Biosafety at Ryerson. 4. Standard Operating Procedures

Transcription

1 Biosafety at Ryerson 4. Standard Operating Procedures

2 Training Outline infection control containment of aerosols working in CL1 areas working in CL2 areas biological safety cabinets human blood and body fluids needle stick injury prevention signs and labels person protective equipment purchasing/transfer inventory control decontamination/waste disposal large scale work

3 Infection Control Good microbiological practice: is a basic code of practice to handle biological agents involves specific sterile techniques of product protection and contamination control is applied to all types of work involving microorganisms irrespective of containment level to reduce the risk of exposure and infection

4 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 surfaces & equipment exposure to biological agents infection

5 Infection Control: Hand Washing When to wash? before starting any manipulations before and after completing any task in a biosafety cabinet every time gloves are removed before contact with one s face or mouth before leaving the lab at the end of the day...and of course, when hands are obviously soiled

6 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 bar soaps.

7 Proper Hand Washing Techniques Wet hands with warm water Dispense soap into a cupped hand Spread around hands and between fingers as follows: Wash hands for at least 10 seconds Rinse thoroughly under warm water Dry hands thoroughly with paper towels

8 Containment of Aerosols

9 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.

10 Containment of Aerosols You cannot see the aerosol particles (the largest of them being 1/10th of a millimeter). Must become aware of what processes can generate aerosols. When in doubt, assume aerosols are generated. When conducting procedures that create considerable aerosols or when using agents classified as requiring Containment Level 2, work should be conducted in a Biological Safety Cabinet (BSC). When appropriate, other primary barriers such as splash shields, face protection or gowns should also be used.

11 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 include, but are not limited to: pouring liquids using centrifuges, shakers, blenders opening pressurized vessels inserting a hot loop into a culture pipetting, etc.

12 Containment of Aerosols Sterilization of inoculating 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 Containment of Aerosols: Pipetting mouth pipetting is prohibited all biohazardous materials should be pipetted in a BSC 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

14 Working in Containment Level 1 & 2 Areas

15 Working in CL 1 areas The following requirements are basic for any laboratory using biological agents: good microbiological laboratory practices intended to avoid the release of infectious agents a documented procedural (safety) manual must be available for all staff and its requirements followed the manual must be reviewed and updated regularly

16 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).

17 Working in CL 1 areas Laboratory must be kept neat and clean (good housekeeping). Storage of materials not pertinent, and that cannot be easily decontaminated, must be minimized. Paperwork and report writing should be kept separate from biological 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 (sharps container).

18 Working in CL 1 areas 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 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.

19 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 low 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. Keep lab coats in lab & street clothing outside lab. Suspected contaminated clothing must be decontaminated before laundering. Eye protection must be worn when necessary to protect the eyes. Gloves must be worn whenever there is a risk to the skin or risk from absorption through the skin on materials being handled. 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.

20 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.

21 Working in CL 1 areas Disinfection Disinfectants effective against the agents in use must be available at all times within the areas where the biological material is handled or stored. Effective concentrations and contact times must be used. All contaminated materials must be decontaminated before 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.

22 Working in CL 1 areas Accidents / Incidents All spills, accidents (needle sticks, 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: 1. Ryerson University s Internal Accident/ Exposure Investigation Report with Environmental Health and Safety (EHS) office within 24 hours of incident 2. Employer s Report of Injury/Disease (Form 7) WSIB Form 7 with Human Resources within 24 hours of incident Appropriate medical evaluation, surveillance, and treatment must be sought and provided as required. Actions taken to prevent future occurrences should also be documented.

23 Working in CL 2 areas In addition to the safety precautions outlined for Containment Level 1 laboratory additional operational practices for CL 2 labs are outlined below: Biological safety cabinets must be used for procedures that may produce aerosols and/or that involve high concentrations or Large Volumes ( 10 L) of biohazardous material/ This could be a single vessel with a volume of 10 litres or greater, or in some cases, multiple vessels with a total volume of 10 litres or greater. Determination of cutoff values for large scale volumes can be made in consultation with the PHAC or the CFIA/ Appropriate door sign must be posted outside each laboratory/

24 Biological Safety Cabinets

25 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, to product, and the environment.

26 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%.

27 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

28 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

29 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

30 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

31 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

32 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

33 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.

34 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.

35 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.

36 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

37 Human Blood and Body Fluids

38 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, etc.), tissue cultures, or organ cultures increases the risk pathogens of greatest concern are hepatitis B virus, hepatitis C virus and the Human Immunodeficiency Virus (HIV)

39 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

40 Human Blood and Body Fluids Universal precautions The minimum standard of practice to prevent exposure to bloodborne pathogens, including: 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.

41 Universal 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.

42 Needle Stick Injury Prevention

43 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.

44 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.

45 Needle Stick Injury Prevention To avoid needle stick injury: use needle-less techniques where possible use retractable sharps fill syringes carefully do not bend, or recap needles use approved designated sharps containers should be used for disposal that will not allow penetration or direct access to sharps

46 Signs and Labels

47 Signs and Labels Biohazardous or infectious materials fall under: Biohazardous Infectious Material of WHMIS Class 6.2 of Transport Canada s Transportation of Dangerous Goods Act (Infectious Substances)

48 Signs and Labels Entrances Biohazard signs must be posted on doors to rooms where biohazardous materials are used (Level 2 or greater)

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

50 Signs and Labels Inside lab (posted prominently) Internal Ryerson Biosafety Permit Current list of authorized users

51 Personal Protective Equipment

52 Personal Protective Equipment lab coat (buttoned) disposable gloves safety glasses footwear (closed toe) Responsibility of both the user and the supervisor to ensure that PPE is appropriately worn and kept in good condition

53 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.

54 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

55 Personal Protective Equipment Gloves nitrile and 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 Personal Protective Equipment Safety Glasses safety glasses or goggles to protect the eyes

57 Personal Protective Equipment Footwear closed toed shoes should always be worn sandals or open toed shoes are not allowed in a biohazard lab NO

58 Purchasing/Transfer

59 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.

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

61 Transfer of Human Cells/Tissue Outside the university prior BSO approval for lending or borrowing of biohazards from other institutions before arrival on campus receiving institution must have appropriate containment materials must be properly packaged and documentation persons preparing and/or transporting package must have valid TDG Class 6 Training certificate inventory must be adjusted

62 Inventory Control Permit Holder must have up to date inventory list of agents on permit, 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

63 Decontamination & Waste Disposal

64 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

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

66 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

67 Chemical Decontamination Choice depends on: type of material to be disinfected organic load chemical characteristics Most common are chlorine compounds and alcohols (broad range)

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

69 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 )

70 Sterilization by 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

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

72 Waste Disposal If you currently have solid waste: 1. Set up the supplied cardboard boxes with yellow plastic liner and place your bag inside (you can put multiple bags until the yellow liner/box is full) 2. Tie up liner, close and tape shut the box 3. If your waste contains cytotoxic drugs (i.e., chemo agents), put a cytotoxic label on the box 4. With a marker, write lab name and date on side of box (e.g., Smith lab/ Jan 12, 2017)

73 Solid Waste in Cardboard Boxes

74 Waste Disposal Supplies Colour Coding Yellow for regular biological waste Red for cytotoxic waste Any material that may have come into contact with a cytotoxic drug (such as chemotherapeutic agents) which inhibit or prevent the function of cells.

75 Waste Disposal Supplies Pails Use pails to collect waste that could puncture through plastic bags, such as serological pipettes, pipette tips, glass tubes, slides, etc. Use pails to collect liquids: do NOT pour liquids directly into pails, keep liquids in small bottles, then tighten their lids and place in pails

76 Waste Disposal Supplies Boxes 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

77 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 or contact EHS You can also ehs@ryerson.ca for more supplies if your department has run out

78 Waste Disposal Process When boxes are full, tie up liner and tape box closed When 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 and current date Example: Smith lab / Jan 12

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

80 Large Scale Work

81 Large Scale or High Concentrations Containment levels based on laboratory scale operations (in addition to pathogenicity of agents): 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

82 END