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: [Kimberly Nesci ; 

(703) 308-8059]

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:

[BASF Corporation]

[Petition No. 4F8336]

EPA has received a pesticide petition ([No. 4F8336]) from [BASF Corporation], 

[26 Davis Drive, Research Triangle Park, NC 27709] 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 amendment/expansion of an existing tolerance 

exemption for the microbial pesticide [Bacillus amyloliquefaciens MBI 600

(antecedent Bacillus subtilis MBI 600)]

Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, [BASF 

Corporation] has submitted the following summary of information, data, and arguments 

in support of their pesticide petition. This summary was prepared by [BASF 

Corporation] 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. [BASF Corporation] Petition Summary

[Petition No. 4F8336 to AMEND 40 CFR § 180.1128 Bacillus subtilis MBI

600; exemption from the requirement of a tolerance; to change the species 

name to “amyloliquefaciens” and revise the exemption to state as follows:
2

“40 CFR § 180.1128 Bacillus amyloliquefaciens MBI 600; exemption 

from the requirement of a tolerance.

An exemption from the requirement of a tolerance is established for 

residues of the biofungicide Bacillus amyloliquefaciens MBI 600 

(antecedent Bacillus subtilis MBI 600) in or on all food commodities, 

including residues resulting from post-harvest uses, when applied or 

used in accordance with good agricultural practices.”]

A. Product Name and Proposed Use Practices

[Following is a list of all the currently EPA registered pesticide products that 

will be affected by the change in the species name from subtilis to amyloliquefaciens.

EPA Registration No. Product Name*

71840-1 Subtilex® MUP

71840-2 HiStick® NT

71840-5 Integral® Liquid Biological Fungicide

71840-6 Integral® II Liquid Biofungicide

71840-8 Subtilex® NG Biofungicide

71840-9 Bacillus subtilis strain MBI 600 Technical

* Serifel Biofungicide (EPA Registration No. 71840-18 is already registered with the 

amyloliquefaciens species designation.

The end-use products containing the MBI 600 Integral®, Integral® II, HiStick® 

NT Subtilex® NG, SUBTILEX® MUP, and Bacillus subtilis strain MBI 600

Technical biological fungicide products and are currently used to suppress disease 

organisms such as Botrytis, Fusarium, Rhizoctonia, and Alternaria as well as those 

organisms causing powdery mildew and anthracnose. 

Current use sites include commercial application for seed treatment, foliar, infurrow, injected and soil drench applications for greenhouse and field crops and 

ornamentals.

The use patterns and application rates of Bacillus amyloliquefaciens MBI 600

(antecedent Bacillus subtilis MBI 600) minimize the potential for exposure above the 

natural background levels. 

B. Product Identity/Chemistry
3

1. Identity of the pesticide and corresponding residues. [Bacillus 

amyloliquefaciens MBI 600 (antecedent Bacillus subtilis MBI 600); CAS 

No. 68083-70-0. When used as directed, Bacillus amyloliquefaciens MBI 

600 is not expected to result in residues that are of toxicological concern.

Bacillus amyloliquefaciens is a naturally occurring soil saprophyte that is 

found in soils worldwide. Strains of this organism are generally not 

regarded as human or animal pathogens. Bacillus amyloliquefaciens 

MBI 600 (antecedent Bacillus subtilis MBI 600) was first isolated from 

faba bean plants growing at Nottingham University School of 

Agriculture in Sutton Boningham, UK. This strain was subsequently 

researched, developed, and commercialized by Gustafson, LLC (first 

registered as EPIC Biological Fungicide in 1994). The registration 

transferred to MicroBio in 1998 and transferred again to Becker 

Underwood following their acquisition of MicroBio in 2000. 

In 2013, BASF Corporation acquired Becker Underwood and currently 

owns all Becker Underwood pesticide product registrations and 

associated data submitted in support of those registrations.]

2. Magnitude of residues at the time of harvest and method used to determine the 

residue. [An analytical method for residues is not applicable.]

3. 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 of Bacillus

amyloliquefaciens MBI 600 (antecedent Bacillus subtilis MBI 600) is not necessary as

Bacillus amyloliquefaciens MBI 600 (antecedent Bacillus subtilis MBI 600) has an

exemption from the requirement of a tolerance without numerical limitations.]

C. Mammalian Toxicological Profile

[Toxicology and pathogenicity studies on Bacillus amyloliquefaciens MBI 600

(antecedent Bacillus subtilis MBI 600) were previously accepted by the EPA to 

support the current exemption from the requirement of a tolerance for residues of 

MBI 600. The data consist of laboratory animal (in vivo) toxicity studies conducted 

in accordance with Good Laboratory Practice.

1. Acute Oral Toxicity/Pathogenicity: (MRID 419074-02) Bacillus 

amyloliquefaciens MBI 600 was not pathogenic, infective or toxic in rats 

when orally dosed with 2 x 108 CFU per animal. Toxicity category IV.

2. Acute Dermal Toxicity: (MRID 419074-03) Bacillus amyloliquefaciens MBI 

600 was not toxic for rats when a single 5 x 1010 CFU/animal (2 ml/kg body 

weight) dose was administered dermally to rabbits. Toxicity category IV.

3. Acute Pulmonary Toxicity/Pathogenicity: (MRID 419074-04 Bacillus 

amyloliquefaciens MBI 600 was not infective, pathogenic or toxic to rats 

dosed intratracheally with 3.4 x 108 CFU of the test material. Toxicity 

category IV. 
4

4. Acute Intravenous Toxicity/Pathogenicity: (MRID 419074-05) Bacillus 

amyloliquefaciens MBI 600 was not infective, pathogenic or toxic for rats 

when dosed intravenously with approximately 4 x 107 CFU of the test 

material. 

5. Primary Eye Irritation: (MRID 419074-06) Bacillus amyloliquefaciens MBI 

600 produced a slight irritation when a single 0.1 g ocular does was 

administered to rabbits. Occular irritation dissipated 4 days post dosing. 

Toxicity category IV.

6. Hypersensitivity Studies: (MRID 419074-07, April 1989) An overall 

moderated skin sensitization reaction was noted in guinea pigs 24 to 72 hours 

after treatment with Bacillus amyloliquefaciens MBI 600. (MRID 462059-01, 

December 2003) No irritation was observe after the challenge with 

SUBTILEX® Biological Fungicide indicating the product does not elicit a 

sensitizing reaction in guinea pigs and does not appear to be a dermal 

sensitizer.

7. Hypersensitivity incidents: To date, there are no reports of incidents of 

human health or ecological hazards from exposure to Bacillus 

amyloliquefaciens MBI 600 .

Results of these studies indicate the Bacillus amyloliquefaciens MBI 600 (antecedent 

Bacillus subtilis MBI 600) is not toxic to test animals when administered via the oral, 

dermal, pulmonary or intravenous routes of exposure. Additionally, it is not 

infective or pathogenic when administered via the oral, pulmonary or intravenous 

routes. Because of the lack of toxicity, pathogenicity, and infectivity, the EPA 

concluded the product should not pose a risk to wild mammalian species. 

No reports of hypersensitivity have been recorded from personnel working with this 

organism. MBI 600 does not produce significant quantities of extracellular enzymes 

or possess other virulence factors that would predispose it to cause infection. 

Based on the studies and other available information submitted for Bacillus 

amyloliquefaciens MBI 600 (antecedent Bacillus subtilis MBI 600), the EPA found 

no evidence of risks to human health.]

D. Aggregate Exposure

1. Dietary exposure. []

i. Food. [The proposed use pattern may result in dietary exposure with 

possible residues in or on agricultural commodities. However, little to no risk 

is expected for the general population, including infants and children, or 

animals because of the lack of demonstrated toxicity/pathogenicity of 

Bacillus amyloliquefaciens MBI 600. Furthermore, Bacillus amyloliquefaciens

is used in the production of fermented foods and commonly consumed by 

humans with no adverse health effects. Additionally, Bacillus 

amyloliquefaciens is commonly used on a large, industrial scale basis to 

produce food grade products.
5

Acute dietary assessments were conducted on October 10, 1993 in a Science 

Advisory Board (SAB) review for the registration of a Bacillus subtilis 

(amyloliquefaciens) MBI 600 concentrate product. No toxicological end 

points of concern were identified. There are no dietary risks that exceed the 

Agency’s Level of Concern (LOC).]

ii. Drinking water. [The potential for transfer of Bacillus amyloliquefaciens

MBI 600 to surface or ground water during run-off associated with intended 

use applications is considered minimal to non-existent, due to its percolation 

through, and resulting capture in soil. Accordingly, the use of this microbial 

pest control agent on terrestrial plants is not anticipated to negatively impact 

the quality of drinking water.]

2. Non-dietary exposure. [The potential for dermal exposure to pesticide residues of 

Bacillus amyloliquefaciens MBI 600 for the general population, including infants 

and children, is unlikely as the use sites are limited to agricultural and horticultural 

applications. In addition, as the microbe is a ubiquitous, naturally occurring soil 

bacterium, it is likely many individuals have had prior exposure. Accordingly, any 

potential increase in dermal exposure due to pesticidal use of Bacillus 

amyloliquefaciens MBI 600 would be inconsequential. Furthermore, as summarized 

previously, MBI 600 is of low dermal toxicity - Toxicity Category IV. 

Similarly, the inhalation exposure to pesticide residues of Bacillus amyloliquefaciens

MBI 600 for the general population, including infants and children is unlikely 

because potential use sites are agricultural and horticultural. Also, as stated before, 

the microbe is a ubiquitous, naturally occurring soil bacterium, and it is likely many 

individuals have had prior exposure. The acute pulmonary toxicity/pathogenicity 

testing of Bacillus amyloliquefaciens MBI 600 demonstrated it is neither toxic, 

pathogenic nor infective - Toxicity Category IV Therefore, any potential increase in 

inhalation exposure due to pesticidal use of Bacillus amyloliquefaciens MBI 600 

would be inconsequential. 

The results of toxicity testing indicate there is no risk to human health or the 

environment from Bacillus amyloliquefaciens MBI 600. It is not toxic to test animals 

when administered via the oral, dermal, intravenous or pulmonary routes of 

exposure. Nor is it infective or pathogenic to test animals when administered via the 

oral, pulmonary and intravenous routes. In addition, no reports of hypersensitivity 

have been recorded from personnel working with this organism. 

MBI 600 does not produce recognized toxins, enzymes, or virulence factors 

normally associated with mammalian invasiveness or toxicity. The limited survival 

of MBI 600 and the lack of acute toxicity indicate that both the hazard and the 

exposure associated with the use of MBI 600 are low. Non-dietary exposures would 

not be expected to pose any quantifiable risk due to a lack of residues of 

toxicological concern.]
6

E. Cumulative Effects

[Bacillus amyloliquefaciens MBI 600 is non-toxic to mammals. Because no 

mechanism of pathogenicity or toxicity in mammals has been identified for this 

organism, no cumulative effects from the interaction of residues of this product with 

other related microbial pesticides are anticipated when this product is used as 

directed on the label and in accordance with good agricultural/horticultural 

practices.

It is not expected that, when used as directed, Bacillus amyloliquefaciens MBI 600 

would result in residues that are of toxicological concern. The intended use of 

Bacillus amyloliquefaciens MBI 600 is to apply it to growing media, seeds, and 

growing plants and crops for the purposes of disease control. Bacillus 

amyloliquefaciens is widely distributed in the US and around the world. Application 

of Bacillus amyloliquefaciens MBI 600 to seeds, foliage or soil will not result in any 

significant increases in overall concentrations of Bacillus amyloliquefaciens in the 

environment. 

The results of toxicity testing indicate there is no risk to human health or the 

environment from Bacillus amyloliquefaciens MBI 600. There are no reports of 

ecological or human health hazards caused by Bacillus amyloliquefaciens in general 

or the Bacillus amyloliquefaciens MBI 600 in specific. MBI 600 does not produce 

significant quantities of extracellular enzymes or possess other virulence factors that 

would predispose it to cause infection. The absence of acute toxicity or pathogenicity 

in laboratory animals demonstrates the benign nature of this .]

F. Safety Determination

1. U.S. population. [There is reasonable certainty that no harm to the US population, 

including infants and children, will result from aggregate exposure to residues of 

Bacillus amyloliquefaciens MBI 600 from its use as a microbial pest control agent. 

The results of toxicity testing indicate there is no risk to human health or the 

environment from Bacillus amyloliquefaciens MBI 600. It is not toxic to test animals 

when administered via the oral, dermal, intravenous or pulmonary routes of 

exposure. Nor is it infective or pathogenic to test animals when administered via the 

oral, pulmonary and intravenous routes. Bacillus amyloliquefaciens MBI 600 does 

not produce recognized toxins, enzymes, or virulence factors normally associated 

with mammalian invasiveness or toxicity. In addition, no reports of hypersensitivity 

have been recorded from personnel working with this organism.]

2. Infants and children. [For the same reasons stated previously, there is 

reasonable certainty that no harm to infants and children will result from aggregate 

exposure to residues of Bacillus amyloliquefaciens MBI 600 from its use as a 

microbial pest control agent.]
7

G. Effects on the Immune and Endocrine Systems

[There is no evidence to date suggesting Bacillus amyloliquefaciens MBI 600 

functions in a manner similar to any known hormone, or that it acts as an endocrine 

disrupter.]

H. Existing Tolerances

[40 CFR 180.1128 Bacillus subtilis MBI 600; exemption from the requirement of a 

tolerance.

“An exemption from the requirement of a tolerance is established for 

residues of the biofungicide Bacillus subtilis MBI 600 in or on all food 

commodities, including residues resulting from post-harvest uses, 

when applied or used in accordance with good agricultural 

practices.”]

I. International Tolerances

[A Codex Alimentarium Commission Maximum Residue Level (MRL) is not 

required for Bacillus amyloliquefaciens MBI 600.]