Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations for Bacillus mycoides isolate J

                    Docket ID Number: EPA-HQ-OPP-2014-0920
                               Date: May 9, 2016
                                       
Section 408(c)(2)(A)(i) of FFDCA allows the U.S. Environmental Protection Agency (EPA) to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the EPA determines that the exemption is "safe." Section 408(c)(2)(A)(ii) of FFDCA defines "safe" to mean that "there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information." This includes exposure through drinking water and in residential settings but does not include occupational exposure. Pursuant to FFDCA section 408(c)(2)(B), in establishing or maintaining in effect an exemption from the requirement of a tolerance, the EPA must take into account the factors set forth in FFDCA section 408(b)(2)(C), which require the EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance or tolerance exemption, and to "ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue . . . ." Additionally, FFDCA section 408(b)(2)(D) requires that the EPA consider "available information concerning the cumulative effects of [a particular pesticide's] . . . residues and other substances that have a common mechanism of toxicity."
 The EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, the EPA determines the toxicity of the pesticide. Second, the EPA examines exposure to the pesticide through food, drinking water, and other exposures that occur as a result of pesticide use in residential settings.
I.  Summary of Petitioned-for Tolerance Exemption
In the Federal Register of January 28, 2015 (80 FR 4525), the EPA issued a notice pursuant to FFDCA section 408(d)(3), 21 U.S.C. 346a(d)(3), announcing the filing of a pesticide tolerance petition (PP 4F8252) by Certis USA LLC, 9145 Guilford Rd., Suite 175, Columbia, MD 21046. The petition requested that 40 CFR part 180 be amended by establishing an exemption from the requirement of a tolerance for residues of Bacillus mycoides isolate J, in or on all agricultural commodities. That document referenced a summary of the petition prepared by the petitioner Certis USA LLC, which is available in Docket ID Number EPA-HQ-OPP-2014-0920 via http://www.regulations.gov.
II.  Toxicological Profile
Consistent with FFDCA section 408(b)(2)(D), the EPA reviewed the available scientific data and other relevant information on Bacillus mycoides isolate J, and considered its validity, completeness, and reliability, as well as the relationship of this information to human risk. The EPA also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.
 Overview of Bacillus mycoides isolate J
Bacillus mycoides isolate J is in the Bacillus cereus group of Gram positive microbes. Bacillus species are primarily soil and water associated and are commonly found in a variety of fresh produce and other foods with no ill effects from ingestion. Bacillus mycoides is very commonly found in soils, fresh water and all kinds of foods including those consumed with minimal processing. A literature review reveals that Bacillus mycoides has not been shown as the cause of any foodborne illnesses. 

Temporary exemptions from the requirement of a tolerance for Bacillus mycoides isolate J were previously approved to support two experimental use permits and a FIFRA section 18 emergency exemption. At this time, 40 CFR 180.1269 lists a temporary exemption that expired on December 31, 2015 for the FIFRA section 18 emergency exemption for potatoes and this paragraph will need to be removed.

The applicant claims that Bacillus mycoides isolate J acts a biological disease control agent that reduces the occurrence and severity of plant disease by triggering the plant's natural defense mechanisms against pathogens by a mechanism known as "Systemic Acquired Resistance" and that this microorganism does not directly affect plant pathogens.

The Bacillus mycoides isolate J manufacturing process was refined.  Subsequent testing after these changes did not result in any pharmacological or toxicological effects after intratracheal dosing, and resulted in no lethality using the current manufacturing process. Based on extensive animal testing of the Bacillus mycoides isolate J active ingredient no toxicity, irritation, infectivity, pathogenicity or other adverse effects from dietary exposure to Bacillus mycoides isolate J are expected. The overall conclusions from all toxicological information submitted by the petitioner are briefly described below, and more in-depth synopses of study results can be found in the Refer to risk assessments in docket number EPA-HQ-OPP-2015-0007, for additional information. (Ref. 1 and 2)



 Microbial Pesticide Toxicology Data Requirements
 Acute Pulmonary Toxicity and Pathogenicity - Rat. [OCSPP Guideline: 885.3150; (Master Record Identification Number MRID No.: 468448-01)]. An acceptable acute pulmonary toxicity and pathogenicity study demonstrated that Bacillus mycoides isolate J is not infective when instilled at 1.1 x 10[8] CFU/rat, though a pattern of clearance was not established. An explanation was not provided about the results of the preliminary assay where 6 rats that had been treated with unwashed spores, and 2 of 6 rats died within 26 hours.  (Ref. 1 and 3). Effects seen from the preliminary assay are likely attributable to excessive amounts of Bacillus enzymes in the lungs, which would not otherwise occur else from concentrated application of the pesticide.

 Waiver Rationale for Acute Inhalation Toxicity  -  Rat. [OCSPP Guideline: 870.1300 (MRID No.: 493324-07)] and Additional Confirmatory Testing for Acute Pulmonary Toxicity and Pathogenicity - Rat. [OCSPP Guideline: 885.3150 (MRID No.:498474-02).  The rationale for waiving the 14 day acute inhalation study submitted explained that Bacillus mycoides is widespread and likely ubiquitously found in all soil types, and in surface waters and that there have been no reported adverse effects to humans or animals via the inhalation route of exposure. Exposure on food products is common with no known health concerns, as demonstrated by a literature review. However the Agency finds that fermentation and production of a concentrated spore preparation is not assumed to be a natural exposure and so safety by inhalation cannot be inferred from such a literature review concerning natural exposures. Bacillus species in soil are known to produce copious amounts of degradative enzymes, such as proteases, useful for utilizing various food sources. Some of these have been produced industrially for products such as drain cleaners and laundry detergents. Likely the mortalities and lung lesions observed in the provided Pulmonary Toxicity and Pathogenicity study, and severe eye irritation noted in an Acute Eye Irritation study are the result of these enzymes. Subsequently the manufacturing process was refined and testing afterwards of the test substance with similar intratracheal dosing did not result in any pharmacological or toxicological effects nor any lethality.

An acceptable 48 hour acute pulmonary toxicity study was submitted. The 48-hour tracheal instillation lethal dose (LD50) which is a statistically derived single dose that can be expected to cause death in 50% of test animals for Bacillus mycoides isolate J TGAI is greater than 10[8] CFU/rat. There were no treatment related clinical signs or changes in body weight (EPA Toxicity Category III). (Ref. 1 and 2).
  
 Acute Injection Toxicity and Pathogenicity - Rat. [OCSPP Guideline: 885.3200; (MRID No.: 495815-06)]. An acceptable acute intravenous injection toxicity and pathogenicity study demonstrated that Bacillus mycoides isolate J is not toxic, infective, or pathogenic when instilled at 1 x 10[7] CFU/rat  -  a pattern of clearance was established. (Ref. 1 and 3).

 Acute Oral Toxicity  -  Rat. [OCSPP Guideline: 870.1100 (MRID No.: 493324-05)]. An acceptable acute oral toxicity study, demonstrated that BmJ WG (The end use product containing Bacillus mycoides isolate J) is not toxic, The oral median lethal dose (LD50), which is a statistically derived single dose that can be expected to cause death in 50% of test animals, was greater than 5,000 mg/Kg for female rats (EPA Toxicity Category IV). (Ref. 1 and 3).

 Acute Dermal Toxicity  -  Rat. [OCSPP Guideline: 870.1200 (MRID No.: 493324-06)] An acceptable acute dermal toxicity study demonstrated that BmJ WG was not toxic to male and female rats when dosed at 5,050 mg/kg of body weight for 24 hours. The dermal LD50 was greater than 5,050 mg/kg for female and male rats (EPA Toxicity Category IV). (Ref. 1 and 3).

 Primary Eye Irritation  -  Rabbit. [OCSPP Guideline: 870.2400 (MRID No.: 493324-08)] An acceptable primary eye irritation study demonstrated that BmJ WG was severely irritating to the eye.  Corneal opacity was noted in all rabbits immediately following exposure with clearance by 24 hours. Positive conjunctival irritation was noted on 3 of the 3 rabbits treated 24 hours after test material instillation with resolution by 48 hours (EPA Toxicity Category III). (Ref. 1and 3).

 Primary Dermal Irritation Study  -  Rabbit. [OCSPP Guideline: 870.2500 (MRID No.: 493324-09)] An acceptable primary dermal irritation study demonstrated that BmJ WG was non-irritating (EPA Toxicity Category IV). (Ref. 1 and 3).
 


III.  Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the EPA to consider available information concerning exposures from the pesticide residue in food and all other non-occupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure: Bacillus mycoides are commonly found in agricultural settings (Ref. 4) (i.e. water, soils and plant roots) and are present on fresh produce of all kinds with no known adverse effects. Bacillus mycoides isolate J is not known to produce mammalian toxins of foodborne concern and no foodborne disease outbreaks associated with Bacillus mycoides or toxin production from Bacillus mycoides in food or feed have been reported. 
Drinking Water Exposure: According to the World Health Organization, Bacillus species are often detected in drinking water even after going through acceptable water treatment processes, largely because the spores are resistant to these disinfection processes (Ref. 5). Should this microbial pesticide be present, no adverse effects are expected from exposure to Bacillus mycoides through drinking water, based on the results of toxicity studies described in Unit IV.
Other Non-occupational Exposure: Because Bacillus mycoides isolate J products do not currently include residential settings, exposure to the bacterium from its pesticide use would be very unlikely and the bacterium is already naturally-occurring in soils and water. Repeated exposures to the Bacillus mycoides isolate J microorganism from pesticidal applications do not exceed EPA's level of concern, particularly in light of available data that demonstrate Bacillus mycoides isolate J produced with the current manufacturing process is not toxic or pathogenic. (See additional discussion in Section II of this document).   Exposures other than those to workers, mitigated by label language for use patterns and Personal Protective Equipment (PPE), are not expected.  

1. Dermal exposure. Bacillus mycoides isolate J was shown to be non-toxic and not irritating to the skin (primary dermal irritation). (See additional discussion in Section II of this document).

 2. Inhalation exposure. Bacillus mycoides isolate J that was prepared with the current manufacturing process was shown to not be toxic or pathogenic. (See additional discussion in Section II of this document).


IV.  Cumulative Effects from Substances with a Common Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the EPA consider "available information concerning the cumulative effects of [a particular pesticide's] . . . residues and other substances that have a common mechanism of toxicity."
EPA has not found Bacillus mycoides isolate J to share a common mechanism of toxicity with any other substances, and Bacillus mycoides isolate J does not appear to produce toxic metabolites from normal exposures, like those produced by other substances. Thus, for the purposes of this tolerance action, the EPA has concluded that Bacillus mycoides isolate J does not have a common mechanism of toxicity with other substances. Following from this, therefore, the EPA concludes that there are no cumulative effects associated with Bacillus mycoides isolate J that need to be considered. 

For information regarding the EPA's efforts to determine chemicals that have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the EPA's website at http://www.epa.gov/pesticides/cumulative.

V.  Determination of Safety for the U.S. Population, Infants and Children

A.  U.S. Population, Infants and Children
No irritation or pathogenic endpoints were noted in mammalian testing of Bacillus mycoides isolate J and differential effects to infants and children are not expected. No relevant inhalation exposures to Bacillus mycoides isolate J resulting in pulmonary toxicity is expected from use on exempt treated foods. Bacillus mycoides is already very commonly present on a wide range of foods, including those consumed raw or with minimal processing.

VI.  Conclusions
The EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to residues of Bacillus mycoides isolate J. Therefore, an exemption from the requirement of a tolerance is established for residues of Bacillus mycoides isolate J in or on all agricultural commodities when used in accordance with label directions and good agricultural practices. 

In addition, the Agency is removing the existing paragraph contained in section 180.1269 because that tolerance for Bacillus mycoides isolate J has expired. 



VII.  References
 U.S. EPA. 2016, Memorandum from J. Gagliardi and John Kough, to S. Cerrelli, Re: Bacillus mycoides isolate J, Review of product characterization and a food tolerance exemption petition. Dated April 7, 2016.

 PMRA 2016, Jonas Ptasinski and Brian Belliveau, Data Evaluation Record, Re: Acute Pulmonary Infectivity and Toxicity.  Dated April 12, 2016.

 U.S. EPA. 2015, Joel Gagliardi (U.S.EPA) and Jonas Ptasinski [Canadian Pest Management Regulatory Agency (PMRA)] Review of Submitted Studies to Support FIFRA section 3 registration of a TGAI and EP and a new tolerance exemption, Dated July 14, 2015.

 Ludwig, W., Schleiffer, Whitman, W.H. (2009) Class I: Bacilli, class. nov., in "Bergey's Manual of Systematic Bacteriology", 2nd Edition, Volume 3, The Firmicutes, P. DeVos, G.M. Garrity, D. Jones, N.R.Krieg, W. Ludwig, K.H. Schleiffer, & W.B. Whitman (eds.) Springer Publishing.
4. 	World Health Organization, Guidelines for Drinking-Water Quality. (2011) Fourth Edition. http://www.who.int/water_sanitation_health/publications/2011/dwq_guidelines/en/


