· Web viewBIOSAFETY MANUAL. PI. Name. Table of Contents. Intro. duction. to Biosafety. Hazard Identification/Mitigation. Labels and Signs. Decontamination. Waste Disposal

Research Compliance/EHS | 2015

BIOSAFETY MANUALPI Name

Table of ContentsI. Introduction to BiosafetyII. Hazard Identification/MitigationIII. Labels and SignsIV. DecontaminationV. Waste DisposalVI. Transport/ShippingVII. Emergency ResponseVIII. Lab MapIX. IBC ApplicationX. Exposure Control Plan (Bloodborne Pathogens: Human Materials)

AppendixA. Example of a Pathogen Safety Data SheetB. Example of Hepatitis B Vaccine Declination FormC. Example of a Sharps Injury LogD. Animal Biosafety Level 2 Practices

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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 (ECP) section;

3. If your research involves the use of animals, include

Appendix D.

4. Place the manual in a binder and make readily accessible

for all lab personnel;

5. Have all lab personnel read and sign using the signatory

form at the end of the document.

6. Delete any sections that are not related to the research

(e.g. the ECP if not using human materials; ABSL-2

practices if not using animals, etc.)

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I. IntroductionThe 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 TransmissionThere 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 ControlBACKGROUND: 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 CONTROLIdentify 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]6

http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php


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 SignsA 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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http://www.compliancesigns.com/OWE-1461.shtml

http://www.labsafety.com/search/biohazard+signs/29947/





IV. DecontaminationThis 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-SOP-

001 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.


If the risk assessment requires the use of reusable PPE (safety goggles, face shields) describe

the procedure for cleaning and decontamination.


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V. Waste DisposalAll 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


biohazardous waste and must be disposed of in accordance with the Guidelines. See IBC-

SOP-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.14


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/ShippingTransport 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


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 NumbersPrincipal Investigator: Office:

Cell :

Alternate Contact: Office:Cell :

Emergency 911University Police 662-325-2121Student 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 662-325-3294Radiation 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 SpillA 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 ResponseEach 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 Body1. 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 Cabinet1. 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 BSC1. 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 Centrifuge1. Close lid!

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2. Clear area of personnel.

3. Wait at least 30 minutes for aerosols to settle before clean-up.

4. Wear a lab coat, gloves, and safety glasses during clean-up.

5. If broken glass or other contaminated sharp is present, use forceps to remove,

NOT YOUR HANDS.

6. Wipe rotors and buckets with disinfectant then remove to nearest BSC for more

extensive decontamination.

7. Use paper towels to absorb then thoroughly soak inside of the centrifuge with an

appropriate disinfectant using the manufacturer’s recommended minimum

contact time.

8. Dispose of contaminated materials using appropriate biohazardous waste

disposal procedures.

Outside the Laboratory, In Transit1. 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.

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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

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 Spill1. 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 Spill1. 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.

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F. Medical Emergency1. If an individual is injured, call 911. Employees are not required to perform first aid.

2. Notify supervisor

3. Do not move person or perform any first aid if you are not trained and qualified.

G. Animal Bites1. Clean bite thoroughly with soap and water.

2. Apply bandage

3. Notify supervisor

4. Obtain medical attention.

H. Tornado or Hurricane1. 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 emergenciesIn 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 concerns1. Do not attempt to repair failure (e.g. burst pipe, elevator stuck, temperature, power

outage, etc.,)

2. Call 325-2005

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3. Give name, location, and state problem. Stay on phone until information

acknowledged.

K. Suspicious Persons Or ActivitiesAny suspicious persons or activities must be immediately reported to MSU PD at 325-

2121. Do not open the laboratory door to allow any suspicious or unknown persons

entry to the laboratory.

L . Oral Threat1. 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.

25


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.

26


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 non-

human 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 Department/Location Task/ProcedureResearch associate Biology Isolates WBCs from

bloodPost-Doc Biology Prepares DNA from

WBCsUndergrad student Basic Sciences Handles biohazardous

wasteAdd 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 VaccinationHepatitis 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-upThe 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-UpPost-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 IncidentIt 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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Exposure Control Plan Annual Review

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

Date: Initials:

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Signature and Acknowledgement of Risk

We, the undersigned, understand that the biologic materials 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 (vancomycin-intermediate 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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Appendix D

BMBL 5th Edition Section V: Vertebrate Animal Biosafety Level Criteria for Vivarium Research Facilities

This guidance is provided for the use of experimentally infected animals housed in indoor research facilities (e.g., vivaria), and is also useful in the maintenance of laboratory animals that may naturally harbor zoonotic infectious agents. In both instances, the institutional management must provide facilities, staff, and established practices that reasonably ensure appropriate levels of environmental quality, safety, security and care for the laboratory animal. Laboratory animal facilities are a special type of laboratory. As a general principle, the biosafety level (facilities, practices, and operational requirements) recommended for working with infectious agents in vivo and in vitro are comparable.

The animal room can present unique problems. In the animal room, the activities of the animals themselves can present unique hazards not found in standard microbiological laboratories. Animals may generate aerosols, they may bite and scratch, and they may be infected with a zoonotic agent. The co-application of Biosafety Levels and the Animal Biosafety Levels are determined by a protocol-driven risk assessment. These recommendations presuppose that laboratory animal facilities, operational practices, and quality of animal care meet applicable standards and regulations (e.g., Guide for the Care and Use of Laboratory Animals and Laboratory Animal Welfare Regulations) and that appropriate species have been selected for animal experiments. In addition, the organization must have an occupational health and safety program that addresses potential hazards associated with the conduct of laboratory animal research. The following publication by the Institute for Laboratory Animal Research (ILAR), Occupational Health and Safety in the Care and Use of Research Animals, is most helpful in this regard. Additional safety guidance on working with non-human primates is available in the ILAR publication, Occupational Health and Safety in the Care and Use of Nonhuman Primates.

Facilities for laboratory animals used in studies of infectious or non-infectious disease should be physically separate from other activities such as animal production and quarantine, clinical laboratories, and especially from facilities providing patient care. Traffic flow that will minimize the risk of cross contamination should be incorporated into the facility design.

Animal Biosafety Level 2Animal Biosafety Level 2 builds upon the practices, procedures, containment, equipment, and facility requirements of ABSL-1. ABSL-2 is suitable for work involving laboratory animals infected with agents associated with human disease and pose moderate hazards to personnel and the environment. It also addresses hazards from ingestion as well as from percutaneous and mucous membrane exposure.

ABSL-2 requires that: 1) access to the animal facility is restricted; 2) personnel must have specific training in animal facility procedures, the handling of infected animals and the manipulation of pathogenic agents;

40


3) personnel must be supervised by individuals with adequate knowledge of potential hazards, microbiological agents, animal manipulations and husbandry procedures; and 4) BSCs or other physical containment equipment is used when procedures involve the manipulation of infectious materials, or where aerosols or splashes may be created.

Appropriate personal protective equipment must be utilized to reduce exposure to infectious agents, animals, and contaminated equipment. Implementation of employee occupational health programs should be considered.

The following standard and special practices, safety equipment, and facility requirements apply to ABSL-2.

A.Standard Microbiological Practices1. The animal facility director establishes and enforces policies, procedures, and

protocols for institutional policies and emergencies. Each organization must assure that worker safety and health concerns are addressed as part of the animal protocol review. Prior to beginning a study, animal protocols must also be reviewed and approved by the IACUC and the Institutional Biosafety Committee.

2. A safety manual specific to the animal facility is prepared or adopted in consultation with the animal facility director and appropriate safety professionals. The safety manual must be available and accessible. Personnel are advised of potential hazards, and are required to read and follow instructions on practices and procedures. Consideration should be given to specific biohazards unique to the animal species and protocol in use.

3. The supervisor must ensure that animal care, laboratory, and support personnel receive appropriate training regarding their duties, animal husbandry procedure, potential hazards, manipulations of infectious agents, necessary precautions to prevent hazard or exposures, and hazard/exposure evaluation procedures (physical hazards, splashes, aerosolization, etc.). Personnel must receive annual updates or additional training when procedures or policies change. Records are maintained for all hazard evaluations, employee training sessions and staff attendance.

4. An appropriate medical surveillance program is in place, as determined by risk assessment. The need for an animal allergy prevention program should be considered. Facility supervisors should ensure that medical staff is informed of potential occupational hazards within the animal facility, to include those associated with research, animal husbandry duties, animal care and manipulations. Personal health status may impact an individual’s susceptibility to infection, ability to receive immunizations or prophylactic interventions. Therefore, all personnel and particularly women of childbearing age should be provided 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. Personnel using respirators must be enrolled in an appropriately constituted respiratory protection program.

5. A sign incorporating the universal biohazard symbol must be posted at the entrance to areas where infectious materials and/ or animals are housed or are manipulated when infectious agents are present. The sign must include the animal biosafety level, general occupational health requirements, personal protective equipment

41


requirements, the supervisor’s name, telephone number and required procedures for entering and exiting the animal areas.Security-sensitive agent information and occupational health requirements should be posted in accordance with the institutional policy.Advance consideration should be given to emergency and disaster recovery plans, as a contingency for man-made or natural disasters.

6. Access to the animal room is limited. Only those persons required for program or support purposes are authorized to enter the animal facility and the areas where infectious materials and/or animals are housed or manipulated. All persons including facility personnel, service workers, and visitors are advised of the potential hazards (physical, naturally occurring, or research pathogens, allergens, etc.) and are instructed on the appropriate safeguards.

7. Protective laboratory coats, gowns, or uniforms are recommended to prevent contamination of personal clothing. Gloves are worn to prevent skin contact with contaminated, infectious and hazardous materials and when handling animals. Gloves and personal protective equipment should be removed in a manner that prevents transfer of infectious materials outside of the areas where infectious materials and/or animals are housed or are manipulated. Persons must wash their hands after removing gloves, and before leaving the areas where infectious materials and/or animals are housed or are manipulated. Eye, face and respiratory protection should be used in rooms containing infected animals, as dictated by the risk assessment.

8. 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 of the laboratory in cabinets or refrigerators designated and used for this purpose.

9. All procedures are carefully performed to minimize the creation of aerosols or splatters of infectious materials and waste.

10. Mouth pipetting is prohibited. Mechanical pipetting devices must be used.11. Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and

broken glassware must be developed and implemented. When applicable, laboratory supervisors should adopt improved engineering and work practice controls that reduce the risk of sharps injuries. Precautions must always be taken with sharp items. These include: The use of needles and syringes or other sharp instruments in the animal facility is limited to situations where there is no alternative such as parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. Disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal. Used, disposable needles must be carefully placed in puncture-resistant containers used for sharps disposal. Sharps containers should be located as close to the work site as possible. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving.Broken glassware must not be handled directly; it should be removed using a brush and dustpan, tongs, or forceps. Plastic ware should be substituted for glassware whenever possible. Use of equipment with sharp edges and corners should be avoided.

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12. Equipment and work surfaces are routinely decontaminated with an appropriate disinfectant after work with an infectious agent, and after any spills, splashes, or other overt contamination.

13. Animals and plants not associated with the work being performed must not be permitted in the areas where infectious materials and/ or animals are housed or manipulated.

14. An effective integrated pest management program is required. 15. All wastes from the animal room (including animal tissues, carcasses, and bedding)

are transported from the animal room in leak-proof containers for appropriate disposal in compliance with applicable institutional, local and state requirements.Decontaminate all potentially infectious materials before disposal.

B. Special Practices1. Animal care staff, laboratory and routine support personnel must be provided a

medical surveillance program as dictated by the risk assessment and administered appropriate immunizations for agents handled or potentially present, before entry into animal rooms. When appropriate, a base line serum sample should be stored.

2. Procedures involving a high potential for generating aerosols should be conducted within a biosafety cabinet or other physical containment device. When a procedure cannot be performed within a biosafety cabinet, a combination of personal protective equipment and other containment devices must be used. Restraint devices and practices that reduce the risk of exposure during animal manipulations (e.g., physical restraint devices, chemical restraint medications) should be used whenever possible.

3. Decontamination by an appropriate method (e.g. autoclave, chemical disinfection, or other approved decontamination methods) is necessary for all potentially infectious materials and animal waste before movement outside the areas where infectious materials and/or animals are housed or are manipulated. This includes potentially infectious animal tissues, carcasses, contaminated bedding, unused feed, sharps, and other refuse. A method for decontaminating routine husbandry equipment, sensitive electronic and medical equipment should be identified and implemented. Materials to be decontaminated outside of the immediate areas where infectious materials and/or animals are housed or are manipulated must be placed in a durable, leak proof, covered container and secured for transport. The outer surface of the container is disinfected prior to moving materials. The transport container must have a universal biohazard label. Develop and implement an appropriate waste disposal program in compliance with applicable institutional, local and state requirements. Autoclaving of content prior to incineration is recommended.

4. Equipment, cages, and racks should be be handled in a manner that minimizes contamination of other areas. Equipment must be decontaminated before repair, maintenance, or removal from the areas where infectious materials and/or animals are housed or are manipulated.

5. Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material.

6. Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the safety manual. All such incidents must be reported to the animal facility supervisor or personnel

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designated by the institution. Medical evaluation, surveillance, and treatment should be provided as appropriate and records maintained.

C. Safety Equipment (Primary Barriers and Personal Protective Equipment)1. Properly maintained BSCs, personal protective equipment (e.g., gloves, lab coats,

face shields, respirators, etc.) and/or other physical containment devices or equipment, are used whenever conducting procedures with a potential for creating aerosols, splashes, or other potential exposures to hazardous materials. These include necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, and intranasal inoculation of animals. When indicated by risk assessment, animals are housed in primary biosafety containment equipment appropriate for the animal species, such as solid wall and bottom cages covered with filter bonnets for rodents or other equivalent primary containment systems for larger animal cages.

2. A risk assessment should determine the appropriate type of personal protective equipment to be utilized. Scrub suits and uniforms are removed before leaving the animal facility. Reusable clothing is appropriately contained and decontaminated before being laundered. Laboratory and protective clothing should never be taken home. Gowns, uniforms, laboratory coats and personal protective equipment are worn while in the areas where infectious materials and/or animals are housed or manipulated and removed prior to exiting. Disposable personal protective equipment and other contaminated waste are appropriately contained and decontaminated prior to disposal. Eye and face protection (mask, goggles, face shield or other splatter guard) are used for manipulations or activities that may result in splashes or sprays from infectious or other hazardous materials and when the animal or microorganisms must be handled outside the BSC or containment device. Eye and face protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse. Persons who wear contact lenses should also wear eye protection when entering areas with potentially high concentrations or airborne particulates. Persons having contact with NHPs should assess risk of mucous membrane exposure and wear protective equipment (e.g., masks, goggles, face shields) appropriate for the task to be performed. Respiratory protection is worn based upon risk assessment.

3. Gloves are worn to protect hands from exposure to hazardous materials. A risk assessment should be performed to identify the appropriate glove for the task and alternatives to latex gloves should be available. Gloves are changed when contaminated, glove integrity is compromised, or when otherwise necessary. Gloves must not be worn outside the animal rooms. Gloves and personal protective equipment should be removed in a manner that prevents transfer of infectious materials. Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated waste. Persons must wash their hands after handling animals and before leaving the areas where infectious materials and/or animals are housed or are manipulated. Hand washing should occur after the removal of gloves.

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D. Laboratory Facilities (Secondary Barriers)1.The animal facility is separated from areas that are open to unrestricted personnel traffic within the building. External facility doors are self-closing and self-locking. Doors to areas where infectious materials and/or animals are housed, open inward, are self-closing, are kept closed when experimental animals are present, and should never be propped open. Doors to cubicles inside an animal room may open outward or slide horizontally or vertically.2. A hand-washing sink is located at the exit of the areas where infectious materials and/or animals are housed or are manipulated. Additional sinks for hand washing should be located in other appropriate locations within the facility. If the animal facility has segregated areas where infectious materials and/or animals are housed or manipulated, a sink must also be available for hand washing at the exit from each segregated area. Sink traps are filled with water, and/or appropriate disinfectant to prevent the migration of vermin and gases.3. The animal facility is designed, constructed, and maintained to facilitate cleaning and housekeeping. The interior surfaces (walls, floors and ceilings) are water resistant.Penetrations in floors, walls and ceiling surfaces are sealed, including openings around ducts, doors and doorframes, to facilitate pest control and proper cleaning. Floors must be slip-resistant, impervious to liquids, and resistant to chemicals. Cabinets and bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals. Spaces between benches, cabinets, and equipment should be accessible for cleaning. Furniture should be minimized. Chairs used in animal area must be covered with a non-porous material that can be easily cleaned and decontaminated. Furniture must be capable of supporting anticipated loads and uses. Sharp edges and corners should be avoided.4. External windows are not recommended; if present, windows must be sealed and resistant to breakage. The presence of windows may impact facility security and therefore should be assessed by security personnel.5.Ventilation should be provided in accordance with the Guide for Care and Use of Laboratory Animals. The direction of airflow into the animal facility is inward; animal rooms maintain inward directional airflow compared to adjoining hallways. A ducted exhaust air ventilation system is provided. Exhaust air is discharged to the outside without being recirculated to other rooms. Ventilation system design should consider the heat and high moisture load produced during the cleaning of animal rooms and the cage wash process.6.Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, are arranged to minimize horizontal surface areas, to facilitate cleaning and minimize the accumulation of debris or fomites.7. Floor drains must be maintained and filled with water, and/or appropriate disinfectant to prevent the migration of vermin and gases. Cages should be autoclaved or otherwise decontaminated prior to washing. Mechanical cage washer should have a final rinse temperature of at least 180°F. The cage wash area should be designed to accommodate the use of high-pressure spray systems, humidity, strong chemical disinfectants and 180°F water temperatures during the cage/equipment cleaning process.8.Illumination is adequate for all activities, avoiding reflections and glare that could impede vision.

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9. If BSCs are present, they 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, heavily traveled laboratory areas, and other possible airflow disruptions. HEPA filtered exhaust air from a Class II BSC can be safely re-circulated 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 to the outside through an independent, hard connection. Provisions to assure proper safety cabinet performance and air system operation must be verified. BSCs should be recertified at least once a year to ensure correct performance. All BSCs should be used according to manufacturer’s specifications to protect the worker and avoid creating a hazardous environment from volatile chemicals and gases.10. If vacuum service (i.e., central or local) is provided, each service connection should be fitted with liquid disinfectant traps and an in-line HEPA filter placed as near as practicable to each use point or service cock. Filters are installed to permit in-place decontamination and replacement.11. An autoclave should be present in the animal facility to facilitate decontamination of infectious materials and waste.12. Emergency eyewash and shower are readily available; location is determined by risk assessment.

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Documents

· Web viewBIOSAFETY MANUAL. PI. Name. Table of Contents. Intro. duction. to Biosafety. Hazard Identification/Mitigation. Labels and Signs. Decontamination. Waste Disposal