4. STANDARD OPERATING PROCEDURES

4. STANDARD OPERATING PROCEDURES
RYERSON UNIVERSITY

4. Standard Operating Procedures
Biological Safety Training – Certificate Holder and User Training BIOLOGICAL SAFETY 4. Standard Operating Procedures 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 RYERSON UNIVERSITY

Good microbiological practice:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control Good microbiological practice: is a basic code of practice to handle biological agents Involves specific sterile techniques of product protection, contamination control applied to all types of work involving microorganisms irrespective of containment level to reduce the risk of exposure and infection Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Good microbiological practices prevent contamination of:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control Good microbiological practices prevent contamination of: laboratory workers the environment experimental samples Good work practices can significantly reduce the risk of: aerosol production contamination of experimental equipment surfaces contains the biological agent reduces the risk of infection Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Before starting any manipulations Before leaving the lab
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control – Hand Washing When to wash? Before starting any manipulations Before leaving the lab When hands are obviously soiled Before and after completing any task in a biosafety cabinet Every time gloves are removed Before contact with one’s face or mouth At the end of the day Rayonnement - Danger - Radiation RYERSON UNIVERSITY

liquid dispensers should be used rather than bars
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control – Hand Washing 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? RYERSON UNIVERSITY

Wet hands with warm water Dispense soap into a cupped hand
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Infection Control – Hand Washing 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. RYERSON UNIVERSITY

Continue on to: Containment of Aerosols
Return to SOP Main Menu RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols Aerosols are gaseous suspensions of fine solid or liquid particles ranging in sizes from 0.01 to 100 µm and can remain suspended in air for extended periods of time. Pathogens such as viruses and bacteria are so small that they can travel within one aerosol droplet and be dispersed by building ventilation. Aerosols can settle on many surfaces where personnel may unwittingly be exposed to a potentially infectious material through the risk of direct contact. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols When appropriate, other primary barriers such as splash shields, face protection or gowns should also be used. When conducting procedures that create considerable aerosols or when using agents classified at Containment Level 2, work should be conducted in a Biological Safety Cabinet Rayonnement - Danger - Radiation RYERSON UNIVERSITY

using centrifuges, shakers, blenders, opening pressurized vessels,
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols The generation and dispersal of aerosols must be minimized and controlled. Numerous procedures and devices which can result in the generation of aerosols: pouring liquids, using centrifuges, shakers, blenders, opening pressurized vessels, inserting a hot loop into a culture, pipetting, etc. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Use a shielded electric incinerator.
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols 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. RYERSON UNIVERSITY

Mouth pipetting is prohibited.
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Containment of Aerosols Mouth pipetting is prohibited. All biohazardous materials should be pipetted in BSC’s. Never force fluids out, use ‘to deliver’ pipettes. To avoid splashes, allow discharge to run down dispense the receiving container wall. Never mix material by suction and expulsion. Reusable pipettes should be placed horizontally in a disinfectant filled pan. Autoclave before reuse. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Continue on to: Working in Containment Level 1 & 2 Areas

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas The following requirements are basic for any laboratory using biological agents. These requirements follow Health Canada’s Laboratory Biosafety Guidelines 3rd ed 2004: Good microbiological laboratory practices intended to avoid the release of infectious agents are to be used. A documented procedural (safety) manual must be available for all staff and its requirements followed. The manual must be reviewed and updated regularly Rayonnement - Danger - Radiation RYERSON UNIVERSITY

eating, drinking, smoking storing food or utensils applying cosmetics
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas PROHIBITED: eating, drinking, smoking storing food or utensils applying cosmetics inserting or removing contact lenses oral pipetting loose long hair - must be tied back or restrained. doors to laboratories must not be left open (does not apply to an open area within a laboratory) Rayonnement - Danger - Radiation RYERSON UNIVERSITY

WORK AREA laboratory must be kept neat, and clean
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas WORK AREA laboratory must be kept neat, and clean storage of materials not pertinent to the work and cannot be easily decontaminated must be minimized paperwork and report writing should be kept separate from biohazardous work areas. access to laboratory and support areas is limited to authorized personnel extreme caution must be used when handling needles and syringes to avoid autoinoculation and the generation of aerosols during use and disposal. Needles must not be bent or sheared. disposable needles and syringes must not be replaced in their cap but placed into a puncture-resistant container Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas WORK AREA technical procedures must be performed in a manner that minimizes the creation of aerosols work surfaces must be cleaned and decontaminated with the appropriate disinfectant at the end of the day and after any spill of potentially hazardous material. loose or cracked work surfaces must be repaired or replaced. contaminated materials and equipment leaving the laboratory for servicing or disposal must be appropriately decontaminated and labelled or tagged as contaminated Rayonnement - Danger - Radiation RYERSON UNIVERSITY

PERSONAL PROTECTIVE EQUIPMENT
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas PERSONAL PROTECTIVE EQUIPMENT laboratory coats must be worn and fastened by all personnel working in the laboratory. suitable footwear with closed toes and heels and preferably with non-slip soles must be worn in laboratory protective laboratory clothing must not be worn in non-laboratory areas; lab coats must not be stored in contact with street clothing suspected contaminated clothing must be decontaminated before laundering eye protection must be worn when necessary to protect the eyes hands must be washed after gloves are removed, before leaving the laboratory, and after handling materials known or suspected to be contaminated, even when gloves have been worn Rayonnement - Danger - Radiation RYERSON UNIVERSITY

retraining programs should also be implemented.
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas TRAINING: Lab personnel must receive training on the potential hazards and the necessary precautions to prevent exposure to infectious agents and release of contained material personnel must show evidence that they understood the training provided training must be documented and signed by both the employee and supervisor retraining programs should also be implemented. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

DISINFECTION & WASTE DISPOSAL
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas DISINFECTION & WASTE DISPOSAL Disinfectants effective against the agents in use must be available at all times within the areas where the biohazardous material is handled or stored effective concentrations and contact times must be used all contaminate materials must be decontaminated before disposal or reuse efficacy monitoring of autoclaves used for decontamination with biological indicators must be done regularly (i.e. consider weekly, depending on the frequency of use of the autoclave) records of these results and cycle logs (temperature, pressure) must also be kept on file Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 1 areas ACCIDENTS All spills, accidents (needlesticks, punctures, cuts, etc.) and overt or potential exposures must be reported in writing to the laboratory supervisor as soon as circumstances permit. must file two reports: Ryerson University’s Internal Accident Investigation Report filed with the Centre for Environmental Health and Safety Management (CEHSM) within 24 hours of incident Workers’ Compensation Form (WSIB Form 7) form must be sent to Human Resources with 24 hours of incident. Appropriate medical evaluation, surveillance, and treatment must be sought and provided as required. Actions taken to prevent future occurrences should be documented. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

appropriate door sign must be posted outside each laboratory
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Working in CL 2 areas In addition to the safety precautions outlined for Containment Level 1 laboratory additional operational practices are outline below: biological safety cabinets must be used for procedures that may produce infectious aerosols and that involve high concentrations or large volumes of biohazardous material. appropriate door sign must be posted outside each laboratory Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Continue on to: Biological Safety Cabinets

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Biological Safety Cabinets (BSC) provide an effective means of physical containment for biological agents, especially when aerosols are generated. The main role is to provide protection to personnel, and the environment and product. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Protection is achieved through the control of air movement within and prior to leaving the cabinet, and through the use of HEPA (high-efficiency particulate air) filtration HEPA filters are designed to remove particles with a minimum size of 0.3 microns with an efficiency of 99.97%. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures 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 (non-turbulent) which is directed out of the cabinet or down into intakes these cabinets provide product protection only and must not be used when working with any form of biohazard sometimes referred to as Clean Benches. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Cabinet Certification
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Cabinet Certification BSCs must be certified annually or after cabinet has been moved laminar flow hoods should be certified every second year undertaken to ensure the HEPA filter has not be damaged, leaking or plugged. cabinet should be decontaminated prior to any certification activity. HEPA filters & cabinets must be decontaminated prior to disposal Rayonnement - Danger - Radiation RYERSON UNIVERSITY

The effectiveness of a BSC is dependent upon:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets The effectiveness of a BSC is dependent upon: the integrity of the cabinet – (the risk of exposure increases if the integrity is jeopardized) location of cabinet in a room proper microbiological technique and work practices that do not disturb established airflow velocity and cause reverse currents that can re-introduce contaminants into the work area continued maintenance and certification. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Before using the cabinet:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets 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. RYERSON UNIVERSITY

During use of a Biological Safety Cabinet:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets 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 arenèt 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. RYERSON UNIVERSITY

After completion of work:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets 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 RYERSON UNIVERSITY

Flames in Biological Safety Cabinets
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Flames in Biological Safety Cabinets The use of flame to sterilize equipment within a BSC is controversial. An open flame in a BSC creates several major problems: Flame creates turbulence, which disrupts the pattern of air supplied to the work surface, therefore reducing maximum efficiency. Heat build-up inside the BSC that may damage the HEPA filters Creates a fire hazard. It is recommended that flame should not be used in BSC's. Alternatives to flames include using disposable sterile inoculating loops and needles, or pre autoclaved equipment in sterile packaging. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Flames in Biological Safety Cabinets
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Flames in Biological Safety Cabinets If a flame is required, a touch-plate micro-burner equipped with a pilot light to provide a flame on demand may be used. This device will minimize internal cabinet air disturbances and heat build-up. During use, the heat source should be placed to the rear of the workspace where resulting air turbulence will have minimal effect. An emergency shut off valve should be placed just outside the BSC gas supply line and during the use of any burner, all combustible materials and solvents must be removed. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets Ultraviolet 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 the terminal ends are blackened even slightly UV lamps themselves are potential hazards since UV light can be harmful to the eyes and skin and should therefore be turned off when work is being conducted in the cabinet. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Maintenance of a Cabinet:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Biological Safety Cabinets 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. RYERSON UNIVERSITY

Continue on to: Human Blood and Body Fluids

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Human Blood and Body Fluids blood borne pathogens, which are microorganisms that are present in blood and bodily fluids and are capable of causing disease in exposed individuals exposure to human blood and bodily fluids (e.g., semen, cerebrospinal, amniotic,), tissue cultures, or organ cultures increases the risk pathogens of greatest concern are hepatitis B virus, hepatitis C virus and the Human Immunodefiency Virus (HIV) Rayonnement - Danger - Radiation RYERSON UNIVERSITY

RISK OF EXPOSURE DEPENDENT ON: pathogen involved type of body fluid
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Human Blood and Body Fluids 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 RYERSON UNIVERSITY

UNIVERSAL PRECAUTIONS:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Human Blood and Body Fluids UNIVERSAL PRECAUTIONS: The minimum standard of practice to prevent exposure to blood borne pathogens and includes: education personal protective equipment hand washing safe work practices Universal precautions assumes the material to be infectious and required to use control measures to prevent skin and mucous membrane exposure. These precautions must always be used when handling blood or body fluids. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Gloves should be worn when handling potentially contaminated surfaces.
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Universal Precautions Gloves should be worn when handling potentially contaminated surfaces. Avoid touching items that are NOT contaminated when gloves are being worn. PPE should be worn during procedures that are likely to generate droplets of blood or bodily fluids. Contaminated lab coats and gloves should be removed immediately after procedure is completed. 5. Hand washing is the most important preventative tool - hands should be washed immediately after gloves are removed and before leaving a work area. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Continue on to: Needle Stick Injury Prevention

Injuries from sharps (needles, syringes, etc.) may result from:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention Injuries from sharps (needles, syringes, etc.) may result from: lack of training on proper work practices crowded work conditions incorrect recapping of non retracting needles poor disposal practices [risk of injury exists not only to the user, but also support staff such as caretaking or hazardous waste disposal personnel] Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention Should a needle stick injury occur it is important to report it immediately and seek a medical assessment. Such injuries must not be ignored - potential laboratory associated infections could develop without appropriate treatment. Rayonnement - Danger - Radiation RYERSON UNIVERSITY

To avoid needle stick injury use:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Needle Stick Injury Prevention To avoid needle stick injury use: needle-less techniques where possible retractable sharps fill syringes carefully do not bend, or recap needles approved designated sharps containers should be used for disposal that will not allow penetration or direct access to sharps Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Continue on to: Signs and Labels

Biohazardous or infectious materials fall under;
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels Biohazardous or infectious materials fall under; Class D, division 3 of WHMIS (Poisonous and Infectious Material - Biohazardous Infectious Material) Class 6.2 of Transport Canada Transport of Dangerous Goods Act (Infectious Substances) RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels ENTRANCES Biohazard signs must be posted on doors to rooms where biohazardous materials are used (Level 2 or greater) Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels Biohazard labels must also be placed on containers, equipment and storage units (e.g., fridges, freezers) used for biological agents. RYERSON UNIVERSITY

INSIDE LAB Internal Ryerson Biosafety Certificate
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Signs & Labels INSIDE LAB (posted prominently) Internal Ryerson Biosafety Certificate Current list of authorized users RYERSON UNIVERSITY

Continue on to: Personal Protective Equipment

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

Criteria for consideration
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures 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. RYERSON UNIVERSITY

long-sleeved, knee length buttoned or ideally with snaps
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Personal Protective Equipment 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 contamination RYERSON UNIVERSITY

nitrile & vinyl for work with biological agents
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Personal Protective Equipment 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 RYERSON UNIVERSITY

safety glasses or goggles to protect the eyes
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Personal Protective Equipment Safety Glasses safety glasses or goggles to protect the eyes RYERSON UNIVERSITY

NO Footwear closed toed shoes should always be worn
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Personal Protective Equipment 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 contamination NO RYERSON UNIVERSITY

Continue on to: Purchasing/Transfer

Records of all purchases must be maintained for inspection
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Purchasing ALL PURCHASES OF CL 2 BIO AGENTS MUST BE APPROVED BY BSO PRIOR TO PURCHASE Only Certificate Holders or designated authorized users may purchase biological agents Import permits required from United States for certain animal or human pathogens Records of all purchases must be maintained for inspection RYERSON UNIVERSITY

prior BSO approval to ensure appropriate containment available
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Transfer WITHIN UNIVERSITY prior BSO approval to ensure appropriate containment available unauthorized lending or borrowing of biohazards not permitted between labs Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Transfer of Human Cells/Tissue OUTSIDE UNIVERSITY prior BSO approval for lending or borrowing of biohazards from other institutions before arrival on campus valid Research Ethics Board (REB) approval is required prior to the transfer of material receiving institution must have appropriate containment materials must be properly packaged and documentation inventory must be adjusted if material is being received by Ryerson, then a copy of the other institution’s Research Ethic Board approval is also required Online applications for Research Ethic Board approval is available at: RYERSON UNIVERSITY

Changes require a revision to the Biosafety Certificate
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Inventory Control Certificate Holder required to use Inventory control Form for all biological agents Inventory records must be kept up to date and available to the BSO for inspection Types of biological organisms should be kept current with the list noted in the Biosafety Certificate Changes require a revision to the Biosafety Certificate Rayonnement - Danger - Radiation RYERSON UNIVERSITY

Continue on to: Decontamination & Waste Disposal

There is no universal decontamination method for biological materials
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures 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 RYERSON UNIVERSITY

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

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Decontamination laboratory bench tops, other surfaces, and all equipment contaminated with biohazards must be decontaminated prior to reuse, servicing, transfer or disposal. procedures must be in place to ensure the effectiveness of the methods used to decontaminate items. RYERSON UNIVERSITY

Most common are chlorine compounds and alcohols (broad range)
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Chemical Decontamination 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.) RYERSON UNIVERSITY

Chemicals include: 2% -10% domestic bleach (hypochorite solution)
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Chemical Decontamination Chemicals include: 2% -10% domestic bleach (hypochorite solution) 70% Ethanol Quaternary ammonia 6% formulated Hydrogen peroxide 10% formalin 2% glutaraldeyhye phenolic compounds RYERSON UNIVERSITY

Heat: Irradiation: Filtration
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Physical Decontamination 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 site RYERSON UNIVERSITY

sterilization is achieved through autoclaves
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves sterilization is a process which results in the total destruction of all living and viable organisms (with a probability of 1 in 1 million that 1 organism survived) sterilization is achieved through autoclaves autoclaves utilize both pressure and high temperatures to provide an effective way of sterilizing items RYERSON UNIVERSITY

Items that CAN be autoclaved:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves 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. RYERSON UNIVERSITY

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

Use only approved autoclave bags. Do not overfill autoclave bags
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves 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. RYERSON UNIVERSITY

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal -Autoclaves after materials have been autoclaved, the bags must be tightly sealed and placed inside unmarked green or black regular waste garbage bags garbage bags are then sealed by laboratory personnel and placed in a designated area away from the biohazard area for regular garbage disposal by caretaking staff Disposal RYERSON UNIVERSITY

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

Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Waste Disposal –External Contractor Any waste that cannot be treated (i.e. sharps, tissues) remains biohazardous waste and must be disposed off site using a licensed waste disposal contractor. Contaminated materials destined for disposal must be placed in clearly marked red or orange biohazard bags and sealed. If using an external contractor to dispose of non autoclaved waste materials, please contact the Biological Safety Officer to assist in registration of waste materials with the Ontario Ministry of the Environment RYERSON UNIVERSITY

Continue on to: Large Scale Work

Containment levels based on laboratory scale operations:
Biological Safety Training – Certificate Holder and User Training 4. Standard Operating Procedures Large Scale or High Concentrations Containment levels based on laboratory scale operations: increase in containment may be required if working with high concentrations or shifting to manipulations involving >10 litres increase in the amount of potentially infectious material may require additional safety precautions and/or increase in containment risk assessment should determine whether there is an increase in aerosol generation RYERSON UNIVERSITY

4. STANDARD OPERATING PROCEDURES

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