2 What is Biosafety?Safety from exposure to Infectious AgentsSmallpox
3 Did the Plague Kill Illinois Scientist? What is Biosafety?Sunday, Sep. 20, 2009Did the Plague Kill Illinois Scientist?By AP(AP / CHICAGO) — The University of Chicago Medical Center says the infection that killed a scientist may be connected to bacteria he researched that causes the plague.The university said Saturday that its researcher studied the genetics of harmful bacteria including Yersinia pestis, which causes the illness. He died Sept. 13. His name and age haven't been releasedThe medical center says the bacteria he worked with was a weakened strain that isn't known to cause illness in healthy adults. The strain was approved by the Centers for Disease Control and Prevention for laboratory studies.An autopsy found no obvious cause of death but did find the presence of the bacteria. More tests are planned. No other illnesses have been reported.
4 Biosafety in Various Disciplines Biosafety is related to several fieldsECOLOGY: referring to imported life forms not indigenous to the region (Reggie the alligator)AGRICULTURE: reducing the risk of alien viral or transgenic genes, or prions such as BSE/"MadCow“; reducing the risk of food bacterial contaminationMEDICINE: referring to organs or tissues from biological origin, or genetic therapy products, virus; levels of lab containment protocols BSL-1, 2, 3, 4 in rising order of dangerCHEMISTRY: i.e., nitrates in water, PCB levels affecting fertilityEXOBIOLOGY: i.e., NASA's policy for containing alien microbes that may exist on space samples - sometimes called "biosafety level 5"
5 Biosafety in Academic Research Research Universities:Promoting safe laboratory practices, and procedures; proper use of containment equipment and facilities; provides advice on laboratory design and risk assesment of experiments involving infectious agents, rDNA in-vitro and in-vivo.Bottom Line: Risk & Containment
6 Biohazard Symbol Charles Baldwin at National Cancer Institute at NIH. Symbol to be “memorable but meaningless” so it could be learned.Blaze orange – most visible under harsh conditions
12 Biohazardous Materials Human and Primate Cells, Tissues, and Body FluidsBrain Tissue from Demented PatientsViral VectorsReplication deficient viruses
13 Biosafety Concepts Biosafety In Microbiological and Biomedical Laboratories“BMBL” (acronym)CDC/NIH PublicationSafety “Guidelines”Regulations of Institution receivesNIH fundingCode of Practice and “Gold” Standard in Industry anlGold StandardClinical & Research Lab.Lab. Animal FacilitiesThe 4th edition of the BMBL (1999) contained the first government writings about security concepts for Select Agents. By inclusion, these ideas can be used by all laboratories that store and use pathogens, particularly those that might be perceived as or used for terrorist purposes.HHS Publication No. (CDC)
14 The New BMBLEarly print edition….Emphasis on “Risk& Containment”
15 Biosafety Concepts The BMBL The BMBL continuesto be published by theCDC and the NIH5th edition is now at the printers
16 Are the NIH Guidelines Optional? “Guidelines” does not mean “optional”They are a term and condition of NIH funding for recombinant DNA research.Now although they are called ‘Guidelines’ we don’t want to give you the impression that they are optional. You can’t pick and choose which sections to follow based on what sounds like a good idea or seems to much trouble to deal with.The Guidelines are a term and condition of NIH funding for recombinant DNA research and failure to comply with them can affect the funding for recombinant DNA work at an institution.From Kathryn Harris, NIH, OBA
17 Biosafety Concepts from the BMBL Principles of BiosafetyPractice and ProceduresStandard PracticesSpecial Practices & ConsiderationsSafety EquipmentFacility Design and ConstructionIncreasing levels of protection
18 Principles of Biosafety Biosafety Levels 1-4 (BSL)Increasing levels of employee and environmental protectionGuidelines for working safely in research & medical laboratory facilitiesAnimal Biosafety Levels (ABSL)Laboratory animal facilitiesAnimal models that support researchGuidelines for working safely in animal research facilities
19 Biosafety Concepts The BMBL (1) Standard Microbiological PracticesMost important concept / Strict adherenceAware of potential hazardTrained & proficient in techniquesSupervisors responsible for:Appropriate Laboratory facilitiesPersonnel & TrainingSpecial practices & precautionsOccupational Health Programs
20 Biosafety Issues The BMBL (2) Safety EquipmentPrimary Containment BarrierMinimize exposure to hazardPrevent contact / Contain aerosolsEngineering controls/ equipmentPersonal Protective Equipment (PPE)Gloves, gowns, Respirator, Face shield, BootiesBiological Safety CabinetsCovered or ventilated animal cage systems
21 Biosafety Concepts The BMBL (3) Facility Design and ConstructionSecondary Barrier/ Engineering controlsContributes to worker protectionProtects outside the laboratoryEnvironment & NeighborhoodEx. Building & Lab design, Ventilation, Autoclaves, Cage wash facilities, etc.
24 DiscussionWhat are some of the negatives and positives of this open lab concept?
25 Biosafety Level-1 Concepts of Biosafety Biosafety Level-1 (BSL-1 or ABSL-1)Well characterized agentsAgents not known to cause disease (in healthy human adults; now healthy immunocompetent adults)Prophylactic treatment availableOpen bench proceduresAnimals in open cage system or open environment (outdoors)Good laboratory practices
27 BSL-1 Practices Bench-top work allowed Daily Decontamination Manual pipettingRequired HandwashingRed bag wasteBio cabinet not required (unless creating aerosols)2˚ containment
28 Risk Group 2 Agents Human or Primate Cells Herpes Simplex Virus Replication Incompetent Attenuated Human Immunodeficiency VirusPatient specimens
29 BSL-2 Practices Concepts of Biosafety Practices & ProceduresAgents associated w/ human diseaseTreatment for disease availableAgent poses moderate hazard to personnel and environmentDirect contact or exposurePercutaneous exposureScratch, Puncture, Needle stickMucus membrane exposureEyes, Mouth, open cut
30 BSL-2 Practices Limited access to lab when work in progress Daily decontaminationMechanical pipettingLabcoat, safety glasses and gloves requiredRed bag & sharps containers required
31 BSL-2 Practices (con’t) Biohaz. Sign posted at entrance to labLabel all equipment (incubators, freezers, etc.)TC room – negative air flowDocumented trainingBaseline serology or pre-vaccination may be required
32 Risk Group 3 Agents Human Immunodeficiency Virus Mycobacterium tuberculosisCoxiella burnetii
33 Biosafety Level 3 Working in High Containment Biosafety Level-3 (BSL-3 or ABSL-3)Indigenous or exotic agentsAerosol transmissionSerious health effectsTreatment may or may not exist
34 BSL-3 Practices Public access NOT permitted Daily decontamination after spill and upon completion of experimentAutoclave required and waste is disposed at the end of dayRequired foot activated handwashing sink and controlsNo sharps unless absolutely necessary
35 BSL-3 Practices (con’t) Aerosol minimization procedures requiredWrap around disposable clothing is required. Specialized equipment may be required depending upon proceduresBiohaz. Signs and labels postedAir flow from low hazard to high hazard“Pressure Mapping”
36 BSL-3 Practices (con’t) Bench top work not permittedDocumented training and personnel competency certification (for BSL-3 procedures)Baseline serologySpills – report immediately and treat accordinglyVaccinations/post exposure protocols and SOP’s, Biosafety Manual, Biosafety Officer
38 Biosafety Level-4 Working in High Containment Builds on BSL-3/ ABSL-3 practicesMaximum containment facilitiesPressurized Containment SuiteBSL-3 + Class III Biosafety CabinetChemical decontamination showersLiquid effluent collection / decontaminationNo BSL-4 labs exist at UCSD
40 Biosafety Concepts Working in High Containment Biosafety Level-4 (BSL-4 or ABSL-4)Dangerous/exotic agentsLife threatening diseaseAerosol transmissionAgents of unknown riskof transmission or health affectsNo known treatment
41 Animal Biosafety Level-4 Working in High Containment
42 General Good Lab Technique Hygienic PracticesNo Smoking, Eating, Applying cosmetics, lip balm, contactsWash hands after proceduresDecontaminate lab bench before and after work
43 General Operational Practices Proper attireMinimum – lab coat, safety glasses, glovesPlan your workKnow in advance what you are working withRead available resources (MSDS)
44 Animal Containment Points CDCCDC & UCSDCourtesy of Paul Vinson, CDC
45 Discussion # 2Based on what you know about Biosafety Levels, Practices and Operational Controls, what are some discussion issues for conducting Biohazard risk assessments?How do you approach risks when addressing a particular organism?
48 Addressing Risk Assessments What is the organism?Is it Wild-type, attenuated, irradiated, or chemically treated? Look at kill data or kill curves.What is the max. concentration, volume, infectious dose?What is the work space like?Aerosolizing procedures? How do they contain their aerosols?
49 Risk Assessment, con’tTom PughAre personnel trained? Do personnel understand the organism, infectious dose and symptoms?What are their experimental procedures?Will they be transporting the material? Shipping intra, inter-state or international?Are they doing tissue culture?Do they have adequate containment equipment?
50 Risk Assessment, Con’tAre they doing this work in-vivo? Have you consulted and discussed this with the Vets and IACUC to determine special needs and housing?Waste issues addressed?Pregnancy issues with the organisms?
51 Risk Assessment, con’t Do they share their Tissue Culture room? Do they have more than 1 Biosafety Cabinet?Occupational Health informed and set up to receive patient or offer counseling?
52 Accidental SpillsEvacuate area, alert personnel and cordon off so that aerosols may settleDon PPE; Cover with paper towels and apply bleach (1 part bleach : 9 parts waterAllow 15 – 20 min contact timeWipe up working towards centerUse tongs if broken glass is involvedIs Recombinant DNA involved?
53 First Aid Measures Splash to Eye or Needlestick Injury Rinse thoroughly for 15 minutes at the eyewash or sinkCall Occupational Medicine Call EH&S to report exposure –
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