Biological Safety in laboratories THE CHALLENGE Safe Labs • What is ‘safe’? • Who decides? • How to inform occupants? • How to manage change? LAB/RESEARCH COMMUNITY SAFETY Involves & relies on •Teaching •Laboratory •Research Laboratories Research Field •Radiation & Bio •Chemical •Fire Prevention, Building Design & Safety Officers Hygiene Officer 3 Maintenance, Industrial Hygienists etc. • EVERYONE is responsible for quality and safety SAFETY, HEALTH & ENVIRONMENTAL GOALS ZERO… • Workplace injuries • Workplace illnesses • Related injury/illness at home • Property loss • Unintended environmental damage MANAGEMENT OF HEALTH, SAFETY AND THE ENVIRONMENT . Achieving excellence in providing a healthy and safe working environment . Supporting environmentally sound practices in the conduct of University activities . Comply with all applicable health, safety, and environmental protection laws, regulations and requirements BIOLOGICAL RESEARCH IS ANY EXPERIMENTAL ACTIVITY INVOLVING THE FOLLOWING BIOLOGICAL AGENTS: • Human source materials (includes primary and immortal cell lines, clinical specimens, samples, tissues, etc) • Animals and animal source materials • Plants and plant source materials • Recombinant and synthetic DNA/RNA (including all viral vectors, RNAi technologies, etc). • Microorganisms (bacteria, viruses, fungi, prions, yeast, algae, rickettsiae, parasites, etc) • Biological toxins, allergens The CDC Biosafety in Microbiological and Biomedical Laboratories (BMBL) classifies risk group agents to: RG1 Agent not associated with disease in healthy adult humans e.g. Bacillus subtilis RG2 Associated with human disease which is rarely serious and preventive or therapeutic interventions are often available e.g.Staphylococcus aureus, influenza viruses type A, B, and C RG3 Serious or lethal human disease; preventive or therapeutic interventions may be available e.g. Mycobacterium tuberculosis •RG4 Serious or lethal human disease; preventive or therapeutic interventions are usually not available e.g. Ebola Quick question Which risk group do you think the avian flu virus would belong to? Resources for assigning risk group/biosafety level 1. The NIH Guidelines Appendix B assigns risk groups to some biological agents. 2. The American Biological Safety Association (ABSA) website provides a searchable database of biological agents and their assigned biosafety levels by country. BIOSAFETY LEVELS BL-1: Agents are not known to cause disease BL-2: Agents are associated with human disease BL-3: Agents are associated with human disease and are potentially transmitted as aerosols BL-4: Agents of life threatening nature BIOSAFETY LEVEL 1 (BL-1) Use BL-1 when working with: • Well characterized agents • Agents that are not known to cause disease in healthy humans • Agents that are of minimal hazard to lab personnel and the environment Examples of BL-1 Agents: E. coli JM109, DH5a Saccharomyces cerevisiae BIOSAFETY LEVEL 1 (BL-1) Standard Work Practices Use mechanical pipetting devices Wash hands frequently Minimize splashes and aerosols Decontaminate work surfaces daily Handle wastes properly Maintain insect and rodent control program BIOSAFETY LEVEL 1 (BL-1) Personal Protective Equipment (PPE) Lab coat or apron Safety glasses or goggles Gloves as needed BIOSAFETY LEVEL 2 (BL-2) Use BL-2 practices when working with: Agents of moderate potential hazard to personnel and the environment Examples of BL-2 agents: Human blood or body fluids E. coli 0157:H7 Clostridium botulinum Retroviral vectors Human cells in cell culture BIOSAFETY LEVEL 2 (BL-2) Requirements Adequate illumination Eyewash facility Negative air pressure Autoclave available Biological safety cabinet (BSC) Lab must be separated from public areas BIOSAFETY LEVEL 2 (BL-2) Standard Work Practices Use mechanical pipetting devices Wash hands frequently Minimize splashes and aerosols Decontaminate work surfaces daily Handle wastes properly Maintain insect and rodent control program BIOSAFETY LEVEL 2 (BL-2) Special Practices Place used slides and coverslips in sharps containers, never in any other receptacle. Sharps containers are: • Red in color • Marked with the biohazard symbol • Puncture resistant • Leak proof BIOSAFETY LEVEL 2 (BL-2) Personal Protective Equipment (PPE) • Lab coat or apron • Safety glasses or goggles • Gloves • Biosafety cabinet CASE STUDY CASE STUDY Opinion in favor of Dr Harran: 1. Why should he be responsible? Most professors don’t provide their graduate students much safety advice anyway. 2. Sangji should have worn a lab coat without the instruction of her advisor, it’s one of the basics of Organic chemistry and she is a graduate student not an undergraduate. 3. Sangji should have read the MSDS for tert butyl lithium, which states that she should not use a plastic syringeand should use a separate glass syringe for each transfer. CASE STUDY Opinion against Dr Harran: 1. Sangji did not have enough experience to perform the experiment on her own. 2. Dr Harran’s lab did not conform to safety codes. 3. Dr Harran should have advised Sangji to read the material safety data sheets for the chemicals she was using. LESSONS LEARNED Importance of safety audits and inspections by specialized personnel. Changing the culture, i.e. The system should help enforce safety by empowering students rather than dictating it from on high. BIOSAFETY CABINETS (BSC) Biosafety cabinets (BSC) control airborne contaminants during work with infectious material through the use of laminar airflow and high efficiency particulate air (HEPA) filtration. BIOSAFETY CABINETS (BSC) It is designed to protect personnel and the environment from infectious materials inside the cabinet and to protect the material inside the cabinet from contamination from the lab environment. BIOSAFETY CABINETS (BSC) Provide product, personal and environmental protection. Various classes of BSCs are available • Amount of air exhausted • Amount of employee protection •http://oregonstate.edu/ehs/LVSG-BSC- type BIOSAFETY CABINET (CLASS IIB2) BIOSAFETY CABINETS (BSCS) Sterile air is exhausted from BSCs through a HEPA filter. • HEPA filter can trap particles to 0.3 microns • Chemical vapors and fumes pass through HEPA filters BSC LOCATION IN THE LABORATORY Since the air curtain created at the front of the cabinet can be easily disrupted, a BSC should be located away from air supply registers, entrances, high traffic areas, and laboratory equipment, e.g. centrifuges, that create turbulence. SAFE AND EFFECTIVE USE OF THE BSC Before beginning work: a.Monitor alarms, pressure gauges, or flow indicators for any changes. b.Turn the cabinet on and let it run for 3-5 minutes. c.Wipe work surface with an appropriate disinfectant, e.g. 70% ethanol. d.Plan your work and place everything needed for the procedure inside the BSC. e.Wipe items with disinfectant before placing in BSC. SAFE AND EFFECTIVE USE OF THE BSC Avoid airflow disruption that could affect the level of protection provided by the BSC: a. Keep the BSC free of clutter, e.g. extra equipment and supplies b. Don’t place objects over the front air intake grille. c. Don’t block the rear air intake grille. d. Limit traffic in the area when the BSC is in use e. Make sure lab door is closed, and avoid opening and closing door if located near the BSC. f. Move arms slowly when removing or introducing items. g. Keep all materials at least 4 inches inside the sash. h. Don’t operate a Bunsen burner in the cabinet. SAFE AND EFFECTIVE USE OF THE BSC While working: a.Work as far to the back of the BSC workspace as possible. b.Segregate contaminated and clean items. Work from “clean to dirty.” c.Clean up all spills in the cabinet immediately. Allow cabinet to run for 3-5 minutes before resuming work. SAFE AND EFFECTIVE USE OF THE BSC After completing work: a. Wipe down all items with an appropriate disinfectant before removing. b. Remove all materials and wipe all interior surfaces with an appropriate disinfectant, e.g. 70% ethanol. c. Periodically decontaminate under work grilles. BIOSAFETY CABINETS (BSCS) Helpful Hints . Enter straight into the cabinet, avoid sweeping motions . Don’t place materials on the grill . Keep discard pan or bag inside the cabinet CASE STUDY Lessons learned: 1. Never add any water to an unmarked container. 2. Label everything with the common chemical name. 3. Always attach your waste label to the waste bottle before you add the waste and fill it out completely and accurately. PROTECTIVE PERSONAL EQUIPMENT (PPE) CONTAMINATION ROUTES •Ocular invasion •Inhalation •Ingestion •Skin penetration PERSONAL PROTECTIVE EQUIPMENT The Strategy of Last Resort Respiratory Protection Eye, Face, Hand, Foot Protective Clothing Hearing Head Protection 37 • The Occupational Safety and Health Administration (OSHA) requires that personal protective equipment (PPE) be selected, applies to work done with animals too Eye/face/mucous membrane protection When working with potentially infectious materials outside of a biological safety cabinet mucous membranes must be protected by wearing a surgical-type with safety glasses Foot/skin protection Open toed shoes, sandals and other open footwear is prohibited. Shorts are forbidden in laboratories using potentially infectious materials. Protective lab coats are to be worn while in the laboratory. This protective clothing is removed and left in the laboratory before leaving for non-laboratory areas (e.g., cafeterias, library and administrative offices.) PERSONAL PROTECTIVE EQUIPMENT Hand protection GLOVES ARE NEVER A SUBSTITUTE FOR THOROUGH HAND WASHING