UNIVERSITY OF TEXAS MEDICAL BRANCH

Notification of Use

BSL3/4 and CDC/USDA Regulated Agents

The purpose of this document is to ensure adequate review of occupational safety and health precautions and the procedures for use, handling, storage, and disposal of biohazardous agents. As the Principal Investigator (P.I.) or Supervisor, you should be fully aware of the specifics of potential hazards associated with the agents used in your work area.

THIS MAY BECOME A PUBLIC DOCUMENT; DO NOT INSERT PROPRIETARY OR SECURITY SENSITIVE INFORMATION.

Type of Submission: New Renewal

Type of Agents: Biological Agent Recombinant Material

Select Agent: Yes No

The information provided in this document is accurate to the best of my knowledge. I am familiar with, the agree to abide by the provisions set forth in this plan as approved by the University of Texas Medical Branch Biological Safety and Institutional BioSafety Committees, the UTMB Safety Manual, the UTMB Institutional Handbook of Operating Procedures (IHOP), 42 CFR 73, 7 CFR 331, 9 CFR 121, the BioSafety in Medical and Biomedical Laboratories, NIH/CDC, most current edition, and the NIH guidelines for recombinant material.

When using recombinant DNA, I agree to comply with the NIH Guidelines for Research Involving Recombinant DNA Molecules.

I accept responsibility for training all laboratory workers involved in the research project described in this “Notification of Use” before commencing any work.

P.I. (Signature) Title Extension Date Submitted

Responsible for Research

P.I. (Printed Name) / Department / Route

BIOLOGICAL SAFETY COMMITTEE USE ONLY

DATE APPROVED ______DATE FOR RESUBMISSION ______

______

Chairman (Signature) Printed Name

SECTION I: General information

NOU are renewed every 5 years. Annual updates must be submitted (form sent by EHS annually). Amendments to the NOU do not change renewal date, original approval dates apply.

1.  List agent(s):

Attach a copy of the biological material safety data sheet if available,

(http://www.phac-aspc.gc.ca/msds-ftss/index.html).

2.  Goal of the project (1-2 sentences):

3.  Description of use (include techniques used for in-vitro, in-vivo and vector work-do not copy detailed protocols or grant information, this section should typically be about ½ page long):

4.  Location: Building: / Room(s):

5.  Can this agent infect humans? No Yes

If yes, is the infection associated with replication in humans or is it abortive (no infectious progeny; for example viral replicons or defective adenoviral vecotrs)?

Replication Abortive

If yes, can it cause disease in: - healthy humans? No Yes Unknown

- Immunocompromised people? No Yes Unknown

6.  Is immunization required for work at the listed biosafety level? No Yes

Is immunization recommended at the listed biosafety level? No Yes

(other than Yellow Fever vaccine for Keiller/GNL BSL3/ GNL BSL3E)

List vaccine:

If this is not an FDA-licensed vaccine (e.g. it is an IND product available through the U.S. Army Special Immunizations Program or a product available outside the U.S.), check here, consult the supplemental Vaccination Risk Assessment Form, and include vaccination information in your description of personnel experience and skill on question No. 12)

7.  Is medical surveillance recommended prior to commencement of work? No Yes

(Other than the respiratory surveillance and baseline serum program for BSL3/ BSL3E and Medical clearance for BSL4)

If yes, please explain (Post exposure management is considered part of occupational health exposure management)

8.  Have you educated your staff regarding safe handling and decontamination procedures for the agents in this NOU? No Yes

9.  Risk Assessment:

a.  Describe pathogenicity, including disease incidence and severity.

b.  Describe potential routes of laboratory transmission

c.  Describe agent stability (environmental stability in the lab, susceptibility to decontamination)

d.  What is the infectious dose?

e.  What is the concentration (number of infectious organism per unit volume) and the volume of the material being handled at one time? (maximum volume and concentration cultured at one time)

f.  What is the origin of the infectious material (may refer to geographic location, host or nature of source), from where will you receive your agent?

g.  Summarize any data available from animal studies (pathogenicity, infectivity and route of transmission in animal)?

h.  Is there an effective prophylaxis or therapeutic intervention available? Specify prophylaxis or therapeutic intervention?

10.  Evaluation of Dual Use potential- experiments of concern (National Research Council Biotechnology Research in an Age of Terrorism, or “Fink Committee Report”). If answer “yes”, please explain in detail, use additional sheets as needed.

i.  Would this research demonstrate how to render a vaccine (if applicable) ineffective?

No Yes Specify:

j.  Would this research confer resistance to therapeutically useful antibiotics and antiviral agents?

No Yes Specify:

k.  Would this research enhance the virulence of a pathogen, or render a non-pathogen virulent?

No Yes Specify:

l.  Would this research increase transmissibility of this pathogen?

No Yes Specify:

m.  Would this research alter the host range of this pathogen?

No Yes Specify:

n.  Would this research enable the evasion of diagnostic/detection modalities of this agent?

No Yes Specify:

o.  Would this research enable the weaponization of a biological agent or toxin? (the term “weaponization” includes, for example, enhancing the aerosol delivery of pathogens, new techniques for microencapsulation, and the synthesis of viral pathogens.)

No Yes Specify:

11.  What systems (cells and bacteria) are you using to propagate or study the agent(s) listed?

12.  At what Biosafety Level will this agent be used?

BSL2 BSL3 BSL3 E BSL4

13.  Check the protective clothing or equipment used when handling this agent:

Lab coat/ gloves/ eye protection (BSL2) Cover gown/ gloves/ eye protection (BSL3)

Scrubs, cover gown/booties/gloves/ Dover/ Delta suits (BSL4)

eye protection (BSL3E)

PAPR (Racal/Max Air) N95 respirator

Full/half face respirator Face Shield

Safety Centrifuge/blender Other specify:

Chemical Fume Hood Room location:

Biological Safety Cabinet Room location:

14.  Method for disposal of biohazardous waste:

Placed in red bag for disposal.

Autoclaved, then placed in regular trash.

Chemically disinfect, then placed in regular trash.

Chemical disinfection of bulk liquid, then poured down sanitary drain (BSL3) or effluent treatment system (BSL3E and BSL4).

Autoclaved, then packaged for incineration. (BSL 3 & 4 only)

15.  List disinfectant(s) used for surface decontamination and spills:

Cavicide Phenolic germicidal 70% alcohol

MicroChem Other specify: 10% Bleach

16.  Experience and skill level of personnel working with the material. Include experience with agents/cells, techniques used, animal species/procedures and biosafety levels. If applicable, please indicate if each staff member will be working with animals and/or arthropods and their specific experience with each.

If you are using rDNA/RNA, please fill out section II N/A

If you are using a select agent, please fill out section III N/A

If you are planning any animal work, please fill out section IV N/A

If you are planning any arthropod vector work, please fill out section V N/A

1

BSL3-4-Revised 03/12

SECTION II: Recombinant DNA / RNA

http://oba.od.nih.gov/rdna/nih_guidelines_oba.html

Definition of a recombinant molecule per NIH-OBA guidelines

In the context of the NIH Guidelines, recombinant DNA molecules are defined as either: (i) molecules that are constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell, or (ii) molecules that result from the replication of those described in (i) above.

Synthetic DNA segments which are likely to yield a potentially harmful polynucleotide or polypeptide (e.g., a toxin or a pharmacologically active agent) are considered as equivalent to their natural DNA counterpart. If the synthetic DNA segment is not expressed in vivo as a biologically active polynucleotide or polypeptide product, it is exempt from the NIH Guidelines.

Genomic DNA of plants and bacteria that have acquired a transposable element, even if the latter was donated from a recombinant vector no longer present, are not subject to the NIH Guidelines unless the transposon itself contains recombinant DNA.

1.  Goal of the project (1-2 sentences):

2.  Description of the project (specific to recombinant work):

3.  Location: Building: / Room(s):

4.  At what biosafety level will you be conducting the cloning portion (not rescue) of your research:

BSL1 BSL2 BSL4 (full length viruses)

5.  Does this project involve synthetic DNA? Yes No

6.  Is this vector/insert commercially purchased? Yes No

(No clone manipulation will be done in your lab.)

Please provide the company’s name and product number:

7.  Is this vector/insert provided by a collaborator? Yes No

(No clone manipulation will be done in your lab.)

Please provide the collaborator’s institution:

8.  Please fill out the following table

Source of Insert DNA/RNA
(Species) / Name of Vector(s) Used
List them for all steps in the cloning process / Name of Host(s) Used (bacteria or virus or cells)
List them for all steps in the cloning process / Nature of Inserted DNA Sequences / If an attempt will be made to obtain expression of a foreign gene, indicate the protein that will be produced
9.  Answer the following questions, use additional pages if needed
Additional info needed / specify
a) Does the inserted gene encode a known oncogene? / Yes
No
b) Does the viral DNA integrate into the host genome? / Yes
No
c) Does the modification have the potential to increase the replication capacity of virus?
Does the modification increase the pathogenicity of the agent?
Does the modification change the host range of the agent? / Yes
No / See section I10: Evaluation of Dual use
e) Does the inserted gene have the potential for altering the cell cycle? / Yes
No
h) Is there a probability of generating replication-competent viruses? / Yes
No
Will this work involve the deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally? Other than E. coli cloning selection
Could this acquisition compromise the use of the drug to control disease agents in humans, veterinary medicine, or agriculture? / Yes
No
Yes
No / A-1a
Will this experiment involve the cloning of toxin molecules with LD50 of less than 100 Nanograms per Kilogram body weight (e.g., microbial toxins such as the botulinum toxins, tetanus toxin, diphtheria toxin, and Shigella dysenteriae neurotoxin)? / Yes No / B-1
a) Provide LD50 ng/Kg of body weight:
b) Is this molecule lethal to vertebrates at 100 ng to 100 mg per kg body weight? / Yes
No
c) Is the cloning done in Escherichia coli K-12? / Yes
No
Will this experiment involving the deliberate transfer of recombinant DNA, or DNA/RNA derived from recombinant DNA, into one or more human research participants. Contact EHS before submitting / Yes
No / C-1
Will this experiment involve using Risk Group 2, Risk Group 3, Risk Group 4, or Restricted Agents as Host-Vector Systems? If no please go to section D-2. / Yes
No / D-1
Will this experiment involve the introduction of recombinant DNA into Risk Group 2 agents? / Yes
No / D-1-a
Will this experiment involve the introduction of recombinant DNA into Risk Group 3 agents? / Yes
No / D-1-b
Will this experiment involve the introduction of recombinant DNA into Risk Group 4 agents? / Yes
No / D-1-C
Will this experiment involve the introduction of recombinant DNA into restricted agents? / Yes
No / D-1-d
Will this experiment involve DNA From Risk Group 2, Risk Group 3, Risk Group 4, or Restricted Agents to be Cloned into Nonpathogenic Prokaryotic or Lower Eukaryotic Host-Vector Systems? If no please go to section D-3 / Yes
No / D-2
a) Please list prokaryotes used
b) Please list lower eukaryotes used
Will this experiment involve DNA from Risk Group 2 or Risk Group 3 agents to be transferred into nonpathogenic prokaryotes or lower eukaryotes? / Yes
No / D-2-a
Is the DNA from Risk Group 4 agents present in a given recombinant is totally and irreversibly defective fraction of the agent's genome? / Yes
No / if no: work must be done at BSL4
Will this experiment involve DNA from restricted agents to be transferred into nonpathogenic prokaryotes or lower eukaryotes? / Yes
No / D-2-b
Will this experiment involve the Use of Infectious DNA/RNA Viruses in Tissue Culture Systems?
Will this experiment involve the Use of Defective DNA/RNA Viruses in the Presence of Helper Virus in Tissue Culture Systems? If no please go to section D-4 / Yes
No / D-3
Will this experiment involve the use of infectious or defective Risk Group 2 viruses in the presence of helper virus? / Yes
No / D-3-a
Will this experiment involve the use of infectious or defective Risk Group 3 viruses in the presence of helper virus? / Yes
No / D-3-b
Will this experiment involve the use of infectious or defective Risk Group 4 viruses in the presence of helper virus? / Yes
No / D-3-c
Will this experiment involve the use of infectious or defective restricted poxviruses in the presence of helper virus? Contact EHS before submission. / Yes
No / D-3-d
Will this experiment involve the use of infectious or defective viruses in the presence of helper virus, which are not covered in Sections III-D-3-a through III-D-3-d? / Yes
No / D-3-e
Will this experiment involve the use of Whole Animals? This section covers experiments involving whole animals in which the animal's genome has been altered by stable introduction of recombinant DNA, or DNA derived therefrom, into the germ-line (transgenic animals) and experiments involving viable recombinant DNA-modified microorganisms tested on whole animals.
If no please go to section E. / Yes
No / D-4
Will this experiment involve any of the following:
1) Recombinant DNA, or DNA or RNA molecules derived therefrom, from any source except for greater than two-thirds of eukaryotic viral genome may be transferred to any non-human vertebrate or any invertebrate organism.
2) Animals that contain sequences from viral vectors, which do not lead to transmissible infection either directly or indirectly as a result of complementation or recombination in animals.
3) For experiments involving recombinant DNA-modified Risk Groups 2, 3, 4, or restricted organisms. It is important that the investigator demonstrate that the fraction of the viral genome being utilized does not lead to productive infection. / Yes
No / D-4-a
Will this experiment involve the use of replicating recombinant risk group 2-3-4 and restricted agents involving whole animals, including transgenic animals, and not covered by Sections III-D-1 or III-D-4-a? / Yes