Biosafety Manual PDF - Lawrence Berkeley National Laboratory

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Biosafety Manual IBC-approved version (May 18, 2010) Disease Reservoir Vectors Causative Agent Exposure Routes Inhalation Ingestion Skin Contact tetanus animals Clostridium tetani — — wounds contaminated with dirt or objects containing animal or human feces or saliva various diseases such as skin infections or gastroenteritis fish aquarium water Mycobacterium marinum, M. fortuitum, Aeromonas hydrophila, other bacteria, and Cryptosporidium spp. protozoa — — skin contact with aquarium water, especially if skin has cuts or abrasions 3.4 Laboratory Procedure Hazards The BMBL five-step approach to assessing biological risk and selecting controls for laboratory work was initially presented in Section 3.2 of this manual. Step 2 (identifying laboratory procedure hazards) of this approach is discussed in this section. Historical data on laboratory acquired infections (LAIs) are an indicator of laboratory procedure hazards that have resulted in disease. Historical data show that past LAIs have occurred from: • parenteral inoculation by a contaminated sharp or syringe needle, • spills or splashes of contaminated materials directly onto the skin and mucous membranes, • ingestion through mouth pipetting, • animal bites and scratches, and • inhalation of infectious aerosol. See Section II of BMBL for more information regarding LAIs. Prevention of LAIs depends on the conscientious and proficient use of standard microbiological practices and special practices (see Section 4.1 of this manual) and the correct use of laboratory equipment. Table 7 below lists examples of hazards that may be found in laboratories using biological materials. Printed copies are not official versions of this manual. Before using the printed copy, verify that it is the most current version. 28
Biosafety Manual IBC-approved version (May 18, 2010) Table 7 Equipment Hazard Examples Equipment Type aerosol generating cryogenic temperatures high temperatures high pressure oxygen deficiencies rotational energies Hazards The diameter of aerosols generated from certain types of equipment will vary from 0.1 to 100 microns. • Bacterial cells and spores are 0.3 to 10 microns in diameter. • Viruses are 0.02 to 0.3 micron in diameter. • Biological particles generated from liquid or powder form particles that are 0.5 micron diameter. Cryogenic temperatures of –80°C are used to remove moisture from materials and contain low-temperature refrigerants. If protective equipment is not used, exposure to low temperature may cause cryogenic burns and frostbite. The use of heat to decontaminate or sterilize materials is widely used in the biological research laboratory. Physical injury from burns may occur from sudden accidental releases of heat sources or from the handling of hot items. Compressed gas cylinders and pressurized equipment are commonly used in the laboratory. Injury may occur from rupture high-pressure lines. Low-temperature freezers may include a backup system involving the use of a cryogenic liquid. Backup systems may consist of 50–200 liters of liquid nitrogen or liquid carbon dioxide under pressure. Liquid helium is also used in nuclear magnetic resonance (NMR) laboratories. Sudden release of such rotational energies can cause serious physical injury from unbalanced equipment or flying shrapnel. Examples • blender: 2 micron diameter particles • sonicator: 4.8 micron diameter particles • dropping bacterial flask: 3.5 micron diameter particles • dropping lyophilized culture: 10 micron diameter particles • pipette blow out: 4.9 micron diameter particles • vortex culture: 4.8 micron diameter particles • centrifuge: 4 micron diameter particles • freezers • lyophilizers (freeze dryers) • use of dry ice in shipping and receiving • dry heat temperatures used for sterilization range from 80°C to 200°C • wet heat is utilized by autoclaves to sterilize materials and can range between 80°C to 500°C • saturated steam operates at 121°C • autoclaves operate at high pressures of 1,000 kilo Pascal (145 psig) Oxygen deficiency environment may result from: • the displacement of oxygen by expanding gases (i.e., 700 parts of air to 1 part liquid nitrogen), • the linear displacement of oxygen from carbon dioxide (gas) generated from the use of dry ice, and • compressed gas cylinders or tanks. Tabletop and floor-mounted low, high, and ultracentrifuges rotate at speeds ranging from less than 5,000 to more than 100,000 rpm with rotor masses up to several kilograms. Printed copies are not official versions of this manual. Before using the printed copy, verify that it is the most current version. 29
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