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Title: Section 61-1.13 - Physical containment levels; P3 level

61-1.13 Physical containment levels; P3 level.

(a) Laboratory practices.

(1) Laboratory doors shall be kept closed while experiments are in progress.

(2) Work surfaces shall be decontaminated following the completion of the experimental activity, and immediately following spills of organisms containing recombinant DNA molecules.

(3) All laboratory wastes shall be steam-sterilized (autoclaved) before disposal. Other contaminated materials, such as glassware, animal cages, laboratory equipment, and radioactive wastes, shall be decontaminated by a method demonstrated to be effective before washing, reuse, or disposal.

(4) Mechanical pipetting devices shall be used; pipetting by mouth is prohibited.

(5) Eating, drinking, smoking, and storage of food are not permitted in the laboratory area in which recombinant DNA materials are handled.

(6) Persons shall wash their hands after handling organisms containing recombinant DNA molecules and when they leave the laboratory.

(7) Care shall be exercised to minimize the creation of aerosols. For example, manipulation such as inserting a hot inoculating loop or needle into a culture, flaming an inoculation loop or needle so that it splatters, and forceful ejection of fluids from pipettes or syringes shall be avoided.

(8) Contaminated materials that are to be steam-sterilized (autoclaved) or decontaminated at a site away from the laboratory shall be placed in a durable leak-proof container, which is closed before removal from the laboratory.

(9) Entry into the laboratory shall be through a controlled access area. Only persons who have been advised of the nature of the research being conducted shall enter the controlled access area. Only persons required on the basis of program or support needs shall be authorized to enter the laboratory. Such persons shall be advised of the nature of the research being conducted before entry, and shall comply with all required entry and exit procedures.

(10) Persons under 16 years of age shall not enter the laboratory.

(11) The universal biohazard sign shall be posted on the controlled access area door and on all laboratory doors when experiments requiring P3-level containment are in progress. Freezers and refrigerators or other units used to store organisms containing recombinant DNA molecules shall also be posted with the universal biohazard sign.

(12) An insect and rodent control program shall be instituted.

(13) Laboratory clothing that protects street clothing (e.g., long-sleeve solid-front or wrap-around gowns, no-button or slip-over jackets) shall be worn in the laboratory. Front-button laboratory coats are unsuitable. Laboratory clothing shall not be worn outside the laboratory and shall be decontaminated before it is sent to the laundry.

(14) Raincoats, overcoats, topcoats, coats, hats, caps, and such street outerwear shall not be kept in the laboratory.

(15) Gloves shall be worn when handling materials requiring P3 containment. They shall be removed aseptically immediately after the handling procedure and decontaminated.

(16) Animals and plants not related to the experiment shall not be permitted in the laboratory.

(17) Vacuum outlets shall be protected by filter and liquid disinfectant traps.

(18) Use of hypodermic needle and syringe shall be avoided when alternative methods are available.

(19) The laboratory shall be kept neat and clean.

(20) If experiments involving other organisms which require lower levels of containment are to be conducted in the same laboratory concurrently with experiments requiring P3-level physical containment, they shall be conducted in accordance with all P3-level laboratory practices.

(b) Containment Equipment.

(1) Biological safety cabinets shall be used for all equipment and manipulations that produce aerosols (e.g., pipetting, dilutions, transfer operations, plating, flaming, grinding, blending, drying, sonicating, shaking, centrifuging), where these procedures involve organisms containing recombinant DNA molecules, except where equipment design provides for containment of the potential aerosol.

(2) Laboratory animals held in a P3 area shall be housed in partial-containment caging systems, such as horsfall units, open cages placed in ventilated enclosures, solid-wall and -bottom cages covered by filter bonnets, or solid-wall and -bottom cages placed on holding racks equipped with ultraviolet radiation lamps and reflectors. (Note: Conventional caging systems may be used, provided that all personnel wear appropriate personal protective devices. These shall include, at a minimum, wrap-around gowns, head covers, gloves, shoe covers, and respirators. All personnel shall shower on exit from areas where these devices are required.)

(3) Alternative selection of containment equipment. Experimental procedures involving a host-vector system that provides a one-step higher level of biological containment than that specified can be conducted in the P3 laboratory using containment equipment specified for the P2 level of physical containment. Experimental procedures involving a host-vector system that provides a one-step lower level of biological containment than that specified can be conducted in the P3 laboratory using containment equipment specified for the P4 level of physical containment. Alternative combinations of containment safeguards are shown in Table I.

Table I


Classification of experiment Alternate combinations of physical and biological containment


Physical containment


Biological containment *


Laboratory design specified for:

Physical Containment

Laboratory practices specified for:


Containment equipment
specified for:

Biological containment

P3 HV2 P3 P3 P3 HV2
  HV2 P3 P3 P4 HV1
P3 HV1 P3 P3 P3 HV1
P3 HV1 P3 P3 P2 HV2

* See section 61-1.16 for description of biological containment.

(c) Special laboratory design.

(1) The laboratory shall be separated by a controlled access area from areas that are open to unrestricted traffic flow. A controlled access area is an anteroom, a change room, an air lock or any other double-door arrangement that separates the laboratory from areas open to unrestricted traffic flow.

(2) The surfaces of walls, floors, and ceilings shall be readily cleanable. Penetrations through these surfaces shall be sealed or capable of being sealed to facilitate space decontamination.

(3) A foot-, elbow-, or automatically operated hand-washing facility shall be provided near each primary laboratory exit area.

(4) Windows in the laboratory shall be sealed.

(5) An autoclave for sterilization of wastes and contaminated materials shall be available in the same building (and preferably within the controlled laboratory area) in which organisms containing recombinant DNA molecules are used.

(6) The laboratory shall have a ventilation system that is capable of controlling air movement. The movement of air shall be from areas of lower contamination potential to areas of higher contamination potential (i.e., from the controlled access area to the laboratory area). If the ventilation system provides positive pressure supply air, the system shall operate in a manner that prevents the reversal of the direction of air movement or shall be equipped with an alarm that would be actuated in the event that reversal in the direction of air movement were to occur. The exhaust air from the laboratory area shall not be recirculated to other areas of the building unless the exhaust air is filtered by HEPA filters or equivalent. The exhaust air from the laboratory area can be discharged to the outdoors without filtration or other means for effectively reducing an accidental aerosol burden provided that it can be dispersed clear of occupied buildings and air intakes.

(7) The treated exhaust-air from class I and class II biological safety cabinets may be discharged either to the laboratory or to the outdoors. The treated exhaust-air from a class III cabinet shall be discharged directly to the outdoors. If the treated exhaust-air from these cabinets is to be discharged to the outdoors through a building exhaust air system, it shall be connected to this system so as to avoid any interference with the air balance of the cabinet and the building ventilation system.


VOLUME A-1a (Title 10)