Presentation on theme: "Biosafety in Biomedical Laboratories June 2, 2009"— Presentation transcript:
1Biosafety in Biomedical Laboratories June 2, 2009 Marshall UniversitySummer Students/Recent HiresPlease complete the sign in sheet
2Major Safety Concerns Safe handling + containment of infectious agents Recombinant DNA (rDNA), D. Primerano June 2Management of Biohazard Waste (N. Douglas)Bloodborne Pathogen Exposure (N. Douglas)Chemical Safety (Nathan Douglas), June 3rdUse of Animals in Research(Dr. Billy Howard June 4)Radioisotope Use (Dr. Will McCumbee, June 8)Human Subject Protection (ORI, Mr. Bruce Day)
3Key Biosafety Personnel Chemical & Biological Safety OfficerMr. Nathan DouglasMarshall University Safety & HealthSorrell Maintenance Building, 215(304)(304) cellInstitutional Biosafety Committee, ChairDr. Donald Primerano, Room 336F BBSCEverything we cover will be posted to the IBC website
4Main Principle of Biosafety: Containment Purpose of Containment: to reduce or eliminate exposure of lab workers and others to hazardous biological agentsBacteria, viruses, fungi, parasitesRecombinant DNAPotentially dangerous cell lines
5General Principles of Biosafety Three elements of containment:Personal protective equipmentFacility designLab practice and techniquesExtent of containment depends onlevel of risk of exposurenature of agent
6Examples of Safety Equipment Biological Safety Cabinets (Laminar Flow Hood)Protect you and cells you are working with!!!Centrifuge Safety CanistersGloves, lab coats, gowns, face shield, goggles, shoe covers, respirators
7Biological Safety Cabinets (BSC) Class I and II BSCs provide effective containment system for safe handling of moderate/high risk organisms (BSL2 and BSL3 agents)Class II BSCs also protect the research material itself through high efficiency particulate filtration (HEPA) of the air flow down across the work surface (vertical laminar flow which forces outside air into the plenum)BBSC has shared facilities with Class II Type A2 BSC
9Facility Design and Construction (Secondary Barriers) Separation of lab work areas from areas of public accessAvailability of decontamination stations (autoclaves and hand-washing facilities)Separate ventilation systemsControlled access labs
10HOMEWORK ASSIGNMENTOn the way back to your lab today take note of the location ofShowersEye wash stationsFire extinguishersEmergency exit routes
11Biosafety Levels (BSLs) A biosafety level is the level of biocontainment precautions required to isolate dangerous biological agents in an enclosed facilityEach level represents a combination oflab practices,techniques,safety equipmentlab facilitiesdesigned to minimize release and exposure.
12Biosafety Level 1 (BSL 1)appropriate for work with organisms known to not cause disease in healthy adult humans.“Avirulent organisms”Minimal hazard to lab personnel and the environmentExamples:E coli, Bacillus spp,Exempt categories of rDNA work
13Biosafety Level 2 BSL 2: indigenous moderate risk agents Mild or treatable disease in humansNot spread by aerosol.Primary Hazard: Skin break, mucous membrane exposure or ingestion.Examples:Hepatitis A,B+C, HIV, some SalmonellaeHuman derived blood and blood productsCell culture work and some rDNA
14Biosafety Level 3 (BSL 3)work with indigenous or exotic agents with potential for respiratory transmission or lethal consequences.Examples:M. tuberculosis,B. anthracisSARS and West Nile virusesPrimary and Secondary barriers to protect personnel in contiguous areas
15Biosafety Level 4 (BSL4)BSL 4: lethal exotic agents especially where there is no vaccine or therapy.BSL 4 facility is separate facility or HVAC isolated zoneExamples: Ebola, Marburg and Lassa Fever virusesNO BSL3 or BSL4 work at MU.
16Which Biosafety Level Should You Be Using? (more homework) Ask your mentor first!!Read Sections I-III of Biosafety in Microbiological and Biomedical LaboratoriesGo to ABSA Website for a listing of infectious agents and corresponding BSLRead the NIH rDNA guidelines
17American Biological Safety Association (ABSA) If don’t which risk group your agent falls into:Call or Don Primerano ORConsult the ABSA website:
21Biosafety Level 1 Practices: “Standard Microbiological Practices” Access to lab is limited or restricted when experiments are in progressWorkers wash hands after handling viable organisms, after removing gloves and before leaving the labEating, drinking, smoking, handling contact lens, brushing teeth, storing food is absolutely forbidden.
22Biosafety Level 1Mouth pipetting is prohibited- YOU MUST USE MECHANICAL PIPETTING DEVICESSafe handling of sharps (disposal)Procedures minimize splashing and aerosolsWork surfaces are decontaminated with disinfectant on a daily basisAll cultures, stocks and other infectious materials are decontaminated by an approved method (usually autoclaving)Biohazard signs are posted at lab entranceInsect and rodent control program in place
23Biosafety Level 2Similar to BSL-1 and suitable for work with agents of moderate hazard.Lab personnel get specific training in handling pathogens.Access to the lab is limited when work is being conducted.Extreme precautions taken with contaminated sharps.Specific procedures for reducing aerosols or splashes.
24Biosafety Levels 3 and 4 Levels of personal protection increases Lab access is restrictedLab design becomes more criticalAnimal Biosafety Levels: special set of rules noted in the NIH rDNA guidelines
25General Biosafety Advice Discuss specific safety issues with your mentor and safety committee chairs.Lab heads should have written safety protocols that are lab/experiment specific.Read Biosafety in Microbiological and Biomedical Laboratories as needed.Contact Dr. Primerano at if you have questions or concerns.
26General Biosafety Advice Get a lab coat and safety equipment immediately!Don’t wear open shoes ever!Keep hair short or tied backGlove allergies: nitrile gloves may help
27Transformed or Cancer Cell Lines Any experiment involving transformed or cancer cell lines falls under Biosafety Level 2 (BL2) practices.Work with living cells should be done under a biological safety cabinet.Special precautions are taken to limit aerosols.Discarded media and cells are treated with 10% bleach before autoclaving (sterilization).Decontaminate spills with bleach and wipe down hood after working with cells.Protective clothing (lab coat and gloves) should be worn when working with the cells.Restrict access to the lab when working with cell lines.
29Recombinant DNA (rDNA) as defined by the NIH Guidelines rDNA molecules are either:DNA molecules constructed outside cells by joining nature or synthetic DNA segments to DNA molecules that can replicate in a living cell, ORMolecules that result from the replication of those described above
30rDNA Regulations Follow BSL1-4 guidelines. Each lab must have approved protocols for use of rDNA.Application forms and guidance on the IBC websiteBSL1 is appropriate for most rDNA work at Marshall (but not all).Consult mentor or rDNA guidelines for all casesEach investigator (your mentors) must be familiar with the guidelines.
32Bloodborne Pathogen Standard Exposure Control Plan Organisms in human blood that cause disease. Includes bacteria and viruses like HbV and HIVOSHA requires that the university have an Exposure Control Plan (on IBC web page)Also requires employers to provide Hepatitis B vaccination at no cost to employee
33Bloodborne Pathogen Standard Universal PrecautionsTreat all human blood and other body fluids as if they are contaminated with an infectious agent.Required by the OSHA BBPSUse Personal Protective Equipment (gloves, face mask, lab coat) when around biohazard materialsDo not take lab coat home, BBSC will launder
34Bloodborne Pathogen Standard No eating, drinking, smoking, applying cosmetics or contact lenses in the labsNo mouth pipetting; use pipetting deviceProper disposal of blood and other biohazard materialsBleach (1:10 solution of bleach and water)Autoclave on-siteOff-site shipment for treatment and disposal
35Bloodborne Pathogen Standard If you are working with human blood, you must read and understand the Exposure Control Plan AND meet with Dr. Primerano!Immediately report exposures to Dr. Donald Primerano (w) or (h).Sets in motion the required follow up steps
36Infectious Medical Waste Rule WV Department of Health & Human Resources
37Infectious Medical Waste Rule 1. Definition of Infectious Medical Waste2. Types of Infectious Waste3. WV DHHR License4. Infectious Waste Disposal Plan5. Bloodborne Pathogen Standard Review
38Infectious Medical Waste Rule WV Legislative Rule 64 CSR 56Section Infectious Medical Waste is medical waste which is capable of producing an infectious disease.
39Infectious Medical Waste Rule Medical waste shall be considered capable of producing an infectious disease ifit has been, or is likely to have been, contaminated by an organism likely to be pathogenic to healthy humans,if such organism is NOT routinely and freely available in the community, ANDsuch organism has a significant probability of being present in sufficient quantities and with sufficient virulence to transmit disease.
40Infectious Medical Waste BBSC Plan: Sterilize all bacterial, fungal, viral and parasitic organisms and cultured cells used in research.Includes deliberately infected animalsInfected animals go directly to StericycleIf you have any questions please consult with Don Primerano ( ) or Connie Berk ( ).
41Types of Infectious Waste Cultures of microorganisms & biologicalsHuman blood and blood productsAll contaminated sharpsPathological wastesContaminated animal carcasses, body parts, bedding and related wastesCleanup materials from an infectious medical waste spillWaste contaminated by, or mixed with, infectious medical waste
42Cultures and Stocks of Microorganisms and Biologicals
43Human Blood & Blood Products All human blood (wet or dried)Products from human blood (plasma)Does not include animal blood
44Sharps Any article that can puncture or cut If sharps have been used in patient care or treatment OR used to handle or deliver infectious agents, must be placed in a “Biohazard” labeled sharps box or broken glass containerSharps boxes & containers will be autoclavedExamples: pipette tips, glass Pasteur pipettes, needles, syringes, scalpel blades, razors, forceps
45Pathological WasteIncludes: tissues, organs, body parts, and containers of body fluidsAll pathological waste should be packaged by the investigator and picked up by StericycleStericycle provides shipping containers and liners.Containers and liners are located in BBSC Room Contact Julia Schreiber or Connie Berk if you need packing materials.
46Animal WasteContaminated animal carcasses, body parts, animal bedding known to have been exposed to infectious agents during researchThese items should be taken to the animal facility where Julia or Connie will assist you in preparing for pickup by Stericycle.
47Infectious Medical Waste Management Permit BBSC permit from the WV DHHR is renewed annuallyAllows us to generate infectious waste, sterilize on site via autoclave, and dispose waste in regular trashAllows us to hire a licensed company to remove infectious waste that cannot be autoclaved (e.g. contaminated animal carcasses)Our company is Stericycle
48Infectious Medical Waste Management Plan OBJECTIVES:Provide a safe environment for faculty, staff, students and visitorsProperly manage infectious waste in accordance with WV Legislative Rule 64 CSR 56, Infectious Medical Waste
49Infectious Medical Waste Management Plan Handling of infectious wastePackagingTransportationSterilizationStorage and disposalContingency planSpill responseTraining
50Infectious Waste Packaging Orange biohazard bags must be used for infectious waste, clear and red bags are not acceptableWaste collection containers and bags must be double-baggedDo not fill them more than 2/3 fullLoosely gather the top of the bag and place a strip of autoclave tape around it, do not tie the bags closed. Steam must be able to enter the bag.Bags, flasks, and other containers of infectious material must be labeled with primary investigator’s name, room number, phone number, and the contents.
51Special Packaging Rules Labs must have separate containers for contaminated and uncontaminated glasswareBroken glass boxes are acceptable, and must include a biohazard sticker when used for contaminated itemsContaminated pipette tips can easily puncture plastic bags. They must be collected in a sharps box, or in an orange bag, the bag put into a cardboard box, then placed into a biohazard waste collection containerContaminated Pasteur pipettes must go into a double-bagged, biohazard-labeled contaminated glassware boxUncontaminated glass must go into a box labeled as “Broken Glass”
52Transporting Waste To Autoclave Room 119A All waste must be transported on/in carts with secondary containment trays.Infectious waste may not be sterilized in the satellite autoclaves on the floors.Bags of solid infectious waste must be placed in the large gray bin in Room 119A, accessible 24/7. Lab heads will provide labels and keys to gray bin. Relock the bin, which must be kept locked at all times.Liquid infectious waste must be placed on the metal cart.Waste will be autoclaved, weighed (if solid) and placed into the regular trash. We weigh waste as required by our state permit.
53Sterilization of Infectious Waste Infectious waste must be autoclaved for 90 minutes.Autoclaved waste is placed in black plastic bags, weighed, and taken to the dumpster.Sterilized reusable items (glassware, etc.) are placed at the pickup point in the media washing room, BBSC 121.
54Contingency PlanIn the event that waste cannot be autoclaved at the BBSC, Stericycle will be employed to treat and dispose of the waste.Waste items sent for treatment and disposal via Stericycle must be packaged and labeled by someone properly trained in DOT Hazardous Materials.
55Spill ResponseNotify Dr. Primerano or Connie Berk as soon as possible.Don: (w) or (h)Connie: (w) orIf you cannot reach either of them, contact Nathan Douglas (304) ,Spill kits are located in each satellite autoclave room on every floor, as well as in room 119 and room 121.
56Spill ResponseAttempt to prevent liquids from running free by laying paper towel or other absorbent material on top.Do not use spill cleanup kit unless you are comfortable doing so, allow someone trained to clean spill. Make sure you reach someone right away in either case.Complete a Biohazard Spill Occurrence form, available on IBC web page:
57Don’t Put Noninfectious Items In Orange Bags Bubble wrapPaper towels, either from drying your hands or from wiping down benches with a disinfectant Scalpel blade wrappers Needle wrappers Benchkote or any other bench protector Gloves, unless contaminatedPipettesFood or drink containers (not allowed in labs)Packaging materials such as cardboard, Styrofoam “peanuts”, etc.Paper (copy paper, etc.)Pop cansAll of the above items can be recycled in the blue bins in the middle hallway.
58ReviewWe care about biosafety to prevent the spread of bloodborne pathogensOSHA Standard covers HbV & HIVEveryone exposed should have completed the 3-shot Hepatitis B vaccine seriesGloves are used to protect us as we workGloves must be removed before exiting laboratories, and hands should be washed.If items must be transported, use a cart.If you must carry something, use only 1 gloved hand, use the other to open doors.