EH&S: Biosafety Manual - Principles of Biosafety

Each laboratory conducting BSL 1 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory.

BSL 1 work, by definition, involves agents of no known hazard or of low potential hazard to personnel and the environment. BSL 1 practices, safety equipment and facilities are appropriate for most undergraduate training and teaching laboratories, and for other facilities in which work is done with defined and characterized strains of viable microorganisms not known to cause disease in healthy adult humans.

BSL 1 represents a basic level of containment that relies on standard microbiological practices with no special primary or secondary barriers recommended, other than a sink for handwashing. The laboratory is not separated from the general traffic patterns in the building. Work is generally conducted on open bench tops. Special containment devices or equipment such as a biological safety cabinet are generally not required for manipulations of agents assigned to BSL 1. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or related science.

a. Persons wash their hands after they handle viable materials, after removing gloves and before leaving the laboratory.

b. Eating, drinking, smoking, handling contact lenses, applying cosmetics and storing food for human use are not permitted in the work areas. Persons who wear contact lenses in laboratories should also wear goggles or a face shield. Food is stored outside the work area in cabinets or refrigerators designated and used for this purpose only.

c. Mechanical pipetting devices are used; pipetting by mouth is strictly prohibited.

d. Policies for the safe handling of sharps are instituted.

e. A biohazard sign can be posted at the entrance to the laboratory whenever infectious agents are present. The sign may include the name of the agent(s) in use and the name and telephone number of the investigator.

f. An insect and rodent control program is in effect. (Example: If the laboratory has windows that open, they are fitted with fly screens).

a. The laboratory is designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate.

b. Bench tops are impervious to water and resistant to acids, alkalis, organic solvents, moderate heat and chemicals used to decontaminate the work surface and equipment.

c. Laboratory furniture is sturdy. Spaces between benches, cabinets and equipment are accessible for cleaning.

The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 1 agents and those who work in or enter a BSL 1 facility. The practices and procedures set forth here are intended to augment the BSL 1 and ABSL 1 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment.

a. The PI must ensure that appropriate biosafety practices and procedures are rigorously followed and that the required physical containment features are maintained. If any element of biosafety is considered by the PI to be unachievable, the BSO should be notified immediately. A written report of the problem must be sent to the BSO within five working days. The responsibilities of the PI are more completely defined in Part I: Introduction.

b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction.

c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO.

d. Access to the laboratory is limited or restricted at the discretion of the PI when experiments are in progress.

a. Persons who may be at increased risk of acquiring infection or for whom infection might be unusually hazardous (e.g., immunocompromised persons) must be advised of their special risk in terms they understand, and must be discouraged from entering the laboratory.

b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 1 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research.

a. All persons who enter the BSL 1 laboratory must wear all required PPE. At a minimum, this includes coat or gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement.

b. Gloves should be worn if the skin on the hands is broken or if a rash is present. Alternatives to powdered latex gloves should be available.

c. All procedures are to be performed carefully to minimize the creation of splashes or aerosols. Protective eyewear should be worn for conduct of procedures in which splashes of microorganisms or other hazardous materials is anticipated.

d. No person shall leave the BSL 1 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility.

a. All cultures, stocks and other regulated wastes from a BSL 1 facility must be autoclaved before disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Materials to be decontaminated off-site must be packaged in accordance with applicable local, state and federal regulations before removal from the facility. Each laboratory must post clear instructions on how BSL 1 waste will be handled.

a. Each person will decontaminate his or her own work surfaces (e.g., benches, sinks, doors, handles, etc.) upon completion of work or at the end of the day and after any contamination with viable material.

b. Some disinfectants are much more effective than others and some organisms readily survive some disinfectants. The manufacturer or vendor can provide documentation of a disinfectant’s effectiveness against specific organisms.

c. Disinfectants are most effective when used according to the manufacturer’s instructions on concentration and contact time. Be sure to keep records that prove the liquid disinfectants are effective. Your laboratory-specific practices and procedures must include detailed instructions on the type of disinfectant, proper mix proportions, contact time required and other information that will ensure effective decontamination of work surfaces. EHS strongly discourages the use of flammable materials or disinfectants which may create vapors that will have toxic or adverse effects on laboratory staff. Bleach and industrial-grade Lysol are effective for most situations.

a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, notify Physical Plant and contact the BSO to help determine appropriate action.

a. Medical surveillance must be provided when appropriate.

a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory.

b. The BSO must be notified immediately if any significant problems, violations of biosafety practice, releases, spills or other laboratory accidents with potential biohazard exposure occur. If there is any doubt, the BSO should be notified.

All BSL 2 work practices and procedures must comply with the containment standards set forth in the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (4th Ed., 1999).

Each laboratory conducting BSL 2 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory.

The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 2 agents and those who work in or enter a BSL 2 facility. The practices and procedures set forth here are intended to augment the BSL 2 and ABSL 2 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment.

b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction.

c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO.

d. The PI must monitor and authorize access of all persons entering the BSL 2 laboratory. Access is limited to those who understand the nature of the biohazard, have adequate laboratory-specific biosafety training and agree to comply with all precautions. Visitors and maintenance personnel who enter the BSL 2 laboratory must be fully informed of the potential risks, required practices and procedures that they must follow. They must be instructed about the signs and symptoms of exposure to any and all biohazardous materials manipulated or stored in the laboratory.

b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 2 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research.

c. Cultures, tissues, etc., sent from a BSL 2 laboratory to another laboratory shall be handled using BSL 2 biosafety practices. All researchers receiving these biohazardous materials must be notified in writing of the risks associated with these materials and of the need to handle the materials using BSL 2 practices and procedures. Documentation of biohazardous materials transfers must include the institution, PI, date sent, nature and amount of material transferred and the BSL required for this work. Refer to Part XVIII: Transportation and Transfer for more detail.

a. All persons who enter the BSL 2 laboratory must wear all required PPE. At a minimum, this includes coat or gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement.

b. Any time a frozen storage container (freezer, ultra cold, liquid nitrogen) is opened, eye and face protection (face shield or goggles and a surgical mask) should be worn by persons handling the material.

c. Whenever potentially infected animals or tissues are in the BSL 2 laboratory and not contained in BSCs, all persons must wear face protection for anticipated splashes or sprays of infectious or other hazardous materials.

d. No person shall leave the BSL 2 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility.

a. All cultures, stocks and other regulated wastes from a BSL 2 facility must be autoclaved before disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Materials to be decontaminated off-site must be packaged in accordance with applicable local, state and federal regulations before removal from the facility. Each laboratory must post clear instructions on how BSL 2 waste will be handled.

a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, work in the BSL 2 facility must be halted. Notify Physical Plant and contact the BSO to help determine appropriate action.

b. Repair of the facility or laboratory equipment that requires entry into the BSL 2 laboratory by someone other than the normal laboratory staff must be authorized by the BSO before work begins. For after hours emergencies, contact the BSO through Campus Police at 619-594-1991.

c. Any time the BSL 2 facility must be closed for maintenance or repair, a laboratory clearance inspection must be performed by the BSO before the work may commence. Once clearance is granted, no further work with biohazardous materials may be conducted until all maintenance and repair work is completed. A thorough inspection of the laboratory must be conducted by the PI or designee to ensure that the laboratory is functioning properly before work with biohazardous materials may recommence.

d. Routine maintenance that affects ventilation or affects containment provided by the facility or requires entry into the laboratory by non-laboratory staff must be scheduled with EHS at least two weeks in advance. No maintenance or repair work may begin without prior EHS authorization. It is expected that EHS will conduct unannounced inspections during the maintenance.

e. Weekly tests of the required negative pressure for each room in the BSL 2 facility must be performed. (This test can be performed using a strip of tissue held in the opening of a door held slightly ajar). Problems must be reported without delay to Physical Plant and to the BSO. All results must be logged and kept on file.

f. Special containment systems such as an exhaust HEPA filtration system must be tested and certified to meet National Sanitation Foundation standard 49 no less than annually. These tests must be performed by an approved certification company.

g. The PI must keep a log of all maintenance conducted by non-laboratory staff when the BSL 2 facility has NOT been closed for maintenance. The log must record:

a. Medical surveillance must be provided for all persons who perform BSL 2 work. As appropriate, each person must be offered serologic testing when they begin work. Additional serum specimens may be collected periodically, depending on the agents handled or the function of the facility. The results of this serologic screening must be made available to the IBC upon request. Results may be coded to protect the privacy of the individuals. The IBC may use the results of these tests as a tool to evaluate the effectiveness of the laboratory’s biosafety program.

a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory.

Each laboratory conducting BSL 3 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory.

The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 3 agents and those who work in or enter a BSL 3 facility. The practices and procedures set forth here are intended to augment the BSL 3 and ABSL 3 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment.

b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction.

c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO.

d. The PI must monitor and authorize access of all persons entering the BSL 3 laboratory. Access is limited to those who understand the nature of the biohazard, have adequate laboratory-specific biosafety training and agree to comply with all precautions. Visitors and maintenance personnel who enter the BSL 3 laboratory must be fully informed of the potential risks, required practices and procedures that they must follow. They must be instructed about the signs and symptoms of exposure to any and all biohazardous materials manipulated or stored in the laboratory.

b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 3 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research.

c. Cultures, tissues, etc., sent from a BSL 3 laboratory to another laboratory shall be handled using BSL 3 biosafety practices. All researchers receiving these biohazardous materials must be notified in writing of the risks associated with these materials and of the need to handle the materials using BSL 3 practices and procedures. Documentation of biohazardous materials transfers must include the institution, PI, date sent, nature and amount of material transferred and the BSL required for this work. Refer to Part XVIII: Transportation and Transfer for more detail.

a. All persons who enter the BSL 3 laboratory must wear all required PPE. At a minimum, this includes wraparound gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement.

c. Whenever potentially infected animals or tissues are in the BSL 3 laboratory and not contained in BSCs, all persons must be provided with (and required to wear) a HEPA-filtered respirator. This respirator provides protection against both liquid and solid aerosols. The respirator must be NIOSH certified and fit tested by EHS.

d. No person shall leave the BSL 3 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility.

a. All cultures, stocks and other regulated wastes from a BSL 3 facility must be autoclaved before removal for off-site disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Each laboratory must post clear instructions on how BSL 3 waste will be handled.

a. Any item removed from the BSL 3 laboratory but not considered to be waste must be decontaminated before removal, even if there is only a small chance that it is contaminated. Items may be wiped down with a disinfectant or sealed in a bag and autoclaved.

b. Each person will decontaminate his or her own work surfaces (e.g., benches, sinks, doors, handles, etc.) immediately after each use and immediately after any contamination with viable material.

c. Some disinfectants are much more effective than others and some organisms readily survive some disinfectants. The manufacturer or vendor can provide documentation of a disinfectant’s effectiveness against specific organisms.

d. Disinfectants are most effective when used according to the manufacturer’s instructions on concentration and contact time. Be sure to keep records that prove the liquid disinfectants are effective. Your laboratory-specific practices and procedures must include detailed instructions on the type of disinfectant, proper mix proportions, contact time required and other information that will ensure effective decontamination of work surfaces. EHS strongly discourages the use of flammable materials or disinfectants which may create vapors that will have toxic or adverse effects on laboratory staff. Bleach and industrial-grade Lysol are effective for most situations.

a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, work in the BSL 3 facility must be halted. Notify Physical Plant and contact the BSO to help determine appropriate action.

b. Repair of the facility or laboratory equipment that requires entry into the BSL 3 laboratory by someone other than the normal laboratory staff must be authorized by the BSO before work begins. For after hours emergencies, contact the BSO through Campus Police at 619-594-1991.

c. Any time the BSL 3 facility must be closed for maintenance or repair, a laboratory clearance inspection must be performed by the BSO before the work may commence. Once clearance is granted, no further work with biohazardous materials may be conducted until all maintenance and repair work is completed. A thorough inspection of the laboratory must be conducted by the PI or designee to ensure that the laboratory is functioning properly before work with biohazardous materials may recommence.

e. Weekly tests of the required negative pressure for each room in the BSL 3 facility must be performed. (This test can be performed using a strip of tissue held in the opening of a door held slightly ajar). Problems must be reported without delay to Physical Plant and to the BSO. All results must be logged and kept on file.

g. The PI must keep a log of all maintenance conducted by non-laboratory staff when the BSL 3 facility has NOT been closed for maintenance. The log must record: Type of work/maintenance completed

a. Medical surveillance must be provided for all persons who perform BSL 3 work. As appropriate, each person must be offered serologic testing when they begin work. Additional serum specimens may be collected periodically, depending on the agents handled or the function of the facility. The results of this serologic screening must be made available to the IBC upon request. Results may be coded to protect the privacy of the individuals. The IBC may use the results of these tests as a tool to evaluate the effectiveness of the laboratory’s biosafety program.

a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory.

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	Environmental Health and Safety SDSU Biosafety Manual San Diego State University BIOHAZARD CONTROL PROGRAM Part IV: BIOSAFETY LEVEL PRACTICES PART IV. A. BIOSAFETY LEVEL PRACTICES CHART PART IV. B. BIOSAFETY LEVEL 1 PRACTICES PART IV. C. BIOSAFETY LEVEL 2 PRACTICES PART IV. D. BIOSAFETY LEVEL 3 PRACTICES A. BIOSAFETY LEVEL PRACTICES CHART B. BIOSAFETY LEVEL 1 PRACTICES All BSL 1 work practices and procedures must comply with the containment standards set forth in the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (4th Ed., 1999). Each laboratory conducting BSL 1 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory. 1. General Principles BSL 1 work, by definition, involves agents of no known hazard or of low potential hazard to personnel and the environment. BSL 1 practices, safety equipment and facilities are appropriate for most undergraduate training and teaching laboratories, and for other facilities in which work is done with defined and characterized strains of viable microorganisms not known to cause disease in healthy adult humans. BSL 1 represents a basic level of containment that relies on standard microbiological practices with no special primary or secondary barriers recommended, other than a sink for handwashing. The laboratory is not separated from the general traffic patterns in the building. Work is generally conducted on open bench tops. Special containment devices or equipment such as a biological safety cabinet are generally not required for manipulations of agents assigned to BSL 1. Laboratory personnel have specific training in the procedures conducted in the laboratory and are supervised by a scientist with general training in microbiology or related science. Standard Microbiological Practices a. Persons wash their hands after they handle viable materials, after removing gloves and before leaving the laboratory. b. Eating, drinking, smoking, handling contact lenses, applying cosmetics and storing food for human use are not permitted in the work areas. Persons who wear contact lenses in laboratories should also wear goggles or a face shield. Food is stored outside the work area in cabinets or refrigerators designated and used for this purpose only. c. Mechanical pipetting devices are used; pipetting by mouth is strictly prohibited. d. Policies for the safe handling of sharps are instituted. e. A biohazard sign can be posted at the entrance to the laboratory whenever infectious agents are present. The sign may include the name of the agent(s) in use and the name and telephone number of the investigator. f. An insect and rodent control program is in effect. (Example: If the laboratory has windows that open, they are fitted with fly screens). Laboratory Facilities a. The laboratory is designed so that it can be easily cleaned. Carpets and rugs in laboratories are not appropriate. b. Bench tops are impervious to water and resistant to acids, alkalis, organic solvents, moderate heat and chemicals used to decontaminate the work surface and equipment. c. Laboratory furniture is sturdy. Spaces between benches, cabinets and equipment are accessible for cleaning. 2. Specific Responsibilities, Practices and Procedures The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 1 agents and those who work in or enter a BSL 1 facility. The practices and procedures set forth here are intended to augment the BSL 1 and ABSL 1 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment. Responsibilities a. The PI must ensure that appropriate biosafety practices and procedures are rigorously followed and that the required physical containment features are maintained. If any element of biosafety is considered by the PI to be unachievable, the BSO should be notified immediately. A written report of the problem must be sent to the BSO within five working days. The responsibilities of the PI are more completely defined in Part I: Introduction. b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction. c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO. d. Access to the laboratory is limited or restricted at the discretion of the PI when experiments are in progress. Information a. Persons who may be at increased risk of acquiring infection or for whom infection might be unusually hazardous (e.g., immunocompromised persons) must be advised of their special risk in terms they understand, and must be discouraged from entering the laboratory. b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 1 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research. Personal Protective Equipment a. All persons who enter the BSL 1 laboratory must wear all required PPE. At a minimum, this includes coat or gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement. b. Gloves should be worn if the skin on the hands is broken or if a rash is present. Alternatives to powdered latex gloves should be available. c. All procedures are to be performed carefully to minimize the creation of splashes or aerosols. Protective eyewear should be worn for conduct of procedures in which splashes of microorganisms or other hazardous materials is anticipated. d. No person shall leave the BSL 1 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility. Biohazardous Material Waste a. All cultures, stocks and other regulated wastes from a BSL 1 facility must be autoclaved before disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Materials to be decontaminated off-site must be packaged in accordance with applicable local, state and federal regulations before removal from the facility. Each laboratory must post clear instructions on how BSL 1 waste will be handled. Decontamination a. Each person will decontaminate his or her own work surfaces (e.g., benches, sinks, doors, handles, etc.) upon completion of work or at the end of the day and after any contamination with viable material. b. Some disinfectants are much more effective than others and some organisms readily survive some disinfectants. The manufacturer or vendor can provide documentation of a disinfectant’s effectiveness against specific organisms. c. Disinfectants are most effective when used according to the manufacturer’s instructions on concentration and contact time. Be sure to keep records that prove the liquid disinfectants are effective. Your laboratory-specific practices and procedures must include detailed instructions on the type of disinfectant, proper mix proportions, contact time required and other information that will ensure effective decontamination of work surfaces. EHS strongly discourages the use of flammable materials or disinfectants which may create vapors that will have toxic or adverse effects on laboratory staff. Bleach and industrial-grade Lysol are effective for most situations. Laboratory Maintenance and Repair a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, notify Physical Plant and contact the BSO to help determine appropriate action. Medical Surveillance a. Medical surveillance must be provided when appropriate. Infections and Spills a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory. b. The BSO must be notified immediately if any significant problems, violations of biosafety practice, releases, spills or other laboratory accidents with potential biohazard exposure occur. If there is any doubt, the BSO should be notified. C. BIOSAFETY LEVEL 2 PRACTICES All BSL 2 work practices and procedures must comply with the containment standards set forth in the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (4th Ed., 1999). Each laboratory conducting BSL 2 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory. 1. General Principles BSL 2 work, by definition, involves agents of moderate potential hazard to personnel and the environment. The primary hazards to personnel working with these agents are autoinoculation, ingestion and inhalation of aerosols. 2. Specific Responsibilities, Practices and Procedures The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 2 agents and those who work in or enter a BSL 2 facility. The practices and procedures set forth here are intended to augment the BSL 2 and ABSL 2 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment. Responsibilities a. The PI must ensure that appropriate biosafety practices and procedures are rigorously followed and that the required physical containment features are maintained. If any element of biosafety is considered by the PI to be unachievable, the BSO should be notified immediately. A written report of the problem must be sent to the BSO within five working days. The responsibilities of the PI are more completely defined in Part I: Introduction. b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction. c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO. d. The PI must monitor and authorize access of all persons entering the BSL 2 laboratory. Access is limited to those who understand the nature of the biohazard, have adequate laboratory-specific biosafety training and agree to comply with all precautions. Visitors and maintenance personnel who enter the BSL 2 laboratory must be fully informed of the potential risks, required practices and procedures that they must follow. They must be instructed about the signs and symptoms of exposure to any and all biohazardous materials manipulated or stored in the laboratory. Information a. Persons who may be at increased risk of acquiring infection or for whom infection might be unusually hazardous (e.g., immunocompromised persons) must be advised of their special risk in terms they understand, and must be strongly discouraged from entering the laboratory. b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 2 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research. c. Cultures, tissues, etc., sent from a BSL 2 laboratory to another laboratory shall be handled using BSL 2 biosafety practices. All researchers receiving these biohazardous materials must be notified in writing of the risks associated with these materials and of the need to handle the materials using BSL 2 practices and procedures. Documentation of biohazardous materials transfers must include the institution, PI, date sent, nature and amount of material transferred and the BSL required for this work. Refer to Part XVIII: Transportation and Transfer for more detail. Personal Protective Equipment a. All persons who enter the BSL 2 laboratory must wear all required PPE. At a minimum, this includes coat or gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement. b. Any time a frozen storage container (freezer, ultra cold, liquid nitrogen) is opened, eye and face protection (face shield or goggles and a surgical mask) should be worn by persons handling the material. c. Whenever potentially infected animals or tissues are in the BSL 2 laboratory and not contained in BSCs, all persons must wear face protection for anticipated splashes or sprays of infectious or other hazardous materials. d. No person shall leave the BSL 2 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility. Biohazardous Material Waste a. All cultures, stocks and other regulated wastes from a BSL 2 facility must be autoclaved before disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Materials to be decontaminated off-site must be packaged in accordance with applicable local, state and federal regulations before removal from the facility. Each laboratory must post clear instructions on how BSL 2 waste will be handled. Decontamination a. Each person will decontaminate his or her own work surfaces (e.g., benches, sinks, doors, handles, etc.) upon completion of work or at the end of the day and after any contamination with viable material. b. Some disinfectants are much more effective than others and some organisms readily survive some disinfectants. The manufacturer or vendor can provide documentation of a disinfectant’s effectiveness against specific organisms. c. Disinfectants are most effective when used according to the manufacturer’s instructions on concentration and contact time. Be sure to keep records that prove the liquid disinfectants are effective. Your laboratory-specific practices and procedures must include detailed instructions on the type of disinfectant, proper mix proportions, contact time required and other information that will ensure effective decontamination of work surfaces. EHS strongly discourages the use of flammable materials or disinfectants which may create vapors that will have toxic or adverse effects on laboratory staff. Bleach and industrial-grade Lysol are effective for most situations. Laboratory Maintenance and Repair a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, work in the BSL 2 facility must be halted. Notify Physical Plant and contact the BSO to help determine appropriate action. b. Repair of the facility or laboratory equipment that requires entry into the BSL 2 laboratory by someone other than the normal laboratory staff must be authorized by the BSO before work begins. For after hours emergencies, contact the BSO through Campus Police at 619-594-1991. c. Any time the BSL 2 facility must be closed for maintenance or repair, a laboratory clearance inspection must be performed by the BSO before the work may commence. Once clearance is granted, no further work with biohazardous materials may be conducted until all maintenance and repair work is completed. A thorough inspection of the laboratory must be conducted by the PI or designee to ensure that the laboratory is functioning properly before work with biohazardous materials may recommence. d. Routine maintenance that affects ventilation or affects containment provided by the facility or requires entry into the laboratory by non-laboratory staff must be scheduled with EHS at least two weeks in advance. No maintenance or repair work may begin without prior EHS authorization. It is expected that EHS will conduct unannounced inspections during the maintenance. e. Weekly tests of the required negative pressure for each room in the BSL 2 facility must be performed. (This test can be performed using a strip of tissue held in the opening of a door held slightly ajar). Problems must be reported without delay to Physical Plant and to the BSO. All results must be logged and kept on file. f. Special containment systems such as an exhaust HEPA filtration system must be tested and certified to meet National Sanitation Foundation standard 49 no less than annually. These tests must be performed by an approved certification company. g. The PI must keep a log of all maintenance conducted by non-laboratory staff when the BSL 2 facility has NOT been closed for maintenance. The log must record: Type of work/maintenance completed Date of entry Name of workers Start time Completion time Work order number Medical Surveillance a. Medical surveillance must be provided for all persons who perform BSL 2 work. As appropriate, each person must be offered serologic testing when they begin work. Additional serum specimens may be collected periodically, depending on the agents handled or the function of the facility. The results of this serologic screening must be made available to the IBC upon request. Results may be coded to protect the privacy of the individuals. The IBC may use the results of these tests as a tool to evaluate the effectiveness of the laboratory’s biosafety program. Infections and Spills a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory. b. The BSO must be notified immediately if any significant problems, violations of biosafety practice, releases, spills or other laboratory accidents with potential biohazard exposure occur. If there is any doubt, the BSO should be notified. D. BIOSAFETY LEVEL 3 PRACTICES All BSL 3 work practices and procedures must comply with the containment standards set forth in the CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories (4th Ed., 1999). Each laboratory conducting BSL 3 work must have written laboratory-specific biosafety practices and procedures approved by the IBC prior to commencement of work. It is highly recommended that the practices and procedures set forth here be adopted into the laboratory-specific practices and procedures and implemented as part of the required performance of every individual working in the laboratory. 1. General Principles BSL 3 work, by definition, involves agents that may cause serious and potentially lethal infection. The primary hazards to personnel working with these agents are autoinoculation, ingestion and inhalation of aerosols. 2. Specific Responsibilities, Practices and Procedures The following is provided to give guidance in the preparation of laboratory-specific biosafety practices and procedures. This part is intended to clarify the responsibilities of the PI, all persons who work with BSL 3 agents and those who work in or enter a BSL 3 facility. The practices and procedures set forth here are intended to augment the BSL 3 and ABSL 3 practices and procedures found in the CDC/NIH publication Biosafety in Microbiological and Biological Laboratories (4th Ed., 1999) and are not intended to limit or reduce those standards of containment. Responsibilities a. The PI must ensure that appropriate biosafety practices and procedures are rigorously followed and that the required physical containment features are maintained. If any element of biosafety is considered by the PI to be unachievable, the BSO should be notified immediately. A written report of the problem must be sent to the BSO within five working days. The responsibilities of the PI are more completely defined in Part I: Introduction. b. All persons must conscientiously follow the laboratory-specific biosafety practices and procedures. Responsibilities of laboratory staff are more completely defined in Part I: Introduction. c. Workers should observe and monitor the work practices and procedures of others in the laboratory. A serious or consistent failure to follow these guidelines must be reported to the PI or the BSO. d. The PI must monitor and authorize access of all persons entering the BSL 3 laboratory. Access is limited to those who understand the nature of the biohazard, have adequate laboratory-specific biosafety training and agree to comply with all precautions. Visitors and maintenance personnel who enter the BSL 3 laboratory must be fully informed of the potential risks, required practices and procedures that they must follow. They must be instructed about the signs and symptoms of exposure to any and all biohazardous materials manipulated or stored in the laboratory. Information a. Persons who may be at increased risk of acquiring infection or for whom infection might be unusually hazardous (e.g., immunocompromised persons) must be advised of their special risk in terms they understand, and must be strongly discouraged from entering the laboratory. b. Biosafety training must be regularly scheduled and presented to all persons who work in or who enter the BSL 3 laboratory. Training must focus on biosafety or other health and safety policies, practices and procedures to be followed for this research. c. Cultures, tissues, etc., sent from a BSL 3 laboratory to another laboratory shall be handled using BSL 3 biosafety practices. All researchers receiving these biohazardous materials must be notified in writing of the risks associated with these materials and of the need to handle the materials using BSL 3 practices and procedures. Documentation of biohazardous materials transfers must include the institution, PI, date sent, nature and amount of material transferred and the BSL required for this work. Refer to Part XVIII: Transportation and Transfer for more detail. Personal Protective Equipment a. All persons who enter the BSL 3 laboratory must wear all required PPE. At a minimum, this includes wraparound gown, gloves and eye protection. Shoe covers, masks, head covers, sleeve protectors and face shields should be added as appropriate. The PI and laboratory staff must enforce this requirement. b. Any time a frozen storage container (freezer, ultra cold, liquid nitrogen) is opened, eye and face protection (face shield or goggles and a surgical mask) should be worn by persons handling the material. c. Whenever potentially infected animals or tissues are in the BSL 3 laboratory and not contained in BSCs, all persons must be provided with (and required to wear) a HEPA-filtered respirator. This respirator provides protection against both liquid and solid aerosols. The respirator must be NIOSH certified and fit tested by EHS. d. No person shall leave the BSL 3 laboratory while wearing clothing designated for wear inside the laboratory. Distinctive protective clothing (clearly different from protective clothing used in nearby areas) shall be provided to the laboratory staff for their use inside the facility. Biohazardous Material Waste a. All cultures, stocks and other regulated wastes from a BSL 3 facility must be autoclaved before removal for off-site disposal. Materials to be decontaminated outside of the immediate laboratory are placed in a durable, leakproof container and closed for transport from the laboratory. Each laboratory must post clear instructions on how BSL 3 waste will be handled. Decontamination a. Any item removed from the BSL 3 laboratory but not considered to be waste must be decontaminated before removal, even if there is only a small chance that it is contaminated. Items may be wiped down with a disinfectant or sealed in a bag and autoclaved. b. Each person will decontaminate his or her own work surfaces (e.g., benches, sinks, doors, handles, etc.) immediately after each use and immediately after any contamination with viable material. c. Some disinfectants are much more effective than others and some organisms readily survive some disinfectants. The manufacturer or vendor can provide documentation of a disinfectant’s effectiveness against specific organisms. d. Disinfectants are most effective when used according to the manufacturer’s instructions on concentration and contact time. Be sure to keep records that prove the liquid disinfectants are effective. Your laboratory-specific practices and procedures must include detailed instructions on the type of disinfectant, proper mix proportions, contact time required and other information that will ensure effective decontamination of work surfaces. EHS strongly discourages the use of flammable materials or disinfectants which may create vapors that will have toxic or adverse effects on laboratory staff. Bleach and industrial-grade Lysol are effective for most situations. Laboratory Maintenance and Repair a. The PI is responsible for ensuring that the physical components of the laboratory designed to contain the biohazards associated with the research are working properly and are properly maintained (e.g., ventilation, filtration, sanitation, security). If the ventilation system or other physical containment component of the laboratory fails, work in the BSL 3 facility must be halted. Notify Physical Plant and contact the BSO to help determine appropriate action. b. Repair of the facility or laboratory equipment that requires entry into the BSL 3 laboratory by someone other than the normal laboratory staff must be authorized by the BSO before work begins. For after hours emergencies, contact the BSO through Campus Police at 619-594-1991. c. Any time the BSL 3 facility must be closed for maintenance or repair, a laboratory clearance inspection must be performed by the BSO before the work may commence. Once clearance is granted, no further work with biohazardous materials may be conducted until all maintenance and repair work is completed. A thorough inspection of the laboratory must be conducted by the PI or designee to ensure that the laboratory is functioning properly before work with biohazardous materials may recommence. d. Routine maintenance that affects ventilation or affects containment provided by the facility or requires entry into the laboratory by non-laboratory staff must be scheduled with EHS at least two weeks in advance. No maintenance or repair work may begin without prior EHS authorization. It is expected that EHS will conduct unannounced inspections during the maintenance. e. Weekly tests of the required negative pressure for each room in the BSL 3 facility must be performed. (This test can be performed using a strip of tissue held in the opening of a door held slightly ajar). Problems must be reported without delay to Physical Plant and to the BSO. All results must be logged and kept on file. f. Special containment systems such as an exhaust HEPA filtration system must be tested and certified to meet National Sanitation Foundation standard 49 no less than annually. These tests must be performed by an approved certification company. g. The PI must keep a log of all maintenance conducted by non-laboratory staff when the BSL 3 facility has NOT been closed for maintenance. The log must record: Type of work/maintenance completed Date of entry Name of workers Start time Completion time Work order number Medical Surveillance a. Medical surveillance must be provided for all persons who perform BSL 3 work. As appropriate, each person must be offered serologic testing when they begin work. Additional serum specimens may be collected periodically, depending on the agents handled or the function of the facility. The results of this serologic screening must be made available to the IBC upon request. Results may be coded to protect the privacy of the individuals. The IBC may use the results of these tests as a tool to evaluate the effectiveness of the laboratory’s biosafety program. Infections and Spills a. The BSO must be notified immediately if a person is known to have or suspected of having acquired an infection as a result of work in or around the laboratory. b. The BSO must be notified immediately if any significant problems, violations of biosafety practice, releases, spills or other laboratory accidents with potential biohazard exposure occur. If there is any doubt, the BSO should be notified. Note: documents in Portable Document Format (PDF) require Adobe Acrobat Reader 5.0 or higher to view; download Adobe Acrobat Reader.
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Environmental Health and Safety

SDSU Biosafety Manual

Part IV: BIOSAFETY LEVEL PRACTICES

Part IV:
BIOSAFETY LEVEL PRACTICES