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UNIVERSITY OF TEXAS MEDICAL BRANCH
Notification of Use
for
Biological Agents and rDNA/ RNA
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 or 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 Agent
Biological Agent
Recombinant Material
The information provided in this document is accurate to the best of my knowledge. I am familiar with,
and agree to abide by the provisions set forth in this plan as approved by the University of Texas
Medical Branch Institutional Biosafety Committee, the UTMB Safety Manual, the UTMB Institutional
Handbook of Operating Procedures (IHOP), the CDC/NIH Biosafety in Microbiological and Biomedical
laboratories, most current edition, and NIH guidelines on recombinant work.
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 work.
P.I. (Signature)
Responsible for Research
Title
P.I. (Printed Name)
Extension
Department
Date Submitted
Route
BIOLOGICAL SAFETY COMMITTEE USE ONLY
DATE APPROVED ___________
DATE FOR RESUBMISSION __________
___________________________
Chairman (Signature)
___________________________________
Printed Name
NIH rDNA category
NOU Number
______________
For EHS use only
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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 vectors)?
Replication
Abortive
If yes, can it cause disease in: - healthy humans?
- Immunocompromised people?
No
No
Yes
Yes
6.
Is immunization required for work at the listed biosafety level?
Is immunization recommended at the listed biosafety level?
Unknown
Unknown
No
No
Yes
Yes
List vaccine:
7.
Is medical surveillance recommended prior to commencement of work?
No
Yes
If yes, please explain (Post exposure management is considered part of occupational health exposure
management)
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8.
Have you educated your staff regarding safe handling and decontamination procedures
for the agents in this NOU?
No
Yes
9.
a.
Risk Assessment:
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
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 the 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:
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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?
BSL1
BSL2
13.
Check the protective clothing or equipment used when handling this agent:
Lab coat/ gloves/ eye protection (BSL2)
PAPR (Racal/Max Air)
N95 respirator
Surgical mask
Face Shield
Safety Centrifuge/blender
Other specify:
Chemical Fume Hood
Biological Safety Cabinet
14.
15.
Room location:
Room location:
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 liquids, then poured down sanitary drain.
Placed in yellow bags for incineration
List disinfectant(s) used for surface decontamination and spills:
Cavicide
Phenolic germicidal
70% alcohol
MicroChem
Other specify:
10% Bleach
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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
If you are planning any animal work, please fill out section III
If you are planning any arthropod vectors work, please fill out section IV
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N/A
N/A
N/A
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:
4.
At what biosafety level will you be conducting the cloning portion (not rescue) of your
research:
BSL1
5.
Room(s):
BSL2
Does this project involve synthetic DNA?
Yes
No
Is this vector/insert commercially purchased?
(No clone manipulation will be done in your lab.)
Please provide the company’s name and product number:
Yes
No
7.
Yes
No
6.
Is this vector/insert provided by a collaborator?
(No clone manipulation will be done in your lab.)
Please provide the collaborator’s institution:
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8.
Please fill out the following table
Source of Insert
DNA/RNA
(Species)
9.
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
Answer the following questions, use additional pages if needed
specify
Additional info needed
a) Does the inserted gene encode a known oncogene?
b) Does the viral DNA integrate into the host genome?
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?
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Yes
No
Yes
No
Yes
No
See section I10:
Evaluation of
Dual use
e) Does the inserted gene have the potential for altering the
cell cycle?
h) Is there a probability of generating replication-competent
viruses?
Yes
No
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
Yes
No
Could this acquisition compromise the use of the drug to control
disease agents in humans, veterinary medicine, or agriculture?
Yes
No
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
A-1a
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?
c) Is the cloning done in Escherichia coli K-12?
Yes
No
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
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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
Yes
No
if no:
work
must be
done at
BSL4
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-3a through III-D-3-d?
Yes
No
D-3-e
Is the DNA from Risk Group 4 agents present in a given recombinant is
totally and irreversibly defective fraction of the agent's genome?
Will this experiment involve DNA from restricted agents to be
transferred into nonpathogenic prokaryotes or lower eukaryotes?
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.
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,
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Yes
No
Yes
No
D-4
D-4-a
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.
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
No
D-4-b
Will this experiment involve the use of whole Plants? Contact EHS
before submitting.
Yes
No
D-5 / E-2
Will this experiment involve more than 10 Liters of culture? Contact
EHS before submitting.
Yes
No
D-6
Experiments with influenza viruses generated by recombinant methods
(e.g., generation by reverse genetics of chimeric viruses with
reassorted segments, introduction of specific mutations) work at the
biosafety level corresponding to the risk group of the virus that was the
source of the majority of segments in the recombinant virus
Yes
No
D-7
Yes
No
D-7
a) Specify biosafety level worked at
Experiments with influenza viruses containing genes or segments from
1918-1919 H1N1 (1918 H1N1), human H2N2 (1957-1968) and highly
pathogenic avian influenza H5N1 strains within the
Goose/Guangdong/96-like H5 lineage (HPAI H5N1 work at BSL3E
Human H2N2 (1957-1968). Experiments with influenza viruses
containing the H2 hemagglutinin (HA) segment - work at BSL3E
Yes
No
D-7a
Human H2N2 (1957-1968). Experiments with the H2 HA gene in
cold-adapted, live attenuated vaccine strains (e.g., A/Ann Arbor/6/60
H2N2) may be conducted at BL2 containment provided segments with
mutations conferring temperature sensitivity and attenuation are not
altered in the recombinant virus. Experiments with Risk Group 2
influenza viruses containing genes from human H2N2 other than the HA
gene - work at BSL2.
Yes
No
D-7a
Highly Pathogenic Avian Influenza H5N1 strains within the
Goose/Guangdong/96-like H5 lineage (HPAI H5N1). Experiments
involving influenza viruses containing a minority/majority of genes
and/or segments from a HPAI H5N1 influenza virus - work at BSL3E
(contact EHS to lower biosafety level)
Yes
No
D-7b
1918 H1N1. Experiments involving influenza viruses containing any
gene or segment from 1918 H1N1- work at BSL3E
Yes
No
D-7c
Yes
No
D-7d (if
yes mark
A1 yes
Antiviral Susceptibility and Containment. – contact EHS before
submission
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An influenza virus containing genes from one of 1918 H1N1, HPAI
H5N1, and human H2N2 (1957-1968) viruses is resistant to both
classes of current antiviral agents, adamantanes and neuraminidase
inhibitors.
also
Experiments with 1918 H1N1, human H2N2 (1957-1968) or HPAI H5N1
that are designed to create resistance to neuraminidase inhibitors or
other effective antiviral agents (including investigational antiviral
agents being developed for influenza)
Will this experiment involve the Formation of Recombinant DNA
Molecules Containing No More than Two-Thirds of the Genome of
any Eukaryotic Virus?
This section covers 1) Recombinant DNA molecules containing no more
than two-thirds of the genome of any eukaryotic virus (all viruses from
a single Family being considered identical) may be propagated and
maintained in cells in tissue culture using BL1 containment. For such
experiments, it must be demonstrated that the cells lack helper virus
for the specific Families of defective viruses being used. 2) The DNA
may contain fragments of the genome of viruses from more than one
Family but each fragment shall be less than two-thirds of a genome.
Yes
No
E-1
Will this experiment involve the use of arthropods?
This section covers experiments with recombinant DNA-modified
arthropods or small animals associated with plants, or with
arthropods or small animals with recombinant DNA-modified
microorganisms associated with them if the recombinant DNAmodified microorganisms have no recognized potential for serious
detrimental impact on managed or natural ecosystems.
Yes
No
E-2-b-5
Will this experiment involve the use of transgenic rodents? This
section covers experiments involving the generation of rodents in which
the animal's genome has been altered by stable introduction of
recombinant DNA, or DNA derived therefrom, into the germ-line
(transgenic rodents).
Yes
No
E-3
Exempt Experiments. The following recombinant DNA molecules are
exempt from the NIH Guidelines and registration with the Institutional
Biosafety Committee is not required.
The following recombinant DNA molecules are exempt: Those that are
not in organisms or viruses.
The following recombinant DNA molecules are exempt: Those that
consist entirely of DNA segments from a single nonchromosomal or
viral DNA source, though one or more of the segments may be a
synthetic equivalent.
The following recombinant DNA molecules are exempt: Those that
consist entirely of DNA from a prokaryotic host including its indigenous
plasmids or viruses when propagated only in that host (or a closely
related strain of the same species), or when transferred to another host
by well established physiological means.
BSL2-Revised 03/12
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F
Yes
No
F-1
Yes
No
F-2
Yes
No
F-3
The following recombinant DNA molecules are exempt: Those that
consist entirely of DNA from an eukaryotic host including its
chloroplasts, mitochondria, or plasmids (but excluding viruses) when
propagated only in that host (or a closely related strain of the same
species).
The following recombinant DNA molecules are exempt: Those that
consist entirely of DNA segments from different species that exchange
DNA by known physiological processes, though one or more of the
segments may be a synthetic equivalent. Contact EHS before
starting.
The following recombinant DNA molecules are exempt: Those that do
not present a significant risk to health or the environment (see Section
IV-C-1-b-(1)-(c), Major Actions), as determined by the NIH Director,
with the advice of the RAC, and following appropriate notice and
opportunity for public comment. See Appendix C, Exemptions under
Section III-F-6 for other classes of experiments which are exempt from
the NIH Guidelines.
Yes
No
F-4
Yes
No
F-5
Yes
No
F-6
If you are planning any animal work, please fill out section III
N/A
If you are planning any arthropod vectors work, please fill out section IV
N/A
BSL2-Revised 03/12
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Section III: Animal Use
A copy of the IACUC flow diagram must be provided.
IACUC Protocol #:
Species:
IACUC Approval date:
1.
Give a project description (specific to animal use, include techniques used per species
and biosampling ½ page):
2.
Can the infected animal present a human health risk after administration?
No
Yes
Route of excretion:
3.
If yes describe provide the following information:
Respiratory
Blood
Milk
Other:
Feces
Saliva
What animal biosafety level is recommended?
ABSL2 for inoculation then housed in ABSL1
4.
Urine
ABSL2
Check the protective clothing level and equipment used when handling the infected
animal. Eye protection to be used at all biosafety levels
ABSL2 PPE
Biological Safety Cabinet
N95 Respirator
Chemical Fume Hood
Other (specify):
5.
Dose per animal (provide volume and concentration):
6.
Route of administration:
Intra-Cranial
Intra-Peritoneal
Intra-Venous
Sub-Cutaneous
Inhalation
Gavage
Other:
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Intra-Dermal
Intra-Cardiac
Topical
13
Intra-Muscular
Intra-Nasal
Oral
7.
8.
Biosampling:
Retro-orbital bleeds
Intra-Venous bleeds
Throat swabs
Other:
Sub-mandibular bleeds
Saphenous bleeds
Urine
Milk
Intra-Cardiac bleeds
Nasal swabs
Feces
Specific animal procedures.
Anesthetic used:
Injectable
Gas- drop method
Anesthetic chamber
Surgeries, specify type of surgery:
Necropsy, specify organs removed:
Perfusions. If checked, please describe the following:
Perfusion system used:
Method of collection of fluids:
Trainer for the laboratory,
including experience and
animal species:
Species:
9.
Will the study use recombinant DNA, viral vectors, gene transfer experiments, or
creation of transgenic animals?
Yes
No
10.
Are the vectors replicative deficient?
11.
Discuss any safety concern associated with the vectors and also are there any toxins or
virulence factors associated with the expression of transgene:
12.
If there is a deviation from standard facility procedures for disposal/treatment of
contaminated materials after usage (animals, bedding, glassware, bench tops, hoods,
etc.), please describe.
Yes
None
BSL2-Revised 03/12
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No
Note: Investigators are reminded that when planning experiments with animals which will
include the use of hazardous agents, you must submit the Hazard Start-Up form located
here: http://research.utmb.edu/arc/forms.shtm
This is required at least 2 weeks prior to startup. All must have approval from the
appropriate committee before you submit the form.
If you are planning any arthropod vector work, please fill out section IV
BSL2-Revised 03/12
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N/A
Section IV: Arthropod Use
1.
Provide a description of the project (Specific to arthropod use):
2.
Arthropod to be used:
Mosquitoes
3.
Indicate life stage used:
Eggs
4.
List animal species to be used:
5.
What Arthropod Containment Level (ACL) is recommended?
ACL2
6.
Check the protective clothing level and equipment used when handling the infected arthropod.
Eye protection to be used at all biosafety level.
Ticks
Larvae
Fleas
Nymphs
Adults
ACL2 PPE
Biological Safety Cabinet
7.
Infection method for arthropod:
Intra-thoracic inoculation
Glove box
Artificial blood meal
Intra-rectal inoculation
BSL2-Revised 03/12
Other specify:
16
Animal feed