zone also includes: the interior of the
facility beyond access corridor, the ventilation system up to and including the HEPA
filters and the sewage system.
The BHF interlocks at the North and South Corridor serve
as a transition zone between the potentially contaminated and non-contaminated zones. The
non-contaminated zone is associated with the area "external" to the interlocks.
Entry into the interlocks is restricted to authorized personnel.
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III.
Facility Assignment Procedures:
Any investigator desiring use of the facility shall submit
a written research proposal to the IBC and the BSO. This proposal must have the signature
of the individuals departmental chairman. Space in the BHF is allocated on a
temporary basis and only to those persons who have completed authorized user
certification. This certification includes, but is not limited to, an orientation to the
BHF and a review of procedures while working in the facility. The IBC will approve use of
biohazard materials within the BHF. Biohazard material may not be used for any purpose, or
in any other location, other than that originally approved by the IBC. Should a new
project be initiated, a new written proposal should be submitted to the IBC and BSO.
Personnel changes are to be reported to the BSO as they occur. Once a year the BSO will
generate a list of certified personnel and request that the PI verify that it is current.
Upon completion of a project, the investigator is required
to decontaminate and remove all materials and equipment in accordance with established
procedures. The BSO will ensure that all materials and equipment have been decontaminated
prior to removal from the BHF laboratories.
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IV. Medical
Considerations:
A. PRE-PLACEMENT
All persons working in the Biohazard Facility have the
option of participating in a pre-placement/annual serum collection program. Please discuss
this option with the Principal Investigator or the Biosafety Officer.
B. MEDICAL RESTRICTIONS
Pregnant women, persons on antacids, steroid therapy, or
immunosuppressive drugs shall not work in the facility prior to a thorough evaluation of
the risks involved. The decision to allow these persons to work in the facility is to be
made by an appropriate physician and the responsible PI with notification to the BSO.
Persons with a fresh or healing laceration or skin lesions
should not work with infectious material unless the injury is completely protected.
Personnel with injuries of this type must notify their PI and the BSO prior to working in
the facility.
C. REPORTING
Emergency telephone numbers must be posted near entrances
and at the telephones in every laboratory. Telephone numbers must include the PI, lab
contact person, Environmental Health and Safety office and an emergency number.
Any illnesses or symptoms known to be associated with
organisms, chemicals being used, or any change in medical condition should be immediately
reported to the PI and BSO.
An incident report is to be completed by the investigator
and forwarded to the Biosafety Officer as soon as possible after the occurrence of the
following:
- Physical injury - e.g., cut, burn, broken bone, slip/fall,
other.
- Hazardous materials exposure - contamination through
parenteral exposure to a biohazardous material, exposure to a hazardous aerosol, ingestion
of a contaminant, or exposure to a carcinogenic compound by inhalation, ingestion, or skin
absorption.
- Any spill involving a hazardous, chemical, biological, or
radioactive material.
D. MEDICAL PROTECTION
Hepatitis-B vaccine is offered through Occupational Health
to all "at-risk" personnel and is provided to the worker at no cost. Your
Principal Investigator will determine if you are at risk of exposure to Hepatitis-B.
The BSO shall provide to all authorized users of the BHF,
current information on the availability of any medical protection (i.e., vaccines)
appropriate for the infectious agents used in the facility. This protection shall be
offered to all "at-risk" personnel.
E. MEDICAL FOLLOW UP
In the event of an accidental parenteral exposure to HIV
and/or HBV-related materials, perform immediate first aid and promptly report to
Occupational Health. The Hepatitis-B vaccine will be offered and/or serology tested. With
the individuals written consent a baseline HIV serology will be drawn, post exposure
chemoprophyaxius will be administered and HIV counseling will be provided. The
Occupational Health Physician will determine and establish the appropriate medical
interventions necessary to treat the exposure.
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V.General
Laboratory Safety:
A. EQUIPMENT
Equipment to be utilized in the facility must meet
building electrical safety standards, which refer to local and national codes. It is
critical that equipment to be selected which:
- Does not contribute to spread of biohazards.
- Does not present a hazard to personnel or facility during
operation.
It is essential that all equipment be properly maintained.
If equipment malfunctions it should not be used and the BSO should be informed. Any
equipment exiting the BHF must be decontaminated under the supervision of the BSO.
Responsibility for cost, decontamination, or repair of equipment in individual
laboratories falls to the Principal Investigators.
B. PHYSICAL BARRIER SYSTEMS
The facility is equipped with Class II and III Biological
Safety Cabinets which are designed to provide protection for personnel as well as
materials within the cabinets.
The facility and its installed equipment are designed to
provide physical barriers for personnel against exposure to biological hazards. The
ventilation system is designed and adjusted to provide directional flow toward the area of
increasing hazard. The general ventilation systems are HEPA-filtered to entrap infectious
agents and to prevent dispersion outside of the BHF.
C. BIOLOGICAL AGENTS
All infectious and/or biologically hazardous agents being
used in the BHF facility must have approval of the IBC and be registered with the BSO. A
hazard warning sign incorporating the universal biohazard symbol should be posted on all
the access doors into the laboratories or animal rooms. The hazard warning sign should
identify the infectious agent, list the name and telephone number of the PI or other
contact person, and indicate the special require-ments for entering the laboratory. In
addition, each laboratory or animal room will be given either a BSL-2 or BSL-3 designation
on the entrance door. Transportation of infectious materials within the facility must be
in a secured leak-proof, unbreakable container with an appropriate label. An absorbent
material should be added to the container in case of breakage.
Procedures for handling and disposal of these agents are
outlined below.
D. CHEMICALS
The handling of chemicals in the facility requires the
exercise of proper controls in terms of the class of chemical and any associated hazard.
As a general practice, the quantities of chemicals introduced into the facility should be
kept to the minimum required for immediate operations. Excess storage of hazardous
chemicals in the BHF should be avoided.
The disposal of acids, alkalis, and organic solvents into
the sewer system is prohibited. Chemical waste should be containerized, surface
decontaminated and appropriately labeled. The BSO should be contacted for proper disposal
procedures.
Waste materials, (i.e., gloves, absorbent pads, culture
flasks, etc.) shall be placed in autoclave bags and autoclaved. Liquid, infectious wastes
shall be placed in a sealed container with a disinfectant, appropriately labeled, and
surface decontaminated. Notify the BSO for proper disposal.
E. RADIOISOTOPES
The introduction and the use of radioisotopes in the
facility must be in accordance with regulations established by the AECOM Radiation Safety
Officer (RSO). The Principal Investigator must be authorized to use the radioisotope with
approved handling, safety, and waste disposal procedures. Every attempt should be made to
minimize the generation of mixed hazardous waste (infectious and radioactive) unless there
is a clear, easy, and safe way to inactivate one of the hazardous components. Under no
circumstance should mixed radioactive and infectious waste be transferred to the RSO
without prior neutralization of the biological agent.
See Emergency Procedures involving Radioactive Material
F. ACCIDENTS AND INJURIES
The specific procedures regarding management and reporting
of accidents or injuries shall be in accordance with AECOM rules and regulations. The PI
should ensure that all personnel associated with the program are cognizant of these
procedures. Any questions should be directed to the BSO. These procedures can be found
below.
See
Emergency Procedures Below
G. FIRE
The specific procedures regarding management and reporting
of fire shall be in accordance with AECOM rules and regulations. The PI shall ensure that
all personnel assigned to the facility are cognizant of these procedures.
See
Emergency Procedures Below
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VI. Safety Procedures:
The BSL2 and 3 facility is specifically designed to
conduct work involving biological material of potential hazard. All personnel must be
advised of the potential hazards and instructed in the operational procedures of the
facility and specific laboratory. Only persons authorized on the basis of program or
support needs shall enter the facility. All personnel admitted into the facility shall
read, understand, and follow the procedures detailed in the manual. Any questions
regarding safety procedures in the BHF must be addressed to the Principal Investigator,
Institute for Animal Studies or Biosafety Officer.
PERSONNEL PRACTICES
At the start, build safety into your procedures to
minimize the likelihood of an adverse outcome.
Working in a relaxed, un-anxious, and non-hurried manner
may help you to anticipate and avoid problems.
- A Tyvek® suit, disposable lab coat or
surgeons gown shall be worn at all times in the BHF laboratories.The BSO mayrequire a disposable Tyvek® suit and other protective equipment to be worn in
various other locations within the BHF.
- Reusable lab clothing must not enter the BSL-2 and 3
laboratories. They must be stored in lockers (if available) or hung on hooks located
within the interlocks or laboratory anti-chamber.
- Disposable gloves, provided by the PI, will be worn in the
facility when handling biohazardous agents. The gloves should be taped to the cuff of the
protective clothing to prevent any skin exposure. Powderless latex gloves are recommended
for use when handling infectious material.
- Double gloving is required before penetrating the
biosafety cabinet and removed on the way out of the cabinet.
- Special care shall be taken to avoid skin contamination
with infectious materials and this could be a major route of personal exposure.
- Hands should be washed frequently during the day using the
foot-operated faucet and germicidal soap provided in each laboratory. Ordinary soap and
water is also effective in cleansing the hands.
It is mandatory to wash hands:
- After handling infectious materials.
- After a spill and appropriate clean up.
- When removing protective gloves.
- When exiting the laboratory.
- When exiting the facility.
Respiratory protection may be recommended when there is
the possibility of hazardous aerosol generating procedures or as deemed necessary by the
BSO. If you must wear a respirator, then you must be part of the Respiratory Protection
Program which includes a medical evaluation and a fit test.
- Head coverings may be required, particularly, with long
hair. Long hair should be covered or pinned up.
- Contact lenses do not provide eye protection. It is
strongly recommended that contact lenses not be worn while conducting work in the
laboratory. Safety goggles with side shield, goggles, or a plastic face shield should be
worn for eye protection.
- Smoking, food, beverages, and cosmetics are not permitted
in the BHF.
- Mouth pipetting, under any circumstances, is not
permitted.
- Disposable cleaning tissue should be used rather than
personal handkerchiefs.
- Personnel must notify the PI and the BSO of any event or
incident which may compromise the safety of personnel or work.
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VII.
Emergency Telephone Numbers
EMERGENCY (All Hours) Dial X 4111
Includes:
- Fire
- Biohazard Spill
- Chemical Spill
- Radioisotope Spill
- Medical Emergency
State the nature of the emergency and the location. If
possible, remain nearby to direct the emergency response team.
Biosafety Officer X3560
- Health and Safety Office X4150
- Radiation Safety Office X2243
- Engineering X3000
- Security X2019
ALWAYS NOTIFY THE SAFETY OFFICE IN AN
EMERGENCY OR ACCIDENT!!!
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VIII.
Emergency Procedures
INJURIES AND ILLNESSES
Serious Injury or Sudden Illness:
- Dial the emergency number 4111, when special first aid,
resuscitation, transport, or rescue service is required. Clearly describe the situation
and your location.
- Place all contaminated materials in either a biological
safety cabinet or appropriate containment so that medical help can enter the facility.
- Notify the PI and BSO.
Minor Injuries:
- Report all incidents to the PI and BSO.
- A first aid kit is located on the wall in the access
corridor near the eyewash station. The use of the first aid kit does not preclude a visit
to Occupational Health.
- Obtain an "Accident/Incident Report Form" from
the PI, and report all injuries to Occupational Health for treatment and the BSO
Emergencies include, but are not limited to, a
biohazardous or hazardous chemical spill, fire, BSC malfunction, or a total power failure.
The primary objective in an emergency is preservation of personal safety and health.
Protecting the facility and the experiment are secondary to personal safety. If there is a
hazardous spill in your work area and you are not wearing a respirator, hold your breath
and evacuate the room.
BASIC PRINCIPLES
Immediate personal safety overrides maintenance of
containment. Evacuation takes priority. Get people out of the emergency area. If
possible biohazardous materials should be covered and contained. All equipment should be
turned off. The BSO must be informed as soon as possible and will take charge of
re-entry, clean-up, and other corrective measures.
The Investigator or BSO is responsible for deciding
whether to override containment procedures in case of serious injury or sudden illness.
It is essential that the authorized users of the BHF
familiarize themselves with the procedures detailed here. Questions about these procedures
should be directed to the PI and the BSO. BHF personnel should be aware of all exits,
interlock override switches, fire extinguishers, fire alarms, eyewash stations, safety
showers, spill and first aid kits. KNOW WHAT TO DO BEFORE AN EMERGENCY OCCURS.
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IX. Biohazard Spill Outside a Biological Safety Cabinet(BSC)
If Biohazardous material is spilled in the open
laboratory, one must avoid inhaling any airborne infectious material and getting the
infectious agent onto your body and clothing. A "spill kit" is available in the
facility supply closet, C-673. You should always have available, a freshly prepared
solution of a disinfectant in the event of a sudden spill. The BSO should be notified once
the contaminated laboratory has been evacuated. Others in the area are to be warned
against entry.
Immediate Spill Control:
- Dont breathe, evacuate all personnel and close the
door.
- Remove contaminated clothing carefully, folding the
contaminated area inward. Place clothing in a bag or directly into the autoclave.
Thoroughly wash hands and face and any exposed area of the body. Shower, if necessary.
- Notify the BSO and PI.
- POST SIGNS WARNING OTHERS NOT TO ENTER CONTAMINATED AREA.
NO ONE SHOULD ENTER THE ROOM FOR AT LEAST ONE HOUR. (This allows aerosols to be carried
away and heavier particles to settle.) Time should be taken to formulate a plan to
decontaminate. Once all personnel have been removed from the area, there is no need to
rush into the contaminated area.
- Assist the BSO as necessary. Decontamination will involve
treatment of gross contamination by local application of disinfectant and possible gaseous
decontamination of the entire working space.
- Gaseous
decontamination listed below
Decontamination of a Spill:
- Re-entry into the facility must be delayed for a period of
at least one hour to allow reduction of the potential aerosol generated by the spill.
- Dress in protective clothing, including a Tyvek® suit and
double gloves. Respiratory protection is strongly recommended and care should be taken
during decontamination not to disperse droplets.
- Place paper towels along the outside of the spill, working
from the edges in. Pour the germicidal solution (10% solution of sodium hypochlorite
(household bleach) or Vesphene II se®) around the spill and allow to flow into
the spill. To prevent aerosols, avoid pouring the germicidal solution directly onto the
spill. Try covering the spill with an absorbent pad and apply the decontaminant to the
absorbent pad.
- Allow to stand for 30 minutes, this will provide enough
contact time for adequate disinfection.
- Carefully remove the soaked pads, placing them into an
autoclave bag. Working toward the center of the spill, use paper towels to wipe up the
spill. Discard paper towels as they are used into an autoclave bag.
- Using paper towels soaked in disinfectant, wipe beyond the
area of visible or suspected splashing, including the floor and vertical surfaces. Discard
paper towels in the autoclave bag.
- Decontamination is complete when the whole area of
suspected liquid contamination has been washed with a disinfectant and all excess
decontaminate has been mopped up.
- Place all contaminated materials including gloves,
shoecovers, and other protective clothing into an autoclavable bag. Sterilize and dispose
of this waste in the red bag system as medical waste.
When the above procedure is followed, the spill area is
considered to be decontaminated. The BSO and/or the PI will determine whether the entire
laboratory area requires gaseous decontamination.
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X. Biohazard Spill Inside a Biological Safety Cabinate(BSC)
A spill that is confined to the interior of the BSC should
present minimal or no risk to to personnel in the area. However, chemical disinfectant
procedures should be initiated at once while the cabinet ventilation system continues to
operate to prevent escape of contaminant from the cabinet.
- Spray or wipe, wall, work surfaces and equipment with a
disinfectant. A 10% solution of sodium hypochlorite (household bleach) or Vesphene II se®
is recommended. The operator must be properly gloved and gowned during this procedure.
Household bleach can penetrate latex gloves and can be corrosive to metal so consider
having an alternative available.
- Flood the work surface of the BSC with sufficient
disinfectant solution to ensure that the drain pans and catch basins below the work
surface contain the disinfectant. Allow the disinfectant to work for 30 minutes before it
is cleaned up.
- Make sure to wipe all surfaces including the front intake
grill. Drain the disinfectant into a container.
- Repeat above process with distilled water or mild soap and
water.
- The disinfectant, gloves, wiping cloth and sponges should
be discarded into an autoclave bag, the material should be autoclaved and discarded in the
red bag system as medical waste.
- This process will not disinfect the filters, blower, air
ducts, or other interior parts of the cabinet. The BSO should be consulted to determine if
gaseous decontamination of these items is necessary.
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XI. Biological Safety Cabinet Malfunction
A failure of a BSC exhausts system is indicated by a red
warning light (insufficient flow) and an alarm. When the alarm sounds, laboratory workers
should follow these procedures:
- Terminate work.
- Cover and contain all vessels containing infectious agents
and contaminated equipment. Turn off all electrical equipment and services, i.e., gas and
vacuum.
- If applicable, close the window completely.
- Notify others in laboratory and leave the room.
- Post signs warning others of the malfunction.
- Call the Biosafety Officer X3560.
- Remain available to provide assistance to the Biosafety
Officer and support personnel.
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XII. Rotor Faillure
If the centrifuge bowl is intact:
- Turn speed control to 0 RPM while letting the vacuum pump
continue to run to remove any aerosol generated.
- Leave the room and warn others not to enter.
- Notify the BSO in order to initiate decontamination
procedures.
- Because of the difficulty of safely and easily introducing
a disinfectant into the centrifuge chamber, paraformaldehyde decontamination of the entire
unit may be required. The BSO will make necessary decontamination arrangements.
- The gaseous decontamination will inactivate aerosolized
particles and small droplets; however, grossly contaminated areas must still be treated as
a biohazards spill. The rotor or rotor fragments will have to be disinfected with a 10%
solution of sodium hypochlorite (household bleach) or Vesphene II se® and
autoclaved.
- The vacuum line and pump will also be contaminated.
Decontamination may require partial dismantling of the centrifuge and pump. The
appropriate centrifuge service-person will be consulted for an acceptable procedure.
If the centrifuge bowl is ruptured:
- This is the equivalent of a biohazard spill in the open
laboratory. Significant amounts of aerosols will be generated. Personnel should leave the
room immediately and warn others not to enter. Contaminated equipment will require
treatment with paraformaldehyde.
- The BSO will determine if the entire room requires gaseous
decontamination. Until decontamination procedures are completed, no one is to enter the
room without proper protective clothing and a full face, HEPA filter respirator.
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XIII.
Gaseous Decontamination
Decontamination vapor may harm the experimental material
or animals. The BSO will advise practical and permissible precautions against vapors.
Gross spills cannot be reliably decontaminated in this way.
Complete decontamination of exposed surfaces in an open
laboratory or cabinet interior can be accomplished with paraformaldehyde. Gaseous
decontamination can be achieved provided that:
- The only possible contamination was by small droplets or
aerosol particles.
- Surfaces were clean before any possible contamination and
remain clean thereafter so that there is maximum contact of the contaminated surfaces with
the paraformaldehyde gas.
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XIV. Chemical
Emergency Management
- If the chemical spill presents an immediate danger,
evacuate all personnel and close the door.
- If spilled material is flammable, turn off ignition and
heat sources.
- Attend to any person who may have been contaminated.
Remove contaminated clothing carefully, folding the contaminated area inward. Place
contaminated clothing in an autoclave bag.
- Thoroughly wash hands and face and any exposed area of the
body. Shower, if necessary, emergency shower/eyewash station is located in the corridor of
the facility and in Room 622.
- Notify the BSO.
- POST SIGNS WARNING OTHERS NOT TO ENTER CONTAMINATED AREA.
- If an airborne infectious agent is involved, make sure a
HEPA filtered respirator is being worn.
- If contamination is aerosolized, leave room and wait one
hour to allow materials to settle before commencing clean up.
- Do not touch the spill without suitable protective
clothing.
- Never assume gases or vapors do not exist or are harmless
because of lack of smell. Review the Material Safety Data Sheet for the chemical to
determine its hazard.
Minor Spills
a. Inform all personnel about the spill, evacuate if
necessary.
- Attend to any person who may have been contaminated.
- Notify the BSO.
- If spilled material is flammable, turn off ignition and
heat sources.
- A chemical "spill kit" is located in each
laboratory and contains equipment for acid, caustic, and organic spills, along with an
instruction booklet.
f. Please read the instruction booklet and be familiar
with the kit before a spill occurs.
In the event that a worker is contaminated by a chemical,
an emergency shower/eyewash station is located in the corridor of the facility.
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XV. Radioactive/Biological Emergency In The Biohazard Facility
Personal Contamination
- Stop work and notify supervisor, BSO, and/or the
Environmental Health and Safety Department.
- If an airborne infectious agent is involved, wear a HEPA
respirator during clean up.
- If clothing or garment is contaminated remove and place in
plastic bag for later handling.
- If body is contaminated wash affected area for at least 15
minutes with mild soap and water and survey with a radiation meter. Use an appropriate
disinfectant during cleaning, if necessary.
- Continue to wash until radiation levels are at background
levels.
- Call the Radiation Safety Officer (RSO) at x2243 for
additional guidance regarding disposal of contaminated materials.
- Contact RSO for guidance regarding autoclaving radioactive
waste.
Area Contamination
- Stop work, evacuate the area and notify the supervisor
and/or Environmental Health and Safety Department.
- If an airborne infectious agent is involved make sure a
HEPA filtered respirator is being worn.
- If contamination is aerosolized, leave room and wait one
hour to allow materials to settle before commencing clean up.
- Wear protective clothing (i.e. Tyvek® suit, gloves,
respirator, etc.) and evaluate the area with a survey meter, if possible. Some radioactive
material cannot be detected by a survey meter, therefore wipe tests should be performed.
- Place absorbent pad over spill area and apply a
disinfectant such as 10% solution of sodium hypochlorite (household bleach) or Vesphene II
se®. Do not use too much disinfectant so that the amount of radioactive waste is limited.
- Allow disinfectant to stand for 30 minutes, this will
provide enough contact time for adequate disinfection.
- Pick up absorbent pad and place it into a plastic bag. If
appropriate, survey area with a meter or take wipe tests to determine if area is free of
radioactive contamination. Continue to clean area with soap and water until the area is
free of contamination.
- Contact the RSO for additional guidance on how to dispose
of your contaminated waste. Autoclaving radioactive waste should be avoided, if possible.
- Wash hands, check shoes, clothing and hands for
contamination with a survey meter after clean up.
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XVI.
Fire and Electrical Failure
Practices to be followed in the event of a fire are as
follows:
1. FIRE INSIDE THE CONTAINMENT FACILITY
- Immediately alert other personnel in the facility to the
danger.
- If possible, without endangering yourself, turn off gas
burners and laboratory equipment and leave the facility immediately using stairwells or
horizontal exits to the Forchheimer Building after notifying other personnel.
DO NOT USE ANY ELEVATORS
- Transmit the alarm. The nearest pull boxes are located
outside the facility next to the stairwell at the south end of the building and at the
bridge to the Forchheimer Building. Follow up with a phone call to x4111.
- If the fire appears containable, (e.g., waste basket or on
a bench top), use the fire extinguishers located throughout the facility. These fire
extinguishers may be used on any type of fire. Be familiar with the operation of the
extinguisher before you have to use it.
TO OPERATE A FIRE EXTINGUISHER
- Remove the unit from the wall and carry or drag it to the
fire area.
- Pull pin.
- Aim nozzle at base of the fire.
- Squeeze handle.
- Sweep nozzle from side to side and gradually progress
forward as the flames are extinguished.
- Once in a position of reasonable safety, notify the
BSO/HAZMAT team concerning any biohazards that have been left exposed. This is the
responsibility of all personnel who are in the facility at the time of the fire.
- Fire Department or other personnel will wear
self-contained breathing apparatus when entrance into the facility is necessary under
emergency conditions. Upon resolution of the emergency, the BSO/HAZMAT team shall
determine if decontamination procedures are required for Fire Department protective
clothing and equipment. Prior to resumption of work in the facility, the BSO shall ensure
that all systems of the facility are in proper operation mode.
2. FIRE IN ANOTHER PART OF THE BUILDING
- If an alarm sounds indicating fire in another part of the
building, personnel should, if possible, turn off gas burners and laboratory-type
equipment.
- Infectious materials should be placed in an incubator,
refrigerator, or freezer. Determine the location of the fire by referring to the chart by
the alarm box.
- Leave the facility, if required, by the most direct route
after notifying other personnel. We encourage evacuation if a fire is reported on the
floor you on which you are working or the floor below. You must evacuate when the
mandatory evacuation alarm sounds.
DO NOT USE ANY ELEVATORS
3. EMERGENCY EVACUATION
Building evacuation may be necessary in certain emergency
situations. The 4-4-4 alarm sequence will activate only in extreme emergencies indicating
that the building must be evacuated immediately by emergency stairwells or horizontal
exits.
ELECTRICAL FAILURE
In the event of power failure in the Biohazard Facility,
all electrical power will be lost for 10 to 15 seconds until the emergency generator is
activated. At this time, only those lights and receptacles on the emergency electrical
power supply, and the Biological Safety Cabinets will be reactivated. The environmental
room (653) and area ventilation systems will be reset manually by engineering personnel.
Power will be lost to all pieces of equipment not connected to the emergency supply. Once
a power failure has occurred, individuals should stop work, decontaminate surfaces, bag or
containerize contaminated items, store cultures safely, and in general, "secure"
the area and leave. Be sure that all doors are closed when exiting.
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XVII.
Operational Procedures
- Only supplies and equipment related to the experiment or
studies shall be introduced into the facility.
- Supplies, equipment, etc., shall not be removed from the
facility unless they have been sterilized or decontaminated under the supervision of the
BSO.
- In order to maintain the established, negative airflow
pattern within the facility, all doors shall be kept closed.
- Use of hypodermic needles and syringes should be limited
to operations or procedures for which there are no alternatives. All sharps should be
disposed of in a sharps container. SHARPS SHOULD NOT BE USED IN HIV LABORATORIES.
- Safe transportation of infectious materials within the
facility requires the use of a secured, labeled non-breakable secondary container.
- All activities involving infectious materials are to be
conducted in a BSC. Handling these agents on the open bench is NOT permitted.
- Mouth pipetting is not allowed. Appropriate pipettes and
pipetting aids are to be provided by the investigator.
- Work surfaces shall be decontaminated daily and
immediately following spills of biohazardous agents with Vesphene II se® or a
designated disinfectant followed by rinsing with distilled water.
- In all procedures, care should be taken to minimize the
creation of aerosols. Any aerosol generating procedure must be performed in a BSC.
- All flasks, test tubes, etc., in which biological agents
are grown or stored shall be appropriately covered to contain potential spills.
- To protect the house vacuum system, all vacuum lines
within the facility must be fitted with an in-line HEPA filter in addition to a secondary
liquid disinfectant trap for biological agents.
- Primary suction flasks must contain appropriate liquid
disinfectant (i.e., 10% household bleach) before use.
- A separate solvent trap is required to capture chemicals,
these chemicals are then properly disposed of under the supervision of the BSO.
- Sinks within this facility are primarily a water source
and hand washing station, they are NOT a disposal area. Contaminated liquids are
not to be poured into the sinks. All waste including tissue culture media, cultures,
buffers, etc., must be autoclaved prior to disposal.
- Due to the communal nature of the facility, all materials
have the potential of being a hazard and should be treated with appropriate caution.
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CENTRIFUGATION - PROCEDURES
All centrifugation must be done in capped centrifuge tubes
and bottles within sealed centrifuge rotors. All microcentrifuges are to be used within a
Biological Safety Cabinet. The following procedure should be followed at all times.
- Review the owners manual.
- Log in name, speed, time and rotor in the appropriate log
book.
- Before centrifuging, inspect tubes, bottles and rotors for
rough spots, pitting, discoloration or cracks.
- Particular care should be given to insuring a proper seal
when using high-speed rotors.
- Make sure the proper adapters are in place. Remove and
disinfect all adapters at the end of a run.
- Fill and decant all centrifuge tubes and bottles within
the Biological Safety Cabinet. Wipe the outside of tubes with disinfectant before placing
in rotor.
- Never overfill centrifuge tubes. Leakage invariably occurs
when tubes are filled to capacity. For general purpose centrifuges, the maximum capacity
should be 3/4 full. Fill ultracentrifuge tubes per manufacturers specifications.
- Balance centrifuge tubes and bottles carefully and wipe
the balance pans with disinfectant after use.
- Never spin uncapped tubes.
- Centrifuge speeds are never to exceed the lower speed
rating for the rotor and/or the test tube used.
- Rotors and/or safety buckets must be opened in a
biological safety cabinet. Wipe the outside surfaces with disinfectant before removing the
rotors from the cabinet.
- Wipe the centrifuge chamber with disinfectant before and
after each spin.
- Decontaminate the rotors and/or buckets after use, by
wiping it with Vesphene II se®. Rinse thoroughly with distilled water when
done, to avoid corrosion.
- Store rotors and accessories in appropriately labeled
cabinets and containers.
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ENVIRONMENTAL SHAKERS - PROCEDURES
- Contents of all vessels must be identified with organism,
user, and telephone extension.
- All vessels containing living organisms must be capped.
These vessels are to be manipulated in such a manner as to prevent wetting of the plugs or
caps.
- Culture flasks or bottles should be held securely in
place, to prevent breakage.
- The shaker is to be kept closed when in motion.
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WATER BATH - PROCEDURES
- Water baths must contain an appropriate disinfectant with
water being changed at reasonable intervals.
- Water baths are to be covered when used for infectious
agents. Appropriate warning signs are to be affixed.
- All vessels containing viable organisms must be closed and
properly labeled.
- Avoid using glassware whenever possible.
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CO2 INCUBATORS - PROCEDURES
- Biological organisms are to be listed on the door of the
incubator identified with the researchers name and extension.
- Humidity reservoirs (where necessary) are to be refilled
with distilled water.
- Door gaskets are susceptible to mold contamination,
therefore, occasionally scrub gaskets with an abrasive cleanser (i.e., BonAmi), rinse with
distilled water, and dry thoroughly.
- Virulent fluid cultures and viable biohazardous materials
should be incubated in non-breakable vessels. Transportation of cultures from the
incubator to the hood shall be in covered, non-breakable, leak-proof pans, trays, or
containers large enough to contain cell fluid in case of leakage.
- Petri plates or other inoculated solid media should be
transported as above.
- Minimize culture contamination, by disinfecting the
incubator with Vesphene II se® and distilled water.
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BLENDERS, HOMOGENIZERS, GRINDERS AND SONICATORS
WHEN WORKING WITH INFECTIOUS AGENTS - PROCEDURES
The equipment listed above creates aerosols when
processing materials. When working with hazardous agents, operate and open the above
equipment in a biological safety cabinet. To reduce the amount of aerosol generated, wait
30 seconds or more after the equipment has been turned off before opening and removing the
processed material. Disinfect all equipment when procedures are complete.
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LYOPHILIZERS - PROCEDURES
The preparation, handling, and use of lyophilized
microorganisms presents an unusual hazard since the accidental release of such powders can
result in aerosols of high concentration. Cross contamination can readily occur,
therefore, handle the powders in a ventilated cabinet. Heavy contamination can also occur
at the air exhaust port or manifold outlets of a lyophilizer. Always wear protective
equipment when removing materials from the lyophilizer.
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XVIII.Waste Disposal-Procedures
Each authorized user is responsible for the autoclaving of
his/her own waste materials, and subsequent removal from the autoclave. Scheduling is
arranged among users.
General Procedures
- Solid waste containers are to be clearly labeled
"Infectious" and lined with two autoclave bags. Waste materials are placed in
the autoclave via the "dirty side" and removed through the "clean
side". The user must log in each autoclave use. When autoclaving is complete the user
is responsible for removing the waste from the autoclave, and discarding it in the red
barrels provided. Our typical recommended autoclaving cycle is 1 hour at 250oC.
- If the autoclave is in use, or malfunctions, return the
waste material to the research laboratory. DO NOT leave the untreated waste on the floor
of the corridor, or in the autoclave room. Always check the autoclave to make sure the
cycle has started properly. Return promptly when the cycle is complete and remove your
waste from the autoclave.
- Pipettes and pipette tips are to be placed in containers
with sufficient disinfectant, to allow disinfectant contact with the entire pipette or
tip. Liquid disinfectants are to be used only for interim decontamination of items. Under
no condition does this constitute a final procedure. Sharps disposal containers must
contain sufficient appropriate disinfectant.
Liquid Waste
- Liquid waste should be mixed with disinfectant (e.g., 10%
household bleach) in the biological safety cabinet, whenever possible.
- After autoclaving or chemical disinfection, liquid wastes
may be carefully poured down the sink with the approval of the Biosafety Officer.
- Never autoclave liquids in non-autoclave containers.
- Never autoclave household bleach or other
chemicals.
- Never autoclave radioactive waste.
- Consult the Biosafety Officer for disposal techniques
concerning hybrid waste material (e.g., radioactive/biohazardous waste).
Solid Waste
- Media bottles, culture flasks, culture tubes, and any
other vessels that may have come in contact with potentially infectious material must be
decontaminated in the biological safety cabinet with sufficient disinfectant before being
placed in biohazard bags and autoclaved.
- Contaminated pipettes may be soaked in sufficient
disinfectant for at least 30 minutes before autoclaving.
- A disinfectant may be used in sharps containers but note
that all sharps containers will be autoclaved before disposal.
Ordinary Waste
- Limit the amount of ordinary trash (e.g., paper,
cardboard), brought into the BHF by removing supplies from their outer packaging
prior to stocking the laboratory.
- Ordinary trash (paper, wrappers, and cardboard is placed
in the hallway for removal by the Housekeeping Staff within the BHF. All ordinary trash
that may be contaminated, shall be autoclaved prior to exiting the facility.
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XIX. Facility
Operations:
The following basic principles underlie the procedures for
movement of personnel and equipment when the containment facility is in use:
- Within the BHF, appropriate protective clothing is always
worn.
- Protective clothing is to be removed when leaving the
containment facility.
- Equipment and material is "autoclave out" except
for items unsuitable for autoclaving.
- Alternative methods of decontamination are determined by
the BSO.
ADMISSION TO THE FACILITY
- Authorized user form must be completed.
- Worker must meet with the BSO.
- Persons falling under any medical restrictions must
satisfy the appropriate requirements.
B. PERSONNEL ACCESS AND EGRESS
1. Regulations for Access Corridor
- Disposable lab wear is to be changed weekly or as
needed.
- Shoe covers and head covers should be used when necessary.
- When required by the BSO, maintenance personnel are to use
disposable Tyvek® suits over street clothes. These suits are placed in a biohazard bag
when exiting the facility for appropriate autoclaving and disposal.
2. Personnel Entering
- Enter airlock.
- Store personal items in assigned lockers, if available.
- Non-disposable cloth lab coats, as well as other
laboratory attire, must not enter the BHF, lab coats may be hung on hooks, provided in the
interlocks.
- Proceed to your work area in street clothing.
- Don personal protective equipment which may typically
consist of: disposable lab coat or Tyvek® suit, shoe covers, head covers and
double latex gloves.
3. Personnel Exiting
- Remove outer pair of gloves when work is completed in
biosafety cabinet.
- Remove disposable lab coat or Tyvek® suit, place it in
infectious waste container or hang it on a coat hook in the facility laboratory.
- Remove second pair of gloves.
- Wash hands thoroughly before exiting.
- Proceed to the corridor in street clothing and exit
facility.
- NO LABORATORY ATTIRE IS TO EXIT THE BHF.
4. Emergency Exit
In an emergency, personnel in the laboratories may leave
via the access corridor without changing clothes, if the situation is significantly
dangerous.
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XX. Access Procedures For Engineering and Other Support Staff:
Maintenance including routine servicing, repair, and
support services will be administered by the BSO. When the laboratory is in use, the
laboratory staff will undertake certain items of routine servicing and support. This
procedure will minimize the necessity for others to enter a potentially contaminated area.
If repairs or replacements in potentially contaminated area are not within the competence
of the laboratory staff, conditions for entry of the maintenance staff and requirements
for decontamination will be defined by the BSO.
- The BSO must be notified at x3560 prior to entry into the
BHF.
- The BSO will approve all entry into the BHF, persons who
comply with all entry and exit procedures will be permitted access to BSL 2 and 3
laboratories or animal rooms.
- All workers will be accompanied by a BSO and be provided
with suitable protective clothing (e.g., disposable lab coat or Tyvek® suit, shoe covers,
head covers, gloves, face mask, etc. as needed). The BSO will determine the appropriate
personal protective equipment for each job.
- If entry is required into a BSL 2 or BSL 3 lab, all
infectious work will be suspended and the area will be declared safe prior to entry.
- Work will be performed under the supervision of the BSO.
- Upon exiting the lab, protective clothing will be removed
and disposed of in the internal change room or corridor.
- Hands will be washed thoroughly before exiting the
laboratory.
- All potentially contaminated tools will be decontaminated
by the BSO before exiting the lab.
* Eating, drinking, smoking, handling contact lenses, and
applying cosmetics are not permitted in the BHF. Persons who must wear contact lenses in
laboratories should also wear goggles or a face shield.
* Persons who have immune dysfunction maybe at risk of
acquiring infections. Persons who are at increased risk of acquiring infection or for whom
infection may be unusually hazardous are not permitted in the BHF laboratories or animal
rooms.
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XXI. Access Procedures For Equipment,and Materials:
GENERAL
Procedures are basically the same for laboratory equipment
(i.e., instruments, glassware), and materials (i.e., media, cultures), all of which are
referred to here as "materials" unless there is a special distinction.
MATERIAL INBOUND
The general rule is that no processing is required for
inbound materials. Such materials may be introduced via the pathway used by inbound
personnel. Special care should be taken when transporting infectious or hazardous
materials into the facility. Overpacking the hazardous material with the addition of an
absorbent is recommended when transporting infectious or hazardous materials.
MATERIAL OUTBOUND
From laboratories
- All materials exiting from the Biohazard Facility must be
either in a closed, plastic autoclave bag, closed buckets or containers. Disposable items
and trash can be placed in double autoclave plastic bags.
- All materials leaving the laboratories are then taken
directly to the autoclave for decontamination. Do not leave materials around the
laboratory for clean up the following day.
- From the Containment Facility
All materials leaving the Containment Facility must be
appropriately disinfected either by surface decontamination, steam sterilization or
formaldehyde decontamination. Large objects may be surface decontaminated within the
laboratory. Removal through the air lock is then possible. Equipment such as microscopes,
water baths, incubators, microcentrifuges, etc., may require formaldehyde decontamination.
This can be arranged with the BSO.
Laboratory Staff
The laboratory staff is to perform routine laboratory
housekeeping, such as autoclaving of lab waste, cleaning of the autoclave chamber,
cleaning of laboratory equipment, and other such actions which constitute good laboratory
practices.
Support Services
The BSO shall arrange for any necessary support services.
These services include service and repair of equipment. Repair of equipment not under
service contract will be made by:
- BSL-2 and 3 personnel
- Albert Einstein College Of Medicine staff
- Outside repair personnel
All major equipment malfunctions must be reported to the
BSO.
Testing and certification of the general air HEPA
filtration systems is performed annually. Testing and certification of the Class II and
III Biological Safety Cabinets within the facility, is to be performed semi-annually or
whenever a cabinet is removed. All major services and repairs must be coordinated through
the BSO. Investigators are responsible for paying the cost of their cabinet certification.
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XXII.
Laboratory Regulations:
All safety and operating procedures applied within each
laboratory fall under the direct responsibility of the investigator and must be in
agreement with the general defined policies of this manual.
Proper recording and control of equipment and safety
operations of the entire BHF are done via a series of sign-up sheets. Records are kept
for:
- All centrifuges in Rooms 622, 623, 624, and 625 (records
are kept on top of equipment).
- Registration of outside persons authorized to enter the
BHF (kept with the BSO).
- Certification and testing reports on airflow, HEPA
filters, sterilizers, safety cabinets (kept with the BSO).
- Pass-through autoclave, users, sterilization checks,
temperature chart records (kept with the BSO).
REGULATIONS FOR AUTOCLAVE USE
- The pass-through autoclave is for decontamination purposes
only.
- Everyone assigned to the BHF must know the operating
procedures of the autoclaves. These procedures are posted on the loading side of the
autoclave.
- All autoclavable material shall leave the laboratory in
covered, sealed, and marked containers.
- Appropriate materials leaving the BHF, whether to be
reused or disposed of, must go through an autoclave decontamination cycle.
- Personnel using the autoclave are to complete a sign-up
sheet, providing name, laboratory, and extension before operating the autoclave.
- The contact of steam with the infectious agent is
essential for any decontamination procedure to work properly.
AUTOCLAVE PROCEDURES
- The screen in the autoclave drain line at the base of the
autoclave is to be removed and cleaned before each use.
- At all times, materials must be placed directly into the
autoclave and not left outside the autoclave.
- Shallow autoclave pans are available for liquid waste
containers and reusable items. Separate pans for each are recommended.
- Uncover or loosen all tightly closed pans, bottles, and
containers when autoclaving.
- Double autoclave biohazard waste bags containing waste are
to be securely closed individually, before leaving the laboratory to be autoclaved.
- Sharps containers are to be covered, placed in double
biohazard bags, and autoclaved as indicated for solid waste.
- After placing a barrel in the autoclave, remove top to
allow steam penetration. The top can also be autoclaved at the same time.
- Barrels are to be relined with double autoclave bags
before being returned to lab.
- Liquid disinfectants are to be used only for interim
soaking of items, before autoclaving. Under no condition does this constitute a final
procedure.
- The BSO will periodically check the efficiency of the
autoclave by testing it with a biological indicator.
- It is the responsibility of the laboratory personnel or
investigator using the autoclave to, load his/her own material, remove it upon completion,
and place the autoclaved waste in the red biohazard containers for removal by the
custodial staff.
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Housekeeping is to be done daily by those authorized users
assigned to the laboratories. Specific scheduling is to be established by the authorized
laboratory personnel.
Room Cleanliness
Each investigator and authorized user is responsible for
his assigned area. Each laboratory must be kept neat and clean. To help facilitate this,
only limited amounts of material needed may be introduced into the facility. No cartons
are permitted. Cleanliness and care within common spaces should be observed by everyone.
Biological Safety Cabinet Cleanliness
Particular attention to cleanliness and tidiness is
necessary within the BSC, to permit convenient and uncontaminated laboratory
manipulations. Equipment and materials should be limited to essential and current needs.
Interior surfaces must be wiped down with Vesphene II se® or other
disinfectant before and after using the cabinet. Attention to interior and exterior of the
windows, is necessary to maximize visibility of manipulations.
Tidiness
To prevent spills and to permit frequent cleaning, a
minimum number of items should be kept on working surfaces.
Pest Control
An important part of housekeeping in biohazard
containment is control of insects. The BSO will coordinate the establishment of this
program.
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XXIV.
Protocol For Custodial Staff:
Admittance of the custodial staff into the BHF
laboratories are arranged with individual laboratories.
- Authorized custodial personnel shall carry out duties in
laboratory rooms before research activities have begun, and/or after research activities
have ceased.
- Appropriate protective clothing (i.e., laboratory coat and
disposable gloves) will be provided and will only be worn in the BHF laboratory.
- The custodial staff shall be responsible for floor, wall
care and keeping the environment of the facility as dust-free as possible. All dusting
will be achieved with a germicide-soaked cloth. This includes removing dust from the
higher reaches of the lab, i.e., on tops of conduits, as well as window ledges, etc.
- Only use treated dry-dust mop heads to suppress the
aerosolization while cleaning.
- Sanimaster Phenolic, a detergent decontamination solution
will be used to wet mop floors and walls.
- Used wet mop heads are to be replaced as needed and are to
be autoclaved before discarding. When autoclaving, the autoclave bag should not be tied to
allow the steam to penetrate.
- Other responsibilities include filling paper towel
dispensers, iodoform soap dispensers, washing all door handles with germicidal solutions,
and washing the corridor floors with Sanimaster Phenolic.
- None of the custodial equipment is to be left in the
corridor, all supplies should be returned to the closet.
- Thorough floor cleaning, i.e., scrubbing and stripping is
to be done as needed. Scheduling is to be coordinated with the BSO.
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XXV. Laboratory
Storage:
Laboratory storage capacity is limited, therefore, only a
limited amount of material needed may be introduced into the facility. No cartons are
permitted to be stored on the floor.
All infectious materials requiring BSL-2 or 3 containment
must be stored in closed, sealed containers, and their contents and location identified
appropriately by biohazard sign and the name of the researcher, department, room, and
extension.
Environmental Room/Cold Room
- Space will be assigned by the BSO to the researcher.
- It is expected that each researcher will remove materials
and decontaminate the area promptly upon the completion of the project within the
facility.
Freezer and Refrigerator Space
To be assigned by the BSO.
- Prompt removal and proper disposal of materials is
expected upon completion of experimental protocols and projects.
- Investigators should not expect to be able to use storage
space longer than 6 months after completion of their project.
- Careful storage of biological materials is essential.
Storage of minimum amounts of materials is expected.
- Flammables are not permitted to be stored in non-explosion
proof refrigerators or freezers.
Chemical Storage
- No more than five gallons of flammable liquids per
laboratory is allowed.
- Acids must be stored separately from solvents.
- Store large bottles as close to the floor as possible, but
NO chemicals may be stored on the floors or in the aisles.
- Storage is not permitted in corridors.
- Isotopes must not be stored in the BSL-2 or 3 facility.
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XXVI.
Disinfectant Solutions:
For surface decontamination (one of the following may be
used):
- Vesphene II se®
- Wescodyne, 2% final concentration
- Clorox, (household bleach), 10% final concentration
For liquid wastes containing viruses, virus-infected or
transformed cells:
- Vesphene II se®
- Wescodyne, 2% final concentration
- Clorox, (household bleach), 10% final concentration
Pipette decontamination:
- Clorox, (household bleach), 10% final concentration
- Decontamination of overt spills of biohazardous material:
- Vesphene II se®
- Clorox, (household bleach), 10% final concentration
Floors:
- Sanimaster Phenolic Cleaning Detergent
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XXVII. Authorized
User:
Requirements for BHF personnel:
- Complete medical requirements.
- Study the Albert Einstein College of Medicine Biohazard
Facility Safety and Operations Manual and other guidelines where applicable.
- Receive training in the use of the facilitys
containment equipment and proper techniques.
- Demonstrate familiarity with facility procedures.
- Sign the statement below. Review the Containment
Facilitys Safety and Operations Manual semiannually and re-initial this statement.
- Receive appropriate keys and locker assignment from BSO.
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XXVIII.
Containment Facility Keys:
I understand that the Containment Facility keys issued to
me are:
- my responsibility
- not, under any circumstances, to be shared, distributed,
given on loan and/or reproduced.
- to be returned to the Biosafety Officer when no longer
needed by me.
- If any Containment Facility keys issued to me are lost, I
will notify the Biosafety Officer immediately.
- I have read and understand the information contained in
the AECOM Biohazard Facility Safety and Operations Manual. I understand my
responsibilities as an authorized user of the Facility.
______________
_______________________________________________
Date
Signature
______________
_______________________________________________
Date
Biosafety Officer
_______________
_______________________________________________
Date
Principal Investigator or Designee
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XXIX. Autocave (Room 675 Chanin Building) - Biohazard Facility:
OPERATING INSTRUCTIONS FOR
AUTOCLAVE (STEAM CYCLE)
- Complete sign-up sheet
- Remove debris from chamber as needed(check strainer-clean before each ude)
- Master power switch - "ON".
- Check that clean side door is locked.
- Check jacket pressure gauge:it should read 15 to 20 pound
- Liquid cycle button should be depressed (Open door and clean strrainer located on the bottom surface of the autoclave
- Place contaminated materials in chamber.
- Check timer-60 minutes.
- Lock door recheck jacket pressure.
- Press "cycle start" button.(sterilizer then goes through sequence automatically)(buzzer will sound on clean side when cycle is complete)
- Open clean side door and removel material.
- Lock clean side door.
- Open contaminated side door to relieve gasket pressure.
- Close contaminated side door.
AUTOCLAVE IS NOW READY FOR ANOTHER CYCLE.
DO NOT USE DRY CYCLE.
DO NOT PRESS THE "CYCLE OFF" BUTTON.
DO NOT LEAVE DOOR ON CONTAMINATED SIDE LOCKED WHEN NOT IN
USE.
ANY QUESTIONS OR PROBLEMS, CALL X3560
*This step is critical. If strainer is clogged, the steam
cannot be exhausted from the autoclave.
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XXX. GENERIC
DISINFECTANT CHART:
ASSESSMENT OF GENERIC DISINFECTANTS ON BASIS OF EFFICACY
| DISINFECTANT |
CIDAL
ACTIVITY INDICATED |
| Class |
Bacteri- |
Tuberculo- |
Pseudomona- |
Sport- |
Viru- |
| Acids/alkalies |
good |
good |
good |
good |
good |
| Alcohols |
good |
good |
good |
none |
moderate |
| Chlorines |
good |
good |
good |
moderate |
good |
| Formaldehyde |
good |
good |
good |
good |
good |
| Glutaraldehyde |
good |
good |
good |
good |
good |
| Iodines |
good |
good |
good |
moderate |
good |
| Mercurials |
fair |
none |
fair |
none |
good |
| Phenolics |
good |
good |
good |
poor |
moderate |
| Quaternaries |
good |
none |
fair |
none |
moderate |
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