Frequently Asked Questions:
Biological Safety Practices
for Research with Potentially
Highly Pathogenic Influenza Virus Strains
NIH Office of Biotechnology Activities - Biological Safety Practices for Research with Potentially Highly Pathogenic Influenza Virus Strains
Frequently Asked Questions:
Biological Safety Practices for Research with
Potentially Highly Pathogenic Influenza Virus Strains
The National Institutes of Health (NIH) Office of Biotechnology Activities (OBA) periodically
receives questions about the appropriate containment for various types of influenza strains used in
recombinant DNA research. This is especially true for non-contemporary human influenza strains
(including but not limited to H2N2 and 1918 influenza virus) and circulating highly pathogenic
avian influenza (HPAI) viruses. The following frequently asked questions are intended to assist
investigators and Institutional Biosafety Committees (IBCs) in making decisions about
containment and biosafety practices for research with these pathogenic strains or their genetic
elements.
1. What regulations and guidelines pertain to research with non-contemporary strains of
influenza virus?
The principal policies addressing the biosafety and biosecurity considerations relating to
research with non-contemporary strains of influenza are the:
 NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH
Guidelines);
 Department of Health and Human Services (HHS) Centers for Disease Control and
Prevention (CDC) and US Department of Agriculture Animal (USDA) Animal and
Plant Health Inspection Service (APHIS) Select Agent Rules; and
 CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories
(BMBL) 5th Edition.
Certain NIH Institutes and Centers may have their own terms and conditions of grant award in
addition to the standards articulated by the policies above (see question 5 below).
2. What should be considered when conducting a risk assessment for research with noncontemporary strains of influenza?
The NIH Guidelines address research with influenza viruses to the extent they have been
genetically modified with recombinant DNA or are being used with transgenic animals. The
NIH Guidelines describe four levels of Risk Groups into which infectious agents are placed in
accordance with their potential impact on human health, as well as four biological safety
levels, in keeping with the risk of the research. Appendix B of the NIH Guidelines lists
Orthomyxoviruses (influenza) types A, B, and C as Risk Group 2 agents. Users of the NIH
Guidelines should understand, however, that Risk Group assignments are just the starting point
for the risk assessment process, and for the determination of the appropriate biosafety level at
which to conduct research with pathogenic agents.
A comprehensive risk assessment of research involving a potentially highly
pathogenic influenza strain would generally lead to the conclusion that the work
should be conducted at Biosafety Level 3 or higher.
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NIH Office of Biotechnology Activities - Biological Safety Practices for Research with Potentially Highly Pathogenic Influenza Virus Strains
As noted in Section II-A-3 of the NIH Guidelines, risk assessments should include a thorough
consideration of such factors as virulence, pathogenicity, infectious dose, environmental
stability, route of spread, communicability, quantity, availability of a vaccine or treatment,
toxicity, physiological activity, and allergenicity. Any strain that is known to be more
hazardous than the parent (wild-type) strain should be considered for handling at a higher
containment level than typically assigned to its Risk Group.
In deciding on the appropriate containment for an experiment involving non-contemporary
strains of influenza virus, an initial risk assessment must be conducted based on the known and
potential properties of the agent. This should be followed by a thorough consideration of how
the agent is to be manipulated.
Important considerations in working with these strains include:
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The number of years since an antigenically related virus last circulated and the potential
for presence of a susceptible population;
Whether an efficacious vaccine is available and whether the agent is likely to be
susceptible to current antivirals;
The gene constellation used;
Whether there is clear evidence of reduced virus replication in the respiratory tract of
appropriate animal models, compared with the level of replication of the wild-type
parent virus from which it was derived; and
The degree of clonal purity and phenotypic stability.
In general, non-contemporary human influenza virus strains, HPAI viruses,
and chimeras containing genetic elements from highly pathogenic influenza
virus strains present more significant biosafety and public health
considerations than seasonal influenza strains and should be contained under
Biosafety Level 3 conditions. More stringent biosafety controls may be
appropriate, depending on the outcome of the risk assessment. If adequate
risk assessment data are not available, a more cautious approach utilizing
higher biosafety levels and additional practices is warranted.
3. What are the current CDC/NIH recommendations for working with non-contemporary
strains of influenza viruses?
The BMBL recommends Biosafety Level 3 and Animal Biosafety Level 3 practices,
procedures and facilities with rigorous adherence to clothing change protocols and additional
respiratory protection such as that afforded by negative pressure, HEPA-filtered respirators or
positive air-purifying respirators (PAPRs).
Any research involving reverse genetics of the 1918 influenza strain in particular should
proceed with extreme caution. The risk to laboratory workers is unknown at the present time,
but the pandemic potential is thought to be significant. Until further risk assessment data are
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NIH Office of Biotechnology Activities - Biological Safety Practices for Research with Potentially Highly Pathogenic Influenza Virus Strains
available, the following practices and conditions at a minimum are recommended for
manipulation of these viruses and for the use of laboratory animals infected with these viruses:
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Biosafety Level 3 and Animal Biosafety Level 3 practices, procedures and facilities;
Large laboratory animals such as nonhuman primates should be housed in primary
barrier systems in Animal Biosafety Level 3;
Rigorous adherence to additional respiratory protection and clothing change protocols;
Use of negative pressure, high efficiency particulate air (HEPA)-filtered respirators or
positive air-purifying respirators (PAPRs);
Use of HEPA filtration for treatment of exhaust air; and
Amendment of personnel practices to include personal showers prior to exiting the
laboratory.
4. What are the specific biosafety recommendations for handling Highly Pathogenic Avian
Influenza (HPAI) viruses?
Manipulating HPAI viruses in biomedical research laboratories requires extreme caution
because some strains may pose increased risk to laboratory workers and have significant
agricultural and economic consequences. The BMBL recommends a minimum of Biosafety
Level 3 and Animal Biosafety Level 3 practices, procedures and facilities along with clothing
change and personal showering protocols. Negative pressure, HEPA-filtered respirators or
positive air-purifying respirators are recommended for HPAI viruses with potential to infect
humans.
Detailed biosafety recommendations for work with various human and animal influenza
viruses, including HPAI viruses, can be found in the agent summary statement for influenza
virus in the 5th edition of the BMBL. Readers should also be aware that HPAI viruses are
agricultural Select Agents requiring registration under the USDA/APHIS Select Agent rule.
All work with HPAI strains of influenza (including non-HPAI) requires an APHIS-issued
permit. In addition, other strains of avian influenza are covered by USDA regulations
pertaining to the interstate transport of avian influenza viruses generally.
5. Are there are particular terms and conditions of award that NIH places on work with
non-contemporary strains of influenza viruses?
As stated in Section I-D of the NIH Guidelines and reiterated in the NIH Grants Policy Manual,
as a condition for NIH funding of recombinant DNA research, institutions shall ensure that
such research conducted at or sponsored by the institution, irrespective of the source of
funding, shall comply with the NIH Guidelines.
NIH Institutes and Centers may have their own terms and conditions of grant award in addition
to the standards articulated by the BMBL and the NIH Guidelines. You should consult with
your NIH program officer for information on any such terms and conditions.
6. Are there are special occupational health considerations that should be considered when
working with non contemporary human strains of influenza viruses, including
recombinants and reassortants; with HPAI viruses; or with viruses created by reverse
genetics of the 1918 strain?
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NIH Office of Biotechnology Activities - Biological Safety Practices for Research with Potentially Highly Pathogenic Influenza Virus Strains
According to the BMBL, institutions performing work with HPAI viruses; non-contemporary
human influenza strains, including recombinants and reassortants; and viruses created by
reverse genetics of the 1918 pandemic strain should consider developing and implementing a
specific medical surveillance and response plan. These plans may include:
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Storage of baseline serum samples from individuals working with these influenza strains;
Annual vaccination with the currently licensed influenza vaccine for such individuals;
Providing employee counseling regarding disease symptoms including fever, conjunctivitis
and respiratory symptoms;
Establishing a protocol for monitoring personnel for these symptoms; and
Establishing a clear medical protocol for responding to suspected laboratory-acquired
infections. Antiviral drugs (e.g., oseltamivir, amantadine, rimantadine, zanamivir) should
be available for treatment and prophylaxis, as necessary. It is recommended that the
sensitivities of the virus being studied to the antivirals be ascertained.
In addition, all personnel should be enrolled in an appropriately constituted respiratory
protection program.
7. Where can I find out more information on conducting a risk assessment with noncontemporary influenza strains?
Information on risk assessments may be found in the NIH Guidelines for Research Involving
Recombinant DNA Molecules Section II: Safety Considerations.
Information on agent risk assessments may be found in the Agent Summary Statements of the
CDC/NIH publication Biosafety in Microbiological and Biomedical Laboratories.
Further guidance on agents may be obtained through:
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National Institutes of Health Office of Biotechnology Activities
Phone: 301-496-9838
Email: oba@od.nih.gov
Web Site: http://www4.od.nih.gov/oba
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Centers for Disease Control and Prevention Select Agent Program
Phone: 404-718-2000
Email: lrsat@cdc.gov
Web Site: http://www.cdc.gov/od/sap/
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United States Department of Agriculture Animal and Plant Health Inspection Service,
Select Agent Program
Phone: 301-734-5960
Email: Agricultural.Select.Agent.Program@aphis.usda.gov
Web Site: http://www.aphis.usda.gov/programs/ag_selectagent/
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