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  • Safety Survival SkillsII. Laboratory Safety

    A Primer on Safe Laboratory Practiceand Emergency Response for CDC Workers

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    A Primer on Safe Laboratory Practiceand Emergency Response for CDC Workers

    Developed by:Richard J. Green, MSc

    Office of Health and SafetyTraining Manager

    Centers for Disease Control and Prevention1600 Clifton Rd., F05

    Atlanta, GA 30333


  • Office of Health and SafetyDirector: 404-639-2453

    Training Activity: 404-639-2145/6Laboratory Safety Branch : 404-639-3235

    Environmental, Health & Safety Branch: 404-639-3142

    OHS Web SiteOffice of Health and Safety Information System

    Emergency ResponseAfter-hours Hotline: 404-639-4444 or 770-488-4444

  • Contents Page

    1. Laboratory Hazards 1Lab Hazards Routes of Exposure Lab AccidentsDirty Dozen Chemicals Laboratory Workers Risk Assessments

    2. Basic Biosafety 3Biosafety Levels BSL2 Standard Practices BSCs CentrifugesWaste Disposal Emergencies

    3. Basic Chemical Safety 11Golden Rules Standard Practices Fume Hoods Waste DisposalPacking and Moving Chemicals ImP2ACT EmergenciesRadiation Safety

    4. AppendicesA. Biological Waste Disposal Chart 25B. Required Door Signs 27C. Hazardous Waste Forms 29D. CDC Packaging/Shipping Information 31E. IATA Shipping Regulation Changes 35F. Export Controls for Biologicals, Chemicals, and 37

    Related Technical Data and EquipmentG. Occupational Health Clinic (OHC) Services 43H. Guidelines for Children in the Workplace in NCID 49 I. Guidelines for Filming in CDC Labs 55 J. Laboratory Safety Final Exam 57

    Safety Survival Skills


  • Safety Survival Skills

  • Safety Survival Skills

    Laboratory Hazards

    The laboratory environment is a hazardous place to work. Walkthrough any laboratory door and you are confronted with a widearray of chemicals, biologics, and instrumentation. Nearly everycommon laboratory technique, practice, or procedure carriessome risk of exposure or mechanical injury. Appreciating whatthese risks are and how to work safely with them is the focus ofthis manual.

    Lab Hazards Biological Chemical Ionizing Radiation Physical

    The Most Hazardous Dirty Dozen Chemicals* Organic azides Perchlorate salts of organic, organometallic, and

    inorganic complexes Diethyl ethers Lithium aluminum hydride Sodium, potassium Potassium metal Sodium-benzophenone ketyl still pots Palladium on carbon Heat generated from exothermic reactions Ethers with alpha hydrogen atoms Carbon monoxide Organic peroxides

    *Laboratorians should consult with OHS before working with these chemicals.

    Laboratorians should never assume that they are performing alltasks in a safe and correct manner just because they havenever had a laboratory accident. Both the hazard and the routeof transmission should be known before beginning any labora-tory procedure. Unfortunately, shortcuts are taken, materials areviewed as non-hazardous the longer they are worked with, andequipment is assumed to be functioning properly when they areturned on.

    Laboratory Hazards1

    Safety Incident

    I was cleaning paper out ofdrain in the sink with gloves onwhen I was pricked by a piece

    of glass on the left thumb.

    Safety Incident

    Laboratorian is heating ethylether in a beaker on a hot platein a fume hood. As more liquid

    ether is being added to thebeaker, the hot plate thermo-stat engages and ignites the

    ether fumes.

  • Safety Survival Skills

    Most Frequent Lab Accidents Splashes/Spills Needlesticks/Cuts Back injuries Explosions Fires Exothermic reactions Toxic fumes Mouth pipetting Animal bites/Scratches

    Recent studies have shown that there are definite differences inhow various people view safety.

    Laboratory Workers Unsafe Workers

    have low opinion of safety programs take excessive risks work too fast less aware of infectious risks young males (17-24 years of age)

    Safe Workers know and adhere to safety regulations have defensive work habits recognize potentially hazardous situations women and older employees (45-64 years of age)

    Risk AssessmentsPrior to beginning any laboratory procedure, laboratoriansshould gather all available information concerning the materialsthey will be working with and perform a risk assessment of theirmethods.

    1. Identify all hazardous materials (microbiologicals andchemicals) to be used and the circumstances for theiruse.

    2. Consult information resources (BMBL, MSDS).3. Evaluate the biosafety level and/or type of toxicity of

    the material.4. Consider possible routes of exposure.5. Evaluate quantitative information on toxicity.6. Select appropriate procedures to minimize exposure.7. Prepare for contingencies.

    Laboratory Hazards2

    Safety Incident

    I was working with infantmice infected with Strep.

    pneumoniae, trying to drawblood from the tail vein with amonolet lancet. I pushed downtoo hard and it went through

    the tail and into thenuckle of my leftmiddle finger.

    Safety Statistic

    In 1999, 69 CDClaboratorians were treatedat the Occupational Health

    Clinic for a variety oflaboratory incidents.

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    Basic Biosafety

    Microbiological laboratories are special, often unique workenvironments that may pose identifiable infectious disease risksto persons in or near them. Infections have been contracted inthe laboratory throughout the history of microbiology. Publishedreports around the turn of the century described laboratory-associated cases of typhoid, cholera, glanders, brucellosis, andtetanus.

    The term containment is used in describing safe methods formanaging infectious materials in the laboratory environmentwhere they are being handled or maintained. The purpose ofcontainment is to reduce or eliminate exposure of laboratoryworkers, other persons, and the outside environment to poten-tially hazardous agents. Primary containment, the protection ofpersonnel and the immediate laboratory environment fromexposure to infectious agents, is provided by both good micro-biological technique and the use of appropriate safety equip-ment. The use of vaccines may provide an increased level ofpersonal protection. Secondary containment, the protection ofthe environment external to the laboratory from exposure toinfectious materials, is provided by a combination of facilitydesign and operational practices. Therefore, the three elementsof containment include laboratory practice and technique, safetyequipment, and facility design. The risk assessment of the workto be done with a specific agent will determine the appropriatecombination of these elements.

    Biosafety Level (BSL)The recommended biosafety level(s) for the organisms repre-sent those conditions under which the agent ordinarily can besafely handled. The laboratory director is specifically and prima-rily responsible for assessing the risks and appropriately apply-ing the recommended biosafety levels. When specific informa-tion is available to suggest that virulence, pathogenicity, antibi-otic resistance patterns, vaccine and treatment availability, orother factors are significantly altered, more (or less) stringentpractices may be specified.

    BSL1 - work with agents not known to cause diseasein healthy adults; standard microbiological practicesSMP) apply; no safety equipment required; sinks re-quired.


    The Biohazard symbolmust be affixed to

    any container or equip-ment used to store ortransport potentiallyinfectious materials.

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    BSL2 - work with agents associated with human disease;SMP apply plus limited access, biohazard signs, sharpsprecautions, and biosafety manual required; BSC usedfor aerosol/splash generating operations; lab coats,gloves, face protection required; contaminated waste isautoclaved.

    BSL3 - work with indigenous /exotic agents which mayhave serious or lethal consequences and with potentialfor aerosol transmission; BSL2 practices plus controlledaccess; decontamination of all waste and lab clothingbefore laundering; determination of baseline serums;BSC used for all specimen manipulations; respiratoryprotection used as needed; physical separation fromaccess corridors; double door access; negative airflowinto lab; exhaust air not recirculated.

    BSL4 - work with dangerous/exotic agents of life threat-ening nature or unknown risk of transmission; BSL3practices plus clothing change before entering lab;shower required for exit; all materials are decontami-nated on exit; positive pressure personnel suit requiredfor entry; separated/isolated building; dedicated airsupply/exhaust and decon systems.

    Each of the four biosafety levels (BSLs) consist of combinationsof laboratory practices and techniques, safety equipment, andlaboratory facilities. Each combination is specifically appropriatefor the operations performed, the documented or suspectedroutes of transmission of the infectious agents, and the labora-tory function or activity. However, common to all four biosafetylevels are the Standard Practices which remain the same fromBSL1 to BSL4.

    Standard Practices1. Access to lab is limited or restricted by the lab director

    when work with infectious agents is in progress.2. Persons wash their hands after handling viable mate-

    rial and animals, after removing gloves, and beforeleaving lab.

    3. Eating, drinking, smoking, handling contact lenses,and applying cosmetics are not permi