Document ID: EPA-HQ-OPPT-2011-0740-0018
Agency: epa
Document Type: Rule
Title: Microorganisms; General Exemptions from Reporting Requirements; Revisions to Recipient Organisms Eligible for Tier I and Tier II Exemptions
Posted Date: 2020-03-10T04:00Z

[Federal Register Volume 85, Number 47 (Tuesday, March 10, 2020)]
[Rules and Regulations]
[Pages 13760-13773]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2020-04746]

[[Page 13760]]

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ENVIRONMENTAL PROTECTION AGENCY

40 CFR Part 725

[EPA-HQ-OPPT-2011-0740; FRL-9991-60]
RIN 2070-AJ65

Microorganisms; General Exemptions From Reporting Requirements; 
Revisions to Recipient Organisms Eligible for Tier I and Tier II 
Exemptions

AGENCY: Environmental Protection Agency (EPA).

ACTION: Final rule.

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SUMMARY: EPA is issuing a final rule to add Trichoderma reesei (T. 
reesei) strain QM6a and its derivatives and Bacillus amyloliquefaciens 
(B. amyloliquefaciens) subspecies (subsp.) amyloliquefaciens to the 
list of recipient microorganisms that may be used to qualify for the 
Tier I and Tier II exemptions from full notification and reporting 
procedures under the Toxic Substances Control Act (TSCA) for new 
microorganisms that are being manufactured for introduction into 
commerce. EPA received petitions to add T. reesei and B. 
amyloliquefaciens to the list of microorganisms eligible for the 
exemption from full notification and reporting procedures under the 
TSCA for new microorganisms. Based on EPA's evaluation of these 
petitions, EPA has made the determination that certain strains of both 
microorganisms will not present an unreasonable risk of injury to 
health or the environment when used as a recipient microorganism 
provided that certain criteria for the introduced genetic material and 
the physical containment conditions are met.

DATES: This final rule is effective April 9, 2020.

ADDRESSES: The docket for this action, identified by docket 
identification (ID) number EPA-HQ-OPPT-2011-0740, is available at 
http://www.regulations.gov or at the Office of Pollution Prevention and 
Toxics Docket (OPPT Docket), Environmental Protection Agency Docket 
Center (EPA/DC), West William Jefferson Clinton Bldg., Rm. 3334, 1301 
Constitution Ave. NW, Washington, DC. The Public Reading Room is open 
from 8:30 a.m. to 4:30 p.m., Monday through Friday, excluding legal 
holidays. The telephone number for the Public Reading Room is (202) 
566-1744, and the telephone number for the OPPT Docket is (202) 566-
0280. Please review the visitor instructions and additional information 
about the docket available at http://www.epa.gov/dockets.

FOR FURTHER INFORMATION CONTACT: 
    For technical information contact: Rebecca Edelstein, Chemical 
Control Division (7405M), Office of Pollution Prevention and Toxics, 
Environmental Protection Agency, 1200 Pennsylvania Ave. NW, Washington, 
DC 20460-0001; telephone number: (202) 564-1667; email address: 
edelstein.rebecca@epa.gov.
    For general information contact: The TSCA-Hotline, ABVI-Goodwill, 
422 South Clinton Ave., Rochester, NY 14620; telephone number: (202) 
554-1404; email address: TSCA-Hotline@epa.gov.

SUPPLEMENTARY INFORMATION:

I. Executive Summary

A. Does this action apply to me?

    You may be potentially affected by this action if you produce, 
import, process, or use either intergeneric T. reesei or intergeneric 
B. amyloliquefaciens or any other eligible recipient microorganisms 
listed in 40 CFR 725.420. The following list of North American 
Industrial Classification System (NAICS) codes is not intended to be 
exhaustive, but rather provides a guide to help readers determine 
whether this document applies to them. Potentially affected entities 
may include:
     Basic Chemical Manufacturing (NAICS code 3251).
     Pesticide, Fertilizer and other Agricultural Chemical 
manufacturing (NAICS code 3253).
     Other Chemical Product and Preparation Manufacturing 
(NAICS code 3259).
    If you have any questions regarding the applicability of this 
action to a particular entity, consult the technical person listed 
under FOR FURTHER INFORMATION CONTACT.

B. What is the Agency's authority for taking this action?

    This action is being taken under the authority of TSCA section 
5(h)(4) (15 U.S.C. 2604(h)(4)). TSCA section 5(a)(1) requires that 
persons notify EPA at least 90 days before they manufacture (the term 
``manufacture'' includes import under TSCA) for commercial purposes a 
``new'' chemical substance, or manufacture (including import) or 
process a chemical substance for a ``significant new use'' (15 U.S.C. 
2604(a)(1)(B)(i)). TSCA furthermore prohibits such manufacturing or 
processing from commencing until EPA has conducted a review of the 
notice, made an appropriate determination on the notice, and taken such 
actions as are required in association with that determination (15 
U.S.C. 2604(a)(1)(B)(ii)). TSCA defines ``chemical substance'' broadly 
and in terms that cover intergeneric microorganisms as well as 
traditional chemical substances. Therefore, for the purposes of TSCA, a 
``new microorganism'' is one that is not listed on the TSCA Chemical 
Substances Inventory (TSCA Inventory) compiled under TSCA section 8(b).
    TSCA section 5(h)(4) authorizes EPA, upon application and by rule, 
to exempt the manufacturer of any new chemical substance from part or 
all of the provisions of TSCA section 5, if EPA determines that the 
manufacture, processing, distribution in commerce, use, or disposal of 
the new chemical substance will not present an unreasonable risk of 
injury to human health or the environment, including an unreasonable 
risk to a potentially exposed or susceptible subpopulation identified 
by the Administrator under the conditions of use.

C. What action is the Agency taking?

    In 2012, EPA proposed to add T. reesei strain QM6a and its 
derivatives (hereafter, T. reesei QM6a) and B. amyloliquefaciens 
subspecies (subsp.) amyloliquefaciens to the list of recipient 
microorganisms in 40 CFR 725.420 that may be used to qualify for Tier I 
and Tier II exemptions from full notification and reporting procedures 
under TSCA for new microorganisms that are being manufactured into 
commerce. EPA is finalizing the proposal.

D. Why is the Agency taking this action?

    EPA received petitions to add T. reesei and B. amyloliquefaciens to 
the list of microorganisms that may be used as recipient microorganisms 
in order to qualify for the exemption from full notification and 
reporting procedures under TSCA for new microorganisms that are being 
manufactured for introduction into commerce. EPA proposed to add 
certain strains of these two microorganisms to the list of recipient 
microorganisms based on EPA's preliminary determination. EPA is now 
issuing a final rule that incorporates certain changes in response to 
public comment.

E. What are the estimated incremental impacts of this final rule?

    EPA has evaluated the potential costs of the addition of the two 
microorganisms to the list of recipient microorganisms eligible for 
Tier I and Tier II exemptions. The final rule is

[[Page 13761]]

expected to generate cost savings for organizations that, in the 
absence of the rule, would submit Microbial Commercial Activity Notices 
(MCANs) for new intergeneric T. reesei or B. amyloliquefaciens strains. 
The rule will result in costs savings for both the industry and the 
Agency. EPA estimates the annualized industry savings of the rule to be 
approximately $260,000 per year over a ten-year period, with a 3 
percent discount rate, and $252,000 per year with a 7 percent discount 
rate. Annualized agency savings are approximately $178,000 per year 
with a 3 percent discount rate and $173,000 per year with a 7 percent 
discount rate over the ten-year period, for a total annualized savings 
to society of approximately $438,000 per year with a 3 percent discount 
rate and $424,000 per year with a 7 percent discount rate. The economic 
analysis is available in the docket and is summarized in Unit IX. of 
this final rule. Costs and benefits of adding T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens to 40 CFR 725.420 are also 
discussed in Unit VIII.C.2. through 4.

II. Background

    EPA received petitions to add T. reesei and B. amyloliquefaciens to 
the list of recipient microorganisms at 40 CFR 725.420 that are 
eligible for the regulatory exemptions applicable to new microorganisms 
that are manufactured for introduction into commerce (Refs. 1-3). In 
the Federal Register of September 5, 2012 (77 FR 54499) (FRL-9348-1) 
(``2012 Proposed Rule'') (Ref. 4), the Agency proposed to add certain 
strains of these two microorganisms to the list of recipient 
microorganisms at 40 CFR 725.420 based on EPA's preliminary 
determination that both of the microorganisms, with certain 
limitations, meet the criteria for addition to the list--i.e., they 
will not present an unreasonable risk of injury to health or the 
environment provided that the other conditions of the exemptions at 40 
CFR part 725, subpart G, relating to the introduced genetic material, 
and the physical containment of the new microorganisms, have been met. 
EPA is now issuing a final rule that incorporates certain changes made 
in response to public comments received on the 2012 Proposed Rule. 
These changes are described in the following paragraphs.
    In the 2012 Proposed Rule, EPA proposed to restrict the exemption 
for T. reesei to the T. reesei strain QM6a and its derivatives. In 
addition, EPA proposed to restrict the T. reesei QM6a exemption to use 
of the microorganism only under submerged standard industrial 
fermentation operations used for enzyme production; as described in 
this proposed rule, these conditions are typical throughout the 
fermentation industry and meet the existing physical containment and 
control requirements for the tiered exemptions under 40 CFR 725.422. 
Any subsequent deliberate fermentation of solid plant material or 
insoluble substrates with T. reesei QM6a and its derivatives as defined 
at 40 CFR 725.3 could only be initiated after inactivation of the 
viable T. reesei cells as delineated in 40 CFR 725.422(d), i.e., by a 
procedure that has been demonstrated and documented to be effective in 
reducing the viable microbial population by at least 6 logs (i.e., six 
orders of magnitude).
    In addition, EPA proposed to limit the exemption for B. 
amyloliquefaciens to only strains of B. amyloliquefaciens that would 
fall under the subspecies B. amyloliquefaciens amyloliquefaciens.
    In response to comments received on its original proposal, EPA has 
modified the regulatory text in 40 CFR 725.3 and 725.420 slightly to 
better clarify EPA's original intent. These revisions to the regulatory 
text in 40 CFR 725.3 and 725.420 merely represent a clarification of 
the original proposal.
    Existing regulatory requirements and exemptions for intergeneric 
microorganisms are discussed in Unit III. of this proposed rule. EPA's 
response to public comments received on the 2012 Proposed Rule are 
provided in Unit IV. Unit V. provides EPA's evaluation of available 
information on T. reesei and B. amyloliquefaciens for the criteria 
delineated in 40 CFR 725.67. Physical containment and control 
technologies as well as release and exposure assessments for the two 
microorganisms are discussed in Unit VI. EPA's risk assessments for T. 
reesei QM6a and B. amyloliquefaciens subsp. amyloliquefaciens are 
summarized in Unit VII., and EPA's rationale for adding the two 
microorganisms to the list of recipients eligible for exemption is 
discussed in Unit VIII. EPA's Risk Assessment documents (Refs. 5 and 
6), available in the public docket, provide more detailed information, 
and supporting references, for EPA's evaluation of the available 
information and the potential risks to health and the environment.

III. Existing EPA Regulatory Requirements and Exemption Standard

    Manufacturers are required to report certain information to EPA 90 
days before commencing the manufacture of intergeneric microorganisms 
that are not listed on the TSCA Inventory. EPA regulations at 40 CFR 
part 725 establish the mechanisms for reporting this information. TSCA 
prohibits such manufacturing or processing from commencing until EPA 
has conducted a review of the notice, made an appropriate determination 
on the notice, and taken such actions as are required in association 
with that determination (15 U.S.C. 2604(a)(1)(B)(ii)).
    Any manufacturer of a living intergeneric microorganism who is 
required to report under TSCA section 5 must file a MCAN with EPA, 
unless the activity is eligible for one of the specific exemptions. 
Section 5(h)(4) authorizes EPA, by rule and upon request, to exempt 
manufacturers from these requirements if the Administrator determines 
that the manufacture, processing, distribution in commerce, use or 
disposal of the chemical substance ``will not present an unreasonable 
risk of injury to health or the environment, including an unreasonable 
risk to a potentially exposed or susceptible subpopulation identified 
by the Administrator under the conditions of use.'' TSCA section 3(4) 
defines ``conditions of use'' to mean ``the circumstances, as 
determined by the Administrator, under which a chemical substance is 
intended, known or reasonably foreseen to be manufactured, processed, 
distributed in commerce, used, or disposed of.'' TSCA section 3(12) 
defines ``potentially exposed or susceptible subpopulation'' to mean 
``a group of individuals within the general population. . .who, due to 
either greater susceptibility or greater exposure, may be at greater 
risk than the general population of adverse health effects from 
exposure to a chemical substance or mixture, such as infants, children, 
pregnant women, workers, or the elderly.'' 15 U.S.C. 2601 et seq. The 
general procedures for filing MCANs are described in 40 CFR part 725, 
subpart B.
    EPA regulations establish two exemptions for new microorganisms, 
after the research and development stage, which are being manufactured 
for introduction into commerce: Tier I and Tier II exemptions.
    Under the Tier I exemption, if certain criteria are met, 
manufacturers are required to notify EPA 10 days prior to manufacturing 
a new microorganism that qualifies for this exemption, and to keep 
certain records. 40 CFR 725.400. To qualify for the Tier I exemption, a 
manufacturer must use one of the recipient organisms listed in 40 CFR 
725.420, and must implement specific physical containment and control 
technologies listed in 40 CFR 725.422. In addition, the genetic 
material

[[Page 13762]]

introduced into the recipient microorganism must be well-characterized, 
limited in size, poorly mobilizable, and free of certain sequences. 40 
CFR 725.421.
    A manufacturer who meets the conditions of the Tier I exemption may 
modify the specified containment restrictions or level of inactivation 
but must submit a Tier II exemption notification 40 CFR 725.428. The 
Tier II exemption requires manufacturers to submit an abbreviated 
notification describing the modified containment and provides for a 45-
day period during which EPA would review the proposed containment. 40 
CFR 725.450 and 725.470. The manufacturer may not proceed under this 
exemption until EPA approves the exemption. 40 CFR 725.470.
    EPA established a petition process at 40 CFR 725.67 for the public 
to propose additional microorganisms for the tiered exemptions. EPA's 
regulations at 40 CFR 725.67 direct petitioners to submit information 
to demonstrate that the activities affected by the requested exemption 
meet the requirements of TSCA section 5(h)(4), i.e. ``will not present 
an unreasonable risk of injury to health or the environment, including 
an unreasonable risk to a potentially exposed or susceptible 
subpopulation identified by the Administrator under the conditions of 
use.'' 15 U.S.C. 2604(h)(4). In addition, a petitioner is responsible 
for providing supporting information for this determination in four 
general categories:

    1. The effects of the new microorganism on health and the 
environment.
    2. The magnitude of exposure of human beings and the environment 
to the new microorganism.
    3. The benefits of the new microorganism for various uses and 
the availability of substitutes for such uses.
    4. The reasonably ascertainable economic consequences of 
granting or denying the petition, including effects on the national 
economy, small business, and technological innovation.

    The regulations at 40 CFR 725.67 specify that when applying to list 
a recipient microorganism for the tiered exemption under 40 CFR 
725.420, petitioners should include information addressing six 
specified criteria, which EPA will use to evaluate the microorganism 
for listing. 40 CFR 725.67(a)(3)(iii). The six criteria are:

    1. Identification and classification of the microorganism using 
available genotypic and phenotypic information.
    2. Information to evaluate the relationship of the microorganism 
to any other closely related microorganisms which have a potential 
for adverse effects on health or the environment.
    3. A history of safe commercial use for the microorganism.
    4. Commercial uses indicating that the microorganism products 
might be subject to TSCA.
    5. Studies which indicate the potential for the microorganism to 
cause adverse effects to health or the environment.
    6. Studies which indicate the survival characteristics of the 
microorganism in the environment.

IV. Response to Public Comments on the 2012 Proposed Rule

    The Agency received three comments on the 2012 Proposed Rule (Ref. 
4). One comment, from an anonymous submitter (Ref. 7), concerned mold 
problems in rental housing and thus was not relevant to the proposed 
rule. A second comment, from an individual (Ref. 8), supported the 
proposed rule.
    The third comment was a joint set of comments from the 
Biotechnology Industry Organization (BIO) and the Enzyme Technical 
Association (ETA) (Ref. 9). While generally supportive of the proposed 
rule, BIO/ETA raised three important issues with respect to EPA's 
proposed rule.
    First, BIO/ETA expressed concern that the proposed wording in 
section 725.420(k), that reads ``Trichoderma reesei strain QM6a used 
only in . . .'' does not accurately reflect the range of T. reesei 
strain QM6a microorganisms currently being used in standard industrial 
fermentations. BIO/ETA requested that the phrase be reworded as 
``Trichoderma reesei strain QM6a and its derivatives used only in . . 
.'' EPA agrees that the commenter's suggested language more accurately 
reflects the Agency's original intent. EPA did not originally intend to 
restrict the exemption to the naturally occurring QM6a isolate. Most of 
the strains of T. reesei currently used in industrial production are 
not the naturally occurring QM6a isolate, but are strains derived from 
QM6a that have been modified by physical or chemical mutagenesis to 
obtain microorganisms with improved enzyme-producing abilities. 
Accordingly, EPA has adopted the commenter's suggested revision to 
clarify that the exemption applies not only to the naturally occurring 
strain, but also to any strain derived from the naturally occurring 
QM6a.
    Second, BIO/ETA expressed concern that the proposed regulation was 
too broadly worded and as drafted would not clearly distinguish between 
standard industrial fermentation operations used to produce enzymes, 
and fermentation operations conducted for other purposes. Specifically, 
the commenter raised concern that the inclusion of an unqualified 
restriction in proposed 40 CFR 725.420(k) that ``no solid plant 
material or insoluble substrate is present in the fermentation broth'' 
would prohibit the use of T. reesei in submerged standard industrial 
fermentation operations used for enzyme production. Enzyme production 
is the first phase of some industrial applications such as cellulosic 
ethanol production where the first fermentation is to grow the 
microorganism to produce enzymes, followed by another fermentation of 
pretreated plant biomass for conversion of the cellulose and 
hemicellulose to simple sugars (i.e., saccharification), followed by a 
third fermentation of the sugars to ethanol by yeast or another 
ethanologen. As part of the process of growing the microbes for enzyme 
production by T. reesei QM6a and its derivatives, nutrients need to be 
available, including those from plant materials such as soy or corn, 
which may contain insoluble components. The second fermentation 
operation of saccharification of plant biomass may occur only after the 
T. reesei microorganism has been inactivated. The use of nutrients 
supplied by plant material (e.g., soy meal, corn steep liquor) in the 
first fermentation for enzyme production has a long history of safe 
use.
    To address this issue, the commenter suggested revising the 
regulatory text to ensure that the typical industry practice of 
supplying nutrients in the form of solid plant materials during the 
initial enzyme fermentation would fall within the scope of the proposed 
exemption. EPA agrees and is therefore changing the regulatory text to 
allow the use of solid plant material in the enzyme fermentation step. 
Under the final regulatory text, the use of the conventional 
fermentation ingredients from solid plant material--for example, soy or 
corn meal and other insoluble fermentation ingredients from corn or soy 
which contain insoluble components--is allowed when used specifically 
to provide nutrients for growth of the microorganism during standard 
enzyme fermentation as described in part 1 of the definition at 40 CFR 
725.3.
    The commenter further suggested adding text to clarify that the 
requirement to inactivate the organism applies prior to ``subsequent 
fermentation operations, and not to the initial enzyme production 
stage.'' EPA agrees that the commenter has identified a reasonable 
basis for concern with respect to the proposed regulatory text. EPA 
acknowledges that nutrients for microbial growth in submerged standard 
industrial fermentation during the initial enzyme production phase of 
the

[[Page 13763]]

fermentation operation may be supplied by soybean meal, corn steep 
liquor, or other plant-derived materials that may contain insoluble 
substrates. The use of such plant materials as nutrient sources for 
microbial growth in submerged standard industrial fermentation 
operations used for enzyme production is a standard industry practice 
with a long history of safe use, and it does not result in the 
production of secondary toxic metabolites such as paracelsin because 
the fermentations involve the logarithmic growth of the cells in the 
presence of optimal concentrations of carbon and nitrogen and other 
nutrients (see Unit V. for more detail on paracelsin). EPA did not 
originally intend to preclude such operations and agrees that revision 
to the regulatory text is warranted to clarify that solid plant 
material can be used to provide nutrients for growth of the 
microorganism during submerged standard enzyme fermentation operations.
    However, EPA continues to have concern about the potential for the 
production of paracelsin during the second fermentation phase of 
cellulosic ethanol production, i.e., the saccharification of the 
pretreated plant biomass, because of the presence of solid surfaces and 
an excess of carbon substrate with live T. reesei QM6a (and its 
derivatives) cells. Therefore, EPA is retaining the requirement that 
fermentation operations subsequent to the enzyme production 
fermentation phase may only be initiated after inactivation of the 
viable T. reesei cells as delineated in 40 CFR 725.422(d) (i.e., by a 
procedure that has been demonstrated and documented to be effective in 
reducing the viable microbial population by at least 6 logs). 
Inactivation of T. reesei QM6a prior to a subsequent or secondary 
fermentation that may contain solid plant material or insoluble 
substrates (as defined at 40 CFR 725.3) avoids the potential for 
production of paracelsin.
    BIO/ETA also commented that paracelsin may be produced under non-
standard conditions of fermentation, such as ``surface fermentation 
media with large concentrations of biomass,'' and requested that EPA 
revise the rule to reflect this. EPA interprets this comment to mean 
``surface fermentation media with large concentrations of biomass'' is 
the only condition under which paracelsin can be produced and that BIO/
ETA is requesting that the rule be amended accordingly. EPA agrees that 
paracelsin may be produced under non-standard conditions of 
fermentation, such as surface fermentation with large concentrations of 
biomass. However, available scientific literature indicates that 
paracelsin may also be produced under certain other fermentation 
conditions. Scientific literature suggests that surface fermentation is 
synonymous with solid-state fermentation where microorganisms are grown 
on the surface of a solid support that is not submerged. While it is 
likely that the potential for paracelsin production is greater with 
solid-state/surface fermentation, the production of peptaibols (of 
which paracelsin is one) by Trichoderma species has been shown to occur 
even in liquid broth culture in the presence of plant material or 
insoluble substrates in laboratory studies. Thus, paracelsin production 
potentially may be produced in fermentation broth amended with plant 
material providing excess carbon. Therefore, EPA is not amending the 
rule to indicate that the only conditions in which paracelsin 
potentially may be produced are with surface fermentations with large 
concentrations of biomass.

V. EPA's Evaluation of Available Information on T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens for the Criteria Delineated

    Pursuant to 40 CFR 725.67, Genencor International, Inc., 
(subsequently supported by the Enzyme Technical Association (ETA)) and 
Novozymes North America, Inc., submitted Letters of Application to EPA 
requesting that T. reesei and B. amyloliquefaciens (Refs. 1 and 2) be 
added to 40 CFR 725.420 as candidate recipient microorganisms for the 
tiered exemptions. The letters of application provided information that 
the submitters believed demonstrate that activities affected by the 
requested exemptions would not present an unreasonable risk of injury 
to health or the environment. Information regarding the criteria 
specified in 40 CFR 725.67(a)(2) and 725.67(a)(3)(iii) were addressed 
in these letters of application to list T. reesei and B. 
amyloliquefaciens as recipient microorganisms under 40 CFR 725.420.
    EPA has made the determination based on the information provided in 
the Letters of Application (Refs. 1 and 2), supplemental information 
provided by ETA (Refs. 10 and 11), and other information available to 
EPA that T. reesei QM6a, with certain restrictions, and B. 
amyloliquefaciens subsp. amyloliquefaciens will not present an 
unreasonable risk of injury to health or the environment, including an 
unreasonable risk to a potentially exposed or susceptible 
subpopulation, when used as recipient microorganisms provided that: (a) 
The existing criteria for the introduced genetic material listed in 40 
CFR 725.422 are met, and (b) the physical containment and control 
technologies criteria listed at 40 CFR 725.422 are met. In making this 
determination, EPA identified workers as a potentially exposed or 
susceptible subpopulation to the substances under the conditions of use 
and concluded that, with the limitations described above, the 
substances will not present an unreasonable risk of injury to health or 
the environment. EPA's Risk Assessments for these two microorganisms 
(Refs. 5 and 6) are available in the docket.
    This unit presents a summary of EPA's evaluation of the available 
information pertinent to the six criteria delineated in 40 CFR 
725.67(a)(3)(iii) for both microorganisms.

A. Evaluation of Available Information Relevant to the Criteria for T. 
reesei QM6a as a Recipient Microorganism with Specified Conditions of 
Growth

    1. Identification and classification of the microorganism using 
available genotypic and phenotypic information.
    T. reesei is a hypercellulolytic fungus originally isolated in the 
Solomon Islands in 1944. T. reesei was found on deteriorating military 
fabrics such as tents and clothing. This isolate, designated as QM6a, 
was initially named Trichoderma viride. Approximately 20 years later, 
QM6a was re-classified as Trichoderma reesei.
    T. reesei is the species name given to the anamorphic form (this 
form reproduces asexually) of the fungus whose teleomorphic form (this 
form reproduces sexually) is now understood to be Hypocrea jecorina.
    Recent taxonomic studies have shown that the species T. reesei 
consists only of this single isolate QM6a and its derivatives. Many 
other strains called T. reesei isolated elsewhere have now been 
proposed as belonging to a newly named species, T. parareesei, based on 
differences in habitat, sporulation, and metabolic versatility. T. 
reesei has been shown to belong to a single species now referred to as 
H. jecorina/T. reesei (QM6a) which reflects its relationship to its 
teleomorph H. jecorina. The only anamorphic strains within the species 
H. jecorina/T. reesei are those of QM6a and its derivatives. The 
petition to add T. reesei to the list of microorganisms at 40 CFR 
725.420 requested that EPA include all strains of T. reesei. However, 
given these recent taxonomic publications, all fungal strains correctly 
named T. reesei are, by definition, QM6a or a derivative.

[[Page 13764]]

    Adequate genotypic and phenotypic information is available for 
classification of T. reesei QM6a and its derivatives. The American Type 
Culture Collection (ATCC) designation for this original strain of T. 
reesei QM6a is ATCC 13631.
    2. Information to evaluate the relationship of the microorganism to 
any other closely related microorganisms that have a potential for 
adverse effects on health or the environment.
    Closely related members of section Longibrachiatum do not have a 
potential for adverse effects; other less closely related Trichoderma 
species have a potential to cause adverse effects as pathogens of 
commercially produced mushrooms. These less closely related species 
include various species of the Harzianum clade, T. aggressivum, T. 
pleuotrophilum, and T. fulvidum that are responsible for significant 
loss of the mushroom crops of Agaricus bisporus and Pleurotus 
ostreatus.
    T. reesei/H. jecorina can be distinguished from other Trichoderma 
species by a comprehensive approach employing criteria of the 
Genealogical Concordance Phylogenetic Species Recognition (GCPSR) 
concept, which commonly requires the use of genealogies of three or 
four genes, not just the sequences of spacer regions as previously 
utilized for identification. Use of the GCPSR protocol would separate 
T. reesei (sensu lato) from the opportunistic pathogens within the 
section Longibrachiatum, including T. longibrachiatum and T. 
citrinoviride/H. schweinitzii, as well as the mold disease pathogens of 
mushrooms.
    3. A history of safe commercial use for the microorganism.
    T. reesei QM6a has a long history of safe use producing a variety 
of commercial enzymes. T. reesei QM6a cellulases, beta-glucanases, and 
xylanases are used by the animal feed, baking, beverages, textile 
processing, detergent, pulp and paper, industrial chemicals, and 
biofuels industries.
    For industrial enzyme production, T. reesei is generally grown in a 
closed, submerged standard industrial fermentation system. In submerged 
standard industrial fermentation operations used for enzyme production, 
growth of the microorganism occurs beneath the surface of the liquid 
growth medium. As described in this unit, this type of fermentation 
system appears to be typical throughout the industry, based on EPA's 
review of MCAN submissions over the years.
    Under this type of fermentation system, the fermentation broth is a 
defined mixture of carbon and nitrogen sources, some of which may be 
supplied from plant material or soluble substrates (e.g., soy meal, 
corn steep liquor), minerals, salts, and other nutrients, is maintained 
at optimal pH and temperature, and is typically aerated and mixed. 
These conditions support the active growth and productivity of the 
organisms for enzyme production. Submerged standard industrial 
fermentation operations used for enzyme production systems reduce the 
potential for exposure of workers to the production organism and 
fermentation broth aerosols, reduce the potential for contamination of 
the culture and make the collection of extracellular enzymes simpler 
and less costly. The fermentation process is terminated before the T. 
reesei QM6a organisms go into the stationary growth phase (i.e., before 
secondary metabolism begins). At the end of the fermentation process, 
the production organisms are separated from the fermentation broth and 
inactivated.
    Several enzymes produced by T. reesei QM6a have Generally 
Recognized as Safe (GRAS) status with the Food and Drug Administration 
(FDA) or FDA had no questions about the GRAS conclusions about them 
contained in GRAS submissions to FDA. This supports the Agency's 
conclusion that commercial use of T. reesei QM6a and its derivatives as 
a recipient microorganism for commercial enzyme production will not 
present an unreasonable risk of injury to health or the environment. T. 
reesei QM6a enzymes used in foods that have been granted GRAS status or 
for whose claimed GRAS status FDA had no questions include cellulase, 
hemicellulase, transglucosidase, pectin lyase, acid fungal protease, 
and a chymosin enzyme preparation. Data supporting the GRAS notices 
included the results of pathogenicity tests for the T. reesei QM6a 
production organisms and toxicity tests for the enzyme products. The 
data showed that the production strains are not pathogenic and did not 
produce toxins during enzyme fermentation.
    4. Commercial uses indicating that the microorganism products might 
be subject to TSCA.
    EPA has reviewed 48 MCANs involving intergeneric T. reesei 
production organisms used to manufacture a number of industrial 
enzymes, including amylases, glucosidases, proteases, phytase, laccase, 
and numerous cellulolytic enzyme preparations. Amylases and 
glucosidases are used for the breakdown of starch into sugars and have 
been used in laundry detergents and in textile processing. More 
recently, industrial enzymes produced by T. reesei have been produced 
for corn and cellulosic ethanol production. T. reesei produces numerous 
cellulases and hemicellulases that are efficient in degrading plant 
biomass. Intergeneric T. reesei strains could also be used to 
manufacture industrial chemicals other than enzymes such as surfactants 
or specialty chemicals. More detailed information on MCANs submitted to 
EPA can be viewed on EPA's TSCA Biotechnology Program web page: https://www.epa.gov/regulation-biotechnology-under-tsca-and-fifra/overview-biotechnology-under-tsca.
    5. Studies which indicate the potential for the microorganism to 
cause adverse effects to health or the environment.
    a. Human health hazards -- i. Pathogenicity. T. reesei QM6a is not 
pathogenic to humans. Due to its long history of use for production of 
enzymes used in food applications, the potential for the fungus and its 
products to be pathogenic or toxic to humans has been evaluated 
numerous times. Various studies have been conducted assessing T. reesei 
QM6a's pathogenic potential in healthy and immunocompromised laboratory 
animals. With the exception of one study where a high inoculum of 
intravenous (iv) and intraperitoneal (ip) injection of spores in 
immunocompromised mice resulted in pathogenic effects, studies have 
demonstrated a lack of pathogenicity of T. reesei QM6a. Numerous 
pathogenicity studies have been conducted as part of GRAS notices to 
FDA for several different enzymes used in the food industry. Studies 
using injection of T. reesei QM6a in rats, using both healthy and 
immunosuppressed rats, and using ip injection of viable and heat-killed 
cells of T. reesei QM6a in rats have all demonstrated a lack of 
potential pathogenicity to humans.
    T. reesei QM6A is not known to possess any virulence factors 
associated with colonization or disease such as adherence factors, 
penetration factors, necrotic factors, toxins, or the ability to grow 
at human body temperature, 37 [deg]C. There are no reports of harmful 
effects associated with the use of or exposure to T. reesei QM6A 
strains, even after decades of commercial use for enzyme production. 
The body of evidence indicates that T. reesei QM6A does not pose 
concerns regarding human pathogenicity.
    ii. Toxicity. Available data indicate that T. reesei QM6a strains 
used in submerged standard industrial fermentation operations used for 
enzyme production do not present

[[Page 13765]]

human toxicity concerns. A number of studies have been conducted 
assessing the potential for T. reesei QM6a to produce toxins during 
submerged standard industrial fermentation operations used for enzyme 
production for food, pharmaceutical, or industrial uses. A cellulase 
enzyme known as celluclast produced by T. reesei QM6a has been tested 
for general oral toxicity and inhalation toxicity. Acute oral toxicity 
studies conducted in mice, rats, and dogs showed that T. reesei QM6a 
cellulase was not toxic to any of the test animals. Subchronic toxicity 
studies showed no evidence of systemic effects in dogs or rats. 
Additional toxicity studies have been conducted on other enzymes 
produced by T. reesei QM6a, the results of which have been presented in 
various GRAS petitions. Acute oral toxicity tests on two endoglucanases 
and a glucoamylase showed a lack of toxins. Subchronic feeding studies 
conducted on a cellulase, two xylanases, two endoglucanases, a 
protease, and a glucoamylase also showed a lack of toxicity in rats.
    Under typical industry practice, industrial fermentations of T. 
reesei QM6a for enzymes to be used in food are routinely checked by the 
enzyme producers to confirm the absence of antibiotic activity and 
toxins (Ref. 12). Relying on the data that show T. reesei QM6a has a 
long history of safe use in the production of food enzymes, EPA has 
concluded that strains used industrially would not be expected to 
produce these toxins under the conditions of submerged standard 
industrial fermentation used for enzyme production.
    iii. Mycotoxins and other secondary metabolites. The only health 
concern associated with T. reesei QM6a is its ability to produce a 
peptaibol secondary metabolite called paracelsin. Peptaibols are small 
linear peptides of 1,000-2,000 daltons characterized by a high content 
of the non-proteinogenic amino acid [alpha]-amino-isobutyric acid 
(Aib), with an N-terminus that is typically acetylated, and a C-
terminus that is linked to an amino alcohol, which is usually 
phenylalaninol, or sometimes valinol, leucinol, isoleucinol, or 
tryptophanol. Peptaibols are associated with a wide variety of 
biological activities and have antifungal, antibacterial, sometimes 
antiviral, antiparasitic, and neurotoxic activity. Paracelsin has been 
shown to damage mammalian cells such as human erythrocytes with an in 
vitro hemolytic activity of C50 = 3.7 x 10\-5\ mole/liter 
(mol/L) (Ref. 5).
    Paracelsin has not been detected in the use of T. reesei QM6a under 
the submerged standard industrial fermentation operations used for 
enzyme production, and numerous toxicity studies on enzyme products of 
T. reesei QM6a have demonstrated a lack of toxicity to laboratory 
animals. EPA therefore expects that paracelsin production would be of 
insignificant concern, provided the microorganisms are produced with 
submerged standard industrial fermentation operations used for enzyme 
production as described at 40 CFR 725.3.
    Under other conditions of fermentation, for example with the 
deliberate fermentation of cellulosic biomass for saccharification of 
plant material or extended fermentation, paracelsin may be produced 
(Ref. 5). Neither the information submitted with the petition, nor the 
information that is otherwise available is sufficient to allow EPA to 
determine the extent of paracelsin formation under these non-standard 
conditions. Consequently, EPA is unable to determine whether the use of 
the microbe under conditions other than submerged standard industrial 
fermentation operations used for enzyme production (i.e., specific 
conditions under which paracelsin is not expected to be formed) will 
not pose an unreasonable risk to human health and/or the environment 
(Ref. 5).
    b. Environmental hazards--i. Hazards to animals. T. reesei QM6a is 
not pathogenic to domesticated animals or wildlife. However, the 
secondary metabolite paracelsin has been shown to exhibit toxicity to 
aquatic species. A 24-hr exposure of paracelsin to Artemia salina 
(brine shrimp) resulted in a lethal concentration of 50% 
(LC50) of 21.26 micromoles ([micro]M) (40.84 micrograms per 
milliliter ([micro]g/ml)) which decreased to 9.66 [micro]M (18.56 
[micro]g/ml) with a 36-hr exposure. With Daphnia magna, paracelsin was 
found to be moderately toxic, with an LC50 of 7.70 [micro]M 
(14.79 [micro]g/ml) with a 24-hr exposure, and 5.60 [micro]M (10.76 
[micro]g/ml) with a 36-hr exposure.
    ii. Hazards to plants. T. reesei QM6a is not a pathogen of plants. 
Although it is capable of degrading cellulose and hemicellulose due to 
the copious quantities of the enzymes it can produce, it cannot be a 
primary colonizer on plant tissue. Genetic studies have shown that T. 
reesei QM6a does not contain any genes for ligninases, required for 
initial breakdown of plant material. This species is known as a wood 
rot fungus, but it apparently attacks only decaying plant material, not 
live plants.
    iii. Effects on other organisms. Peptaibols are toxic to Gram-
positive bacteria and various fungi. The inhibitory action of 
peptaibols on various fungi is the reason that many species of 
Trichoderma are used as biocontrol agents of plant pathogenic fungi. 
The peptaibol produced by T. reesei, QM6a paracelsin, has been shown to 
be inhibitory to one particular fungus, Phoma destructiva.
    Some species of Trichoderma, specifically T. aggressivum, T. 
pleuotrophilum, and T. fulvidum are pathogens of mushrooms. However, T. 
reesei QM6a is not a pathogen of mushrooms.
    6. Studies which indicate the survival characteristics of the 
microorganism in the environment. The species T. reesei is known only 
from the single original isolate QM6a from the Solomon Islands. 
Therefore, there is little information on its prevalence or behavior in 
the environment. Microcosm studies have been conducted that suggest it 
would survive in the environment in the plant rhizosphere and in bulk 
soils if inadvertently released.
    Although T. reesei was originally isolated from a tropical climatic 
region, it would be expected to persist in soils for extended periods 
of time, even after cold temperatures.

B. Evaluation of Available Information Relevant to the Criteria for B. 
amyloliquefaciens as a Recipient Microorganism

    1. Identification and classification of the microorganism using 
available genotypic and phenotypic information.
    B. amyloliquefaciens was initially proposed as a unique species in 
1943. The name B. amyloliquefaciens lost standing when it was not 
included on the Approved List of Bacterial Names with Standing in 
Nomenclature in 1980. Since classical phenotypic tests could not 
differentiate it as a unique species from Bacillus subtilis, it was 
regarded as a subspecies of B. subtilis for several decades. However, 
molecular evidence from subsequent studies led to the conclusion that 
B. amyloliquefaciens did indeed deserve independent status. The DNA 
homology between B. subtilis and B. amyloliquefaciens is only about 
15%. In addition, there were several phenotypic properties that 
differed between the two species. Chemotaxonomic studies revealed 
additional capability of separating strains of B. amyloliquefaciens 
from the other related species, Bacillus subtilis, Bacillus 
licheniformis, and Bacillus pumilus. The species has remained within 
the genus Bacillus sensu stricto since it was last established as a 
separate species.
    Recently, it has been proposed that there are two subspecies within 
the species B. amyloliquefaciens, B.

[[Page 13766]]

amyloliquefaciens subsp. amyloliquefaciens and B. amyloliquefaciens 
subsp. plantarum. The former subspecies includes the type strain and 
likely most, if not all, of the industrial strains of B. 
amyloliquefaciens used for enzyme production. The latter subspecies 
consists of plant-associated strains used as biocontrol agents due to 
the production of several antifungal lipopeptide and antibacterial 
polyketide toxins. This exemption is restricted to the subspecies B. 
amyloliquefaciens subsp. amyloliquefaciens which contains the 
industrial strains used for enzyme production. Adequate genotypic and 
phenotypic information is available to accurately identify B. 
amyloliquefaciens subsp. amyloliquefaciens.
    2. Information to evaluate the relationship of the microorganism to 
any other closely related microorganisms which have a potential for 
adverse effects on health or the environment.
    There are several species in the genus Bacillus that are known 
pathogens. These include Bacillus anthracis, which is pathogenic to 
humans and other animals, and Bacillus cereus, which is a common cause 
of food poisoning. Bacillus thuringiensis, Bacillus larvae, Bacillus 
lentimorbus, Bacillus popilliae, and some strains of Bacillus 
sphaericus are pathogenic or toxigenic to certain insects. The new 
subspecies Bacillus amyloliquefaciens subsp. plantarum has been shown 
to exhibit toxicity mainly to plant pathogenic fungi but can also be 
cytotoxic to mammalian cells. It is possible, using polyphasic 
approaches, to differentiate between B. amyloliquefaciens subsp. 
amyloliquefaciens and these other species and subspecies that have the 
potential to adversely affect humans or other organisms. B. 
amyloliquefaciens can be distinguished from the very similar Bacillus 
subtilis by a few phenotypic traits and DNA dissimilarity.
    3. A history of safe commercial use for the microorganism.
    B. amyloliquefaciens subsp. amyloliquefaciens has been used to 
produce commercial enzymes for more than 50 years. It produces 
carbohydrases, proteases, nucleases, xylanases, and phosphatases that 
have applications in the food, brewing, distilling, and textile 
industries.
    For commercial enzyme production, B. amyloliquefaciens subsp. 
amyloliquefaciens is grown in a closed submerged fermentation. In 
submerged fermentation, growth of the microorganism occurs beneath the 
surface of the liquid growth medium. The fermentation broth is a 
defined mixture of carbon and nitrogen sources, minerals, salts, and 
other nutrients that is maintained at optimal pH and temperature. These 
conditions support the active growth and productivity of the organisms. 
Submerged fermentation systems reduce the potential for exposure of 
workers to the production organism and fermentation broth aerosols, 
reduce the potential for contamination of the culture, and make the 
collection of extracellular enzyme simpler and less costly. The 
fermentation process is terminated before the B. amyloliquefaciens 
subsp. amyloliquefaciens organisms enter the stationary growth phase, 
and the production organisms are separated from the fermentation broth 
and inactivated. The enzyme preparation may also be subjected to other 
purification processes.
    B. amyloliquefaciens subsp. amyloliquefaciens has a long history of 
safe use for enzyme production in food and industrial applications with 
no incidences associated with human pathogenicity. In response to a 
petition from the ETA, FDA affirmed that carbohydrase enzyme 
preparations and protease enzyme preparations derived from either 
Bacillus subtilis or Bacillus amyloliquefaciens are GRAS for use as 
direct food ingredients. The European Food Safety Authority (EFSA) has 
put B. amyloliquefaciens on their list of bacteria that have a 
``qualified presumption of safety'' because of a long history of 
apparent safe use in food and feed production. However, it was put on 
the list with a qualifier that only strains of B. amyloliquefaciens 
that do not have toxigenic potential be used.
    One strain of B. amyloliquefaciens has been used as a biopesticide. 
A naturally occurring strain of B. amyloliquefaciens subsp. plantarum 
was registered in 2000 as a biopesticide active ingredient under the 
Federal Insecticide, Fungicide, and Rodenticide Act. It can only be 
used on certain ornamental, non-food plants in greenhouses and other 
closed structures.
    4. Commercial uses indicating that the microorganism products might 
be subject to TSCA.
    It is expected that intergeneric strains of B. amyloliquefaciens 
subsp. amyloliquefaciens would be used to produce enzymes and to 
manufacture other industrial chemicals subject to TSCA. Many enzymes 
produced by B. amyloliquefaciens, particularly [alpha]-amylase, are 
used in laundry detergents and in textile processing. B. 
amyloliquefaciens also makes a surfactant known as surfactin which 
functions as an antibiotic.
    5. Studies which indicate the potential for the microorganism to 
cause adverse effects to health or the environment.
    a. Human health hazards-- i. Pathogenicity. B. amyloliquefaciens is 
not pathogenic to humans. There are no reports in the literature 
associating B. amyloliquefaciens with infection or disease in humans. 
B. amyloliquefaciens has been categorized as a Biosafety Level 1(BSL1) 
microorganism by the Centers for Disease Control and Prevention (CDC). 
BSL1 microorganisms are well-characterized agents not known to 
consistently cause disease in immunocompetent adult humans, and which 
present minimal potential hazard to laboratory personnel and the 
environment. Animal toxicity studies were performed with B. 
amyloliquefaciens strain FZB24 to support its registration as a 
biopesticide. Tests for acute oral toxicity/pathogenicity, acute 
pulmonary toxicity/pathogenicity, and acute injection toxicity/
pathogenicity showed little to no adverse effects, which indicated low 
mammalian toxicity and a lack of pathogenicity/infectivity.
    ii. Toxins and other secondary metabolites. Although another 
species in the genus Bacillus, B. cereus, has the potential to produce 
food poisoning toxins which cause both emetic and diarrheal syndromes, 
and a variety of local and systemic infections, the risk of food-borne 
disease caused by bacilli other than B. cereus is generally considered 
to be negligible because usually only B. cereus has the genes that 
encode food poisoning toxins. Industrial strains of Bacillus species 
belonging to the Bacillus subtilis group, which includes B. 
amyloliquefaciens, do not express B. cereus toxins. In addition, there 
are no reported cases of food poisoning associated with B. 
amyloliquefaciens.
    Some strains of B. amyloliquefaciens have been shown to produce 
bioactive cyclic lipopeptide metabolites such as iturin, surfactin, 
fengycin, and bacillomycin D. These are cyclical lipoprotein 
biosurfactants produced by non-ribosomal peptide synthesis. They have a 
low mammalian toxicity as demonstrated by a lethal dose of 50% 
(LD50) of >2,500 milligram/kilogram (mg/kg) in an acute 
toxicity test of surfactin C, and a no observed adverse effect level of 
500 mg/kg-day in a repeat dose oral gavage study. Some strains of B. 
amyloliquefaciens may also produce the polyketide toxins macrolactin, 
bacillanene, and difficidin. B. amyloliquefaciens also produces the 
protein toxin barnase and the antifungal protein baciamin.

[[Page 13767]]

    There are reports of the isolation of B. amyloliquefaciens from 
water-damaged buildings in which occupants were suffering ill health 
symptoms. Extracts from biomass of isolated strains of Bacillus 
exhibiting antifungal properties were assessed for toxicity endpoints. 
All of the isolated B. cereus and B. amyloliquefaciens strains studied 
showed cytotoxicity as evidenced by inhibition of boar spermatozoa 
motility; however, the B. amyloliquefaciens strains affected boar 
spermatozoa differently from the indoor B. cereus isolates and the 
reference food-poisoning strain.
    The isolation of cytotoxic strains of B. amyloliquefaciens from 
water-damaged buildings is of little concern in relation to the 
exemption of B. amyloliquefaciens subsp. amyloliquefaciens. It is 
important to note that the Bacillus amyloliquefaciens strains studied 
in water-damaged buildings were specifically selected for further study 
because the isolates exhibited antifungal activity. Some of the 
secondary metabolites produced by these strains of B. amyloliquefaciens 
also exhibited cytotoxicity to mammalian cells (i.e., boar 
spermatozoa). However, industrial strains of B. amyloliquefaciens that 
are classified as B. amyloliquefaciens subsp. amyloliquefaciens have 
been shown not to produce most, if not all, of the antifungal and 
antibacterial lipopeptides and polyketides produced by the biocontrol-
type strains. The genome of the type strain of B. amyloliquefaciens DSM 
7\T\ (now B. amyloliquefaciens subsp. amyloliquefaciens) is very 
similar to the genome of the biocontrol strain FZB42 (B. 
amyloliquefaciens subsp. plantarum). However, the latter subspecies had 
genomic islands carrying prophage sequences, transposases, integrases, 
and recombinases that the DSM 7\T\ type strain did not have. The DSM 
7\T\ type strain was shown to have a diminished capacity to non-
ribosomally synthesize secondary metabolites with antifungal and 
antibacterial activities. The DSM 7\T\ type strain could not produce 
the polyketides difficidin or macrolantin, and could not produce 
lipopeptide such as iturin, macrolantin, and other compounds except for 
the compound surfactin.
    Although there are isolated reports of toxin production in several 
antifungal, environmental isolates of B. amyloliquefaciens, the larger 
body of studies available on the safety and toxicity of B. 
amyloliquefaciens strains used industrially for enzyme production (Ref. 
6) indicate that these strains are safe and non-toxic. For example, the 
industrial strains of B. amyloliquefaciens, Bacillus subtilis, and 
Bacillus licheniformis used for large-scale enzyme production did not 
exhibit any cytotoxicity in Chinese hamster ovary tests. In Europe, the 
toxicity of two strains of B. amyloliquefaciens used in the production 
of [alpha]-amylase and bacillolysin was assessed by EFSA's Scientific 
Panel on Additives and Products or Substances used in Animal Feed. The 
panel concluded that the B. amyloliquefaciens production strains 
DSM9553 and DSM9554, when used as a source of extracellular enzyme, do 
not present a toxigenic risk. Given its widespread distribution in the 
environment, its long history of safe use in industrial fermentation, 
the absence of reports on pathogenicity to humans, and the limited 
reports of cytotoxicity, EPA concludes that the use of B. 
amyloliquefaciens in fermentation facilities for production of enzymes 
or specialty chemicals does not present a human health concern.
    b. Environmental hazards-- i. Hazards to animals. There are no 
reports suggesting that B. amyloliquefaciens is pathogenic to 
domesticated animals or wildlife. The cytotoxicity of antifungal 
secondary metabolites to mammalian cells by biocontrol strains of B. 
amyloliquefaciens subsp. plantarum is discussed in this Unit.
    ii. Hazards to plants. B. amyloliquefaciens is not pathogenic to 
plants. The plant-associated strains of B. amyloliquefaciens are 
beneficial to plants because they inhibit the growth of fungal plant 
pathogens. Antifungal and antibacterial secondary metabolites produced 
by strains of B. amyloliquefaciens such as iturins, surfactins, 
fengycin, bacillomycins, and azalomycin have been shown to inhibit the 
growth of Rhizoctonia solani, Xanthomonas campestris pv. campestris, 
Alternaria brassicae, Botyris cinerea, Leptosphaeria maculans, 
Verticillium longisporum, Pythium ultimatum, Aspergillus spp., Fusarium 
spp., Bipolaris sorokiniana, and Fusarium oxysporum.
    In addition to producing antifungal and antibacterial compounds, B. 
amyloliquefaciens is known as a plant growth-promoting rhizobacterium, 
and some of the biological control strains of B. amyloliquefaciens were 
shown to produce the phytohormone indole-3-acetic acid.
    6. Studies which indicate the survival characteristics of the 
microorganism in the environment.
    Several studies assessing the survival of B. amyloliquefaciens are 
available in the public literature and are described in EPA's Risk 
Assessment of B. amyloliquefaciens (Ref. 6). Given that the natural 
habitat for B. amyloliquefaciens is typically in soil, on plant roots, 
or as an endophyte within the roots or stems of plants, the bacterium 
is likely to survive for a least some period of time if inadvertently 
released to the environment. However, like other bacilli, survival in 
soil may occur predominately as the resistant endospore state, whereas 
in the rhizosphere, it may exist as active vegetative cells.

VI. Physical Containment and Control Technologies for T. reesei QM6a 
and B. amyloliquefaciens subsp. amyloliquefaciens

A. Release and Exposure Assessment in Support of the TSCA Section 
5(h)(4) Exemption for T. reesei QM6a

    The estimated releases of the microorganism from an enzyme 
manufacturing facility and exposures of workers, the general 
population, and the environment to the microorganisms are based on a 
generic scenario developed by EPA in 1997 for large-scale closed system 
enzyme fermentation. The generic scenario assumes the facility operates 
350 days/year, produces 100 batches/year, the maximal cell 
concentration in the fermentation broth is 1 x 10\7\ colony-forming 
units (cfu)/ml, and the volume of the fermentation broth is 70,000 L. 
The process consists of the main steps of laboratory propagation, 
fermentation, inactivation, and recovery where filtration operations 
separate out the microbial biomass from the concentrated desired 
product. The operations, sources of exposure and release are described 
in more detail in EPA's Release and Exposure Assessment (Ref. 13).
    Exposures of workers to the microorganisms in during processing 
operations using submerged standard industrial fermentation do not pose 
concerns. The release of microbial cells in aerosols or in liquid and 
solid waste streams in submerged standard industrial fermentation 
operations with the containment and inactivation conditions of the Tier 
I exemption, are considered low. Thus, potential exposures to the 
general human population to the microorganism through inhalation or 
drinking water ingestion and to the environment are also low.

[[Page 13768]]

B. Release and Exposure Assessment in Support of the TSCA Section 
5(h)(4) Exemption for B. amyloliquefaciens subsp. amyloliquefaciens

    The estimated releases of the microorganism from an enzyme 
manufacturing facility and exposures of the microorganisms to workers, 
the general population, and the environment are based on a generic 
scenario developed by EPA in 1997 for large-scale closed system enzyme 
fermentation. The generic scenario assumes the facility operates 350 
days/year, produces 100 batches/year, the maximal cell concentration in 
the fermentation broth is 1 x 10\11\ cfu/ml and the volume of the 
fermentation broth is 70,000 L. The process consists of the main steps 
of laboratory propagation, fermentation and then recovery where 
filtration operations separate out the biomass from the concentrated 
desired product. The operations, sources of exposure and release are 
described in more detail in EPA's Release and Exposure Assessment (Ref. 
14).
    Exposures of workers to the microorganisms during processing 
operations using submerged standard industrial fermentation do not pose 
concerns. The release of microbial cells in aerosols or in liquid and 
solid waste streams in submerged standard industrial fermentaion 
operations with the containment and inactivation conditions of the Tier 
I exemption are considered low. Thus, potential exposures to the 
general human population to the microorganism through inhalation or 
drinking water ingestion and to the environment are also low.

VII. Risk Assessment Overview for T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens

    EPA's Risk Assessment documents (Refs. 5 and 6) provide more 
detailed information, and supporting references, for EPA's evaluation 
of the available information and the potential risks to health and the 
environment. EPA has determined that because of the low hazard 
potential and safe history of use of T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens, the TSCA section 5(h)(4) 
exemption will not present an unreasonable risk of injury to health or 
the environment, including an unreasonable risk of injury to a 
potentially exposed or susceptible subpopulation under the conditions 
of use, provided that the other conditions of the exemptions at 40 CFR 
part 725, subpart G, relating to the introduced genetic material, and 
the physical containment of the new microorganisms, have been met.

A. Risk Assessment for T. reesei QM6a

    There is only one potential concern for human health and 
environmental hazards associated with T. reesei QM6a, and that is for 
paracelsin production. Paracelsin production is not expected to occur 
in submerged standard industrial fermentation operations conducted 
solely for growth of the microorganism to produce enzymes. There is no 
concern for potential pathogenicity of T. reesei QM6a to humans, 
plants, domesticated animals, or wildlife. The body of evidence of 
pathogenicity testing on various industrial strains indicates that T. 
reesei is not pathogenic to humans. Toxicity testing on a number of 
enzymes produced by T. reesei indicates that the fungus does not 
produce toxins when used in the submerged standard industrial 
fermentation operations used for enzyme production.
    T. reesei has a long history of safe use and is expected to present 
low hazard to workers, the general public, and the environment. 
Although direct monitoring data are unavailable, estimates of potential 
exposures made by EPA in its assessment of potential risks (Ref. 5) do 
not indicate high levels of exposure of T. reesei to either workers or 
the public from submerged standard industrial fermentation operations 
used for enzyme production. Standard industrial hygiene management 
practices currently used in the fermentation industry reduce the 
potential for adverse health effects in the workplace. The use of 
engineering controls (closed fermentation systems), appropriate work 
practices, personal protective equipment, and personal hygiene reduce 
the potential for worker exposure. Thus, current practices reduce the 
potential for the dermal and respiratory exposures estimated by EPA.
    Based on worst-case exposure scenarios and toxicity of the 
microorganism, EPA has made the determination that the potential risk 
to workers, the general public, and to the environment resulting from 
the use of T. reesei QM6a in submerged standard industrial fermentation 
operations used for enzyme production is low, provided the additional 
criteria of the tiered exemptions for the introduced genetic material 
and the physical containment conditions are met (Ref. 5).

B. Risk Assessment for B. amyloliquefaciens subsp. amyloliquefaciens

    Industrial strains of B. amyloliquefaciens subsp. amyloliquefaciens 
are not pathogenic to humans, plants, domesticated animals, or 
wildlife, and do not produce many of the toxic secondary metabolites 
found in biological control strains of B. amyloliquefaciens subsp. 
plantarum. The long history of safe use of enzymes produced by 
industrial strains of B. amyloliquefaciens in food is evidence that the 
bacterium does not produce toxins under standard conditions used for 
enzyme production.
    Current practices in the fermentation industry reduce the potential 
for adverse health effects in the workplace. The use of engineering 
controls (closed fermentation systems), appropriate work practices, 
personal protective equipment, and personal hygiene reduce the 
potential for worker exposure and reduce the potential for the dermal 
and respiratory exposures.
    Industrial strains of B. amyloliquefaciens have a long history of 
safe use and are expected to present low hazard to workers, the general 
public, and the environment. Although direct monitoring data are 
unavailable, exposure estimates do not suggest high levels of exposure 
of B. amyloliquefaciens subsp. amyloliquefaciens to either workers or 
the public resulting from the industrial fermentation procedures that 
are standard throughout the industry.
    Based on worst-case exposure scenarios and toxicity of the 
microorganism, EPA has made the determination that the potential risk 
to workers, the general public, and the environment associated with the 
use of industrial strains of B. amyloliquefaciens subsp. 
amyloliquefaciens in submerged standard industrial fermentation is low 
provided the additional criteria of the tiered exemptions for the 
introduced genetic material and the physical containment conditions are 
met (Ref. 6).

VIII. Rationale for Adding T. reesei QM6a and B. amyloliquefaciens 
subsp. amyloliquefaciens to the List of Recipient Microorganisms at 40 
CFR 725.420

 A. Statutory Background

    On June 22, 2016, the ``Frank R. Lautenberg Chemical Safety for the 
21st Century Act,'' amended TSCA (15 U.S.C. 2601 et seq.) (Ref. 15). 
Pursuant to TSCA section 5(h)(4), EPA is authorized, upon request and 
by rule, to exempt the manufacturer of any new

[[Page 13769]]

chemical substance from all or part of the requirements of TSCA section 
5 if EPA determines that the manufacture, processing, distribution in 
commerce, use, or disposal of the chemical substance, or any 
combination of such activities, will not present an unreasonable risk 
of injury to human health or the environment, including an unreasonable 
risk to a potentially exposed or susceptible subpopulation identified 
by the Administrator under the conditions of use. The amended language 
of the statute with regard to section 5(h)(4) did not alter EPA's 
approach to balancing the considerations of the costs and benefits of 
issuing an exemption rule.

B. EPA's Approach for Assessing ``Unreasonable Risk'' for T. reesei 
QM6a and B. amyloliquefaciens subsp. amyloliquefaciens

    In determining whether T. reesei QM6a and B. amyloliquefaciens 
subsp. amyloliquefaciens will not present an unreasonable risk of 
injury to human health or the environment, the Agency considered more 
than just the inherent risks presented by the two microorganisms. The 
Agency also considered the full range of societal benefits associated 
with the exemption; for example, as discussed in more detail below, EPA 
considered not only the cost savings to the users of the microorganism, 
but also the societal benefits that flow from promotion of the use of 
low-risk recipient microorganisms, while allowing the Agency to direct 
its resources toward reviewing higher risk microorganisms.
    It is important that EPA is revising one aspect of the existing 
tiered exemptions at 40 CFR 725.420 by expanding the exemption to apply 
to two specific microorganisms. The narrow scope of this action 
affected the scope of EPA's cost-benefit analysis in which EPA compared 
the risks and benefits of the two microorganisms being considered for 
an exemption with the risks that would have resulted if those same two 
microorganisms remained subject to full MCAN submission requirements 
and 90-day EPA review. EPA did not compare the risks and benefits that 
would result from use of these two microorganisms in the absence of any 
regulation.
    It is also significant that the standard applicable to this rule is 
that the microorganisms ``will not present unreasonable risk,'' rather 
than ``no risk.'' It is not possible to eliminate all risks associated 
with the manufacture, processing, distribution in commerce, use, and 
disposal of any new microorganism.

C. Application of No Unreasonable Risk Factors for T. reesei QM6a and 
B. amyloliquefaciens subsp. amyloliquefaciens

    The following is an explanation of the factors and their analyses 
relevant to the no unreasonable risk finding.
    1. Risks associated with these two microorganisms. EPA's evaluation 
of the available information concerning T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens against these criteria is 
presented in detail in Unit V., and is summarized again here for the 
readers' convenience.
    The Agency developed specific criteria in 40 CFR 725.67 that the 
Agency uses in determining the extent of a potential recipient 
microorganism's risks, and its eligibility for listing at 40 CFR 
725.420. These criteria were explained in detail in the proposed 
``biotech'' rule (Ref. 16), the final ``biotech'' rule (Ref. 17), and 
are discussed in Unit V. EPA's conclusions for these two microorganisms 
are based on the available data and EPA's experience under 40 CFR part 
725. T. reesei QM6a is not pathogenic to humans, plants, domesticated 
animals, or wildlife and the fungus does not produce toxins under 
submerged standard industrial fermentation operations used for enzyme 
production. T. reesei QM6a has a long history of safe use and is 
generally expected to present low risk to workers, the general public, 
and the environment resulting from submerged standard industrial 
fermentation operations used for enzyme production that are standard 
throughout the industry.
    Under non-standard conditions of fermentation, such as with the 
deliberate fermentation of cellulosic biomass for saccharification of 
plant material or extended fermentation, paracelsin may be produced. 
The risks associated with the production of paracelsin may be 
significant due to its toxicity to mammalian cells, aquatic species, 
Gram-positive bacteria, and various fungi. However, the potential risk 
associated with paracelsin production is expected to be significantly 
reduced by this rule, which limits the exemption to fermentation 
operations using submerged standard industrial fermentation operations 
used for enzyme production.
    Industrial strains of B. amyloliquefaciens subspecies 
amyloliquefaciens are not pathogenic to humans, plants, domesticated 
animals, or wildlife, and do not produce toxins under standard 
conditions used for enzyme production. Industrial strains of B. 
amyloliquefaciens subsp. amyloliquefaciens used for the production of 
enzymes have a long history of safe use and are expected to present low 
hazards to human health and the environment.
    Only strains of B. amyloliquefaciens that fall into the subspecies 
B. amyloliquefaciens amyloliquefaciens were considered as the eligible 
recipient microorganism at 40 CFR 725.420. In this rule, EPA is 
excluding other strains/subspecies of these two species for which:
     The Agency has insufficient data and review experience to 
find that they will not present an unreasonable risk of injury or
     The Agency has found that, under certain conditions, based 
on data on the species in question, a strain or subspecies may present 
an unreasonable risk, thereby requiring a closer examination of the 
conditions of manufacturing, processing, distribution in commerce, use, 
and disposal during a 90-day MCAN review. Consequently, additional 
information would be necessary to make an appropriate determination 
about the organisms' potential risks.
    The Agency believes that the requirement for submission of a MCAN 
followed by a 90-day review period for new intergeneric microorganisms 
that use T. reesei QM6a and B. amyloliquefaciens subsp. 
amyloliquefaciens as recipient microorganisms is not necessary to 
address the risks associated with these microorganisms and would not 
result in any additional protection than would be achieved by this 
rule. This conclusion is based, in part, on EPA's findings regarding 
the intrinsically low level of hazard that these two organisms pose to 
human health and the environment. The requirements of the Tier I and 
Tier II exemptions and the restrictions in this rule on fermentation 
conditions place sufficient constraints to significantly limit the 
potential risks of injury to human health or the environment, including 
potential risks to potentially exposed or susceptible subpopulations 
under the conditions of use. In making this determination, EPA 
identified workers as a potentially exposed or susceptible 
subpopulation to the substance under the conditions of use and 
concluded that, with the limitations described above, the substances 
will not present an unreasonable risk of injury to health or the 
environment.
    The Agency concludes that the criteria set forth in this rule are 
sufficient to mitigate the identified risks associated with these 
microorganisms. Because of the low hazard potential and safe history of 
use of T. reesei QM6a and

[[Page 13770]]

B. amyloliquefaciens subsp. amyloliquefaciens, EPA concludes that the 
TSCA section 5(h)(4) exemption will not present an unreasonable risk of 
injury to health or the environment, including an unreasonable risk of 
injury to a potentially exposed or susceptible subpopulation under the 
conditions of use, provided that the other conditions of the exemptions 
at 40 CFR part 725, subpart G, relating to the introduced genetic 
material, and the physical containment of the new microorganisms, have 
been met.
    2. Costs. As discussed in Unit X., this rule is anticipated to 
reduce costs to currently regulated entities in the long run. Expanding 
the list of recipient microorganisms eligible for exemption does not 
otherwise impose any additional cost or other burden on currently 
regulated entities, or existing fermentation processes.
    Limiting the use of this exemption to the identified fermentation 
conditions is also estimated to impose no burden on affected entities. 
The restriction merely codifies existing industrial fermentation 
procedures for manufacturing operations that currently seek to use 
tiered exemptions. Consequently, EPA expects that most, if not all, 
manufacturers using these microbes would already have the measures in 
place to qualify for the exemption. Equally important, this limitation 
would add no burden to any existing fermentation processes. Currently, 
fermentation operations with either of these microbes are not eligible 
for the tiered exemption, and thus a MCAN must be submitted. Any 
company that chooses to use a different fermentation process could 
continue to operate under the status quo and simply submit a MCAN. This 
rule simply offers an additional, less costly option, to facilities 
that choose to use the fermentation operations discussed in this rule.
    3. Benefits. The following discussion describes the benefits of 
expanding the list of recipient microorganisms eligible for exemption 
in a qualitative manner; for a more quantitative approach, see the 
economic analysis prepared for this rule (Ref. 18). A summary of that 
economic analysis is also provided in Unit IX.
    The benefits analyzed encompass more than the direct benefits 
associated with submitting a Tier I or Tier II exemption for a new 
intergeneric microorganism rather than a MCAN. EPA's benefit analysis 
included a consideration of the broader benefits to society. EPA's 
unreasonable risk determination is based on broader benefits to society 
as well as those benefits attributable to a reduction in the burden 
associated with submission of Tier I and Tier II exemptions rather than 
MCANs.
    EPA has concluded that manufacturers of new intergeneric 
microorganisms based on these low-risk microorganisms currently bear an 
unnecessary regulatory burden. By adding T. reesei QM6a and B. 
amyloliquefaciens subsp. amyloliquefaciens to the list of eligible 
recipient microorganisms in 40 CFR 725.420, the Agency removes 
unnecessary regulatory impediments to the design, manufacture, and 
commercialization of these low risk new intergeneric microorganisms, 
and of the chemical substances that can be produced by these safer 
microorganisms. This action will also reduce the costs associated with 
industry's reporting burden, including the costs associated with the 
preparation of the submission, and with the delay in the commercial 
market introduction of the new intergeneric microorganism. Some of the 
cost-savings benefits may accrue to small businesses, either as 
developers of the exempt microorganisms, as producers of fermentation 
chemicals using the live microorganisms, or as customers for enzymes or 
other products made using the microorganisms.
    There will also be a reduction in the Agency review resources 
currently allocated to reviews of MCANs for these two microorganisms. 
These Agency resources will be shifted to the review of new 
intergeneric microorganisms or chemical substances of greater concern.
    The addition of the two microorganisms to the list of 
microorganisms eligible for exemption is expected to encourage 
innovation in the industry. It is reasonable to assume that a new 
intergeneric microorganism would either possess a new function or serve 
an existing function more efficiently or at a lower cost. The reduction 
in delay for that new intergeneric microorganism to be introduced into 
commerce is expected to be a benefit to both manufacturers and the 
general public who will have access to the substance more quickly. The 
expected benefits to innovation have not been quantified but include: 
Reduced time to develop and commercialize organisms; decreased cost of 
some downstream industrial products, such as fuel ethanol; improved 
consumer appeal of some products, such as certain textiles; and reduced 
costs of some consumer products, such as detergent and leather goods.
    4. Risk/benefit balance. Determining the presence or absence of an 
unreasonable risk for purposes of issuing an exemption pursuant to TSCA 
section 5(h)(4) requires balancing of the benefits and risks posed by a 
regulatory action. EPA has determined that the risks are generally low 
based on the inherent properties and intended uses of T. reesei QM6a 
and B. amyloliquefaciens subsp. amyloliquefaciens and will be 
adequately managed by the restrictions in the rule, combined with the 
existing requirements of the Tier I and Tier II exemptions.
    EPA anticipates that expanding the list of microorganisms eligible 
for exemption will impose no costs and will reduce costs to currently 
regulated entities that use those recipients. The limitation to certain 
fermentation conditions is not a cost that will be imposed by this rule 
but rather a limitation on the amount of regulatory relief it will 
provide. The limitations on fermentation conditions reflect industrial 
fermentation procedures that are currently common practices for the 
affected industry.
    EPA has also concluded that the benefits of the addition of T. 
reesei QM6a and B. amyloliquefaciens subsp. amyloliquefaciens as 
recipient microorganisms to the list of recipient microorganisms at 40 
CFR 725.420 are quite significant. This addition reduces the overall 
regulatory burden for affected entities by reducing the reporting 
requirements and by eliminating the delay of these products into 
commerce. The rule benefits both regulated entities and the general 
public by promoting the expedited manufacture and use of the chemical 
substances produced using these low-risk organisms and manufacturing 
processes. There is also the added benefit of concentrating limited EPA 
resources on regulation of chemical substances which have a greater 
potential to present significant risks, rather than on these two 
microorganisms. While this is difficult to quantify, it is considered 
substantial.
    In sum, the criteria set forth in this exemption are sufficient to 
mitigate the low level of potential risks presented by these organisms, 
particularly when compared to the benefits, in toto, of this exemption, 
to levels that are consistent with the statutory standard for an 
exemption. Consequently, EPA has determined that adding T. reesei QM6a 
and B. amyloliquefaciens subsp. amyloliquefaciens as recipient 
microorganisms to the list of recipient microorganisms at 40 CFR 
725.420 is appropriate. The two microorganisms

[[Page 13771]]

will not present an unreasonable risk of injury to human health or the 
environment when manufactured under the conditions of this exemption.

IX. Economic Impacts

    EPA's economic analysis (Ref. 18) evaluates the potential for 
significant economic impacts as a result of the addition of two 
microorganisms (T. reesei QM6a and B. amyloliquefaciens subsp. 
amyloliquefaciens) to 40 CFR 725.420, which lists recipient 
microorganisms eligible for Tier I and Tier II exemptions. Over the 
course of the first 10 years after the effective date of the final 
rule, EPA estimates that the addition of the two microorganisms to the 
list will generate a total cost savings to society of approximately 
$4.5 million. Industry is estimated save approximately $2.7 million and 
the Agency approximately $1.8 million. The equivalent, annualized cost 
savings to industry are expected to be $260,000 per year and $252,000 
per year at a 3% and 7% discount rate, respectively. EPA estimates that 
there will be a net decrease in burden to industry of 27,864 hours over 
this 10-year period.

X. Scientific Standards, Evidence, and Available Information

    EPA has used scientific information, technical procedures, 
measures, methods, protocols, methodologies, and models consistent with 
the best available science, as applicable. These sources supply 
information relevant to a determination that the microorganisms subject 
to this rule will not present an unreasonable risk of injury to health 
or the environment, including an unreasonable risk to a potentially 
exposed or susceptible subpopulation identified by the Administrator 
under the conditions of use. The clarity and completeness of the data, 
assumptions, methods, quality assurance, and analyses employed are 
documented, as applicable and to the extent necessary for purposes of 
this rule, in Units V. through VIII. and in the references. The extent 
to which the various information, procedures, measures, methods, 
protocols, methodologies or models used in EPA's decision have been 
subject to independent verification or peer review is adequate to 
justify their use, collectively, in the record for this rule.

XI. References

    The following is a listing of the documents that are specifically 
referenced in this document. The docket includes these documents and 
other information considered by EPA, including documents that are 
referenced within the documents that are in the docket, even if the 
referenced document is not physically located in the docket. For 
assistance in locating these other documents, please consult the 
technical person listed under FOR FURTHER INFORMATION CONTACT.
    1. Genencor International, Inc. Letter of Application to list 
Trichoderma reesei as exempt under subpart G of 40 CFR part 725--
Reporting Requirements and Review Processes for Microorganisms. 
March 17, 2005.
    2. Novo Nordisk BioChem North America, Inc. Letter of 
Application to list Bacillus amyloliquefaciens as exempt under 
subpart G of 40 CFR part 725--Reporting Requirements and Review 
Processes for Microorganisms. November 7, 1997.
    3. EPA, OPPT. Email confirming Novo Nordisk BioChem North 
America, Inc.'s letter of application to list Bacillus 
amyloliquefaciens as exempt under subpart G of 40 CFR part 725--
Reporting Requirements and Review Processes for Microorganisms. 
August 3, 2009.
    4. US EPA. Microorganisms; General Exemptions from Reporting 
Requirements; Revisions to Recipient Organisms Eligible for Tier I 
and Tier II Exemptions; Proposed Rule. RIN 2070-AJ65; FRL-9348-1. 77 
FR 54499, September 5, 2012. (``2012 Proposed Rule'').
    5. EPA, OPPT. Risk Assessment of Trichoderma reesei for 
Consideration of Addition to the List of Eligible Recipient 
Microorganisms for the 5(h)(4) Exemptions from MCAN Reporting 
Requirements. October 2011.
    6. EPA, OPPT. Risk Assessment of Bacillus amyloliquefaciens 
subsp. amyloliquefaciens for Consideration of Addition to the List 
of Eligible Recipient Microorganisms for the 5(h)(4) Exemptions from 
MCAN Reporting Requirements. July 2015.
    7. Anonymous Public Comment, Document ID: EPA-HQ-OPPT-2011-0740-
0015; October 23, 2012.
    8. Richard Fitti, West Chester University of PA Comment, 
Document ID: EPA-HQ-OPPT-2011-0740-0017; November 5, 2012.
    9. Anthony T. Pavel, General Counsel & Secretary, Enzyme 
Technical Association (ETA) and Rina Singh, Director of Policy, 
Science & Renewable Chemicals, Industrial and Environmental Section, 
Biotechnology Industry Organization (BIO) Comment, Document ID EPA-
HQ-OPPT-2011-0740-0016; November 2012.
    10. ETA. Supplemental information on Trichoderma reesei. January 
29, 2010.
    11. ETA. Supplemental information on Trichoderma reesei. June 
16, 2011.
    12. Nevalainen, H., P. Suominen, K. Tasimisto. 1994. On the 
safety of Trichoderma reesei. J. Biotechol. 37:193-200.
    13. EPA, OPPT. Release and Exposure Assessment in Support of the 
TSCA Section 5(h)(4) Exemption for Trichoderma reesei. June 2011.
    14. EPA, OPPT. Release and Exposure Assessment in Support of the 
TSCA Section 5(h)(4) Exemption for Bacillus amyloliquefaciens. June 
2011.
    15. Legislative History of the Toxic Substances Control Act, pp. 
409-423. House Report 1341, 94th Congress, 2nd Session. 1976.
    16. EPA. Microbial Products of Biotechnology; Proposed 
Regulation under the Toxic Substances Control Act. Federal Register 
(59 FR 45526; September 1, 1994) (FRL-4774-4).
    17. EPA. Microbial Products of Biotechnology; Final Regulation 
under the Toxic Substances Control Act. Federal Register (62 FR 
17910; April 11, 1997) (FRL-5577-2).
    18. EPA, OPPT. Economic Analysis for the Final Biotechnology 
Exemptions Rule for Trichoderma reesei and Bacillus 
amyloliquefaciens. October 2019.

XII. Statutory and Executive Order Reviews

    Additional information about these statutes and Executive Orders 
can be found at http://www2.epa.gov/laws-regulations/laws-and-executive-orders.

A. Executive Order 12866: Regulatory Planning and Review and Executive 
Order 13563: Improving Regulations and Regulatory Review

    This action is not a significant regulatory action and was 
therefore not submitted to the Office of Management and Budget (OMB) 
for review under Executive Orders 12866 (58 FR 51735, October 4, 1993) 
and 13563 (76 FR 3821, January 21, 2011).

B. Executive Order 13771: Reducing Regulations and Controlling 
Regulatory Costs

    This is considered a deregulatory action under Executive Order 
13771 (82 FR 9339, February 3, 2017) because this rule is expected to 
provide meaningful burden reduction by adding T. reesei and B. 
amyloliquefaciens subspecies amyloliquefaciens to the list of recipient 
microorganisms that may be used to qualify for the Tier I and Tier II 
exemptions from full notification and reporting under TSCA for new 
microorganisms that are being manufactured for introduction into 
commerce. The rule is expected to generate cost savings for 
organizations that, in the absence of the rule, would submit MCANs for 
new intergeneric T. reesei or B. amyloliquefaciens strains. EPA 
estimates that the rule will result in cost savings for both industry 
and the Agency.

C. Paperwork Reduction Act (PRA)

    This action does not impose any new information collection 
requirements or related burden that would require additional review or 
approval by OMB under the PRA, 44 U.S.C. 3501 et seq. The information 
collection activities associated with the submission of Tier 1 and Tier 
2 notices under TSCA have already been approved by OMB

[[Page 13772]]

pursuant to the PRA and are covered by the following existing 
Information Collection Requests (ICRs): OMB control numbers 2070-0012 
(EPA ICR No. 0574.15) and 2070-0038 (EPA ICR No. 1188.11). In granting 
these exemptions, this rule does not impose any new information 
collection requirements and is expected to reduce the amount of 
required reporting by allowing firms to submit less information for 
qualifying microorganisms. Over the ten-year period, industry is 
expected to subtract a total of 27,864 hours at an average of 2,786 
hours per year.

D. Regulatory Flexibility Act (RFA)

    Pursuant to section 605(b) of the RFA, 5 U.S.C. 601 et seq., I 
certify that this final rule will not have a significant economic 
impact on a substantial number of small entities. In making this 
determination, EPA believes that the impact of concern is any adverse 
economic impact on small entities, and that EPA may certify that a rule 
will not have a significant economic impact on a substantial number of 
small entities if the rule relieves regulatory burden, has no net 
burden or otherwise has a positive economic effect on the small 
entities subject to the rule. This action establishes exemptions from 
existing requirements that apply regardless of the size of the entity. 
The factual basis for this certification is presented in the small 
entity impact analysis that was prepared as part of the Economic 
Analysis for this rule (Ref. 18) and is briefly summarized in Unit 
VIII.

E. Unfunded Mandates Reform Act (UMRA)

    This action does not contain an unfunded mandate as described in 
UMRA, 2 U.S.C. 1531-1538, and does not significantly or uniquely affect 
small governments. This action is not expected to impose enforceable 
duty on any state, local or tribal governments, and the requirements 
imposed on the private sector are not expected to result in annual 
expenditures of $100 million or more for the private sector. As such, 
EPA has determined that the requirements of UMRA sections 202, 203, 
204, or 205 do not apply to this action.

F. Executive Order 13132: Federalism

    This action does not have federalism implications as specified in 
Executive Order 13132 (64 FR 43255, August 10, 1999). It will not have 
substantial direct effects on the states, on the relationship between 
the national government and the states, or on the distribution of power 
and responsibilities among the various levels of government. EPA has no 
information to indicate that any state or local government commercially 
manufactures or processes the microorganisms covered by this action. 
Thus, Executive Order 13132 does not apply to this action.

G. Executive Order 13175: Consultation and Coordination With Indian 
Tribal Governments

    This action does not have tribal implications as specified in 
Executive Order 13175 (65 FR 67249, November 9, 2000). It will not have 
substantial direct effects on tribal governments, on the relationship 
between the Federal government and the Indian tribes, or on the 
distribution of power and responsibilities between the Federal 
government and Indian tribes. EPA has no information to indicate that 
any tribal government commercially manufactures or processes the 
microorganisms covered by this action. Thus, E.O. 13175 does not apply 
to this action.

H. Executive Order 13045: Protection of Children From Environmental 
Health Risks and Safety Risks

    EPA interprets Executive Order 13045 (62 FR 19885, April 23, 1997), 
as applying only to those regulatory actions that concern health or 
safety risks, such that the analysis required under section 5-501 of 
Executive Order 13045 has the potential to influence the regulation. 
This action is not subject to Executive Order 13045 because it does not 
establish an environmental standard intended to mitigate health or 
safety risks.

I. Executive Order 13211: Actions That Significantly Affect Energy 
Supply, Distribution, or Use

    This action is not a ``significant energy action'' as defined in 
Executive Order 13211 (66 FR 28355, May 22, 2001), because it is not a 
significant regulatory action under Executive Order 12866, and is not 
likely to have a significant adverse effect on energy supply, 
distribution, or use.

J. National Technology Transfer and Advancement Act (NTTAA)

    Since this action does not involve any technical standards, NTTAA 
section 12(d), 15 U.S.C. 272 note, does not apply to this action.

K. Executive Order 12898: Federal Actions to Address Environmental 
Justice in Minority Populations and Low-income Populations

    This action does not entail special considerations of environmental 
justice related issues as delineated by Executive Order 12898 (59 FR 
7629, February 16, 1994), because it does not establish an 
environmental health or safety standard.

VII. Congressional Review Act (CRA)

    This action is subject to the CRA, 5 U.S.C. 801 et seq., and EPA 
will submit a rule report to each House of the Congress and to the 
Comptroller General of the United States. This action is not a ``major 
rule'' as defined by 5 U.S.C. 804(2).

List of Subjects in 40 CFR Part 725

    Environmental protection, Administrative practice and procedure, 
Biotechnology, Chemicals, Hazardous substances, Imports, Labeling, 
Microorganisms, Occupational safety and health, Reporting and 
recordkeeping requirements.

    Dated: March 4, 2020.
Alexandra Dapolito Dunn,
Assistant Administrator, Office of Chemical Safety and Pollution 
Prevention.

    Therefore, 40 CFR chapter I is amended as follows:

PART 725--[AMENDED]

0
 1. The authority citation for part 725 continues to read as follows:

    Authority: 15 U.S.C. 2604, 2607, 2613, and 2625.

0
 2. In Sec.  725.3, add in alphabetical order a definition for 
``Submerged standard industrial fermentation'' to read as follows:

Sec.  725.3  Definitions.

* * * * *
    Submerged standard industrial fermentation means a fermentation 
system that meets all of the following conditions:
    (1) Enzyme production is conducted under conditions of submerged 
fermentation (i.e., growth of the microorganism occurs beneath the 
surface of the liquid growth medium).
    (2) Any fermentation of solid plant material or insoluble 
substrates, to which T. reesei fermentation broth is added after the 
submerged standard industrial fermentation operations used for enzyme 
production is completed, may be initiated only after the inactivation 
of the microorganism as delineated in 40 CFR 725.422(d).
* * * * *

0
3. In Sec.  725.420, add paragraphs (k) and (l) to read as follows:

Sec.  725.420  Recipient microorganisms.

* * * * *
    (k) Trichoderma reesei strain QM6a and its derivatives used only in

[[Page 13773]]

submerged standard industrial fermentation operations as defined at 40 
CFR 725.3.
    (l) Bacillus amyloliquefaciens subsp. amyloliquefaciens.

[FR Doc. 2020-04746 Filed 3-9-20; 8:45 am]
 BILLING CODE 6560-50-P