Research Compliance/EHS | 2015
BIOSAFETY MANUAL
PI Name
Table of Contents
I.
Introduction to Biosafety
II.
Hazard Identification/Mitigation
III.
Labels and Signs
IV.
Decontamination
V.
Waste Disposal
VI.
Transport/Shipping
VII.
Emergency Response
VIII. Lab Map
IX.
IBC Application
X.
Exposure Control Plan (Bloodborne Pathogens: Human Materials)
Appendix
A. Example of a Pathogen Safety Data Sheet
B. Example of Hepatitis B Vaccine Declination Form
C. Example of a Sharps Injury Log
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Directions:
1. Provide the information as noted in the specific sections;
2. In addition, if your research involves the use of human/nonhuman primate
materials including cell lines, fill out the Exposure Control Plan section;
3. Place the manual in a binder and make readily accessible for all lab
personnel;
4. Have all lab personnel read and sign using the signatory form at the end of
the document.
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I. Introduction
The fundamental objective of a biosafety program is the containment of potentially harmful
biological material. “Containment” is used to describe the safe methods, facilities and
equipment for managing biohazardous material in the laboratory or animal facility. The
purpose of containment is to reduce or eliminate exposure of personnel and the environment
to this material. Biological containment is designated by 4 biosafety levels in ascending order
(1 through 4) characterized by increasing levels of protection provided to personnel, the
environment and the community.
This manual identifies the hazards that may be encountered in this lab/facility and specifies
practices and procedures designed to minimize or eliminate exposure.
Routes of Transmission
There are 5 main routes of transmission of biohazardous materials in the laboratory/animal
facility.
1. Inhalation of aerosols: Many lab procedures can cause the aerosolization of
biohazardous material. Some of these procedures include the use of vortexers,
blenders, sonicators and centrifuges. Proper work practices and engineering controls
must be implemented to minimize aerosolization especially of organisms whose normal
route of transmission is by the aerosol route (e.g. adenovirus, Mycobacterium
tuberculosis, etc.) Aerosolization can also result in environmental contamination of the
work area.
2. Ingestion: Accidental ingestion can result from improper personal hygiene such as
inadequate/improper hand washing. Food and drink are prohibited in all areas of the lab
in which work is conducted with potentially biohazardous material due to the risk of
contamination.
3. Spill/Splash: Exposures to mucous membranes and/or eyes can result from aerosols or
splashes to the eyes, nose and/or mouth.
4. Percutaneous: These injuries are caused by any kind of contaminated sharp (needles,
broken glass, necropsy scissors, etc.) piercing the skin. This type of injury is particularly
serious because of direct inoculation into the normally sterile bloodstream.
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5. Indirect: Ingestion, mucous membrane and/or eye exposure from inadvertent inoculation
by contaminated hands or contaminated items such as cell phones, ear buds, pens, etc.
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II. Hazard Identification and Control
BACKGROUND: There are many hazards present in a laboratory including physical,
mechanical, chemical, electrical and biological. This manual specifically addresses biological
hazards but may be adapted to incorporate all hazards.
Biohazards can be any of the following:
1. biological entities or products/parts of biological entities (e.g. toxins) including recombinant
or synthetic nucleic acid molecules that fall under the NIH Guidelines for Research
Involving Recombinant or Synthetic Nucleic Acid Molecules
2. equipment in contact with biologic entities/products
3. procedures/operations involving biologic entities/products
4. poor behaviors or practices (such as not washing hands before leaving lab).
Biohazards: In order to eliminate or mitigate the risk of exposure and to educate lab personnel
on the risks, it is important to first identify the most likely lab routes of exposure, the signs and
symptoms of infection/intoxication, and the range of outcomes that may result from infection or
intoxication (subclinical to death).
Equipment: Proper use of laboratory equipment is required to work safely with hazardous
materials such as chemicals and biohazards. Training on the correct use, maintenance and
emergency response in the event of failure are essential for preventing accidents and possible
exposure. Examples of equipment that have been implicated in laboratory-acquired infections
include centrifuges, vortexers, homogenizers, sonicators, plate washers, autoclaves, shaking
incubators and other pieces capable of producing aerosols or splashes.
Lab Procedures/Operations: Manipulation of the biohazard can result in exposure via any of
the 4 routes listed in Section I. Clearly written and validated SOPs are critical to ensure safety
and the generation of sound research data. Hazardous procedures include anything that can
generate an aerosol or splash as well as use of sharps. Culturing, plating, and harvesting of
microorganisms are considered hazardous especially if done on the lab bench. Animal
handling and inoculation activities are also increase risk of exposure to hazards.
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Lab Practices: Standard and special microbiological practices are described in Section IV of
the CDC’s Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as
descriptions of the four biosafety levels. Proper hand washing is the single most important
infection control procedure that can be utilized in a lab or animal facility. Wash hands
after removing gloves and before leaving the lab! Eating, drinking and any other
manipulation around the head area is prohibited.
HAZARD IDENTIFICATION AND CONTROL
Identify all biohazardous material, hazardous operations and equipment present in this
laboratory/facility and describe how the risk of exposure will be eliminated or mitigated by the
use of the hierarchy of controls [engineering, work practice and personal protective equipment
(PPE)].
1. List all potentially biohazardous entities or products/parts including hazardous
recombinant or synthetic nucleic acid material. Attach the Canadian Public Health
Service Pathogen Safety Data Sheets (PSDS) for each organism if available.
http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php
If a PSDS is not available for your biohazard, then use as a template to collect
information to create your own PSDS. Be sure to include the most likely routes of
transmission, signs and symptoms, and the range of outcomes from
infection/intoxication. An example of a PSDS is provided in Appendix A.
[INSERT]
2. List all potentially hazardous equipment (aerosol or splash generating) that will be used
and describe what safety measures will be implemented using the hierarchy of controls
(engineering, work practice, PPE).
[INSERT]
3. List all potentially hazardous procedures/operations (aerosol or splash generating,
sharps use) and describe what safety measures should be followed using the hierarchy
of controls (engineering, work practice, PPE).
[INSERT]
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4. The standard and special microbiological practices from the BMBL as well as the
description of the biosafety level for the facility are included here as basic information.
Practices specific for your lab should be described in sections 1 through 3 on the
preceding pages.
Biosafety Level 2
Biosafety Level 2 builds upon BSL-1. BSL-2 is suitable for work involving agents that pose
moderate hazards to personnel and the environment. It differs from BSL-1 in that: 1)
laboratory personnel have specific training in handling pathogenic agents and are
supervised by scientists competent in handling infectious agents and associated
procedures; 2) access to the laboratory is restricted when work is being conducted; and 3)
all procedures in which infectious aerosols or splashes may be created are conducted in
BSCs or other physical containment equipment.
The following standard and special practices, safety equipment, and facility requirements
apply to BSL-2.
A. Standard Microbiological Practices (Applies to all 4 biosafety levels)
1. The laboratory supervisor must enforce the institutional policies that control access to the
laboratory.
2. Persons must wash their hands after working with potentially hazardous materials and
before leaving the laboratory.
3. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food
for human consumption must not be permitted in laboratory areas. Food must be stored
outside the laboratory area in cabinets or refrigerators designated and used for this
purpose.
4. Mouth pipetting is prohibited; mechanical pipetting devices must be used.
5. Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken
glassware must be developed and implemented. Whenever practical, laboratory
supervisors should adopt improved engineering and work practice controls that reduce risk
of sharps injuries. Precautions, including those listed below, must always be taken with
sharp items. These include:
a. Careful management of needles and other sharps are of primary importance.
Needles must not be bent, sheared, broken, recapped, removed from disposable
syringes, or otherwise manipulated by hand before disposal.
b. Used disposable needles and syringes must be carefully placed in conveniently
located puncture-resistant containers used for sharps disposal.
c. Non-disposable sharps must be placed in a hard walled container for transport to a
processing area for decontamination, preferably by autoclaving.
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d. Broken glassware must not be handled directly. Instead, it must be removed using a
brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware
whenever possible.
6. Perform all procedures to minimize the creation of splashes and/or aerosols.
7. Decontaminate work surfaces after completion of work and after any spill or splash of
potentially infectious material with appropriate disinfectant.
8. Decontaminate all cultures, stocks, and other potentially infectious materials before
disposal using an effective method. Depending on where the decontamination will be
performed, the following methods should be used prior to transport:
a. Materials to be decontaminated outside of the immediate laboratory must be placed
in a durable, leak proof container and secured for transport.
b. Materials to be removed from the facility for decontamination must be packed in
accordance with applicable local, state, and federal regulations.
9. A sign incorporating the universal biohazard symbol must be posted at the entrance to
the laboratory when infectious agents are present. Posted information must include: the
laboratory’s biosafety level, the supervisor’s name (or other responsible personnel),
telephone number, and required procedures for entering and exiting the laboratory. Agent
information should be posted in accordance with the institutional policy.
10. An effective integrated pest management program is required. (See Appendix G.)
11. The laboratory supervisor must ensure that laboratory personnel receive appropriate
training regarding their duties, the necessary precautions to prevent exposures, and
exposure evaluation procedures. Personnel must receive annual updates or additional
training when procedural or policy changes occur. Personal health status may impact an
individual’s susceptibility to infection, ability to receive immunizations or prophylactic
interventions. Therefore, all laboratory personnel and particularly women of childbearing
age should be provided with information regarding immune competence and conditions that
may predispose them to infection. Individuals having these conditions should be
encouraged to self-identify to the institution’s healthcare provider for appropriate counseling
and guidance.
B. Special Practices (Specific for BSL-2)
1. All persons entering the laboratory must be advised of the potential hazards and meet
specific entry/exit requirements.
2. Laboratory personnel must be provided medical surveillance, as appropriate, and offered
available immunizations for agents handled or potentially present in the laboratory.
3. Each institution should consider the need for collection and storage of serum samples
from at-risk personnel.
4. A laboratory-specific biosafety manual must be prepared and adopted as policy. The
biosafety manual must be available and accessible.
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5. The laboratory supervisor must ensure that laboratory personnel demonstrate proficiency
in standard and special microbiological practices before working with BSL-2 agents.
6. Potentially infectious materials must be placed in a durable, leak proof container during
collection, handling, processing, storage, or transport within a facility.
7. Laboratory equipment should be routinely decontaminated, as well as, after spills,
splashes, or other potential contamination.
a. Spills involving infectious materials must be contained, decontaminated, and cleaned
up by staff properly trained and equipped to work with infectious material.
b. Equipment must be decontaminated before repair, maintenance, or removal from the
laboratory.
8. Incidents that may result in exposure to infectious materials must be immediately
evaluated and treated according to procedures described in the laboratory biosafety
manual. All such incidents must be reported to the laboratory supervisor. Medical
evaluation, surveillance, and treatment should be provided and appropriate records
maintained.
9. Animal and plants not associated with the work being performed must not be permitted in
the laboratory.
10. All procedures involving the manipulation of infectious materials that may generate an
aerosol should be conducted within a BSC or other physical containment devices.
C. Safety Equipment (Primary Barriers and Personal Protective Equipment)
1. Properly maintained BSCs, other appropriate personal protective equipment, or other
physical containment devices must be used whenever:
a. Procedures with a potential for creating infectious aerosols or splashes are
conducted. These may include pipetting, centrifuging, grinding, blending, shaking,
mixing, sonicating, opening containers of infectious materials, inoculating animals
intranasally, and harvesting infected tissues from animals or eggs.
b. High concentrations or large volumes of infectious agents are used. Such materials
may be centrifuged in the open laboratory using sealed rotor heads or centrifuge safety
cups.
2. Protective laboratory coats, gowns, smocks, or uniforms designated for laboratory use
must be worn while working with hazardous materials. Remove protective clothing before
leaving for non-laboratory areas, e.g., cafeteria, library, and administrative offices). Dispose
of protective clothing appropriately, or deposit it for laundering by the institution. It is
recommended that laboratory clothing not be taken home.
3. Eye and face protection (goggles, mask, face shield or other splatter guard) is used for
anticipated splashes or sprays of infectious or other hazardous materials when the
microorganisms must be handled outside the BSC or containment device. Eye and face
protection must be disposed of with other contaminated laboratory waste or
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decontaminated before reuse. Persons who wear contact lenses in laboratories should also
wear eye protection.
4. Gloves must be worn to protect hands from exposure to hazardous materials. Glove
selection should be based on an appropriate risk assessment. Alternatives to latex gloves
should be available. Gloves must not be worn outside the laboratory. In addition, BSL-2
laboratory workers should:
a. Change gloves when contaminated, glove integrity is compromised, or when
otherwise necessary.
b. Remove gloves and wash hands when work with hazardous materials has been
completed and before leaving the laboratory.
c. Do not wash or reuse disposable gloves. Dispose of used gloves with other
contaminated laboratory waste. Hand washing protocols must be rigorously followed.
5. Eye, face and respiratory protection should be used in rooms containing infected animals
as determined by the risk assessment.
D. Laboratory Facilities (Secondary Barriers)
1. Laboratory doors should be self-closing and have locks in accordance with the
institutional policies.
2. Laboratories must have a sink for hand washing. The sink may be manually, hands-free,
or automatically operated. It should be located near the exit door.
3. The laboratory should be designed so that it can be easily cleaned and decontaminated.
Carpets and rugs in laboratories are not permitted.
4. Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces
between benches, cabinets, and equipment should be accessible for cleaning.
a. Bench tops must be impervious to water and resistant to heat, organic solvents,
acids, alkalis, and other chemicals.
b. Chairs used in laboratory work must be covered with a non-porous material that can
be easily cleaned and decontaminated with appropriate disinfectant.
5. Laboratory windows that open to the exterior are not recommended. However, if a
laboratory does have windows that open to the exterior, they must be fitted with screens.
6. BSCs must be installed so that fluctuations of the room air supply and exhaust do not
interfere with proper operations. BSCs should be located away from doors, windows that
can be opened, heavily traveled laboratory areas, and other possible airflow disruptions.
7. Vacuum lines should be protected with liquid disinfectant traps.
8. An eyewash station must be readily available.
9. There are no specific requirements for ventilation systems. However, planning of new
facilities should consider mechanical ventilation systems that provide an inward flow of air
without recirculation to spaces outside of the laboratory.
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10. HEPA filtered exhaust air from a Class II BSC can be safely recirculation back into the
laboratory environment if the cabinet is tested and certified at least annually and operated
according to manufacturer’s recommendations. BSCs can also be connected to the
laboratory exhaust system by either a thimble (canopy) connection or directly exhausted to
the outside through a hard connection. Provisions to assure proper safety cabinet
performance and air system operation must be verified.
11. A method for decontaminating all laboratory wastes should be available in the facility
(e.g., autoclave, chemical disinfection, incineration, or other validated decontamination
method).
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III. Labels and Signs
A warning label that includes the universal biohazard symbol, followed by the term "biohazard,"
must be included on bags/containers of biohazardous waste, on bags/containers of
contaminated laundry, on equipment that is used for culturing, manipulating or storing
biohazardous material, and on bags/containers used to store, dispose of, transport, or ship
biohazardous material (e.g., specimen containers). Typically, equipment is decontaminated by
lab personnel before service or shipment. However, if contaminated equipment is to be serviced
or shipped, it must have a readily observable label attached which contains the biohazard symbol
and the word "biohazard" along with a statement relating which portions of the equipment remain
contaminated.
Some examples of the universal biohazard symbol:
Other types of signage:
The background must be fluorescent orange or orange-red or predominantly so, with symbols
and lettering in a contrasting color. The label must be either an integral part of the container or
affixed as close as feasible to the container by a string, wire, adhesive, or other method to
prevent its loss or unintentional removal. Red bags or red containers may be substituted for the
biohazard labels. The supervisor or PI is to be notified if items are not appropriately labeled.
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IV. Decontamination
This section should describe the basic strategies for decontaminating surfaces, equipment and
spaces in your facility to eliminate the possibility of transmission to lab personnel, the general
public and the environment.
NOTE: Recombinant and/or synthetic nucleic acid molecules as defined by the NIH Guidelines
for Research Involving Recombinant or Synthetic Nucleic Acid Molecules are considered
biohazardous and must be decontaminated in accordance with the Guidelines. See IBC-SOP001 for guidance.
List the disinfectants that will be used for the decontamination of surfaces and equipment.
Include concentration and contact (kill) time.
[Insert procedure here.]
Describe the procedures for handling dirty and/or contaminated lab coats.
[Insert procedure here.]
If the risk assessment requires the use of reusable PPE (safety goggles, face shields) describe
the procedure for cleaning and decontamination.
[Insert procedure here.]
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V. Waste Disposal
All waste containing potentially biohazardous material must either be sterilized on site or
removed by MSU’s contracted medical waste vendor.
NOTE: Recombinant and/or synthetic nucleic acid molecules as defined by the NIH Guidelines
for Research Involving Recombinant or Synthetic Nucleic Acid Molecules are considered
biohazardous waste and must be disposed of in accordance with the Guidelines. See IBCSOP-001 for guidance.
Steam sterilization using an autoclave is one of the most common methods used to render
material sterile (ALL organisms including spores are destroyed).
Autoclave: Any autoclave used for sterilization of potentially biohazardous material must have
an autoclave verification program. Please contact the biosafety officer for details. Once a load
has been confirmed as sterilized the following MUST occur:
1. Label the autoclaved red bag with a label that says “TREATED WASTE”.
2. Place the now sterile red bag into a black plastic trash bag and seal the black bag.
3. Take the trash bag containing the sterilized red bag to the nearest dumpster and
carefully place inside.
4. Custodians will never handle waste from a laboratory so it is the responsibility of the lab
personnel to dispose of the waste.
Noninfectious Medical Waste: Noninfectious medical waste (IV bags, unused media, unused
agar plates and broth tubes, etc.) must be
1. labeled “NONINFECTIOUS”
2. placed into a black plastic trash bag
3. And taken to the nearest dumpster by lab personnel.
Sharps: DO NOT place any kind of sharps in a bag. All sharps, contaminated or not, must go
into some kind of puncture-proof, leak-proof container. All sharps contaminated with a potential
biohazard must go into a RED sharps container. Close and process for decontamination when
the container is 2/3 full. Red sharps containers can be autoclaved in-house or picked by EHS
for disposal by MSU’s contracted medical waste vendor.
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Untreated Waste: Untreated waste for vendor removal may be picked up by the hazardous
waste officer in the Environmental Health & Safety office. Please submit a request through the
EHS web site.
Liquid Disinfection: Some liquids may be chemically decontaminated. If your lab will be using
chemical decontamination please describe the procedure below. Please note that complete
reliance should not be placed on liquid disinfection if the end result must be sterility.
[Insert procedure here if applicable.]
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VI. Transport/Shipping
Transport of biohazardous material from one place to another whether to the lab next door or
across campus should be done under triple containment to prevent release of the material in
the event of an accident.
Shipping to another entity falls under a number of federal and international regulatory agencies
and requires specific training. Please contact the EHS office for additional information.
Triple containment is described as the primary container holding the material (test tube, agar
plate, etc.,) placed into a leak-proof, shatter-proof secondary container marked with the
biohazard sign which is then placed into another leak-proof, shatter-proof tertiary container.
The outside of all 3 should be decontaminated with 70% ethanol (or other appropriate
disinfectant) before transport and when the transfer is completed.
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VII. Emergency & Incident Response Plan
Use the following procedures for guidance in developing your lab specific emergency
response.
NOTE: Recombinant and/or synthetic nucleic acid molecules as defined by the NIH Guidelines
for Research Involving Recombinant or Synthetic Nucleic Acid Molecules are considered
biohazardous and any accidents or personnel exposures must be handled in accordance with
the Guidelines. This includes reporting certain types of incidents to the NIH Office of
Biotechnology Activities (OBA). See IBC-SOP-001 for guidance.
A. Contact Information:
Name
Contact Numbers
Principal Investigator:
Office:
Cell :
Alternate Contact:
Office:
Cell :
Emergency
911
University Police
662-325-2121
Student Health Center
662-325-2431
Oktibbeha County Hospital
662-323-4320
MSU Campus Facilities
Computer/Phone Problems
662-325-0631
Facilities Management
662-325-2005
EHS
662-325-3294
Research Compliance
662-325-3294
Biological Spill
662-325-3294
Radiation Spill
662-325-3294
Chemical Spill
662-325-3294
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B. Fire or Evacuation Emergency (gas leak, explosion, power outage)
1. Fire - Call 911 -Give name, location and provide information requested. Secure
biohazardous agent by locking the freezer if possible. Do not endanger yourself.
2. Pull fire alarm.
3. Evacuate building – follow exit signs (see evacuation route in Section VIII); go to
rally location.
LIST RALLY POINT HERE.
4. The PI will call role and identify missing to emergency responder.
C. Biological Spill
A MINOR BIOLOGICAL SPILL is one that can be handled safely by laboratory
personnel without the assistance of safety and emergency personnel. Minor spills
include:

The release of BSL-1 organisms without splashing or agitation

The release of a small volume of BSL-2 organisms without splashing or agitation
A MAJOR BIOLOGICAL SPILL is one that requires outside assistance. These include:

The release of BSL-2 organisms resulting in excessive splashing and agitation

The release of a large volume of BSL-2 organisms (there is enough present to
seek its own level or in other words, to run to a low point)
Call 325-3294 or 325-2121 after hours.
Spill Response
Each laboratory should have a Spill Response Plan and a Spill Kit on hand. The Spill
Response Plan should be available to all personnel and contains 4 elements: the use
and availability of appropriate PPE, assessments of the nature and extent of various
spills, the use of appropriate disinfectants, and disposal.
The kit should be maintained in a white 5-gallon leak-proof bucket and contain the
following:
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
Concentrated household bleach (check expiration date) or other appropriate
disinfectant

Spray bottle for making 10% bleach solution

Forceps or tongs for handling sharps

Paper towels or other suitable absorbent

Biohazard bags of various sizes

Disposable gloves

Disposable foot covers

Face protection – at a minimum safety glasses and mask

Disposable apron, gown or tyvek suit
Spill or Splash ton Body
1. Remove contaminated clothing.
2. Gently wash exposed area with soap and water for at least 1 minute.
3. If eye or mucous membrane exposure occurs, use eye wash per instructions.
Flush for approximately 5 minutes.
4. Report spill to supervisor and BSO.
5. Obtain medical attention if necessary.
Inside the Biosafety Cabinet
1. Wait at least 5 minutes to allow BSC to filter aerosols.
2. Wear lab, coat, sleeve guards, safety glasses, and gloves during clean-up. You
may want to double glove in the event the outer pair becomes contaminated.
3. Allow BSC to run during clean-up.
4. Apply disinfectant for a minimum 20 minute contact time (contact time depends
upon the specific disinfectant).
5. Wipe up spill with disinfectant-soaked paper towels or absorbent pillows.
6. Wipe the walls, work surfaces, inside of sash and any equipment with
disinfectant-soaked paper towels.
7. Lift exhaust grill and tray and wipe all surfaces.
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8. Discard contaminated disposable materials using appropriate biohazardous
waste disposal procedures.
9. Wipe down contaminated reusable items with disinfectant then place in
biohazard bags or autoclave pans with lids for autoclaving.
10. Those items that are non-autoclavable should be wiped down with disinfectant
and kept wet for a minimum of 20 minutes before removal from BSC.
11. Remove protective clothing when done and place in biohazard bag for
autoclaving.
12. Run the BSC for 10 minutes after clean-up before reusing.
13. WASH HANDS!
In the Laboratory, Outside of BSC
1. Call the BSO if a major spill.
2. Clear the room of all personnel.
3. Remove any contaminated clothing and place in biohazard bag for autoclaving.
4. Wait at least 30 minutes for aerosols to settle before reentry.
5. Put on either a Tyvek suit or disposable gown, disposable foot covers, gloves,
and safety glasses.
6. Place dry paper towels on the spill then layer a second set of disinfectant-soaked
towels over the spill.
7. Starting from the outside and working in, carefully soak the spill with disinfectant
being careful to minimize aerosolization.
8. Decontaminate all items within the spill area. Wait at least 20 minutes contact
time with the disinfectant (contact time depends upon the specific disinfectant).
9. Wipe equipment and reusable items with the disinfectant.
10. Discard contaminated disposables in biohazard bags.
11. If sharps are present, use a mechanical device such as a dust pan and brush to
pick up the spill and place contaminated sharps in an approved sharps container.
Inside a Centrifuge
1. Clear area of personnel.
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2. Wait at least 30 minutes for aerosols to settle before clean-up.
3. Wear a lab coat, gloves, and safety glasses during clean-up.
4. Wipe rotors and buckets with disinfectant then remove to nearest BSC for more
extensive decontamination.
5. Thoroughly disinfect inside of centrifuge with a minimum contact time of 20
minutes (contact time depends upon the specific disinfectant).
6. Dispose of contaminated materials using appropriate biohazardous waste
disposal procedures.
Outside the Laboratory, In Transit
1. To prevent or minimize a spill, transport materials in triple containment using an
unbreakable, leak-proof, sealed secondary container placed inside a tertiary
unbreakable, leak-proof, sealable container. The secondary container should
labeled with the universal biohazard symbol.
2. Should a spill occur in a public area, do not attempt to clean up without
appropriate PPE.
3. Secure the area around the spill.
4. Call the Biosafety Office 325-0620.
5. Stand by for further assistance if required.
Reporting of Accidents: Major spills and personnel exposure incidents should be
reported by the PI or supervisor to the BSO. The BSO in conjunction with the IBC Chair
will conduct an investigation of the laboratory accident. The goal of the investigation is
to analyze the events surrounding the accident to prevent or minimize its reoccurrence
and to identify those personnel involved in the event further medical surveillance is
needed.
Any accidents or personnel exposures involving recombinant and/or synthetic nucleic
acid molecules as defined by the NIH Guidelines for Research Involving Recombinant
or Synthetic Nucleic Acid Molecules must be handled in accordance with the
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Guidelines. This includes reporting certain types of incidents to the NIH Office of
Biotechnology Activities (OBA). See IBC-SOP-001 for guidance.
(excerpt from the NIH Guidelines: Section IV-B-2-b-(7). Reporting any significant problems
with or violations of the NIH Guidelines and any significant research-related accidents or
illnesses to the appropriate institutional official and NIH/OBA within 30 days. Incidents
occurring under BSL-2 conditions that result in an overt exposure to organisms containing
recombinant or synthetic nucleic acid molecules must be reported to NIH OBA immediately.)
Please report incidents that did not result in an exposure (near miss) to the BSO.
Evaluation of near misses can lead to alternative work practices and implementation of
engineering controls to minimize future incidents.
Sharps Injury: Whenever an injury occurs involving a contaminated sharp (needle,
broken glass, etc.), the BSO must be notified. The subsequent investigation will
determine if a safer device or work practice can be used to reduce or prevent the
accident from reoccurring. Seek medical attention immediately for a contaminated
sharps injury!
D. Chemical Spill
1. Evacuate area if necessary for personal safety
2. Notify other people and supervisor
3. Take action to contain spill if properly trained. Do not endanger yourself.
4. Call-Day (8:00am-5pm) 325-2787 After hours MSU PD 325-2121
E. Radiation Spill
1. Evacuate appropriate area
2. Call Radiation Safety- Day 325-2787 Night 325-2121
3. Take action to contain spill if properly trained. Do not endanger yourself.
F. Medical Emergency
1. If an individual is injured, call 911. Employees are not required to perform first aid.
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2. Notify supervisor
3. Do not move person or perform any first aid if you are not trained and qualified.
G. Animal Bites
1. Clean bite thoroughly with soap and water.
2. Apply bandage
3. Notify supervisor
4. Obtain medical attention.
H. Tornado or Hurricane
1. Building emergency coordinator should have an emergency weather radio.
2. Move away from windows into basement or interior hallway on a lower floor.
3. Avoid auditoriums, gymnasiums, or other areas having a wide, free span roof.
4. Take cover under heavy furniture
5. If outdoors, lie flat in the nearest depression such as a ditch or ravine. If there is
time, move away from the path of the tornado/hurricane.
I. Other natural disasters or emergencies
In the event of some other disaster or emergency requiring evacuation, individuals in an
affected space should evacuate in an orderly manner, closing the door behind them.
They should assemble in a pre-determined gathering place for further instructions. The
University police responders will assist in the evacuation.
J. Mechanical/Facility concerns
1. Do not attempt to repair failure (e.g. burst pipe, elevator stuck, temperature, power
outage, etc.,)
2. Call 325-2005
3. Give name, location, and state problem. Stay on phone until information
acknowledged.
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K. Suspicious Persons Or Activities
Any suspicious persons or activities must be immediately reported to MSU PD at 3252121. Do not open the laboratory door to allow any suspicious or unknown persons
entry to the laboratory.
L . Oral Threat
1. Oral threat spoken or called into building.
2. Record time, date and if you have caller ID note phone number.
3. Have co-worker call on another line to MSU PD 325-2121 or 911.
4. Keep caller on line as long as possible.

Where is incident going to occur?

When is it going to happen?

Listen for background noise

Listen for voice
5. Suspicious letter/package

Suspicious letter or package

Turn off all cell phones and pagers.

Look for no return address, misspelled words in address or return, excessive
postage, stains, or other visible signs.


Do not shake, or move the package.

Isolate letter or package

Do not open or smell

Immediately notify PI, and call MSU PD 325-2121 or 911.
Describe letter or package, give name, location and remain calm.
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VIII. Lab Map
Insert a map of the facility showing placement of safety equipment including eye
wash, shower, fire extinguisher, BSC, spill kit and escape route.
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IX. IBC Application
Insert a copy of the IBC application here. This document describes the risk
assessment of the biohazards done by the principal investigator and is an important
document for laboratory personnel.
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X. Exposure Control Plan (Bloodborne Pathogens and Human
Material)
There are additional requirements for personnel and environmental protection from
bloodborne pathogens (BBP) that may be present in contaminated human blood and
other human materials including human cell lines. This information also applies to nonhuman primate material.
Mississippi State University is committed to providing a safe and healthy work
environment for all staff. In pursuit of this goal, the following Exposure Control Plan
(ECP) is provided to minimize or eliminate occupational exposure to bloodborne
pathogens in accordance with OSHA standard 29 CFR Part 1910.1030, "Occupational
Exposure to Bloodborne Pathogens”.
Most of the controls used to mitigate or eliminate the risk of exposure to BBP will be found
in the general biosafety manual. However, there are some topics specific to BBP that are
covered in the ECP. This ECP includes the following:
•
Program administration
•
Determination of employee exposure
•
Universal precautions
•
Hepatitis B vaccination
•
Post-exposure evaluation and follow-up
•
Procedures for evaluating circumstances surrounding exposure incidents
Program Administration: Please identify the person or persons who will be
responsible for the following: NAME:_______________________
-
Updating the ECP annually;
-
Providing PPE and maintaining safety equipment such as
sharps containers;
-
Training;
-
Liaising with the Longest Student Health Center or other
medical professional;
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-
Managing exposure incidents.
Employee Exposure Determination: The following is a list of job classifications at our
laboratory in which employees have occupational exposure.
Please list all job titles and provide a brief description of their tasks/procedures. Examples given in blue.
Job Title
Research tech
Department/Location
Biology
Lab tech
Biology
Housekeeper
Environmental Services
Task/Procedure
Isolates WBCs
from
blood
Prepares
DNA
from
WBCs
Handles
biohazardous
waste
Add additional lines if needed.
Note: Part-time, temporary, contract and per diem employees are covered by the BBP standard. The
ECP should describe how the standard will be met for these employees.
Universal Precautions: is a work practice control that arose from hospital infection
controls programs. It is designed to prevent the transmission of bloodborne pathogens
such as HIV and HBV when working with human materials. According to the concept of
universal precautions, all human blood and certain human body fluids including human
cell lines are treated as if known to be infectious for HBV, HIV and other bloodborne
pathogens.
Hepatitis B Vaccination
MSU makes available the Hepatitis B Vaccine and vaccination series to all employees
who have occupational exposure, and post-exposure evaluation and follow-up to all
employees who have had an exposure incident.
All medical evaluations and procedures including the hepatitis B vaccine and vaccination
series and post-exposure evaluation and follow-up, including prophylaxis, are made
available at no cost to the employee.
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Hepatitis B Vaccination
Hepatitis B vaccination is made available to the employee after his or her participation in
bloodborne pathogen training.
The vaccine is made available to all employees with occupational exposure unless the
employee has previously received the complete hepatitis B vaccination series, antibody
testing has revealed that the employee is immune, or the vaccine is contraindicated for
medical reasons, or the individual declines. The vaccine can be provided by the Longest
Student Health Center (LSHC). It is recommended that the series begin within 10 days
of initial assignment to all employees identified in the exposure determination section of
this ECP.
All employees who decline to accept hepatitis B vaccination offered by MSU will be
required to sign a Hepatitis B Vaccine Declination form. The laboratory supervisor is
responsible for providing and maintaining the declination form. See Appendix B for an
example. If an employee decides to accept the vaccination at a later date, MSU will make
available hepatitis B vaccination at that time.
To receive the hepatitis B vaccine and vaccination series, contact the Longest Student
Health Center at 325-7539. Information about hepatitis B can be obtained at:
http://www.cdc.gov/ncidod/diseases/hepatitis/b/fact.htm
Post-exposure Evaluation and Follow-up
The Longest Student Health Center will initiate a confidential medical evaluation and
follow-up to an employee following a report of an exposure incident. Employees with an
exposure incident should report to the LSHC, 325-2431.
After an occupational exposure, the following activities will be performed:

Document the routes of exposure and how the exposure occurred

Identify and document the source individual unless the employer can establish that
the identification is not feasible or prohibited by state or local law.
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
Obtain consent and make arrangements to have the source individual tested as
soon as possible to determine HIV, HCV and HBV infectivity; document that the
source individual’s test results were conveyed to the employee’s health care
provider.

If the source individual is already know to be HIV, HCV or HBV positive, new testing
need not be performed.

Assure that the exposed employee is provided with the source individual’s test
results and with information about applicable disclosure laws and regulations
concerning the identity and infectious state of the source individual.

After obtaining consent, collect exposed employee’s blood as soon as possible
after the exposure incident and test blood for HBV and HIV serologic status.

If the employee does not give consent for HIV serological testing during collection
of blood for baseline testing, preserve the baseline blood sample for at least 90
days; if the exposed employee elects to have the baseline sample tested during
this waiting period, perform testing as soon as possible.
Administration of Post-Exposure Evaluation and Follow-Up
Post-exposure follow-up and evaluation requires that the health care professional
evaluating an employee after an exposure receives the following information:

A description of the employee’s job duties relevant to the exposure

Route(s) of exposure

Circumstances of exposure

If possible, the results of the source individual’s blood test

Relevant employee medical records including vaccination history
Procedures for Evaluating the Circumstances Surrounding the Incident
It is necessary to review the exposure incident in or order to prevent or mitigate
subsequent exposures. The following should be reviewed and documented:

Engineering controls in use at the time

Work practices followed

A description of the device being used (if applicable)
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
PPE in use at the time

Location of the incident

Procedure being performed at time of incident

Employee’s training

Record incident in the Sharps Injury Log (See Appendix C for an example).
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Signature and Acknowledgement of Risk
We, the undersigned, understand that the agents used in this facility are potentially
hazardous. We have read and understand this manual and agree to follow the stated
policies and procedures.
Name
Signature
Date
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Appendix A: Example of a Pathogen Safety Data Sheet
STAPHYLOCOCCUS AUREUS
PATHOGEN SAFETY DATA SHEET - INFECTIOUS SUBSTANCES
SECTION I - INFECTIOUS AGENT
NAME: Staphylococcus aureus
SYNONYM OR CROSS REFERENCE: MRSA (methicillin-resistant Staphylococcus aureus),
MSSA (methicillin-susceptive (or sensitive) Staphylococcus aureus), VISA (vancomycinintermediate Staphylococcus aureus), hVISA (heteroresistant vancomycin-intermediate
Staphylococcus aureus), VRSA (vancomycin-resistant Staphylococcus aureus), staph infection,
staphylococcus infection, impetigo, toxic shock syndrome.
CHARACTERISTICS: Staphylococcus aureus are Gram-positive, catalase positive cocci
belonging to the Staphylococcaceae family. They are approximately 0.5-1.5 µm in diameter,
nonmotile, non-spore-forming, facultative anaerobes (with the exception of S. aureus
anaerobius) that usually form in clusters. Many strains produce staphylococcal enterotoxins, the
superantigen toxic shock syndrome toxin (TSST-1), and exfoliative toxins. Staphylococcus
aureus are part of human flora, and are primarily found in the nose and skin.
SECTION II - HAZARD IDENTIFICATION
PATHOGENICITY/TOXICITY: Staphylococcus aureus is an opportunistic pathogen that can
cause a variety of self-limiting to life-threatening diseases in humans. The bacteria are a leading
cause of food poisoning, resulting from the consumption of food contaminated with enterotoxins.
Staphylococcal food intoxication involves rapid onset of nausea, vomiting, abdominal pain,
cramps, and diarrhea. Symptoms usually resolve after 24 hours. Animal bites can result in local
infections, cellulitis, erythema, tenderness, mild fever, adenopathy, and lymphangitis (rarely).
Scalded skin syndrome is caused by exfoliative toxins secreted on the epidermis and mostly
affects neonates and young children. Other skin conditions caused by Staphylococcal exfoliative
toxins include blisters, skin loss, pimples, furuncles, impetigo, folliculitis, abscesses, poor
temperature control, fluid loss, and secondary infection. S. aureus can also cause necrotizing
fasciitis in immunocompromised individuals, although this is very rare. Necrotizing fasciitis is
life-threatening and causes severe morbidity.
Certain strains of S. aureus produce the superantigen TSST-1, which is responsible for 75% of
toxic shock syndrome (TSS) cases. The clinical presentation of TSS is severe and acute
symptoms include high fever, vascular collapse, vomiting, diarrhea, myalgia, hypotension,
erythematous rash, desquamation, and involvement of at least 3 organs. Mortality is very high
and death can occur within 2 hours. Toxic shock syndrome is associated with vaginal
colonization with toxin-producing S. aureus during menstruation, complications with
staphylococcal infection at other sites, or complications of surgical procedures. Deep infections
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include endocarditis, peritonitis, necrotizing pneumonia, bacteremia, meningitis, osteomyelitis,
septic arthritis, and infections of bones, joints and organs.
EPIDEMIOLOGY: Worldwide distribution. Staphylococcus aureus is one of the most common
causes of skin, soft-tissue, and nosocomial infection. Rates of infection in community settings
are increasing. Residents of nursing homes are also at an increased risk of acquiring MRSA.
Around 20% of individuals are persistent carriers of Staphylococcus aureus, about 60% are
intermittent carriers, and approximately 20% rarely carry it. Children are more likely to be
persistent carriers of the bacteria. Young women are at a higher risk for toxic shock syndrome.
HOST RANGE: Humans, wild and domestic animals, including cows.
INFECTIOUS DOSE: At least 100,000 organisms in humans.
MODE OF TRANSMISSION: Ingestion of food containing enterotoxins. Vertical transmission
during vaginal delivery is uncommon. Person-to-person transmission occurs through contact
with a purulent lesion or with a carrier. Unsanitary conditions and crowded community settings
increase exposure to S. aureus. Infection may be spread from person-to-person through health
care workers or patients. Nasal colonization can lead to auto-infection.
INCUBATION PERIOD: Onset of symptoms after consuming contaminated food is usually 30
minutes to 8 hours. Colonies of S. aureus can be carried for an undetermined amount of time;
some individuals may carry it chronically, and some may carry it intermittently.
COMMUNICABILITY: Communicable period is as long as a purulent lesion is present or
carrier state persists.
SECTION III - DISSEMINATION
RESERVOIR: Staphylococcus aureus is found in humans in the nose, groin, axillae, perineal
area (males), mucous membranes, the mouth, mammary glands, hair, and the intestinal,
genitourinary and upper respiratory tracts. Many animals act as reservoirs, particularly cows with
infected udders.
ZOONOSIS: Yes, through direct or indirect contact with an infected animal.
VECTORS: None.
SECTION IV - STABILITY AND VIABILITY
DRUG SUSCEPTIBILITY: Antibiotics such as cloxacillin and cephalexin are commonly used
to treat staph infections. Vancomycin which is administered intravenously is used to treat
MRSA.
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DRUG RESISTANCE: Many strains of Staphylococcus aureus have increasing resistance to
multiple antibiotic classes. Methicillin resistant strains are common causes of nosocomial
infection. Increasing resistance to vancomycin is being documented in many hospitals.
SUSCEPTIBILITY TO DISINFECTANTS: Susceptible to 70% ethanol, clorhexidine, 1%
sodium hypochlorite, 2% glutaraldehyde, 0.25% benzalkonium chloride, and formaldehyde.
PHYSICAL INACTIVATION: Staphylococcus aureus can grow in a pH of 4.2 to 9.3 and in
salt concentrations of up to 15%. Enterotoxins are resistant to temperatures that would destroy
the bacilli. Sensitive to dry heat treatment of 160-170oC for at least an hour, but not to moist heat
treatment.
SURVIVAL OUTSIDE HOST: Survives on carcasses and organs (up to 42 days), floors (less
than 7 days), glass (46 hours), sunlight (17 hours), UV (7 hours), meat products (60 days), coins
(up to 7 days), skin (30 minutes to 38 days) (citation needed). Depending on colony size, S.
aureus can survive on fabrics from days to months.
SECTION V – FIRST AID / MEDICAL
SURVEILLANCE: Monitor for symptoms. In outbreak settings, food poisoning can be
diagnosed on clinical grounds with food cultured for S. aureus. Toxic shock syndrome can be
indicated with a clinical diagnosis and isolation of S. aureus strain, TSST-1, or enterotoxins B or
C. This can be achieved using ELISA, reverse passive latex agglutination, or PCR. Scalded skin
syndrome can be diagnosed clinically, with presence of Nikolsky’s sign and identification of S.
aureus retrieved from the infection site. Bacteremia and deep site infections are confirmed with
direct microscopic examination of clinical specimen.
Note: All diagnostic methods are not necessarily available in all countries.
FIRST AID/TREATMENT: Treatment of abscesses usually does not need antibiotic therapy;
appropriate drainage is usually sufficient. Proper antibiotic therapy is required for more serious
infections.
IMMUNIZATION: None.
PROPHYLAXIS: Elimination of nasal carriage by using topical mupirocin also eliminates hand
carriage.
SECTION VI - LABORATORY HAZARDS
LABORATORY-ACQUIRED INFECTIONS: 29 reported cases as of 1973, with 1 death.
SOURCE/SPECIMENS: Infective stages may be present in CSF, joint aspirates, blood,
abscesses, aerosols, feces, and urine.
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PRIMARY HAZARDS: Trauma of cutaneous barrier, parenteral inoculation, direct
implantation of medical devices (i.e. indwelling catheters and IVs), ingestion of infected
material, and contact with aerosols.
SPECIAL HAZARDS: Contaminated request forms that have been wrapped around specimen
containers. Direct contact with open cuts and lesions of skin.
SECTION VII – EXPOSURE CONTROLS / PERSONAL PROTECTION
RISK GROUP CLASSIFICATION: Risk Group 2.
CONTAINMENT REQUIREMENTS: Containment Level 2 facilities, equipment, and
operational practices for work involving infectious or potentially infectious materials, animals, or
cultures.
PROTECTIVE CLOTHING: Lab coat. Gloves when direct skin contact with infected
materials or animals is unavoidable. Eye protection must be used where there is a known or
potential risk of exposure to splashes.
OTHER PRECAUTIONS: All procedures that may produce aerosols, or involve high
concentrations or large volumes should be conducted in a biological safety cabinet (BSC). The
use of needles, syringes, and other sharp objects should be strictly limited. Additional
precautions should be considered with work involving animals or large scale activities.
SECTION VIII – HANDLING AND STORAGE
SPILLS: Allow aerosols to settle and, wearing protective clothing, gently cover spill with paper
towels and apply an appropriate disinfectant, starting at the perimeter and working towards the
centre. Allow sufficient contact time before clean up.
DISPOSAL: Decontaminate all wastes that contain or have come in contact with the infectious
organism before disposing by autoclave, chemical disinfection, gamma irradiation, or
incineration.
STORAGE: The infectious agent should be stored in leak-proof containers that are appropriately
labelled.
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Appendix B: Example of Hepatitis B Vaccine Declination
I understand that due to my occupational exposure to blood or other potentially infectious
materials I may be at risk of acquiring hepatitis B virus (HBV) infection. I have been given
the opportunity to be vaccinated with hepatitis B vaccine, at no charge to myself.
However, I decline hepatitis B vaccine at this time. I understand that by declining this
vaccine, I continue to be at risk of acquiring hepatitis B, a serious disease. If in the future
I continue to have occupational exposure to blood or other potentially infectious materials,
and I want to be vaccinated with hepatitis B vaccine, I can receive the vaccination series
at no charge to me.
Signed: ___________________________
Date: ________________
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Appendix C: Example of a Sharps Injury Log
The following information, if known, is documented within 14 working days of the date on which each
exposure incident was reported.









Date and time of the exposure incident:___________________________
Date of exposure incident report:________________________________
Report written by:____________________________________________
Type of sharp involved:________________________________________
Description of exposure incident:
•
•
•
•
•




Job classification of exposed employee:_____________________
Department/room where incident occurred:___________________
Procedure being performed by the exposed employee at the time of the incident:
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
_______________________________________________________________________
___________________________________________________
How
did
the
incident
occur:
_______________________________________________________________________
_______________________________________________________________________
______________________________________________________________________
Body part involved:______________________________________
Does the exposed employee believe that any controls (engineering, work practice or administrative)
could have prevented the injury?
_____________________________________________________________________________
_____________________________________________________________________________
___________________________________________________________
Comments on the exposure incident (e.g. additional relevant factors):
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
_________________________________________
Written by:_____________________________ Date:__________________________________
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