Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 1 of 11 

## Petition for an exemption from the requirement of a tolerance for residues of products containing the active ingredient

“ Chromobacterium subtsugae strain SB3872” in and on all

food commodities

Submitted for: Novozymes BioAg, Inc. (EPA Company No. 73314) 13100 W. Lisbon Road, Suite 600 Brookfield, Wisconsin 53005

Submitted by: Technology Sciences Group Inc.

1150 18 th Street, NW, Suite 1000 Washington, DC 20036Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 2 of 11 

## SECTION A Name, Chemical identity, and Composition of the

Pesticide Chemical

Product Name: ChromoTM Bio-Insecticide TGAI

ChromoTM Bio-Insecticide EP

Active Ingredient: Chromobacterium subtsugae strain SB3872

Chromobacterium subtsugaeis a gram-negative, violetpigmented bacterium, which was described in a recent publication as being isolated from forest soil rich in hemlock

leaves from the Catoctin Mountain region of central Maryland (Martin et al. 2007). Chromobacterium subtsugae is a naturally occurring soil bacterium that is found in soils worldwide. Strains of this organism are generally not regarded as human or animal pathogens.

The same formulation as the TGAI is to be registered as an end-use product to control a variety of target insect pests (i.e. caterpillars, aphids, whiteflies, coleopteran, and plant sucking mites) on growing agricultural crops, greenhouses, turf, commercial landscapes, and residential homes and gardens.

Mode of Action:

The mode of action of this strain has been demonstrated in several studies by Phyllis Martin et.

al. 2007. Chromobacterium subtsugae was found to be orally toxic to Colorado Potato Beetle ( Leptinotarsa

decemlineata ) larvae, Small Hive Beetle ( Aethina

tumida ) larvae, and Southern Corn Rootworm ( Diabrotica undecimpunctata ) larvae. In addition, the

microbe was also found to be toxic to adult D. undecimpuntata and adult Southern Green Stinkbugs ( Nezara viridula ) after consuming Chromobacterium subtsugae in water or bail diet (Martin et al. 2007).Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 3 of 11 

## SECTION B Amount, Frequency, a nd Timing of Application of

the Pesticide Chemical

The intended use of Chromobacterium subtsugae strain SB3872 in ChromoTM BioInsecticide is by foliar spray or soil drench application (via ground or aerial), by chemigation, and/or by seed treatment for se lect crops (corn, legumes, and cotton) on production agricultural crops, turf, nurseries, and commercial landscapes to control target insects (armyworms, cutworms, l oopers, webworms, diamondback moth, aphids, billbugs, leafhoppers, mites, maggots, sym phylans, thrips, whiteflies, beetles, wireworms, and root, knot, lesion, ring, st ing, and stunt nematodes, among others). ChromoTM Bio-Insecticide can be applied at 3-14 day intervals at rates of 1⁄4 to 2 gallons per 100 gallons of water or 1⁄4 to 2 gallons per acre to alfalfa, forage crops, artichoke, asparagus, banana, brassica (cole) leafy vegetables, bulb vegetables, bushberries, caneberries, cereal grains, citrus, corn, cott on, cranberry, cucurbits, fig, ornamental plants, fruiting vegetables, grape, herbs and spices, hops and dried cones, leafy vegetables, root and tuber vegetables, legume vegetables, oilseed crops, peanut, peppermint, pineapple, pome fruit, pomegranate, shade and ornamental trees, stone fruit, strawberry, sugarcane, tobacco, tree nuts, pistachios, tropical/subtropical fruit, and turf up to and including the day of harvest. Post-harvest applications can be made at a rate of 1-2 gallons per 100 gallons of water.

Also, ChromoTM Bio-Insecticide can be applied to poultry houses to control flies and

litter beetles and to exterior structures to control boxelder beetle and brown marmorated stink bug at rates of 1⁄4 to 2 gallons per 12 gallons of water.

ChromoTM Bio-Insecticide can be applied to golf course tees, greens, and fairways, commercial and residential lawns, athletic fi elds, parks, cemeteries, turf landscapes, herbaceous and woody ornamentals, trees and plants against armyworms, cutworms, webworms, chinch bugs, leafhoppers, white grubs , beetles, chafers, loopers, leafrollers, moths, caterpillars, nematodes, billbugs and larvae at rates of 1⁄4 to 2 gallons per acre at the same application interval (3-14 days) as crop use.

For residential/home and garden use, ChromoTM Bio-Insecticide can be applied as a foliar spray to control soil-borne (maggots, wireworms, symphylans, cutworms, white grubs, nematodes) and turf-borne (webworms, cutworms, chinch bugs, leafhoppers) pests on vegetables, fruits, nuts, ornamental plants, trees, shrubs, flowers, foliage, and tropical plants every 3-10 days at rates rangi ng from 2 to 8 ounces per gallon of water depending on pest pressure.Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 4 of 11 

## SECTION C Reports on Investigations Made with Respect to the Safety of the Pesticide Chemical

Toxicity of Chromobacterium subtsugae strain SB3872

Studies to evaluate the safety to mammals were conducted on the technical grade active ingredient Chromobacterium subtsugae strain SB3872 and are summarized herein (see Table 1). The studies performed include acute pulmonary toxicity/pathogenicity and acute injection toxi city/pathogenicity studies both in the rat, acute oral, dermal, and inhalation toxicity st udies in the rat, and eye and dermal irritation studies in the rabbit. The oral, dermal, and i nhalation routes of ex posure are the routes

relevant for human health risk assessment. The results of these studies consistently

showed that Chromobacterium subtsugae strain SB3872 was non-toxic and also lacked irritation potential. Additionally, there was no evidence of pathogenically observed; the product was no infective or pathogenic to test animals when administered via the intravenous or pulmonary routes. No reports of hypersensitivity have been recorded from personnel working with this organism.

Table 1. Summary of CHROMO Bio-Insect icide TGAI Mammalian Toxicity Data Test Species Dose Tested Results (Date Completed) Acute oral toxicity

MRID No. 49036905 Rat Gavage dose of 5,000 mg/kg LD 50 > 5,000 mg/kg Category IV (December 10, 2009) Acute dermal toxicity

MRID No. 49036906 Rat Undiluted dose of 5050 mg/kg LD 50 > 5,050 mg/kg Category IV (December 2, 2009)

Acute inhalation toxicity

MRID No. 49036907 Rat >2.21 mg/L for 4

hours Non-toxic

LC 50 > 2.21 mg/L (December 8, 2009) Primary eye irritation

MRID No. 49036908 Rabbit 0.1 mL undiluted product for 24

hours Practically non-irritating Category IV (November 20, 2009)

Dermal irritation

MRID No. 49036909 Rabbit 0.5 mL undiluted product for 4 hours Non-irritating Category IV (December 3, 2009) Acute pulmonary toxicity/pathogenicity

MRID No. 49036903 Rat 4.0x10 6 CFU/rat Non-toxic, non-pathogenic (August 4, 2010)

Acute intravenous Toxicity/pathogenicity

MRID No. 49036904 Rat 1.8x10 7 CFU/rat Non-toxic, non-pathogenic (September 21, 2010)Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 5 of 11 A waiver has been requested for acute oral toxicity/pathogenicity on the basis that existing studies (acute injection and pulm onary toxicity/pathogenicity and acute oral toxicity) demonstrate the non-toxic and non-pat hogenic nature of this product across all acute exposure pathways.

The results of toxicity testing on the organi sm show there is no risk to human health from the active ingredient. Chromobacterium subtsugae strain SB3872 is not toxic, pathogenic, infective, or irritating to mammals.

Literature searches have demonstrated that t here are no reports of ecological or human

health hazards caused Chromobacterium subtsugae . It does not produce recognized toxins, enzymes, or virulence factors no rmally associated with mammalian invasiveness or toxicity.

As has been demonstrated, dietary exposure to Chromobacterium subtsugae strain

SB3872 meets the FQPA 1996 safety standard. St udies submitted with this petition showed no evidence of toxicity, pathogenicity, or infectivity when this strain was tested in laboratory animals.

Aggregate Exposure

1) Dietary Exposure:

Dietary exposure from use of Chromobacterium subtsugae strain SB3872, as proposed, is minimal. The intended use of Chromobacterium subtsugae strain SB3872 is to growing agricultural crops, commercial turf and landscapes, and residential lawns and gardens for the purposes of insect control.

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

environment from Chromobacterium subtsugae strain SB3872. There are no reports of ecological or human health hazards caused by the microorganism Chromobacterium subtsugae . It does not produce recognized toxins, enzymes, or virulence factors normally associated wit h mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this st rain. Dietary exposures would not be expected to pose any quantifiable risk, due to a lack of residues of toxicological

concern.

2) Drinking Water Exposure:

Similarly, exposure to humans from residues of Chromobacterium subtsugae strain SB3872 in consumed drinking water would be unlikely. Chromobacterium subtsugae is not known to grow or thrive in aquatic environments. Potential exposure to surface water would be negligible and exposure to drinking waterChromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 6 of 11 (well or ground water) would be impossible to measure. The intended use of Chromobacterium subtsugae strain SB3872 is to growing agricultural crops, commercial turf and landscapes, and residential lawns and gardens for the purposes of insect control. The risk of the microorganism passing through the soil to ground water is minimal to unlikely. A dditionally the bacteria would not tolerate the conditions water is subjected to in a drinking-water treatment facility (including: chlorination, pH adjustment s, high temperatures and/ or anaerobic conditions).

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

environment from Chromobacterium subtsugae strain SB3872. There are no reports of ecological or human health hazards caused by the microorganism Chromobacterium subtsugae . It does not produce recognized toxins, enzymes, or virulence factors normally associated wit h mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this st rain. Drinking water exposures would not be expected to pose any quantifiable risk, due to a lack of residues of toxicological concern.

3) Non-Dietary Exposure:

The potential for non-dietary exposure to the general population, including infants and children, is unlikely. The intended use of Chromobacterium subtsugae strain SB3872 is to agricultural crops, commercial turf and landscapes, and residential lawns and gardens for the purposes of insect control. The non-toxic and nonpathogenic nature of this product demonstrat es that other incidental non-dietary exposures are not of concern.

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

environment from Chromobacterium subtsugae strain SB3872. There are no reports of ecological or human health hazards caused by the microorganism Chromobacterium subtsugae. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this st rain. Non-dietary exposures would not be expected to pose any quantifiable risk due to a lack of residues of toxicological concern.

Cumulative Effects

It is not expected that, when used as proposed, Chromobacterium subtsugae strain

SB3872 would result in residues that are of toxicological concern. The intended use of Chromobacterium subtsugae strain SB3872 is to agricultural crops, commercial turf andChromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 7 of 11 landscapes, and residential lawns and gardens for the purposes of insect control. The results of toxicity testing indicate there is no risk to human health or the environment

from Chromobacterium subtsugae strain SB3872. There are no reports of ecological or human health hazards caused by Chromobacterium subtsugae strain SB3872. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absenc e of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain. Any exposures would not be expected to pose any quantifiable risk due to a lack of residues of toxicological

concern.

Safety Determination

1) General US Population:

Acute toxicity studies have demonstrated that Chromobacterium subtsugae strain SB3872 is not toxic, pathogenic, infective, or irritating to mammals. The intended

use of Chromobacterium subtsugae strain SB3872 is to agricultural crops, commercial turf and landscapes, and residential lawns and gardens for the purposes of insect control. The results of toxicity testing indicate there is no risk

to human health or the environment from Chromobacterium subtsugae strain SB3872. There are no reports of ecologi cal or human hazards caused by the microorganism Chromobacterium subtsugae . It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this strain. There is a reasonable certainty of no harm to the general US population from exposure to this active ingredient.

2) Infants and Children:

As mentioned above, it is not expec ted that, when used as proposed, Chromobacterium subtsugae strain SB3872 would result in residues that are of toxicological concern. There is a reas onable certainty of no harm for infants and children from exposure to Chromobacterium subtsugae strain SB3872 from the proposed uses.

Information on Endocrine Effects

To date there is no evidence to suggest that Chromobacterium subtsugae strain SB3872 functions in a manner similar to any known hormone, or that it acts as an endocrine disrupter.Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 8 of 11 

## Section D Tests on the Amount of Residue Remaining

Not applicable, Novozymes BioAg, Inc. is petitioning for an exemption from the requirement

of a tolerance for residues of Chromobacterium subtsugae strain SB3872.

Analytical Method: An analytical method for residue s is not applicable. It is expected that, when used as directed, Chromobacterium subtsugae strain SB3872 will not result in residues that are of toxicological concern.Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 9 of 11 

## SECTION E - Practical Methods for Removing Residues

Not applicable, Novozymes BioAg, Inc. is petitioning for an exemption from the requirement

of a tolerance for residues of Chromobacterium subtsugae strain SB3872.Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 10 of 11 

## SECTION F – Proposed Tolerance Exemption

Novozymes BioAg, Inc. is proposing that the following Exemption from the need of a tolerance be established for the pesticide, Chromobacterium subtsugae strain SB3872,

similar to the current toleranc e exemption in 40 CFR §180.1305 for Chromobacterium subtsugae strain PRAA4-1 T :

“An exemption from the requi rement of a tolerance is established for residues of Chromobacterium subtsugae strain SB3872 in or on all food commodities when applied as

an insecticide or miticide and used in a ccordance with good agricultural practices.”Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption Page 11 of 11 

## SECTION G Reasonable Grounds in Support of an Exemption

from the Need of a Tolerance

EXISTING TOLERANCES

US EPA Tolerance -

There is an established and existing to lerance exemption for the similar Chromobacterium subtsugae strain PRAA4-1 T , codified under 40 CFR §180.1305, which states:

“An exemption from the requi rement of a tolerance is established for residues of Chromobacterium subtsugae strain PRAA4-1 T in or on all food commodities when applied as an insecticide or miticide and used in accordance with good agricultural practices.”

International -

A Codex Alimentarium Commission Maximum Re sidue Level (MRL) is neither established nor required for Chromobacterium subtsugae strain SB3872.

It is not expected that, when used as proposed, Chromobacterium subtsugae strain

SB3872 would result in residues that are of toxicological concern. The intended use of Chromobacterium subtsugae strain SB3872 is to agricultural crops, commercial turf and landscapes, and residential lawns and gardens for the purposes of insect control. The results of toxicity testing indicate there is no risk to human health or the environment

from Chromobacterium subtsugae strain SB3872. There are no reports of ecological or human health hazards caused by Chromobacterium subtsugae strain SB3872. It does not produce recognized toxins, enzymes, or virulence factors normally associated with mammalian invasiveness or toxicity. The absenc e of acute toxicity or pathogenicity in laboratory animals demonstrates the beni gn nature of this strain, and with the established tolerance exemption for a similar Chromobacterium active ingredient, serves to demonstrate reasonable grounds in s upport of the petition for exemption from the requirement of a tolerance.