Abstract
The Guidelines for Biosafety Training Programs for Workers Assigned to BSL-3 Research Laboratories were developed by biosafety professionals who oversee training programs for the 2 national biocontainment laboratories (NBLs) and the 13 regional biocontainment laboratories (RBLs) that participate in the National Institute of Allergy and Infectious Diseases (NIAID) NBL/RBL Network. These guidelines provide a general training framework for biosafety level 3 (BSL-3) high-containment laboratories, identify key training concepts, and outline training methodologies designed to standardize base knowledge, understanding, and technical competence of laboratory personnel working in high-containment laboratories. Emphasis is placed on building a culture of risk assessment–based safety through competency training designed to enhance understanding and recognition of potential biological hazards as well as methods for controlling these hazards. These guidelines may be of value to other institutions and academic research laboratories that are developing biosafety training programs for BSL-3 research.
The guidelines that have been developed for workers in BSL-3 laboratories provide a training framework, identify training concepts, and outline methods designed to standardize base knowledge, understanding, and technical competence of lab personnel working in high-containment laboratories. Emphasis is placed on building a culture of risk assessment–based safety through competency training designed to enhance understanding and recognition of potential biological hazards and methods for controlling these hazards.
NIAID funded the construction of 2 national biocontainment laboratories (NBLs) and 13 regional biocontainment laboratories (RBLs). The NBLs house biosafety level (BSL) 2, 3, and 4 laboratories; the RBLs include BSL-2 and -3 laboratories. In these secure, state-of-the-art facilities, researchers pursue the development of vaccines to immunize the public against diseases caused by emerging infectious agents and bioterrorism agents, including those regulated as select agents; develop and refine diagnostic tests to help first responders and other medical personnel rapidly detect exposure and provide treatment; and develop and test therapies to help patients exposed to bioterrorism agents regain their health. NIAID also established 11 regional centers of excellence for biodefense and emerging infectious diseases (RCEs), located throughout the United States, which support research on countering threats from bioterror agents and emerging infectious diseases.
In addition to the US government's multifaceted efforts to advance our nation's defenses against bioterrorism, many private and public institutions nationwide are constructing new high-containment facilities 1 and funding biomedical research involving high-risk pathogens, outbreaks of which occur naturally, frequently, and globally. 2 In 2004, 415 BSL-3 laboratories were registered with the Centers for Disease Control and Prevention (CDC) for storage and/or use of select agents; by 2010, this number had grown to 1,495, about half of which were academic. 3 In 2005, the Laboratory Response Network, a group of laboratories prepared to respond to acts of terrorism, emerging diseases, and public health emergencies, contained 120 labs with BSL-3 facilities; by 2011, this number had grown to 150. 4 This rapid expansion in the number of high-containment laboratories has created an urgent need for scientists, laboratory technicians, operations and maintenance specialists, and laboratory support personnel who are thoroughly trained in the principles and practices of biosafety and who are highly competent to carry out and assist in the conduct of safe science.
It is critical to note that the recent expansion of high-containment facilities has not resulted in a matching expansion of laboratory accidents, thefts, injuries, or releases of dangerous pathogens. High-containment laboratories must register with the CDC and/or the US Department of Agriculture (USDA) for all storage and use of select agents, and they must gain approval for each individual who will have access to select agents. Once registered, institutions are required by law to notify CDC or USDA of any accidents and near-misses, particularly those that could lead to theft, loss, or release of the agents. These facilities have an excellent safety record. As of September 2009, nearly 400 institutions had registered with CDC or USDA to work with select agents, with a total of almost 14,000 laboratory personnel approved for access to select agents. 5 Within a year prior to this publication, there had been a total of 7 laboratory-acquired infections of select agents reported to CDC, and at the time of this publication, there have been no reported thefts of select agents out of registered facilities and no reported infections of select agents in the public that have resulted from laboratory accidents. 6
The recent expansion of high-containment facilities has led leaders in the field to recognize the need to consistently and thoroughly train its scientists, laboratory technicians, and support personnel. Leaders of the majority of the operating BSL-4 facilities in the United States and Canada have agreed on a framework for training. 7 Direct components of training—such as the use of safety equipment like the Class III biological safety cabinet, use of personal protective equipment like the protective suit, and decontamination of infectious wastes—were identified as critical components for training personnel and for safe operation of BSL-4 facilities. A broad range of training methodologies was suggested, beginning with classroom training, moving to hands-on training, and concluding with one-on-one mentoring. Indirect components of training were identified as equally critical—for example, strong institutional support of the facility director in conjunction with the biosafety officer and operations manager to enforce safety standards and properly operate the facility. It is hoped that this article will provide a matching framework for training at BSL-3.
Emphasis on Competence
The Biosafety in Microbiological and Biomedical Laboratories (BMBL) 8 guidelines emphasize the importance of risk assessment and the technical competence of laboratory workers in establishing and sustaining a safe containment environment. Research programs requiring BSL-3 microbiological practices and containment are likely to involve indigenous or exotic agents that may cause serious or potentially lethal disease through the inhalation route of exposure. Laboratory scientists and support staff who are being trained to work in BSL-3 laboratories and animal biosafety level 3 (ABSL-3) facilities need to be able to assess the risks associated with handling BSL-3 agents in the laboratory and learn how to mitigate those risks identified to keep themselves, their colleagues, and the environment safe. To do this, laboratory scientists and support staff need to acquire skills so that they are proficient in standard and special microbiological practices and in good microbiological technique. These individuals must also be familiar with and be instructed on proper use of containment equipment and personal protective equipment. As expected, workers with previous laboratory and animal experience at BSL-1 and BSL-2 often demonstrate competence and proficiency at BSL-3 in a timelier manner when compared to others without this experience. The critical concepts and work practices include, but are not limited to, understanding of how infectious agents are transmitted in the research setting, familiarity with biological hazards and their controls, proper use of biological safety cabinets, and safe handling of research animals. A laboratory worker who is well-trained and proficient in these areas will minimize the numerous risks associated with infectious disease research. Approval of a trainee to work independently at BSL-3 or ABSL-3 is the responsibility of the laboratory director or principal investigator, following consultation with the trainee's mentor. This decision should be based on the trainee's ability to demonstrate that he or she has acquired the technical proficiency necessary to conduct safe practices associated with the laboratory's operational and research standard operating procedures for handling high-risk pathogens, and an assessment of the trainee's knowledge of the disease risks of the agents handled in the laboratory. This level of technical expertise and understanding will protect the laboratory worker, his or her colleagues, and the environment from the hazards associated with the research program, and it will promote a greater awareness of the importance of laboratory safety in high-containment laboratories. Understanding the principles of biosafety and adherence to the microbiological practices, containment, and facility safeguards described in the BMBL will contribute to a safer and healthier work environment for laboratory staff and adjacent personnel and for the community.
Competency guidelines for laboratory biosafety have been the focus of 2 recent federal reports that highlight the principle that competency training is critical for achieving the necessary expertise to work safely in the BSL-3 and ABSL-3 environment. Report of the Trans-Federal Task Force on Optimizing Biosafety and Biocontainment Oversight noted, “It is important to ensure that all personnel who work in, oversee, support, or manage high or maximum containment research laboratories achieve and maintain a sufficient level of technical competence for maintaining safety in those facilities.”9(p94) CDC and the Association of Public Health Laboratories' (APHL) “Guidelines for Biosafety Laboratory Competency” set forth competency guidelines “intended for laboratorians working with hazardous biologic agents, obtained from either samples or specimens that are maintained and manipulated in clinical, environmental, public health, academic, and research laboratories.”10(p1) While these federal documents provide general guidelines for training and biosafety oversight, the practical application and recommendations for content of this training were not specified. This article attempts to provide methodical guidelines to achieve this level of competence for individuals working at BSL-3.
Key Training Concepts
All personnel who work in, support, manage, or provide oversight to high-containment laboratories should receive training in 10 key concepts: (1) fundamental principles of biosafety and biocontainment; (2) specific information about the hazardous characteristics of each agent and toxin used in the facility; (3) proper care and use of personal protective equipment; (4) proper care and use of laboratory safety equipment; (5) medical surveillance; (6) animal biosafety, when applicable; (7) relevant federal, state, and local regulations; (8) institutional and facility policies; (9) facility-specific operating procedures; and (10) biosecurity. Training for each element should include both universal information common to all facilities and specific information based on the design and operation of the individual facility. In addition, training should be role-specific and based on the individual needs and experience of personnel.
1. Fundamental Principles of Biosafety and Biocontainment
The purpose of biocontainment is to protect laboratory workers, others in the work area, and the outside environment from exposure to potentially infectious materials. 8 Trainees should receive thorough instruction on the key elements of containment, which include laboratory practices and techniques, use of safety equipment, and proper facility design and construction. Each individual should receive training that addresses information pertinent to his or her specific duties and responsibilities.
2. Hazardous Characteristics of Each Agent and Toxin
A biosafety training program should provide training on the hazards associated with each agent or toxin used in the facility so that personnel are aware of the risks in handling the agent or toxin, the diseases or illnesses that an agent or toxin can cause, signs and symptoms of disease, modes of transmission and possible routes of exposure in the research setting, procedures to follow in the event of a known or suspected exposure, and reporting procedures. All research and support personnel should receive this training.
3. Proper Care and Use of Personal Protective Equipment
All personnel must receive training on the personal protective equipment (PPE) that they will be required to use when they enter the BSL-3 or ABSL-3 laboratory. The training should address the level of protection provided; medical surveillance requirements, such as a pulmonary function test and/or respiratory fit testing; how to check the integrity of PPE; how to properly put on and wear protective laboratory clothing, gloves, and face, eye, and respiratory PPE; what to do if PPE fails or if one experiences an allergic reaction or other complication associated with PPE; proper disposal of PPE; and care and maintenance of reusable PPE.
4. Proper Care and Use of Laboratory Safety Equipment
Personnel must receive training in the proper use, care, maintenance, certification, and validation of containment and laboratory equipment. Training should enable personnel to attain a level of proficiency in the proper operation and use of biological safety cabinets, which serve as the primary means of containing aerosols generated from manipulation of infectious agents. Training should also enable proficiency in the operation of other engineering controls that are likely to be present in the BSL-3 or ABSL-3 laboratory, such as centrifuge safety rotors and safety lids and necropsy tables.
Personnel must also be aware of the potential risks associated with each type of engineering control and be able to implement safeguards to minimize risk. For example, certain laboratory equipment may have the potential to generate aerosols (eg, flow cytometers, bone saws, centrifuges); therefore, trainees should be educated on how to recognize and minimize the formation of aerosols. Training should be provided on equipment used to inactivate infectious materials, such as steam sterilizers. Training should focus on proper use and care of the equipment, certification and validation, facility or institutional procedures to follow in the event of an equipment failure or malfunction, and routine maintenance requirements for the equipment.
5. Medical Surveillance
Each individual must receive information and training on medical surveillance requirements relevant to his or her position and duties, including entry requirements, periodic health evaluations, vaccination requirements, respirator fit testing, baseline and/or periodic serum banking if required, and serum surveillance. The training should include instruction on the proper response and notification procedures in the event of a known or suspected exposure or the onset of symptoms that could be due to an inadvertent exposure.
6. Animal Biosafety
ABSL-3 involves practices suitable for work with laboratory animals infected with indigenous or exotic agents, agents that present a potential for aerosol transmission, and agents causing serious or potentially lethal disease. Because of the high risk of transmission through inhalation of aerosols, ingestion, or cutaneous or mucous membrane exposure, personnel must undergo agent-specific training to understand the potential hazards of working with infected animals, such as bites, scratches, needlesticks, or scalpel cuts. Before training at ABSL-3, personnel should demonstrate proficiency at ABSL-2 in animal handling and husbandry procedures. ABSL-3 training should include a rigorous hands-on plan for a minimum period of time to allow personnel to gain experience with each animal species. The training requirements for species may vary, depending on the comfort level and skill of the individual. Additional training may include use of a chemical or physical restraint, use of supplemental PPE, use and disposal of safety-engineered sharps, necropsy tables, animal containment cages, cage changing stations, loose-animal or escaped-animal protocols, animal waste decontamination and disposal, and animal carcass disposal. All work with laboratory animals should be conducted with care to ensure both the safety of the laboratory worker and the welfare of the animal.
7. Relevant Federal, State, and Local Regulations
Compliance with regulatory requirements and guidelines regarding BSL-3 and ABSL-3 facilities is the shared responsibility of all personnel who work in, support, manage, or provide oversight for the facility. Personnel should be aware of regulations governing work at the facility, particularly the work in which they are involved, and policies and procedures in place to comply with the regulations. Federal regulations and guidelines include the NIH Guidelines for Research Involving Recombinant DNA Molecules (“NIH Guidelines”); Biosafety in Microbiological and Biomedical Laboratories (BMBL); Title 29, Labor, CFR Part 1910.1030 (Bloodborne Pathogens standard); Occupational Safety and Health Act of 1970, Section 5 (General Duty Clause); and Code of Federal Regulations Parts 73, 121, and 331 (Select Agents). There may be applicable state and local guidelines that apply to each institution as well.
8. Institutional and Facility Policies
All personnel must receive training in institutional and facility policies regarding security and access, visitor access, entry requirements, hazardous waste disposal, the use of sharps, the review and approval of research protocols prior to the start of work, emergency response, and any additional facility-specific policies.
9. Standard Operating Procedures
Personnel should be trained on laboratory-specific standard operating procedures (SOPs). These might include entry and exit procedures, waste management, decontamination and disinfection, management of sharps, shipping and transport of infectious material, inactivation of pathogens, security procedures (eg, access restrictions, suspicious packages, suspicious people, bomb threats, theft, loss or release of material), incident response procedures (eg, spill response, emergency evacuation, extreme weather, power outage, air flow disruption, failure of the communication system, medical emergency, exposure response), use of equipment, and experimental protocols. Training on SOPs should include supervised hands-on instruction that allows personnel to develop and master the skills necessary to safely carry out their work independently. All personnel should be encouraged to report any concerns or incidents to their supervisor and/or biosafety officer without fear of repercussion.
10. Biosecurity
Training in the fundamentals of biosecurity is increasingly important for personnel handling BSL-3 agents and select agents. Although many biosafety programs contain key elements of biosecurity, the importance of biosecurity measures should be emphasized in training. Biosecurity training should include methods at the institution to protect against theft, loss, or misuse of BSL-3 agents and select agents. At a minimum, the primary methods of biosecurity should be discussed, such as physical security elements for facility, laboratory, storage, and agent access controls (eg, perimeter fences, entrances and exits, security cameras, identification badges, key locks, proximity card readers, paper log books); protection of sensitive information technology (IT) (eg, handling, use and storage of select agent inventory data and laboratory notebooks); material accountability (eg, inventory and material transfers); and personal reliability (eg, reporting unknown or suspicious people in the restricted-access facility, and the visitor policy). Enhanced biosecurity training might include recognition of and reporting of concerning behaviors among peers that could potentially compromise the security of infectious agents, and understanding of institutional protocols to evaluate research protocols for potential dual-use applications.
Training Framework
The training framework provides a methodology for conducting the training program. The training framework establishes a program that includes a combination of: (1) didactic classroom training on the key BSL-3/ABSL-3 elements described above; (2) supervised practical training and exercises for learning the standard operating procedures of a working BSL-3/ABSL-3 laboratory; (3) mentorship training for learning BSL-3/ABSL-3 safe practices used in the conduct of research projects; and (4) annual refresher training.
Didactic Classroom Training
The didactic classroom training course addresses the fundamental principles of biosafety and biocontainment; primary and secondary barriers that protect laboratory personnel, the community, and the environment; the primary hazards of agents that are handled in the BSL-3 laboratory; and the technical skill that is required to prevent exposures to those agents when handled in biological safety cabinets and other containment equipment. The didactic training course also introduces the trainees to the design and construction elements that provide secondary containment for the BSL-3/ABSL-3 laboratory, including the BSL-3 barriers; directional airflow; high-efficiency particulate air (HEPA) filtration of exhaust air; double-door pass-through autoclaves; and sealed seams, floors, walls, and ceiling surfaces of the laboratory space. Finally, the didactic classroom training may include animal biosafety topics such as animal handling, physical and chemical restraints, supplemental protective equipment, specialty animal equipment, and waste and carcass disposal, for trainees who will work with infected animals.
Supervised Practical Training and Exercises
The second phase of the training framework introduces the trainee to the standard operating procedures of the BSL-3/ABSL-3 and may include both didactic training and supervised practical training. This phase of the training should be specific to the institution and facility and may include access procedures for entering the BSL-3/ABSL-3, wearing of PPE, safety procedures and policies for the safe handling of sharps, decontamination methods for surfaces and waste materials, techniques for safely handling agents in biological safety cabinets, use of pipetting aids, safe work with infected animals, alarms and emergency operations, spill clean-up procedures, and other practices and procedures. The second phase of training should be led by an individual with appropriate knowledge and experience. The biological safety officer and the facility operations manager often participate in the second-phase training program.
Mentorship Training
The third training phase provides mentorship training in the BSL-3/ABSL-3. This training begins after the trainee has successfully completed phases 1 and 2 of the training program. Training should proceed at a pace that is comfortable and appropriate for the trainee. Trainees should not progress to the next set of tasks or training phase until they have demonstrated proficiency in their present set of tasks. Mentorship training should also account for different levels of experience and expertise. Refer to Conducting the Training Plan (below) for more details on mentored training.
Annual Refresher Training
Annual refresher training is required to maintain the skills and knowledge acquired during earlier training sessions and is important in promoting a safety culture in the BSL-3/ABSL-3 environment. Annual refresher training is also required by the BMBL, the OSHA bloodborne pathogens standard, the NIH Guidelines, and the select agent regulations.
Conducting the Training Plan
The initial phase of the BSL-3/ABSL-3 training (didactic classroom training) should begin only after a trainee has demonstrated proficiency in a BSL-2 and an ABSL-2 facility if animal research will be conducted. As stated above, the initial phase of the training includes fundamental information pertaining to the operation and policies of a BSL-3/ABSL-3—for example, facility design and construction, principles of biosafety, disinfection and decontamination, use of a biological safety cabinet, and other pertinent information for working safely in containment. The second phase of the training (supervised practical training and exercises) should follow and expand on the principles explained in phase 1, with a focus on the institution's and facility's design and operation, safety practices, entry and exit procedures, and the like.
Both the initial phase of training and the second phase of training can be presented in a variety of ways, depending on institution-specific factors such as class size, composition of the training team, and training resources. For example, if resources allow for small class sizes, trainees may feel more engaged with the trainer and may be encouraged to ask more questions and to take a more active role in their training, as compared to a larger class. If resources are available for the development of online training modules, this may enable the training of personnel with competing job duties that might preclude participation in full-day or multiday training. Use of tools such as homework and classroom exercises may allow certain components of the training to be emphasized. The order of training sessions may be tailored to fit the needs of trainees; in some circumstances, there may be a requirement to complete didactic training sessions in a specific order so that there is a sequential development of understanding of concepts. In other circumstances, a person might begin his or her training at any stage. Training laboratories may prove especially useful during the second phase of training so that trainees can practice donning and doffing protective equipment and using safety equipment such as biological safety cabinets.
The second phase of training may provide opportunities to incorporate adult learning techniques into the training program. These techniques may include emphasizing the personal benefits of training to the trainee by directly connecting the training outcomes to each employee's position and responsibilities, encouraging participation and sharing of professional experiences, and using a variety of teaching methods with emphasis on active learning techniques such as case studies, role playing, and games. The second phase of training may also provide opportunities to emphasize the importance of following the training materials and associated standard procedures as well as the potential ramifications of failure to comply with acceptable procedures.
Once the trainee completes phases 1 and 2 in the basics of working in a BSL-3 or ABSL-3 facility, mentored training begins. A proper mentor must have extensive experience and expertise in the specific work tasks assigned to the new trainee. Qualified mentors for research personnel are often senior laboratory technicians, postdoctoral fellows, or principal investigators. Support personnel, such as maintenance mechanics, are best mentored by experienced experts in their occupational discipline. An experienced mentor with the proper expertise in the assigned task may not always be immediately available on-site. This is especially problematic for new high-containment laboratories. In such cases, it may be necessary to reach out to experienced workers at other high-containment facilities and seek their help with a mentoring plan. The plan may include off-site visits to other active containment labs, and it could also include hosting an experienced expert at your facility for the purpose of starting the mentoring process. The mentor will work with the trainee to provide hands-on training, beginning with a tour and orientation of the laboratory. Hands-on training focuses on the facility's approved SOPs, which cover basic safety procedures, day-to-day operations, each experimental protocol and procedure performed in the laboratory, and possible emergencies. Many facilities have a minimum hourly requirement for the hands-on training portion that includes 2 parts: part 1 in which the trainee observes the experienced worker, and part 2 in which the trainee performs work under the guidance of the experienced worker. Mentored training must include animal training before the trainee handles infected animals. When the mentor deems the trainee competent in the appropriate SOPs, and the trainee feels confident, the trainee can complete any final clearance requirements and become a fully approved BSL-3/ABSL-3 user. Typically, this final step is either in the form of a written quiz or direct observation by the facility manager and biosafety officer who will assess the safety competence of the trainee.
The success of the biosafety program at any given institution depends on the support of many different people. Personnel safety and prevention of exposures are of utmost importance; hence, it is vital for principal investigators to emphasize the importance of biosafety to their laboratory staff. The investigator should be the first in his or her laboratory to complete biosafety training at the institution, and he or she should be willing to undergo further training and observation if he or she is new to work at BSL-3, not familiar with the hazards associated with the project, or has been assigned to a new BSL-3 facility with different engineering controls and policies. The culture of biosafety is enhanced and is at its best when the principal investigators, laboratory workers, and biosafety officers work together to discuss issues and possible exposures and to implement new ideas to improve the training program.
Competence, experience, knowledge of the agent and procedure hazards, good habits, caution, attentiveness, and concern for the health of coworkers are prerequisites for a laboratory staff in order to reduce the inherent risks of work with hazardous agents. Not all workers who join a laboratory staff will have these traits, even though they may possess excellent credentials. Laboratory directors or principal investigators should train and retrain new staff to the point where aseptic techniques and safety precautions become second nature. 11
Competency Training Objectives
It is important that each course include training objectives. Training objectives set the course goals, help frame the course curriculum, and provide elements on which the trainee's competence and understanding can be measured. Appendix A, “Curricula Development for Didactic Biosafety Training Courses,” is an example of a didactic safety training course based on defined learning objectives (see http://online.liebertpub.com/loi/BSP). This type of course structure ensures consistency in course training and learning expectations. Appendix B includes an excerpt of the suggested competency guidelines as presented in the CDC/APHL “Guidelines for Biosafety Laboratory Competency” for entry-level, midlevel, and senior-level laboratory personnel working with biologic hazards (see http://online.liebertpub.com/loi/BSP). These guidelines would serve as an excellent source on which to develop competency courses and corresponding training objectives.
Assessing Competence
An individual's competence and ability to conduct work in a BSL-3 or ABSL-3 should be evaluated at each step of the training by all stakeholders, including the principal investigator, the facility director or biosafety officer, and the trainee. The training team should determine the minimum competencies for each trainee, based on the position and experience of the trainee as well as the anticipated tasks that the trainee will perform, and then the competency should be measured and documented to provide a written training record. The competency guidelines as presented in the CDC/APHL “Guidelines for Biosafety Laboratory Competency” may prove useful. Such competencies should be reassessed and revised when new procedures are added. For example, during didactic classroom training, a written or oral exam may be used to verify that the trainee understands concepts, SOPs, and emergency procedures. Exam results can be retained in the training record. The trainer can then assess whether the trainee is ready to move on to supervised practical training and exercises. During this phase, laboratory-based demonstrations of knowledge may be more suitable to verify that the trainee understands the concepts and procedures, although exams can be used as a supplement. Laboratory-based demonstrations can be documented and retained in the training record. The trainer can then assess whether the trainee is ready to move on to mentored training. During this phase, by virtue of the mentor's close oversight of the trainee, the mentor can assess whether the trainee has gained the appropriate level of proficiency to work independently in the facility. Checklists of specific tasks and skills can be useful to document proficiency during mentored training. At all times during the training process, the trainer should be able to consult with other stakeholders about the trainee's competence. Evaluation of the trainee's competence can continue after he or she is granted the ability to work independently in the BSL-3/ABSL-3 facility; for example, the trainee's safety record can be periodically assessed to assure continued adherence to established procedures.
The Importance of Risk Assessment
Biological risk assessment is a process used to identify the hazardous characteristics of a known infectious or potentially infectious agent or material, the practices 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. Directors and principal investigators of microbiological and biomedical laboratories have the primary responsibility to conduct a risk assessment for research projects that involve pathogens before approving implementation of the research project. They also have the responsibility to assess the overall risks of the work in the BSL-3 or ABSL-3, typically categorized as agent hazards and laboratory procedure hazards, and to tailor the laboratory's training program so that trainees learn to recognize and properly mitigate these risks. The goal should be for each trainee to receive a fundamental level of training in the process of risk assessment for the BSL-3 or ABSL-3 and to understand the tools at his or her disposal to control hazards, which include technical proficiency, good laboratory practices, and integrity of containment equipment and facility safeguards.
The BMBL provides a method for conducting a risk assessment. Biological risk assessment is a subjective process requiring consideration of many hazardous characteristics of agents and procedures, with judgments often based on incomplete information. There is no standard approach for conducting a biological risk assessment, but some structure can be helpful in guiding the process. The BMBL risk assessment procedure involves 5 steps.
1. Identify agent hazards and perform an initial assessment of risk. The principal hazardous characteristic of an agent includes its capability to infect and cause disease in a susceptible human host, the severity of disease, and the availability of preventive measures and effective treatments.
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 a hazard.
3. Make a determination of the appropriate biosafety level and select additional precautions indicated by the risk assessment. The 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. Other issues such as a laboratory worker's susceptibility to disease need to be addressed. Consultation with an occupational physician knowledgeable in infectious diseases is advisable in these circumstances.
4. Evaluate the proficiencies of staff regarding safe practices and the integrity of safety equipment. 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 that they have developed good habits that sustain excellence in the performance of those practices.
5. Review the risk assessment with a biosafety professional, subject matter expert, and institutional review groups, such as the IBC. A review of the risk assessment and selected safeguards by knowledgeable individuals is always beneficial and sometimes required by regulatory or funding agencies, as is the case with the NIH Guidelines. Review of potentially high-risk protocols by the local IBC should become standard practice. Adopting this step voluntarily will promote the use of safe practices in work with hazardous agents in microbiological and biomedical laboratories.
A risk assessment is a standard practice that laboratory directors, principal investigators, and biosafety professionals perform to identify the potential risks of handling infectious agents in the laboratory and to assess the proficiency of laboratory workers in the use of selected safe practices and containment equipment designed to control risks. The capability of the laboratory staff to control hazards will depend on the training, technical proficiency, and safe work habits of all members of the laboratory and the operational integrity of the containment equipment and facility safeguards.
Table 1 emphasizes the importance of conducting risk assessments to identify the hazardous characteristics of agents that are handled in BSL-3 and ABSL-3 facilities and for ensuring that the principal investigator and other laboratory scientists whose research will involve the handling of the listed select agents and toxins have acquired the competence required to contain and prevent exposures to the agents. The agents listed represent the bacteria and viruses that have been the causative agents for most laboratory-acquired infections recorded since David Bruce isolated Brucella melitensis in the late 19th century. Harding and Byers 12 listed the following records of laboratory-acquired infections occurring between 1930 and 2004: Brucella spp. (569), Coxiella burnetii (457), Francisella tularensis (228), and Venezuelan equine encephalitis virus infections (146). The low infectious dose of these agents significantly increases their risk to laboratory workers. In addition, CDC has reported laboratory-acquired infections from the handling of Brucella melitensis and Francisella tularensis between 2003 and 2009.
BSL-3 Containment Recommendations for Select Agents and Toxins 8
BSL-3 practices recommended for handling materials from which B virus is being cultured using appropriate personal protective equipment.
BSL-3 practices and equipment recommended for activities with a high potential for aerosol or droplet production.
BSL-3 containment recommended for propagating and manipulating sporulating cultures already identified as Coccidioides spp. and for processing soil or other environmental materials known to contain infectious arthroconidia.
BSL-3 containment (high risk: low inhalation infective dose <10 infectious particles).
BSL-3 containment (high risk: infective dose unknown; respiratory protection such as PAPR recommended; BSL-3Ag ABSL-3 may be required for research occurring in nonendemic areas).
BSL-2 facilities with BSL-3 practices for vaccinated personnel performing other work with monkeypox virus; ABSL-3 work in experimentally or naturally infected animals.
Reconstructed replication competent forms of the 1918<pandemic influenza virus containing any portion of the coding regions of all 8 gene segments (reconstructed 1918 influenza virus) BSL-3; ABSL-3; HEPA-filtered exhaust air and use of respiratory protection (particularly negative-pressure HEPA-filtered respirators or PAPRs) also required.
BSL-2 containment for clinical sample testing and diagnostic quantity manipulation; BSL-3 containment for larger quantities and/or aerosol generation.
BSL-3 facilities and arthropod containment level 3 practices are recommended for all laboratory work involving infected arthropods.
Conclusion
BSL-3 laboratories are unique work environments that must always be staffed with highly qualified workers in order to properly manage risks posed by the infectious agents being used and ensure compliance with all applicable safety standards and guidelines. This article outlined a methodical approach to the proper preparation of workers assigned to the high-containment environment. Only those who have successfully completed a comprehensive training program, including demonstration of proficiency while working with a mentor, should be allowed to perform unsupervised work activities.
The quality of the safety training program is the foundation of safety and compliance, and a commitment to excellence must always be the goal. Given that the employer is ultimately responsible for providing a safe work environment, it is critical that senior administrators recognize and support this initiative and dedicate the significant resources and time that are required to carry it to fruition. 13
References
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