Laboratory Safety Manual Section 2 Biological Safety
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Section 2 Biological Safety Chapter 1 Introduction Laboratory Safety Manual Section 2 Biological Safety INTRODUCTION PURPOSE This section has been prepared to provide the laboratory personnel at Duke University with the information necessary to protect them and the surrounding environment from hazards associated with the use of biological materials. The guidelines which follow provide a means for evaluating the risks of work involving biological materials and introduce the proper handling practices which will minimize the risk of an occupational acquired infection. History has shown that if not handled appropriately, infectious agents can be transmitted to laboratory employees, and rarely, to people outside of the laboratory. Biohazardous materials are those which are either known to cause, or that present a potential risk to the health of humans or animals. Such materials would include, but are not limited to: bacteria, fungi, viruses, parasites, rickettsia, rDNA toxins, human blood and unfixed human tissues or cell lines. Work with biohazardous material(s) requires consultation with the OESO – Biological Safety Division (919-684-8822). Examples for Safe Handling of Biohazardous Materials Material Precautions/ Employee Health Waste Disposal Shipping Training Containment Services Requirements Human- • BBP Exposure • Hepatitis B • All waste • Classify and • New derived Control Plan (see vaccine decontaminated package Employee (e. g. Appendices for this according to Duke according to I Orientation blood, section) • All exposures Medical Waste ATA’s DGR body reported to Management Policy. • General Lab fluids, • BSL 2 (see BMBL* EOHW Exposure • Receipt or Safety (annual tissue, or "Biosafety Hotline, 115 (on shipment of updates) cells, Levels" of this campus) and 684- “select agents” etc.) section) 8115 (off must comply • Shipping campus) with 42 CFR Part training every 2 • BSL2 SOP 73. years • Complete the Report of • Importation of • BSL2 training Work Related infectious Injury or Illness substances • Bloodborne form. requires a Pathogens permit. training P a g e | 1 Section 2 Biological Safety Chapter 1 Introduction Material Precautions/ Employee Waste Disposal Shipping Training Containment Health Services Requirements Known (or • Refer to BMBL* • Contact • All waste • Classify and New Employee suspect) or "Biosafety EOHW at 684- decontaminated package Orientation human or animal Levels" of this 3136 for according to Duke according to I pathogens (e. g. section for vaccination Medical Waste ATA’s DGR • General Lab HIV, appropriate BSL information. Management Safety (annual Rickettsial or ABSL. Policy. • Receipt or updates) agents, • All exposures shipment of Salmonella, etc.) • Refer to reported to “select agents” • Shipping infectious agent EOHW Exposure must comply training every 2 PSDS** Hotline, 115 (on with 42 CFR years campus) and Part 73. •BSL2 SOP 684- 8115 (off • BSL2 training campus) • Importation of infectious • Complete the substances Report of Work requires a Related Injury permit or Illness form. Recombinant • Follow NIH • Contact • All waste • Classify and • New DNA/Transgenics Guidelines (also EOHW at 684- decontaminated package Employee see 3136 for according to Duke according to I Orientation "Recombinant vaccination Medical Waste ATA’s DGR DNA" in this information. Management • General Lab section) Policy. Safety (annual • Visit Duke's IBC • All exposures updates) website to to human complete pathogens are • Shipping appropriate reported to training every 2 registration EOHW Exposure years • If material is a Hotline, 115 (on biological vector, campus) and use appropriate 684-8115 (off BSL for wild type campus). agent. See Exposures to rDNA/viral vector rDNA are webpage for reported to more information OESO-Biological and the rDNA Safety 684- registration form 8822. for IBC registration • Complete the information. Report of Work If BSL2 or above, Related Injury must have SOP. or Illness form. * Biosafety in Microbiological and Biomedical Laboratories, 5th ed., CDC/ NIH, 2009: (https://www.cdc.gov/labs/BMBL.html) ** Health Canada, Laboratory Center for Disease Control; Office of Biosafety: (http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php) P a g e | 2 Section 2 Biological Safety Chapter 2 Routes of Exposure ROUTES OF EXPOSURE There are four main routes of exposure that one must try to avoid when working with biohazardous agents in the laboratory. These would include percutaneous injuries, inhaling infectious aerosols, exposure to mucous membranes, and ingestion. Percutaneous injuries Percutaneous injuries can result from needlesticks, cuts or abrasions from contaminated items. These exposures are particularly serious because of the potential for immediate entry of the agent into a normally sterile bloodstream. All sharps items should be handled and disposed of as noted in the Waste Management section. Inhalation of aerosols Many laboratory procedures can cause the aerosolization of infectious agents. Some of these procedures include the use of vortexes, blenders and sonicators. Proper work practices must be implemented to minimize the aerosolization of all materials, especially those which are known to be transmitted by the aerosol route (e.g., Adenovirus, Vaccinia virus, Mycobacterium tuberculosis, etc.). See Chapter 7, Laboratory Equipment for more information about minimizing and containing aerosols in the laboratory. Mucous membrane Exposure of mucous membranes to infectious agents can lead to occupationally acquired infections. Mucocutaneous exposures can result from splashes to the eyes, nose or mouth, or by inadvertent inoculation via contaminated hands. Face protection should always be used if there is a likelihood of splash or splatter. Ingestion Accidental ingestion of biohazardous materials can result from improper personal hygiene in the laboratory. Food and drink are prohibited in all areas of the laboratory in which work is conducted with potentially infectious materials. Hands must always be washed before leaving the laboratory, and immediately if visible contamination occurs. P a g e | 3 Section 2 Biological Safety Chapter 3 Standard Laboratory Practice and Technique STANDARD LABORATORY PRACTICE AND TECHNIQUE Biosafety Levels and Warning Signage Four biosafety levels (BSLs) are summarized in the table below for proper handling of biohazardous materials. BSLs consist of combinations of laboratory practices and techniques, safety equipment, and laboratory facilities. Each combination is specifically appropriate for the operations performed the documented or suspected routes of transmission of the infectious agents, and for the laboratory function tor activity. BSL Agents Practices Safety Equipment Facilities (Primary Barriers) (Secondary Barriers) 1 Not known to consistently Standard None required Open bench top, sink required cause diseases in microbiological immunocompetent adult practices humans 2 Associated with human BSL-1 practices plus: Primary barriers: Class I or BSL-1plus: disease. Hazard: • limited access II biosafety cabinets or • non-fabric chairs and other percutaneous injury, • biohazard warning other physical furniture easily cleanable mucous membrane signs containment devices used • autoclave available exposure, ingestion • sharps precautions for all manipulations of • eyewash readily available • biosafety manual agents that cause splashes defining waste or aerosols of infectious decontamination or materials; PPE: laboratory medical surveillance coats, gloves, face policies protection as needed 3 Indigenous or exotic agents BSL-2 practices plus: Primary barriers: Class I or BSL-2 plus: with potential for aerosol • controlled access II biosafety cabinets or • physical separation from access transmission; disease may • decontamination of other physical corridors have serious or lethal all wastes containment devices used • hands-free handwashing- sink consequences • decontamination of for all manipulations of • self-closing double door access lab clothing before agents; PPE: laboratory • exhaust air not recirculated laundering coats, gloves, respiratory • negative airflow into laboratory • baseline serum protection as needed • eyewash readily available in lab 4 Dangerous/exotic agents BSL-3 practices plus: Primary barriers: All BSL-3 plus: which pose high risk of life- • clothing change procedures conducted in • separate building or isolated zone threatening disease, before entering Class III biosafety cabinets • dedicated supply/exhaust, vacuum aerosol-transmitted lab • shower on exit or Class I or II biosafety and decon system infections; or related • all material cabinets in combination agents with unknown risk decontaminated on with full-body, air supplied There are no BSL-4 labs at Duke of transmission exit from facility positive pressure suit Summarized from Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, 2009. http://www.cdc.gov/biosafety/publications/bmbl5/ P a g e | 4 Section 2 Biological Safety Chapter 3 Standard Laboratory Practice and Technique Biohazard Warning Signage A sign incorporating the universal biohazard symbol must be posted at the entrance to the laboratory when infectious agents are present. Biosafety Level 1 (BSL-1): The sign may include the name of the agent (s) in use, and the name and phone number of the laboratory supervisor or other responsible personnel. BSL-1 is suitable for work involving well-characterized agents not known to consistently cause disease in immunocompetent adult humans, and present minimal