International Journal of Antimicrobial Agents 36S (2010) S70 S74 Contents lists available at ScienceDirect International Journal of Antimicrobial Agents journal homepage: http://www.elsevier.com/locate/ijantimicag Biosafety and biosecurity measures: management of biosafety level 3 facilities Adel N. Zaki US Naval Medical Research Unit No. 3, PSC 452, Box 5000, FPO AE 09835-9998 article Keywords: Biosafety Biosecurity info abstract With the increasing biological threat from emerging infectious diseases and bioterrorism, it has become essential for governments around the globe to increase awareness and preparedness for identifying and containing those agents. This article introduces the basic concepts of laboratory management, laboratory biosafety and laboratory biosecurity. Assessment criteria for laboratories biorisk should include both biosafety and biosecurity measures. The assessment requires setting specific goals and selecting management approaches. In order to implement technologies at the laboratory working level, a management team should be created whose role is to implement biorisk policies, rules and regulations appropriate for that facility. Rules and regulations required by government authorities are presented, with special emphasis on methods for air control, and liquid and solid waste management. Management and biorisk measures and appropriate physical facilities must keep pace, ensuring efficient facilities that protect workers, the environment, the product (research, diagnostic and/or vaccine) and the biological pathogen. Published by Elsevier B.V. on behalf of International Society of Chemotherapy. 1. Introduction The principal characteristics of an agent that render it hazardous are its capability to infect and cause disease in a susceptible human or animal host, its virulence as measured by the severity of the disease it causes, and the availability of preventive measures and effective treatments for the disease. The World Health Organisation (WHO) has recommended an agent classification for laboratory use that describes four general risk groups based on these principal characteristics and the route of transmission of the natural disease. The four groups address the risk to both the laboratory worker and the community. The US National Institutes of Health (NIH) guidelines establish a comparable classification and assign human aetiological agents to one of four risk groups on the basis of hazard. The descriptions of the WHO and NIH risk group classifications are given in Table 1. They correlate with, but do not equate to, biosafety levels. Determining the risk group (RG) of a biological agent is part of the biosafety risk assessment and helps in assigning the correct biosafety level (BSL) for containment (see Section 2.4). In general, RG2 agents are handled at BSL2 and RG3 agents at BSL3. However, the use of certain RG2 agents in large quantities might require BSL3 The views expressed in this article are those of the author and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, or the US Government. Tel.: +1 201 234 60425; fax: +1 202 234 80250. E-mail address: adel.zaki.ctr.eg@med.navy.mil. conditions, whereas some RG3 agents may be safely manipulated at BSL2 under certain conditions [1,2]. The protection of laboratory workers, the environment, the product and the biological agents is achieved by addressing laboratory management, laboratory biosafety measures and laboratory biosecurity measures. 2. Laboratory management Highly infectious biological material includes samples such as human or animal tissues, body fluids and vectors that are known to carry certain viruses or bacteria. These are usually collected from field sites in endemic areas or during outbreaks of known or unknown aetiology. Samples can also be collected from hospitals for routine laboratory testing. It is important to use appropriate precautions when handling these samples. 2.1. Sample collection General guidelines include the following: (1) select the proper anatomical site from which to collect the specimen; (2) avoid contamination with indigenous flora; (3) use approved sampling procedures, including appropriate sample collection devices; (4) collect a sufficient volume of material to enable all requested tests to be performed; (5) label each specimen with the patient s name, identification number, source of specimen, date and time of collection, and the initials of the collecting individual and (6) use a specimen container designed to promote the survival of the pathogenic agent [3 6]. 0924-8579/$ see front matter. Published by Elsevier B.V. on behalf of International Society of Chemotherapy. doi:10.1016/j.ijantimicag.2010.06.026
A.N. Zaki / International Journal of Antimicrobial Agents 36S (2010) S70 S74 S71 Table 1 WHO and NIH risk group classifications [1,2]. Risk group Individual risk Community risk Description 1 Low Low A microorganism that is unlikely to cause human or animal disease 2 Moderate Low A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment Laboratory exposure may cause serious infection but effective treatment and preventive measures are available and the risk of spread of infection is limited 3 High Low/moderate A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another Effective treatment and preventive measures are available 4 High High A pathogen that usually causes serious human or animal disease and can be readily transmitted from one individual to another, directly or indirectly Effective treatment and preventive measures are not usually available 2.2. Sample shipping Shipping regulations for biological material differ from those for infectious substances. Infectious substances fall into two categories for shipping purposes. A Category A substance is one that is transported in a form that, when exposure to it occurs, is capable of causing permanent disability, life-threatening or fatal disease in otherwise healthy humans or animals. A Category B infectious substance is one that does not meet the criteria for Category A. Because of the difference in the hazard posed by Category A and Category B infectious substances there are variations in the packaging, labelling and documentation requirements for each category. The packaging requirements are determined by standardised regulations, such as biosecurity regulations, export and import controls, IATA dangerous goods regulations, UN Model Regulations on the Transport of Dangerous Goods, regional, national shipping, local shipping, mode of transportation and postal regulations, carrier requirements and WHO recommendations, all of which are subject to change and regular upgrade [1,3 6]. Shippers of infectious substances must ensure that packages are prepared in such a manner that they arrive at their destination in good condition and present no hazard to persons or animals during transport. Blood specimen bottles and tubes should be transported upright and secured in a screw-capped container or in a rack in a transport box. They should have enough absorbent paper around them to soak up all the liquid in case of spillage. If the specimen will reach the laboratory within 24 h, most pathogens can be recovered from blood cultures transported at ambient temperature. For longer transit periods samples should be kept at 4 8 C unless it is a cold-sensitive bacterial pathogen. 2.3. Sample labelling and identification The information contained in the case investigation and laboratory request forms is collected along with the specimen. Each patient should be assigned a unique identification number, usually known as the study number, by the study/laboratory coordinator. The identification number and the patient name should be present on all specimens, clinical forms and the laboratory request. All laboratory results and records will be reported using this identification number. 2.4. Laboratory handling Laboratory facilities are designated as basic biosafety level 1; basic biosafety level 2; containment biosafety level 3; and maximum containment biosafety level 4. Biosafety level designations are based on a composite of the design features, construction, containment facilities, equipment, practices and operational procedures required for working with agents from the various risk groups [1]. Agents are classified according to their pathogenicity, virulence, host range, communicability, transmission and environmental stability, and additional factors such as their toxicity or tendency to induce allergic reactions [1 5]. Fig. 1 provides a summary of the risk assessment strategy that should be followed to determine the correct biocontainment policy [7]. A laboratory management system is a set of interrelated elements used to establish policy and objectives and to achieve those objectives. Such a system includes organisational structure, planning activities (including, for example, risk assessment and the setting of objectives), responsibilities, practices, procedures, processes and resources. The requirements for a BSL3 research management system are given in Table 2. 3. Laboratory biosafety A laboratory biosafety plan is a set of preventive measures designed to reduce the risk of accidental exposure to or release of a biological hazard. A biosafety plan includes a set of interrelated elements covering work practice, the primary barrier (e.g. personal protective equipment and biosafety cabinets) and the secondary barrier, such as facility design features. Biosafety programmes should prevent employees and their families from acquiring laboratory-associated infectious diseases, prevent contamination of the environment, promote environmental quality and comply with all applicable national, international and local guidelines and regulations for the use of biohazards. A successful biosafety plan requires the establishment of a biosafety committee. The role of this committee is performing risk assessments, ensuring process, ensuring implementation and enforcement, establishing the biosafety level (facilities, equipment, practices and procedures), communicating hazards to employees, ensuring task-specific training, documenting and reviewing effectiveness, and encouraging strict adherence to safety policies. In addition, the committee advocates for resources, reviews research protocols and proposals, establishes occupational health programmes and inspects and reviews laboratory programmes. The biosafety plan also requires a biosafety officer, amongst whose duties are registering all work that carries a biological, chemical or radiological hazard; inspecting and evaluating all facilities; providing assistance to the scientific staff and providing training on institutional policy and basic safety. Safety is everyone s responsibility the institutional official(s), the biosafety officer, the laboratory managers, the laboratory staff and the support staff (maintenance and housekeeping). With this
S72 A.N. Zaki / International Journal of Antimicrobial Agents 36S (2010) S70 S74 Table 2 Requirements for the management of a biosafety level 3 (BSL3) facility. Fig. 1. Risk assessment strategy flow chart. Category Requirements Documents, policies and guidelines The organisation shall establish, document, implement and maintain a BSL3 management system in accordance with the requirements of the BSL3 laboratory guidelines (WHO, CDC, NIH, and/or BMBL) The organisation shall continually improve the effectiveness of the management programme through the use of the policy, objectives, self-audit programme, audit results, analysis of data, risk assessment, corrective and preventive actions and the management review Risk assessment and standard operating procedures (SOPs) The management programme shall have a completed risk assessment document, and all required SOPs such as (1) fire prevention and control, (2) hazard communication, (3) accident and incident reporting system, (4) respiratory protection programme, (5) chemical hygiene plan, (6) personal protective equipment use, (7) post-exposure and prophylaxis plan, (8) blood-borne pathogen control plan, (9) transfer of equipment, supplies and reagents in and out of BSL3, (10) disinfection and cleaning of work places, (11) maintenance and calibration of equipment, (12) entry and exit procedures, (13) donning and doffing of personal protective equipment Materials and waste management Materials: The organisation shall ensure that a review of the inventory is conducted at predetermined intervals based on risk, and at a level and frequency whereby materials can be accounted for in an appropriate manner Waste: The organisation shall ensure that the measures are put in place to minimise the quantities of biological waste. The organisation shall establish and maintain an appropriate waste management policy for biological agents and toxins A waste management policy shall: (1) ensure a programme is in place to minimise waste production, (2) ensure effective waste audit trails are in place and documented, (3) provide adequate facilities and procedures for the storage of waste (including short-term storage), (4) ensure methods are available for effective segregation and decontamination of mixed waste (e.g. infected animals that have received radioactive materials), (5) ensure appropriate packaging material is used to contain the waste and to maintain its integrity during storage and transportation Emergency response BSL3 management shall include emergency response plans for spills, injuries, fire, flood, equipment failure and power failure Records and plans Records of the following shall be kept: (1) risk assessment record, (2) BSL3 organisation chart, (3) position/job descriptions duties and responsibilities, (4) building drawings (layout, plumbing, mechanical and electrical), (5) cubic feet per minute measurements for supply and exhaust, (6) negative pressure records, (7) volume of each space within the BSL3, (8) air change per hour, (9) electrical load calculations, (10) electrical critical load, (11) emergency power load calculations, (12) list of equipment and electrical characteristics, and (13) single line diagram for the electrical distribution The following plans shall be developed: (1) occupational health (including consultation with employee physician, baseline serum banked, immunisations, knowledge of signs and symptoms); (2) safety plan; (3) decontamination plan (individual space, facility (BSL3 suite), high-efficiency particulates air filters, biological safety cabinet decontamination); (4) flow plans for people, materials, large equipment and emergencies; (5) security plan; (6) training plan Training and operation manuals Training for BSL3, laboratory safety, animal care, blood-borne pathogens, facility orientation and hands-on experience with a similar agent Operation: (1) operation of building management system, (2) operation of heating, ventilating and air conditioning system and control, (3) operation of emergency generator, (4) preventive maintenance operation programme, (5) annual maintenance operation, (6) operation of autoclave, (7) operation of biosafety cabinets, (8) operation of gloves box, and (9) operation of other equipment
A.N. Zaki / International Journal of Antimicrobial Agents 36S (2010) S70 S74 S73 Table 3 Types of waste to be controlled in a biosafety level 3 facility. System Air control system Infectious liquid waste from: Laboratory sinks Biological safety cabinet Shower room Toilet (if included in the facility) Infectious solid waste HEPA, high-efficiency particulate air. Control method (1) Individual supply and exhaust air ventilation system (2) System maintains differential and directional airflow (3) Assure flow inward from areas outside of the facility towards areas of highest potential risk within the facility (4) Supply and exhaust air flow is interlocked to ensure inward or zero airflow at all times (5) Use of HEPA filters (1) Steam heating decontamination system: based on decontamination using steam as a heat source. Liquid waste decontamination system and HEPA filtration should be considered for BSL3 facilities based on the risk assessment. The steam system should be a duplex process for safety considerations (2) Chemical disinfection system: most laboratory waste streams are effectively treated in a very short time, and thus excessive retention times are generally not required when using active systems. Systems should utilise sufficiently sophisticated controls to match chemical injection to the influent requirements and influent and effluent characteristics (1) Collection infectious waste must be collected immediately after it is generated, placed in appropriate containers and labelled and identified (2) Sterilisation (decontamination) the first phase of disposal a. Autoclaving is the procedure of choice for all decontamination processes b. Chemical disinfectants: some common disinfectants are active against most microorganisms (3) Disposal a. Incineration is the method of choice for final disposal of contaminated waste, even if laboratory practice requires that it is autoclaved first b. Recycling in the daily use not all equipment will require actual removal or destruction. The remainder will be recycled for use within the laboratories, including glassware, instruments and laboratory clothing. The approved procedure for recycling is decontamination by disinfecting and/or autoclaving in mind the biosafety (biorisk) programme should be written by senior management, scientific staff, safety personnel, security managers, emergency response personnel, law enforcement, engineering/maintenance and animal care staff. Each programme is a living document that must periodically be evaluated and updated [1 6]. In general, three major factors should be controlled within BSL3 facilities. These are shown in Table 3. Laboratory waste treatment systems should be appropriately sized to the demands of the facility. Careful consideration must be given to liquid waste decontamination, including the type of system, the need for vent filtration, the deep seal trap to prevent the escape of biological aerosols, and the selection of piping materials [8]. For solid waste it is necessary to be able to determine the efficacy of sterilisation. Several methods are available, including autoclave tape, chemical indicators and biological indicators. The reason for autoclave failure is usually either failure of the operator or failure of the equipment. Mistakes made by the operator may include: (1) improper use of containers that block the access of steam to the load; (2) sealing the bag closed before autoclaving; (3) not adding water to the load before autoclaving; (4) autoclaving a bag of waste that is too large for the autoclave and (5) over-filling an autoclave bag. Mechanical failure must be attended to by a trained technician. 4. Laboratory biosecurity Laboratory biosecurity encompasses a set of preventive measures designed to reduce the risk of intentional removal (theft) of biological material. Today, the world is facing a new challenge in safeguarding public health from potential domestic or international terrorism involving the use of dangerous biological agents or toxins. Existing standards and practices may require adaptation to ensure protection from such hostile actions. In addition, all nations mandate increased security within the microbiological and biomedical community in order to protect biological pathogens and toxins from theft, loss or misuse. A biosecurity plan addresses the threat to human and animal health, the environment and the economy posed by deliberate misuse or release of a biological agent. Such a plan includes a network of interrelated elements. 4.1. Physical security The physical security of the facility covers such issues as access control, the security camera system and the intrusion alarm system. 4.2. Personnel security Personnel security includes security clearance (screening, identity verification, educational/professional credential verification, military service verification, national criminal checks and financial checks) and security interviews. 4.3. Material control and accountability Material control and accountability addresses the insider threat and directly involves the personnel who work with pathogens and toxins that could be attractive to bioterrorists. Control of material comprises both procedural and physical measures for both material and information relevant to control and accountability. Accountability requires one-to-one correspondence between materials and individuals, together with a system of records, reporting and audit.
S74 A.N. Zaki / International Journal of Antimicrobial Agents 36S (2010) S70 S74 4.4. Transport security Transport security addresses the transportation of biological material outside a restricted area such as a research, public health or diagnostic laboratory, or a vaccine production facility. The transport could be across international borders, within a country or within the same facility. Transporting biological material requires (1) determination of the requirements for shipping (material transfer agreements, authorised recipients, regulations); (2) preparation of the samples for shipping (classification, packaging, marking and labelling, documentation and transfer of the package to the carrier); and (3) shipping management (authorised receipt verification, incident response, tracking, records and access controls). 4.5. Information security Information security refers to issues such as the server firewall and computer passwords [9 11]. 5. Conclusions It is important to heighten awareness of the need of BSL3 laboratory management, laboratory biosafety and laboratory biosecurity measures. One way to address this would be to introduce the concept through education at universities, within both life sciences and engineering departments. Assessment of facilities requires setting specific goals and selecting management approaches. Creating rules and regulations, including penalties, and implementing strategy is the role of government ministries. The government, via regulatory bodies, should be able to certify laboratories with the appropriate biocontainment level, and the regulatory bodies must obviously be provided with sufficient stable resources to carry out this function and annually evaluate the laboratories for licence renewal. This is crucial to expanding the application of biosafety and biosecurity technology throughout biological research laboratories, as well as to encouraging the development of new technologies. At the laboratory level, in order to implement appropriate technologies, a management team should be created whose role is to implement biorisk policies, rules and regulations required for that facility. Biorisk technology and physical facilities must keep pace to ensure efficient laboratories that protect workers, the environment, the product (research, diagnostic and/or vaccine) and the biological pathogen. Acknowledgments Dr Kenneth Earhart, Commanding Officer, US Naval Medical Research Unit #3 (NAMRU-3), Dr Mark Turner, Executive Officer, NAMRU-3, Dr Moustafa Mansour, Director, Research Science Directorate, NAMRU-3, Dr Emad Mohareb, Head Serology/Field Studies Unit, VZDRP, NAMRU-3, Mrs Salwa A. Mohanna, Administrative Support, NAMRU-3, and the US Department of State Biosecurity Engagement Program (BEP). Funding: AZ is an employee of the US Government. This article was prepared as part of the author s official duties. Competing interests: There is no conflict of interest. Ethical approval: Not required. References [1] WHO. Laboratory biosafety manual. 3rd ed. Geneva: World Health Organization; 2004 [accessed 25.03.2010] http://www.who.int/csr/resources/ publications/biosafety/who CDS CSR LYO 2004 11/en/index.html. [2] Centers for Disease Control and Prevention and National Institutes of Health. Biosafety in microbiological and biomedical laboratories. 4th ed. Washington: US Government Printing Office; 1999 [accessed 25.05.2010] http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm. [3] Public Health Agency of Canada. Laboratory biosafety guidelines. 3rd ed. Ottawa: Public Health Agency of Canada; 2004 [accessed 25.05.2010] http://www.phac-aspc.gc.ca/ols-bsl/lbg-ldmbl/index.html. [4] American Biological Safety Association. http://www.absa.org/ [accessed 25.05.2010]. [5] European Biosafety Association. http://www.ebsaweb.eu/ [accessed 25.05.2010]. [6] US Naval Medical Research Unit instructions. NAVMEDRSHUTHREEINST 5530.2: Implementation of command biological select agent and toxin possession, storage and use guidelines. NAVMEDRSHUTHREEINST 6260.1E: Biological safety. [7] European Committee for Standardization. CEN workshop 55 Guidance document for CWA 15793:2008 Laboratory Biorisk Management Standard. Brussels: European Committee for Standardization; 2008, http://www.cen. eu/cen/sectors/technicalcommitteesworkshops/workshops/pages/cwa15793- guide.aspx. [8] NIH Office of Research Facilities. Design policy and guidelines. Washington: National Institutes of Health; 2003 [accessed 25.05.2010] http://orf.od. nih.gov/policiesandguidelines/biomedicalandanimalresearchfacilitiesdesign PoliciesandGuidelines/HTMLVer/. [9] WHO. Biorisk management: laboratory biorisk guidance. Geneva: World Health Organization; 2006 [accessed 25.05.2010]. http://www.who.int/csr/ resources/publications/biosafety/who CDS EPR 2006 6.pdf. [10] Salerno RM, Gaudioso J. Laboratory biosecurity handbook. CRC Press; 2007. [11] Controlling biorisk manual. Sandia National Laboratories; 2009.