81 FR 22286 - National Institutes of Health (NIH) Office of Science Policy (OSP) Recombinant or Synthetic Nucleic Acid Research: Action Under the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines)

DEPARTMENT OF HEALTH AND HUMAN SERVICES
National Institutes of Health

Federal Register Volume 81, Issue 73 (April 15, 2016)

The NIH OSP is amending portions of the NIH Guidelines in order to provide investigators with biosafety guidance regarding the standards for containment of non-human primates (NHPs) in biosafety level (BL) 4 laboratories and to make such guidance consistent with the expectations articulated in the Centers for Disease Control and Prevention (CDC)/NIH Biosafety in Microbiological and Biomedical Laboratories 5th edition (BMBL). Specifically, the NIH Guidelines will allow for housing of NHPs in open caging in a dedicated animal holding room provided there are two physical barriers between that animal holding room and non-containment space within the laboratory, the animal holding room has negative air pressure with respect to any adjacent non-containment corridors, and there are specific decontamination protocols in place before the door to the animal holding room is opened to allow for the periodic transfer of new animals into the room. These amendments do not change the current containment requirements in the NIH Guidelines but rather offer an alternative for achieving primary containment without compromising safety. In addition, the recertification requirement for biosafety cabinets in BL4 laboratories is updated in recognition of the technological standards for modern biosafety cabinets. The NIH OSP also is updating the validation requirements for equipment responsible for centralized heat decontamination of liquid effluents in laboratories working with large animals. These amendments to the NIH Guidelines will be implemented immediately upon publication in the Federal Register. These changes were developed after extensive consultation with biosafety experts, directors of and principal investigators in BL4 facilities working with NHPs, and CDC's Division of Select Agent and Toxins (DSAT) leadership at a public workshop and discussion at a public Recombinant DNA Advisory Committee (RAC) meeting. Publication in the Federal Register will inform the scientific and biosafety communities.

[Federal Register Volume 81, Number 73 (Friday, April 15, 2016)]
[Notices]
[Pages 22286-22289]
From the Federal Register Online  [www.thefederalregister.org]
[FR Doc No: 2016-08810]


-----------------------------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES

National Institutes of Health


National Institutes of Health (NIH) Office of Science Policy 
(OSP) Recombinant or Synthetic Nucleic Acid Research: Action Under the 
NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic 
Acid Molecules (NIH Guidelines)

SUMMARY: The NIH OSP is amending portions of the NIH Guidelines in 
order to provide investigators with biosafety guidance regarding the 
standards for containment of non-human primates (NHPs) in biosafety 
level (BL) 4 laboratories and to make such guidance consistent with the 
expectations articulated in the Centers for Disease Control and 
Prevention (CDC)/NIH Biosafety in Microbiological and Biomedical 
Laboratories 5th edition (BMBL). Specifically, the NIH Guidelines will 
allow for housing of NHPs in open caging in a dedicated animal holding 
room provided there are two physical barriers between that animal 
holding room and non-containment space within the laboratory, the 
animal holding room has negative air pressure with respect to any 
adjacent non-containment corridors, and there are specific 
decontamination protocols in place before the door to the animal 
holding room is opened to allow for the periodic transfer of new 
animals into the room. These amendments do not change the current 
containment requirements in the NIH Guidelines but rather offer an 
alternative for achieving primary containment without compromising 
safety.
    In addition, the recertification requirement for biosafety cabinets 
in BL4 laboratories is updated in recognition of the technological 
standards for modern biosafety cabinets. The NIH OSP also is updating 
the validation requirements for equipment responsible for centralized 
heat decontamination of liquid effluents in laboratories working with 
large animals.
    These amendments to the NIH Guidelines will be implemented 
immediately upon publication in the Federal Register. These changes 
were developed after extensive consultation with biosafety experts, 
directors of and principal investigators in BL4 facilities working with 
NHPs, and CDC's Division of Select Agent and Toxins (DSAT) leadership 
at a public workshop and discussion at a public Recombinant DNA 
Advisory Committee (RAC) meeting. Publication in the Federal Register 
will inform the scientific and biosafety communities.

FOR FURTHER INFORMATION CONTACT: If you have questions, or require 
additional information about these changes, please contact the NIH OSP 
by email at [email protected], or telephone at 301-496-9838.

SUPPLEMENTARY INFORMATION: The first three editions of the BMBL and the 
NIH Guidelines were consistent in their approach to requiring primary 
containment for animal work in BL4 containment laboratories. However, 
in the early 1990s, the BMBL was amended and the fourth edition stated 
that animals housed in BL4 suit laboratories (i.e., laboratories in 
which Class III cabinets are not used but instead personnel wear 
positive pressure protective suits) should be housed in a primary 
containment system (such as open cages covered with filtered bonnets 
and opened in laminar flow hoods or other equivalent containment 
systems). This language remains in the current BMBL (5th edition). With 
the change in the BMBL, primary containment caging was arguably 
preferred but no longer required under BL4 containment. In contrast, 
the NIH Guidelines have always required primary containment caging for 
all animals in BL4 laboratories.
    Non-human primates are social animals and require environmental 
enrichment. Researchers in several U.S. BL4 laboratories engaged in NHP 
research approached the NIH OSP with concerns that primary containment 
caging in BL4 laboratories hindered the creation of an environment that 
allowed animals to benefit from adequate social interaction. Also based 
on risk assessments and experiences comparing several primary 
containment caging systems, the researchers concluded that primary 
containment caging may actually create new hazards for laboratory 
workers. These findings included interference with observation of the 
animals from outside the room leading to more frequent entries into the 
BL4 animal room to monitor the animals, and exacerbation of cramped 
working conditions created by the additional barriers required by some 
containment systems, which increases the difficulty of working in 
inflated pressure suits as well as the potential for damage to the 
pressure suit. In addition, investigators stated that current BL4 
laboratory designs incorporate sophisticated engineering systems, which 
provide biosafety protection in a dedicated animal room equivalent to 
the primary containment caging required under the NIH

[[Page 22287]]

Guidelines. On March 28, 2014, the NIH OSP, together with the CDC 
Division of Select Agents (DSAT), held a meeting with investigators and 
biosafety personnel from nine BL4 laboratories in the U.S. and five 
international laboratories (agenda and roster available at http://osp.od.nih.gov/office-biotechnology-activities/event/2014-03-28-120000-2014-03-28-211500/primary-containment-non-human-primates-biosafety-level-4-laboratories-challenges-and-best-practices). That meeting was 
followed by a discussion at the June 11, 2014, meeting of the NIH RAC 
regarding housing of NHPs in BL4 laboratories (a webcast of that 
discussion is available at http://videocast.nih.gov/summary.asp?Live=14300&bhcp=1.)
    As a result of these discussions and after consultation with CDC 
DSAT, the NIH OSP is amending the NIH Guidelines to allow the housing 
of NHP in open cages in a dedicated animal room provided certain 
conditions as articulated below are met.
    In addition, OSP is updating the requirement for testing and 
certification of Class I and II biosafety cabinets at BL4 from every 
six months to annually, recognizing the technological advances in 
biosafety cabinet design and engineering that have occurred over the 
period since this performance measure was originally implemented.
    Finally, Appendix Q of the NIH Guidelines currently requires that 
the centralized heat treatment catch tank system in BL4 large animal 
laboratories be validated every 30 days. To carry out this testing 
effectively, the system should be near capacity (i.e. ``under load''). 
Some laboratories do not use their catch tank systems on a regular 
basis; therefore, a mandatory 30 day validation would require them to 
demonstrate that the equipment is functioning when it is not in use or 
is not at capacity. This will not serve the intended purpose of 
demonstrating that the equipment is functioning as intended. Therefore, 
validation intervals will need to be set based on the utilization of 
the system provided it is done at least once a year.
    To implement these changes, the following sections of the NIH 
Guidelines are to be amended:
    Appendix G-II-D-2-l. currently states:

    Appendix G-II-D-2-l. Laboratory animals involved in experiments 
requiring BL4 level physical containment shall be housed either in 
cages contained in Class III cabinets or in partial containment 
caging systems, such as Horsfall units (see Appendix G-III-K, 
Footnotes and References of Appendix G), open cages placed in 
ventilated enclosures, or solid-wall and -bottom cages placed on 
holding racks equipped with ultraviolet irradiation lamps and 
reflectors that are located in a specially designed area in which 
all personnel are required to wear one-piece positive pressure 
suits.

    Appendix G-II-D-2-1 is amended as follows with the addition of a 
new Appendix G-II-D-2-1-(2) that will address housing of NHPs:

Appendix G-II-D-2-l. Containment for Animal Research

    Appendix G-II-D-2-l-(1). Laboratory animals involved in experiments 
requiring BL4 level physical containment shall be housed either in 
cages contained in Class III cabinets or in partial containment caging 
systems, such as Horsfall units (see Appendix G-III-K, Footnotes and 
References of Appendix G), open cages placed in ventilated enclosures, 
or solid-wall and -bottom cages placed on holding racks equipped with 
ultraviolet irradiation lamps and reflectors that are located in a 
specially designed area in which all personnel are required to wear 
one-piece positive pressure suits.
    Appendix G-II-D-2-l-(2). Non-human primates (NHP) may be housed (1) 
under the containment conditions described in Appendix G-II-D-2-l-(1) 
above, or (2) in open cages within a dedicated animal holding room that 
serves as the primary barrier and in which all personnel are required 
to wear one-piece positive pressure suits. A room serving as a primary 
barrier must be air-tight and capable of being decontaminated using 
fumigation. If NHPs are to be contained in a dedicated animal holding 
room serving as the primary barrier, the following conditions shall be 
met:
    (i) Access to the animal holding room from service corridors 
outside of the BL4 containment space shall require passage through two 
sets of doors, and the inner most door must be an air pressure 
resistant (APR) door;
    (ii) For any animal holding room considered to be a primary 
barrier, APR door(s) providing direct ingress from the exterior service 
corridor shall be fitted with appropriate and redundant lock-out 
mechanisms to prevent access when the animal holding room is 
contaminated and in use. There should be more than one mechanism to 
ensure that this primary barrier door cannot be opened when the animal 
room is contaminated and the APR door shall not serve as an emergency 
exit from the BL4 laboratory. The APR door shall be appropriately 
tested to demonstrate that in the closed, locked-out mode, the door 
provides an air-tight barrier proven by pressure decay testing or other 
equivalent method;
    (iii) Any door(s) allowing access into a corridor from which there 
is direct ingress to an animal holding room must be fitted with either 
(1) an APR door or (2) a non-APR door, providing directional airflow is 
maintained from the laboratory corridor space into the animal room. For 
the purpose of fumigation, animal rooms equipped with non-APR doors 
opening into the adjacent interior corridors shall be considered one 
space (i.e., areas between air-tight doors shall be fumigated 
together).
    (iv) Any door(s) used for access to the service corridor (the 
secondary barrier) shall be self-closing and of solid construction, 
designed not to corrode, split or warp;
    (v) Access to the service corridor inside the secondary barrier 
shall be restricted and strictly controlled when animal rooms are in 
use. Whenever possible, the secondary barrier door(s) should be fitted 
with safety interlock switches designed to prevent it from opening when 
an animal holding room door (the primary barrier) is opened following 
room decontamination; if interlock devices cannot be used, specific 
administrative procedures shall be implemented to control access to the 
service corridor;
    (vi) The service corridor shall maintain a negative pressure 
(inward directional airflow) relative to adjoining traffic corridors;
    (vii) Prior to fumigation of the animal holding room, cages should 
be removed for autoclaving or chemical decontamination.
    (viii) Caging should be chosen to reduce the amount of animal 
detritus that can be thrown out of the cage and onto the floor of the 
animal holding room;
    (ix) The flow of personnel, material and equipment should be 
directed in order to minimize the spread of contamination from the 
animal holding room into adjacent areas of the laboratory.
    (x) Following animal room decontamination, safeguards involving the 
use of personal protective equipment and appropriate administrative 
controls shall be implemented for the safe retrieval of biological 
indicators in order to prevent the spread of infectious agents in the 
event of a decontamination failure.
    With regard to the frequency of class II biosafety cabinet 
recertification and testing, the NIH Guidelines require 
recertification/testing of biosafety cabinets at six-month intervals. 
However, modern biosafety cabinet

[[Page 22288]]

design specifications incorporate continuous electronic and physical 
monitoring systems that track multiple operational parameters such as 
pressure differential, air flow velocity and plenum pressure with added 
capabilities for remote control and monitoring. Continuous performance 
monitoring and redundant safety features obviate the need for frequent 
testing of modern biosafety cabinets under normal conditions of use. 
Also the testing of biosafety cabinets in biomedical high containment 
(BL4) laboratories entails a complete shutdown of the laboratory for 
decontamination with appropriate sterilants to allow technicians to 
access the equipment safely. This procedure increases risks to 
laboratory staff working with the sterilants (often in the form of 
toxic gasses) and requires a halt to all research activities for an 
extended period of time depending on the number of cabinets to be 
tested or recertified. The NIH is in agreement with the recommendation 
of the BMBL that class II biosafety cabinets be tested and certified at 
least annually, with the understanding that retesting may have to be 
performed as needed at the discretion of the Institutional Biosafety 
Committee (IBC), if for example, equipment is moved or subject to 
unusual conditions of use.
    Appendix G-II-D-4-p currently states:

    Appendix G-II-D-4-p. The treated exhaust air from Class I and II 
biological safety cabinets may be discharged into the laboratory 
room environment or the outside through the facility air exhaust 
system. If exhaust air from Class I or II biological safety cabinets 
is discharged into the laboratory the cabinets are tested and 
certified at six-month intervals. The exhaust air from Class III 
biological safety cabinets is discharged, without recirculation 
through two sets of high efficiency particulate air/HEPA filters in 
series, via the facility exhaust air system. If the treated exhaust 
air from any of these cabinets is discharged to the outside through 
the facility exhaust air system, it is connected to this system in a 
manner (e.g., thimble unit connection (see Appendix G-III-L, 
Footnotes and References of Appendix G)) that avoids any 
interference with the air balance of the cabinets or the facility 
exhaust air system.

    Appendix G-II-D-4-p is amended as follows:
    Appendix G-II-D-4-p. The treated exhaust air from Class I and II 
biological safety cabinets may be discharged into the laboratory room 
environment or the outside through the facility air exhaust system. If 
exhaust air from Class I or II biological safety cabinets is discharged 
into the laboratory the cabinets are tested and certified at minimum on 
a yearly basis. More frequent testing and certification, based on the 
amount of use or other safety factors, shall be left to the discretion 
of the IBC. The exhaust air from Class III biological safety cabinets 
is discharged, without recirculation through two sets of high 
efficiency particulate air/HEPA filters in series, via the facility 
exhaust air system. If the treated exhaust air from any of these 
cabinets is discharged to the outside through the facility exhaust air 
system, it is connected to this system in a manner (e.g., thimble unit 
connection (see Appendix G-III-L, Footnotes and References of Appendix 
G)) that avoids any interference with the air balance of the cabinets 
or the facility exhaust air system.
    With regard to the periodic biologic validation of the centralized 
heat treatment process for all BL3 and BL4 facilities, the NIH 
Guidelines requires that this process be performed every 30 days while 
the BMBL recommends at least an annual biological validation. Similar 
to biosafety cabinets, modern catch tank systems for heat treatment of 
all laboratory effluents have also become more sophisticated and now 
incorporate redundant monitoring systems to track temperature and 
pressure parameters during each heat treatment cycle. In addition it 
should be noted that proper validation of the heat treatment catch tank 
system requires that testing be performed when the system is at or near 
capacity (i.e. ``under load'')--more frequent validation of a heat 
treatment system that is below capacity may not serve the intended 
purpose of demonstrating that the equipment is functioning as intended. 
An additional margin of safety is achieved by monitoring sterilization 
cycle parameters on a routine basis. We are, therefore, in agreement 
with the recommendation of the BMBL that validation be performed as 
frequently as necessary at the discretion of the IBC and at least once 
annually to ensure that the centralized effluent heat treatment system 
is performing as intended under the established process parameters. 
This amendment shall apply to heat treatment systems used in both large 
animal BL3 and BL4 facilities.
    For large animal BL3 laboratories, the requirement for 
decontamination and inactivation (BL3-N) found at Appendix Q-II-C-1-b-
(5) currently states:

    Appendix Q-II-C-1-b-(5). Liquid effluent from containment 
equipment, sinks, biological safety cabinets, animal rooms, primary 
barriers, floor drains, and sterilizers shall be decontaminated by 
heat treatment before being released into the sanitary system. The 
procedure used for heat decontamination of liquid wastes shall be 
monitored with a recording thermometer. The effectiveness of the 
heat decontamination process system shall be revalidated every 30 
days with an indicator organism.

    Appendix Q-II-C-1-b-(5) is amended as follows:
    Appendix Q-II-C-1-b-(5). Liquid effluent from containment 
equipment, sinks, biological safety cabinets, animal rooms, primary 
barriers, floor drains, and sterilizers shall be decontaminated by heat 
treatment before being released into the sanitary system. The procedure 
used for heat decontamination of liquid wastes shall be monitored with 
a recording thermometer. The effectiveness of the heat decontamination 
process system shall be revalidated at minimum on a yearly basis with 
an indicator organism. More frequent validation, based on the amount of 
use or other safety factors, shall be left to the discretion of the 
IBC.
    For large animal BL3 laboratories, the requirement for animal 
facilities (BL3-N) found at Appendix Q-II-C-2-h currently states:

    Appendix Q-II-C-2-h. Liquid effluent from containment equipment, 
sinks, biological safety cabinets, animal rooms, primary barriers, 
floor drains, and sterilizers shall be decontaminated by heat 
treatment before being released into the sanitary system. The 
procedure used for heat decontamination of liquid wastes shall be 
monitored with a recording thermometer. The effectiveness of the 
heat decontamination process system shall be revalidated every 30 
days with an indicator organism.

    Appendix Q-II-C-2-h is amended as follows:
    Appendix Q-II-C-2-h. Liquid effluent from containment equipment, 
sinks, biological safety cabinets, animal rooms, primary barriers, 
floor drains, and sterilizers shall be decontaminated by heat treatment 
before being released into the sanitary system. The procedure used for 
heat decontamination of liquid wastes shall be monitored with a 
recording thermometer. The effectiveness of the heat decontamination 
process system shall be revalidated at minimum on a yearly basis with 
an indicator organism. More frequent validation, based on the amount of 
use or other safety factors, shall be left to the discretion of the 
IBC.
    For large animal BL4 laboratories, the requirement for 
decontamination and inactivation (BL4-N) found at Appendix Q-II-D-1-b-
(9) currently states:

    Appendix Q-II-D-1-b-(9). Liquid effluent from containment 
equipment, sinks, biological safety cabinets, animal rooms, primary 
barriers, floor drains, and sterilizers shall be decontaminated by 
heat treatment before being released into the sanitary

[[Page 22289]]

system. Liquid wastes from shower rooms and toilets shall be 
decontaminated with chemical disinfectants or heat by methods 
demonstrated to be effective. The procedure used for heat 
decontamination of liquid wastes shall be monitored with a recording 
thermometer. The effectiveness of the heat decontamination process 
system shall be revalidated every 30 days with an indicator 
organism. Liquid wastes from the shower shall be chemically 
decontaminated using an Environmental Protection Agency-approved 
germicide. The efficacy of the chemical treatment process shall be 
validated with an indicator organism. Chemical disinfectants shall 
be neutralized or diluted before release into general effluent waste 
systems.

    Appendix Q-II-D-1-b-(9) is amended as follows:
    Appendix Q-II-D-1-b-(9). Liquid effluent from containment 
equipment, sinks, biological safety cabinets, animal rooms, primary 
barriers, floor drains, and sterilizers shall be decontaminated by heat 
treatment before being released into the sanitary system. If required 
by design, regulation, local ordinance or policy, liquid wastes from 
shower rooms and toilets shall be decontaminated with chemical 
disinfectants or heat by methods demonstrated to be effective. The 
procedure used for heat decontamination of liquid wastes shall be 
monitored with a recording thermometer. The effectiveness of the heat 
decontamination process system shall be revalidated at minimum on a 
yearly basis with an indicator organism. More frequent validation, 
based on the amount of use or other safety factors, shall be left to 
the discretion of the IBC. If required by design, regulation, local 
ordinance or policy, liquid wastes from the shower shall be chemically 
decontaminated using an Environmental Protection Agency-approved 
germicide. The efficacy of the chemical treatment process shall be 
validated with an indicator organism. Chemical disinfectants shall be 
neutralized or diluted before release into general effluent waste 
systems.
    For large animal BL4 laboratories, the requirement for animal 
facilities (BL4-N) found at Appendix Q-II-D-2-i currently states:

    Appendix Q-II-D-2-i. Liquid effluent from containment equipment, 
sinks, biological safety cabinets, animal rooms, primary barriers, 
floor drains, and sterilizers shall be decontaminated by heat 
treatment before being released into the sanitary system. Liquid 
wastes from shower rooms and toilets shall be decontaminated with 
chemical disinfectants or heat by methods demonstrated to be 
effective. The procedure used for heat decontamination of liquid 
wastes shall be monitored with a recording thermometer. The 
effectiveness of the heat decontamination process system shall be 
revalidated every 30 days with an indicator organism. Liquid wastes 
from the shower shall be chemically decontaminated using an 
Environmental Protection Agency-approved germicide. The efficacy of 
the chemical treatment process shall be validated with an indicator 
organism. Chemical disinfectants shall be neutralized or diluted 
before release into general effluent waste systems.

    Appendix Q-II-D-2-i is amended as follows:
    Appendix Q-II-D-2-i. Liquid effluent from containment equipment, 
sinks, biological safety cabinets, animal rooms, primary barriers, 
floor drains, and sterilizers shall be decontaminated by heat treatment 
before being released into the sanitary system. If required by design, 
regulation, local ordinance or policy, liquid wastes from shower rooms 
and toilets shall be decontaminated with chemical disinfectants or heat 
by methods demonstrated to be effective. The procedure used for heat 
decontamination of liquid wastes shall be monitored with a recording 
thermometer. The effectiveness of the heat decontamination process 
system shall be revalidated at minimum on a yearly basis with an 
indicator organism. More frequent validation, based on the amount of 
use or other safety factors, shall be left to the discretion of the 
IBC. If required by design, regulation, local ordinance or policy, 
liquid wastes from the shower shall be chemically decontaminated using 
an Environmental Protection Agency-approved germicide. The efficacy of 
the chemical treatment process shall be validated with an indicator 
organism. Chemical disinfectants shall be neutralized or diluted before 
release into general effluent waste systems.

     Dated: April 9, 2016.
Lawrence A. Tabak,
Deputy Director, National Institutes of Health.
[FR Doc. 2016-08810 Filed 4-14-16; 8:45 am]
 BILLING CODE 4140-01-P