Document ID: EPA-HQ-OPP-2016-0566-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2016-11-30T05:00Z

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EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY NOTICE OF
FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER  

EPA Biopesticides and Pollution Prevention Division contact: [Michael
Glikes, 703-305-6231]

INSTRUCTIONS:  Please utilize this outline in preparing the pesticide
petition.  In cases where the outline element does not apply, please
insert “NA-Remove” and maintain the outline. Please do not change
the margins, font, or format in your pesticide petition. Simply replace
the instructions that appear in green, i.e., “[insert company
name],” with the information specific to your action.

SUBMISSION: E-mail the completed template to: hollis.linda@epa.gov.

TEMPLATE:

Arizona Cotton Research and Protection Council

3721 East Wier Avenue

Phoenix, Arizona  85040-2933

[6E8471]

	EPA has received a pesticide petition (6E8471) from The IR-4 Project,
500 College Road East, Suite 201 W, Princeton, NJ 08540 on behalf of
Arizona Cotton Research and Protection Council, 3721 East Wier Avenue,
Phoenix, Arizona 85040-2933, proposing, pursuant to section 408(d) of
the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to
amend 40 CFR part 180 to establish an exemption from the requirement of
a tolerance for Aspergillus flavus AF36, a non-aflatoxin-producing
strain, in or on almonds and figs.

	Pursuant to section 408(d)(2)(A)(i) of  FFDCA, as amended, The IR-4
Project on behalf of Arizona Cotton Research and Protection Council has
submitted the following summary of information, data, and arguments in
support of their pesticide petition. This summary was prepared by The
IR-4 Project on behalf of Arizona Cotton Research and Protection
Council, 3721 East Wier Avenue, Phoenix, Arizona 85040-2933 and EPA has
not fully evaluated the merits of the pesticide petition. The summary
may have been edited by EPA if the terminology used was unclear, the
summary contained extraneous material, or the summary unintentionally
made the reader conclude that the findings reflected EPA’s position
and not the position of the petitioner.

I. [Interregional Research Project Number 4 (IR-4), Rutgers University,
500 College Road East, Suite 201W, Princeton, NJ 08540 on behalf of the
Arizona Cotton Research and Protection Council, 3721 East Wier Avenue,
Phoenix, Arizona 85040-2933]  Petition Summary

	[6E8471]

A. Product Name and Proposed Use Practices

Aspergillus flavus AF36, a non-aflatoxin-producing strain of A. flavus,
is proposed for application to almonds and figs to reduce the incidence
of aflatoxin producing strains of Aspergillus flavus and thereby reduce
aflatoxin contamination of almonds and figs. 

Aspergillus flavus AF36 is currently registered for use on cotton, corn
and pistachio.  There is also a time-limited exemption from the
requirement of a tolerance established for residues of Aspergillus
flavus AF36, in or on dried figs, resulting from use of the pesticide
pursuant to a FIFRA section 18 emergency exemption.  The purpose of this
amendment is to register Aspergillus flavus AF36 on almonds and figs.  

B. Product Identity/Chemistry

Identity of the pesticide and corresponding residues. The pesticide and
corresponding residues are identified as Aspergillus flavus AF36.

Magnitude of residues at the time of harvest and method used to
determine the residue.  Aspergillus flavus AF36 is a naturally occurring
fungus that was originally isolated from cottonseed produced in the Yuma
Valley of Arizona.  Aspergillus flavus AF36 has been shown to be
naturally and consistently associated with almonds and figs. 
Aspergillus flavus is a widespread fungus.  It is particularly well
adapted to the hot regions of the U.S. where it is widespread in the
environment. The communities of Aspergillus flavus in the desert and in
agricultural fields are naturally composed of both aflatoxin producing
(toxigenic) and aflatoxin non-producing (atoxigenic) strains.  Both
atoxigenic and toxigenic strains have been found on essentially all
plant material and soils in the Southwestern U.S.  The goal of
applications is to increase the percent of the Aspergillus flavus
(atoxigenic) community composed of the strain AF36 and to decrease the
percent of Aspergillus flavus that produces aflatoxins on the crop and
the field.  An exemption from the requirement of a tolerance is proposed
for almonds and figs.]

A statement of why an analytical method of detecting and measuring the
levels of the pesticide residue are not needed. 

An analytical method for residues is not applicable, as an exemption
from the requirement of a tolerance is proposed for almonds and figs.  

C. Mammalian Toxicological Profile

Acute oral toxicity/pathogenicity study

An acute oral toxicity test was performed whereby a single oral dose of
5,000 milligrams/kilogram (mg/kg) per animal of Aspergillus flavus AF36
colonized wheat seed was administered by gavage to five male and five
female Sprague Dawley rats.  The oral LD50 of Aspergillus flavus AF36
was determined to be greater than 5,000 mg/kg rat body weight.  No
clinical signs were observed during the 14 day study and no
abnormalities or adverse effects were observed in any of the rats upon
necropsy.

An initial pulmonary rat study which resulted in lethality in a
significant number of animals treated with either the live Aspergillus
flavus AF36 in Tween 80 or heat killed Aspergillus flavus AF36 in Tween
80. Onset of symptoms was rapid after dosing with all deaths occurring
by day four of the study. All rats surviving to day four of the study
recovered and all rats sacrificed (as scheduled) on day 8 or day 15 of
the study had totally eliminated viable Aspergillus flavus AF36 from the
lungs, caecal contents, and faeces. There was no evidence of
infectivity. The aetiology of deaths was unclear.  It appeared that
Aspergillus flavus AF36 prepared using Tween 80 caused a severe acute
inflammatory response. Retrospective literature review and consultation
with a toxicologist supported the theory that the responses were a
result of a synergism with Tween 80 and/or of Tween 80 breakdown
products formed during preparation of the spore suspension.

A second rat study was therefore undertaken. In the second study the
conidia were both washed from the wheat and suspended in sterile
physiological saline instead of Tween 80. Animals (2 male and 2 female
for each treatment level) were dosed at 0, 105, 106, 107, and 108 colony
forming units per rat. There were no clinical signs in any of the
treatment groups considered to be associated with the test substance. 
Rats were sacrificed at day 8 without treatment associated mortality. No
abnormalities were observed in any of the animals at the macroscopic
examination at termination.

Based on these two mammalian studies, it was concluded that Aspergillus
flavus AF36 does not present either a toxicological or an infectious
risk to mammals.

Data waivers were requested for the toxicology studies listed below on
the following basis:  (l) researchers and other workers have worked with
Aspergillus flavus AF36 at the Southern Regional Research Center for
over 10 years and in commercial fields (1996 to 1998) and in hand-picked
field plots (1989 to 1994) without report of any adverse health effects;
(2) Aspergillus flavus AF36 is widely distributed in the environment and
its occurrence is natural; and 3) the label will require applicators and
other handlers to wear Personal Protective Equipment (PPE) such as
waterproof gloves, a dust/mist filtering respirator with the appropriate
NIOSH approval prefix N-95, P-95, or R- 95, coveralls, long sleeved
shirt and long pants, and shoes plus socks, and goggles, so exposure
should not be a problem. Applications will involve aerial application by
mixers/handlers who are licensed and trained to even handle restricted
materials.  At the 10 lb/acre application rate of the formulated
material, the total amount of active ingredient is less than 0.01
lb/acre.  Applications of AF-36 do not significantly impact the total
amount of Aspergillus flavus in the soil or crop, but only change the
proportion of the AF-36 strain in relation to the overall soil or crop
population.  Since the product is applied as a granular formulation on
wheat or sorghum, exposure from drift is minimal.

Acute Dermal toxicity.  This study was waived based upon the lack of
toxicity in animals dosed orally. While Aspergillus flavus has been
reported to be a dermal sensitizer, this testing is not warranted, since
the PPE required on the label will provide adequate protection from
dermal exposure to workers and pesticide handlers.

Primary Dermal Irritation.  This study was waived based upon the lack of
toxicity in animals dosed orally.  Aerial applications will be made by
mixers/handlers and applicators, who are required to wear appropriate
PPE, and are licensed and trained to even handle restricted materials. 
At the 10 lb/acre application rate of the formulated material, the total
amount of active ingredient is less than 0.01 lb/acre.  Applications of
AF-36 do not significantly impact the total amount of Aspergillus flavus
in the soil or crop, but only change the proportion of the AF-36 strain
in relation to the overall population. Since the product is applied as a
granular formulation on wheat, exposure from drift is minimal.

4. Primary eye irritation.  This study was waived based upon the use of
appropriate PPE, including goggles, and other factors listed above.
Since the product is applied aerially as a granular formulation on wheat
or sorghum, exposure from drift is expected to be minimal.

5. Acute Intraperitoneal.  This study was waived based upon the lack of
toxicity in animals dosed orally and by intratracheal instillation.  In
addition, the factors listed above will mitigate exposure, which is
expected to be minimal.

Genotoxicity, reproductive and developmental toxicity, subchronic
toxicity and chronic toxicity.  These were not performed, since no
adverse effects were observed in the acute toxicology study Tier 1
studies. Tier II (885.3550), subchronic toxicology study (USEPA OPPTS
885.3600) and chronic feeding studies (guideline 152-50) are only
required if triggered by adverse effects observed in Tier I studies.

D. Aggregate Exposure

	1. Dietary exposure.

	i. Food.   Aspergillus flavus AF36 is a naturally occurring organism,
which does not produce aflatoxin and is thus safer than the
aflatoxin-producing Aspergillus flavus isolates that produce aflatoxin. 
Proposed uses and application rates will not result in increases in the
total population of A. flavus on the mature crop beyond naturally
occurring background levels.  Aspergillus flavus AF36, when applied to
the soil just prior to bloom, has been shown to significantly reduce the
levels of aflatoxin in pistachios at harvest without changing the
quantity of A. flavus on the crop.  Furthermore, the proposed use and
application rate will not increase exposure of humans to A. flavus by
dietary means.   Applications of Aspergillus flavus AF36 do not increase
the indigenous populations of A. flavus associated with the harvested
crop.  The applications merely alter the composition of the fungal
community associated with the mature crop so that aflatoxin producing
strains are far less frequent.  The result is a much lower incidence of
aflatoxins in the crop and in the environment associated with the
developing and mature crop.

	ii. Drinking water.  Aspergillus flavus AF36 is a naturally occurring
organism that is already widespread in the environment and is not
considered to be a risk to drinking water.  Both percolation through
soil and municipal treatment of drinking water would reduce the
possibility of exposure of Aspergillus flavus through the drinking
water.  Applications of Aspergillus flavus AF36 do not increase the
long-term populations of Aspergillus flavus in the environment, and thus
are not expected to influence the relationship of Aspergillus flavus to
water sources.  Applications merely change the composition of the
Aspergillus flavus community so that aflatoxins are less common in the
environment.

	2. Non-dietary exposure.  The potential for non-occupational,
non-dietary exposure to the general population is not expected to be
significant and is not expected to present any risk of adverse health
effects.

E. Cumulative Effects

There are no other registered products containing Aspergillus flavus
AF36 or any other isolates (strains) of the microbial active ingredient.
 Data submitted show that the fungal metabolite of concern which is
aflatoxin is not produced by Aspergillus flavus AF36 in the crop or in
artificial media in the lab.  Aspergillus flavus AF36 has been shown to
exclude aflatoxin producing fungi competitively from the developing crop
and to reduce aflatoxin contamination of almonds and figs.  Data show
that the proposed use will not result in appreciable increases in the
long term population of Aspergillus flavus on the crop beyond naturally
occurring levels.  Furthermore, there is no expectation of cumulative
effects with other pesticides.

 

F. Safety Determination

U.S. population.  Aspergillus flavus AF36 is a naturally occurring
organism.  This isolate has low toxicity as demonstrated by the acute
oral toxicity study in rats.  Studies have shown that treatment of
cotton fields just prior to flowering with sterile wheat seed colonized
by Aspergillus flavus AF36 at 10 lbs. per acre does not increase the
long-term populations of Aspergillus flavus either on the crop at
maturity or in the soil 1 year after application.  Based on this
information, IR-4 is of the opinion that the aggregate exposure to
Aspergillus flavus over a lifetime should not change with application of
Aspergillus flavus AF36, and exposure to both aflatoxin producing
Aspergillus flavus strains and aflatoxin should decrease.  This should
be beneficial to human health.  Thus, there is a reasonable certainty
that no harm will result from aggregate exposure to Aspergillus flavus
AF36.

2. Infants and children.  Based on the lack of toxicity and natural
occurrence, there is reasonable certainty that no harm to infants,
children, or adults will result from aggregate exposure to Aspergillus
flavus AF36.  Exempting Aspergillus flavus AF36 from the requirement of
a tolerance should pose no significant risk to humans or the
environment.

G. Effects on the Immune and Endocrine Systems

Aspergillus jlavus AF36 is a naturally occurring organism, which does
not produce aflatoxin, and is thus safer than the A. flavus isolates
that produce aflatoxin.  To date there is no evidence to suggest that
Aspergillus jlavus AF36 functions in a manner similar to any known
hormone, or that it acts as an endocrine disrupter.

H. Existing Tolerances

There is an existing exemption from the requirement of a tolerance for
residues of the microbial pesticide Aspergillus flavus AF36 in or on
cotton, gin byproducts; cotton, hulls; cotton, meal; cotton, refined
oil; cotton, undelinted seed.  

There is an existing exemption from the requirement of a tolerance for
residues of .Aspergillus flavus AF36 in or on corn, field, forage; corn,
field, grain; corn, field, stover; corn, field, aspirated grain
fractions; corn, sweet, kernel plus cob with husk removed; corn, sweet,
forage; corn, sweet, stover; corn, pop, grain; and corn, pop, stover,
when applied/used as an antifungal agent.  

There is an existing exemption from the requirement of a tolerance for
residues of Aspergillus flavus AF36 in or on pistachio when applied as
an antifungal agent and used in accordance with good agricultural
practices.  

There is also a time-limited exemption from the requirement of a
tolerance established for residues of Aspergillus flavus AF36, in or on
dried figs, resulting from use of the pesticide pursuant to a FIFRA
section 18 emergency exemption.  This time-limited exemption from the
requirement of a tolerance for residues of Aspergillus flavus AF36 in or
on dried figs will expire on December 31, 2017.

I. International Tolerances

There are no Codex maximum residue levels established for residues of
Aspergillus flavus AF36.  Aspergillus flavus AF36 containing products
are presently not registered for pest control outside of the United
States.

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