Appendix G specifies physical containment for standard laboratory experiments and defines Biosafety Level 1 through Biosafety Level 4. For large-scale (over 10 liters) research or production, Appendix K (Physical Containment for Large Scale Uses of Organisms Containing Recombinant DNA Molecules) supersedes Appendix G. Appendix K defines Good Large Scale Practice through Biosafety Level 3 - Large Scale. For certain work with plants, Appendix P (Physical and Biological Containment for Recombinant DNA Research Involving Plants) supersedes Appendix G. Appendix P defines Biosafety Levels 1 through 4 - Plants. For certain work with animals, Appendix Q (Physical and Biological Containment for Recombinant DNA Research Involving Animals) supersedes Appendix G. Appendix Q defines Biosafety Levels 1 through 4 - Animals.

Appendix G-I. Standard Practices and Training

The first principle of containment is strict adherence to good microbiological practices (see Appendices G-III-A through G-III-J, Footnotes and References of Appendix G). Consequently, all personnel directly or indirectly involved in experiments using recombinant DNA shall receive adequate instruction (see Sections IV-B-1-h, Responsibilities of the Institution--General Information, and IV-B-7-d, Responsibilities of the Principal Investigator Prior to Initiating Research). At a minimum, these instructions include training in aseptic techniques and in the biology of the organisms used in the experiments so that the potential biohazards can be understood and appreciated.

Any research group working with agents that are known or potential biohazards shall have an emergency plan that describes the procedures to be followed if an accident contaminates personnel or the environment. The Principal Investigator shall ensure that everyone in the laboratory is familiar with both the potential hazards of the work and the emergency plan (see Sections IV-B-7-d, Responsibilities of the Principal Investigator Prior to Initiating Research and IV-B-7-e, Responsibilities of the Principal Investigator During the Conduct of the Research). If a research group is working with a known pathogen for which there is an effective vaccine, the vaccine should be made available to all workers. Serological monitoring, when clearly appropriate, will be provided (see Section IV-B-1-f, Responsibilities of the Institution--General Information).

The Laboratory Safety Monograph (see Appendix G-III-O, Footnotes and References of Appendix G) and Biosafety in Microbiological and Biomedical Laboratories (see Appendix G-III-B, Footnotes and References of Appendix G) describe practices, equipment, and facilities in detail.

Appendix G-II. Physical Containment Levels

The objective of physical containment is to confine organisms containing recombinant DNA molecules and to reduce the potential for exposure of the laboratory worker, persons outside of the laboratory, and the environment to organisms containing recombinant DNA molecules. Physical containment is achieved through the use of laboratory practices, containment equipment, and special laboratory design. Emphasis is placed on primary means of physical containment which are provided by laboratory practices and containment equipment. Special laboratory design provides a secondary means of protection against the accidental release of organisms outside the laboratory or to the environment. Special laboratory design is used primarily in facilities in which experiments of moderate to high potential hazard are performed.

Combinations of laboratory practices, containment equipment, and special laboratory design can be made to achieve different levels of physical containment. Four levels of physical containment, which are designated as BL1, BL2, BL3, and BL4 are described. It should be emphasized that the descriptions and assignments of physical containment detailed below are based on existing approaches to containment of pathogenic organisms (see Appendix G-III-B, Footnotes and References of Appendix G). The National Cancer Institute describes three levels for research on oncogenic viruses which roughly correspond to our BL2, BL3, and BL4 levels (see Appendix G-III-C, Footnotes and References of Appendix G).

It is recognized that several different combinations of laboratory practices, containment equipment, and special laboratory design may be appropriate for containment of specific research activities. The NIH Guidelines, therefore, allow alternative selections of primary containment equipment within facilities that have been designed to provide BL3 and BL4 levels of physical containment. The selection of alternative methods of primary containment is dependent, however, on the level of biological containment provided by the host-vector system used in the experiment. Consideration will be given to other combinations which achieve an equivalent level of containment (see Sections IV-C-1-b-(1), Major Actions and IV-C-1-b-(2), Minor Actions).

Appendix G-II-A. Biosafety Level 1 (BL1) (See Appendix G-III-M, Footnotes and References of Appendix G)

Appendix G-II-A-1. Standard Microbiological Practices (BL1)

Appendix G-II-A-1-a. Access to the laboratory is limited or restricted at the discretion of the Principal Investigator when experiments are in progress.

Appendix G-II-A-1-b. Work surfaces are decontaminated once a day and after any spill of viable material.

Appendix G-II-A-1-c. All contaminated liquid or solid wastes are decontaminated before disposal.

Appendix G-II-A-1-d. Mechanical pipetting devices are used; mouth pipetting is prohibited.

Appendix G-II-A-1-e. Eating, drinking, smoking, and applying cosmetics are not permitted in the work area. Food may be stored in cabinets or refrigerators designated and used for this purpose only.

Appendix G-II-A-1-f. Persons wash their hands: (i) after they handle materials involving organisms containing recombinant DNA molecules and animals, and (ii) before exiting the laboratory.

Appendix G-II-A-1-g. All procedures are performed carefully to minimize the creation of aerosols.

Appendix G-II-A-1-h. In the interest of good personal hygiene, facilities (e.g., hand washing sink, shower, changing room) and protective clothing (e.g., uniforms, laboratory coats) shall be provided that are appropriate for the risk of exposure to viable organisms containing recombinant DNA molecules.

Appendix G-II-A-4-c. Laboratory furniture is sturdy. Spaces between benches, cabinets, and equipment are accessible for cleaning.

Appendix G-II-A-4-d. Each laboratory contains a sink for hand washing.

Appendix G-II-A-4-e. If the laboratory has windows that open, they are fitted with fly screens.

TABLE OF CONTENTS

APPENDIX G. PHYSICAL CONTAINMENT

Appendix G-I. Standard Practices and Training

Appendix G-II. Physical Containment Levels

Appendix G-II-A. Biosafety Level 1 (BL1) (See Appendix G-III-M, Footnotes and References of Appendix G)

Appendix G-II-A-1. Standard Microbiological Practices (BL1)

Appendix G-II-A-2. Special Practices (BL1)

Appendix G-II-A-3. Containment Equipment (BL1)

Appendix G-II-A-4. Laboratory Facilities (BL1)

Appendix G-II-B. Biosafety Level 2 (BL2)