Biosafety at MUSC IBC website IBC website Michael G. Schmidt, Ph.D., Chair, Biosafety Committee.

Biosafety at MUSCBiosafety at MUSCBiosafety at MUSCBiosafety at MUSC

IBC websiteIBC website

Michael G. Schmidt, Ph.D., Chair, Biosafety CommitteeMichael G. Schmidt, Ph.D., Chair, Biosafety Committee

Biosafety at MUSCBiosafety at MUSC Learning ObjectivesLearning Objectives

1.1. Understand the role and responsibilities of the PI with Understand the role and responsibilities of the PI with respect to Biosafety and Animalsrespect to Biosafety and Animals

2.2. Understand the underlying principles of biosafety as Understand the underlying principles of biosafety as they relate to conducting safe and effective science they relate to conducting safe and effective science requiring the use of biohazardous materials on campusrequiring the use of biohazardous materials on campus

3.3. Be able to locate resources that will facilitate the Be able to locate resources that will facilitate the determination of the relative biosafety risk associated determination of the relative biosafety risk associated with a planned experimentwith a planned experiment

4.4. Recognize the significance and need for conducting Recognize the significance and need for conducting experiments at different biosafety containment levelsexperiments at different biosafety containment levels

5.5. Understand the differences between BSL1 and BSL2Understand the differences between BSL1 and BSL2

Biosafety at MUSCBiosafety at MUSC Learning Objectives continued:Learning Objectives continued:

6.6. Recognize the role each piece of Recognize the role each piece of Personal Protective Personal Protective Equipment Equipment plays in safeguarding the health and welfare plays in safeguarding the health and welfare of the laboratorian and communityof the laboratorian and community

7.7. Be familiar with the proper use of Biological Safety Be familiar with the proper use of Biological Safety Cabinets (BSC)Cabinets (BSC)

8.8. Recognize the inherent risk with using centrifuges in Recognize the inherent risk with using centrifuges in the laboratory with a special emphasis on risks the laboratory with a special emphasis on risks associated with infectious agentsassociated with infectious agents

9.9. Understand the significance of good laboratory Understand the significance of good laboratory housekeepinghousekeeping

10.10. Be able to clean up a minor biological spill within a Be able to clean up a minor biological spill within a BSCBSC

11.11. Understand what is required of you during the bi-Understand what is required of you during the bi-annual Laboratory Biological Safety Inspection annual Laboratory Biological Safety Inspection


12.12. Understand the signficance of working with Understand the signficance of working with biohazardous Materials specifically the role of the biohazardous Materials specifically the role of the PI in informing the laboratorian of potential PI in informing the laboratorian of potential hazards in the workplace as well as the personal hazards in the workplace as well as the personal responsibility required of each employee and responsibility required of each employee and studentstudent

Biosafety at MUSCBiosafety at MUSC Unit 1Unit 1

The role and responsibilities of the PI with respect The role and responsibilities of the PI with respect to Biosafety and Animals at MUSCto Biosafety and Animals at MUSC

Biosafety in Biomedical Biosafety in Biomedical Laboratories at MUSCLaboratories at MUSCBiosafety in Biomedical Biosafety in Biomedical Laboratories at MUSCLaboratories at MUSC The role and responsibilities of the PI The role and responsibilities of the PI

with respect to Biosafety and Animalswith respect to Biosafety and Animals A full-time faculty or professional staff member A full-time faculty or professional staff member

with an advanced degree must agree to accept with an advanced degree must agree to accept responsibility for the project and serve as the responsibility for the project and serve as the PI. PI.

Student researchers, residents, and postdoctoral Student researchers, residents, and postdoctoral fellows cannot be identified as principal fellows cannot be identified as principal investigators.investigators.

The Principal Investigator (PI) is responsible The Principal Investigator (PI) is responsible for full compliance with the federal and state for full compliance with the federal and state regulations, NIH and CDC Guidelines, and regulations, NIH and CDC Guidelines, and institutional requirements for research institutional requirements for research involving biohazardous materials. involving biohazardous materials.

The PI is also responsible for ensuring that the The PI is also responsible for ensuring that the reporting requirements are fulfilled and will be reporting requirements are fulfilled and will be held accountable for any reporting lapses. held accountable for any reporting lapses.

DefinitionDefinitionDefinitionDefinition

BiohazardBiohazard An agent of biological origin that has An agent of biological origin that has

the capacity to produce deleterious the capacity to produce deleterious effects on humans, animals, plants and effects on humans, animals, plants and insects. These include microorganisms, insects. These include microorganisms, toxins and allergens derived from those toxins and allergens derived from those organisms; and allergens and toxins organisms; and allergens and toxins derived from insects, animals and plants.derived from insects, animals and plants.

http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf

Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories The role and responsibilities of the PI The role and responsibilities of the PI

with respect to Biosafety and Animalswith respect to Biosafety and Animals The Principal Investigator (PI) is responsible for full The Principal Investigator (PI) is responsible for full

compliance with the federal and state regulations, compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional NIH and CDC Guidelines, and institutional requirements for research involving biohazardous requirements for research involving biohazardous materials.materials.

http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm

PDF PDFhttp://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm

Biosafety at MUSCBiosafety at MUSC Questions for Unit 1Questions for Unit 1

The role and responsibilities of the PI with respect The role and responsibilities of the PI with respect to Biosafety and Animals at MUSCto Biosafety and Animals at MUSC


Principles of Biosafety as they relate to the safe Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous conduct of science requiring the use of biohazardous materials at MUSCmaterials at MUSC

DefinitionDefinitionDefinitionDefinition

BiohazardBiohazard An agent of biological origin that has An agent of biological origin that has

the capacity to produce deleterious the capacity to produce deleterious effects on humans, animals, plants and effects on humans, animals, plants and insects. These include microorganisms, insects. These include microorganisms, toxins and allergens derived from those toxins and allergens derived from those organisms; and allergens and toxins organisms; and allergens and toxins derived from insects, animals and plants.derived from insects, animals and plants.

Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories Principles of BiosafetyPrinciples of Biosafety

Containment Containment The purpose of containment is to reduce or The purpose of containment is to reduce or

eliminate exposure of:eliminate exposure of:• laboratory workers, laboratory workers, • other personsother persons• the environmentthe environment

Safe methods for managing infectious agents in Safe methods for managing infectious agents in the laboratorythe laboratory

EnvironmentEnvironment The location where the agent being handled, The location where the agent being handled,

maintained or stored.maintained or stored.

Infectious Agents and the lab workerInfectious Agents and the lab workerInfectious Agents and the lab workerInfectious Agents and the lab worker Modes of InfectionModes of Infection

Infections preceded by overt personal accidents, Infections preceded by overt personal accidents, which include:which include:

Inoculation Inoculation • resulting from pricking, jabbing or cutting the skin with resulting from pricking, jabbing or cutting the skin with

contaminated instruments such as:contaminated instruments such as:– hypodermic needles, scalpels and glassware; hypodermic needles, scalpels and glassware; – from animal bites or scratchesfrom animal bites or scratches

Ingestion Ingestion • resulting from mouth-pipetting, eating, drinking and smokingresulting from mouth-pipetting, eating, drinking and smoking

Splashing into the face and eyesSplashing into the face and eyes Spillage and direct contact Spillage and direct contact

Infectious Agents and the lab workerInfectious Agents and the lab worker

Infections not preceded by personal Infections not preceded by personal accidents:accidents: Aerosols, droplets and fomites. Aerosols, droplets and fomites.

These are speculated (from Pike’s 1976 data) to These are speculated (from Pike’s 1976 data) to be responsible for up to 82% percent of all be responsible for up to 82% percent of all laboratory-acquired infections. laboratory-acquired infections.

Aerosols are a cloud of very small liquid droplets Aerosols are a cloud of very small liquid droplets produced whenever energy is applied to a liquid, produced whenever energy is applied to a liquid, and such liquid is allowed to escape into the and such liquid is allowed to escape into the environment.environment.

• Centrifugation is a good source (plasmid Centrifugation is a good source (plasmid preps)preps)

Infectious Agents and the lab workerInfectious Agents and the lab worker

There are many regulations in place to forestall There are many regulations in place to forestall the problem of laboratory-acquired infections. the problem of laboratory-acquired infections. Responsibility for compliance with the regulations Responsibility for compliance with the regulations

Primarily to the Principal InvestigatorPrimarily to the Principal Investigator Secondarily, with the laboratory staff.Secondarily, with the laboratory staff.

In general a large number of organisms that would In general a large number of organisms that would ordinarily be innocuous can be infective in immune ordinarily be innocuous can be infective in immune compromised persons. compromised persons.

Therefore, additional and more stringent measures must Therefore, additional and more stringent measures must be established by the PI in an effort to prevent the be established by the PI in an effort to prevent the occurrence of lab-acquired infections in such individuals.occurrence of lab-acquired infections in such individuals.


Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories

Primary containmentPrimary containment Protection from exposure to infectious agents, is Protection from exposure to infectious agents, is

provided by both good microbiological technique provided by both good microbiological technique and the use of appropriate safety equipmentand the use of appropriate safety equipment..

SecondarySecondary containmentcontainment Protection of the environment external to the Protection of the environment external to the

laboratory from exposure to infectious materialslaboratory from exposure to infectious materials Facility design Facility design Operational practices Operational practices

Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories Three elements of containment include:Three elements of containment include:

1.1. Laboratory practice and technique, Laboratory practice and technique, 2.2. Safety equipmentSafety equipment

Bunsen burners vs. BactincineratorsBunsen burners vs. Bactincinerators Cages and other barriers Cages and other barriers

3.3. Facility designFacility design The risk assessment of the work to be done The risk assessment of the work to be done

with a specific agent will determine the with a specific agent will determine the appropriate combination of these elements.appropriate combination of these elements.

QuickTime™ and aTIFF (Uncompressed) decompressor

are needed to see this picture.

Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories Laboratory practice and techniqueLaboratory practice and technique

Standard microbiological practices and techniquesStandard microbiological practices and techniques Persons working with infectious agents or Persons working with infectious agents or

potentially infected materials must be aware of potentially infected materials must be aware of potential hazards, and must be trained and potential hazards, and must be trained and proficient in the practices and techniques required proficient in the practices and techniques required for handling such material safelyfor handling such material safely..

The director or person in charge of the laboratory is The director or person in charge of the laboratory is responsible for providing or arranging for responsible for providing or arranging for appropriate training of personnel.appropriate training of personnel.

Biosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical LaboratoriesBiosafety in Microbiological and Biosafety in Microbiological and Biomedical LaboratoriesBiomedical Laboratories A scientist trained and knowledgeable in appropriate laboratory A scientist trained and knowledgeable in appropriate laboratory

techniques, safety procedures, and hazards associated with techniques, safety procedures, and hazards associated with handling infectious agents must direct laboratory activities. handling infectious agents must direct laboratory activities.

Each laboratory should develop or adopt a biosafety or Each laboratory should develop or adopt a biosafety or operations manual which identifies the hazards that will or may operations manual which identifies the hazards that will or may be encountered, and which specifies practices and procedures be encountered, and which specifies practices and procedures designed to minimize or eliminate risks.designed to minimize or eliminate risks.

All personnel should be advised of special hazards and should All personnel should be advised of special hazards and should be required to read and to follow the required practices and be required to read and to follow the required practices and procedures. procedures. Each individual in the laboratory should acknowledge that they have Each individual in the laboratory should acknowledge that they have

been advised and briefed as to the laboratory’s plan and to the risks that been advised and briefed as to the laboratory’s plan and to the risks that they will be exposed.they will be exposed.

There is no such thing as zero risk! There is no such thing as zero risk!


Principles of Biosafety as they relate to the safe Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous conduct of science requiring the use of biohazardous materials at MUSCmaterials at MUSC


How to determine the relative biosafety risk How to determine the relative biosafety risk associated with a planned experiment associated with a planned experiment

Laboratory Containment Levels for Biological Laboratory Containment Levels for Biological Research Involving Potential BiohazardsResearch Involving Potential BiohazardsLaboratory Containment Levels for Biological Laboratory Containment Levels for Biological Research Involving Potential BiohazardsResearch Involving Potential Biohazards

Questions to Ask ?Questions to Ask ? Hazard LevelsHazard Levels

MSDS for MicrobesMSDS for Microbes

Standard Microbiological PracticesStandard Microbiological Practices Special PracticesSpecial Practices Containment EquipmentContainment Equipment

http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/







MSDS for MicrobesMSDS for Microbes• Issue AerosolIssue Aerosol

– 150 pfu is 150 pfu is infectious infectious intra-nasallyintra-nasally

• IncubationIncubation– 1-10 days1-10 days

• CommunicabilityCommunicability– YesYes




DisinfectantsDisinfectants• BleachBleach

Physical InactivationPhysical Inactivation Survival outside hostSurvival outside host

• Type 3 survival 10 Type 3 survival 10 days on paperdays on paper

• Type 2 survived 3-8 Type 2 survived 3-8 weeks on weeks on environmental environmental surfaces at room surfaces at room temperature.temperature.




10 case of lab 10 case of lab infectionsinfections

Special HazardsSpecial Hazards Contact with feces Contact with feces

from infected from infected animalsanimals

SpillsSpills DisposalDisposal





Standard Microbiological PracticesStandard Microbiological Practices Special PracticesSpecial Practices Containment EquipmentContainment Equipment



Infectious AgentsInfectious Agents Work may only be conducted with prior Work may only be conducted with prior

approval of the IBC regardless of the safety approval of the IBC regardless of the safety classification of the agentclassification of the agent

You must followYou must follow the requirements as the requirements as specified in the CDC/NIH specified in the CDC/NIH Biosafety in Microbiological and Biomedical LaboratorBiosafety in Microbiological and Biomedical Laboratories Manualies Manual

Containment requirements may be subject to Containment requirements may be subject to modification by the IBCmodification by the IBC


Define the riskDefine the risk Infectious Agents ListInfectious Agents List Experimental ProtocolExperimental Protocol

How big ?How big ? Aerosols ?Aerosols ? Animals ?Animals ?


SECTION II - SECTION II - Principles of Biosafety Principles of Biosafety Recommended Biosafety Levels for Recommended Biosafety Levels for

Infectious Agents in the LaboratoryInfectious Agents in the Laboratory Summary of Recommended Biosafety Levels Summary of Recommended Biosafety Levels

for Activities in Which Experimentally or for Activities in Which Experimentally or Naturally Infected Vertebrate Animals Are Naturally Infected Vertebrate Animals Are Used Used

http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s2.htm

http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm

PDF


Section III Laboratory- Biosafety Level Section III Laboratory- Biosafety Level CriteriaCriteria BSL 1, BSL 2, BSL 3, BSL 4 BSL 1, BSL 2, BSL 3, BSL 4 Comparison of Biological Safety Cabinets Comparison of Biological Safety Cabinets

Section IV-Vertebrate Animal Biosafety Level Section IV-Vertebrate Animal Biosafety Level CriteriaCriteria

ABSL 1, ABSL 1, ABSL 2, ABSL 2, ABSL 3,ABSL 3, ABSL 4 ABSL 4




SECTION V - Risk Assessment SECTION V - Risk Assessment SECTION VI- Recommended Biosafety SECTION VI- Recommended Biosafety

Levels For Infectious Agents and Infected Levels For Infectious Agents and Infected Animals Animals

Section VII- Section VII- Agent Summary StatementsAgent Summary Statements




Recombinant DNA, Gene Therapy Recombinant DNA, Gene Therapy and Transgenicsand TransgenicsRecombinant DNA, Gene Therapy Recombinant DNA, Gene Therapy and Transgenicsand Transgenics

Guidelines for Research Involving Guidelines for Research Involving Recombinant DNA MoleculesRecombinant DNA Molecules

Or in PDF at this siteOr in PDF at this site ScopeScope SafetySafety Experiments GovernedExperiments Governed Roles and ResponsibilitiesRoles and Responsibilities

http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Gdlnes_lnk_2002z.pdf


Guidelines for Research Involving Guidelines for Research Involving Recombinant DNA MoleculesRecombinant DNA Molecules

Classification of Human Etiologic Agents Classification of Human Etiologic Agents on the Basis of Hazardon the Basis of Hazard Appendix B-I. Risk Group 1 (RG1) AgentsAppendix B-I. Risk Group 1 (RG1) Agents Appendix B-II. Risk Group 2 (RG2) AgentsAppendix B-II. Risk Group 2 (RG2) Agents Appendix B-III. Risk Group 3 (RG3) AgentsAppendix B-III. Risk Group 3 (RG3) Agents Appendix B-IV. Risk Group 4 (RG4) AgentsAppendix B-IV. Risk Group 4 (RG4) Agents


Guidelines for Research Involving Recombinant DGuidelines for Research Involving Recombinant DNA MoleculesNA Molecules

Physical and Biological Containment for Recombinant DNA Physical and Biological Containment for Recombinant DNA Research Involving AnimalsResearch Involving Animals Appendix Q-I. General ConsiderationsAppendix Q-I. General Considerations

Appendix Q-I-A. Containment Levels Appendix Q-I-A. Containment Levels Appendix Q-I-B. Disposal of Animals (BL1-N through BL4-N)Appendix Q-I-B. Disposal of Animals (BL1-N through BL4-N) Appendix Q-II. Physical and Biological Containment LevelsAppendix Q-II. Physical and Biological Containment Levels Appendix Q-II-A. Biosafety Level 1 - Animals (BL1-N)Appendix Q-II-A. Biosafety Level 1 - Animals (BL1-N) Appendix Q-II-B. Biosafety Level 2 - Animals (BL2-N) Appendix Q-II-B. Biosafety Level 2 - Animals (BL2-N) Appendix Q-II-C. Biosafety Level 3 - Animals (BL3-N) Appendix Q-II-C. Biosafety Level 3 - Animals (BL3-N) Appendix Q-II-D. Biosafety Level 4 - Animals (BL4-N) Appendix Q-II-D. Biosafety Level 4 - Animals (BL4-N) Appendix Q-III. Footnotes and References for Appendix QAppendix Q-III. Footnotes and References for Appendix Q

http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm

Recombinant DNA, Gene Therapy and TransgenicsRecombinant DNA, Gene Therapy and TransgenicsRecombinant DNA, Gene Therapy and TransgenicsRecombinant DNA, Gene Therapy and Transgenics

Guidelines for Research Involving Recombinant DNA MoleculesGuidelines for Research Involving Recombinant DNA Molecules What’s Exempt?What’s Exempt?

See section III F (page 20 NIH Guide (April 02))See section III F (page 20 NIH Guide (April 02)) Those that are not in organisms or virusesThose that are not in organisms or viruses Those that consist entirely of DNA segments from a single non-Those that consist entirely of DNA segments from a single non-

chromosomal or viral DNA source, though one or more of the segments chromosomal or viral DNA source, though one or more of the segments may be syntheticmay be synthetic

Those that consist entirely of DNA from a prokaryotic host including its Those that consist entirely of DNA from a prokaryotic host including its indigenous plasmids or viruses when propagated in that host (or a closely indigenous plasmids or viruses when propagated in that host (or a closely related strain of the same species), or when transferred to another host by related strain of the same species), or when transferred to another host by well established physiological means. well established physiological means.

Those that consist entirely of DNA from a eukaryotic host including its Those that consist entirely of DNA from a eukaryotic host including its indigenous chloroplasts, mitochondria or plasmids (excluding viruses) indigenous chloroplasts, mitochondria or plasmids (excluding viruses) when propagated in that host.when propagated in that host.

Bottom line… PCR and gel running..Bottom line… PCR and gel running..

Biosafety at MUSCBiosafety at MUSC Questions Unit 3Questions Unit 3

How to determine the relative biosafety risk How to determine the relative biosafety risk associated with a planned experiment associated with a planned experiment


Specific Requirements for Biosafety Levels 1 to 3Specific Requirements for Biosafety Levels 1 to 3 PPEPPE Biological Safety Cabinets (BSCs)Biological Safety Cabinets (BSCs) Risks associated with centrifugation and proper protocolsRisks associated with centrifugation and proper protocols





For more information: http://www.washup.orghttp://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf



















Human Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell Culture Human blood, blood products, body fluids and Human blood, blood products, body fluids and

tissues are listed as potentially Hazardous tissues are listed as potentially Hazardous Biological MaterialsBiological Materials Biosafety Level 2 Practices and procedures MUST be Biosafety Level 2 Practices and procedures MUST be

followed when handlingfollowed when handling• BloodBlood• Blood ProductsBlood Products• Body fluidsBody fluids• TissuesTissues• Under no circumstance shall anyone work with cells derived Under no circumstance shall anyone work with cells derived

from themselves or from first degree relatives since the host from themselves or from first degree relatives since the host immune systems may not provide adequate protectionimmune systems may not provide adequate protection

• See Green book for greater detailSee Green book for greater detail

Human Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell Culture Cell CultureCell Culture

When a cell culture contains an etiologic When a cell culture contains an etiologic agent, oncogenic virus or amphotropic agent, oncogenic virus or amphotropic packaging system the cell line must be packaging system the cell line must be classified at the same level as that classified at the same level as that recommended for the agent.recommended for the agent.

Human Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell CultureHuman Tissue and Cell Culture Cell CultureCell Culture

The following cell lines are at Biosafety Level 2 or The following cell lines are at Biosafety Level 2 or higherhigher

All cell lines of human/primate origin All cell lines of human/primate origin Any cell lines derived from lymphoid or tumor tissue Any cell lines derived from lymphoid or tumor tissue All cell lines exposed to or transformed by any oncogenic virus All cell lines exposed to or transformed by any oncogenic virus All cell lines exposed to or transformed by amphotropic All cell lines exposed to or transformed by amphotropic

packaging systems packaging systems All clinical material (e.g., samples of human tissues and fluids All clinical material (e.g., samples of human tissues and fluids

obtained after surgical resection or autopsy) obtained after surgical resection or autopsy) All cell lines new to the laboratory (until proven to be free of all All cell lines new to the laboratory (until proven to be free of all

adventitious agents) adventitious agents) All mycoplasma-containing cell linesAll mycoplasma-containing cell lines

Personal Protective EquipmentPersonal Protective EquipmentPersonal Protective EquipmentPersonal Protective Equipment

PPE is used to protect you from contact PPE is used to protect you from contact with biohazardous materialswith biohazardous materials

Graphic credit: http://www-ehs.ucsd.edu/bio11.htm



Personal Protective EquipmentPersonal Protective EquipmentPersonal Protective EquipmentPersonal Protective Equipment

If the science you are If the science you are conducting requires you to conducting requires you to wear a N95 Respirator wear a N95 Respirator ((Mycobacterium Mycobacterium tuberculosistuberculosis) Occupational ) Occupational Health and Safety must Health and Safety must FIT-TEST you prior to FIT-TEST you prior to you starting work!you starting work!

















Laboratory HousekeepingLaboratory HousekeepingLaboratory HousekeepingLaboratory Housekeeping

Good housekeeping is essential forGood housekeeping is essential for Reducing risks Reducing risks Protecting the integrity of biological Protecting the integrity of biological

experimentsexperiments Keep the laboratory neat and free of Keep the laboratory neat and free of

clutter. clutter.


Specific Requirements for Biosafety Levels 1 to 3Specific Requirements for Biosafety Levels 1 to 3 PPEPPE Biological Safety Cabinets (BSCs)Biological Safety Cabinets (BSCs) Risks associated with centrifugation and proper protocolsRisks associated with centrifugation and proper protocols Cell Culture Cell Culture SpillsSpills


Biohazardous SpillsBiohazardous Spills

Biohazardous SpillsBiohazardous SpillsBiohazardous SpillsBiohazardous Spills Outside the Biosafety cabinetOutside the Biosafety cabinet

Clear area of all personnel. Clear area of all personnel. Wait at least 15 minutes for aerosol to settle Wait at least 15 minutes for aerosol to settle

before entering spill area. before entering spill area. Remove any contaminated clothing and Remove any contaminated clothing and

place in biohazard bag to be autoclaved. place in biohazard bag to be autoclaved. Put on a disposable gown, safety glasses and Put on a disposable gown, safety glasses and

gloves. gloves.

Biohazardous SpillsBiohazardous SpillsBiohazardous SpillsBiohazardous Spills Outside the Biosafety cabinetOutside the Biosafety cabinet

Initiate cleanup with disinfectant as follows: Initiate cleanup with disinfectant as follows: 1.1. Place dry paper towel on spill (to absorb liquids); then layer a Place dry paper towel on spill (to absorb liquids); then layer a

second set of disinfectant soaked paper towels over the spill. second set of disinfectant soaked paper towels over the spill. 2.2. Encircle the spill with additional disinfectant being careful to Encircle the spill with additional disinfectant being careful to

minimize aerosolization while assuring adequate contact. minimize aerosolization while assuring adequate contact. 3.3. Decontaminate all items within spill the area. Decontaminate all items within spill the area. 4.4. Allow minimum of 15 minutes contact time to ensure germicidal Allow minimum of 15 minutes contact time to ensure germicidal

action of disinfectant. action of disinfectant. 5.5. Wipe equipment with appropriate disinfectant. Wipe equipment with appropriate disinfectant. 6.6. Discard contaminated disposable materials using appropriate Discard contaminated disposable materials using appropriate

biohazardous waste disposal procedures (e.g., autoclave or BFI). biohazardous waste disposal procedures (e.g., autoclave or BFI). 7.7. Disinfect reusable items.Disinfect reusable items.

Biohazardous SpillsBiohazardous SpillsBiohazardous SpillsBiohazardous Spills Inside the Biosafety Cabinet (BSC)Inside the Biosafety Cabinet (BSC)

Wait at least five minutes to allow the BSC to contain Wait at least five minutes to allow the BSC to contain aerosols. aerosols.

Wear lab coat, safety glasses and gloves during cleanup. Wear lab coat, safety glasses and gloves during cleanup. Allow cabinet to run during cleanup. Allow cabinet to run during cleanup. Apply disinfectant and allow a minimum of 15 minutes Apply disinfectant and allow a minimum of 15 minutes

contact time. contact time. Wipe up spillage with disposable disinfectant-soaked Wipe up spillage with disposable disinfectant-soaked

paper towel.paper towel. Place used cleanup items in the appropriate biohazard Place used cleanup items in the appropriate biohazard

waste containerwaste container


Wipe the walls, work surface and any equipment in Wipe the walls, work surface and any equipment in the cabinet with a disinfectant-soaked paper towel. the cabinet with a disinfectant-soaked paper towel.

Discard contaminated disposable materials using Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI). (e.g., autoclave or BFI).

Place contaminated reusable items in biohazard bags, Place contaminated reusable items in biohazard bags, autoclavable pans with lids or wrap in newspaper autoclavable pans with lids or wrap in newspaper before autoclaving and cleanup. before autoclaving and cleanup.

Expose non-autoclavable materials to disinfectant (15 Expose non-autoclavable materials to disinfectant (15 minute contact time) before removal from the BSC.minute contact time) before removal from the BSC.


Remove protective clothing used during Remove protective clothing used during cleanup and place in a biohazard bag for cleanup and place in a biohazard bag for autoclaving. autoclaving.

Run cabinet 10 minutes after cleanup before Run cabinet 10 minutes after cleanup before resuming work or turning cabinet off.resuming work or turning cabinet off.

Biohazardous SpillsBiohazardous SpillsBiohazardous SpillsBiohazardous Spills Inside a CentrifugeInside a Centrifuge

Clear area of all personnel. Clear area of all personnel. Wait 30 minutes for aerosol to settle before attempting to Wait 30 minutes for aerosol to settle before attempting to

clean up spill. clean up spill. Wear a lab coat, safety glasses and gloves during cleanup. Wear a lab coat, safety glasses and gloves during cleanup. Remove rotors and buckets to nearest biological safety Remove rotors and buckets to nearest biological safety

cabinet for cleanup. cabinet for cleanup. Thoroughly disinfect inside of centrifuge. Thoroughly disinfect inside of centrifuge. Discard contaminated disposable materials using Discard contaminated disposable materials using

appropriate biohazardous waste disposal procedures appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI).(e.g., autoclave or BFI).


Biohazardous SpillsBiohazardous Spills


Requirements for the Bi-Annual InspectionRequirements for the Bi-Annual Inspection

Laboratory InspectionsLaboratory Inspections Every two yearsEvery two years Information NeededInformation Needed

PI’s name, department, building, room number, lab PI’s name, department, building, room number, lab manager, phone, mail code, email, etc. manager, phone, mail code, email, etc.

Laboratory Safety ManualLaboratory Safety Manual Protocols or Standard Operating ProceduresProtocols or Standard Operating Procedures Complete inventory of the Biohazardous materialsComplete inventory of the Biohazardous materials

• prions; genomic sequences;viroids; viruses; prions; genomic sequences;viroids; viruses; rickettsiae/chlamydia; bacteria;parasites; plants/plant, rickettsiae/chlamydia; bacteria;parasites; plants/plant, pathogens; animals; human/primate blood; human body pathogens; animals; human/primate blood; human body fluids, cells and tissuesfluids, cells and tissues

IBC approval forms and number(s) IBC approval forms and number(s) Emergency call-listEmergency call-list Outline of Annual Safety Training Session and Risk BriefingOutline of Annual Safety Training Session and Risk Briefing Acknowledgement of Annual Safety Training and Risk Acknowledgement of Annual Safety Training and Risk

BriefingBriefing

Laboratory InspectionsLaboratory Inspections Facility/EquipmentFacility/Equipment

Things we will expect and look for:Things we will expect and look for: Airflow from lower-hazard to higher-hazard Airflow from lower-hazard to higher-hazard

areas areas Designated clean area Designated clean area Any hazardous material in designated clean area Any hazardous material in designated clean area Neat or cluttered work areas Neat or cluttered work areas Biosafety cabinet: make, model, size, number Biosafety cabinet: make, model, size, number Negative-pressure thimble connection on Negative-pressure thimble connection on

biosafety cabinet biosafety cabinet Biosafety cabinet certificationBiosafety cabinet certification

• At least once per yearAt least once per year• Date of last certificationDate of last certification

Laboratory InspectionsLaboratory Inspections

Facility/EquipmentFacility/Equipment Things we will expect and look for:Things we will expect and look for:

Decontamination of biosafety cabinet Decontamination of biosafety cabinet before use before use

Decontamination of biosafety cabinet after Decontamination of biosafety cabinet after use use

• Neat or cluttered grate in biosafety cabinet Neat or cluttered grate in biosafety cabinet

• Neat or cluttered work area in biosafety cabinet Neat or cluttered work area in biosafety cabinet

• HEPA filter on vacuum line HEPA filter on vacuum line

• How full is the suction flaskHow full is the suction flask AutoclaveAutoclave

• Make, model, frequency of autoclave calibration, log Make, model, frequency of autoclave calibration, log

Laboratory InspectionsLaboratory Inspections Facility/EquipmentFacility/Equipment

Things we will expect and look for:Things we will expect and look for: CentrifugeCentrifuge

• make, model, condition of centrifuge bucket, make, model, condition of centrifuge bucket, condition of centrifuge rotors (check for condition of centrifuge rotors (check for stress cracks),stress cracks),

• condition of centrifuge interior (check for condition of centrifuge interior (check for residue buildup), log residue buildup), log

Spill-kit availabilitySpill-kit availability

Laboratory InspectionsLaboratory Inspections Work PracticesWork Practices

Things we will expect and look for:Things we will expect and look for: Aerosol-generating procedures and steps taken to Aerosol-generating procedures and steps taken to

control them control them Effective use of biosafety cabinets Effective use of biosafety cabinets Surface decontamination: disinfectant used, Surface decontamination: disinfectant used,

contact time, frequency contact time, frequency Are lab coats worn or not ?Are lab coats worn or not ? Are safety glasses required; if so, are they worn Are safety glasses required; if so, are they worn

or not ?or not ? Any evidence of eating in the lab areasAny evidence of eating in the lab areas

Interview of laboratory personnelInterview of laboratory personnel


Requirements for the Bi-annual inspectionRequirements for the Bi-annual inspection


Working with Biohazardous Materials: The role of Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential the PI in informing the laboratorian of potential hazards in the workplacehazards in the workplace

Working with Biohazardous Materials: The role of the PI in Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the informing the laboratorian of potential hazards in the workplaceworkplace

Working with Biohazardous Materials: The role of the PI in Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the informing the laboratorian of potential hazards in the workplaceworkplace

The role and responsibilities of the PI with The role and responsibilities of the PI with respect to Biosafety and Animalsrespect to Biosafety and Animals The Principal Investigator (PI) is responsible The Principal Investigator (PI) is responsible

for full compliance with the federal and state for full compliance with the federal and state regulations, NIH and CDC Guidelines, and regulations, NIH and CDC Guidelines, and institutional requirements for research institutional requirements for research involving biohazardous materials. involving biohazardous materials.

The PI is also responsible for ensuring that The PI is also responsible for ensuring that the reporting requirements are fulfilled and the reporting requirements are fulfilled and will be held accountable for any reporting will be held accountable for any reporting lapses.lapses.

The PI must:The PI must: Acquaint members of his/her laboratory Acquaint members of his/her laboratory

with risks associated with working with with risks associated with working with biohazardous materials and agentsbiohazardous materials and agents..

Develop a biosafety plan which identifies the Develop a biosafety plan which identifies the hazards present in his/her laboratoryhazards present in his/her laboratory

Identify specific practices and procedures Identify specific practices and procedures that need be followed in order to reduce the that need be followed in order to reduce the risk of working with these biohazardous risk of working with these biohazardous agents.agents.

Working with Biohazardous Materials: The role of the PI Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the in informing the laboratorian of potential hazards in the workplaceworkplace


The PI should be familiar with the current MUSC The PI should be familiar with the current MUSC safety policy entitled “Working with Biohazardous safety policy entitled “Working with Biohazardous Materials”Materials” It is accepted practice to give It is accepted practice to give specific trainingspecific training relevant to relevant to

requirements for working in an environment with requirements for working in an environment with biohazardous materials that may result in deleterious effects to biohazardous materials that may result in deleterious effects to laboratorians, any fetus they may be carrying, or close laboratorians, any fetus they may be carrying, or close associates such as household contacts. associates such as household contacts.

Laboratory personnel should be Laboratory personnel should be advised of specific hazardsadvised of specific hazards and and be required to read and to follow the biosafety plan prepared be required to read and to follow the biosafety plan prepared by the PI. by the PI.

Special care should be taken to advise at risk populations. Special care should be taken to advise at risk populations.

An at risk population includes, but is not limited to, An at risk population includes, but is not limited to, immunocompromised individuals, those individuals who are immunocompromised individuals, those individuals who are pregnant, and individuals of child bearing agepregnant, and individuals of child bearing age.



The PI should be familiar with current MUSC safety policy entitled The PI should be familiar with current MUSC safety policy entitled “Working with Biohazardous Materials”“Working with Biohazardous Materials”

Specific biohazards to pregnant women and their Specific biohazards to pregnant women and their fetuses include, but are not limited to, those agents in fetuses include, but are not limited to, those agents in the the TORCHTORCH group group

T, T, Toxoplasma gondiiToxoplasma gondii, O, , O, Treponema pallidumTreponema pallidum (syphilis), R, (syphilis), R, rubella, C, cytomegalovirus (CMV), and H, herpes simplex rubella, C, cytomegalovirus (CMV), and H, herpes simplex virus. virus.

However, there is also evidence that a number of other However, there is also evidence that a number of other viruses including, but not limited to, viruses including, but not limited to, adenovirusadenovirus, , coxsackiecoxsackie virusvirus, , Epstein-BarrEpstein-Barr virusvirus, hepatitis B virus, human , hepatitis B virus, human parvovirus, and varicella-zoster virus may result in adverse parvovirus, and varicella-zoster virus may result in adverse pregnancy outcomes. pregnancy outcomes.

Further, bacterial agents of special concern are those Further, bacterial agents of special concern are those classified as BSL3 agents and those BSL2 agents with known classified as BSL3 agents and those BSL2 agents with known consequences to the fetus such as consequences to the fetus such as Streptococcus agalactiae, Streptococcus agalactiae, group B Streptococcus (GBS) and group B Streptococcus (GBS) and ListeriaListeria..



The PI should be familiar with current MUSC safety The PI should be familiar with current MUSC safety policy entitled “Working with Biohazardous Materials”policy entitled “Working with Biohazardous Materials”

Role of the LaboratorianRole of the Laboratorian In addition to participating in training, reading In addition to participating in training, reading

and following the biosafety plan, it is also the and following the biosafety plan, it is also the responsibility of the laboratorian to inform their responsibility of the laboratorian to inform their immediate supervisor of any change in their immediate supervisor of any change in their health status (such as pregnancy, taking health status (such as pregnancy, taking medications resulting in reduced immunity etc.).medications resulting in reduced immunity etc.).

Furthermore, the laboratorian may wish to Furthermore, the laboratorian may wish to consult with student or employee health and/or consult with student or employee health and/or their personal physician to seek guidance with their personal physician to seek guidance with respect to how best to manage the risk.respect to how best to manage the risk.



The PI should be familiar with current MUSC safety The PI should be familiar with current MUSC safety policy entitled “Working with Biohazardous Materials”policy entitled “Working with Biohazardous Materials”

Management PlanManagement Plan

Appropriate action should be taken by the PI Appropriate action should be taken by the PI /supervisor to safeguard the health of the /supervisor to safeguard the health of the individual and, if necessary, the developing fetus. individual and, if necessary, the developing fetus.

A written, confidential, signed plan outlining the A written, confidential, signed plan outlining the management of the specific risk shall be placed in management of the specific risk shall be placed in the laboratorian’s personnel file acknowledging the laboratorian’s personnel file acknowledging their understanding and acceptance of the their understanding and acceptance of the management plan.management plan.



Management PlanManagement Plan




Management PlanManagement Plan Questions to Ask ?Questions to Ask ?

Hazard LevelsHazard Levels MSDS for MicrobesMSDS for Microbes




Management PlanManagement Plan Questions to Ask ?Questions to Ask ?

Hazard LevelsHazard Levels• MSDS for MicrobesMSDS for Microbes

– Issue AerosolIssue Aerosol– 150 pfu is 150 pfu is

infectious intra-infectious intra-nasallynasally

• IncubationIncubation– 1-10 days1-10 days

• CommunicabilityCommunicability– YesYes

http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/



Questions to Ask ?Questions to Ask ? Current LiteratureCurrent Literature



Am J. Obstet Gynecol 2003; Am J. Obstet Gynecol 2003; 189189(3): 758-763(3): 758-763

ApplicabilityApplicability Guidance and procedures associated with the policy Guidance and procedures associated with the policy

apply to all MUSC activities and its employees and apply to all MUSC activities and its employees and students who might be exposed to reproductive or students who might be exposed to reproductive or developmental hazards in the routine conduct of their developmental hazards in the routine conduct of their actions at MUSCactions at MUSC

Visitors who do not qualify as employees and/or Visitors who do not qualify as employees and/or students must involve their home institution in any students must involve their home institution in any discussion of accommodations. discussion of accommodations.

Contractors resident to campus must consult their Contractors resident to campus must consult their respective reproductive and developmental safety respective reproductive and developmental safety and health policies of their respective employers for and health policies of their respective employers for accommodations. accommodations.




Working with Biohazardous Materials: The role of Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential the PI in informing the laboratorian of potential hazards in the workplacehazards in the workplace

Biosafety at MUSCBiosafety at MUSC

Learning Objectives Units 1-7Learning Objectives Units 1-7 Understand the role and responsibilities of the PI with respect Understand the role and responsibilities of the PI with respect

to Biosafety and Animalsto Biosafety and Animals Understand the underlying principles of biosafety as they Understand the underlying principles of biosafety as they

relate to conducting safe and effective science on campusrelate to conducting safe and effective science on campus Be able to locate resources that will facilitate the determination Be able to locate resources that will facilitate the determination

of the relative biosafety risk associated with a planned of the relative biosafety risk associated with a planned experimentexperiment

Recognize the significance and need for conducting Recognize the significance and need for conducting experiments at different biosafety containment levelsexperiments at different biosafety containment levels

Understand the differences between BSL1 and BSL2Understand the differences between BSL1 and BSL2


Recognize the role each piece of Recognize the role each piece of Personal Protective Equipment Personal Protective Equipment plays plays in safeguarding the health and welfare of the laboratorian and in safeguarding the health and welfare of the laboratorian and communitycommunity

Be familiar with the proper use of Biological Safety Cabinets (BSC)Be familiar with the proper use of Biological Safety Cabinets (BSC) Recognize the inherent risk with using centrifuges in the laboratory Recognize the inherent risk with using centrifuges in the laboratory

with a special emphasis on risks associated with infectious agentswith a special emphasis on risks associated with infectious agents Understand the significance of good Laboratory HousekeepingUnderstand the significance of good Laboratory Housekeeping Be able to clean up a minor biological spill within a BSCBe able to clean up a minor biological spill within a BSC Understand what is required of you during the bi-annual Laboratory Understand what is required of you during the bi-annual Laboratory

Biological Safety Inspection Biological Safety Inspection Understand the signficance of working with biohazardous Understand the signficance of working with biohazardous

Materials specifically the role of the PI in informing the Materials specifically the role of the PI in informing the laboratorian of potential hazards in the workplace as well as laboratorian of potential hazards in the workplace as well as the personal responsibility required of each employee and the personal responsibility required of each employee and studentstudent

Questions ? Questions ? Questions ? Questions ?

IBC websiteIBC website

Date post:	30-Mar-2015
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Biosafety at MUSC IBC website IBC website Michael G. Schmidt, Ph.D., Chair, Biosafety Committee.

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