Presentation on theme: "Biological Laboratory Safety University of Colorado Colorado Springs"— Presentation transcript:
1 Biological Laboratory Safety University of Colorado Colorado Springs Environmental Health and Safety
2 Biological Laboratory Safety Training IntroductionThe purpose of this training module is to familiarize the Principal Investigator and lab personnel with good microbiological practices which include recognizing risk groups for biological materials, appropriate containment levels, and personal protective clothing and equipment.
3 Biological Laboratory Safety Training Type of ResearchDepending upon the type of research being conducted, a particular protocol may be subject to review by one or more of the following groups:Research Involving Human Subject (IRB) Research Involving Animals (IACUC) Research Involving Biohazards (IBC)
4 Research Involving Human Subject (IRB) Biological Laboratory Safety TrainingResearch Involving Human Subject (IRB)The participation of humans in research projects raises fundamental ethical and civil rights questions. All such research, funded and unfunded projects sponsored and not sponsored which is carried out by students, faculty, or other University employees, on or off campus, is covered by the Policies and Procedures of the UCCS Institutional Review Board (IRB) for the Protection of Human Subjects.
5 Research Involving Animals (IACUC) Biological Laboratory Safety TrainingResearch Involving Animals (IACUC)Several documents and organizations provide policies, standards and regulations on the care and use of animals national wide. The following is a summary of those national guidelines that are pertinent to researchers using animals at the University of Colorado at Colorado Springs.
6 Research Involving Biohazards (IBC) Biological Laboratory Safety TrainingResearch Involving Biohazards (IBC)The University of Colorado Colorado Springs has established an Institutional Biosafety Committee (IBC) whose responsibilities need not be restricted to recombinant DNA and shall meet the requirements as outlined in Section IV-B-2 of the National Institutes of Health (NIH) Guidelines for Research Involving Recombinant DNA Molecules.This is the focus of this training
7 Biological Laboratory Safety Training What is Biosafety?The measures employed to avoid infecting oneself, others or the environment when handling biohazard materialsPracticing good microbiological techniques is the single most effective means of preventing exposures
8 Why Biosafety Practices? Biological Laboratory Safety TrainingWhy Biosafety Practices?Protection for:- workers- “products”- co-workers- lab support personnel- lab or field animals- environment
9 Biological Laboratory Safety Training BiohazardAn agent or material of biological origin that has the capacity to produce deleterious effects on humans, i.e. microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from higher plants and animals.Examples:Viruses, bacteria, fungi, and parasitesBlood and body fluids, as well as tissues from humans and animalsTransformed cell lines or human cells
10 Biological Laboratory Safety Training PathogenA disease causing agent.Most are infectious microbes, such as bacteria or viruses, which are capable of causing disease. Other parasites, such as fungi and protozoans, are also considered pathogens. Because not all microbes are harmful, pathogens refer specifically to those that can cause disease or other harm.Examples:Viruses, bacteria, fungi, and parasites
11 Biological Laboratory Safety Training Infectious AgentAn agent of biological origin that is transmissible through contact, ingestion, inhalation, or animal and insect bites that may cause disease. If they are pathogenic then they will cause an infectious disease.
12 Biological Laboratory Safety Training Zoonotic DiseaseAn infectious disease that can be transmitted from animals to humans.A number of infectious diseases, including viruses, bacteria, and parasites, can be transmitted from animals to people through a variety of infection routes, including animal bites, vectors (i.e., insects), and animal-to-human contact (i.e., inhalation of respiratory droplets or skin-to-skin contact). Some examples of common zoonotic diseases include lyme disease, rabies, ringworm, and plague.
13 Classification of Agents Biological Laboratory Safety TrainingClassification of AgentsThe NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules established a classification and assigned human etiological agents into four risk groups on the basis of hazard and requirements for each biosafety level of containment.The CDC guideline, Biosafety in Microbiological and Biomedical Laboratories (BMBL) uses the World Health Organization (WHO) recommendations for an agent risk group classification for laboratory use that describes four general risk groups requirements for each biosafety level of containment.
14 Risk Group Classifications Biological Laboratory Safety TrainingRisk Group ClassificationsA list of Risk Group Agents can be found in the NIH Guidelines:A list of Agent Summary Statements can be found in the BMBL:Risk Group ClassificationNIH Guidelines for Research Involving Recombinant DNA Molecules, 2011WHO Laboratory Biosafety Manual,3rd EditionRisk Group 1Agents that are not associated withdisease in healthy adult humans.(No or low individual and community risk) Amicroorganism that is unlikely to cause human oranimal disease.Risk Group 2Agents that are associated with humandisease which is rarely serious and forwhich preventive or therapeuticinterventions are often available.(Moderate individual risk; low Community risk) Apathogen that can cause human or animal diseasebut is unlikely to be a serious hazard to laboratoryworkers, the community, livestock or the environment.Laboratory exposures may cause serious infection, butEffective treatment and preventive measures areavailable and the risk of spread of infection is limited.
15 Which Guidelines Should I Follow? Biological Laboratory Safety TrainingWhich Guidelines Should I Follow?For all rsNA activities (includes transgenic animalsand plants):The NIH Guidelines for Research InvolvingRecombinant or Synthetic Nucleic Acid MoleculesFor all non-rsNA activities:The CDC guideline, Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th Edition
16 Risk Group Classifications Biological Laboratory Safety TrainingRisk Group ClassificationsCurrently, UCCS prohibits the possessionor use of Risk Group 3 or 4 AgentsRisk Group ClassificationNIH Guidelines for Research Involving Recombinant DNA Molecules, 2011WHO Laboratory Biosafety Manual,3rd EditionRisk Group 3Agents that are associated with serious orlethal human disease for which preventive ortherapeutic interventions may be available(high individual risk but low community risk).(High individual risk; low community risk) Apathogen that usually causes serious human oranimal disease but does not ordinarily spreadfrom one infected individual to another. EffectiveTreatment and preventive measures are available.Risk Group 4Agents that are likely to cause serious orLethal human disease for which preventive ortherapeutic interventions are not usuallyavailable (high individual risk and highCommunity risk).(High individual and community risk) A pathogenthat usually causes serious human or animaldisease and that can be readily transmitted fromone individual to another, directly or indirectly.Effective treatment and preventive measures arenot usually available.
17 Biological Laboratory Safety Training Risk Group AgentsExamples of Risk Group 1 AgentsBacillus subtilisEscherichia coli K-12Pichia pastorisExamples of Risk Group 2 Agents:Acinetobacter baumanniiSalmonella typhimuriumStaphylococcus aureusCryptosporidium including C. parvum amphotropic and xenotropic strains of murine leukemia virusHepatitis A, B, C, D, and E virusesEpstein Barr virusHuman cells, blood, blood products, bodily fluids, tissue
18 Biological Laboratory Safety Training Risk Group AgentsExamples of Risk Group 3 Agents:Burkholderia (Pseudomonas) mallei, B. pseudomalleiYersinia pestisHistoplasma capsulatum, H. capsulatum var.. duboisiiFlaviviruses (Togaviruses) - Group B ArbovirusesPrionsRetrovirusesHuman immunodeficiency virus (HIV) types 1 and 2Human T cell lymphotropic virus (HTLV) types 1 and 2Simian immunodeficiency virus (SIV)Examples of Risk Group 4 Agents:Ebola virusSmallpoxHerpesvirus simiae (Herpes B or Monkey B virus)Hemorrhagic fever agents and viruses as yet undefined
19 Routes of Transmission Biological Laboratory Safety TrainingRoutes of TransmissionMost common routes of transmission in laboratory which can lead to laboratory acquired infections:direct skin, eye or mucosal membrane exposure to an agent;parenteral inoculation by a syringe needle or other contaminated sharp, or by bites from infected animals and arthropod vectors;ingestion of liquid suspension of an infectious agent, or by contaminated hand to mouth exposure;inhalation of infectious aerosols; and vectors, such as mosquitoes, ticks, etc.
20 Biological Laboratory Safety Training Risk AssessmentOnce the investigator has decided on the agent, recombinant or synthetic molecule, then he or she must conduct an assessment of risk.1) Identify agent hazards and perform initial assessment of risk which includes:agent in use/use of recombinant or synthetic nucleic acid molecule materialsvirulence/pathogenicity/infectious doseenvironmental stability e.g. temperature, sterility, UV sensitivitymode of transmission e.g. contact, ingestion, inhalation, animal bitequantity/concentration/volume used (additional requirements >10L, notify Biosafety Officer)vaccine/treatment availabilityallergenicity
21 Biological Laboratory Safety Training Risk Assessment2) Identify laboratory procedure hazards which may include:use and disposal of sharpsaerosol generating procedures e.g. sonication, vortexing, homogenization, flow cytometry, pipettingvolume of culture used (additional requirements >10L, notify Biosafety Officer)transport of agentsuse of chemicals and radionuclidesuse of animalswork on open bench topdisposal of infectious waste
22 Biological Laboratory Safety Training Risk AssessmentMake a determination of appropriate biosafety level of containment and select additional precautions indicated by risk assessment.Evaluate proficiencies of staff regarding safe practices and integrity of safe equipment.Review risk assessment with Biosafety Officer, subject matter expert, and IBC.
23 Biological Laboratory Safety Training ContainmentThe objective of a good lab biosafety program is the containment of potentially harmful biological agents and materials.Describes safe methods, facilities and equipment for managing infectious materials in the lab where they are being handled or maintained.Reduces or eliminates exposure of lab workers, other persons, laboratory animals, and the outside environment to potentially biohazardous agents and materials.
24 Biosafety Levels of Containment Biological Laboratory Safety TrainingBiosafety Levels of ContainmentThere are four Biosafety Levels:BSL1 - agents not known to cause diseaseBSL2 - agents associated with human disease which are easily treatable or prevented by vaccinationBSL3 - indigenous/exotic agents associated with human disease and with potential for aerosol transmissionBSL4 - dangerous/exotic agents of life threatening nature
25 Biosafety Levels of Containment Biological Laboratory Safety TrainingBiosafety Levels of ContainmentAs the level Moves UPthe risk of the organism to humans, animals, plant and/or the environment increasesthe procedural and facility requirements increasesthe level of containment required increasesthe degree of protection for personnel, the environment and the community increasesBSL-4BSL-3BSL-2BSL-1
26 BMBL Summary of Biosafety Levels Biological Laboratory Safety TrainingBMBL Summary of Biosafety Levels
27 General Lab Requirements Biological Laboratory Safety TrainingGeneral Lab RequirementsAn Approved Institutional Biosafety Committee (IBC) Application is Required for All Biological ResearchKnowledgeable Principal Investigator and Lab Personnelaware of potential hazardsproficient in practices & techniquesAdministrative Controlsappropriate signage (emergency and biohazard notifications signs, biohazard, biohazardous waste, non-infectious stickers, etc.)training
28 General Lab Requirements Biological Laboratory Safety TrainingGeneral Lab RequirementsBiosafety Levels of Containment (BSLs)Laboratory Practice & Techniquestandard practicesspecial practicesSafety Equipment (Primary Barriers)Facility or Laboratory Design and Construction (Secondary Barriers)
29 General Lab Requirements Biological Laboratory Safety TrainingGeneral Lab RequirementsBiosafety Cabinets (BSCs) – Required for BSL-2 whenever aerosol generating procedures are performedPersonal Protective Clothinggloveslab coatseye and face protectionPipetting DevicesSafety Centrifuge Cups and Rotors
30 Biological Laboratory Safety Training Primary BarriersSafety Equipment (e.g. Biological Safety Cabinet) and Personal Protective Equipment (PPE) are used to protect lab personnel working directly with infectious materials (e.g. gloves, safety glasses, lab coat).Secondary BarriersFacility or Laboratory Design and Construction are used to protect from exposure to infectious materials.
31 Biosafety Level 1 (BSL-1) Introduction Biological Laboratory Safety TrainingBiosafety Level 1 (BSL-1) IntroductionSuitable for work involving well characterized agents not known to cause disease in healthy adult humans and of minimal potential hazard to laboratory personnel and the environment.Examples:Bacillus subtilisInfectious canine hepatitis virusE. coli (non-infectious common lab strains, i.e. DH5alpha, K-12)
32 Biosafety Level 1 (BSL-1) Standard Work Practices Biological Laboratory Safety TrainingBiosafety Level 1 (BSL-1) Standard Work PracticesLimited or restricted access when experiments are in progressDecontaminate work surfaces daily and after a spillUse mechanical pipetting devicesNo eating or drinking in work areaWash hands frequentlyMinimize splashes and aerosolsWritten procedures for safe handling of sharps (i.e. needles, scalpels, razor blades, broken glass, pipettes)
33 Biosafety Level 1 (BSL-1) Standard Work Practices Biological Laboratory Safety TrainingBiosafety Level 1 (BSL-1) Standard Work PracticesPosted Biohazard Notification sign on lab entry door(s).PI must ensure lab personnel receive appropriate training regarding their duties, precautions to prevent exposures, and exposure evaluation proceduresFollow the EHS Biowaste Policy (please take a moment and review this policy)Contact Facilities Services for insect or rodent issues – (719)
35 Biosafety Level 1 (BSL-1) Laboratory Design (Secondary Barrier) Biological Laboratory Safety TrainingBiosafety Level 1 (BSL-1) Laboratory Design (Secondary Barrier)Requirements:Laboratories have doorsSink for hand washingWork surfaces & floors easily cleaned and decontaminatedBench tops are impervious to waterSturdy furnitureWindows fitted with fly screensNo special ventilationNormal building construction
36 Biosafety Level 2 (BSL-2) Introduction Biological Laboratory Safety TrainingBiosafety Level 2 (BSL-2) IntroductionSuitable for work involving well characterized agents that are associated with human disease that is rarely serious and for which preventative or therapeutic interventions are often available to laboratory personnel and the environment.Examples:Schistosoma - including S. haematobium, S. intercalatum, S. japonicum, S. mansoni, S. mekongiSalmonella – all speciesHuman cells or cell lines, bodily fluids, tissue
37 Biosafety Level 2 (BSL-2) Standard Work Practices Biological Laboratory Safety TrainingBiosafety Level 2 (BSL-2) Standard Work PracticesSame as BSL-1 PracticesSpecial Work PracticesRestricted lab access- limited or no access if immunocompromised- medical surveillance of lab personnel as appropriateLab personnel- aware of potential hazards- proficient in practices/techniques
38 Biosafety Level 2 (BSL-2) Special Work Practices Biological Laboratory Safety TrainingBiosafety Level 2 (BSL-2) Special Work PracticesBiosafety manual specific to lab and biological research- Biohazard Control Plan (part of IBC application)- Post Exposure Plan (may be required as part of IBC application depending on biological agents and materials used)Report accidents immediately- seek medical attention- notify supervisor or Principal Investigator- notify EH&S ( )- complete Risk Management Employee Injury Report https://www.cu.edu/risk/services/workers-compensationor Needle Stick or Bodily Fluid Exposure Formhttps://www.cu.edu/risk/services/workers-compensation
39 Biosafety Level 2 (BSL-2) Safety Equipment (Primary Barrier) Biological Laboratory Safety TrainingBiosafety Level 2 (BSL-2) Safety Equipment (Primary Barrier)Same as BSL-1 Safety Equipment PLUSBiosafety Cabinet- cabinet must be certified annually- UV light is strongly discouraged- properly disinfect before and after each useEye, face, and respiratory protection used in rooms containing infected animals as determined by risk assessment
40 Biosafety Level 2 (BSL-2) Laboratory Design (Secondary Barrier) Biological Laboratory Safety TrainingBiosafety Level 2 (BSL-2) Laboratory Design (Secondary Barrier)Self-closing doors with locksSink for hand washing located in labSpaces between benches, cabinets, and equipment must be accessible for cleaningBench tops, work surfaces, chairs must be non-porous and can be decontaminatedAll operable windows must remain closed or fitted with screens when work is performedEmergency eye/face wash unit must be availableVacuum lines with liquid disinfectant traps or in-line filterAvailable autoclave for disinfectionTrash can with foot operated lid
41 What Do I Do if I’m Exposed to a Biohazard that IS NOT a Pathogen? Biological Laboratory Safety TrainingWhat Do I Do if I’m Exposed to a Biohazard that IS NOT a Pathogen?Graduate Student, Teachers Assistant (TA) or Staff Employees PAID by the University of Colorado:If you are exposed to a biohazardous agent in the course of your normal duties, you qualify to be treated under workman’s compensation and may elect to go to one of the two Work Comp providers in Colorado Springs:CONCENTRA MEDICAL CENTER CENTURA CENTERS FOR OCCUPATIONAL MEDICINE (CCOM)5320 Mark Dabling Boulevard Union Medical Campus (NW Fillmore)Building 7, Suite Medical Center Point, #100(719) (719)OR - Penrose or Memorial Hospital Emergency Departments when the Work Comp providers are closed or unable to see you at the time of your exposure
42 What Do I Do if I’m Exposed to a Biohazard that IS NOT a Pathogen? Biological Laboratory Safety TrainingWhat Do I Do if I’m Exposed to a Biohazard that IS NOT a Pathogen?Undergraduate and Graduate Student, Teachers Assistant (TA) or Staff Employee NOT PAID by the University of Colorado:If you are exposed to a biohazardous agent, you may elect to go to anyone who is covered by your health insurance.- if you are a registered student at UCCS, you may go to the Student Health Center to do post-exposure testing
43 What Do I Do if I’m Exposed to a Biohazard that IS a Pathogen? Biological Laboratory Safety TrainingWhat Do I Do if I’m Exposed to a Biohazard that IS a Pathogen?Graduate Student, Teachers Assistant (TA) or Staff Employee PAID by the University of Colorado:If you are exposed to a biohazardous agent that is a pathogen or may contain a pathogen in the course of your normal duties, you qualify to be treated under workman’s compensation and should go directly to Penrose Hospital or Memorial Hospital Emergency Department- bring Post Exposure Plan and any information about agent(s) that you were exposed to with you
44 What Do I Do if I’m Exposed to a Biohazard that IS a Pathogen? Biological Laboratory Safety TrainingWhat Do I Do if I’m Exposed to a Biohazard that IS a Pathogen?Undergraduate and Graduate Student, Teachers Assistant (TA) or Staff Employee NOT PAID by the University of Colorado:If you are exposed to a biohazardous agent that is a pathogen or may contain a pathogen in the course of your normal duties, you may elect to go to anyone who is covered by your health insurance.- if you are a registered student at UCCS, you may go to the Student Health Center to do post-exposure testing or Penrose or Memorial Hospital Emergency Departments when the Health Center is closed or unable to see you at the time of your exposure- bring Post Exposure Plan and any information about agent(s) that you were exposed to with you
45 Safe Use of Centrifuges Biological Laboratory Safety TrainingSafe Use of CentrifugesBefore use, check to see if:balanced – may need a “placeholder tube”overfilledcaps or stoppers properly in placerun conditions (rpm – revolutions per minute vs. rcf - relative centrifugal force)Use sealable buckets (safety cups) or sealed rotorsAfter run, check to see if:centrifuge completely stoppedspills or leaks (clean immediately with appropriate disinfectant)allow aerosols to settle (30 minutes) or open in a Biosafety Cabinet
46 Biological Laboratory Safety Training Ultra Violet LightUV lights are strongly discouraged:Should never replace chemical surface disinfectionDust and debris collected on UV bulb will inhibit effectivenessThey provide a false sense of security - not giving off UV light (just prior to the end life of the bulb)Only disinfects the surface – could contact a layer of media protein covering the pathogenCasts a shadow, leaving areas that remain unexposed to the UV wavelengthsCould cause burns / damage eyes –turn off when working in the room
47 Biological Laboratory Safety Training Needles and SyringesAvoid use whenever possibleUse a biosafety cabinet for all operations with infectious agents or materials that may cause diseaseFill syringes carefullyShield needles when withdrawing from stoppersDo not bend, shear or recap needlesDispose of all used needles and syringes in a sharps container
48 Biological Laboratory Safety Training PipettesMouth pipetting is prohibitedAll biohazardous materials should be pipetted in a biosafety cabinetNever ‘blowout’ (force) fluids out, use ‘to deliver’ pipettes (pipette held vertically with tip against side of receiving vessel to drain completely)To avoid splashes, allow discharge to run down the receiving container wallNever mix material by suction and expulsionReusable pipettes should be placed horizontally in a disinfectant filled pan and autoclaved before reuse
49 Blenders, Grinders, Sonicators and Lyophilizers Biological Laboratory Safety TrainingBlenders, Grinders, Sonicators and LyophilizersOperate in a biosafety cabinet (except lyophilizer) whenever possible. Allow aerosols to settle for 5 minutes before openingSafety blenderdo not use glass blender jarsdecontaminate immediately after use with appropriate disinfectantLyophilizersuse glassware designed for vacuum workensure there is no damage before usingdisinfected all surfaces immediate after use with appropriate disinfectantuse vapor traps whenever possible
50 Biological Laboratory Safety Training Inoculation LoopSterilization in an open flame may create aerosols which may contain viable micro organismsUse a shielded electric incineratorShorter handles minimize vibrationsDisposable plastic loops are good alternatives
51 Biological Laboratory Safety Training CryostatsWear gloves during preparation of frozen sections and heavy gloves when accessing the cryostatDecontaminate frequently or immediately after use with biohazardous agent or material. Use an appropriate disinfectant that is specifically effective for the agent.
52 Biological Laboratory Safety Training Export ControlsDoes your research involve…Export or transport of material outside the US?Working with foreign nationals, institutions, or students?If you answered “yes” to either of these questions, contact Cindy Norton at or
53 Biological Laboratory Safety Training QUESTIONS?Contact InformationSandy Berry-LoweIBC ChairDepartment of Biology(719)Cynthia NortonInstitutional Biosafety OfficerEnvironmental Health and Safety(719)
54 THANK YOU You are now ready to take the required Biosafety Quiz Biological Laboratory Safety TrainingTHANK YOUYou are now ready to takethe required Biosafety Quiz