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.
Basic Principles of Biosafety
Basic Principles of BiosafetyBasic Principles of BiosafetyMiguel Martin N. Moreno II, MDMiguel Martin N. Moreno II, MDScientist, SLMC, QC - RBDScientist, SLMC, QC - RBDFounding Secretary, PhBBA NationalFounding Secretary, PhBBA National
What is Biosafety? Safety from exposure toInfectious AgentsSmallpox
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 ascientist may be connected to bacteria he researched that causes the plague.The university said Saturday that its researcher studied the genetics of harmful bacteria includingYersinia pestis, which causes the illness. He died Sept. 13. His name and age havent beenreleasedThe medical center says the bacteria he worked with was a weakened strain that isnt known to causeillness in healthy adults. The strain was approved by the Centers for Disease Control andPrevention for laboratory studies.An autopsy found no obvious cause of death but did find the presence of the bacteria. More tests areplanned. No other illnesses have been reported.
Biosafety in Various DisciplinesBiosafety is related to several fields ECOLOGY: referring to importedlife forms not indigenous to theregion (Reggie the alligator) AGRICULTURE: reducing therisk of alien viral or transgenicgenes, or prions such asBSE/"MadCow“; reducing the riskof food bacterial contamination MEDICINE: referring toorgans or tissues from biologicalorigin, or genetic therapyproducts, virus; levels of labcontainment protocols BSL-1, 2,3, 4 in rising order of danger CHEMISTRY: i.e., nitrates inwater, PCB levels affectingfertility EXOBIOLOGY: i.e., NASAspolicy for containing alienmicrobes that may exist on spacesamples. (Classified info.)
Biosafety in Academic Research Research Universities:Promoting safe laboratorypractices, and procedures;proper use of containmentequipment and facilities;provides advice onlaboratory design and riskassesment of experimentsinvolving infectious agents,rDNA in-vitro and in-vivo.Bottom Line: Risk & Containment
Biohazard Symbol Charles Baldwin atNational CancerInstitute at NIH. Symbol to be“memorable butmeaningless” so it couldbe learned. Blaze orange – mostvisible under harshconditions
Biosafety Issues (con’t.) Respiratory Protection Bioterrorism and Select agents Mold and indoor air quality Occupational safety and health in the useof research animals Biohazards used in animal models
Biohazardous Materials Viruses Bacteria Fungi Chlamydiae/Rickettsiae Prions Recombinant DNA
Biohazardous materials Transgenic Plants, Animals and Insects
Biohazardous Materials Human and Primate Cells, Tissues, andBody Fluids Brain Tissue from Demented Patients Viral Vectors Replication deficient viruses
Biosafety In Microbiologicaland Biomedical LaboratoriesBMBLCDC/NIH PublicationSafety “Guidelines”Regulations of InstitutionreceivesNIH fundingCode of Practice and “Gold”Standard in IndustryGold StandardClinical & Research Lab.Lab. Animal FacilitiesBiosafety Conceptshttp://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
Canadian Laboratory Biosafety Guidelineshttp://www.phac-aspc.gc.ca/publicat/lbg-ldmbl-04/pdf/lbg_2004_e.pdf
Philippines Manual of Standards on NationalLaboratory Biosafety & Biosecurity by the Task Force/TWG on thePreparation of Standards on NationalLaboratory Biosafety & Biosecurity asper DP) No. 2006-2500 DOH, National Center for Health FacilityDevelopment, 17 March 2009
Biosafety ConceptsPrinciples of Biosafety Practice and Procedures Standard Practices Special Practices & Considerations Safety Equipment Facility Design and Construction Increasing levels of protection
Principles of BiosafetyBiosafety Levels 1-4 (BSL) Increasing levels of employee and environmental protection Guidelines for working safely in research & medical laboratoryfacilitiesAnimal Biosafety Levels 1- 4 (ABSL) Laboratory animal facilities Animal models that support research Guidelines for working safely in animal research facilities
Biosafety Concepts(1) Standard Microbiological Practices Most important concept / Strict adherence Aware of potential hazard Trained & proficient in techniques Supervisors responsible for: Appropriate Laboratory facilities Personnel & Training Special practices & precautions Occupational Health Programs
(2) Safety Equipment Primary Containment Barrier Minimize exposure to hazard Prevent contact / Contain aerosols Engineering controls/ equipment Personal Protective Equipment (PPE) Gloves, gowns, Respirator, Face shield, Booties Biological Safety Cabinets Covered or ventilated animal cage systemsBiosafety Concepts
(3) Facility Design andConstruction Secondary Barrier/ Engineeringcontrols Contributes to worker protection Protects outside the laboratory Environment & Neighborhood Ex. Building & Lab design,Ventilation, Autoclaves, Cagewash facilities, etc.Biosafety Concepts
Biosafety Level-1Concepts of BiosafetyBiosafety Level-1 (BSL-1 or ABSL-1) Well characterized agents Agents not known to cause disease (in healthy humanadults; now healthy immunocompetent adults) Prophylactic treatment available Open bench procedures Animals in open cage system or open environment(outdoors) Good laboratory practices
BSL-1 Practices Bench-top work allowed Daily Decontamination Manual pipetting Required Handwashing Red bag waste Bio cabinet not required(unless creating aerosols) 2˚ containment
Risk Group 2 Agents Human or PrimateCells Herpes Simplex Virus ReplicationIncompetentAttenuated HumanImmunodeficiencyVirus Patient specimens
BSL-2 PracticesConcepts of BiosafetyPractices & Procedures Agents associated w/ human disease Treatment for disease available Agent poses moderate hazard topersonnel and environment Direct contact or exposure Percutaneous exposure Scratch, Puncture, Needle stick Mucus membrane exposure Eyes, Mouth, open cut
BSL-2 Practices Limited access to labwhen work in progress Daily decontamination Mechanical pipetting Lab.coat, safety glassesand gloves required Red bag & sharpscontainers required
BSL-2 Practices (con’t) Biohaz. Sign posted atentrance to lab Label all equipment(incubators, freezers, etc.) TC room – negative air flow Documented training Baseline serology or pre-vaccination may be required
Risk Group 3 Agents HumanImmunodeficiencyVirus Mycobacteriumtuberculosis Coxiella burnetii
Biosafety Level 3Working in High ContainmentBiosafety Level-3 (BSL-3 orABSL-3) Indigenous or exotic agents Aerosol transmission Serious health effects Treatment may or may not exist
BSL-3 Practices Public access NOT permitted Daily decontamination after spill andupon completion of experiment Autoclave required and waste isdisposed at the end of day Required foot activated handwashingsink and controls No sharps unless absolutely necessary
BSL-3 Practices (con’t) Aerosol minimization procedures required Wrap around disposable clothing isrequired. Specialized equipment may berequired depending upon procedures Biohaz. Signs and labels posted Air flow from low hazard to high hazard“Pressure Mapping”
BSL-3 Practices (con’t) Bench top work not permitted Documented training and personnel competencycertification (for BSL-3 procedures) Baseline serology Spills – report immediately and treat accordingly Vaccinations/post exposure protocols and SOP’s,Biosafety Manual, Biosafety Officer
Biosafety Level-4Working in High ContainmentBiosafety Level-4 Builds on BSL-3/ ABSL-3 practices Maximum containment facilities Pressurized Containment Suite BSL-3 + Class III Biosafety Cabinet Chemical decontamination showers Liquid effluent collection / decontamination
Biosafety ConceptsWorking in High ContainmentBiosafety Level-4 (BSL-4 or ABSL-4) Dangerous/exotic agents Life threatening disease Aerosol transmission Agents of unknown riskof transmission or health affects No known treatment
General Good Lab Technique Hygienic Practices No Smoking, Eating, Applying cosmetics, lipbalm, contacts Wash hands after procedures Decontaminate lab bench before and afterwork
General Operational Practices Proper attire Minimum – lab coat, safety glasses, gloves Plan your work Know in advance what you are working with Read available resources (MSDS)http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/index.html
Risk AssessmentIn-Vitro In-Vivo Human ClinicalTrial
Addressing Risk Assessments What is the organism? Is it Wild-type, attenuated, irradiated, orchemically treated? Look at kill data or killcurves. What is the max. concentration, volume,infectious dose? What is the work space like? Aerosolizing procedures? How do they containtheir aerosols?
Risk Assessment (con’t) Are personnel trained? Dopersonnel understand the organism,infectious dose and symptoms? What are their experimentalprocedures? Will they be transporting thematerial? Shipping intra, inter-stateor international? Are they doing tissue culture? Do they have adequate containmentequipment?
Risk Assessment (con’t) Are they doing thiswork in-vivo? Haveyou consulted anddiscussed this with theVets and IACUC todetermine special needsand housing? Waste issues addressed? Pregnancy issues withthe organisms?
Risk Assessment (con’t) Do they share theirTissue Culture room? Do they have more than1 Biosafety Cabinet? Occupational Healthinformed and set up toreceive patient or offercounseling?
Accidental Spills Evacuate area, alert personnel andcordon off so that aerosols maysettle Don PPE; Cover with papertowels and apply bleach (1 partbleach : 9 parts water Allow 15 – 20 min contact time Wipe up working towards center Use tongs if broken glass isinvolvedIs Recombinant DNA involved?
First Aid Measures Splash to Eye or Needlestick Injury Rinse thoroughly for 15 minutes at the eyewash orsink Call Occupational Medicine Call Employee Health Services
Be safe, Be Biosafe, Be alive!Maraming Salamat po!