2013 Biological Safety Manual

EHS University of Central Oklahoma 1/1/2013

January 1, 2013

BIOLOGICAL SAFETY MANUAL

Table of Contents Table of Contents....................................................................................................................... 2 BIOLOGICAL SAFETY .............................................................................................................. 4 A. Biological Risk Assessment ................................................................................................... 4 B. General Laboratory Practice .................................................................................................. 6 C. Biological Safety Level 1 (BSL-1) .......................................................................................... 7 1. Standard Microbiological Practices (BSL-1) ........................................................................ 7 2. Special Practices: None (BLS-1) ........................................................................................ 9 3. Safety Equipment (Primary Barriers) (BLS-1) ..................................................................... 9 4. Laboratory Facilities (Secondary Barriers) (BLS-1) ............................................................. 9 D. Biological Safety Level 2 (BSL-2) .........................................................................................10 1.

Standard Microbiological Practices (BSL-2)....................................................................10

2. Special Practices (BSL-2) ..................................................................................................12 3. Safety Equipment (Primary Barriers) (BSL-2) ....................................................................12 4. Laboratory Facilities (Secondary Barriers) (BSL-2) ............................................................13 E. Biological Safety Level 3 (BSL-3)..........................................................................................15 1. Standard Microbiological Practices (BSL-3) .......................................................................15 2. Special Practices (BSL-3) ..................................................................................................17 3. Safety Equipment (Primary Barriers) (BSL-3) ....................................................................18 4. Laboratory Facilities (Secondary Barriers) (BSL-3) ............................................................19 F. Biological Safety Level 4 .......................................................................................................21 G. Animal Biosafety Level 1 (ABSL-1) .......................................................................................21 1. Standard Practices (ABSL-1) .............................................................................................21 2. Special Practices: None required. (ABSL-1) ......................................................................23 3. Safety Equipment (Primary Barriers and Personal Protective Equipment) (ABSL-1)..........23 4. Animal Facilities (Secondary Barriers) (ABSL-1)................................................................24 H. Animal Biological Safety Level 2 (ABSL-2) ...........................................................................25 1. Standard Practices (ABSL-2) .............................................................................................25 2. Special Practices (ABSL-2) ...............................................................................................27 3. Safety Equipment (Primary Barriers and Personal Protective Equipment) .........................28 4. Animal Facilities (Secondary Barriers) (ABSL-2) ................................................................30 I. Animal Biological Safety Level 3 (ABSL-3) .............................................................................32 1. Standard Practices (ABSL-3) .............................................................................................32

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2. Special Practices (ABSL-3) ...............................................................................................34 3. Safety Equipment (Primary Barriers and P.P.E.) (ABSL-3) ................................................36 4. Laboratory Facilities (Secondary Barriers) (ABSL-3) .........................................................37 J. Animal Biological Safety Level 4 (ABSL-4) ............................................................................40 K. Guidelines for Work with Toxins of Biological Origin .............................................................40 Sources: ............................................................................................................................43 L.

Procedures for Working with Human Blood or Other Potentially Infectious Material ...........44

M. Recombinant DNA ...............................................................................................................48 Source: NIH Guidelines for Research Involving Recombinant DNA Molecules, April, 2002. ..........................................................................................................................................49 N. Institutional Biological Safety Committee ..............................................................................49 O. Animal Safety .......................................................................................................................51 1. Zoonotic Diseases .............................................................................................................51 2.

Injury ..............................................................................................................................52 Sources: ............................................................................................................................54

TABLE VI-1 ...............................................................................................................................55 DISEASES, PROTECTIVE EQUIPMENT, AND MEDICAL MONITORING/VACCINATIONS FOR CERTAIN ANIMAL SPECIES........................................................................................55 TABLE VI-2 ...............................................................................................................................57 RECOMMENDED SAFE WORK PRACTICES ......................................................................57 TABLE VI-2 ...............................................................................................................................58 RECOMMENDED SAFE WORK PRACTICES ......................................................................58 TABLE VI-3 ...............................................................................................................................59 ADDITIONAL PRECAUTIONS FOR ANIMAL BIOSAFETY LEVELS 2 AND 3 ......................59

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BIOLOGICAL SAFETY Microbiological and biomedical laboratories are special, often unique, work environments that may pose special infectious disease risks to persons in or near them. Personnel have contracted infections in the laboratory throughout the history of microbiological and biomedical research. A number of cases have been attributed to carelessness or poor technique in the handling of infectious materials. The most important element of safety is strict adherence to standard microbiological practices and techniques. The Centers for Disease Control and Prevention (CDC) and the National Institutes of Health (NIH) have established four biosafety levels which consist of combinations of laboratory practices and techniques, safety equipment, and laboratory facilities appropriate for the operations performed and the hazard posed by infectious agents or recombinant DNA research. The Principle Investigator is responsible for making a determination of the required levels of physical and biological containment required for the work to be performed through the risk assessment process. When standard laboratory practices are not sufficient to control the hazard associated with a particular agent or laboratory procedure, additional measures may be needed. Vertebrate animal biosafety level criteria are also provided which describe four combinations (designated Animal Biosafety Levels 1-4) of practices, safety equipment, and facilities for experiments on animals infected with agents which produce, or may produce, human infection. These guidelines are also useful in the maintenance of laboratory animals that may naturally harbor zoonotic infectious diseases. Finally, guidelines for work with toxins of biological origin are presented. Toxins are unique in that they do not replicate, are not infectious, and are difficult to transmit mechanically or manually from person to person. Therefore, toxins are usually handled using established general guidelines for toxic or highly toxic chemicals with the incorporation of additional safety and security measures based upon a risk assessment for each specific laboratory operation. Each laboratory should develop standard operating procedures (SOPs) which identify the hazards that will or may be encountered and which specify practices and procedures designed to minimize or eliminate risks. Guidelines for developing these SOPs are provided here.

A. Biological Risk Assessment Risk assessment is a process used to identify the hazardous characteristics of a known infectious or potentially infectious agent or material, the activities that can result in a person’s exposure to an agent, the likelihood that such exposure will cause a laboratory acquired infection, and the probable consequences of such an infection. The information identified by risk assessment will provide a guide for the selection of appropriate biosafety levels and microbiological practices, safety equipment, and facility safeguards that can prevent laboratory acquired infections. Biological risk assessment is a subjective process requiring consideration of many hazardous characteristics of agents and procedures. Below is an outline of some of the considerations that should be evaluated.

1. Consider the principal hazardous characteristics of the agent, which include its capability to infect and cause disease in a susceptible human host, severity of disease, and the

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availability of preventive measures and effective treatments. Make a preliminary determination of the biosafety level that best correlates with the initial risk assessment based on the identification and evaluation of the agent hazards. Remember that aerosol and droplet routes of agent transmission also are important considerations in specification of safety equipment and facility design that result in a given biosafety level. 2. Identify laboratory procedure hazards. The principal laboratory procedure hazards are agent concentration, suspension volume, equipment and procedures that generate small particle aerosols and larger airborne particles (droplets), and use of sharps. Procedures involving animals can present a number of hazards such as bites and scratches, exposure to zoonotic agents, and the handling of experimentally generated infectious aerosols. The complexity of a laboratory procedure can also present an increased hazard. 3. Make a final determination of the appropriate biosafety level and select additional precautions indicated by the risk assessment. 4. The final selection of the appropriate biosafety level and the selection of any additional laboratory precautions require a comprehensive understanding of the practices, safety equipment, and facility safeguards described the following sections of this manual. There will be situations where the intended use of an agent requires greater precautions. It is also important to recognize that individuals in the laboratory may differ in their susceptibility to disease. Preexisting diseases, medications, compromised immunity, and pregnancy or breast-feeding that may increase exposure to infants to certain agents, are some of the conditions that may increase the risk of an individual for acquiring a laboratory acquired infection. Consultation with an occupational physician knowledgeable in infectious diseases is advisable in these circumstances. 5. Evaluate the proficiencies of staff regarding safe practices and the integrity of safety equipment. The protection of laboratory workers, other persons associated with the laboratory, and the public will depend ultimately on the laboratory workers themselves. In conducting a risk assessment, the laboratory director or principal investigator should ensure that laboratory workers have acquired the technical proficiency in the use of microbiological practices and safety equipment required for the safe handling of the agent, and have developed good habits that sustain excellence in the performance of those practices. An evaluation of a person’s training, experience in handling infectious agents, proficiency in the use of sterile techniques and biological safety cabinets (BSCs), ability to respond to emergencies, and willingness to accept responsibility for protecting one’s self and others is important insurance that a laboratory worker is capable of working safely. The laboratory director or principal investigator should also ensure that the necessary safety equipment is available and operating properly. 6. Review the risk assessment with a biosafety professional or subject matter expert, and ultimately, with the UCO Institutional Biosafety Committee (IBC). A review of the risk assessment and selected safeguards by other knowledgeable individuals will promote the use of safe practices in work with hazardous agents in microbiological and biomedical laboratories.

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B. General Laboratory Practice 1. Engineering controls such as biological safety cabinets (BSCs) should be examined and maintained or replaced on a regular schedule to ensure their effectiveness. It is imperative that Class I and II BSCs are tested and certified in place at the time of installation within the laboratory, at any time the cabinet is moved, and at least annually thereafter. 2. Employees should wash their hands immediately or as soon as possible after removal of gloves or other personal protective equipment and after hand contact with blood or other potentially infectious materials. 3. All personal protective equipment should be removed immediately upon leaving the work area or as soon as possible, if overtly contaminated, and placed in an appropriately designated area or container for storage, washing, decontamination, or disposal. 4. Immediately after use, contaminated sharps should be disposed in closable, punctureresistant containers which are leak-proof on the sides and bottom and color-coded or labeled with the biohazard symbol. 5. Sharps containers should be easily accessible to personnel and located in the area of use. 6. Used needles and other sharps should not be sheared, bent, broken, recapped, or resheathed by hand. 7. Eating, drinking, smoking, and applying cosmetics or lip balm, and handling contact lenses are prohibited in areas where work with biohazardous materials is performed. 8. Food and drink must not be stored in refrigerators, freezers, or cabinets where biohazardous materials are stored. 9. All procedures involving biohazardous materials should be performed in such a manner as to minimize splashing, spraying, and aerosolization of these substances. Mouth pipetting/suctioning is prohibited. 10. Broken glassware which may be contaminated should not be picked up directly with the hands. It should be cleaned up using mechanical means such as a brush and dust pan, tongs, cotton swabs or forceps. 11. All infectious waste should be disposed in accordance with the procedures found in Section VIII., "Biomedical Waste”. 12. House vacuum systems should be protected from aspirations of infectious fluids. For Biosafety Level 2 laboratory work, an in-line flask containing a suitable decontamination solution should be used, serving as a fluid overflow collection vessel, connected to the vacuum system. For Biosafety Level 3 laboratory work, two flasks containing a decontamination solution prior to the HEPA filter should be used. See Section II.I. House Vacuum System Protection”.

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13. Biological hazard (biohazard) tags or labels should be used to identify the actual or potential presence of a biological hazard and to identify equipment, containers, rooms, experimental animals, or combinations thereof, those contains or are contaminated with hazardous biological agents. Sources: CDC/NIH Primary Containment for Biohazards: Selection, Installation, and use of Biological Safety Cabinets, U.S. Department of Health and Human Services Public Health Services, Centers for Disease Control and Prevention and National Institutes of Health, September 1995 OSHA Specifications for Accident Prevention Signs and Tags Standard (29 CFR 1910.145)

C. Biological Safety Level 1 (BSL-1) Biosafety Level 1 (BSL-1) is suitable for work involving well-characterized agents not known to cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environment. BSL-1 laboratories are not necessarily separated from the general traffic patterns in the building. Work is typically conducted on open bench tops using standard microbiological practices, but may be used as determined by appropriate risk assessment. Special containment equipment or facility design is not required nor generally used. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or a related science. The following standard and special practices, safety equipment and facilities apply to agents assigned to BSL-

1. Standard Microbiological Practices (BSL-1) a. Access to the laboratory should be limited or restricted at the discretion of the laboratory director when experiments or work with cultures and specimens are in progress. b. The Principal Investigator (PI) must enforce the institutional policies that control access to the laboratory. c. Persons must wash their hands after working with organisms containing recombinant DNA molecules or potentially hazardous materials, after removing gloves, and before leaving the laboratory. d. 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 work area in cabinets or refrigerators designated and used for this purpose only. e. Mouth pipetting is prohibited; mechanical pipetting devices are used. f.

Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken glassware are developed and implemented. Whenever practical, PIs 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:

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BIOLOGICAL SAFETY MANUAL (1) 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. (2) Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal. (3) Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. (4) Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plasticware should be substituted for glassware whenever possible.

g.

All procedures should be performed carefully to minimize the creation of splashes or aerosols.

h. Work surfaces should be decontaminated at least once a day and after any spill of viable or potentially infectious material with an appropriate disinfectant. i.

All cultures, stocks, other potentially infectious materials, and other regulated wastes are decontaminated before disposal by an approved decontamination method, such as autoclaving.

j.

Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak-proof container and closed for transport from the laboratory. Materials to be removed from the facility for decontamination must be packed in accordance with applicable local, state, and federal regulations.

k. A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. The sign may include the name of the agent(s) in use and the name and phone number of the PI or other responsible personnel. Agent information should be posted in accordance with institutional policy, if applicable. l.

An integrated pest management program is in effect.

m. The PI must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. n. 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 child-bearing age should be provided with information regarding immune competence and conditions that may predispose them to infection. o. Individuals having these conditions should be encouraged to self-identify to Employee Health for appropriate counseling and guidance.

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2. Special Practices: None (BLS-1) 3. Safety Equipment (Primary Barriers) (BLS-1) a. Special containment devices or equipment such as a BSC are generally not required for manipulations of agents assigned to BSL-1. b. Laboratory coats, gowns, or uniforms are recommended to be worn to prevent contamination of personal clothing. c. Protective eyewear should be worn when conducting procedures that have the potential to create splashes of microorganisms or other hazardous materials. Persons who wear contact lenses should also wear eye protection. d. Gloves must be worn to protect hands from exposure to hazardous materials. (1) Glove selection should be based on an appropriate risk assessment. (2) Alternatives to latex gloves should be available. (3) Laboratory workers should change gloves when contaminated, integrity has been compromised, or when otherwise necessary. Laboratory workers should remove gloves and wash hands when work with hazardous materials has been completed and before leaving the laboratory (4) Disposable gloves should not be washed or reused. Used gloves should be disposed with other contaminated laboratory waste. (5) Hand washing protocols must be rigorously followed.

4. Laboratory Facilities (Secondary Barriers) (BLS-1) a. Laboratories should have doors for access control. b. Laboratories must have a sink for hand washing. c. The laboratory should be designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate. d. Laboratory furniture must be capable of supporting anticipated loading and uses. Spaces between benches, cabinets, and equipment are accessible for cleaning. e. Bench tops must be impervious to water and are resistant to moderate heat and the organic solvents, acids, alkalis, and other chemicals (such as those used to decontaminate the work surface and equipment). f.

Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with an appropriate disinfectant.

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g. Laboratory windows that open to the exterior should be fitted with screens. End of BSL-1 Section

D. Biological Safety Level 2 (BSL-2) Biosafety Level 2 (BSL-2) is suitable for work involving agents that pose moderate potential hazard to personnel and the environment. It differs from BSL-1 in that (1) laboratory personnel should have specific training in handling pathogenic agents and be supervised by scientists competent in handling infectious agents and associated procedures; (2) access to the laboratory is limited when work is being conducted; and (3) all procedures in which infectious aerosols or splashes may be created should be conducted in BSCs or other physical containment equipment. The following standard and special practices, safety equipment, and facilities apply to BSL-2.

1. Standard Microbiological Practices (BSL-2) THE PI MUST ENFORCE THE INSTITUTIONAL POLICIES THAT CONTROL ACCESS TO THE LABORATORY. a. Persons must wash their hands after working with potentially hazardous materials or animals, after removing gloves, and before leaving the laboratory. b. Eating, drinking, smoking, handling contact lenses, and applying cosmetics are not permitted in the work areas. Storing food for human consumption is not permitted in the laboratory. Food must be stored outside the laboratory area in cabinets or refrigerators designated for this purpose only c. Mouth pipetting is prohibited; mechanical pipetting devices must be used. d. Policies for the safe handling of sharps such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, PIs 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: (1) Hypodermic needles and syringes should only be used for parenteral injection and aspiration of fluids from laboratory animals or diaphragm bottles. Only needle-locking syringes or disposable syringe-needle unites (i.e., needle is integral to the syringe) may be used for the injection or aspiration of fluids containing microorganisms or viruses. (2) Safe needle devices should be used whenever such a device is available for the task requiring the use of a needle/syringe.

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(3) 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. (4) Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal. (5) Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. (6) Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plasticware should be substituted for glassware whenever possible. f.

All procedures should be performed carefully to minimize the creation of splashes or aerosols.

g. Work surfaces should be decontaminated after completion of work or after any spill or splash of potentially infectious material with an appropriate disinfectant that is effective against the agent(s) of concern. h. All cultures, stocks, other potentially infectious materials, and other regulated wastes should be decontaminated before disposal by an approved method, such as autoclaving. (1) Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak-proof container and secured for transport from the facility. (2) Materials to be removed from the facility for decontamination must be packaged in accordance with applicable local, state, and federal regulations. i.

A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present or organisms containing recombinant DNA molecules are in use. 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.

j.

An effective integrated pest management program is required.

k. The PI must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. l.

Personnel must receive annual updates or additional training when procedural or policy changes occur.

m. 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 child-bearing age should be provided with information regarding immune competence and conditions that may predispose them to infection.

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Individuals having these conditions should be encouraged to self-identify to Employee Health for appropriate counseling and guidance.

2. Special Practices (BSL-2) a. All persons entering the laboratory must be advised of the potential hazards and meet specific entry/exit requirements. b. Laboratory personnel must be provided medical surveillance and offered appropriate immunizations for agents handled or potentially present in the laboratory. c. If applicable to the research being performed, policies and procedures describing the collection and storage of serum samples from at-risk personnel should be developed. d. Laboratory-specific biosafety manual/SOPs must be prepared and adopted as laboratory policy. The biosafety manual/SOPs must be available and accessible. e. The PI must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices before working with BSL-2 agents. f.

Potentially infectious materials must be placed in a durable, leak proof container during collection, handling, processing, storage, or transport within a facility.

g. Laboratory equipment should be routinely decontaminated and decontaminated after spills, splashes, or other potential contamination. (1) Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material. (2) Equipment must be decontaminated before repair, maintenance, or removal from the laboratory. h. Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the laboratory biosafety safety manual/SOPs. All such incidents must be reported to the PI and to IBC. Medical evaluation, surveillance, and treatment should be provided through Employee Health or the nearest emergency room, and appropriate records maintained. i.

Animals and plants not involved in the work being performed are not permitted in the lab.

j.

All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted within a BSC or other physical containment devices.

3. Safety Equipment (Primary Barriers) (BSL-2) a. Properly maintained BSCs (preferably Class II), other appropriate personal protective equipment, or other physical containment devices must be used whenever:

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(1) 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 embryonate eggs. (2) 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 are used, and if these rotors or safety cups are opened only in a BSC. b. Protective laboratory coats, gowns, smocks, or uniforms designated for lab use must be worn when working with hazardous materials. This protective clothing should be removed and left in the laboratory before leaving for non-laboratory areas (e.g., cafeteria, library, administrative offices). All protective clothing should be either disposed properly or deposited for laundering by the institution. Laboratory protective clothing should not be taken home. c. Eye and face protection (goggles, mask, face-shield or other splatter guards) should be used for anticipated splashes or sprays of infectious or other hazardous materials when the microorganisms must be manipulated outside the BSC or containment device. d. Gloves must be worn to protect hands from exposure to hazardous material. (1) Gloves must not be worn outside the laboratory. (2) Gloves selection should be based on an appropriate risk assessment. (3) Alternates to latex gloves should be available. (4) Gloves should be changed when contaminated, integrity has been compromised, or when otherwise necessary. (5) Wearing two pairs of gloves may be appropriate. (6) Gloves should be removed and hands should be washed when work with hazardous materials has been completed and before leaving the laboratory. (7) Disposable gloves should not be washed or reused. (8) Gloves should be disposed with other contaminated laboratory waste. e. Hand washing protocols must be rigorously followed. f.

Eye, face, and respiratory protection should be used in rooms containing infected animals as determined by the risk assessment.

4. Laboratory Facilities (Secondary Barriers) (BSL-2) a. Laboratory doors should be self-closing and have locks in accordance with the institutional policies.

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b. 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. c. The laboratory should be designed so that it can be easily cleaned and decontaminated. Carpets and rugs in laboratories are not permitted. d. Laboratory furniture must be capable of supporting anticipated loading and uses. Spaces between benches, cabinets, and equipment are accessible for cleaning. (1) Bench tops must be impervious to water and resistant to moderate heat, organic solvents, acids, alkalis, and other chemicals (such as those used to decontaminate the work surfaces and equipment). (2) Chairs and other furniture used in laboratory work should be covered with a nonporous material that can be easily cleaned and decontaminated with appropriate disinfectant. e. 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. f.

BSCs must be installed in such a manner that fluctuations of the room supply and exhaust air do not interfere with proper operations. BSCs should be located away from doors, windows that can be opened, heavily traveled laboratory areas, and other potentially disruptive equipment.

g. Vacuum lines should be protected with High Efficiency Particulate Air (HEPA) filters, or their equivalent. Filters must be replaced as needed. Liquid disinfectant traps may be required. h. An eyewash station must be readily available. i.

There are no specific requirements on ventilation systems.

j.

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.

k. 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 a direct (hard) connection. Provisions to assure proper safety cabinet performance and air system operation must be verified. l.

A method for decontaminating all laboratory wastes should be available in the facility (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method). End section for BSL-2

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E. Biological Safety Level 3 (BSL-3) Biosafety Level 3 (BSL-3) is applicable to clinical, diagnostic, teaching, research, or production facilities in which work is performed with indigenous or exotic agents which may cause serious or potentially lethal disease through inhalation route exposure. Laboratory personnel must receive specific training in handling pathogenic and potentially lethal agents, and must be supervised by scientists competent in handling infectious agents and associated procedures. All procedures involving the manipulation of infectious materials are conducted within BSCs or other physical containment devices, or by personnel wearing appropriate personal protective equipment. The laboratory has special engineering and design features. UCO does not have a facility that will meet the BSL-3 criteria at this time. The following standard and special safety practices, equipment and facilities apply to agents assigned to BSL-3.

1. Standard Microbiological Practices (BSL-3) a. Access to the laboratory is limited or restricted when experiments are in progress. b. Persons must wash their hands after working with potentially hazardous materials, after removing gloves, and before leaving the laboratory. c. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption is not permitted in the laboratory. Food must be stored outside the work area in cabinets or refrigerators designated and used for this purpose only. d. Mouth pipetting is prohibited; mechanical pipetting devices must be used. e. Policies for the safe handling of sharps such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. Whenever practical, PIs 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: (1) Hypodermic needles and syringes should only be used for parenteral injection and aspiration of fluids from laboratory animals or diaphragm bottles. Only needle-locking syringes or disposable syringe-needle unites (i.,e., needle is integral to the syringe) may be used for the injection or aspiration of fluids containing microorganisms or viruses. (2) Safe needle devices should be used whenever such a device is available for the task requiring the use of a needle/syringe. (3) 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 (4) Used disposable needles and syringes must be carefully placed in conveniently located puncture-resistant containers used for sharps disposal.

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(5) Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. (6) Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. Plasticware should be substituted for glassware whenever possible. f.

All procedures should be performed carefully to minimize the creation of splashes and/or aerosols.

g. Work surfaces should be decontaminated after completion of work and after any spill or splash of potentially infectious material with an appropriate disinfectant. h. All cultures, stocks, and other potentially infectious materials should be decontaminated before disposal using an effective method. A method for decontaminating all laboratory wastes should be available in the facility, preferably within the laboratory (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method). (1) Materials to be decontaminated outside of the immediate laboratory must be placed in a durable, leak-proof container and secured for transport. (2) Materials to be removed from the facility for decontamination must be packaged in accordance with applicable local, state, and federal regulations. i.

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.

j.

An effective integrated pest management program is required.

k. The PI must ensure that laboratory personnel receive appropriate training regarding their duties, the necessary precautions to prevent exposures, and exposure evaluation procedures. l.

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 child-bearing age should be provided with information regarding immune competence and conditions that may predispose them to infection.

m. Individuals having these conditions should be encouraged to self-identify to Employee Health for appropriate counseling and guidance.

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2. Special Practices (BSL-3) a. Laboratory doors are kept closed when experiments are in progress. b. The laboratory director controls access to the laboratory and restricts access to persons whose presence is required for program or support purposes. The laboratory director establishes policies and procedures whereby only persons who have been advised of the potential biohazard, who meet any specific entry requirements (e.g., immunization), and who comply with all entry and exit procedures, enter the laboratory or animal rooms. c. Laboratory personnel are provided medical surveillance and offered appropriate immunizations or tests for the agents handled or potentially present in the laboratory, and any periodic testing as recommended for the agent being handled. d. Baseline serum samples are collected and stored for all laboratory and other at-risk personnel when such procedures are recommended by the PI and/or the Institutional Biosafety Committee. Additional serum specimens may be periodically collected, depending on the agents handled or the function of the laboratory. e. A laboratory-specific biosafety manual must be prepared and adopted as policy. The biosafety manual must be available and accessible. Personnel are required to read and to follow these practices and procedures. f.

Laboratory and support personnel receive appropriate training on the potential hazards associated with the work involved, the necessary precautions to prevent exposures, and the exposure evaluation procedures. Personnel receive annual updates, or additional training as necessary for procedural changes.

g. The PI must ensure that laboratory personnel demonstrate proficiency in standard and special microbiological practices and techniques and in the practices and operations specific to the laboratory facility, before working with BSL-3 agents. h. Potentially infectious materials must be placed in a durable, leak proof container during collection, handling, processing, storage, or transport within the facility. i. Laboratory equipment should be routinely decontaminated and decontaminated after spills, splashes, or other potential contamination. j.

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

k. Equipment must be decontaminated before repair, maintenance, or removal from the laboratory. l.

Incidents that may result in exposure to infectious materials must be immediately evaluated and treated according to procedures described in the laboratory biosafety safety manual. All such incidents must be reported to the PI and to the IBC. Medical evaluation, surveillance, and treatment should be provided through Employee Health or the nearest emergency room and appropriate records maintained.

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m. Animals and plants not associated with the work being performed must not be permitted in the laboratory. n. All procedures involving the manipulation of infectious materials must be conducted within a BSC, or other physical containment devices. No work with open vessels is conducted on the bench. When a procedure cannot be performed within a BSC, a combination of personal protective equipment and other containment devices, such as a centrifuge safety cup or sealed rotor, must be used.

3. Safety Equipment (Primary Barriers) (BSL-3) a. All procedures involving the manipulation of infectious materials must be conducted within a BSC (preferably Class II or Class III), or other physical containment devices. b. Protective laboratory clothing with a solid-front such as a tie-back or wraparound gowns, scrub suits, or coveralls are worn by workers when in the laboratory. Protective clothing is not worn outside the laboratory. Reusable clothing is decontaminated before being laundered. Clothing is changed when contaminated. c. 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. Eye and face protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse. Persons who wear contact lenses in laboratories must also wear eye protection. d. Gloves must be worn to protect hands from exposure to hazardous material. (1) Gloves must not be worn outside the laboratory. (2) Gloves selection should be based on an appropriate risk assessment. (3) Alternates to latex gloves should be available. (4) Gloves should be changed when contaminated, integrity has been compromised, or when otherwise necessary. (5) Wearing two pairs of gloves may be appropriate. (6) Gloves should be removed and hands should be washed when work with hazardous materials has been completed and before leaving the laboratory. (7) Disposable gloves should not be washed or reused. (8) Gloves should be disposed with other contaminated laboratory waste. e. Hand washing protocols must be rigorously followed. f.

Eye, face, and respiratory protection should be used in rooms containing infected animals.

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4. Laboratory Facilities (Secondary Barriers) (BSL-3) a. Laboratory doors must be self-closing and have locks in accordance with the institutional policies. The laboratory must be separated from areas that are open to unrestricted traffic flow within the building. Access to the laboratory is restricted to entry by a series of two self-closing doors. A clothing change room (anteroom) may be included in the passageway between the two self-closing doors. b. Laboratories must contain a sink for hand washing. The sink must be hands-free or automatically operated. It should be located near the exit door. If the laboratory is segregated into different laboratories, a sink must also be available for hand washing in each zone. Additional sinks may be required as determined by the risk assessment. c. The laboratory must be designed so that it can be easily cleaned and decontaminated. Carpets and rugs are not permitted. Seams, floors, walls, and ceiling surfaces should be sealed. Spaces around doors and ventilation openings should be capable of being sealed to facilitate space decontamination. (1) Floors must be slip-resistant, impermeable to liquids, and resistant to chemicals. Consideration should be given to the installation of seamless, sealed, resilient or poured floors, with integral cove bases. (2) Walls should be constructed to produce a sealed smooth finish that can be easily cleaned and decontaminated. (3) Ceilings should be constructed, sealed, and finished in the same general manner as walls. d. Decontamination of the entire laboratory should be considered when there has been gross contamination of the space, significant changes in laboratory usage, for major renovations, or maintenance shut downs. Selection of the appropriate materials and methods used to decontaminate the laboratory must be based on the risk assessment of the biological agents in use. e. Laboratory furniture must be capable of supporting anticipated loads and uses. Spaces between benches, cabinets, and equipment must be accessible for cleaning. (1) Bench tops must be impervious to water and resistant to heat, organic solvents, acids, alkalis, and other chemicals. (2) Chairs used in laboratory work must be covered with a non-porous material that can be easily cleaned and decontaminated with appropriate disinfectant. f.

All windows in the laboratory must be sealed.

g. 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, heavily traveled laboratory areas, and other possible airflow disruptions.

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h. Vacuum lines must be protected with HEPA filters, or their equivalent. Filters must be replaced as needed. Liquid disinfectant traps may be required. i.

An eyewash station must be readily available in the laboratory.

j.

A ducted air ventilation system is required. This system must provide sustained directional airflow by drawing air into the laboratory from “clean” areas toward “conditions the airflow will not be reversed. (1) Laboratory personnel must be able to verify directional air flow. A visual monitoring device which confirms directional air flow must be provided at the laboratory entry. Audible alarms should be considered to notify personnel of air flow disruption. (2) The laboratory exhaust air must not re-circulate to any other area of the building. (3) The laboratory building exhaust air should be dispersed away from occupied areas and from building air intake locations or the exhaust air must be HEPA filtered.

k. HEPA filtered exhaust air from a Class II BSC can be safely re-circulated 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 a direct (hard) connection. Provisions to assure proper safety cabinet performance and air system operation must be verified. BSCs should be certified at least annually to assure correct performance. Class III BSCs must be directly (hard) connected up through the second exhaust HEPA filter of the cabinet. Supply air must be provided in such a manner that prevents positive pressurization of the cabinet. l.

A method for decontaminating all laboratory wastes should be available in the facility, preferably within the laboratory (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method).

m. Equipment that may produce infectious aerosols must be contained in devices that exhaust air through HEPA filtration or other equivalent technology before being discharged into the laboratory. These HEPA filters should be tested and/or replaced at least annually. n. Facility design consideration should be given to means of decontaminating large pieces of equipment before removal from the laboratory. o. Enhanced environmental and personal protection may be required by the agent summary statement, risk assessment, or applicable local, state, or federal regulations. These laboratory enhancements may include, for example, one or more of the following; an anteroom for clean storage of equipment and potentially contaminated” areas. The laboratory shall be designed such that under failure supplies with dress-in, shower-out capabilities; gas tight dampers to facilitate laboratory isolation; final HEPA filtration of the laboratory exhaust air; laboratory effluent decontamination; and advanced access control devices such as biometrics. HEPA filter housings should have gas-tight isolation dampers; decontamination ports; and/or bag-in/bag-out (with appropriate decontamination procedures) capability. The HEPA filter housing should allow for leak

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testing of each filter and assembly. The filters and the housing should be certified at least annually. p. The BSL-3 facility design, operational parameters, and procedures must be verified and documented prior to operation. Facilities must be re-verified and documented at least annually. End BSL-3 section

F. Biological Safety Level 4 Biosafety Level 4 (BSL-4) is required for work with dangerous and exotic agents which pose a high individual risk of aerosol-transmitted laboratory infections and life-threatening disease. At this time, the University of Central Oklahoma does not have a facility designed for work with organisms at BSL-4.

G. Animal Biosafety Level 1 (ABSL-1) Animal Biosafety Level 1 (ABSL-1) is suitable for work involving well characterized agents that are not known to cause disease in immune-competent adult humans, and present minimal potential hazard to personnel and the environment.

1. Standard Practices (ABSL-1) a. The animal facility director establishes policies, procedures, and protocols for emergency situations. b. Prior to any project’s initiation, each protocol must be submitted for review and approval by the Institutional Animal Care and Use Committee (IACUC) and the Institutional Biosafety Committee (IBC). Any special practices are approved at this time. c. A safety manual specific to the animal facility should be prepared and be available and accessible. Personnel are advised of potential hazards, and are required to read and follow instructions on practices and procedures. d. Supervisors must ensure that animal care, laboratory and support personnel receive appropriate training regarding their duties, animal husbandry procedures, 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. e. All employees working with animals must participate in the medical surveillance program through Employee Health, and must complete and submit the Animal Handler’s Questionnaire to Employee Health. 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 child-bearing age should be provided information regarding immune competence and conditions that may predispose them to

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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. f.

A sign incorporating safety information must be posted at the entrance to the areas where infectious materials and/or animals are housed or are manipulated. The sign must include the animal biosafety level, general occupational health requirements, personal protective equipment requirements, the PI name (or other responsible personnel), telephone number, and required procedures for entering and exiting the animal areas. Identification of specific infectious agents is recommended when more than one agent is being used within an animal room.

g. Access to the animal room is limited. Only those persons required for program or support purposes are authorized to enter the facility. All persons, including facility personnel, service workers, and visitors are advised of the potential hazards (nature of research pathogens, allergens, etc.) and are instructed on the appropriate safeguards. h. Protective laboratory coats, gowns, or uniforms are required 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 minimizes 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 and face and respiratory protection should be used in rooms containing infected animals, as dictated by the risk assessment. i.

Eating, drinking, smoking, handling contact lenses, applying designated areas and are not permitted in animal or procedure rooms.

j.

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

k. Mouth pipetting is prohibited. Mechanical pipetting devices must be used. l.

Policies for the safe handling of sharps such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. (1) When applicable, PIs should adopt improved engineering and work practice controls that reduce the risk of sharps injuries, including the use of safe needle devices wherever possible. (2) Needles and syringes or other sharp instruments should be limited to use in the animal facility when there is no alternative for such procedures as parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles.

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BIOLOGICAL SAFETY MANUAL i. Disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal. ii. 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. iii. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving.

(3) Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. (4) Plasticware should be substituted for glassware whenever possible. (5) Equipment containing sharp edges and corners should be avoided. m. Equipment and work surfaces should be routinely decontaminated with an appropriate disinfectant after work with an infectious agent and after spills, splashes, or overt contamination. n. 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 are manipulated. o. An effective integrated pest management program is required. p. All wastes from the animal room (including animal tissues, carcasses, and contaminated bedding) are transported from the animal room in leak-proof, covered containers for appropriate disposal. Incineration is recommended.

2. Special Practices: None required. (ABSL-1) 3. Safety Equipment (Primary Barriers and Personal Protective Equipment) (ABSL-1) a. A risk assessment should be performed by the PI to determine the appropriate type of personal protective equipment to be utilized. b. The wearing of laboratory coats, gowns, or uniforms are recommended to prevent contamination of personal clothing. c. Protective outer clothing such as laboratory coats should not be worn outside areas where infectious materials and/or animal are housed or manipulated. Gowns and uniforms are not worn outside the facility. d. Protective eyewear should be worn when conducting procedures that have the potential to create splashes of microorganisms or other hazardous materials. Persons who wear

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contact lenses should also wear eye protection when entering areas with potentially high concentrations or airborne particulates. e. Persons having contact with non-human primates should assess their risk of mucous membrane exposure and wear appropriate eye and face protection. f.

Gloves should be 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. (1) Gloves should be changed when contaminated, the integrity has been compromised, or when otherwise necessary. (2) Gloves must not be worn outside the animal rooms. (3) Gloves and personal protective equipment should be removed in a manner that prohibits transfer of infectious materials. (4) Disposable gloves should not be washed or reused. Dispose of used gloves with other contaminated waste.

g. 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.

4. Animal Facilities (Secondary Barriers) (ABSL-1) a. The animal facility is separated from areas that are open to unrestricted personnel traffic within the building. b. External facility doors are self-closing and self-locking. Access to the animal facility is restricted. 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. c. The animal facility must have a sink for hand washing. Sink traps are filled with water, and/or appropriate liquid to prevent the migration of vermin and gases. d. The animal facility is designed, constructed, and maintained to facilitate cleaning and housekeeping. The interior surfaces (walls, floors, and ceilings) are water resistant. e. The animal facility is designed, constructed, and maintained to facilitate cleaning and housekeeping. The interior surfaces (walls, floors, and ceilings) are water resistant. It is recommended that penetrations in floors, walls and ceiling surfaces are sealed, to include openings around ducts, doors and door frames, to facilitate pest control and proper cleaning. Floors must be slip resistant, impervious to liquids, and resistant to chemicals.

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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. 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.

g. External windows are not recommended. Any windows must be resistant to breakage. Where possible, windows should be sealed. If the animal facility has windows that open, they should be fitted with fly screens. h. Ventilation should be provided in accordance with the Guide for Care and Use of Laboratory Animals, latest edition. No recirculation of exhaust air should occur. It is recommended that animal rooms have inward directional airflow. Ventilation system design should consider the heat and high moisture load produced during the cleaning of animal rooms and the cage wash process. i.

Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, should be arranged to minimize horizontal surface areas to facilitate cleaning and minimize the accumulation of debris or fomites.

j.

If floor drains are provided, the traps must always be filled with water and/or an appropriate disinfectant to prevent the migration of vermin and gases.

k. Cages should be washed manually, or preferably, in a mechanical cage washer. The mechanical cage washer should have a final rinse temperature of at least 180 º F. l. Illumination should be adequate for all activities, avoiding reflections and glare that could impede vision. m. Emergency eyewash and shower should be readily available; location is determined by the risk assessment.

H. Animal Biological Safety Level 2 (ABSL-2) Animal Biosafety Level 2 involves practices for work with those agents associated with human disease. It addresses hazards from ingestion as well as from percutaneous and mucous membrane exposure. ABSL-2 builds upon the practices, procedures, containment equipment, and facility requirements of ABSL-1.

1. Standard Practices (ABSL-2) a. Standard policies, procedures, and protocols for emergency situations should be established by the facility director. Appropriate special standard operating procedures must be developed by the PI and reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) and the Institutional Biosafety Committee (IBC). The form is located at (http://www.uco .pdf) will need a site for forms and policies may be used to facilitate this process.

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(1) All personnel working in the animal room must be advised of special hazards, and are required to read and follow these SOPs. Personnel must be supervised by individuals with adequate knowledge of potential hazards, microbiological agents, animal manipulations, and husbandry procedures. (2) Animal care, laboratory and support personnel receive appropriate training regarding their duties, animal husbandry procedures, potential hazards associated with the work involved, manipulations of infectious agents, the necessary precautions to prevent exposures, and the hazard/exposure evaluation procedures (physical hazards, splashes, aerosolization, etc.). (3) Personnel should receive annual updates, or additional training as necessary for procedural or policy changes. (4) Records of all training provided should be maintained. b. All employees working with animals must participate in the medical surveillance program through Employee Health, and must complete and submit the Animal Handler’s Questionnaire to Employee Health. 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 child-bearing 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. c. A sign incorporating the universal biohazard symbol must be posted at the entrance to the 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 requirements, the PI name (or other responsible personnel), telephone number, and required procedures for entering and exiting the animal areas. Identification of specific infectious agents is recommended when more than one agent is being used within an animal room. d. Access to the animal room should be limited. Only those persons required for program or support purposes should be 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 hazard (natural or research pathogens, allergens, etc.) and are instructed on the appropriate safeguards. e. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use should only be done in designated areas and is not permitted in animal or procedure rooms. f.

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

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g. Mouth pipetting is prohibited. Mechanical pipetting devices must be used. h. Policies for the safe handling of sharps such as needles, scalpels, pipettes, and broken glassware must be developed and implemented. (1) When applicable, PIs should adopt improved engineering and work practice controls that reduce the risk of sharps injuries, including the use of safe needle devices wherever possible. (2) Needles and syringes or other sharp instruments are limited to use in the animal facility when there is no alternative for such procedures as parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. i. Disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal. ii. 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. iii. Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. (3) Broken glassware must not be handled directly. Instead, it must be removed using a brush and dustpan, tongs, or forceps. (4) Plasticwareshould be substituted for glassware whenever possible. (5) Equipment containing sharp edges and corners should be avoided. i.

Equipment and work surfaces in the room should be routinely decontaminated with an effective disinfectant after work with the infectious agent, and after spills, splashes, or other overt contamination.

j.

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 are manipulated.

k. An effective integrated pest management program is required. l.

All wastes from the animal room (including animal tissues, carcasses, and contaminated bedding) should be transported from the animal room in leak-proof, covered containers for appropriate disposal. Incineration is recommended.

2. Special Practices (ABSL-2) a. Procedures involving a high potential for generating aerosols (e.g., necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, or intranasal inoculation of animals) should be conducted within a biosafety cabinet (BSC) or other physical containment device. When a procedure cannot be performed within a BSC, a

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combination of personal protective equipment and other containment devices must be used (e.g., respirators, face shields, glove boxes). Consideration should be given to the use of restraint devices and practices that reduce the risk of exposure during animal manipulations (e.g., physical restraint devices, chemical restraint medications, etc). b. All infectious samples should be collected, labeled, transported, and processed in a manner that contains and prevents transmission of the agent(s). All potentially infectious materials and wastes from the animal room (including animal tissues, carcasses, contaminated bedding, unused feed, sharps, and other refuse) should be transported from the animal room in leak-proof, covered containers for appropriate disposal (autoclaving or incineration). (1) A means for decontaminating routine husbandry equipment, sensitive electronic and medical equipment should be developed and used. (2) 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. (2) The outer surface of the container is disinfected prior to moving materials. (3) The transport container must contain a universal biohazard label. (4) The outer surface of the containers must be disinfected prior to moving the material. (5) Equipment, cages, and racks should be handled in 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. c. Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material. d. Incidents that may result in exposure to infectious materials must be immediately evaluated and treated through Employee Health (or the Emergency Room, after hours). All such incidents must be reported to the PI and the IBC. Medical evaluation, surveillance, and treatment should be provided through Employee Health or the nearest emergency room and records maintained.

3. Safety Equipment (Primary Barriers and Personal Protective Equipment) a. Properly maintained BSCs, personal protective equipment (e.g., gloves, lab coats, face shields, respirators, etc.) and/or other physical containment devices or equipment, should be used whenever conducting procedures with a potential for creating aerosols or splashes. These include necropsy of infected animals, harvesting of tissues or fluids from infected animals or eggs, and intranasal inoculation of animals. b. 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 larger cages placed in inward

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flow ventilated enclosures or other equivalent primary containment systems for larger animal cages. c. A risk assessment should determine the appropriate type of personal protective equipment to be utilized. d. Protective laboratory coats, gowns, scrub suites, or uniforms should be worn while in the areas where infectious materials and/or animals are housed or manipulated to prevent contamination of personal clothing. This protective clothing should be removed prior to leaving the animal facility. (1) Reusable clothing should be appropriately contained and decontaminated before being laundered. (2) Laboratory and protective clothing should never be taken home. (3) Disposable personal protective equipment and other contaminated waste are appropriately contained and decontaminated prior to disposal. e. Gloves should be worn to prevent skin contact with contaminated, infectious and hazardous materials and when handling animals. A risk assessment should be performed to identify the appropriate glove for the task, and alternatives to latex gloves should be available. (1) Gloves should be changed when contaminated, the integrity has been compromised, or when otherwise necessary. (2) Gloves must not be worn outside the animal rooms. (3) Gloves and personal protective equipment should be removed in a manner that prohibits transfer of infectious materials. (4) Disposable gloves should not be washed or reused. (5) Dispose of used gloves with other contaminated waste. f.

Eye and face protection (mask, goggles, face shield or other splatter guard) are used for anticipated splashes/ sprays from infectious or other hazardous materials and when the animal or microorganisms must be handled outside the BSC or containment device as described in the SOPs. (1) Protective eyewear should be worn when conducting procedures that have the potential to create splashes of microorganisms or other hazardous materials. Persons who wear contact lenses should also wear eye protection when entering areas with potentially high concentrations or airborne particulates. (2) Persons having contact with non-human primates should assess their risk of mucous membrane exposure and wear appropriate eye and face protection.

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g. Respiratory protection should be worn when dictated by the risk assessment and identified in the SOPs. h. Gloves and personal protective equipment should be removed in a manner that minimizes transfer of infectious materials outside of the areas where infectious materials and/or animals are housed or are manipulated. i.

Eye, face and respiratory protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse.

j.

Persons must wash their hands after removing gloves, and before leaving the areas where infectious materials and/or animals are housed or are manipulated.

4. Animal Facilities (Secondary Barriers) (ABSL-2) a. The animal facility is separated from areas that are open to unrestricted personnel traffic within the building. External doors are self-closing and self-locking. b. Access to the facility is limited by secure locked doors. Doors, to areas where infectious materials and/or animal 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. c. A hand washing sink is located at the exit of the areas where infectious materials and/or animals are housed or are manipulated. If the animal facility has segregated areas where infectious materials and/or animals are housed or manipulated, a sink must be available for hand washing at the exit from each segregated area. d. Sink traps are filled with water and/or appropriate liquid to prevent the migration of vermin and gasses. e. 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, to include openings around ducts, doors and door frames, to facilitate pest control and proper cleaning. Floors must be slip resistant, impervious to liquids, and resistant to chemicals. f.

Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, are arranged to minimize horizontal surface areas.

g. 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. h. External windows are not recommended. Any windows must be sealed and resistant to breakage.

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i.

Ventilation should be provided in accordance with criteria from Guide for Care and Use of Laboratory Animals, latest edition. The direction of airflow in the animal facility is inward; animal rooms should maintain inward directional airflow compared to adjoining hallways. A ducted exhaust air ventilation system is required, and exhaust air is discharged to the outside without being re-circulated 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.

j.

Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, should be arranged to minimize horizontal surface areas, to facilitate cleaning and minimize the accumulation of debris or fomites.

k. If floor drains are provided, the traps should always be filled with water or an appropriate disinfectant to prevent the migration of vermin and gasses. l.

Cages should be autoclaved or otherwise decontaminated prior to washing. The mechanical cage washer should have a final rinse temperature of at least 180 ºF. The cage wash area should be designed to accommodate the use of high pressure spray systems; humidity, strong chemical disinfectants and 180 ºF water temperatures, during the cage/equipment cleaning process.

m. Illumination should be adequate for all activities, avoiding reflections and glare that could impede vision. n.

If BSCs are present, they must be installed so that fluctuations of the room air supply and exhaust do not interfere with proper operations. (1) 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. (2) BSCs can also be connected to the laboratory exhaust system by either a thimble (canopy) connection or a direct (hard) connection. (3) Provisions to assure proper safety cabinet performance and air system operation must be verified. Correct performance of the BSCs should be recertified at least once a year. All BSCs should be used according to manufacturer’s recommendation, to protect the worker and avoid creating a hazardous environment from volatile chemical and gases.

o. If vacuum service (i.e., central or local) is provided, each service connection should be fitted with liquid disinfectant traps and an inline HEPA filter, placed as near as practicable to each use point or service cock. Filters should be installed to permit inplace decontamination and replacement. p. Availability of an autoclave in the animal facility should be considered to facilitate decontamination of infectious materials and waste.

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q. Emergency eyewash and shower should be readily available; with the location determined by the risk assessment.

I. Animal Biological Safety Level 3 (ABSL-3) Animal Biosafety Level 3 involves practices suitable for work with animals infected with indigenous or exotic agents that present the potential of aerosol transmission and of causing serious or potentially lethal disease. ABSL-3 builds upon the standard practices, procedures, containment equipment, and facility requirements of ABSL-2.

1. Standard Practices (ABSL-3) a. Policies, procedures, and protocols for routine work and emergency situations must be established by the facility director and the PI, including worker safety and health concerns, which are reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) and the Institutional Biosafety Committee (IBC) prior to beginning a study. b. A safety manual specific to the animal facility must be prepared in consultation with the animal facility director and the Environmental Health and Safety Office. The safety manual must be available and accessible. Personnel must be advised on the potential and special hazards, and are required to read and follow instructions on practices and procedures. c. The PI 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.). (1) Personnel must receive annual updates or additional training when procedures or policies change. (2) Records must be maintained for all hazard evaluations, employee training sessions and staff attendance. d. All employees working with animals must participate in the medical surveillance program through Employee Health, and must complete and submit the Animal Handler’s Questionnaire to Employee Health. All personnel should be offered appropriate immunizations or tests for the agents handled or potentially present. 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 child-bearing 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.

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e. Personnel using respirators must be enrolled in the UCO respiratory protection program. Safety Operations Manager will handle all fit tests and training for employees. f.

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. The sign must include the animal biosafety level, general occupational health requirements, personal protective equipment requirements, the PI’s name (or other responsible personnel), telephone number, and required procedures for entering and exiting the animal areas. Identification of specific infectious agents is recommended when more than one agent is being used within an animal room. Security-sensitive agent information and occupational health requirements should be reviewed by the UCO IBC prior to posting.

g. Advance consideration should be given to emergency and disaster recovery plans, as a contingency for man-made or natural disasters. h. Access to the animal room must be limited to the fewest number of individuals possible. 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 are manipulated. All persons including facility personnel, service workers, and visitors must be advised of the potential hazards (natural or research pathogens, allergens, etc) and must be instructed on the appropriate safeguards. i.

Gloves and personal protective equipment should be worn in the animal area and removed in a manner that minimizes transfer of infectious materials outside of the areas where infectious materials and/or animals are housed or are manipulated. (1) Protective laboratory coats, gowns, or uniforms are required to prevent contamination of personal clothing. (2) Gloves should be worn to prevent skin contact with contaminated, infectious and/or hazardous materials and when handling animals. Double-glove practices should be used when dictated by risk assessment. (3) Eye and face and respiratory protection should be used in rooms containing infected animals, as dictated by the risk assessment.

j.

Persons must wash their hands after removing gloves, and before leaving the areas where infectious materials and/or animals are housed or are manipulated.

k. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human use should only be done in designated areas and are not permitted in animal or procedure rooms. l.

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

m. Mouth pipetting is prohibited. Mechanical pipetting devices must be used. m. Policies for the safe handling of sharps, such as needles, scalpels, pipettes, and broken glassware must be developed and implemented, including the following.

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(1) When applicable, PIs should adopt improved engineering and work practice controls that reduce the risk of sharps injuries. (2) Needles and syringes or other sharp instruments are limited to use in the animal facility when there is no alternative for such procedures as parenteral injection, blood collection, or aspiration of fluids from laboratory animals and diaphragm bottles. (3) 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. (4) Non-disposable sharps must be placed in a hard-walled container for transport to a processing area for decontamination, preferably by autoclaving. (5) Broken glassware must not be handled directly; it should be removed using a brush and dustpan, tongs, or forceps. Plasticware should be substituted for glassware whenever possible. (6) Equipment containing sharp edges and corners should be avoided. n. 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. o. 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 are manipulated. p. An effective integrated pest management program is required. q. All wastes from the animal room (including animal tissues, carcasses, and bedding) must be autoclaved prior to removal from the animal facility and transported from the animal room in leak proof containers for disposal by incineration through the biomedical waste disposal company on contract with the University.

2. Special Practices (ABSL-3) a. 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 baseline serum sample should be stored. b. All procedures involving the manipulation of infectious materials, handling infected animals or the generations of aerosols must be conducted within BSCs or other physical containment devices when practical. When a procedure cannot be performed within a biosafety cabinet, a combination of personal protective equipment and other containment devices must be used.

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c. Consideration should be given to the use of restraint devices and practices that reduce the risk of exposure during animal manipulations (e.g., physical restraint devices, chemical restraint medications, etc). d. The risk of infectious aerosols from infected animals or their bedding also can be reduced if animals are housed in containment caging systems (such as solid wall and bottom cages covered with filter bonnets, open cages placed in inward flow ventilated enclosures, HEPA-filter isolators and caging systems, or other equivalent primary containment systems). e. Actively ventilated caging systems must be designed to prevent the escape of microorganisms from the cage. Exhaust plenums for these systems should be sealed to prevent escape of microorganisms if the ventilation system becomes static, and the exhaust must be HEPA filtered. Safety mechanisms should be in place that prevent the cages and exhaust plenums from becoming positive to the surrounding area should the exhaust fan fail. The system should also be alarmed to indicate when operational malfunctions occur. f.

A method for decontaminating all infectious materials must be available within the facility, preferably within the areas where infectious materials and/or animals are housed or are manipulated (e.g. autoclave, chemical disinfection, or other approved decontamination methods). Consideration should be given to means for decontaminating routine husbandry equipment, sensitive electronic and medical equipment.

g. All potential infectious materials (including animal tissues, carcasses, contaminated bedding, unused feed, sharps, and other refuse) must be autoclaved before removal from the areas where infectious materials and/or animals are housed or are manipulated by an appropriate method. It is recommended that animal bedding and waste be decontaminated prior to manipulation and before removal from the areas where infectious materials and/or animals are housed or are manipulated, preferably within the caging system. h. Equipment, cages, and racks should be handled in 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. i.

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

j.

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 director, the IACUC and the IBC.

k. Medical evaluation, surveillance, and treatment should be provided through Employee Health or the nearest emergency room, and records maintained.

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3. Safety Equipment (Primary Barriers and P.P.E.) (ABSL-3) a. Properly maintained BSCs, and other physical containment devices or equipment, should be used for all manipulations for infectious materials and when possible, animals. These manipulations include necropsy, harvesting of tissues or fluids from infected animals or eggs, and intranasal inoculation of animals. b. The risk of infectious aerosols from infected animals or bedding can be reduced through the use of primary barrier systems. c. These systems may include solid wall and bottom cages covered with filter bonnets; ventilated cage rack systems; or for larger cages placed in inward flow ventilated enclosures or other equivalent systems or devices. d. A risk assessment should determine the appropriate type of protective clothing and personal protective equipment to be utilized. (1) Protective clothing such as uniforms or scrub suits should be worn by personnel within the animal facility. i. Reusable clothing should be appropriately contained and decontaminated before being laundered. Laboratory and protective clothing should never be taken home. ii. Disposable personal protective equipment such as non-woven olefin cover-all suits, wrap-around or solid-front gowns should be worn over this clothing, before entering the areas where infectious materials and/or animals are housed or manipulated. Front-button laboratory coats are unsuitable. (2) Appropriate eye, face and respiratory protection should be worn by all personnel entering areas where infectious materials and/or animals are housed or are manipulated. i. Persons who wear contact lenses should also wear eye protection when entering areas with potentially high concentrations or airborne particulates. ii. Eye and face protection must be disposed of with other contaminated laboratory waste or decontaminated before reuse. (3) To prevent cross contamination boots, shoe covers, or other protective footwear, should be used where indicated. (4) Gloves should be 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. i. Procedures may require the use of wearing two pairs of gloves (doubleglove). ii. Gloves must be changed when contaminated, integrity has been compromised, or when otherwise necessary. iii. Gloves must not be worn outside the animal rooms.

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BIOLOGICAL SAFETY MANUAL iv. Gloves and personal protective equipment should be removed in a manner that prohibits transfer of infectious materials. Do not wash or reuse disposable gloves. Dispose of used gloves with other contaminated waste.

(5) Disposable personal protective equipment must be removed when leaving the areas where infectious materials and/or animals are housed or are manipulated. i. Scrub suits and uniforms should be removed before leaving the animal facility. ii. Disposable personal protective equipment and other contaminated waste are appropriately contained and decontaminated prior to disposal. e. 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.

4. Laboratory Facilities (Secondary Barriers) (ABSL-3) a. The animal facility must be separated from areas that are open to unrestricted personnel traffic within the building. External facility doors are self-closing and self-locking. b. Access to the animal facility is restricted. Doors to areas where infectious materials and/or animals are housed or 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. c. Entry into the containment area is via a double-door entry which constitutes an anteroom/airlock and a change room. (1) Showers may be considered based on the risk assessment. (2) An additional double-door access anteroom or double-door autoclave may be provided for movement of supplies and wastes into and out of the facility. d. A hand washing sink is located at the exit of the areas where infectious materials and/or animals are housed or are manipulated. The sink should be hands-free or automatically operated. (1) Additional sinks for hand washing should be located in other appropriate locations within the facility. (2) If the animal facility has multiple segregated areas where infectious materials and/or animals are housed or are manipulated, a sink must also be available for hand washing at the exit from each segregated area. (3) Sink traps are filled with water, and/or appropriate liquid to prevent the migration of vermin and gases.

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e. The animal facility should be designed, constructed, and maintained to facilitate cleaning, decontamination and housekeeping. The interior surfaces (walls, floors and ceilings) should be water resistant. f.

Penetrations in floors, walls and ceiling surfaces are sealed, to include openings around ducts, doors and door frames, to facilitate pest control, proper cleaning and decontamination. Walls, floors and ceilings should form a sealed and surfaces should be easily sanitized.

g. Floors must be slip resistant, impervious to liquids, and resistant to chemicals. Flooring should be seamless, sealed resilient or poured floors, with integral cove bases. h. Decontamination of an entire animal room should be considered when there has been gross contamination of the space, significant changes in usage, for major renovations, or maintenance shut downs. Selection of the appropriate materials and methods used to decontaminate the animal room must be based on the risk assessment. i.

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.

j.

Furniture should be minimized. Chairs used in animal area must be covered with a nonporous 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.

k. External windows are not recommended; if present, all windows must be sealed and must be resistant to breakage. The presence of windows may impact facility security and therefore should be assessed by security personnel. l.

Ventilation to the facility should be provided in accordance with the Guide for Care and Use of Laboratory Animals. The direction of airflow into the animal facility should be inward; animal rooms should maintain inward directional airflow compared to adjoining hallways. A ducted exhaust air ventilation system should be provided. Exhaust air should be discharged to the outside without being recirculated to other rooms. This system creates directional airflow which draws air into the animal room from "clean" areas and toward "contaminated" areas.

m. Ventilation system design should consider the heat and high moisture load produced during the cleaning of animal rooms and the cage wash process. Filtration and other treatments of the exhaust air may not be required, but should be considered based on site requirements, specific agent manipulations and use conditions. The exhaust must be dispersed away from occupied areas and air intakes, or the exhaust must be HEPAfiltered. n. Personnel must verify that the direction of the airflow (into the animal areas) is proper. It is recommended that a visual monitoring device that indicates and confirms directional inward airflow be provided at the animal room entry.

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o. Consideration should be given to installing an HVAC control system to prevent sustained positive pressurization of the animal spaces. Audible alarms should be considered to notify personnel of ventilation and HVAC system failure. p. Internal facility appurtenances, such as light fixtures, air ducts, and utility pipes, should be arranged to minimize horizontal surface areas, to facilitate cleaning and minimize the accumulation of debris or fomites. q. Floor drains must be maintained and filled with water, and/or appropriate disinfectant to prevent the migration of vermin and gases. r.

Cages must be washed in a mechanical cage washer. The mechanical cage washer has a final rinse temperature of at least 180oF. Cages should be autoclaved or otherwise decontaminated prior to removal from ABSL-3 space. The cage wash facility should be designed and constructed to accommodate high pressure spray systems, humidity, strong chemical disinfectants and 180oF water temperatures, during the cage cleaning process.

s. Illumination should be adequate for all activities, avoiding reflections and glare that could impede vision. (1) BSCs (Class II, Class III) must be installed so that fluctuations of the room air supply and exhaust do not interfere with its proper operations. Class II BSCs should be located away from doors, heavily traveled laboratory areas, and other possible airflow disruptions. (2) 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. (3) BSCs can also be connected to the laboratory exhaust system by either a thimble (canopy) connection or a direct (hard) connection. Provisions to assure proper safety cabinet performance and air system operation must be verified. (4) Class III BSCs must supply air in such a manner that prevents positive pressurization of the cabinet or the laboratory room. t.

BSCs should be certified at least annually to assure correct performance.

u. BSCs should be used according to manufacturers’ recommendations. v. When applicable, equipment that may produce infectious aerosols must be contained in devices that exhaust air through HEPA filtration or other equivalent technology before being discharged into the animal facility. These HEPA filters should be tested and/or replaced at least annually. w. An autoclave must be available which is convenient to the animal rooms where the biohazard is contained. The autoclave is utilized to decontaminate infectious materials and waste before moving it to the other areas of the facility. If not convenient to areas where infectious materials and/or animals are housed or are manipulated, special

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practices should be developed for transport of infectious materials designated alternate location/s within the facility. x. An emergency eyewash and shower must be readily available. y. Additional environmental protection (e.g., personnel showers, HEPA filtration of exhaust air, containment of other piped services, and the provision or effluent decontamination) should be considered if recommended by the agent summary statement, as determined by risk assessment of the site conditions, or other applicable federal, state or local regulations. z. The ABSL-3 facility design and operational procedures must be documented. aa. (1) The facility must be tested to verify that the design and operational parameters have been met prior to use. (2) Facilities should be re-verified at least annually against these procedures as modified by operational experience.

J. Animal Biological Safety Level 4 (ABSL-4) Animal Biosafety Level 4 (ABSL-4) involves practices suitable for addressing dangerous or exotic agents that pose high risk of life threatening disease, aerosol transmission, or related agents with unknown risk of transmission. At this time, the University of Central Oklahoma does not have a facility designed for ABSL-4 work.

K. Guidelines for Work with Toxins of Biological Origin 1. Each laboratory worker must be trained in the theory and practice of the toxins to be used, with special emphasis on the nature of the practical hazards associated with laboratory operations. This includes how to handle transfers of liquids containing toxin, where to place waste solutions and contaminated materials or equipment, and how to decontaminate work areas after routine operations, as well as after accidental spills. The worker must be reliable and sufficiently adept at all required manipulations before being provided with toxin. 2. A risk assessment should be conducted to develop safe operating procedures before undertaking laboratory operations with toxins. If toxins and infectious agents are used together, then both must be considered when containment equipment is selected and safety procedures are developed. Likewise, animal safety practices must be considered for toxin work involving animals. 3. Each laboratory that uses toxins should develop standard operating procedures specific for the toxin(s) to be used. 4. An inventory control system should be in place to account for toxin use and disposition. If toxins are stored in the laboratory, containers should be sealed, labeled, and secured

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to ensure restricted access; refrigerators and other storage containers should be clearly labeled and provide contact information for trained, responsible laboratory staff. 5. Laboratory work with toxins should be done only in designated rooms with controlled access and at pre-determined bench areas. When toxins are in use, the room should be clearly posted: “Toxins in Use-Authorized Personnel Only.” Unrelated and nonessential work should be restricted from areas where stock solutions of toxin or organisms producing toxin are used. Visitors or other untrained personnel granted laboratory access must be monitored and protected from inadvertently handling laboratory equipment used to manipulate the toxin or organism. 6. Routine operations with dilute toxin solutions should be conducted under BSL-2 conditions with the aid of personal protective equipment and a well maintained BSC or comparable engineering controls. Engineering controls should be selected according to the risk assessment for each specific toxin operation. A certified BSC or chemical fume hood will suffice for routine operations with most protein toxins. Low molecular weight toxin solutions, or work involving volatile chemicals or radio-nucleotides combined with toxin solutions, may require the use of a charcoal-based hood filter in addition to HEPA filtration. All work with toxins should be conducted within the operationally effective zone of the hood or BSC, and each user should verify the inward airflow before initiating work. 7. When using an open-fronted fume hood or BSC, workers should wear suitable laboratory PPE to protect the hands and arms, such as laboratory coats, smocks, or coveralls and disposable gloves. When working with toxins that pose direct percutaneous hazards, special care must be taken to select gloves that are impervious to the toxin and the diluents or solvents employed. 8. When conducting liquid transfers and other operations that pose a potential splash or droplet hazard in an open-fronted hood or BSC, safety glasses and disposable N-95 respirator and/or a face shield, should be worn. 9. Toxin should be removed from the hood or BSC only after the exterior of the closed primary container has been decontaminated and placed in a clean secondary container. Toxin solutions, especially concentrated stock solutions, should be transported in leak/spill-proof secondary containers. 10. The interior of the hood or BSC should be decontaminated periodically, for example, at the end of a series of related experiments. Until thoroughly decontaminated, the hood or BSC should be posted to indicate that toxins remain in use, and access should remain restricted. 11. Selected operations with toxins may require modified BSL-3 practices and procedures. The determination to use BSL-3 should be made in consultation with the Institutional Biosafety Committee and is based upon a risk assessment that considers the variables of each specific laboratory operation, especially the toxin under study, the physical state of the toxin (solution or dry form), the total amount of toxin used relative to the estimated human lethal dose, the volume of the material manipulated, the methodology, and any human or equipment performance limitations. 12. Emphasis must be placed on evaluating and modifying experimental procedures to eliminate the possibility of inadvertent generation of toxin aerosols.

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a. Pressurized tubes or other containers holding toxins should be opened in a BSC, chemical fume hood, or other ventilated enclosure. b. Operations that expose toxin solutions to vacuum or pressure, for example sterilization of toxin solutions by membrane filtration, should always be handled in this manner, and the operator should also use appropriate respiratory protection. c. If vacuum lines are used with toxin, they should be protected with a HEPA filter to prevent entry of toxins into the line. d. Centrifugation of cultures or materials potentially containing toxins should only be performed using sealed, thick-walled tubes in safety centrifuge cups or sealed rotors. The outside surfaces of containers and rotors should be routinely cleaned before each use to prevent contamination that may generate an aerosol. After centrifugation, the entire rotor assembly is taken from the centrifuge to a BSC to open it and remove its tubes. 13. Accidental needle-sticks or mechanical injury from “sharps” such as glass or metal implements pose a well-known risk to laboratory workers, and the consequences may be catastrophic for operations involving toxins in amounts that exceed a human lethal dose. a. Only workers trained and experienced in handling animals should be permitted to conduct operations involving injection of toxin solutions using hollow-bore needles. b. Discarded needles/syringes and other sharps should be placed directly into properly labeled, puncture-resistant sharps containers, and decontaminated as soon as is practical. c. Glassware should be replaced with plastic for handling toxin solutions wherever practical to minimize the risk of cuts or abrasions from contaminated surfaces. d. Thin-walled glass equipment should be completely avoided. Glass Pasteur pipettes are particularly dangerous for transferring toxin solutions and should be replaced with disposable plastic pipettes. Glass chromatography columns under pressure must be enclosed within a plastic water jacket or other secondary container. e. Experiments should be planned to eliminate or minimize work with dry toxin (e.g. freeze-dried preparations). Unavoidable operations with dry toxin should only be undertaken with appropriate respiratory protection and engineering controls. Dry toxin can be manipulated using a Class III BSC, or with the use of secondary containment such as a disposable glove bag or glove box within a hood or Class II BSC. “Static-free” disposable gloves should be worn when working with dry forms of toxins that are subject to spread by electrostatic dispersal. (1) In specialized laboratories, the intentional, controlled generation of aerosols from toxin solutions may be undertaken to test antidotes or vaccines in experimental animals. These are extremely hazardous operations that should

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BIOLOGICAL SAFETY MANUAL only be conducted after extensive validation of equipment and personnel, using non-toxic simulants. (2) Aerosol exposure of animals should be done in a certified Class III BSC or hood line. While removing exposed animals from the hood line, and for required animal handling during the first 24 hours after exposure, workers should take additional precautions, including wearing protective clothing (e.g., disposable Tyvek suit) and appropriate respiratory protection. To minimize the risk of dry toxin generating a secondary aerosol, areas of animal skin or fur exposed to aerosols should be gently wiped with a damp cloth containing water or buffered cleaning solution before the animals are returned to holding areas. (3) For high-risk operations involving dry forms of toxins, intentional aerosol formation, or the use of hollow-bore needles in conjunction with amounts of toxin estimated to be lethal for humans, consideration should be given to requiring the presence of at least two knowledgeable individuals at all times in the laboratory. f.

Procedures for toxin inactivation should be established and followed. (1) Depending upon the toxin, contaminated materials and toxin waste solutions can be inactivated by incineration or extensive autoclaving, or by soaking in suitable decontamination solutions. (2) All disposable material used for toxin work should be placed in secondary containers, autoclaved and disposed of as toxic waste. (3) Contaminated or potentially contaminated protective clothing and equipment should be decontaminated using suitable chemical methods or autoclaving before removal from the laboratory for disposal, cleaning or repair. If decontamination is impracticable, materials should be disposed of as hazardous or bio-hazardous waste, in consultation with the Environmental Health and Safety Office.

g. In the event of a spill, avoid splashes or generating aerosols during cleanup by covering the spill with paper towels or other disposable, absorbent material. Apply an appropriate decontamination solution to the spill, beginning at the perimeter and working towards the center, and allow sufficient contact time to completely inactivate the toxin. Sources: CDC/NIH Biosafety in Microbiological and Biomedical Laboratories, U.S. Department of Health and Human Services Public Health Services, Centers for Disease Control and Prevention and National Institutes of Health, HHS Publication No. (CDC) 93-8395, 4th. Edition, May, 1999, and 5th. Edition, Feb., 2007. NIH Guidelines for Research Involving Recombinant DNA Molecules, April, 2002.

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L. Procedures for Working with Human Blood or Other Potentially Infectious Material 1. Departments with employees who have reasonably anticipated eye, skin, mucous membrane or parenteral contact with human blood or other potentially infectious materials must follow the UCO Exposure Control Plan in compliance with the OSHA Bloodborne Pathogen Standard. 2.

Other potentially infectious material means the following: a. Human Body Fluids: (1) Semen, (2) Vaginal secretions, (3) Pericardial fluid, (4) Cerebrospinal fluid, (5) Synovial fluid, (6) Pleural fluid, (7) Pericardial fluid, (8) Peritoneal fluid, (9) Amniotic fluid, (10) Saliva in dental procedures, (11) any body fluid that is visibly contaminated with blood, (12) all body fluids in situations where it is difficult or impossible to differentiate between body fluids, b. Other: (1) any unfixed tissue or organ (other than intact skin) from a human, living or dead, (2) human immunodeficiency virus (HIV)-containing cell or tissue cultures, (3) human organ cultures, (4) HIV or hepatitis B virus (HBV) containing culture medium or other solutions, and (5) Blood, organs, or other tissues from experimental animals infected with HIV, HBV or other bloodborne pathogens infectious to man.

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c. Special Procedures for Human Tissue/Cells/Cell Lines: (1) Only established human cell lines and human cell strains which are characterized (tested by antigenic screening for viral or agent markers, co-cultivation with indicator cells allowing contaminants to grow, or molecular technology such as polymerase chain reaction or nucleic acid hybridization) to be free of bloodborne pathogens (including HIV, HBV, Epstein-Barr virus, Herpes virus and papilloma members of the Papovavirus group, etc.) and documented as such may be excluded from the requirements of the OSHA Bloodborne Pathogen Standard. (2) Cell lines/strains that are procured from commercial vendors or other sources with documented testing to be free of human bloodborne pathogens and which have been protected from contamination may be excluded from the requirements of the OSHA Bloodborne Pathogen Standard. (3) Use of human tissue/cells/cell lines in the laboratory setting requires the approval of the UCO Institutional Review Board. 3. Universal precautions should be observed which dictates that all human blood and other potentially infectious materials should be treated as infectious for HBV, HIV, and other bloodborne pathogens. 4. Engineering and work practice controls should be utilized first to minimize employee exposure. Where occupational exposure remains after the institution of engineering controls, PPE should also be used as follows: a. Gloves should be worn when it can be reasonably anticipated that the employee may have hand contact with blood or other potentially infectious materials, mucous membranes, and non-intact skin such as during phlebotomies and when handling or touching contaminated items. b. Masks in combination with eye protection devices such as goggles or face shields should be worn whenever splashes, spray, spatter, or droplets of blood or other potentially infectious material may be generated and eye, nose, or mouth contamination can be reasonably anticipated. c. Gowns, aprons, lab coats, surgical caps or hoods, and/or shoe covers should be worn when gross contamination can be reasonably anticipated. The type and characteristics of this protective clothing will depend upon the task and degree of exposure anticipated. 5. Hand washing facilities should be readily accessible to employees. Personnel in work areas that do not have hand washing facilities readily accessible should be provided with an appropriate hand cleanser in conjunction with clean cloth or paper towels or antiseptic towelettes. Employees should wash their hands with soap and running water as soon as feasible after using antiseptic hand cleansers or towelettes. 6. All garments should be removed as soon as possible if penetrated by blood or other potentially infectious material.

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7. Removed PPE should be placed in a designated area or container for storage, washing, decontamination, or disposal. Contaminated PPE should be placed in a designated container labeled with the biohazard symbol. a. PPE should be cleaned, laundered and/or disposed in a proper manner. (1) Contaminated disposable PPE should be placed in a biohazard bag until it can be sterilized / autoclaved. (2) Contaminated PPE, that can be laundered, should be placed in a container labeled with the biohazard symbol until sent to an appropriate laundry in accordance with the UCO’s Exposure Control Plan. 8. Employees should wash their hands immediately or as soon as feasible after removal of gloves or other PPE. 9. Employees should wash their hands or other skin with soap and water or flush mucous membranes with water immediately or as soon as feasible following contact of such body areas with blood or other potentially infectious materials. 10. Safe needle devices should be used where possible or appropriate. 11. Contaminated needles or other contaminated sharps should not be bent, recapped, or removed. If needles must be recapped, a mechanical means or a one-handed technique should be used. 12. Immediately or as soon as possible after use, contaminated sharps should be placed in appropriate containers, even if the sharps are reusable and will be reprocessed. a. These containers should be: (1) puncture resistant, (2) labeled with the biohazard symbol or color-coded, (3) leak-proof on the sides and bottom, and (4) not be allowed to overfill (a good guideline is to dispose when approximately twothirds full). b. Other guidelines for selection of sharps containers should consider issues such as lids that lock tight for safe disposal, a container that is specifically constructed for the method of sterilization that will be used (if sharps containers are not specifically constructed to be autoclaved, the resulting mass of melted plastic is extremely hazardous due to the needles that often protrude), and a clear top that would allow inspection. 13. Eating, drinking, smoking, applying cosmetics or lip balm and handling contact lenses is prohibited in areas where there is a reasonable likelihood of occupational exposure. 14. Food or drink should not be kept in areas where blood or other potentially infectious materials are present or stored.

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15. Procedures which minimize spraying, splashing, spattering, and generation of droplets of infectious material shall be used whenever possible. 16. No mouth pipetting should occur. 17. Biohazard labels should be affixed to all containers of regulated waste, refrigerators, freezers and other containers that hold or are contaminated with blood or other potentially infectious material. Red bags or containers may be substituted for labels. 18. Specimens of blood or other potentially infectious materials should be placed in a container which prevents leakage during collection, storage, transport, or shipping. This container should be red or labeled with the biohazard symbol and closed prior to being stored, transported, or shipped. If contamination outside this primary container occurs or is likely to occur, it should be placed in a second red or similarly labeled container which prevents leakage during handling processing, storage, transport, or shipping. 19. Equipment which has been in contact with blood or other potentially infected material should be examined prior to servicing or shipping and should be decontaminated as necessary. a. Where complete decontamination cannot occur prior to servicing, a readily observable biohazard label shall be attached to the equipment stating which portions of the equipment remains contaminated, and b. the employee requesting the service or repair is responsible for ensuring that information is conveyed to all affected employees, service representatives such as the UCO Biomed Vendor and/or the manufacturer prior to handling, servicing, or shipping so that appropriate precautions can be taken. 20. Contaminated work surfaces should be decontaminated after completion of procedures, immediately or as soon as feasible after any spill of blood or other potentially infectious material and at the end of the work shift if the surface has become contaminated since the last cleaning. 21. Broken glassware which may be contaminated should not be picked up directly with the hands but by mechanical means such as a brush and dustpan, tongs or forceps. 22. All employees with occupational exposure should receive bloodborne pathogen training at the time of assignment to tasks where occupational exposure may take place, when changes affect employees' occupational exposure and at least annually thereafter. 23. The hepatitis B vaccine should be made available to all employees who have occupational exposure to blood or other potentially infectious materials. 24. If an employee sustains an exposure incident (such as a stick with a contaminated needle/scalpel/dental wire or a splash of potentially infectious material in the eye, mouth, mucous membrane, or non-intact skin), the exposed person should immediately: a. clean the wound with soap; flush mucous membranes with water or normal saline solution;

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b. notify his/her supervisor, designated coordinator, or other designated individual; c. proceed for treatment within 1-2 hours of the exposure (see the UCO Infectious Diseases Policy for current recommended treatment locations); and d. if possible, for laboratory exposures, bring a sample of the source material to the treatment facility for testing. Sources: OSHA Bloodborne Pathogens Standard (29 CFR 1910.1030

M. Recombinant DNA The National Institutes of Health (NIH) publishes Guidelines for Research Involving Recombinant DNA Molecules in the Federal Register which should be followed when constructing and handling recombinant DNA molecules or organisms and viruses containing recombinant DNA molecules. The most current version of this document is available upon request from the EHS Office. UNDER THESE GUIDELINES, THE PRINCIPAL INVESTIGATOR MUST: 1. Make an initial determination of the required levels of physical and biological containment in accordance with the NIH Guidelines; 2. Select appropriate microbiological practices and laboratory techniques to be used for the research; 3. Submit the initial research protocol and any subsequent changes (e.g., changes in the source of DNA or host-vector system) to the Institutional Biosafety Committee (IBC) for review and approval or disapproval; 4. Remain in communication with the IBC throughout the conduct of the project; 5. Make available to all laboratory staff the protocols that describe the potential biohazards and the precautions to be taken; 6. Instruct and train laboratory staff in: a. the practices and techniques required to ensure safety, and b. the procedures for dealing with accidents; 7. Inform the laboratory staff of the reasons and provisions for any precautionary medical practices advised or requested (e.g., vaccinations or serum collection); 8. Supervise the safety performance of the laboratory staff to ensure that the required safety practices and techniques are employed;

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9. Investigate and report any significant problems pertaining to the operation and implementation of containment practices and procedures in writing to the Environmental Health and Safety Office (EHSO), Animal Facility Director (where applicable), IBC, NIH/OBA, and other appropriate authorities (if applicable) (reports to NIH/OBA shall be sent to the Office of 10. Biotechnology Activities, National Institutes of Health, 6705 Rockledge Drive, Suite 750, MSC 7985, Bethesda, MD 20892-7985 (20817 for non- USPS mail), 301-496-9838, 301496-9839 (fax); 11. Correct work errors and conditions that may result in the release of recombinant DNA materials; 12. Ensure the integrity of the physical containment (e.g., BSCs) and the biological containment (e.g., purity and genotypic and phenotypic characteristics); and 13. Comply with reporting requirements for human gene transfer experiments conducted in compliance with the NIH Guidelines. Source: NIH Guidelines for Research Involving Recombinant DNA Molecules, April, 2002.

N. Institutional Biological Safety Committee The UCO Institutional Biosafety Committee (IBC) has the charge of reviewing and approving all research and clinical research activities involving recombinant DNA, gene transfer, microorganisms and biological toxins. PRIOR TO THE COMMENCEMENT OF ANY PROJECT INVOLVING THE USE OF ANY OF THESE MATERIALS, THE PRINCIPLE INVESTIGATOR MUST PERFORM THE FOLLOWING STEPS. 1. Review the applicable guidelines and regulations and become familiar with the safety procedures and requirements, including: a. The National Institutes of Health (NIH) Guidelines for Research Involving Recombinant DNA Molecules; b. The Center for Disease Control and Prevention (CDC) and NIH publication Biosafety in Microbiological and Biomedical Labs; c. 32 CFR Part 626 Biological Defense Safety Program and 32 CFR Part 627 Biological Defense Safety Program, Technical Safety Requirements (DA Pamphlet 385-69); d. The Occupational Safety and Health Administration (OSHA) Bloodborne Pathogens Standard; and e. The OSHA Occupational Exposure to Hazardous Chemicals in Laboratories Standard. 2. Perform a risk assessment of the agents and procedures to determine potential safety and environmental hazards.

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3. Develop laboratory specific standard operating procedures (SOPs) to address such issues as: a. the hazards; b. the risk classification of the agent; c. immunizations or other medical considerations required; d. employee training on the specific hazards and procedures to be performed in the lab; e. access control if applicable; f.

personal protective equipment (type of glove, lab coats, goggles, etc.) and safety equipment (BSC, chemical fume hood, etc.) requirements for defined tasks to be performed;

g. decontamination/cleaning requirements (frequency, disinfection agent, etc.) of all potentially contaminated surfaces; h. storage requirements (location, container, access, whether prior approval is required from the Principle Investigator to obtain, etc.); i.

disposal/inactivation procedures for any and all contaminated items such as the organism/agent, gloves, pipettes, etc.;

j.

spill procedures (specific, including who to notify, where to go for medical attention and forms to complete afterward);

k. employee exposure and other emergency procedures (what to do first, who to call, where to go); and l.

posting of emergency contact numbers outside of the laboratory.

4. Complete and sign the appropriate form(s): a. IBC Protocol Review/Approval Form - for grants/contracts involving the use of recombinant DNA, any microorganism or virus, and any biological toxin, including those considered by the CDC to be a “select agent” requiring registration with the CDC. For microorganism use requiring BSL-2 procedures or higher, the SOPs that have been developed must be attached. b. IBC Human Protocol Approval Form: Gene Transfer, Microorganism/Virus/Toxin Administration - for grants/contracts/ clinical trials involving the use of human gene transfer (the transfer of recombinant DNA molecules into one or more human research participants) or microorganism, virus or toxin administration to humans. c. Form for Resubmission of an IBC Approved Project or its Submission to a Different Funding Source - if the grant/contract is a resubmission to the same or an alternate

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funding source and the previously approved protocol remains unchanged, this shortened form should be submitted. If changes in any other item(s) occurs, such as organisms used, biosafety level or NIH classification, a completed IBC Protocol Review/Approval Form or IBC Human Protocol Approval Form: Gene Transfer, Microorganism/ Virus/Toxin Administration must be submitted. d. Transgenic/Knockout Vertebrate Animal Submission Form – this form is for grants/contracts involving the generation of animals in which the animal's genome will be altered by stable introduction of recombinant DNA, or DNA derived there from, into the germ-line or modified to eliminate specific gene(s) (transgenic/knockout animals) where such animals are created in an UCO facility. This form is also required for the mating of two transgenic lines to create a double transgenic. This form is not required for the purchase and/or use of transgenic vertebrate animals when these animals are obtained from a source outside UCO. 5. Submit the signed form(s) to the Institutional Biosafety Committee Office. Investigators may be contacted by the IBC for additional information or clarification regarding the research project. Once approved, the Principle Investigator will receive a letter from the IBC indicating approval and an IBC approval number.

O. Animal Safety 1. Zoonotic Diseases A zoonotic disease is defined as one that is communicable from animals to humans. Some species posing risks (in decreasing order of hazard) are provided in Table VI-1. Strategies for staying healthy and reducing the risk of exposure are as follows. a. Wash your hands frequently - the most common method of contracting a zoonotic infection is by placing infectious material in your mouth, nose, or eyes. b. Wear personal protective clothing and equipment (see Table VI-1) and do not take unlaundered protective clothing home. c. Follow recommended safe work practices (see Tables VI-2 and VI- 3). d. Notify your supervisor immediately if an exposure incident occurs, even if it seems minor, and then seek medical attention. First-aid kits are readily available in all buildings housing animals, then report to a medical facility such as McBride Clinic. e. Tell your personal physician that you work with animals. Many zoonotic diseases have flu-like symptoms: your physician needs this information for making an accurate diagnosis.

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2. Injury Environmental factors, as well as factors intrinsic to the animal, can lead to greater risk for injury. Animals respond to sounds and smells, sometimes undetectable to humans, which can frighten the animal. Animals may have a flight zone or a particular sign of distress that animal handlers should be aware of to reduce risk. Inappropriate handling can induce discomfort, pain and distress, provoking an animal to inflict injury on its handler. Animals, especially nonhuman primates, may grab or get caught in loose clothing, long hair, etc., or may spit or throw feces. Guidelines for preventing injury include: a. Know the animal’s flight zone and signs of distress. b. Use proper handling techniques. c. Minimize the use of sharps and glass and ensure proper disposal or same. d. Determine the potential risk and wear appropriate protective equipment for the hazard, which may include leather gloves, latex/nitrile gloves, face shield, etc. e. If you must lift heavy objects, contact the EHSO for safe lifting procedures and training. f.

Minimize the amount of time a floor is allowed to remain wet, and use slip-resistant footwear, mats and signage whenever wet floors cannot be avoided.

g. If a bite or a scratch is sustained, it is important that medical care be sought, even if the injury seems trivial, due to the potential for disease transmission, which can include Cercopithecine herpes virus 1 (B-virus), Rabies, hantavirus infection, cat-scratch\ fever, tularemia, rat-bite fever, and Staphylococcus infection. Procedures to follow in the event of a bite or scratch are: (1) While wearing gloves, carefully express the wound and apply gentle pressure around the wound to encourage bleeding. (2) Rinse the wound under warm running water for 15 minutes and continue massaging the site. (3) If the injury involves a macaque or any animal known to be infected with a zoonotic disease, you may wash the wound and surrounding area with povidone-iodine solution for 5 minutes. Care should be taken when using iodine, as prolonged skin exposure to iodine can increase wound healing time and can cause tissue damage. (4) For all other wounds, or if you are concerned about using iodine, wash the wound and surrounding area with soap and water for 5 minutes. (5) Continue to rinse periodically. If normal saline is available, rinse with normal saline. (6) Pat the injury dry using sterile gauze pads. (7) Cover the wound with a pad and secure it with gauze and tape.

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(8) Seek medical attention.

3. Animal Allergies/Asthma Animals and animal products such as dander, hair, scales, fur, saliva, and body wastes contain powerful allergens that can cause both respiratory and skin disorders. Sources of exposure to animal allergens vary with animal species. For example, allergens have been found in the urine of rats; the urine, saliva, and pelts of guinea pigs; rabbit pelts, cat saliva and dander; dog dander; and horse serum and dander. Inhalation is one way for animal allergens to enter the body. After a period of time (often several months, but occasionally many years), one may inhale sufficient quantities of allergens to become sensitized - that is, develop symptoms when exposed again, even to tiny amounts of the allergen. Other routes of exposures may come from animal bites or scratches. In sensitized persons, reactions often occur soon after exposure to the animal or animal product, but they may be delayed for 2 to 8 hours or more. Symptoms vary from mild reactions such as sneezing and runny nose to more serious reactions such as cough, chest tightness, wheezing, or shortness of breath. The National Institute for Occupational Safety and Health (NIOSH) recommends several measures to reduce exposures to animal allergens in the workplace and prevent animal-induced asthma and allergies, including the following: a. Provide training to educate workers about animal allergies and steps for risk reduction. b. Perform animal manipulations within ventilated hoods or safety cabinets when possible. c. Avoid wearing street clothes while working with animals. Leave work clothes at the workplace to avoid potential exposure problems for family members. d. Keep cages and animal areas clean. Take particular care to control exposures during cleaning (minimize dust and aerosols, etc.). e. Reduce skin contact with animal products such as dander, serum, and urine by using gloves, lab coats, and approved particulate respirators with faceshields where appropriate. f.

Provide health monitoring and appropriate counseling and medical follow-up for workers who have become sensitized or have developed allergy symptoms.

4. Institutional Animal Care and Use Committee (IACUC) Requirements The UCO IACUC is responsible for the overview of animal research at UCO, and has determined that annual training in both animal safety and animal handling procedures is mandatory for all UCO personnel working with animals. Anyone who has a concern about their care, tending, and/or use of research animals should report the matter directly to either the Animal Resources PI, the IACUC chair, or any IACUC

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committee member. These concerns, and any action taken as a results, will be presented to the IACUC chair and then to the entire IACUC committee. Each facility or program that uses animals will be inspected by the IACUC at least once every six months. Sources: Biosafety in the Laboratory: Prudent Practices for the Handling and Disposal of Infectious Materials, National Research Council, National Academy Press, 1989. Biosafety in Microbiological and Biomedical Laboratories, fourth edition, Centers for Disease Control and Prevention/National Institutes of Health, U. S. Government Printing Office, 1999. NIOSH ALERT: Preventing Asthma in Animal Handlers, DHHS (NIOSH) Publication No. 97-116, January 1998. Occupational Health and Safety in the Care and Use of Research Animals, National Research Council, National Academy Press, 1997. OUHSC Biosafety Manual

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TABLE VI-1 DISEASES, PROTECTIVE EQUIPMENT, AND MEDICAL MONITORING/VACCINATIONS FOR CERTAIN ANIMAL SPECIES Species

Potential Zoonotic Disease

Non-human Primates

Circopithecine herpes virus 1: CHV 1 (Herpes virus simiae) Hepatitis A Shigellosis (Shigella spp.) Campylobacteriosis (Campylobacter spp.)

Dogs

Campylobacteriosis Leptospirosis (Leptospira interrogans) Salmonellosis (Salmonella spp.) Rabies

Cats

Toxoplasmosis (Toxoplasma gondii) Cat-scratch fever (Bartonella henselae) Salmonellosis Rabies Microsporosis/Ringworm (Epidermophyton floccosum, Microsporum spp., Trichophyton spp.)

Sheep

Q fever (Coxiella bunettii)

Minimum Personal Protective Equipment Disposable surgical face mask Disposable gloves Disposable shoe covers Disposable hair bonnet Disposable gown Face shield** Gloves appropriate for the hazard (leather to protect against bites, latex/nitrile to protect against biological material) Laboratory coat Face protection (mask, goggles, face shield) when potential for splash of hazardous material exists Gloves appropriate for the hazard (leather to protect against bites, latex/nitrile to protect against biological material) Laboratory coat Face protection (mask, goggles, face shield) when potential for splash of hazardous material exists Gloves appropriate for the hazard (leather to protect against bites, latex/nitrile to protect against biological

* Medical Monitoring/Vaccinations Baseline and annual physical Serum banking (recommended) Tetanus immunization TB test (every 6 months) Hepatitis B immunization (recommended for those working with apes) Baseline and annual physical Serum banking (recommended) Tetanus immunization Rabies vaccination

Baseline and annual physical Serum banking (recommended) Tetanus immunization Rabies vaccination Toxoplasmosis antibodies titer for premenopausal personnel

Baseline and annual physical Serum banking (recommended) Tetanus immunization Q-Fever assessment

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material) Laboratory coat Face protection (mask, goggles, face shield) when potential for splash of hazardous material exists Rabbits Leptospirosis Gloves appropriate for the Baseline and annual physical Microsporosis/Ringworm hazard (leather to protect Serum banking (recommended) against bites, latex/nitrile to Tetanus immunization protect against biological material) Laboratory coat Face protection (mask, goggles, face shield) when potential for splash of hazardous material exists Laboratory Leptospirosis Gloves appropriate for the Baseline and annual physical raised Salmonellosis hazard (leather to protect Serum banking (recommended) rodents Microsporosis against bites, latex/nitrile to Tetanus immunization Ringworm protect against biological Allergies material) Laboratory coat Face protection (mask, goggles, face shield) when potential for splash of hazardous material exists *For routine work where the animal is not known to be infected with an organism for research purposes. Where infection is known, the appropriate Animal Biosafety Level precautions shall be used (see Table VI-3). **Persons performing cage cleaning, moving, or handling of a conscious animal must wear additional protective equipment.

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TABLE VI-2 RECOMMENDED SAFE WORK PRACTICES Practices to Reduce the Number of Employees at Risk of Exposure

• •

• • Practices to Reduce Exposures by Direct and Indirect Contact

Practices to Reduce Percutaneous Exposures

Practices to Reduce Exposure by Ingestion

• • • • • • • • • • • • • •

Practices to Reduce Exposure by Inhalation

• •

Restrict access to the work area. Provide warnings of hazards (such as biohazards or chemical hazards) and advice about special requirements (such as personal protective equipment or immunization requirements). Keep hands away from mouth, nose, eyes, and skin. Wash hands when contaminated and when work activity is completed; especially after handling animals and before leaving the work area. Decontaminate work surfaces before and after work and after spills of a hazardous agent. Use appropriate methods to decontaminate equipment, surfaces, and wastes. Substitute less-hazardous materials for hazardous materials whenever possible. Wear personal protective equipment while performing work and remove it before leaving the work area. Eliminate the use of sharp objects whenever possible. Use needles with self-storing sheaths or those designed to protect the user. Select products with puncture-resistant features whenever possible. Use puncture-resistant sharps containers for disposal of sharps. Handle animals with care and proper restraint to prevent scratches and bites. No mouth pipetting allowed. Do not smoke, eat, drink, or apply cosmetics in areas used for the care and use of research animals. Keep hands and contaminated items away from mouth. Protect mouth from splash and splatter hazards. Use chemical fume hoods, BSCs, and other containment equipment to control inhalation hazards. Handle fluids carefully to avoid spills and splashes and the generation of aerosols. Use in-line HEPA filters to protect the vacuum system.

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TABLE VI-2 RECOMMENDED SAFE WORK PRACTICES Practices to Reduce the Number of Employees at Risk of Exposure

• •

• • Practices to Reduce Exposures by Direct and Indirect Contact

Practices to Reduce Percutaneous Exposures

Practices to Reduce Exposure by Ingestion

• • • • • • • • • • • • • •

Practices to Reduce Exposure by Inhalation

• •

Restrict access to the work area. Provide warnings of hazards (such as biohazards or chemical hazards) and advice about special requirements (such as personal protective equipment or immunization requirements). Keep hands away from mouth, nose, eyes, and skin. Wash hands when contaminated and when work activity is completed; especially after handling animals and before leaving the work area. Decontaminate work surfaces before and after work and after spills of a hazardous agent. Use appropriate methods to decontaminate equipment, surfaces, and wastes. Substitute less-hazardous materials for hazardous materials whenever possible. Wear personal protective equipment while performing work and remove it before leaving the work area. Eliminate the use of sharp objects whenever possible. Use needles with self-storing sheaths or those designed to protect the user. Select products with puncture-resistant features whenever possible. Use puncture-resistant sharps containers for disposal of sharps. Handle animals with care and proper restraint to prevent scratches and bites. No mouth pipetting allowed. Do not smoke, eat, drink, or apply cosmetics in areas used for the care and use of research animals. Keep hands and contaminated items away from mouth. Protect mouth from splash and splatter hazards. Use chemical fume hoods, BSCs, and other containment equipment to control inhalation hazards. Handle fluids carefully to avoid spills and splashes and the generation of aerosols. Use in-line HEPA filters to protect the vacuum system.

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TABLE VI-3 ADDITIONAL PRECAUTIONS FOR ANIMAL BIOSAFETY LEVELS 2 AND 3 • • Biosafety Level 2

• • • • • • • •

Biosafety Level 3

• • • • •

Written standard operating procedures must be developed. Specific training must be provided to laboratory and animal handlers regarding the infectious agent and special procedures to be used. This training must be documented. Only animals used for the experiment should be allowed in the room. Personal protective equipment may need to be disposable as biomedical waste or require decontamination before reuse. Floor drain traps should be filled with an appropriate disinfectant Waste should be evaluated to determine whether there is a need to autoclave prior to disposal. Transport of animals should only occur in a manner that does not cause transmission of infectious materials. Additional immunization or medical monitoring may be warranted depending on the agent. All Biosafety Level 2 procedures. Personal protective equipment must include double gloves, eye and face protection, wrap-around or solid-front gowns (front-button laboratory coats are unsuitable) and respiratory protection, and must be disposed as biomedical waste or decontaminated before removal from the room. All wastes must be decontaminated/autoclaved prior to disposal Cages are autoclaved or thoroughly decontaminated before being cleaned or washed. A spill procedure is developed an posted. Materials related to the experiment are not permitted in the animal room.

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