RYERSON UNIVERSITY 1 4. STANDARD OPERATING PROCEDURES.

RYERSON UNIVERSITYRYERSON UNIVERSITY 11

4. STANDARD OPERATING 4. STANDARD OPERATING PROCEDURESPROCEDURES


Biological Safety Training – Certificate Holder and User Training Biological Safety Training – Certificate Holder and User Training

BIOLOGICAL SAFETYBIOLOGICAL SAFETY

Infection ControlInfection Control

Containment of AerosolsContainment of Aerosols

Working in CL 1 areasWorking in CL 1 areas

Working in CL 2 areasWorking in CL 2 areas

Biological Safety Biological Safety CabinetsCabinets

Human Blood and Body Human Blood and Body FluidsFluids

Needle Stick Injury Needle Stick Injury PreventionPrevention

4. Standard Operating Procedures

Signs and LabelsSigns and Labels

Personal Protective Personal Protective EquipmentEquipment

Purchasing/TransferPurchasing/Transfer

Inventory ControlInventory Control

Decontamination/Waste Decontamination/Waste DisposalDisposal

Large Scale WorkLarge Scale Work



4. 4. Standard Operating ProceduresStandard Operating ProceduresInfection ControlInfection Control

Rayonnement - Danger - Radiation

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



4. 4. Standard Operating ProceduresStandard Operating ProceduresInfection ControlInfection Control


Good microbiological practices prevent contamination of:

laboratory workersthe environmentexperimental samples

Good work practices can significantly reduce the risk of:aerosol productioncontamination of experimental equipment surfacescontains the biological agent reduces the risk of infection



4. 4. Standard Operating ProceduresStandard Operating ProceduresInfection Control – Hand WashingInfection Control – Hand Washing


When to wash?When to wash?

Before starting any manipulationsBefore starting any manipulations

Before leaving the labBefore leaving the lab

When hands are obviously soiledWhen hands are obviously soiled

Before and after completing any task in a Before and after completing any task in a biosafety cabinetbiosafety cabinet

Every time gloves are removedEvery time gloves are removed

Before contact with one’s face or mouthBefore contact with one’s face or mouth

At the end of the dayAt the end of the day


Frequent hand washing has proven to be the Frequent hand washing has proven to be the single most effective means of avoiding infection single most effective means of avoiding infection ifif done properly and frequentlydone properly and frequently

liquid dispensers should be used rather than barsliquid dispensers should be used rather than bars




Wet hands with warm waterWet hands with warm water

Dispense soap into a cupped handDispense soap into a cupped hand

Spread around hands and between fingersSpread around hands and between fingers

Wash hands for Wash hands for at leastat least 10 sec. 10 sec.

Rinse thoroughly under warm water.Rinse thoroughly under warm water.

Dry hands thoroughly with paper towels.Dry hands thoroughly with paper towels.




Continue on to:Continue on to:Containment of AerosolsContainment of Aerosols

Return to SOP Main MenuReturn to SOP Main Menu



4. 4. Standard Operating ProceduresStandard Operating ProceduresContainment of AerosolsContainment 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.





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





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.


Sterilization of inoculation loops in an open flame Sterilization of inoculation loops in an open flame may create aerosols which may contain viable may create aerosols which may contain viable microorganisms.microorganisms.

Use a shielded electric incinerator.Use a shielded electric incinerator.

Shorter handles minimize vibrations.Shorter handles minimize vibrations.

Disposable plastic loops are good alternatives.Disposable plastic loops are good alternatives.







Mouth pipetting is prohibited.Mouth pipetting is prohibited.

All biohazardous materials should be pipetted in BSC’s.All biohazardous materials should be pipetted in BSC’s.

Never force fluids out, use ‘to deliver’ pipettes.Never force fluids out, use ‘to deliver’ pipettes.

To avoid splashes, allow discharge to run down To avoid splashes, allow discharge to run down dispense the receiving container wall.dispense the receiving container wall.

Never mix material by suction and expulsion.Never mix material by suction and expulsion.

Reusable pipettes should be placed horizontally in a Reusable pipettes should be placed horizontally in a disinfectant filled pan. Autoclave before reuse.disinfectant filled pan. Autoclave before reuse.


Continue on to:Continue on to:Working in Containment Working in Containment

Level 1 & 2 AreasLevel 1 & 2 Areas




4. 4. Standard Operating ProceduresStandard Operating ProceduresWorking in CL 1 areasWorking 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





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)





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





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





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





TRAINING:

Lab personnel must receive training on the potential hazards and the necessary precautions to prevent exposure to infectious agents and release of contained materialpersonnel must show evidence that they understood the training providedtraining must be documented and signed by both the employee and supervisor retraining programs should also be implemented.





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 storedeffective concentrations and contact times must be used all contaminate materials must be decontaminated before disposal or reuseefficacy 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





ACCIDENTSAll 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.





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


Continue on to:Continue on to:Biological Safety CabinetsBiological Safety Cabinets




4. 4. Standard Operating ProceduresStandard Operating Procedures

Biological Safety CabinetsBiological 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.



4. 4. Standard Operating ProceduresStandard Operating Procedures

Biological Safety CabinetsBiological 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%.



4. 4. Standard Operating ProceduresStandard Operating ProceduresLaminar Flow CabinetsLaminar 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.



4. 4. Standard Operating ProceduresStandard Operating ProceduresBiological Safety CabinetsBiological 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





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.


Before using the cabinetBefore using the cabinet::

Ensure BSC is certifiedEnsure BSC is certified

Disinfect work surfaces with Disinfect work surfaces with disinfectantdisinfectant

Place essential items inside Place essential items inside cabinetcabinet

Allow the blower to run for 5-10 Allow the blower to run for 5-10 min before workmin before work




During use of a Biological Safety CabinetDuring use of a Biological Safety Cabinet::

ensure material and aerosol-generating ensure material and aerosol-generating equipment is placed near the back of the hood equipment is placed near the back of the hood

do not block any ventsdo not block any vents

use techniques that reduce splatter and aerosolsuse techniques that reduce splatter and aerosols

general work flow should be from clean to general work flow should be from clean to contaminated areascontaminated areas

minimize movement so as not to impede air flowminimize movement so as not to impede air flow




After completion of workAfter completion of work::

Leave blower on at least 5 Leave blower on at least 5 minutes to purge cabinetminutes to purge cabinet

Remove and decontaminate Remove and decontaminate equipment and materialsequipment and materials

Disinfect cabinet surfacesDisinfect cabinet surfaces

Turn off blower and fluorescent Turn off blower and fluorescent lamp, turn on UV lamplamp, turn on UV lamp







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 filtersCreates 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.





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.





Ultraviolet Lamps:

Intended to destroy microorganisms in air or on exposed surfacesHave limited penetrating power and only effective when properly

cleaned, maintainedDust is attracted to the lamps which reduces the transmission of the

germicidal effectHave 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.


Maintenance of a CabinetMaintenance of a Cabinet::

Twice daily - Work surfaces wiped downTwice daily - Work surfaces wiped down

Weekly - UV lamp should be wiped cleanWeekly - UV lamp should be wiped clean

Monthly - All vertical surfaces wiped downMonthly - All vertical surfaces wiped down

Annually - certificationAnnually - certification




Continue on to:Continue on to:Human Blood and Body FluidsHuman Blood and Body Fluids




4. 4. Standard Operating ProceduresStandard Operating ProceduresHuman Blood and Body FluidsHuman 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)


RISK OF EXPOSURE DEPENDENT ONRISK OF EXPOSURE DEPENDENT ON::

pathogen involvedpathogen involved

type of body fluidtype of body fluid

route of exposureroute of exposure

duration of exposureduration of exposure

volume of blood involved in exposurevolume of blood involved in exposure

concentration of virus at time of exposureconcentration of virus at time of exposure

PPE wornPPE worn







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.



4. 4. Standard Operating ProceduresStandard Operating ProceduresUniversal PrecautionsUniversal Precautions


1. 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 or bodily fluids.

4. 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.


Continue on to:Continue on to:Needle Stick Injury PreventionNeedle Stick Injury Prevention




4. 4. Standard Operating ProceduresStandard Operating ProceduresNeedle Stick Injury PreventionNeedle 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]





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.





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


Continue on to:Continue on to:Signs and LabelsSigns and 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)


4. 4. Standard Operating ProceduresStandard Operating ProceduresSigns & LabelsSigns & Labels


Biological Safety Training – Certificate Holder and User TrainingBiological Safety Training – Certificate Holder and User Training

4. 4. Standard Operating ProceduresStandard Operating Procedures Signs & LabelsSigns & Labels

ENTRANCES

Biohazard signs must be posted Biohazard signs must be posted on doors to roomson doors to rooms where biohazardous materials are used (Level 2 or where biohazardous materials are used (Level 2 or greater)greater)




4. 4. Standard Operating ProceduresStandard Operating ProceduresSigns & LabelsSigns & Labels

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



4. 4. Standard Operating ProceduresStandard Operating Procedures Signs & LabelsSigns & Labels

INSIDE LAB (posted prominently)

Internal Ryerson Biosafety Certificate Current list of authorized users


Continue on to:Continue on to:Personal Protective EquipmentPersonal Protective Equipment




4. 4. Standard Operating ProceduresStandard Operating ProceduresPersonal Protective EquipmentPersonal Protective Equipment

Responsibility of both the user and the supervisor to Responsibility of both the user and the supervisor to ensure that PPE is appropriately wornensure that PPE is appropriately worn

• Lab coat (buttoned)Lab coat (buttoned)• Disposable glovesDisposable gloves• Safety glassesSafety glasses• FootwearFootwear


Personal protective equipment (PPE) is only effective if Personal protective equipment (PPE) is only effective if correctly selected, fitted, used and cared for, and the correctly selected, fitted, used and cared for, and the individual is trainedindividual is trained

Criteria for considerationCriteria for consideration• routes of exposure that need to be blockedroutes of exposure that need to be blocked• degree of protection offereddegree of protection offered• specific to each level of containmentspecific to each level of containment

Ensure PPE is removed before leaving the lab.Ensure PPE is removed before leaving the lab.




Lab Coats/GownsLab Coats/Gowns

long-sleeved, knee length long-sleeved, knee length

buttoned or ideally with snapsbuttoned or ideally with snaps

periodic cleaning requiredperiodic cleaning required

lab coat should be worn in lab area lab coat should be worn in lab area onlyonly




GlovesGloves

nitrile & vinyl for work with biological agentsnitrile & vinyl for work with biological agents

avoid latex gloves if possibleavoid latex gloves if possible

consider tensile characteristics, length of cuffconsider tensile characteristics, length of cuff

remove gloves when walking in hallways remove gloves when walking in hallways

double glove or do not touch uncontaminated double glove or do not touch uncontaminated items in lab when wearing glovesitems in lab when wearing gloves




Safety GlassesSafety Glasses

safety glasses or goggles to protect the eyessafety glasses or goggles to protect the eyes




FootwearFootwear

closed toed shoes should always be wornclosed toed shoes should always be worn

sandals or open toed shoes are not allowed in a sandals or open toed shoes are not allowed in a biohazard lab biohazard lab




Continue on to:Continue on to:Purchasing/TransferPurchasing/Transfer




4. 4. Standard Operating ProceduresStandard Operating Procedures PurchasingPurchasing

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 Import permits required from United States for certain animal or human pathogensor human pathogens

Records of all purchases must be maintained for inspectionRecords of all purchases must be maintained for inspection



4. 4. Standard Operating ProceduresStandard Operating Procedures TransferTransfer

WITHIN UNIVERSITY

prior BSO approval to ensure appropriate containment available

unauthorized lending or borrowing of biohazards not permitted between labs




4. 4. Standard Operating ProceduresStandard Operating Procedures Transfer of Human Cells/TissueTransfer of Human Cells/Tissue

Online applications for Research Ethic Board approval is available at:http://www.ryerson.ca/about/vpresearch/ethics.html

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 containmentmaterials must be properly packaged and documentationinventory must be adjustedif material is being received by Ryerson, then a copy of the other

institution’s Research Ethic Board approval is also required


Biological Safety Training Biological Safety Training – Certificate Holder and User Training – Certificate Holder and User Training

4. 4. Standard Operating ProceduresStandard Operating Procedures Inventory ControlInventory 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



Continue on to:Continue on to:Decontamination & Waste Decontamination & Waste

DisposalDisposal



There is no universal decontamination method There is no universal decontamination method for biological materialsfor biological materials

A spectrum of chemical and physical methods A spectrum of chemical and physical methods for decontamination existfor decontamination exist

Environmental and other factors can influence Environmental and other factors can influence efficacyefficacy

There is a specific vocabulary for There is a specific vocabulary for decontaminationdecontamination


4. 4. Standard Operating ProceduresStandard Operating Procedures DecontaminationDecontamination


• DecontaminationDecontamination• Disinfection or sterilization of contaminated surfaces Disinfection or sterilization of contaminated surfaces

and/or articles to make them suitable for useand/or articles to make them suitable for use

• DisinfectantDisinfectant• An agent, usually chemical, that inactivates viruses or An agent, usually chemical, that inactivates viruses or

kills vegetative microbes but not necessarily resistant kills vegetative microbes but not necessarily resistant forms such as sporesforms such as spores

• SterilizationSterilization• Act or process (physical or chemical) that destroys or Act or process (physical or chemical) that destroys or

eliminates all forms of life, especially microorganismseliminates all forms of life, especially microorganisms


4. 4. Standard Operating ProceduresStandard Operating Procedures Decontamination VocabularyDecontamination Vocabulary



4. 4. Standard Operating ProceduresStandard Operating Procedures DecontaminationDecontamination

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.


Choice depends on:Choice depends on:• Type of material to be disinfectedType of material to be disinfected• Organic loadOrganic load• Chemical characteristicsChemical characteristics

Most common are chlorine Most common are chlorine compounds and alcohols (broad compounds and alcohols (broad range)range)


4. 4. Standard Operating ProceduresStandard Operating Procedures Chemical DecontaminationChemical Decontamination



4. 4. Standard Operating ProceduresStandard Operating Procedures Chemical Decontamination Chemical Decontamination

•Chemicals include:

2% -10% domestic bleach (hypochorite solution)2% -10% domestic bleach (hypochorite solution) 70% Ethanol70% Ethanol Quaternary ammoniaQuaternary ammonia 6% formulated Hydrogen peroxide6% formulated Hydrogen peroxide 10% formalin 2% glutaraldeyhye phenolic compounds


Heat:Heat:• Autoclaving (most practical and recommended)Autoclaving (most practical and recommended)• Incineration (for disposal of sharps and tissues)Incineration (for disposal of sharps and tissues)

IrradiationIrradiation::• UV light (wavelength of 253 nm is germicidal)UV light (wavelength of 253 nm is germicidal)• Gamma (disrupts DNA and RNA)Gamma (disrupts DNA and RNA)

FiltrationFiltration• HEPA (biological safety cabinets, ventilation)HEPA (biological safety cabinets, ventilation)• 0.2 micron (physically removes particulates )0.2 micron (physically removes particulates )


4. 4. Standard Operating ProceduresStandard Operating Procedures Physical Decontamination Physical Decontamination



4. 4. Standard Operating ProceduresStandard Operating Procedures Waste Disposal -AutoclavesWaste 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


Items that Items that CANCAN be autoclaved: be autoclaved:cultures and stocks of infectious materialcultures and stocks of infectious materialculture dishes and related devicesculture dishes and related devicesdiscarded live and attenuated vaccinesdiscarded live and attenuated vaccinescontaminated solid items (petri dishes, contaminated solid items (petri dishes, eppendorf tips, pipettes, gloves, paper towels)eppendorf tips, pipettes, gloves, paper towels)

Items that Items that CANNOTCANNOT be autoclaved: be autoclaved:chemical or radioactive wastechemical or radioactive wastecertain kinds of plasticscertain kinds of plastics




Items waiting for autoclaving are placed in Items waiting for autoclaving are placed in specially designated biohazard waste specially designated biohazard waste containers containing autoclave bagscontainers containing autoclave bags




PREPARATION OF WASTE:PREPARATION OF WASTE:

Use only approved autoclave bags.Use only approved autoclave bags.Do not overfill autoclave bagsDo not overfill autoclave bagsSeparate material for re-use from that which will be disposed and dry Separate material for re-use from that which will be disposed and dry from liquid material .from liquid material .If outside of bag is contaminated, double bag.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 occurAll flasks containing biological material should be capped with All flasks containing biological material should be capped with aluminum foil. aluminum foil.

Effectiveness of decontamination from a steam autoclave depends on appropriately loading items into the autoclave.






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


Treated waste (through autoclaving or other effective Treated waste (through autoclaving or other effective method of sterilization) is no longer considered method of sterilization) is no longer considered biohazardous and can be disposed in the regular waste biohazardous and can be disposed in the regular waste

streamstream

ALLALL biological waste must be treated prior to biological waste must be treated prior to disposal (disposal (including level 1 agentsincluding level 1 agents).).


4. 4. Standard Operating Procedures Standard Operating Procedures Waste DisposalWaste Disposal



4. 4. Standard Operating Procedures Standard Operating Procedures Waste Disposal –External Waste Disposal –External

ContractorContractorAny waste that cannot be treated (i.e. sharps, tissues) Any waste that cannot be treated (i.e. sharps, tissues) remains biohazardous waste and must be disposed off site remains biohazardous waste and must be disposed off site using a licensed waste disposal contractor.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


Continue on to:Continue on to:Large Scale WorkLarge Scale Work




4. 4. Standard Operating ProceduresStandard Operating Procedures Large Scale or High ConcentrationsLarge 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

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RYERSON UNIVERSITY 1 4. STANDARD OPERATING PROCEDURES.

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