Laboratory Biosafety Guidelines
2nd Edition 1996

CHAPTER 2

CONTAINMENT OF BIOHAZARDS

2.1 INTRODUCTION

Bacteria, viruses, fungi or other infectious agents are studied because they may cause disease, they can help us understand the natural world, and for many other reasons including the possibility of industrial applications. Since many of the agents can be pathogenic to humans, animals or other forms of life, their use poses risks which vary with each agent and the way it is used.

Safety standards are designed to reduce to an acceptable level the risks inherent in the use of dangerous materials. Stringent standards are set for hazardous agents and less stringent ones for agents which cause only minor problems. Safety standards are therefore compromises designed to allow needed work to proceed without exposing those involved or others to more than minimal risk.

2.2 SAFETY PRACTICES

The attitudes and actions of those who work in the laboratory determine their own safety, and that of their colleagues and of the community. Laboratory equipment and design can contribute to safety only if they are used properly by people who are genuinely concerned and knowledgeable about safety issues.

The Canada Labour Code requires that each employer provide safe working conditions and that employees be informed about all hazards they will face in the course of their duties. The employee is also given the right to withdraw from the workplace if faced with an unsafe condition. Other federal legislation such as Workplace Hazardous Materials Information System (WHMIS) requires that all hazardous substances, including microorganisms, be labelled in a specified manner and that there be a Material Safety Data Sheet (MSDS) available to accompany each hazardous substance. Employers are also required to provide all training necessary to work with the hazardous substance and to keep a written record of their employee education program. For more information, contact Labour Canada, tel: (613) 997-3520.

The following requirements are basic for any laboratory using infectious or toxic agents.

1. All laboratory personnel and others whose work requires them to enter the laboratory must understand the biological and other hazards with which they will come in contact through their normal work in the laboratory, and be trained in appropriate safety precautions and procedures. A laboratory safety manual must beprepared and adopted. And it is the responsibility of the laboratory director/principal investigator to ensure it identifies known and potential biohazards and specifies practices and procedures to eliminate or minimize such risks. The manual must also contain an emergency response plan. Personnel must be required to know, understand, and follow standard practices and procedures. Training in laboratory safety must be provided and competence in safe technique demonstrated before work is allowed with hazardous agents or toxins.
2. Laboratories should have a biological safety officer (BSO) and/or a biological safety committee whose responsibility include ensuring that all work is carried out in accordance with safety practices established at the Institution.

   The duties of the BSO should include providing technical advice on safety procedures and equipment, developing emergency plans, conducting safety inspections, providing biosafety training, conducting or supervising testing of containment systems, and providing guidance and information related to compliance with pertinent regulations.

3. The laboratory must be kept neat, orderly and clean, and storage of materials not pertinent to the work should be minimized.
4. Protective laboratory clothing (uniforms, coats, gowns) must be available and worn properly fastened by all personnel, including visitors, trainees and others entering or working in the laboratory. Protective laboratory clothing must not be worn in non-laboratory areas. Suitable footwear with closed toes and heels and preferably with non-slip soles must be worn in all laboratory areas.
5. Gloves must be worn for all procedures that might involve direct skin contact with toxins, blood, infectious materials, or infected animals. Rings or hand jewelry which would interfere with glove functioning should be removed before gloving. Gloves should be removed carefully and decontaminated with other laboratory wastes before disposal. Reusable gloves (e.g. insulated, chemical resistant, etc.) may be used only where necessary and must be appropriately decontaminated.
6. Safety face and eyewear (e.g. glasses, goggles, face shields, or other protective devices) must be worn when necessary to protect the face and eyes from splashes, impacting objects, harmful substances, UV light, or other rays.
7. Eating, drinking, smoking, storing food, personal belongings or utensils, applying cosmetics, and inserting or removing contact lenses are not permitted in any laboratory work area. Contact lenses should be worn only when other forms of corrective eyewear are not suitable. The wearing of jewelry should be discouraged in the laboratory.
8. Oral pipetting of any substance is prohibited in any laboratory.
9. Long hair must be tied back or restrained.
10. Hands must be washed after gloves are removed, before leaving the laboratory, and at any time after handling materials known or suspected to be contaminated.
11. Work surfaces must be cleaned and decontaminated with a suitable disinfectant at the end of the day and after any spill of potentially dangerous material. Loose or cracked work surfaces must be replaced or repaired.
12. All technical procedures must be performed in a manner that minimizes the creation of aerosols.
13. All contaminated or infectious liquid or solid materials must be decontaminated before disposal or reuse. Contaminated materials that are to be autoclaved or incinerated at a site away from the laboratory must first have the outside of the container disinfected chemically or be double-bagged.
14. Access to the laboratories must be strictly limited (containment Levels 3 and 4). Decisions on entry into containment Level 1 and 2 laboratories should be at the discretion of the laboratory director/principal investigator (e.g. only persons who have been advised of the potential hazards and meet any specific entry requirements such as immunization should be allowed to enter the laboratory area). Children under the age of 16 years old should not be permitted in the laboratory or support areas. Pregnant women or immunocompromised people who work in or enter the laboratory should be advised of the associated risks.
15. Hazard warning signs, indicating the risk level of the agents being used, must be posted outside each laboratory. Where infectious agent(s) used in the laboratory require special provisions for entry, the relevant information must be included in the sign. The agent must be identified, and the name of the laboratory supervisor and other responsible person(s) as well as any special conditions for staff entry must be listed.
16. The use of needles, syringes and other sharp objects should be strictly limited. Needles and syringes should be used only for parenteral injection and aspiration of fluids from laboratory animals and diaphragm bottles. Extreme caution should be used when handling needles and syringes to avoid autoinoculation and the generation of aerosols during use and disposal. Procedures should be performed in a biological safety cabinet. Needles should not be bent or sheared. They should not be replaced in the sheath or guard. They should be promptly placed in a puncture-proof container and decontaminated, preferably by incineration or autoclaving, before disposal.
17. All spills, accidents, and overt or potential exposures must be reported in writing to the laboratory supervisor or acting alternate as soon as circumstances permit; this person should file this report with management and the appropriate biosafety officer or committee. Appropriate medical evaluation, surveillance, and treatment should be sought and provided as required. Actions taken to prevent future occurrences should be documented..
18. Baseline serum for laboratory and other at-risk personnel (eg. laboratory support and maintenance staff) should be collected and stored. Additional serum specimens may be collected periodically, depending on the agents handled or the function of the facility.
19. Laboratory workers should be protected by appropriate immunization where possible. Levels of antibody considered to be protective should be documented. Particular attention must be given to individuals who are or may become immunocompromised, as vaccine administration may be different than for immunologically competent adults.

2.3 USE OF LABORATORY ANIMALS

Naturally occurring or experimentally induced infections in laboratory animals may be transmitted to other laboratory animals, invertebrates and laboratory workers. Animals infected or challenged experimentally with organisms in any of the risk groups may be small (e.g. mice) or large (e.g. livestock), have unique housing requirements (e.g. fish) or have uncharacterized susceptibilities. The requirements for maintenance of the animals may differ therefore in scale and degree but the basic principles for microbiological safety will be similar to those outlined in Section 2.2 and must be followed.

In addition, the following requirements and conditions must be satisfied:

• All aspects involved in the proposed use of animals in research must meet the standards and regulations for the care and maintenance of experimental animals as described by the Canadian Council on Animal Care, relevant provincial legislation and local animal care authorities.

• The appropriate species must be selected for animal experiments to reduce potential biohazards.

• The investigator and/or person(s) responsible for the animal experiment must ensure that all those having contact with the animals and waste materials are familiar with and aware of any special precautions and procedures that may be required. Where possible, personnel should be protected by immunization with appropriate vaccines.

• It is essential that all accidents, including animal bites and scratches or cuts from cages or other equipment, be reported and recorded.

• Small laboratory rodents or other small animals that escape from their cages should be killed when captured, their carcasses incinerated and the area should be fully decontaminated. In the event that animals escape the containment perimeter, the relevant authorities must be notified promptly and appropriate action initiated.

• Unexpected illness or deaths among animals must be reported to both the researcher and head of animal services without delay; instructions for dealing with such animals should be available. However, animals should not be touched until instructions are given by the person in charge.