INSTITUTIONAL BIOSAFETY COMMITTEE
Tufts University and Tufts Medical Center
Office of the Vice Provost
To submit for review, please e-mail this form to the IBC Office (ibc-office@tufts.edu)
AMENDMENT FORM
Please note that amendments may also affect other aspects of that registration, and those areas should also
be reflected in this document. Some amendments may require submission to the full Institutional Biosafety
Committee. Others may be approved administratively by a Biosafety Officer. The Biosafety Officers reserves the
right to call to full Committee review of the amendment or to require a new registration instead, if necessary.
When submitting an amendment, the Principal Investigator is required to review all of the details of the
original registration to assure the IBC that all unamended details remain identical to the original registration.
Complete this form and send it electronically to ibc-office@tufts.edu to submit for review.
Amendment to Registration #:
Mark the checkboxes to confirm the following:
I am familiar with and agree to abide by the NIH Recombinant DNA Guidelines, CDC/NIH Biosafety
Guidelines, OSHA Standards, institutional policies, and other federal, state and local regulations relating to
this project.
I attest that the information contained in the attached registration is accurate and complete.
I accept responsibility for ensuring that all personnel involved in this project will be trained regarding the
procedures approved, the potential biohazards, relevant biosafety practices, and emergency procedures. I
confirm that the relevant Exposure Response Plan(s) will be followed.
I will submit written reports to the Institutional Biosafety Committee concerning:
1. Any accident that results in a known or potential exposure to recombinant DNA materials, infectious
agents or biological toxins; or any incident resulting in the known or suspected release into the
environment of recombinant DNA materials, infectious agents or biological toxins into the environment.
2. Any problems with physical or biological containment safety procedures or equipment, or facility
failures.
3. Any new information bearing on the safety of this work such as technical data relating to hazard and
safety procedures.
Name of Principal Investigator:
Signature:
Date:
Instructions:
- Please type responses within the space/box provided. You can mark the checkboxes by double-clicking on
the box.
- Submit the completed draft electronically to the IBC Office e-mail (ibc-office@tufts.edu)
- A Biosafety Officer will pre-review the form and determine if the research requires IBC Full Committee
Approval or IBC Administrative Approval. Please be aware that it is in your best interest to submit the draft
to the IBC office well in advance of the submission deadline for the IBC meeting.
I. PROPOSED MODIFICATION(S)
More than one of the Sections below may be affected by the proposed modification. Please complete each
section as necessary.
Addition/change in use of rDNA
Addition/change in use of infectious agents, human material, and biological toxins (exempt amount)
Addition/change in procedure used
Addition or relocation where agent(s) is/are used (any new space must be inspected prior to use)
II. RECOMBINANT DNA
A. Type of Modification
Addition of rDNA (new vectors, etc.)
Change in use of rDNA
B. Justification and Technical Details
1. Purpose of addition/change:
2. Specific DNA/RNA source (or probe):
3. Percent of viral genome in construct:
4. What is being expressed?
5. Do any sequences code for toxins?
6. Identify cloning/expression/transfection vectors used, recipient bacterial strains, and recipient host cell
lines (human, mouse, plant, etc.). Provide a restriction map of vector unless this is a commercially
available vector. If commercially available, please indicate vendor. Describe the location and type of
promoters and other control sequences and percent of any viral genome in construct.
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7. If using viral vectors, indicate packaging cell lines and assay system used to measure helper virus titer or
titer of replication competent virus (background) generated. Include host range of packaged viral vector.
(If using retroviral or lentiviral vectors, additional requirements may apply.)
III. ADDITION OF INFECTIOUS AGENTS, HUMAN MATERIAL, OR BIOLOGICAL TOXINS
A. Type of Modification
Infectious agent(s) to be added:
Human or Non-Human Primate (NHP)
material:
Biological Toxin (exempt amount):
B. Justification and Technical Details
1. Purpose of agent addition:
2. List name(s) of agent(s) and source(s):
3. If the agent is toxic, provide LD50:
4. If the agent is a human pathogen, what is the infectious dose?
5. The risk of exposure will be mitigated by the following: *Check all that apply and indicate with what agent
each is used and the location of use (e.g. lab room #, animal facility, etc.)
Personal Protective Equipment (PPE)
Agent(s) & Location of Use (N/A if not applicable)
Gloves
Lab Coat
Disposable Lab Gown
Disposable Booties
Tyvek Suit
N-95 Respirator
Surgical Mask
PAPR (Powered Air Purifying Respirator)
Safety Glasses
Other:
Engineering Controls (list all that apply)
Biosafety Cabinet
Other:
Date of Certification
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C. Decontamination
If registration contains more than one agent, provide clarification for each of the answers below.
1. Please check
to confirm the information is the same as described in the original registration. If
different answers are applicable for this amendment, please describe below.
2. What chemical and/or thermal process will be used for decontamination?
3. Describe the processes involved in the decontamination and/or disposal of the following:
1. Liquid Waste
2. Contaminated solid waste
3. Cultures, plates, stocks, etc.
4. Animal bedding
(If handled by DLAM/DTRR, please check the box.)
Per standard ABSL-appropriate DLAM/DTRR procedures
5. Animal cages
(If handled by DLAM/DTRR, please check the box.)
Per standard ABSL-appropriate DLAM/DTRR procedures
6. Animal carcasses
(If handled by DLAM/DTRR, please check the box.)
Per standard ABSL-appropriate DLAM/DTRR procedures
IV. ADDITION/CHANGE IN PROCEDURE
A. Procedure to Be Added/Changed
B. Justification and Technical Details
V. PROCEDURES WITH LIVE ANIMALS
Only complete this section if the biohazardous materials will be administered to live animals.
A. Administration of Agent
1. Name of species to be exposed to agent
2. Are the animals immunosuppressed?
No
Yes
3. Agent(s) and doses(s) administered
4. Route(s) of administration
5. Location(s) of administration
Building & Room #
6. Will administration(s) be done inside a
biosafety cabinet?
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7. Will the biological agent or hazardous
metabolite be excreted by urine, feces or
wound drainage? Provide duration, if
applicable.
B. If animals are taken out of the centralized animal facilities, will they be returned to DLAM/DTRR post
administration of the agent(s)?
YES
NO
NOTE: Must follow standard husbandry procedures and cage change frequency requirement.
1. If so, indicate location in centralized animal facility where the animals are returned.
2.
3.
I will use a DTRR/DLAM biohazard cage card to identify the agent used.
I will use an alternative method to identify the biohazardous agent, as described below:
Handling and disposal of bedding, cages and animal carcasses will be done by DTRR/DLAM staff.
Relevant DTRR/DLAM husbandry SOPs will be followed.
Handling and disposal of bedding, cages, and animal carcasses will be done by laboratory staff, as
described below:
VI. LOCATION(S)
List ONLY NEW Laboratories/Facilities where research is to be conducted and the corresponding biosafety
level. Include cold/warm rooms, equipment rooms, and location(s) of biosafety cabinets (BSC). All new spaces
must be inspected by a Biosafety Officer prior to use. This does not apply to locations within the centralized
animal facilities.
Room Number
Room Purpose
(Main Lab, Storage, Tissue Culture, Procedure, etc.)
Biosafety Level
OTHER RELEVANT LINKS
OVP IBC Website:
http://www.tufts.edu/central/research/IBC/index.htm
Tufts University EHS Website:
http://publicsafety.tufts.edu/ehs/
Pathogen Safety Data Sheets:
Biosafety in Microbiological and
Biomedical Laboratories (BMBL):
NIH Recombinant DNA Guidelines:
http://www.phac-aspc.gc.ca/msds-ftss/index.html#menu
http://www.cdc.gov/biosafety/publications/bmbl5/
http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
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APPENDIX A - NIH rDNA Categories
BIOHAZARDOUS MATERIALS REGISTRATION FORM
The NIH rDNA Guidelines identify six categories of experiments involving recombinant DNA:
(i)
those that require Institutional Biosafety Committee (IBC) approval, RAC review, and NIH Director
approval before initiation,
(ii)
those that require NIH/OBA and Institutional Biosafety Committee approval before initiation,
(iii)
those that require Institutional Biosafety Committee and Institutional Review Board approvals and
RAC review before research participant enrollment,
(iv)
those that require Institutional Biosafety Committee approval before initiation,
(v)
those that require Institutional Biosafety Committee notification simultaneous with initiation, and
(vi)
those that are exempt from the NIH Guidelines .
According to the Guidelines, the PI is responsible for identifying which category the proposed rDNA work falls under. The
6 categories are listed and defined below. Mark the box next to the category that you chose.
Section III-F Experiments that are exempt from NIH Guidelines but covered by local regulations. Exempt experiments
must be registered with the Tufts University IBC or the TCSVM IBC as appropriate.
III-F-1
Recombinant DNA molecules that are not in organisms or viruses.
III-F-2
Recombinant DNA molecules 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.
III-F-3
Recombinant DNA molecules 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.
III-F-4
Recombinant DNA molecules that consist entirely of DNA from a eukaryotic host including its
chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or
closely related strain of the same species).
III-F-5
Recombinant DNA molecules 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. See Appendix A-I through A-VI for a list of natural exchangers that are exempt from the “NIH
Guidelines”.
III-F-6
Recombinant DNA experiments that do not present a significant risk to health or the environment as
determined by the NIH Director, RAC and following appropriate notice and opportunity for public
comment. See Appendix C of the NIH Guidelines.
Check the following classes of experiments that are exempt under Section III-F-6 Appendix C
C-1
Recombinant DNA in Tissue Culture?
Recombinant DNA molecules containing less than one-half of any eukaryotic viral genome (all viruses
from a single family being considered identical – see Appendix C-VII-E, Footnotes and References of
Appendix C), that are propagated and maintained in cells in tissue culture are exempt from these NIH
Guidelines with the exceptions listed in Appendix C-1-A.
C-2
Escherichia coli K-12 Host-Vector Systems?
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Experiments which use Escherichia coli K-12 host-vector system, with the exception of those experiments
listed in Appendix C-II-A are exempt from the NIH Guidelines provided that: (i) the Escherichia coli host
does not contain conjugation proficient plasmids or generalized transducing phages; or (ii) lambda or
lambdoid of Ff bacteriophages or non-conjugative plasmids (see Appendix c-VI-B, Footnotes and
References of Appendix C) shall be used as vectors. However, experiments involving the insertion into
Escherichia coli K-12 of DNA from prokaryotes that exchange genetic information (see Appendix C-VI-C,
Footnotes and References of Appendix C) with Escherichia coli may be performed with any Escherichia coli
K-12 vector (e.g., conjugative plasmid). When a non-conjugative-proficient vector is used, the Escherichia
coli K-12 host may contain conjugation-proficient plasmids either autonomous or integrated, or
generalized transducing phages. For these exempt laboratory experiments, Biosafety Level 1 (BSL1)
physical containment conditions are recommended. For large scale fermentation experiments, the
appropriate physical containment conditions need not be greater than those for the host organism
unmodified by recombinant DNA techniques; the Institutional Biosafety Committee can specify higher
containment if deemed necessary.
C-III
Saccharomyces Host-Vector Systems?
Experiments involving Saccharomyces cerevisiae and Saccharomyces uvarum host-vector systems, with
the exception listed in Appendix C-III-A, are Exempt from the NIH Guidelines. For these exempt
experiments, BSL1 physical containment is recommended. For large scale fermentation experiments, the
appropriate physical containment conditions need be no greater than those for the host organism
unmodified by recombinant DNA techniques; the Institutional Biosafety Committee can specify higher
containment if deemed necessary.
C-IV
Kluyveromyces Host-Vector Systems
Experiments involving Kluyveromyces lactis host-vector systems, with the exception of experiments listed
in Appendix C-IV-A, are exempt from the NIH Guidelines provided laboratory-adapted strains are used (i.e.
strains that have been adapted to growth under optimal or defined laboratory conditions). For these
exempt experiments, BL1 physical containment is recommended. For large-scale fermentation
experiments, the appropriate physical containment conditions need be no greater than those for the host
organism unmodified by recombinant DNA techniques; the Institutional Biosafety Committee may specify
higher containment if deemed necessary.
C-V
Bacillus subtilis or Bacillus licheniformis Host-Vector Systems?
Any asporogenic Bacillus subtilis or asporogenic Bacillus licheniformis strain which does not revert to a
spore-former with a frequency greater than 107 may be used for cloning DNA with the exception of those
experiments listed in Appendix C-IV-A, Exceptions. For these exempt laboratory experiments, BSL1
physical containment conditions are recommended. For large scale fermentation experiments, the
appropriate physical containment conditions need be no greater than those for the host organism
unmodified by recombinant DNA techniques; the Institutional Biosafety Committee can specify higher
containment if deemed necessary.
Recombinant DNA molecules derived entirely from extrachromosomal elements of the organism listed
below (including shuttle vectors constructed from vectors described in Appendix C), propagated and
maintained in organisms listed below are exempt from these NIH Guidelines.
Appendix C-VI. Extrachromosomal Elements of Gram Positive Organisms
Recombinant DNA molecules derived entirely from extrachromosomal elements of the organisms listed
below (including shuttle vectors constructed from vectors described in Appendix C), propagated and
maintained in organisms listed below are exempt from these NIH Guidelines.
C-VII
The Purchase or Transfer of Transgenic Rodents?
The purchase or transfer of transgenic rodents for experiments that require ABSL1 containment (see
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Appendix G-III-M, Footnotes and References of Appendix G) are exempt from the NIH Guidelines.
Appendix C-VIII. Generation of BL1 Transgenic Rodents via Breeding
The breeding of two different transgenic rodents or the breeding of a transgenic rodent and a nontransgenic rodent with the intent of creating a new strain of transgenic rodent that can be housed at BL1
containment will be exempt from the NIH Guidelines if: (1) Both parental rodents can be housed under
BL1 containment; and
(2) neither parental transgenic rodent contains the following genetic modifications: (i) incorporation of
more than one-half of the genome of an exogenous eukaryotic virus from a single family of viruses; or (ii)
incorporation of a transgene that is under the control of a gammaretroviral long terminal repeat (LTR);
and
(3) the transgenic rodent that results from this breeding is not expected to contain more than one-half of
an exogenous viral genome from a single family of viruses.
Section III-E Experiments that require IBC notice simultaneous with initiation.
III-E
Experiments not included in Sections III-A, III-B, III-C, III-D, III-F and their subsections are considered in
this section. All such experiments may be conducted at BSL 1.
III-E-1
Experiments involving the formation of r-DNA molecules containing no more than 2/3 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 BSL 1 containment. It must be shown that the cells lack helper
virus for the specific Families of defective viruses used. The DNA may contain fragments of the genome
of viruses from more than one Family, but each fragment shall be less than 2/3 of a genome.
III-E-2
Experiments involving r-DNA-modified whole plants, and/or experiments involving r-DNA-modified
organisms associated with plants, except those that fall under Section III-A, III-B, III-C, III-D, or III-F. See
Section III-E-2 for recommendation of containment levels.
III-E-3
Experiments involving the generation of rodents in which the animal’s genome has been altered by
stable introduction of recombinant DNA, or DNA derived there from, into the germ-line (transgenic
rodents). Only experiments that require BSL1 containment are covered under this section; experiments
that require BSL2, BSL3, or BSL4 containment are covered under Section III-D-4.
Section III-D Experiments that require IBC approval before initiation of experiments.
III-D-1
Experiments using Risk Group 2, Risk Group 3, Risk Group 4, or restricted agents as host-vector systems.
(See Appendix B for information on Risk Groups.)
III-D-1-a
Introduction of recombinant DNA into Risk Group 2 (RG-2) agents is usually conducted at BSL2
containment. Experiments with such agents will usually be conducted with whole animals at BSL2 or
ABSL-2 containment. (See Appendix B for information on Risk Groups.)
III-D-1-b
Introduction of recombinant DNA into Risk Group 3 (RG-3) agents is usually conducted at BSL3
containment. Experiments with such agents will usually be conducted with whole animals at BSL3 or
ABSL-3 containment. (See Appendix B for information on Risk Groups.)
III-D-1-c
Introduction of recombinant DNA into Risk Group 4 (RG-4) agents is usually conducted at BSL4
containment. Experiments with such agents will usually be conducted with whole animals at BSL4 or
ABSL-4 containment. (See Appendix B for information on Risk Groups.)
III-D-2
Experiments in which DNA from Risk Group 2, Risk Group 3, Risk Group 4, or restricted agents is cloned
into nonpathogenic prokaryotic or lower eukaryotic host-vector systems.
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III-D-2-a
Experiments in which DNA from RG- 2 or RG- 3 agents is transferred into nonpathogenic prokaryotes or
lower eukaryotes may be conducted at BSL2 containment. Experiments in which DNA from RG-4 agents
is transferred into nonpathogenic prokaryotes or lower eukaryotes may be performed under BSL2
containment after demonstration that only a totally and irreversibly defective fraction of the agent’s
genome is present in a given recombinant. The IBC may approve the specific lowering of containment
for particular experiments to BSL1. Many experiments in this category are exempt from the “NIH
Guidelines”. (See Appendix B for information on Risk Groups.)
III-D-2-b
Experiments in which DNA from restricted agents is transferred into nonpathogenic prokaryotes or
lower eukaryotes shall be performed under containment conditions determined by NIH/OBA following a
case-by-case review.
III-D-3
Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses in the
presence of helper virus in tissue culture systems.
III-D-3-a
Experiments involving the use of infectious or defective RG-2 viruses in the presence of helper virus may
be conducted at BSL2 containment. (See Appendix B for information on Risk Groups.)
III-D-3-b
Experiments involving the use of infectious or defective RG-3 viruses in the presence of helper virus may
be conducted at BSL3 containment. (See Appendix B for information on Risk Groups.)
III-D-3-c
Experiments involving the use of infectious or defective RG-4 viruses in the presence of helper virus may
be conducted at BSL4 containment. (See Appendix B for information on Risk Groups.)
III-D-3-d
Experiments involving the use of infectious or defective restricted poxviruses in the presence of helper
virus shall be determined on a case-by-case basis following NIH/OBA review. A USDA permit is required
for work with plant or animal pathogens.
III-D-3-e
Experiments involving 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 may be conducted at BSL1.
III-D-4
Experiments involving whole animals.
III-D-4-a
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 and propagated under conditions of physical containment comparable to BSL1 or
ABSL1 and appropriate to the organism under study. 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, may be propagated under conditions of physical containment comparable
to BSL1 or ABSL1 and appropriate to the organism under study. Experiments involving the introduction
of other sequences from eukaryotic viral genomes into animals are covered under Section III-4-b. The
investigator must demonstrate that the fraction of the viral genome being utilized does not lead to
productive infection.
III-D-4-b
Experiments involving recombinant DNA, or DNA or RNA derived therefrom, involving whole animals,
including transgenic animals, and not covered by Sections III-D-1 or III-D-4-a, may be conducted at the
appropriate containment determined by the IBC.
III-D-4-c-1
Experiments involving the generation of transgenic rodents that require BSL1 containment are described
under Section III-E-3.
III-D-4-c-2
Purchase or transfer of transgenic rodents is exempt from the “NIH Guidelines” under Section III-F.
III-D-5
Experiments involving whole plants. Experiments to genetically engineer plants by R-DNA methods, to
use plants for other experimental purposes, to propagate such plants, or to use plants together with
microorganisms or insects containing R-DNA, may be conducted under the containment conditions
described in the “NIH Guidelines”, Section III-D-5a through Section III-D-5e.
III-D-6
Experiments involving more than 10 liters of culture. The appropriate containment will be decided by
the IBC. Where appropriate, Appendix K of the “NIH Guidelines” will be used to determine containment
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conditions.
III-D-7
Experiments Involving Influenza Viruses. Experiments with influenza viruses generated by recombinant
methods (e.g., generation by reverse genetics of chimeric viruses with reassorted segments,
introduction of specific mutations) shall be conducted at the biosafety level containment corresponding
to the risk group of the virus that was the source of the majority of segments in the recombinant virus
(e.g., experiments with viruses containing a majority of segments from a RG3 virus shall be conducted at
BL3). 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) shall be conducted at BL3 enhanced containment
(see Appendix G-II-C-5, Biosafety Level 3 Enhanced for Research Involving Risk Group 3 Influenza
Viruses) unless further noted in the Guidelines.
Section III-C Experiments that require IBC and Institutional Review Board (IRB) approvals, and RAC review before
research participant enrollment.
III-C-1
Experiments involving the deliberate transfer of (1) recombinant DNA, or (2) DNA or RNA derived from
recombinant DNA, into one or more human subjects.
Section III-B Experiments that require NIH/OBA and IBC approval before initiation.
III-B-1
Experiments involving the cloning of toxin molecules with LD50 of less than 100 nanograms per kilogram
body weight.
Section III-A Experiments that require Institutional Biosafety Committee (IBC) approval, Recombinant DNA Advisory
Committee (RAC) review, and NIH Director approval before initiation of experiments.
III-A-1-a.
Deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait
naturally, if such acquisition could compromise the use of the drug to control disease agents in humans,
veterinary medicine or agriculture.
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