4 What is bioterrorismThe US president's 2006 budget requests more than $4 billion for biodefense activities at the Department of Health and Human Services (HHS).Within budget, approximately $1.7 billion will fund medical research and product development at the National Institute of Allergy and Infectious Diseases (NIAID), includingVaccines to immunize the public against diseases caused by bioterrorism agentsDiagnostic tests to help first responders and other medical personnel rapidly detect exposure and provide treatmentTherapies to help patients exposed to bioterrorism agents regain their health
5 Biosafety Personal protection Work place practices Administrative EngineeringAll play an equal role in protecting workers from occupational exposures in laboratories
6 Biosafety Personal protection Work place practices PPE, respirotors, gloves, booties, eye and ear protectionsWork place practicesEntry and exit practices, decontamination, emergency procedure, handle sharp.
8 CDC Bioterrorism Agents Category ACategory BCategory C
9 Category A Diseases/Agents can be easily disseminated or transmitted from person to person;result in high mortality rates and have the potential for major public health impact;might cause public panic and social disruption; andrequire special action for public health preparedness.
10 Category B Diseases/Agents Second highest priority agents include those thatare moderately easy to disseminate;result in moderate morbidity rates and low mortality rates; andrequire specific enhancements of CDC's diagnostic capacity and enhanced disease surveillance.
11 Category C Diseases/Agents Third highest priority agents include emerging pathogens that could be engineered for mass dissemination in the future because ofavailability;ease of production and dissemination; andpotential for high morbidity and mortality rates and major health impact.
16 The US National Institute of Allergy and Infectious Diseases (NIAID)
17 The US National Institute of Allergy and Infectious Diseases (NIAID)
18 Helen Quill & Maria Giovanni, Nature Immunology, 2004
19 Microbiological laboratories Microbiological laboratories are special, often unique work environments that may pose identifiable infectious disease risks to persons in or near them.
20 ContainmentPrimary containment, the protection of personnel and the immediate laboratory environment from exposure to infectious agents, is provided by both good microbiological technique and the use of appropriate safety equipment.Secondary containment, the protection of the environment external to the laboratory from exposure to infectious materials,
21 Laboratory Practice and Technique Develop or adopt a biosafety or operations manual that identifies the hazards that will or may be encountered, and that specifies practices and procedures designed to minimize or eliminate exposures to these hazards.
22 Principles of Biosafety Laboratory personnelSafety practices, and techniques must be supplementedAppropriate facility design and engineering featuresSafety equipment, and management practices.
23 Safety Equipment (Primary Barriers) Biological safety cabinets (BSCs)Enclosed containers,Engineering controls designed to remove or minimize exposures to hazardous biological materials.
24 The biological safety cabinet (BSC) Device used to provide containment of infectious splashes or aerosols.Three types of biological safety cabinets (Class I, II, III) used in microbiological laboratories.Open-fronted Class I and Class II biological safety cabinets are primary barriers which offer significant levels of protection to laboratory personnel.Safety centrifuge cup, an enclosed container designed to prevent aerosols.
28 Personal Protection gloves, coats, gowns, shoe covers, boots, respirators,face shields,safety glasses, or goggles.
29 Facility Design and Construction (Secondary Barriers) Depend on the risk of transmission of specific agents.Separation of the laboratory work area from public access, availability of a decontamination facility (e.g., autoclave), and handwashing facilities.If an infectious aerosol is present, higher levels of primary containment and multiple secondary barriers may become necessary to prevent infectious agents from escaping into the environment.
37 Biosafety Level 2 Definition Work with agents of moderate potential hazard to personnel and the environment. Agents associated with human disease which is rarely serious and for which preventative or therapeutic interventions are often available.
38 Biosafety Level 2 Requirements All requirements for BSL1 plus: Training to handle organismsPersonal protective equipment (PPE)- lab coats, eyewear, and glovesMinimize aerosol generationBiohazard sign posted on door
39 Biosafety Level 2 Examples Pathogenic E. coli Salmonella sp. Streptococcus sp.Hepatitis B and C Viruses
40 Biosafety Level 3 Definition Agents associated with serious or lethal disease for which preventative or therapeutic intervention may be available. High individual risk, low community risk. Often risk is by inhalation route.
41 Biosafety Level 3 Requirements All requirements for BSL2 plus: Limited lab access2 doors in series to access labAble to wash entire labSpecial exhaust ventilation- not recirculated
55 Biosafety Level 4 Definition Agents likely to cause serious or lethal disease for which preventative or therapeutic intervention is usually NOT available. Dangerous or exotic agents.
56 Biosafety Level 4 Requirements All requirements for BSL3 plus: Class III biosafety cabinet or positive pressure suitsShower/change rooms - clothing autoclaved before launderingAir locks
57 Biosafety Level 4 Examples No Bacteria or Fungi Ebola Virus Monkey B VirusMarburg Virus
58 Safe Work PracticesWash hands after work; before leaving lab; when removing glovesNo eating, drinking, applying cosmetics, handling contact lenses in labMaintain labs in clean, orderly fashionLimit access to lab when work with organisms is in progressUse good microbiological techniquesNo mouth pipettingWhen possible use plastic instead of glass
59 Personal Protective Equipment Contaminated personal protective equipment (PPE) should remain in the work area- do not wear in any "clean" areasPPE that is contaminated must be discarded as infectious waste or disinfected prior to routine laundering
60 Laminar Flow Equipment Biosafety cabinets (BSCs)Contain infectious agents to protect personnel and the environmentLaminar flow clean benches (LFBs)Non-hazardous work onlyProtect work from contamination
61 Laminar Flow Benches DO NOT protect personnel or the environment Discharges HEPA filtered air across work surface toward userDO NOT use biohazardous, radioactive, chemical, toxic, mutagenic or carcinogenic agents in a LFBShould have a pink sticker stating this
63 Biosafety Cabinets 3 Classes All exhaust is HEPA filtered before leaving cabinetClass I- do not protect the work from contaminationAir entering cabinet is not filtered
64 Biosafety Cabinets Class II- 4 types- A, B1, B2, B3 Each type recirculates different amount of airSome are hard-ducted, some exhaust into roomClass IIITotally enclosed, ventilated cabinetsWork through portals with attached gloves
65 Use of EquipmentMinimize airflow disturbances (moving in/out of cabinet, people walking by, opening doors, blocking grilles with equipment)Wipe down surfaces with alcohol before and after work. Let run 10 minutes to clean itMinimize use of flames in cabinetTry not to use chemicals in cabinets- only use in Class I or II hoods that are ducted- contact DOHSDo not store excess equipment in cabinet
66 Centrifuge SafetyAerosol generation from broken tubes, opening tubes, decanting supernatant, resuspending materialsBalance loadUse sealed tubes, safety buckets or rotors when possibleWhen possible fill & open centrifuge tubes or buckets in BSC
67 Centrifuge Accidents If a tube breaks- Turn off centrifuge Leave the labCall OfficersIf you find a tube broke when you open the centrifuge-Close the lid
68 Shipping/ Transporting Who regulate transportation of biologicals????Includes infectious materials, diagnostic specimens, dry iceProgram being developed- to include mandatory training
70 DisinfectionAll equipment, environmental, and working surfaces must be cleaned and decontaminated after work with infectious materialsBroken glass or sharps shall be cleaned up using mechanical means- NOT by handDifferent levels of cleaning- decontamination, sanitization, disinfection, sterilization
71 Decontamination Use appropriate disinfectant for agents in use Follow instructions, paying attention to dilution, shelf life, and contact time to assure effective killCare must be used to ensure mixing of incompatible materials does not occur
72 Infectious Waste What is infectious waste Procedures for disposal Solid wasteLiquid wasteSharpsAutoclaving wasteContainers and pick-upsMixed waste
74 BSL2 Spills Small spills- Wipe up with disinfectant-soaked paper towel then clean surface with suitable disinfectantLarge BSL2 Spill in BSCKeep cabinet running during and 10 minutes after cleanupDon PPECover spill with disinfectant- let set- wipe upClean catch basin under work surface if spill ran thereRemove PPE when done and washLeave the lab, close door, post lab off-limitsDecon people if necessaryAfter 30 minutes, put on PPE and assemble supplies
79 Bacteria Virus Parasite Fungus Agents Lab Animals B. anthracis, Bordetella pertussis, N. gonorrhoeae, N. meningitidis, B. pseudomallei, L. pneumophila, S. typhi, Y. pestisBrucella sp, F. tularensis,Campylobacter spp, C. diphtheriae, E. coli (cytotixin), H. pyroli, L. interrogans, L. monocytogenes, M. leprae, Salmonella spp., Shigella spp., T. pallidum, V. choleraM. tuberculosisBSL 2, BSL 3BSL 2BSL 3VirusPrion, RetrovirusRabies virusRickettsial agents, LCM, Hepatitis B, C, DHerpes virus, Hepatitis A, E, influenza, Poliovirus, Poxvirus,Other zoonosis virusParasiteAll pathogenic parasitesFungusBlastomyces dermatitidis, C.neoformans, Histoplasma capsulatum, Sporothrix schenckii, Miscellaneous MoldsCoccidioides immitis
86 Biosafety Agents Practices Safety Equipment Facilities Level BSL-1 not generally associated with disease· Good microbiological practice Hand washing No eating, drinkingPipeting devices- mouth pipeting is prohibitedBSL-2These agents are associated with human disease· Limited lab access. Most work may be performed on a bench top Biohazard warning. Biosafety manual defining any needed waste decontamination or medical surveillance policies· Class I or II Biological Safety Cabinets. Lab coats, gloves, face protection, as needed· Open bench-top sink for hand washing is requiredAutoclave
87 Agents Practices Safety Equipment Facilities LevelAgentsPracticesSafety EquipmentFacilitiesBSL-3· associated with human disease and cause illness by spreading through the airdiseases that may have serious or lethalBSL-2 practice plus· Controlled access· Decontamination of all wasteDecontamination of lab clothing before laundering· Class I or II Biological Safety Cabinets (BSCs) or other physical containment devicesProtective lab clothing, gloves, respiratory protection as neededBSL-2 plus· Physical separation from access corridors· Self-closing, double-door access· Exhaust air is not recirculated· Negative airflow into laboratoryDesign includes back up/redundant systemsBSL-4· BSL-3 or have an unknown cause of transmissionCause diseases that are usually life-threateningBSL-3 practices plus· Clothing change before entering· Shower on exitAll material decontaminated on exit from facilityAll procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive- pressure personnel suitBSL-3 plus· Separate building or isolated zone· Dedicated supply and exhaust, vacuum, and decontamination systems· Design includes back-up/redundant systems
88 Thanks you Biosafety in the Microbiological and Biomedical Laboratory More informationBiosafety in the Microbiological and Biomedical Laboratory
94 Acknowledgements T2 (Thailand Tropical Diseases Research Program). TRF (Thailand Research Fund).NSTDAONSITE, Emory UniversityFaculty of Science, MUThe Virology Association of ThailandFaculty of Medicine, KKU.