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4. STANDARD OPERATING PROCEDURES

RYERSON UNIVERSITY. 2. Biological Safety Training Certificate Holder and User Training BIOLOGICAL SAFETY. . Infection ControlContainment of AerosolsWorking in CL 1 areasWorking in CL 2 areasBiological Safety CabinetsHuman Blood and Body FluidsNeedle Stick Injury Prevention. 4. Standard O

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4. STANDARD OPERATING PROCEDURES

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    1. RYERSON UNIVERSITY 1 4. STANDARD OPERATING PROCEDURES

    2. RYERSON UNIVERSITY 2 Biological Safety Training – Certificate Holder and User Training BIOLOGICAL SAFETY Infection Control Containment of Aerosols Working in CL 1 areas Working in CL 2 areas Biological Safety Cabinets Human Blood and Body Fluids Needle Stick Injury Prevention Signs and Labels Personal Protective Equipment Purchasing/Transfer Inventory Control Decontamination/Waste Disposal Large Scale Work

    3. RYERSON UNIVERSITY 3 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control

    4. RYERSON UNIVERSITY 4 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control

    5. RYERSON UNIVERSITY 5 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control – Hand Washing

    6. RYERSON UNIVERSITY 6 Frequent hand washing has proven to be the single most effective means of avoiding infection if done properly and frequently liquid dispensers should be used rather than bars Arguments that using antiseptic solutions cause pathogen resistance?Arguments that using antiseptic solutions cause pathogen resistance?

    7. RYERSON UNIVERSITY 7 Wet hands with warm water Dispense soap into a cupped hand Spread around hands and between fingers Wash hands for at least 10 sec. Rinse thoroughly under warm water. Dry hands thoroughly with paper towels.

    8. RYERSON UNIVERSITY 8 Continue on to: Containment of Aerosols

    9. RYERSON UNIVERSITY 9 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols

    10. RYERSON UNIVERSITY 10 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols

    11. RYERSON UNIVERSITY 11 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols

    12. RYERSON UNIVERSITY 12 Sterilization of inoculation loops in an open flame may create aerosols which may contain viable microorganisms. Use a shielded electric incinerator. Shorter handles minimize vibrations. Disposable plastic loops are good alternatives.

    13. RYERSON UNIVERSITY 13 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols

    14. RYERSON UNIVERSITY 14 Continue on to: Working in Containment Level 1 & 2 Areas

    15. RYERSON UNIVERSITY 15 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    16. RYERSON UNIVERSITY 16 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    17. RYERSON UNIVERSITY 17 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    18. RYERSON UNIVERSITY 18 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    19. RYERSON UNIVERSITY 19 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    20. RYERSON UNIVERSITY 20 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    21. RYERSON UNIVERSITY 21 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    22. RYERSON UNIVERSITY 22 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas

    23. RYERSON UNIVERSITY 23 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 2 areas

    24. RYERSON UNIVERSITY 24 Continue on to: Biological Safety Cabinets

    25. RYERSON UNIVERSITY 25 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    26. RYERSON UNIVERSITY 26 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    27. RYERSON UNIVERSITY 27 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Laminar Flow Cabinets

    28. RYERSON UNIVERSITY 28 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    29. RYERSON UNIVERSITY 29 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    30. RYERSON UNIVERSITY 30 Before using the cabinet: Ensure BSC is certified Disinfect work surfaces with disinfectant Place essential items inside cabinet Allow the blower to run for 5-10 min before work Certification sticker, with date of last certification, should be located on the front of the BSC. BSCs must be certified every year.Certification sticker, with date of last certification, should be located on the front of the BSC. BSCs must be certified every year.

    31. RYERSON UNIVERSITY 31 During use of a Biological Safety Cabinet: ensure material and aerosol-generating equipment is placed near the back of the hood do not block any vents use techniques that reduce splatter and aerosols general work flow should be from clean to contaminated areas minimize movement so as not to impede air flow Prepare everything you need ahead of time so you arenθt moving in and out of the hood. Rapid and excessive movements in the hood affect air flow. Open flame is not recommended unless there is absolutely no other method to perform what you want (i.e. can micro-burners be used). Contact the EHSS before deciding to use open flame. Prepare everything you need ahead of time so you arenθt moving in and out of the hood. Rapid and excessive movements in the hood affect air flow. Open flame is not recommended unless there is absolutely no other method to perform what you want (i.e. can micro-burners be used). Contact the EHSS before deciding to use open flame.

    32. RYERSON UNIVERSITY 32 After completion of work: Leave blower on at least 5 minutes to purge cabinet Remove and decontaminate equipment and materials Disinfect cabinet surfaces Turn off blower and fluorescent lamp, turn on UV lamp

    33. RYERSON UNIVERSITY 33 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    34. RYERSON UNIVERSITY 34 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    35. RYERSON UNIVERSITY 35 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets

    36. RYERSON UNIVERSITY 36 Maintenance of a Cabinet: Twice daily - Work surfaces wiped down Weekly - UV lamp should be wiped clean Monthly - All vertical surfaces wiped down Annually - certification * UV light is only effective as long as the light is well maintained. Dark spots signify a loss of effectiveness.* UV light is only effective as long as the light is well maintained. Dark spots signify a loss of effectiveness.

    37. RYERSON UNIVERSITY 37 Continue on to: Human Blood and Body Fluids

    38. RYERSON UNIVERSITY 38 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Human Blood and Body Fluids

    39. RYERSON UNIVERSITY 39 RISK OF EXPOSURE DEPENDENT ON: pathogen involved type of body fluid route of exposure duration of exposure volume of blood involved in exposure concentration of virus at time of exposure PPE worn

    40. RYERSON UNIVERSITY 40 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Human Blood and Body Fluids

    41. RYERSON UNIVERSITY 41 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Universal Precautions

    42. RYERSON UNIVERSITY 42 Continue on to: Needle Stick Injury Prevention

    43. RYERSON UNIVERSITY 43 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention

    44. RYERSON UNIVERSITY 44 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention

    45. RYERSON UNIVERSITY 45 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention

    46. RYERSON UNIVERSITY 46 Continue on to: Signs and Labels

    47. RYERSON UNIVERSITY 47

    48. RYERSON UNIVERSITY 48 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels

    49. RYERSON UNIVERSITY 49 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels

    50. RYERSON UNIVERSITY 50 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels

    51. RYERSON UNIVERSITY 51 Continue on to: Personal Protective Equipment

    52. RYERSON UNIVERSITY 52 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Personal Protective Equipment Responsibility of both the user and the supervisor to ensure that PPE is appropriately worn Lab coat (buttoned) Disposable gloves Safety glasses Footwear

    53. RYERSON UNIVERSITY 53 Personal protective equipment (PPE) is only effective if correctly selected, fitted, used and cared for, and the individual is trained Criteria for consideration routes of exposure that need to be blocked degree of protection offered specific to each level of containment Ensure PPE is removed before leaving the lab.

    54. RYERSON UNIVERSITY 54 Lab Coats/Gowns long-sleeved, knee length buttoned or ideally with snaps periodic cleaning required lab coat should be worn in lab area only Closed toed shoes protect against spills and injuries from dropped sharps. Elastic cuffs help prevent spills and contaminationClosed toed shoes protect against spills and injuries from dropped sharps. Elastic cuffs help prevent spills and contamination

    55. RYERSON UNIVERSITY 55 Gloves nitrile & vinyl for work with biological agents avoid latex gloves if possible consider tensile characteristics, length of cuff remove gloves when walking in hallways double glove or do not touch uncontaminated items in lab when wearing gloves

    56. RYERSON UNIVERSITY 56

    57. RYERSON UNIVERSITY 57 Footwear closed toed shoes should always be worn sandals or open toed shoes are not allowed in a biohazard lab Closed toed shoes protect against spills and injuries from dropped sharps. Elastic cuffs help prevent spills and contaminationClosed toed shoes protect against spills and injuries from dropped sharps. Elastic cuffs help prevent spills and contamination

    58. RYERSON UNIVERSITY 58 Continue on to: Purchasing/Transfer

    59. RYERSON UNIVERSITY 59 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Purchasing

    60. RYERSON UNIVERSITY 60 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Transfer

    61. RYERSON UNIVERSITY 61 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Transfer of Human Cells/Tissue

    62. RYERSON UNIVERSITY 62 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Inventory Control

    63. RYERSON UNIVERSITY 63 Continue on to: Decontamination & Waste Disposal

    64. RYERSON UNIVERSITY 64 There is no universal decontamination method for biological materials A spectrum of chemical and physical methods for decontamination exist Environmental and other factors can influence efficacy There is a specific vocabulary for decontamination

    65. RYERSON UNIVERSITY 65 Decontamination Disinfection or sterilization of contaminated surfaces and/or articles to make them suitable for use Disinfectant An agent, usually chemical, that inactivates viruses or kills vegetative microbes but not necessarily resistant forms such as spores Sterilization Act or process (physical or chemical) that destroys or eliminates all forms of life, especially microorganisms

    66. RYERSON UNIVERSITY 66 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Decontamination

    67. RYERSON UNIVERSITY 67 Choice depends on: Type of material to be disinfected Organic load Chemical characteristics Most common are chlorine compounds and alcohols (broad range) Chemical characteristics (corrosion, toxic, flammable, etc.)Chemical characteristics (corrosion, toxic, flammable, etc.)

    68. RYERSON UNIVERSITY 68 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Chemical Decontamination

    69. RYERSON UNIVERSITY 69 Heat: Autoclaving (most practical and recommended) Incineration (for disposal of sharps and tissues) Irradiation: UV light (wavelength of 253 nm is germicidal) Gamma (disrupts DNA and RNA) Filtration HEPA (biological safety cabinets, ventilation) 0.2 micron (physically removes particulates ) Incineration is done of siteIncineration is done of site

    70. RYERSON UNIVERSITY 70 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves

    71. RYERSON UNIVERSITY 71 Items that CAN be autoclaved: cultures and stocks of infectious material culture dishes and related devices discarded live and attenuated vaccines contaminated solid items (petri dishes, eppendorf tips, pipettes, gloves, paper towels) Items that CANNOT be autoclaved: chemical or radioactive waste certain kinds of plastics Material containing solvents, volatile or corrosive chemicals (Phenol, trichloroacetic acid, ether, chloroform) Chemotherapeutic agents.Material containing solvents, volatile or corrosive chemicals (Phenol, trichloroacetic acid, ether, chloroform) Chemotherapeutic agents.

    72. RYERSON UNIVERSITY 72 Items waiting for autoclaving are placed in specially designated biohazard waste containers containing autoclave bags Material containing solvents, volatile or corrosive chemicals (Phenol, trichloroacetic acid, ether, chloroform) Chemotherapeutic agents.Material containing solvents, volatile or corrosive chemicals (Phenol, trichloroacetic acid, ether, chloroform) Chemotherapeutic agents.

    73. RYERSON UNIVERSITY 73 PREPARATION OF WASTE: Use only approved autoclave bags. Do not overfill autoclave bags Separate material for re-use from that which will be disposed and dry from liquid material . If outside of bag is contaminated, double bag. The autoclave bags must not be tightly closed when placed inside the autoclave. Steam must penetrate inside the containers and circulate freely in order for complete sterilization to occur All flasks containing biological material should be capped with aluminum foil. Effectiveness of decontamination from a steam autoclave depends on appropriately loading items into the autoclave. Autoclaving success depends on heat penetrating all material in the bag.Autoclaving success depends on heat penetrating all material in the bag.

    74. RYERSON UNIVERSITY 74 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves

    75. RYERSON UNIVERSITY 75 Treated waste (through autoclaving or other effective method of sterilization) is no longer considered biohazardous and can be disposed in the regular waste stream ALL biological waste must be treated prior to disposal (including level 1 agents).

    76. RYERSON UNIVERSITY 76 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal –External Contractor

    77. RYERSON UNIVERSITY 77 Continue on to: Large Scale Work

    78. RYERSON UNIVERSITY 78 Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Large Scale or High Concentrations

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