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

2. Biosafety at MUSC Learning Objectives Learning Objectives 1. Understand the role and responsibilities of the PI with respect to Biosafety and Animals 2. Understand the underlying principles of biosafety as they relate to conducting safe and effective science requiring the use of biohazardous materials on campus 3. Be able to locate resources that will facilitate the determination of the relative biosafety risk associated with a planned experiment 4. Recognize the significance and need for conducting experiments at different biosafety containment levels 5. Understand the differences between BSL1 and BSL2

3. Biosafety at MUSC Learning Objectives continued: Learning Objectives continued: 6. Recognize the role each piece of Personal Protective Equipment plays in safeguarding the health and welfare of the laboratorian and community 7. Be familiar with the proper use of Biological Safety Cabinets (BSC) 8. Recognize the inherent risk with using centrifuges in the laboratory with a special emphasis on risks associated with infectious agents 9. Understand the significance of good laboratory housekeeping 10. Be able to clean up a minor biological spill within a BSC 11. Understand what is required of you during the bi- annual Laboratory Biological Safety Inspection

4. Biosafety at MUSC Learning Objectives continued: Learning Objectives continued: 12. Understand the signficance of working with biohazardous Materials specifically the role of the PI in informing the laboratorian of potential hazards in the workplace as well as the personal responsibility required of each employee and student

5. Biosafety at MUSC Unit 1 Unit 1 The role and responsibilities of the PI with respect to Biosafety and Animals at MUSC The role and responsibilities of the PI with respect to Biosafety and Animals at MUSC

6. Biosafety in Biomedical Laboratories at MUSC The role and responsibilities of the PI with respect to Biosafety and Animals The role and responsibilities of the PI with respect to Biosafety and Animals A full-time faculty or professional staff member with an advanced degree must agree to accept responsibility for the project and serve as the PI. A full-time faculty or professional staff member with an advanced degree must agree to accept responsibility for the project and serve as the PI. Student researchers, residents, and postdoctoral fellows cannot be identified as principal investigators. Student researchers, residents, and postdoctoral fellows cannot be identified as principal investigators. The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. The PI is also responsible for ensuring that the reporting requirements are fulfilled and will be held accountable for any reporting lapses. The PI is also responsible for ensuring that the reporting requirements are fulfilled and will be held accountable for any reporting lapses.

7. DefinitionDefinition Biohazard Biohazard An agent of biological origin that has the capacity to produce deleterious effects on humans, animals, plants and insects. These include microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from insects, animals and plants. An agent of biological origin that has the capacity to produce deleterious effects on humans, animals, plants and insects. These include microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from insects, animals and plants.

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

9. Biosafety in Microbiological and Biomedical Laboratories The role and responsibilities of the PI with respect to Biosafety and Animals The role and responsibilities of the PI with respect to Biosafety and Animals The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm PDF http://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm

10. Biosafety at MUSC Questions for Unit 1 Questions for Unit 1 The role and responsibilities of the PI with respect to Biosafety and Animals at MUSC The role and responsibilities of the PI with respect to Biosafety and Animals at MUSC

11. Biosafety at MUSC Unit 2 Unit 2 Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous materials at MUSC Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous materials at MUSC

12. DefinitionDefinition Biohazard Biohazard An agent of biological origin that has the capacity to produce deleterious effects on humans, animals, plants and insects. These include microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from insects, animals and plants. An agent of biological origin that has the capacity to produce deleterious effects on humans, animals, plants and insects. These include microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from insects, animals and plants.

13. Biosafety in Microbiological and Biomedical Laboratories Principles of Biosafety Principles of Biosafety Containment Containment The purpose of containment is to reduce or eliminate exposure of: The purpose of containment is to reduce or eliminate exposure of: laboratory workers,laboratory workers, other personsother persons the environment the environment Safe methods for managing infectious agents in the laboratory Safe methods for managing infectious agents in the laboratory Environment Environment The location where the agent being handled, maintained or stored. The location where the agent being handled, maintained or stored.

14. Infectious Agents and the lab worker Modes of Infection Modes of Infection Infections preceded by overt personal accidents, which include: Infections preceded by overt personal accidents, which include: Inoculation Inoculation resulting from pricking, jabbing or cutting the skin with contaminated instruments such as:resulting from pricking, jabbing or cutting the skin with contaminated instruments such as: – hypodermic needles, scalpels and glassware; – from 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 eyes Splashing into the face and eyes Spillage and direct contact Spillage and direct contact

15. Infectious Agents and the lab worker Infections not preceded by personal accidents: Infections not preceded by personal accidents: Aerosols, droplets and fomites. Aerosols, droplets and fomites. These are speculated (from Pikes 1976 data) to be responsible for up to 82% percent of all laboratory-acquired infections. These are speculated (from Pikes 1976 data) to be responsible for up to 82% percent of all laboratory-acquired infections. Aerosols are a cloud of very small liquid droplets produced whenever energy is applied to a liquid, and such liquid is allowed to escape into the environment. Aerosols are a cloud of very small liquid droplets produced whenever energy is applied to a liquid, and such liquid is allowed to escape into the environment. Centrifugation is a good source (plasmid preps)Centrifugation is a good source (plasmid preps)

16. Infectious Agents and the lab worker There are many regulations in place to forestall the problem of laboratory-acquired infections. There are many regulations in place to forestall the problem of laboratory-acquired infections. Responsibility for compliance with the regulations Responsibility for compliance with the regulations Primarily to the Principal Investigator Primarily to the Principal Investigator Secondarily, with the laboratory staff. Secondarily, with the laboratory staff. In general a large number of organisms that would ordinarily be innocuous can be infective in immune compromised persons. In general a large number of organisms that would ordinarily be innocuous can be infective in immune compromised persons. Therefore, additional and more stringent measures must be established by the PI in an effort to prevent the occurrence of lab-acquired infections in such individuals. Therefore, additional and more stringent measures must be established by the PI in an effort to prevent the occurrence of lab-acquired infections in such individuals.

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

18. Biosafety in Microbiological and Biomedical Laboratories Primary containment Primary containment Protection from exposure to infectious agents, is provided by both good microbiological technique and the use of appropriate safety equipment. Protection from exposure to infectious agents, is provided by both good microbiological technique and the use of appropriate safety equipment. Secondary containment Secondary containment Protection of the environment external to the laboratory from exposure to infectious materials Protection of the environment external to the laboratory from exposure to infectious materials Facility design Facility design Operational practices Operational practices

19. Biosafety in Microbiological and Biomedical Laboratories Three elements of containment include: Three elements of containment include: 1. Laboratory practice and technique, 2. Safety equipment Bunsen burners vs. Bactincinerators Bunsen burners vs. Bactincinerators Cages and other barriers Cages and other barriers 3. Facility design The risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements. The risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements.

20. Biosafety in Microbiological and Biomedical Laboratories Laboratory practice and technique Laboratory practice and technique Standard microbiological practices and techniques Standard microbiological practices and techniques Persons working with infectious agents or potentially infected materials must be aware of potential hazards, and must be trained and proficient in the practices and techniques required for handling such material safely. Persons working with infectious agents or potentially infected materials must be aware of potential hazards, and must be trained and proficient in the practices and techniques required for handling such material safely. The director or person in charge of the laboratory is responsible for providing or arranging for appropriate training of personnel. The director or person in charge of the laboratory is responsible for providing or arranging for appropriate training of personnel.

21. Biosafety in Microbiological and Biomedical Laboratories A scientist trained and knowledgeable in appropriate laboratory techniques, safety procedures, and hazards associated with handling infectious agents must direct laboratory activities. A scientist trained and knowledgeable in appropriate laboratory techniques, safety procedures, and hazards associated with handling infectious agents must direct laboratory activities. Each laboratory should develop or adopt a biosafety or operations manual which identifies the hazards that will or may be encountered, and which specifies practices and procedures designed to minimize or eliminate risks. Each laboratory should develop or adopt a biosafety or operations manual which identifies the hazards that will or may be encountered, and which specifies practices and procedures designed to minimize or eliminate risks. All personnel should be advised of special hazards and should be required to read and to follow the required practices and procedures. All personnel should be advised of special hazards and should be required to read and to follow the required practices and procedures. Each individual in the laboratory should acknowledge that they have been advised and briefed as to the laboratorys plan and to the risks that they will be exposed. Each individual in the laboratory should acknowledge that they have been advised and briefed as to the laboratorys plan and to the risks that they will be exposed. There is no such thing as zero risk! There is no such thing as zero risk!

22. Biosafety at MUSC Questions for Unit 2 Questions for Unit 2 Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous materials at MUSC Principles of Biosafety as they relate to the safe conduct of science requiring the use of biohazardous materials at MUSC

23. Biosafety at MUSC Unit 3 Unit 3 How to determine the relative biosafety risk associated with a planned experiment How to determine the relative biosafety risk associated with a planned experiment

24. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for Microbes MSDS for Microbes MSDS for Microbes MSDS for Microbes Standard Microbiological Practices Standard Microbiological Practices Special Practices Special Practices Containment Equipment Containment Equipment http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

25. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for Microbes MSDS for Microbes MSDS for Microbes MSDS for Microbes http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

26. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for Microbes MSDS for Microbes MSDS for Microbes MSDS for Microbes Issue AerosolIssue Aerosol –150 pfu is infectious intra- nasally IncubationIncubation –1-10 days CommunicabilityCommunicability –Yes http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

27. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels Disinfectants Disinfectants BleachBleach Physical Inactivation Physical Inactivation Survival outside host Survival outside host Type 3 survival 10 days on paperType 3 survival 10 days on paper Type 2 survived 3-8 weeks on environmental surfaces at room temperature.Type 2 survived 3-8 weeks on environmental surfaces at room temperature. http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

28. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels 10 case of lab infections 10 case of lab infections Special Hazards Special Hazards Contact with feces from infected animals Contact with feces from infected animals Spills Spills Disposal Disposal http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

29. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for Microbes MSDS for Microbes MSDS for Microbes MSDS for Microbes Standard Microbiological Practices Standard Microbiological Practices Special Practices Special Practices Containment Equipment Containment Equipment http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/

30. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Infectious Agents Infectious Agents Work may only be conducted with prior approval of the IBC regardless of the safety classification of the agent Work may only be conducted with prior approval of the IBC regardless of the safety classification of the agent You must follow the requirements as specified in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories Manual You must follow the requirements as specified in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories Manual Biosafety in Microbiological and Biomedical Laboratories Manual Biosafety in Microbiological and Biomedical Laboratories Manual Containment requirements may be subject to modification by the IBC Containment requirements may be subject to modification by the IBC

31. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Define the risk Define the risk Infectious Agents List Infectious Agents List Experimental Protocol Experimental Protocol How big ? How big ? Aerosols ? Aerosols ? Animals ? Animals ?

32. Laboratory Containment Levels for Biological Research Involving Potential Biohazards SECTION II - Principles of Biosafety SECTION II - Principles of BiosafetyPrinciples of BiosafetyPrinciples of Biosafety Recommended Biosafety Levels for Infectious Agents in the Laboratory Recommended Biosafety Levels for Infectious Agents in the Laboratory Summary of Recommended Biosafety Levels for Activities in Which Experimentally or Naturally Infected Vertebrate Animals Are Used Summary of Recommended Biosafety Levels for Activities in Which Experimentally or Naturally Infected Vertebrate Animals Are Used http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s2.htm http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm PDF

33. Laboratory Containment Levels for Biological Research Involving Potential Biohazards Section III Laboratory- Biosafety Level Criteria Section III Laboratory- Biosafety Level Criteria Section III Laboratory- Biosafety Level Criteria Section III Laboratory- Biosafety Level Criteria 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 Criteria Section IV-Vertebrate Animal Biosafety Level Criteria Section IV-Vertebrate Animal Biosafety Level Criteria Section IV-Vertebrate Animal Biosafety Level Criteria ABSL 1, ABSL 1, ABSL 2, ABSL 2, ABSL 3, ABSL 3, ABSL 4 ABSL 4 http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s3.htm http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s4.htm

34. Laboratory Containment Levels for Biological Research Involving Potential Biohazards SECTION V - Risk Assessment SECTION V - Risk Assessment SECTION V SECTION V SECTION VI- Recommended Biosafety Levels For Infectious Agents and Infected Animals SECTION VI- Recommended Biosafety Levels For Infectious Agents and Infected Animals SECTION VI- SECTION VI- Section VII- Agent Summary Statements Section VII- Agent Summary Statements Section VII- Section VII- http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s5.htm http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s6.htm http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4s7.htm

35. Recombinant DNA, Gene Therapy and Transgenics Guidelines for Research Involving Recombinant DNA Molecules Guidelines for Research Involving Recombinant DNA Molecules Or in PDF at this site Or in PDF at this site Scope Scope Safety Safety Experiments Governed Experiments Governed Roles and Responsibilities Roles and Responsibilities http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Gdlnes_lnk_2002z.pdf

36. Recombinant DNA, Gene Therapy and Transgenics Guidelines for Research Involving Recombinant DNA Molecules Guidelines for Research Involving Recombinant DNA Molecules Classification of Human Etiologic Agents on the Basis of Hazard Classification of Human Etiologic Agents on the Basis of Hazard Appendix B-I. Risk Group 1 (RG1) Agents Appendix B-I. Risk Group 1 (RG1) Agents Appendix B-II. Risk Group 2 (RG2) Agents Appendix B-II. Risk Group 2 (RG2) Agents Appendix B-III. Risk Group 3 (RG3) Agents Appendix B-III. Risk Group 3 (RG3) Agents Appendix B-IV. Risk Group 4 (RG4) Agents Appendix B-IV. Risk Group 4 (RG4) Agents

37. Recombinant DNA, Gene Therapy and Transgenics Guidelines for Research Involving Recombinant DNA Molecules Guidelines for Research Involving Recombinant DNA Molecules Physical and Biological Containment for Recombinant DNA Research Involving Animals Physical and Biological Containment for Recombinant DNA Research Involving Animals Appendix Q-I. General Considerations Appendix 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 Levels Appendix 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 Q Appendix Q-III. Footnotes and References for Appendix Q http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_02.htm

38. Recombinant DNA, Gene Therapy and Transgenics Guidelines for Research Involving Recombinant DNA Molecules Guidelines for Research Involving Recombinant DNA Molecules Whats Exempt? Whats 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 viruses Those that are not in organisms or viruses Those that consist entirely of DNA segments from a single non-chromosomal or viral DNA source, though one or more of the segments may be synthetic Those that consist entirely of DNA segments from a single non-chromosomal or viral DNA source, though one or more of the segments may be synthetic Those that consist entirely of DNA from a prokaryotic host including its 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 well established physiological means. Those that consist entirely of DNA from a prokaryotic host including its 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 well established physiological means. Those that consist entirely of DNA from a eukaryotic host including its indigenous chloroplasts, mitochondria or plasmids (excluding viruses) when propagated in that host. Those that consist entirely of DNA from a eukaryotic host including its indigenous chloroplasts, mitochondria or plasmids (excluding viruses) when propagated in that host. Bottom line… PCR and gel running.. Bottom line… PCR and gel running..

39. Biosafety at MUSC Questions Unit 3 Questions Unit 3 How to determine the relative biosafety risk associated with a planned experiment How to determine the relative biosafety risk associated with a planned experiment

40. Biosafety at MUSC Unit 4 Unit 4 Specific Requirements for Biosafety Levels 1 to 3 Specific Requirements for Biosafety Levels 1 to 3 PPE PPE Biological Safety Cabinets (BSCs) Biological Safety Cabinets (BSCs) Risks associated with centrifugation and proper protocols Risks associated with centrifugation and proper protocols

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

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

65. Human Tissue and Cell Culture Human blood, blood products, body fluids and tissues are listed as potentially Hazardous Biological Materials Human blood, blood products, body fluids and tissues are listed as potentially Hazardous Biological Materials Biosafety Level 2 Practices and procedures MUST be followed when handling Biosafety Level 2 Practices and procedures MUST be followed when handling BloodBlood Blood ProductsBlood Products Body fluidsBody fluids TissuesTissues Under no circumstance shall anyone work with cells derived from themselves or from first degree relatives since the host immune systems may not provide adequate protectionUnder no circumstance shall anyone work with cells derived from themselves or from first degree relatives since the host immune systems may not provide adequate protection See Green book for greater detailSee Green book for greater detail

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

67. Human Tissue and Cell Culture Cell Culture Cell Culture The following cell lines are at Biosafety Level 2 or higher The following cell lines are at Biosafety Level 2 or higher 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 packaging systems All cell lines exposed to or transformed by amphotropic packaging systems All clinical material (e.g., samples of human tissues and fluids obtained after surgical resection or autopsy) All clinical material (e.g., samples of human tissues and fluids obtained after surgical resection or autopsy) All cell lines new to the laboratory (until proven to be free of all adventitious agents) All cell lines new to the laboratory (until proven to be free of all adventitious agents) All mycoplasma-containing cell lines All mycoplasma-containing cell lines

68. Personal Protective Equipment PPE is used to protect you from contact with biohazardous materials PPE is used to protect you from contact with biohazardous materials Graphic credit: http://www-ehs.ucsd.edu/bio11.htm

69. Personal Protective Equipment If the science you are conducting requires you to wear a N95 Respirator (Mycobacterium tuberculosis) Occupational Health and Safety must FIT-TEST you prior to you starting work! If the science you are conducting requires you to wear a N95 Respirator (Mycobacterium tuberculosis) Occupational Health and Safety must FIT-TEST you prior to you starting work!

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

84. Laboratory Housekeeping Good housekeeping is essential for Good housekeeping is essential for Reducing risks Reducing risks Protecting the integrity of biological experiments Protecting the integrity of biological experiments Keep the laboratory neat and free of clutter. Keep the laboratory neat and free of clutter.

85. Biosafety at MUSC Questions Unit 4 Questions Unit 4 Specific Requirements for Biosafety Levels 1 to 3 Specific Requirements for Biosafety Levels 1 to 3 PPE PPE Biological Safety Cabinets (BSCs) Biological Safety Cabinets (BSCs) Risks associated with centrifugation and proper protocols Risks associated with centrifugation and proper protocols Cell Culture Cell Culture Spills Spills

86. Biosafety at MUSC Unit 5 Unit 5 Biohazardous Spills Biohazardous Spills

87. Biohazardous Spills Outside the Biosafety cabinet Outside the Biosafety cabinet Clear area of all personnel. Clear area of all personnel. Wait at least 15 minutes for aerosol to settle before entering spill area. Wait at least 15 minutes for aerosol to settle before entering spill area. Remove any contaminated clothing and place in biohazard bag to be autoclaved. Remove any contaminated clothing and place in biohazard bag to be autoclaved. Put on a disposable gown, safety glasses and gloves. Put on a disposable gown, safety glasses and gloves.

88. Biohazardous Spills Outside the Biosafety cabinet Outside the Biosafety cabinet Initiate cleanup with disinfectant as follows: Initiate cleanup with disinfectant as follows: 1.Place dry paper towel on spill (to absorb liquids); then layer a second set of disinfectant soaked paper towels over the spill. 2.Encircle the spill with additional disinfectant being careful to minimize aerosolization while assuring adequate contact. 3.Decontaminate all items within spill the area. 4.Allow minimum of 15 minutes contact time to ensure germicidal action of disinfectant. 5.Wipe equipment with appropriate disinfectant. 6.Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI). 7.Disinfect reusable items.

89. Biohazardous Spills Inside the Biosafety Cabinet (BSC) Inside the Biosafety Cabinet (BSC) Wait at least five minutes to allow the BSC to contain aerosols. Wait at least five minutes to allow the BSC to contain 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 contact time. Apply disinfectant and allow a minimum of 15 minutes contact time. Wipe up spillage with disposable disinfectant-soaked paper towel. Wipe up spillage with disposable disinfectant-soaked paper towel. Place used cleanup items in the appropriate biohazard waste container Place used cleanup items in the appropriate biohazard waste container

90. Biohazardous Spills Inside the Biosafety Cabinet (BSC) Inside the Biosafety Cabinet (BSC) Wipe the walls, work surface and any equipment in the cabinet with a disinfectant-soaked paper towel. Wipe the walls, work surface and any equipment in the cabinet with a disinfectant-soaked paper towel. Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI). Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI). Place contaminated reusable items in biohazard bags, autoclavable pans with lids or wrap in newspaper before autoclaving and cleanup. Place contaminated reusable items in biohazard bags, autoclavable pans with lids or wrap in newspaper before autoclaving and cleanup. Expose non-autoclavable materials to disinfectant (15 minute contact time) before removal from the BSC. Expose non-autoclavable materials to disinfectant (15 minute contact time) before removal from the BSC.

91. Biohazardous Spills Inside the Biosafety Cabinet (BSC) Inside the Biosafety Cabinet (BSC) Remove protective clothing used during cleanup and place in a biohazard bag for autoclaving. Remove protective clothing used during cleanup and place in a biohazard bag for autoclaving. Run cabinet 10 minutes after cleanup before resuming work or turning cabinet off. Run cabinet 10 minutes after cleanup before resuming work or turning cabinet off.

92. Biohazardous Spills Inside a Centrifuge Inside a Centrifuge Clear area of all personnel. Clear area of all personnel. Wait 30 minutes for aerosol to settle before attempting to clean up spill. Wait 30 minutes for aerosol to settle before attempting to 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 cabinet for cleanup. Remove rotors and buckets to nearest biological safety cabinet for cleanup. Thoroughly disinfect inside of centrifuge. Thoroughly disinfect inside of centrifuge. Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI). Discard contaminated disposable materials using appropriate biohazardous waste disposal procedures (e.g., autoclave or BFI).

93. Biosafety at MUSC Questions Unit 5 Questions Unit 5 Biohazardous Spills Biohazardous Spills

94. Biosafety at MUSC Unit 6 Unit 6 Requirements for the Bi-Annual Inspection Requirements for the Bi-Annual Inspection

95. Laboratory Inspections Every two years Every two years Information Needed Information Needed PIs name, department, building, room number, lab manager, phone, mail code, email, etc. PIs name, department, building, room number, lab manager, phone, mail code, email, etc. Laboratory Safety Manual Laboratory Safety Manual Protocols or Standard Operating Procedures Protocols or Standard Operating Procedures Complete inventory of the Biohazardous materials Complete inventory of the Biohazardous materials prions; genomic sequences;viroids; viruses; rickettsiae/chlamydia; bacteria;parasites; plants/plant, pathogens; animals; human/primate blood; human body fluids, cells and tissues prions; genomic sequences;viroids; viruses; rickettsiae/chlamydia; bacteria;parasites; plants/plant, pathogens; animals; human/primate blood; human body fluids, cells and tissues IBC approval forms and number(s) IBC approval forms and number(s) Emergency call-list Emergency call-list Outline of Annual Safety Training Session and Risk Briefing Outline of Annual Safety Training Session and Risk Briefing Acknowledgement of Annual Safety Training and Risk Briefing Acknowledgement of Annual Safety Training and Risk Briefing

96. Laboratory Inspections Facility/Equipment Facility/Equipment Things we will expect and look for: Things we will expect and look for: Airflow from lower-hazard to higher-hazard areas Airflow from lower-hazard to higher-hazard 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 biosafety cabinet Negative-pressure thimble connection on biosafety cabinet Biosafety cabinet certification Biosafety cabinet certification At least once per yearAt least once per year Date of last certification Date of last certification

97. Laboratory Inspections Facility/Equipment Facility/Equipment Things we will expect and look for: Things we will expect and look for: Decontamination of biosafety cabinet before use Decontamination of biosafety cabinet before use Decontamination of biosafety cabinet after use Decontamination of biosafety cabinet after use Neat or cluttered grate in biosafety cabinetNeat or cluttered grate in biosafety cabinet Neat or cluttered work area in biosafety cabinetNeat or cluttered work area in biosafety cabinet HEPA filter on vacuum lineHEPA filter on vacuum line How full is the suction flaskHow full is the suction flask Autoclave Autoclave Make, model, frequency of autoclave calibration, logMake, model, frequency of autoclave calibration, log

98. Laboratory Inspections Facility/Equipment Facility/Equipment Things we will expect and look for: Things we will expect and look for: Centrifuge Centrifuge make, model, condition of centrifuge bucket, condition of centrifuge rotors (check for stress cracks),make, model, condition of centrifuge bucket, condition of centrifuge rotors (check for stress cracks), condition of centrifuge interior (check for residue buildup), logcondition of centrifuge interior (check for residue buildup), log Spill-kit availability Spill-kit availability

99. Laboratory Inspections Work Practices Work Practices Things we will expect and look for: Things we will expect and look for: Aerosol-generating procedures and steps taken to control them Aerosol-generating procedures and steps taken to control them Effective use of biosafety cabinets Effective use of biosafety cabinets Surface decontamination: disinfectant used, contact time, frequency Surface decontamination: disinfectant used, contact time, frequency Are lab coats worn or not ? Are lab coats worn or not ? Are safety glasses required; if so, are they worn or not ? Are safety glasses required; if so, are they worn or not ? Any evidence of eating in the lab areas Any evidence of eating in the lab areas Interview of laboratory personnel Interview of laboratory personnel

101. Biosafety at MUSC Questions Unit 6 Questions Unit 6 Requirements for the Bi-annual inspection Requirements for the Bi-annual inspection

102. Biosafety at MUSC Unit 7 Unit 7 Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

103. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace The role and responsibilities of the PI with respect to Biosafety and Animals The role and responsibilities of the PI with respect to Biosafety and Animals The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. The Principal Investigator (PI) is responsible for full compliance with the federal and state regulations, NIH and CDC Guidelines, and institutional requirements for research involving biohazardous materials. The PI is also responsible for ensuring that the reporting requirements are fulfilled and will be held accountable for any reporting lapses. The PI is also responsible for ensuring that the reporting requirements are fulfilled and will be held accountable for any reporting lapses.

104. The PI must: The PI must: Acquaint members of his/her laboratory with risks associated with working with biohazardous materials and agents. Acquaint members of his/her laboratory with risks associated with working with biohazardous materials and agents. Develop a biosafety plan which identifies the hazards present in his/her laboratory Develop a biosafety plan which identifies the hazards present in his/her laboratory Identify specific practices and procedures that need be followed in order to reduce the risk of working with these biohazardous agents. Identify specific practices and procedures that need be followed in order to reduce the risk of working with these biohazardous agents. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

105. The PI should be familiar with the current MUSC safety policy entitled Working with Biohazardous Materials The PI should be familiar with the current MUSC safety policy entitled Working with Biohazardous Materials It is accepted practice to give specific training relevant to requirements for working in an environment with biohazardous materials that may result in deleterious effects to laboratorians, any fetus they may be carrying, or close associates such as household contacts. It is accepted practice to give specific training relevant to requirements for working in an environment with biohazardous materials that may result in deleterious effects to laboratorians, any fetus they may be carrying, or close associates such as household contacts. Laboratory personnel should be advised of specific hazards and be required to read and to follow the biosafety plan prepared by the PI. Laboratory personnel should be advised of specific hazards and be required to read and to follow the biosafety plan prepared 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, immunocompromised individuals, those individuals who are pregnant, and individuals of child bearing age An at risk population includes, but is not limited to, immunocompromised individuals, those individuals who are pregnant, and individuals of child bearing age. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

106. The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials Specific biohazards to pregnant women and their fetuses include, but are not limited to, those agents in the TORCH group Specific biohazards to pregnant women and their fetuses include, but are not limited to, those agents in the TORCH group T, Toxoplasma gondii, O, Treponema pallidum (syphilis), R, rubella, C, cytomegalovirus (CMV), and H, herpes simplex virus. T, Toxoplasma gondii, O, Treponema pallidum (syphilis), R, rubella, C, cytomegalovirus (CMV), and H, herpes simplex virus. However, there is also evidence that a number of other viruses including, but not limited to, adenovirus, coxsackie virus, Epstein-Barr virus, hepatitis B virus, human parvovirus, and varicella-zoster virus may result in adverse pregnancy outcomes. However, there is also evidence that a number of other viruses including, but not limited to, adenovirus, coxsackie virus, Epstein-Barr virus, hepatitis B virus, human parvovirus, and varicella-zoster virus may result in adverse pregnancy outcomes. Further, bacterial agents of special concern are those classified as BSL3 agents and those BSL2 agents with known consequences to the fetus such as Streptococcus agalactiae, group B Streptococcus (GBS) and Listeria. Further, bacterial agents of special concern are those classified as BSL3 agents and those BSL2 agents with known consequences to the fetus such as Streptococcus agalactiae, group B Streptococcus (GBS) and Listeria. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

107. The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials Role of the Laboratorian Role of the Laboratorian In addition to participating in training, reading and following the biosafety plan, it is also the responsibility of the laboratorian to inform their immediate supervisor of any change in their health status (such as pregnancy, taking medications resulting in reduced immunity etc.). In addition to participating in training, reading and following the biosafety plan, it is also the responsibility of the laboratorian to inform their immediate supervisor of any change in their health status (such as pregnancy, taking medications resulting in reduced immunity etc.). Furthermore, the laboratorian may wish to consult with student or employee health and/or their personal physician to seek guidance with respect to how best to manage the risk. Furthermore, the laboratorian may wish to consult with student or employee health and/or their personal physician to seek guidance with respect to how best to manage the risk. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

108. The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials The PI should be familiar with current MUSC safety policy entitled Working with Biohazardous Materials Management Plan Management Plan Appropriate action should be taken by the PI /supervisor to safeguard the health of the individual and, if necessary, the developing fetus. Appropriate action should be taken by the PI /supervisor to safeguard the health of the individual and, if necessary, the developing fetus. A written, confidential, signed plan outlining the management of the specific risk shall be placed in the laboratorians personnel file acknowledging their understanding and acceptance of the management plan. A written, confidential, signed plan outlining the management of the specific risk shall be placed in the laboratorians personnel file acknowledging their understanding and acceptance of the management plan. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

109. Management Plan Management Plan http://www.cdc.gov/od/ohs/pdffiles/ Module%202%20-%20Biosafety.pdf Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

110. Management Plan Management Plan Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for Microbes MSDS for Microbes MSDS for Microbes MSDS for Microbes http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/ Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

111. Management Plan Management Plan Questions to Ask ? Questions to Ask ? Hazard Levels Hazard Levels MSDS for MicrobesMSDS for MicrobesMSDS for MicrobesMSDS for Microbes –Issue Aerosol –150 pfu is infectious intra- nasally IncubationIncubation –1-10 days CommunicabilityCommunicability –Yes http://www.hc-sc.gc.ca/pphb-dgspsp/msds-ftss/ Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

112. Questions to Ask ? Questions to Ask ? Current Literature Current Literature Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace Am J. Obstet Gynecol 2003; 189(3): 758-763

113. Applicability Applicability Guidance and procedures associated with the policy apply to all MUSC activities and its employees and students who might be exposed to reproductive or developmental hazards in the routine conduct of their actions at MUSC Guidance and procedures associated with the policy apply to all MUSC activities and its employees and students who might be exposed to reproductive or developmental hazards in the routine conduct of their actions at MUSC Visitors who do not qualify as employees and/or students must involve their home institution in any discussion of accommodations. Visitors who do not qualify as employees and/or students must involve their home institution in any discussion of accommodations. Contractors resident to campus must consult their respective reproductive and developmental safety and health policies of their respective employers for accommodations. Contractors resident to campus must consult their respective reproductive and developmental safety and health policies of their respective employers for accommodations. Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

114. Biosafety at MUSC Questions for Unit 7 Questions for Unit 7 Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace Working with Biohazardous Materials: The role of the PI in informing the laboratorian of potential hazards in the workplace

115. Biosafety at MUSC Learning Objectives Units 1-7 Learning Objectives Units 1-7 Understand the role and responsibilities of the PI with respect to Biosafety and Animals Understand the role and responsibilities of the PI with respect to Biosafety and Animals Understand the underlying principles of biosafety as they relate to conducting safe and effective science on campus Understand the underlying principles of biosafety as they relate to conducting safe and effective science on campus Be able to locate resources that will facilitate the determination of the relative biosafety risk associated with a planned experiment Be able to locate resources that will facilitate the determination of the relative biosafety risk associated with a planned experiment Recognize the significance and need for conducting experiments at different biosafety containment levels Recognize the significance and need for conducting experiments at different biosafety containment levels Understand the differences between BSL1 and BSL2 Understand the differences between BSL1 and BSL2

116. Biosafety at MUSC Learning Objectives continued: Learning Objectives continued: Recognize the role each piece of Personal Protective Equipment plays in safeguarding the health and welfare of the laboratorian and community Recognize the role each piece of Personal Protective Equipment plays in safeguarding the health and welfare of the laboratorian and community 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 with a special emphasis on risks associated with infectious agents Recognize the inherent risk with using centrifuges in the laboratory with a special emphasis on risks associated with infectious agents Understand the significance of good Laboratory Housekeeping Understand the significance of good Laboratory Housekeeping Be able to clean up a minor biological spill within a BSC Be able to clean up a minor biological spill within a BSC Understand what is required of you during the bi-annual Laboratory Biological Safety Inspection Understand what is required of you during the bi-annual Laboratory Biological Safety Inspection Understand the signficance of working with biohazardous Materials specifically the role of the PI in informing the laboratorian of potential hazards in the workplace as well as the personal responsibility required of each employee and student Understand the signficance of working with biohazardous Materials specifically the role of the PI in informing the laboratorian of potential hazards in the workplace as well as the personal responsibility required of each employee and student

117. Questions ? IBC website IBC website