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Biological Safety CabinetsFlames in Biological Safety Cabinets If a flame is required, a touch-plate micro-burner equipped with a pilotlight to provide a flame on demand may be used.This device will minimize internal cabinet air disturbances and heatbuild-up.During use, the heat source should be placed to the rear of theworkspace where resulting air turbulence will have minimal effect.An emergency shut off valve should be placed just outside the BSCgas supply line and during the use of any burner, all combustiblematerials and solvents must be removed.

Biological Safety CabinetsUltraviolet Lamps: Intended to destroy microorganisms in air or on exposed surfaces.Have limited penetrating power and only effective when properly cleaned, maintained.Dust is attracted to the lamps which reduces the transmission of the germicidal effect.Have a limited life span – even if the blue-violet glows, the lamps are not effective if theterminal ends are blackened even slightly.UV lamps themselves are potential hazards since UV light can be harmful to the eyesand skin and should therefore be turned off when work is being conducted in thecabinet.

Biological Safety CabinetsMaintenance of a Cabinet: twice daily: work surfaces wiped downweekly: UV lamp should be wiped cleanmonthly: all vertical surfaces wiped downannually: certification

Human Blood and BodyFluids

Human Blood and Body Fluids blood borne pathogens, which are microorganisms that are present in bloodand bodily fluids and are capable of causing disease in exposed individuals exposure to human blood and bodily fluids (e.g., semen, cerebrospinal,amniotic, etc.), tissue cultures, or organ cultures increases the risk pathogens of greatest concern are hepatitis B virus, hepatitis C virus and theHuman Immunodeficiency Virus (HIV)

Human Blood and Body FluidsRisk 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

Human Blood and Body FluidsUniversal precautionsThe minimum standard of practice to prevent exposure to bloodborne pathogens, including: education personal protective equipment hand washing safe work practicesUniversal precautions assumes the material to be infectious and required to use controlmeasures to prevent skin and mucous membrane exposure.These precautions must always be used when handling blood or body fluids.

Universal Precautions1.Gloves should be worn when handling potentially contaminated surfaces.2.Avoid touching items that are NOT contaminated when gloves are being worn.3.PPE should be worn during procedures that are likely to generate droplets of blood orbodily fluids.4.Contaminated lab coats and gloves should be removed immediately after procedure iscompleted.5.Hand washing is the most important preventative tool: hands should be washedimmediately after gloves are removed and before leaving a work area.

Needle Stick InjuryPrevention

Needle Stick Injury PreventionInjuries from sharps (needles, syringes, etc.) may result from: lack of training on proper work practicescrowded work conditionsincorrect recapping of non retracting needlespoor disposal practicesRisk of injury exists not only to the user, but also support staff such ascaretaking or hazardous waste disposal personnel.

Needle Stick Injury PreventionShould a needle stick injury occur it is important toreport it immediately and seek a medicalassessment.Such injuries must not be ignored - potentiallaboratory associated infections could developwithout appropriate treatment.

Needle Stick Injury PreventionTo avoid needle stick injury: use needle-less techniques where possibleuse retractable sharpsfill syringes carefullydo not bend, or recap needlesuse approved designated sharps containers should be used for disposal thatwill not allow penetration or direct access to sharps

Signs and Labels

Signs and LabelsBiohazardous or infectious materials fall under:Biohazardous Infectious Material of WHMISClass 6.2 of Transport Canada’s Transportation ofDangerous Goods Act (Infectious Substances)

Signs and LabelsEntrancesBiohazard signs must be posted on doorsto rooms where biohazardous materialsare used (Level 2 or greater)

Signs and LabelsBiohazard labels must also be placed on containers, equipment andstorage units (e.g., fridges, freezers) used for biological agents.

Signs and LabelsInside lab(posted prominently) Internal Ryerson Biosafety PermitCurrent list of authorized users

Personal ProtectiveEquipment

Personal Protective Equipment lab coat (buttoned)disposable glovessafety glassesfootwear (closed toe)Responsibility of both the user and the supervisor to ensure that PPEis appropriately worn and kept in good condition

Personal Protective Equipment 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.

Personal Protective EquipmentLab Coats/Gowns long-sleeved, knee lengthbuttoned or ideally with snapsperiodic cleaning requiredlab coat should be worn in lab area only

Personal Protective EquipmentGloves nitrile and vinyl for work with biological agentsavoid latex gloves if possibleconsider tensile characteristics, length of cuffremove gloves when walking in hallwaysdouble glove or do not touch uncontaminated items in labwhen wearing gloves

Personal Protective EquipmentSafety Glasses safety glasses or goggles to protect the eyes

Personal Protective EquipmentFootwear closed toed shoes should always be wornsandals or open toed shoes are not allowed in a biohazard labNO

Purchasing/Transfer

Purchasing ALL PURCHASES OF CL 2 BIO AGENTS MUST BEAPPROVED BY BSO PRIOR TO PURCHASE. Only Certificate Holders or designated authorized users maypurchase biological agents. Import permits required from United States for certain animalor human pathogens. Records of all purchases must be maintained for inspection.

TransferWithin the university prior BSO approval to ensure appropriate containment available unauthorized lending or borrowing of biohazards not permitted between labs

Transfer of Human Cells/TissueOutside the university prior BSO approval for lending or borrowing of biohazards fromother institutions before arrival on campusreceiving institution must have appropriate containmentmaterials must be properly packaged and documentationpersons preparing and/or transporting package must have validTDG Class 6 Training certificateinventory must be adjusted

Inventory ControlPermit Holder must have up to date inventory list of agents onpermit, which will be used by the BSO for inspection. any changes requires a revision to the biosafety permit.find a link on the EHS’ Biosafety webpage

Decontamination & WasteDisposal

Decontamination 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

Decontamination VocabularyDecontamination: Disinfection or sterilization of contaminatedsurfaces and/or articles to make them suitable for useSterilization: Act or process (physical or chemical) that destroys oreliminates all forms of life, especially microorganismsDisinfectant: An agent, usually chemical, that inactivates virusesor kills vegetative microbes but not necessarily resistant forms suchas spores

Decontamination laboratory bench tops, other surfaces, and all equipmentcontaminated with biohazards must be decontaminated priorto reuse, servicing, transfer or disposal procedures must be in place to ensure the effectiveness of themethods used to decontaminate items

Chemical DecontaminationChoice depends on: type of material to be disinfectedorganic loadchemical characteristicsMost common are chlorine compounds and alcohols (broad range)

Chemical DecontaminationChemicals include: 2% -10% domestic bleach (hypochlorite solution) 70% Ethanol quaternary ammonia 6% formulated Hydrogen peroxide 10% formalin 2% glutaraldehyde phenolic compounds

Physical DecontaminationHeat: 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 )

Sterilization by Autoclaves sterilization is a process which results inthe total destruction of all living and viableorganisms (with a probability of 1 in 1million that 1 organism survived)sterilization is achieved throughautoclavesautoclaves utilize both pressure and hightemperatures to provide an effective wayof sterilizing items

Waste Disposal Ryerson disposes of its biological wastes though a certifiedbiomedical waste disposal company. Such companies collect properly packaged biological wastefor industrial, off-site autoclaving and/or incineration

Waste DisposalIf you currently have solid waste:1. Set up the supplied cardboard boxes with yellow plastic linerand place your bag inside (you can put multiple bags until theyellow liner/box is full)2. Tie up liner, close and tape shut the box3. If your waste contains cytotoxic drugs (i.e., chemo agents), puta “cytotoxic” label on the box4. With a marker, write lab name and date on side of box (e.g.,Smith lab/ Jan 12, 2017)

Solid Waste in Cardboard Boxes

Waste Disposal SuppliesColour CodingYellow for regular biological wasteRed for cytotoxic waste Any material that may have come into contact with acytotoxic drug (such as chemotherapeutic agents) whichinhibit or prevent the function of cells.

Waste Disposal SuppliesPails Use pails to collect waste that could puncture through plasticbags, such as serological pipettes, pipette tips, glass tubes,slides, etc. Use pails to collect liquids: do NOT pour liquids directly intopails, keep liquids in small bottles, then tighten their lids andplace in pails

Waste Disposal SuppliesBoxes with yellow liners For solid waste that cannot puncture a plastic bag (e.g., contaminated gloves,paper towels, unbreakable plastic bottles, etc.) For filled sharps containers

Waste Disposal Supplies Each department will have a location where you can pick up these suppliesー Current locations for Chemistry & Biology department – KHN-202D Call main department to coordinateー Other departments will need to find a common location for supplies orcontact EHSYou can also email ehs@ryerson.ca for more supplies if your departmenthas run out

Waste Disposal Process When boxes are full, tie up liner and tape box closedWhen pails are full, close lid tightly until it clicks (locked –cannot be opened again)Label side of boxes and white lid of pails with Lab name andcurrent date Example: Smith lab / Jan 12

Waste Disposal Process Biological waste pick-up will take place every Monday morning Stay current by checking the EHS’ Biosafety webpage for any updates orchangesEach department will have a location where biological waste can be dropped off Current locations KHN-202D Call Chemistry & Biology department to coordinate

Large Scale Work

Large Scale or High ConcentrationsContainment levels based on laboratory scale operations (inaddition to pathogenicity of agents): increase in containment may be required if working with highconcentrations or shifting to manipulations involving 10 litres increase in the amount of potentially infectious material mayrequire additional safety precautions and/or increase incontainment risk assessment should determine whether there is an increasein aerosol generation

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Laminar Flow Cabinets Laminar flow cabinets (LFC) are similar in appearance, but are not Biological Safety Cabinets: only protect the product intake room air which is passed through a pre-filter and a HEPA filter to remove contaminants, dust and other particles purified air then enters the work surface in a laminar flow