Laboratory Biosafety Practices

1. Laboratory Biosafety Practices A. I. Bhat Indian Institute of Spices Research Calicut 673012, Kerala

2. Why Biosafety Practices? Protection: • workers • “products” • co-workers • lab support personnel • environment

3. What are the general hazards in a laboratory? • Fire • Breakage of glassware • Sharps • Spillages • Pressure equipment & gas cylinders • Extremes of heat & cold • Chemical hazards • Biological hazards • Radiation And many more!

4. worldwide literature search from 1979-2004 • 1,141 overt infections, 24 deaths • Most common causative agents of overt infection include: 1. M. tuberculosis 6. HBV 2. Arboviruses 7. Shigella spp. 3. C. burnetti 8. Salmonella spp 4. Hantavirus 9. HCV 5. Brucella spp. 10. N. meningitidis • Many asymptomatic infections • Many newer agents, e.g. SARS-CoV, Cryptosporidium, etc. • No specific exposure event in most cases __________________________________________________________WSLH

5. What were the predominant means of exposure? • As mentioned before no specific exposure event identified in most cases • Those identified included: • Inhalation of aerosols generated by work practices or procedures or spills • Percutaneous inoculation • Contamination of mucous membranes • Ingestion __________________________________________________________WSLH

6. Objective of biosafety is containment of potentially harmful biological agents • Purpose of containment is to reduce/eliminate exposure of lab workers, other persons and outside environment to biohazardous agents • Key elements of containment include: • Laboratory practice and technique • Safety equipment (primary barriers and PPE) • Facility design and construction (secondary barriers) • Risk Assessment of the work to be done with a specific agent or under specific circumstances determines the appropriate combination of these elements to employ __________________________________________________________WSLH

7. General Lab Requirements: Safety • Knowledgeable supervisor • Knowledgeable personnel • Aware of potential hazards • Proficient in practices & • techniques • Lab specific biosafety manual

11. Laboratory safety

12. Special procedures • Decontaminate work surfaces • Report spills and accidents • No animals in laboratories

18. Common Sense Rules • Wash hands & remove protective clothing before eating, drinking, smoking, handling contact lenses, applying lip balm or cosmetics • Keep hands away from eyes, nose, mouth while cleaning • Frequent hand washing is best defense against spreading infection

19. Elements of Containment Safety Equipment (Primary barriers and PPE) • Available for each possible route of exposure • Aerosol: BSCs, covered centrifuge carriers, loop incinerators or disposable loops, PPE • Percutaneous: sharps disposal; retractable needles • Mucous membrane contact: goggles or safety glasses, face shields, gloves • Ingestion: automatic pipetting devices __________________________________________________________WSLH

20. Elements of Containment The Biological Safety Cabinet The single most important piece of safety equipment in the laboratory! • Importance of the Biological Safety Cabinet • The principal device used to provide containment of infectious splashes or aerosols generated by many microbiological procedures • Provides protection to the operator, the laboratory environment and work materials • Which type is for you? __________________________________________________________WSLH

21. Biosafety Levels • 4 biosafety levels • Consist of combinations of lab practices and techniques, safety equipment and lab facilities • Purpose: To categorize risk associated with infectious agent and define the appropriate safety practices, equipment and facilities for handling the agent safely • Appropriate BSL determined by: • Microbiological agent Risk Group • Mode of transmission • Procedural protocols • Experience of staff • Likelihood of aerosol generation • Work involves use of amplified agent • Other? __________________________________________________________WSLH

22. Risk group classification

23. Risk Group 1 • Unlikely to cause animal or human disease • Non pathogenic agent

24. Risk Group 2 Pathogenic for humans Unlikely a serious hazard Treatment and preventive measures available Limited risk of spread of infection

25. Risk Group 3 Pathogenic, cause serious disease Effective treatment and preventive measures usually available Little person-to-person spread

26. Risk Group 4 Lethal, pathogenic agent Readily transmittable • direct, indirect Effective treatment and preventive measures not usually available

27. Risk groups, biosafety levels, practices and equipment

28. Recommended Biosafety Levels for Infectious Agents __________________________________________________________WSLH

29. Disinfection Disinfection requirements depend on the experimental work and nature of the agents being handled Contact time and concentration for disinfectants are specific for each material and manufacturer Efficacy is a function of surface, contact time, product, dilution

30. Using detergents • Use licensed detergents/disinfectants • Follow manufacturers’ recommendations • Wear adequate personal protective equipment • Perform hand hygiene • Avoid difficult to clean surfaces/ materials (carpet, wood) • Apply disinfectant to a large area - wiping, soaking • Avoid aerosolizing specimens while handling

31. Disinfection with household bleach Work areas • disinfect with 0.5% bleach after every procedure Contaminated supplies • soak with 0.5 % household bleach for 30 minutes • wash in soapy water • sterilize as necessary Photo: WHO

32. Risk Assessment • Whose responsibility? • Technically, the lab director • Practically, the bench microbiologist • Primary factors to consider: • Agent hazards • Laboratory procedures planned • Potential for aerosol generation • Consider facility, equipment needed; appropriate PPE • Capability of the staff • Training, technical proficiency, good habits • Known vs. unknown agent risk • For known or suspected agent, consult BMBL agent summary statements, other references • For unknown agent… __________________________________________________________WSLH

33. 1. Listen to or read instructions carefully before attempting to do anything. 2. Wear safety goggles to protect your eyes from chemicals, heated materials, or things that might be able to shatter. 3. Notify your teacher if any spills or accidents occur. General Safety Rules

34. 4. After handling chemicals, always wash your hands with soap and water. 5. During lab work, keep your hands away from your face. 6. Tie back long hair. General Safety Rules

35. 7. Roll up loose sleeves. 8. Know the location of the fire extinguisher, fire blanket, eyewash station, and first aid kit. 9. Keep your work area uncluttered. Take to the lab station only what is necessary. General Safety Rules

36. 10. It is suggested that you wear glasses rather than contact lenses. 11. Never put anything into your mouth during a lab experiment. 12. Clean up your lab area at the conclusion of the laboratory period. 13. Never “horse around” or play practical jokes in the laboratory. General Safety Rules

37. 1. Chipped or cracked glassware should not be used. Show it to the teacher. 2. Broken glassware should not be disposed of in a classroom trashcan. There is a special glass disposal container for it. 3. When pouring liquids into glassware, make sure the container you are pouring into is resting on a table at least a hands breadth from the edge. Glassware Safety

38. 4. If a piece of glassware gets broken, do not try to clean it up by yourself. Notify the teacher. 5. Do not place hot glassware in water. Rapid cooling may make it shatter. Glassware Safety

39. 1. Wear protective goggles whenever heating or pouring hazardous chemicals. 2. Never mix chemicals together unless you are told to do so (and then only in the manner specified). 3. Never taste any chemicals (you should never taste anything in the lab). Chemical Safety

40. 4. If you need to smell the odor of a chemical, waft the fumes toward your nose with one hand. Do not put your nose over the container and inhale the fumes. Chemical Safety

41. 5. Follow the instructions of your teacher when disposing of all chemicals. 6. Wash your hands after handling hazardous chemicals. Chemical Safety

42. 1. Use tongs and/or protective gloves to handle hot objects. 2. Never reach across an open flame or burner. Heating Safety

43. 3. Always point the top ends of test tubes that are being heated away from people. 4. When heating a test tube, move it around slowly over the flame to distribute the heat evenly. Heating Safety

44. 5. Only glassware that is thoroughly dry should be heated. 6. Heat glassware by placing it on a wire gauze platform on a ringstand. Do not hold it in your hand. Heating Safety

45. Injury: Burns To Do: Immediately flush with cold water until burning sensation is lessened. First Aid

46. Injury: Cuts, bruises To Do: Do not touch an open wound without safety gloves. Pressing directly on minor cuts will stop bleeding in a few minutes. Apply cold compress to bruises to reduce swelling. First Aid

47. Injury: The eyes To Do: Flush eyes immediately with plenty of water for several minutes. If a foreign object is lodged in the eye, do not allow the eye to be rubbed. First Aid

48. Spillages • Clear up spillage promptly • You will already have determined how to do this as part of your risk assessment • Dispose of any hazardous material as toxic waste Messy workers are usually poor workers!!

49. Skirts Shorts Open-toed shoes Sandals Jackets Bulky shirts or sweaters Floppy or loose sleeves Unacceptable Lab Clothing

50. Laboratory Equipment • Never use any laboratory equipment unless you are trained & have been authorised to do so • As well as injuring yourself you may cause very costly damage