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

SUBMISSION: E-mail the completed template to: nesci.kimberly@epa.gov.

TEMPLATE:

Andermatt Biocontrol AG

PP3F8221

EPA has received a pesticide petition (3F8221) from Andermatt Biocontrol AG, 

Stahlermatten, 6 CH-6146, Grossdietwil, Switzerland 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.

(Options (pick one)

2. to establish an exemption from the requirement of a tolerance for Bacillus 

amyloliquefaciens strain FZB42

(Options (pick one)

1. microbial pesticide for Bacillus amyloliquefaciens strain FZB42

Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, Andermatt Biocontrol AG

has submitted the following summary of information, data, and arguments in support of 

their pesticide petition. This summary was prepared by SciReg, Inc. 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. Andermatt Biocontrol AG Petition Summary

3F8221
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A. Product Name and Proposed Use Practices

RhizoVital 42 containing Bacillus amyloliquefaciens strain FZB42 is being 

submitted to EPA for registration. It will be applied for preventive use against 

major soil-borne diseases on most crops. Application rates are projected to be up to 

28 fl. oz./acre/application; up to three applications per season are anticipated

B. Product Identity/Chemistry

1. Identity of the pesticide and corresponding residues.

Kingdom: Bacteria

Phylum: Firmicutes

Class: Bacilli

Order: Bacillales

Family: Bacillaceae

Genus: Bacillus

Species: B. amyloliquefaciens (Gram-positive)

Bacillus amyloliquefaciens is a naturally-occurring, ubiquitous soil microbe. 

Bacillus amyloliquefaciens strain FZB42 was isolated in Germany from naturallyinfected soil. It is not genetically modified. Bacillus amyloliquefaciens strain FZB42 

is intended for preventive use against major soil-borne diseases on a variety of 

crops, ornamental plants, and turf.

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

residue. No magnitude of residue (MOR) studies have been conducted with Bacillus 

amyloliquefaciens strain FZB42 based on the low toxicity of the active ingredient

(see Toxicological Profile). OPPTS Guideline 885.2000, section f, Background for 

Residue Analysis of Microbial Pest Control Agents (MPCA), states: 

“The Agency evaluates residue data for MPCAs used on food, feed, or raw 

agricultural commodities only if toxic or other harmful properties were observed in 

the maximum hazard toxicology tests (Tier I) prescribed in toxicology test 

guidelines OPPTS 885.3050 through 885.3650 of this series. If Tier I toxicology tests 

indicate no toxic or other harmful properties, then no residue data (with the general 

exception of a monitoring method) would be indicated and thus a recommendation 

for an exemption from the requirements of a tolerance can be made.”

3. A statement of why an analytical method of detecting and measuring the levels 

of the pesticide residue are not needed. While Bacillus amyloliquefaciens strain FZB42 

residues may be present on treated food/feed commodities, based on the submitted 

toxicology data, Bacillus amyloliquefaciens strain FZB42 is virtually non-toxic and is 

not pathogenic. Andermatt Biocontrol AG is, therefore, submitting a petition to 

establish an exemption from the requirement of a tolerance and an analytical 
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method is not required.

C. Mammalian Toxicological Profile

The full reports that support this petition and which are summarized in this 

section were previously submitted to the Agency by Andermatt Biocontrol AG in 

support of registration. Based on the data submitted, it is established that Bacillus 

amyloliquefaciens strain FZB42 is neither toxic, nor pathogenic to humans. 

Acute Oral Toxicity/Pathogenicity: 

Twenty-four male and 24 female Sprague-Dawley rats were dosed at 5.7x109

CFU/mL of B. amyloliquefaciens strain FZB42. Interim sacrifices were made on 

three animals per sex on Days 3, 7, and 14; the remaining animals were sacrificed on 

Day 21, the termination of the study. There were no deaths and all animals 

appeared normal throughout the entire study. 

Feces and urine samples were collected at 1, 24, 48 and 72 hours and Day 14. Blood, 

brain, lung, liver, kidney, spleen, lymph nodes, and cecum content were evaluated 

on Days 3, 7, 14, and 21 (all other tissues) for test article-treated animals, untreated 

control animals, and inactivated test article-treated animals). By 24 hours after 

dosing, the test substance appeared to have cleared from the urine. By Day 14, the 

test substance showed a pattern of clearance from the feces.

A mycological summary of these evaluations follows.

Feces samples CFU/g

1 hr 24 hr 48 hr 72 hr Day 14 

1.1 x 1037.1 x 1047.9 x 1025.1 x 1025.6 x 101

Urine samples CFU/g

1 hr 24 hr 48 hr 72 hr Day 14 

6.1 x 1020 0 0 0

Blood: All samples negative for B. amyloliquefaciens

Brain: All samples negative for B. amyloliquefaciens

Lung: All samples negative for B. amyloliquefaciens

Liver: All samples negative for B. amyloliquefaciens

Kidney: All samples negative for B. amyloliquefaciens

Spleen: All samples negative for B. amyloliquefaciens

Lymph nodes: All samples negative for B. amyloliquefaciens

Cecum: All samples negative for B. amyloliquefaciens

Acute Dermal Toxicity: 

Ten animals were dosed at 5,000 mg/kg of RhizoVital 42 containing 4.1x1010 CFU/g 

of B. amyloliquefaciens strain FZB42. No skin reactions or signs of toxicity were 

observed. All animals appeared healthy at necropsy. The LD50 was determined to 
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be greater than 5,000 mg/kg.

Acute Pulmonary Toxicity/Pathogenicity: 

Thirty-two male and 32 female Sprague-Dawley rats were assigned as follows:

5 animals/sex - Group I, Untreated

4 animals/sex - Group II, Shelf Control

5 animals/sex - Group III, Inactive

18 animals/sex – Group IV, Microbial Pest Control Agent (MPCA)

Treated animals were dosed with 1.1x109 CFU/rat of RhizoVital 42. Interim 

sacrifices were made on three animals per sex on Days 0 (3 hours post-application), 

3, 7, and 14; the remaining animals were sacrificed on Day 21, the termination of the 

study. 

There were no deaths and there were no clinical signs noted in any animal of any 

group.

Blood, brain, lung, liver, kidney, spleen, cecum contents, and mesenteric lymph 

nodes were evaluated on Day 0 (feces only) and Day 3, 7, 14, and 21 (all other 

tissues) for all animals. A mycological summary of these evaluations follows.

Blood: All samples negative for B. amyloliquefaciens

Brain: All samples negative for B. amyloliquefaciens

Lung: All samples negative for B. amyloliquefaciens, except for one female on Day 3

Liver: All samples negative for B. amyloliquefaciens

Kidney: All samples negative for B. amyloliquefaciens

Spleen: All samples negative for B. amyloliquefaciens, except for one male on Day 3

Mesenteric Lymph nodes: All samples negative for B. amyloliquefaciens

Cecum contents: All samples negative for B. amyloliquefaciens

Eye Irritation: 

Three rabbits were dosed with 0.1 mL (4.1x1010 CFU/g) of RhizoVital 42. No

irritation or other clinical signs were observed. RhizoVital 42 is considered to be 

non-irritating.

Dermal Irritation: 

Three rabbits were each dosed with a semi-occlusive liquid patch of 0.5 mL of 

RhizoVital 42 for 4 hours. The test substance contained containing 4.1x1010 CFU/g

of B. amyloliquefaciens strain FZB42. Grade 1 erythema was observed in all animals 

60 minutes after patch removal. No clinical signs were observed 2 hours after patch 

removal. RhizoVital 42 is considered to be virtually non-irritating.
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Hypersensitivity Incidents: 

No reports of hypersensitivity have been associated with Bacillus amyloliquefaciens

strain FZB42.

Acute Intravenous Toxicity/Pathogenicity:

Twenty-five male and 25 female Sprague Dawley rats were assigned as follows: 

5 animals/sex - Group I, Untreated

5 animals/sex - Group II, Inactive MPCA

15 animals/sex - Group III, MPCA

Group II and Group III animals were dosed with 0.1 mL of RhizoVital 42

intravenously via tail vein. The suspension was prepared so as to contain 3.0x109

CFU/rat. Interim sacrifices were made on three animals per sex on Days 3, 7, 14 and 

21; the remaining animals were sacrificed on Day 28, the termination of the study. 

There were no deaths and there were no clinical signs noted in any animal of any 

group.

Blood, brain, lung, liver, kidney, spleen, cecum contents, and mesenteric lymph 

nodes were evaluated on Day 0 (blood only) and Day 3, 7, 14, and 21 (all other 

tissues) for MPCA treated animals. A mycological summary of these evaluations 

follows.

Blood: By Day 3, the test substance showed a pattern of clearance. All samples negative 

for B. amyloliquefaciens on Day 21.

Brain: By Day 14, the test substance showed a pattern of clearance. All samples negative 

for B. amyloliquefaciens on Day 21.

Lung: On Day 21, all samples were negative for B. amyloliquefaciens except for one 

male.

Liver: By Day 21, the test substance showed a pattern of clearance.

Kidney: By Day 21, the test substance showed a pattern of clearance.

Spleen: By Day 21, the test substance showed a pattern of clearance.

Mesenteric lymph nodes: By Day 21, the test substance showed a pattern of clearance.

Cecum contents: By Day 21, the test substance showed a pattern of clearance.

D. Aggregate Exposure

The potential for non-occupational, dietary, and non-dietary exposure to Bacillus

amyloliquefaciens strain FZB42 by the general population, including infants and 

children, is likely through ingestion of treated crops. 

1. Dietary exposure.

i. Food. Dietary exposure to Bacillus amyloliquefaciens strain FZB42 should 

not be of concern due to the low toxicity shown across the entire toxicological profile 
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(see above).

ii. Drinking water. Bacillus amyloliquefaciens is not likely to be present in 

aquatic environments at significant levels, since it is not known to inhabit such 

environments. Anticipated agricultural practice prohibits direct application of the 

end-use product directly to water and natural environmental filtration processes, 

such as soil and vegetation, further limit aquatic exposure. Additionally, the label 

requires the product to be immediately watered in after application, so that the 

microbes are able reach the root zones of the target plants.

2. Non-dietary exposure. Non-dietary exposure to Bacillus amyloliquefaciens

strain FZB42 will be much more likely through handling by formulators and 

applicators, but in light of the subject strain’s toxicological profile, should be of little 

concern. Although the organism is ubiquitous, levels of exposure in broad segments 

of the world’s population has not resulted in any significant reported adverse 

effects.

E. Cumulative Effects

Section 408(b)(2) of the FFDCA requires that, when considering whether to 

establish an exemption from a tolerance, the Agency consider available information 

concerning the cumulative effects of the particular ingredient’s residues and other 

substances that have a common mechanism of toxicity. These considerations include 

the possible cumulative effects of such residues on infants and children. Bacillus

species are naturally-occurring, ubiquitous soil microbes. Bacillus amyloliquefaciens

strain FZB42 is neither toxic, nor pathogenic. The end-use product, RhizoVital 42, 

is used preventively against major soil-borne diseases on most crops through 

suppressive action. This passive mode of action does not have a common chemicalbased mechanism of toxicity as can sometimes be found with typical organic 

chemicals.

F. Safety Determination

1. U.S. population. Bacillus amyloliquefaciens strain FZB42 exhibits low 

mammalian toxicity/pathogenicity, as demonstrated by the submitted toxicology 

studies and summarized in the toxicological ical profile. When Bacillus 

amyloliquefaciens strain FZB42 is used on raw agricultural commodities in 

accordance with good agricultural practice, it meets the reasonable certainty of no 

harm requirement.

2. Infants and children. It is believed that the potential exposure of infants and 

children to residues of Bacillus amyloliquefaciens strain FZB42 through their use to 

protect crops from diseases is no greater than that of the general population. 

Available information does not suggest that infants and children are more 

susceptible to Bacillus amyloliquefaciens strain FZB42 than are adults.
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G. Effects on the Immune and Endocrine Systems

Extensive literature searches and review of the Bacillus amyloliquefaciens

strain FZB42 toxicity/pathogenicity data indicate there is no evidence that this 

strain is an endocrine disruptor or that it has any known effect on immune systems.

H. Existing Tolerances

There are no known agricultural tolerances or tolerance exemptions for 

Bacillus amyloliquefaciens strain FZB42.

I. International Tolerances

There are no known international tolerances for Bacillus amyloliquefaciens

strain FZB42.