Biosafety @ EH&S

Useful Resources

• Institutional Biosafety Plan (IBP)
• The Institutional Biosafety Plan (IBP) provides guidance on requirements for the safe use of regulated biological materials in research and teaching at UCLA. The IBP applies to all UCLA faculty, staff, students, contract employees, and other personnel working at locations where UCLA has management of the following biological materials:
   

  - Infectious agents (bacteria, viruses, fungi, parasites and prions) that can cause disease in healthy humans and/or significant environmental or agricultural impacts

  - Human and nonhuman primate materials (all fluids, tissues, excretions, secretions, primary cells or cell lines)

  - Select agents and select toxins (see Guidelines Regulating Select Agents & Toxins below)

  - Recombinant and synthetic nucleic acid molecules (see Guidelines Regulating Recombinant or Synthetic Nucleic Acid Molecules below)

  - Genetically modified animals and whole plants

• Biological Use Authorization (BUA) and Institutional Biosafety Committee (IBC) Approval
• A BUA is required for all research and teaching activities involving biological materials described in the IBP. BUAs must be submitted using SafetyNet and approved by the Institutional Biosafety Committee (IBC). More information can be found in the Institutional Biosafety Plan: Chapter 2.
  
  Go to SafetyNet
• Biosafety Levels (BSL) 
• All work practices and procedures must comply with the BSL containment standards outlined in the table below. More information on Animal Biosafety Levels (ABSLs) and Plant Biosafety Levels can be found in the Institutional Biosafety Plan: Chapter 5. Additional guidance is provided upon request.
  
  Summary of Recommended Biosafety Levels for Infectious Agents

  BSL	Agents	Required Safety Equipment	Facilities Requirements
  1	Not known to consistently cause disease in healthy human adults	PPE: laboratory coats and gloves; eye protection appropriate to hazards	Doors for access control Sink for hand washing Designed for cleaning (no cloth, rugs, carpets) Sturdy, chemical-resistant furniture Fly screens if windows open
  2	Associated with human disease, but lower risk of exposure Typical hazards relate to percutaneous injury, ingestion, mucous membrane exposure	BSL1 equipment plus   **BSC or other containment devices when aerosols/splashes are anticipated or high concentrations/volumes are used **Aerosol-tight centrifuge rotors or safety cups Respiratory protection in rooms with infected animals Protection of vacuum lines	BSL1 facility plus Doors must lock and be self-closing BSCs must be installed properly in facilities Eyewash station Inward flow of air without recirculation (new construction)
  2+		BSL2 equipment plus BSC required for all manipulation Additional PPE: Clothing with solid front (disposable gown recommended), surgical mask, double gloves	BSL2 facility
  3	Indigenous or exotic agents for aerosol transmission, low infectious dose Disease may have serious or lethal consequences or higher exposure risks	BSL2+ equipment plus Respiratory protection often more appropriate	BSL2+ facility plus Self-closing, double-door access Monitored and alarmed constant negative airflow, no reversal 100% exhaust, may need HEPA filtration Sealed seams and penetrations to permit gaseous decontamination Hands-free sinks Windows are sealed Physical separation from access corridors
  4	Dangerous and exotic agents that pose high risk of life-threatening disease, aerosol transmission. Emerging infectious diseases — NOT PERMITTED AT UCLA
  	**Additional considerations added to BSL1 for BSL1 enhanced (BSL1+)

• Biosafety Site Visits

• Biosafety Review

  A comprehensive review of the facilities, equipment and practices covered under each BUA is required when a new facility is set up and every 3 years thereafter or whenever significant changes are made that affect the risk assessment (e.g., containment level change, new major equipment or operations). During the review, EH&S Biosafety works with the PI or a designated representative to evaluate the efficacy of the biosafety program and to ensure that facility standards are maintained, engineering controls are utilized effectively, and risk mitigation plans documented in SOPs are implemented and meeting the needs of personnel. Following reviews, EH&S Biosafety will generate reports to summarize the findings. A follow-up visit will be scheduled within 30 days to address any outstanding issues. The IBC may choose to investigate repeat findings as outlined in the Policy on Investigating Allegations of Noncompliance.

  Schedule an appointment with EH&S Biosafety
• Guidelines Regulating Recombinant or Synthetic Nucleic Acid (r/sNA) Molecules

• Recombinant or synthetic nucleic acid (r/sNA) molecules are constructed outside of living cells. They are made by joining DNA or RNA segments (natural or synthetic) to DNA or RNA molecules that can replicate within a living cell. 
  
  It is mandatory that all UCLA researchers working with r/sNA molecules follow the National Institute of Health (NIH) Guidelines on Research Involving Recombinant DNA Molecules. 

  Experiments that Require NIH Director Approval and IBC Approval (III-A)	Minimum BSL	NIH Section
  Transfer of a drug resistance trait to microorganisms compromising ability to control disease agents in humans, veterinary medicine or agriculture.	Varies	III-A-1-a
  Experiments that Require NIH/OSP and IBC Approval (III-B)	Minimum BSL	NIH Section
  Cloning of toxin molecules with LD50 of less than 100 nanograms per kilogram body weight	Varies	III-B-1
  Experiments that Require IBC and IRB Approvals before Research Participant Enrollment (III-C)	Minimum BSL	NIH Section
  Transfer human gene into human research participant(s) using recombinant or synthetic nucleic acid molecules or DNA or RNA	Varies	III-C-1
  Experiments that Require IBC Approval (III-D)	Minimum BSL	NIH Section
  Adding nucleic acids to agents to Risk Group 2*	2	III-D-1-a
  Adding nucleic acids to agents to Risk Group 3*	3	III-D-1-b
  Adding nucleic acids to agents to Risk Group 4*	4	III-D-1-c
  Adding nucleic acids to restricted agents	4	III-D-1-d
  Adding nucleic acids from agents into nonpathogenic prokaryotes or lower eukaryotes	BSL1 or higher	III-D-2
  Nucleic acids in viral vectors Risk Group 2*	2	III-D-3-a
  Nucleic acids in viral vectors Risk Group 3*	3	III-D-3-b
  Nucleic acids in viral vectors Risk Group 4*	4	III-D-3-c
  Nucleic acids in viral vectors, such as restricted poxviruses	Varies	III-D-3-d
  Creation, breeding, transfer and experimentation with transgenic and knockout animals	Varies	Varies
  Nucleic acid-modified microorganisms tested on whole animals	BSL1 or higher	III-D-4
  Generation of transgenic plants or testing of nucleic acid-modified microorganisms or insects on whole plants at BSL2P+ or higher	BSL2P+ or higher	III-D-5
  Large-scale experiments greater than 10 liters of culture	BSL1 or higher	III-D-6
  Influenza virus experimentation: H2N2 (1957-1968)	3+	III-D-7
  Influenza virus experiments: Highly Pathogenic Avian (H5N1)	BSL2+ or higher	III-D-7-b
  Influenza virus experiments: 1918 H1N11	BSL3	III-D-7-c
  Experiments that Require IBC Approval (III-E)	Minimum BSL	NIH Section
  Use of viral vectors (<2/3 viral genome) in tissue culture	BSL1 or 2	III-E-1
  Generation of transgenic plants or testing of nucleic acid-modified microorganisms or insects on whole plants at BSL1P+ or BSL2P	BSL1P or 2P	III-E-2
  Generation of transgenic rodents (BSL1 only)	1	III-E-3
  Exempt Experiments (III-F)	Minimum BSL	NIH Section
  Synthetic nucleic acids that cannot replicate nor integrate on a living cell (i.e. oligos)	Varies	III-F-1
  Nucleic acids that are not in organisms, cells or viruses	Varies	III-F-2
  Exact nucleic acid sequence from a single source that exists contemporaneously in nature	Varies	III-F-3
  Prokaryotic host plasmids or viruses used only in that host or closely related strain	Varies	III-F-4
  Eukaryotic host nucleic acids used only in that host or closely related strain	Varies	III-F-5
  Nucleic acids entirely of DNA segments from different species that exchange DNA by known physiological processes	Varies	III-F-6
  Genomic DNA molecules that have acquired a transposable element	Varies	III-F-7
  Nucleic acids that do not present a significant risk to health or the environment (See NIH Guidelines Appendix C, Exemptions under Section III-F-8 for other classes of experiments which are exempt from the NIH Guidelines.)	Varies	III-F-8

  *More information on risk groups can be found at PHE Risk Group and the ABSA Risk Group Database. 
• Guidelines Regulating Escherichia coli Strain (E. coli)
• The the National Institute of Health (NIH) Guidelines on Research Involving Recombinant DNA Molecules states that with some exceptions, experimental use of Escherichia coli (E. coli) K-12 strain and its derivatives are exempt from the requirements of the NIH Guidelines.  This exempt status allows for faster review and approval of the Biohazard Use Authorization (BUA). 
  
  Below are lists of commonly used strains of E. coli and the NIH Guidelines category applicable to those strains.
   

  Ancestral E. coli The following E. coli laboratory strains are K-12 and therefore most research utilizing these strains is exempt from the NIH Guidelines (Section III-F, F-8, Appendix C-II).  Strain Designation Origin or Collection 58 Stanford Strain 679 Stanford Strain WG1 Wisconsin Strain E. coli K-12 Derivatives The following E. coli laboratory strains are K-12 derivatives and therefore most research utilizing these strains is exempt from the NIH Guidelines (Section III-F, F-8, Appendix C-II).  Strain Designation Origin or Collection Strain Designation Origin or Collection 5K Lab strain JC9387 Lab strain 58 Lab strain JM83 Lab strain 58-161 Lab strain JM101 Lab strain AB284 Lab strain KP7600 Lab strain AB311 Lab strain LE392 Lab strain AG1 Lab strain M15 Lab strain C600 Lab strain MB408 Lab strain Cavilli Hfr Lab strain MG1655 Lab strain DH1 Lab strain Novablue Novagen Dh5-alpha Lab strain P678 Lab strain DP50 Lab strain PA 309 Lab strain EMG2 Lab strain REG-12 Lab strain EPI100-T1R Lab strain S17-1 Lab strain H1443 Lab strain SCS-110 Strategen HB1001 Lab strain SM10 Lab strain Hfr3000 Lab strain STBL2 Invitrogen Hfr3000 X74 Lab strain STBL3 Invitrogen HMS 174 Novegen SURE Lab strain JM109 Lab strain TB1 NEB TG1 Lab strain WA704 Lab strain TOP10 Invitrogen W1485 Lab strain W1485 Lab strain W3110 Lab strain W208 Lab strain XL1-Blue Strategen W3110 Lab strain XL10-Gold Strategen W945 Lab strain XLOLR Strategen WA704 Lab strain Y10 Lab strain WG1 Lab strain YM2980 Lab strain E. coli NOT Derived from K-12 The following E. coli laboratory strains are not derived from K-12 and therefore any research utilizing these strains are not exempt from the NIH Guidelines (Section III-E).  Strain Designation Origin or Collection Strain Designation Origin or Collection B Lab strain K5808 Lab strain B-3 Lab strain Mach 1 Invitrogen B/R Lab strain Nissile 1917 Lab strain BL21 Novagen Rosetta Novagen BL23 NEB REG-118 Lab strain C Lab strain TOPP Strategen C41 Sigma Aldrich W Lab strain C43 Sigma Aldrich 25922 Lab strain FDA strain Seattle 1946 Lab strain     Pathogenic E. coli strains  The following E. coli strains are not exempt  from the NIH Guidelines (Section III-D). Strain Designation E. coli, all strains bearing K1 antigen E. coli enteroaggregative strain (EAEC) E. coli enterohaemorrhagic strain (EHEC) E. coli enteroinvesavie strain (EIEC) E. coli enteropathogenic strain (EPEC) E. coli enterotoxigenic strain (ETEC) E. coli Shiga toxin producing E. coli (O157:H7)

• Guidelines Regulating Select Agents & Toxins
• Select agent and toxin regulations require that any entity, facility and personnel involved in possession, use, or transfer of select agents and toxins register with the Federal Select Agent Program (FSAP). More information on can be found in the Institutional Biosafety Plan: Chapter 3.
   

  Tier 1 Select Agents and Toxins	Tier 1 can pose as a severe threat to public health and safety, and require additional training, containment, occupational health and security requirements (see 42 CFR 73 and 9 CFR 121).
  Permissible Amounts of Select Toxins	The UCLA IBC requires registration of permissible amounts of select toxins before acquisition of any amount to ensure proper biosafety and biosecurity measures are in place. This does not require registration with FSAP. Related documents & forms: Select Toxin Inventory Log Select Toxin Worksheet Select Toxin Due Diligence Form
  Dual Use Research of Concern	Dual use research of concern (DURC) refers to life sciences research involving 15 designated agents and toxins and 7 categories of experiments. These agents and experiments pose a significant threat with broad potential consequences to public health and safety, plants, animals, the environment, military materials and equipment, or national security.  At UCLA, the Dual Use Review Entity (DURE) identifies DURC and implements risk mitigation measures per UCLA Policy 995.

• Frequently Asked Questions (FAQs)
• How do I report an incident?
  If this an emergency, call 9-1-1. To reach the EH&S Hotline, call (310) 825-9797.
  
  How do I request a new biohazard door card?
  Complete the the Biohazard Door Card Request form. These door cards are for sub-rooms. If the door is public facing, complete the Universal Door Card Request form. 
  
  How often should my Biosafety Cabinet (BSC) be certified and how do I get it certified?
  BSCs should be certified annually. A TSS sticker on the BSC will note when it was last certified and the retest date. If your BSC needs to be serviced, please contact the phone number on the sticker.
  
  If you have a fume hood that you would like to get serviced, email fumehoods@ehs.ucla.edu to schedule a technician visit. 
  
  What should I use to disinfect?
  Please see the Decontamination SOP.
  
  I don't know who to contact. What do I do?
  Our EH&S Virtual Office Hours portal will allow you to schedule a time to speak with an EH&S representative. In the portal, click on the "i" for more information on each service EH&S provides. 
  
  Is there information on how to manage core facilities?
  Please see the Core Facility Management SOP. 
  
  What if I can new personnel?
  New personnel can use the Site Safety Orientation Checklist. 
  
  What training do I take if I will perform BSL-3 experiments?
  The BSL-3 training program is an extended mentorship and competency testing plan, which is conducted over several weeks in one of the UCLA BSL-3 facilities. Currently, the biggest prerequisite is to have an approved project which will take place in a UCLA BSL-3 lab at some point in the coming months. You can have whoever is coordinating your lab's BSL-3 experiment get in touch with the UCLA High Containment Director about scheduling your training. Please visit the High Containment website for more information. 


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