**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**  **- 8**  **05 9** 

**SUBMISSION: E**  **- mail the completed template to:**  **nesci.kimberly @epa.gov.**  **TEMPLATE: Andermatt Biocontrol AG**  **PP 3F**  **8221** EPA has received a pesticide petition **( 3F**  **8221 )**  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 tol erance for **Bacillus amyloliquefaciens**  **strain FZB 42**  **(Options (pick one)** 1. microbial pesticide for **Bacillus am**  **yloliquefaciens 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** 2

A. Product Name and Proposed Use Practices **RhizoVital 42 containing**  **Bacillus am**  **yloliquefaciens strain FZB42**  **is being submitted to EPA for registration.**  **It will be applied for preventive use against ma** **jor soil -**  **borne diseases on most crops. Applicatio n 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 **Fa mily:**  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 naturally** **** **infected 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 tox icity of the active ingredient**  **(see Toxicolog ical**  **Profile). O PPTS**  **Guideline 885.2000, section f, Background for**  **Residue Analysis of Mi crobial Pest Control Agents (MPC**  **A), states: “The Agenc** **y evaluates residue data for MPC As 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 excep**  **tion 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**  **am yloliquefaciens**  **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 Bioc**  **ontrol AG is**  **, therefore, submitting a petition to**  **establish an exemption from the requirement of a tolerance and an analytical** 3

**method is not required.** C. Mammalian Toxicological Profile **The full reports that support this petition and which are summ arized 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 5.7x10**  **9 CFU/mL of**  **B .**  **am**

**yloliquefaciens strain FZB42. Interim sacrifices were made on**  **three animals per sex on Days 3, 7, and 14; the remaining animals wer e 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 Day**  **s 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 t**  **o 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 10 3**  **7.1 x 10 4**  **7.9 x 10 2**  **5.1 x 10 2**  **5.6 x 10 1** 

**Urine samples CFU/ g**  **1 hr 24 hr**  **48 hr 72 hr**  **Day 14 6.1 x 10**  **2 0**  **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** **. amyloliquef**  **aciens Acute Dermal Toxicity:**  **Ten animals were dosed at 5,000 mg/kg of RhizoVital 42 containing 4.1x10 10**  **CFU/g of**  **B .**  **am yloliquefaciens**  **strain FZB42. No skin reactions or signs of toxicity were observed. All animals appeared healthy at necropsy. The LD50**  **was determined to**4

**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 (M P**  **C A** **) Treated animals were dosed with**  **1.1x10 9**  **CFU/rat of RhizoVital 42**  **. Interim sacrifices were made on three animals per sex on Days 0 (3 hours post** **- application),**  **3, 7, and 1 4; 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**  **nod es**  **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. amyloliquefac** **iens 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**  **, excep t 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 (4.1x10 10**  **CFU/g) of RhizoVital 4 2. No**  **irrita tion or**  **other clinical signs were observed. RhizoVital 4**  **2 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 contai ning 4.1x10**  **10 CFU/g**  **of B. amyloliquefacien**  **s 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.** 5

**Hypersensitivity Incidents: No reports of hypersensitivity have been associated with**  **Bacillus amyloliquefaciens strain FZB42.**  **Acute Intravenous**  **T oxicity/** **P athogenicity:**  **Twenty -**  **five male and 25 female Spr a**  **gue Dawley rats were assigned as follows: 5**  **animals/sex -**  **Group I, Untreated**  **5 animals/sex**  **- Group II, Inactive M**  **P C**  **A 15**  **animals/sex -**  **Group III, M P**  **C A**  **Group II an d Group III animals were dosed with**  **0.1 mL of RhizoVital 42**  **intravenously via tail vein. The suspension**  **was prepared so as**  **to contain 3.0x**  **10 9**  **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**  **grou p.**  **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, a**  **ll samples were**  **negative for B. amyloliquefacien**  **s 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 show**  **ed a pattern of clearance. Mesenteric**  **l ym** **ph nodes: By Day 21, the test substance sho** **wed 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, dieta ry, 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**  **amyloliquefa ciens**  **strain FZB42 should not be of concern due to the low toxicity shown across the entire toxicological profile** 6

**(see above).** ii. Drinking water

. **Bacillus amyloliquefaciens is not likely to be present in**  **aquatic environments at significant levels,**  **sin ce 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 expo**  **sure. 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 stra**  **in 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 wor** **ld’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 info**  **rmation 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 a**  **re 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 suppressi** **ve action. This passive mode of action does not have a common chemical** **** **based mechanism of toxicity as can sometimes be found with typical organic chemicals.**  F . Safety Determination 1. U.S. population. **Bacillus am**  **yloliquefaciens strain FZB42 exhibits**  **low ma**  **mmalian toxicity /** **pathogenicity ,**  **as demonstrated by the submitted**  **toxicolog y**  **studies and summarized in the toxicological ical profile. When**  **Bacillus amyloliquefaciens**  **strain FZB42 is used on raw agricultural commodities in accordance with good agricu**  **ltural 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 c**  **rops 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 .** 7

G. Effects on the Immune and Endocrin e 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**  **.**