Download entire manual. This section has been reviewed and updated as needed: May 2015.


  1. Foreword

Chapter 1: Introduction

  1. Introduction
  2. The Occurrence Of Laboratory-Associated Infections
  3. Evolution Of National Biosafety Guidelines
  4. Risk Criteria For Establishing Ascending Levels Of Containment
  5. Agent Summary Statements
  6. Biosecurity
  7. Using BMBL
  8. Looking Ahead

Chapter 2: Biological Risk Assessment

  1. Biological Risk Assessment
  2. Hazardous Characteristics Of An Agent
  3. Hazardous Characteristics Of Laboratory Procedures
  4. Potential Hazards Associated With Work Practices, Safety Equipment And Facility Safeguards
  5. An Approach To Assess Risks And Select Appropriate Safeguards
  6. Conclusion

Chapter 3: Principles Of Biosafety

  1. Principles Of Biosafety
  2. Laboratory Practice And Technique
  3. Safety Equipment (Primary Barriers And Personal Protective Equipment)
  4. Facility Design And Construction (Secondary Barriers)
  5. Biosafety Levels
  6. Animal Facilities
  7. Clinical Laboratories
  8. Importation And Interstate Shipment Of Certain Biomedical Materials
  9. Select Agents

Chapter 4: Laboratory Biosafety Level Criteria

  1. Laboratory Biosafety Level Criteria
  2. Biosafety Level 1
  3. Biosafety Level 2
  4. Biosafety Level 3
  5. Biosafety Level 4
  6. Table 1: Summary Of Recommended Biosafety Levels For Infectious Agents
  7. Section I: Designating Biological Safety Level 2 at UNC
    1. Forms, Documents, and Training
    2. Laboratory Facilities
    3. Safety Equipment
    4. Special Practices
  8. Appendix A: Biological Safety Level 2 Checklist
  9. Appendix B: Cryogenic Preservation of Biological Materials

Chapter 5: Vertebrate Animal Biosafety Level Criteria for Vivarium Research Facilities

  1. Vertebrate Animal Biosafety Level Criteria for Vivarium Research Facilities
  2. Animal Biosafety Level 1
  3. Animal Biosafety Level 2
  4. Animal Biosafety Level 3
  5. Animal Biosafety Level 4
  6. Table 1: Summary of Recommended Biosafety Levels for Infectious Agents

Chapter 6: Principles of Laboratory Biosecurity

  1. Principles of Laboratory Biosecurity
  2. Biosafety and Biosecurity
  3. Risk Management Methodology
  4. Elements of a Biosecurity Program

Chapter 7: Occupation Health and Immunoprophylaxis

  1. Occupational Health and Immunoprophylaxis
  2. Occupational Health Support Service Elements
  3. Vaccines
  4. Occupational Health in the BSL-4 Setting

Chapter 8: Agent Summary Statements

  1. Section I: Bacterial Agents
    1. Agent: Bacillus anthracis
    2. Agent: Bordetella pertussis
    3. Agent: Brucella species
    4. Agent: Burkholderia mallei
    5. Agent: Burkholderia pseudomallei
    6. Agent: Campylobacter (C. jejuni subsp. jejuni, C. coli, C. fetus subsp. fetus, C. upsaliensis)
    7. Agent: Chlamydia psittaci (Chlamydophila psittaci), C. trachomatis, C. pneumoniae (Chlamydophila pneumoniae)
    8. Agent: Neurotoxin Producing Clostridia species
    9. Agent: Clostridium tetani and Tetanus toxin
    10. Agent: Corynebacterium diphtheria
    11. Agent: Francisella tolarensis
    12. Agent: Helicobacter species
    13. Agent: Legionella pneumophila and Other Legionella-Like Agents
    14. Agent: Leptospira
    15. Agent: Listeria monocytogenes
    16. Agent: Mycobacterium leprae
    17. Agent: Mycobacterium tubercolosis complex
    18. Agent: Mycobacterium spp. Other Than M. tubercolosiscomplex and M. leprae
    19. Agent: Neisseria gonorrhoeae
    20. Agent: Neisseria meningitides
    21. Agent: Salmonella serotypes, Other Than S. typhi
    22. Agent: Salmonella typhi
    23. Agent: Shiga toxin (Verocytotoxin)-ProducingEscherichia coli
    24. Agent: Shigella
    25. Agent: Treponema pallidum
    26. Agent: Vibrio enteritis species (V. cholerae, V. parahaemolyticus)
    27. Agent: Yersinia pestis
    28. Section Ia: Policy For Laboratory Personnel Who Are Working With Neisseria meningitidis
  2. Section II: Fungal Agents
    1. Agent: Blastomyces dermatitidis
    2. Agent: Coccidioides immitis and Coccidioides posadasii
    3. Agent: Cryptococcus neoformans
    4. Agent: Histoplasma capsolatum
    5. Agent: Sporothrix schenckii
    6. Agent: Dermatophytes (Epidermophyton, Microsporum, And Trichophyton)
    7. Agent: Miscellaneous Molds
  3. Section III: Parasitic Agents
    1. General Issues
    2. Agent: Blood And Tissue Protozoal Parasites
    3. Agent: Intestinal Protozoal Parasites
    4. Agent: Trematode Parasites
    5. Agent: Cestode Parasites
    6. Agent: Nematode Parasites
  4. Section IV: Rickettsial Agents
    1. Agent: Coxiella burnetii
    2. Agents: Rickettsia prowazekii, Rickettsia typhi (R. mooseri), Orientia (Rickettsia) Tsutsugamushi and Spotted Fever Group Agents of Human Disease;Rickettsia rickettsii, Rickettsia conorii, Rickettsia akari, Rickettsia australis, Rickettsia siberica, and Rickettsia japonicum
  5. Section V: Viral Agents
    1. Agent: Hantaviruses
    2. Agent: Hendra Virus (Formerly Known as Equine Morbillivirus) and Nipah Virus
    3. Agent: Hepatitis A Virus, Hepatitis E Virus
    4. Agent: Hepatitis B Virus, Hepatitis C Virus (Formerly Known as Nona Nonb Virus), Hepatitis D Virus
    5. Agent: Herpesvirus simiae (Cerocopithecine herpesvirusI, Herpes B Virus)
    6. Agent: Human Herpes Virus
    7. Agent: Influenza
    8. Agent: Lymphocytic Choriomeningitis Virus
    9. Agent: Poliovirus
    10. Agent: Poxviruses
    11. Agent: Rabies Virus (and Related Lyssaviruses)
    12. Agent: Retroviruses, Including Human and Simian Immunodeficiency Viruses (HIV and SIV)
    13. Agent: Severe Acute Respiratory Syndrome (SARS) Coronavirus
    14. Section Va: Reassortant Influenza A Viruses
  6. Section VI: Arboviruses and Related Zoonotic Viruses
    1. Arboviruses and Related Zoonotic Viruses
    2. Viruses With BSL-2 Containment Recommended
    3. Viruses With BSL-3 Containment Recommended
    4. Viruses With BSL-4 Containment Recommendations
    5. Dealing With Unknown Arboviruses
    6. Agent: West Nile Virus (WNV)
    7. Agent: Eastern Equine Encephalitis (EEE) Virus, Venezuelan Equine Encephalitis (VEE) Virus, And Western Equine Encephalitis (WEE) Virus
    8. Agent: Rift Valley Fever Virus (RVFV)
    9. Table 3: Agent: Alphabetic Listing of 597 Arboviruses and Hemorrhagic Fever Viruses
  7. Section VII: Toxin Agents
    1. Agent: Botolinum Neurotoxin
    2. Agent: Staphylococcal enterotoxins (SE)
    3. Agent: Ricin Toxin
    4. Agent: Selected Low Molecolar Weight (LMW) Toxins
  8. Section VIII: Prion Diseases
    1. Prion Diseases
    2. Occupational Infections
    3. Natural Modes of Infection
    4. Laboratory Safety
    5. Special Issues
  9. Section IX: Viral Vectors
    1. Viral Vectors
    2. Adenovirus Vectors
    3. Adeno-Associated Virus (AAV) Vectors
    4. Epstein-Barr Virus Vectors
    5. Herpesvirus Vectors
    6. Retroviruses: Murine Leukemia Virus (MLV) Vectors
    7. Retroviruses: Lentivirus Vectors
    8. Poxviruses Vectors
    9. Replication Competency Testing

Chapter 9: Primary Containment for Biohazards: Selection, Installation and Use of Biological Safety Cabinets

  1. Introduction
  2. The High Efficiency Particolate Air (HEPA) Filter and the Development of Biological Containment Devices
  3. Biological Safety Cabinets
  4. Other Laboratory Hazards and Risk Assessment
  5. BSC Use by the Investigator: Work Practices and Procedures
  6. Facility and Engineering Requirements
  7. Certification Of BSCS
  8. Policy on the Use of Flammable Gases in Biological Safety Cabinets
  9. Section I: Policy on the Use of Flammable Gases in Biological Safety Cabinets

Chapter 10: Decontamination and Disinfection

  1. Decontamination And Disinfection
  2. Environmentally Mediated Infection Transmission
  3. Principles of Sterilization and Disinfection
  4. Decontamination in the Microbiology Laboratory
  5. Special Infectious Agent Issues
  6. Section I: Biological Waste Disposal Policy

Chapter 11: Transportation of Infectious Substances

  1. Transportation of Infectious Substances

Chapter 12: Agricultural Pathogen Biosafety

The contents of this chapter were provided by USDA. All questions regarding its contents should be forwarded to the USDA.

  1. Introduction
  2. BSL-3-Ag
  3. BSL-3, Enhanced
  4. Pathogens of Veterinary Significance
  5. Summaries of Selected Agriculture Agents
  6. Additional Information

Chapter 13: Arthropod Containment Guidelines (ACG)

  1. Arthropod Containment Guidelines (ACG)

Chapter 14: Select Agents and Toxins

  1. Select Agents and Toxins
  2. Dual Use Research of Concern Policy

Chapter 15: Integrated Pest Management (IPM)

  1. Integrated Pest Management (IPM)

Chapter 16: Working With Human, NHP and Other Mammalian Cells and Tissues

  1. Working With Human, NHP and Other Mammalian Cells and Tissues
  2. Section I: Bloodborne Pathogen Exposure Control Plan for Laboratories
    1. Regulation
    2. Definitions
    3. Exposure Determination
    4. Universal Precaution
    5. Epidemiology
    6. Facilities and Practices
    7. Engineering Controls
    8. Work Practice Controls
    9. Personal Protective Equipment
    10. Housekeeping
    11. Waste Disposal
    12. HIV and HBV Research Laboratories
    13. Hepatitis B Vaccination
    14. Post-Exposure Procedures
    15. Post-Exposure Evaluation and Follow-Up
    16. Training
    17. Inspection
    18. Exposure Control For UNC-CH Employees Located in Other Counties
    19. Appendix A: Job Classifications
    20. Appendix B: Hepatitits B Declination Form
  3. Safety Practices for Work with Non-Human Primate Tissues and Body Fluids

Chapter 17: Guidelines for Work With Toxins of Biological Origin

  1. Guidelines for Work With Toxins of Biological Origin

Chapter 18: NIH Oversight of Research Involving Recombinant Biosafety Issues

  1. NIH Oversight of Research Involving Recombinant Biosafety Issues
  2. Section I: Animal Experiments Covered Under the NIH Guidelines for Research Involving Recombinant DNA Molecoles
  3. Section II: Transgenic Animals and the Use of Recombinant DNA in Animals FAQs
  4. Section III: Incident Reporting

Chapter 19: Resources for Information

  1. Resources for Information


  1. Acronyms