Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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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 18th Street, NW, Suite 1000 

 Washington, DC 20036 

 
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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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 subtsugae is 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 

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SECTION B Amount, Frequency, and 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 select crops (corn, legumes, and cotton) on 

production agricultural crops, turf, nurseries, and commercial landscapes to control 

target insects (armyworms, cutworms, loopers, webworms, diamondback moth, aphids, 

billbugs, leafhoppers, mites, maggots, symphylans, thrips, whiteflies, beetles, 

wireworms, and root, knot, lesion, ring, sting, 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, cotton, 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 fields, 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 ranging from 2 to 8 ounces per gallon of water 

depending on pest pressure. 
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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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 toxicity/pathogenicity studies both in the rat, 

acute oral, dermal, and inhalation toxicity studies in the rat, and eye and dermal irritation 

studies in the rabbit. The oral, dermal, and inhalation routes of exposure 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-Insecticide 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 

LD50 > 5,000 mg/kg 

Category IV 

(December 10, 2009) 

Acute dermal toxicity 

MRID No. 49036906 

Rat Undiluted dose of 

5050 mg/kg 

LD50 > 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 

LC50 > 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 4.0x106 CFU/rat Non-toxic, non-pathogenic 

(August 4, 2010) 

Acute intravenous 

Toxicity/pathogenicity 

MRID No. 49036904 

Rat 1.8x107 CFU/rat Non-toxic, non-pathogenic 

(September 21, 2010) 
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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A waiver has been requested for acute oral toxicity/pathogenicity on the basis that 

existing studies (acute injection and pulmonary toxicity/pathogenicity and acute oral 

toxicity) demonstrate the non-toxic and non-pathogenic nature of this product across all 

acute exposure pathways. 

 

The results of toxicity testing on the organism 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 there are no reports of ecological or human 

health hazards caused Chromobacterium subtsugae. It does not produce recognized 

toxins, enzymes, or virulence factors normally associated with mammalian invasiveness 

or toxicity. 

 

As has been demonstrated, dietary exposure to Chromobacterium subtsugae strain 

SB3872 meets the FQPA 1996 safety standard. Studies 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 with mammalian invasiveness or toxicity. 

The absence of acute toxicity or pathogenicity in laboratory animals 

demonstrates the benign nature of this strain. 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 water 
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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(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. Additionally the bacteria would not tolerate 

the conditions water is subjected to in a drinking-water treatment facility 

(including: chlorination, pH adjustments, 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 with mammalian invasiveness or toxicity. 

The absence of acute toxicity or pathogenicity in laboratory animals 

demonstrates the benign nature of this strain. 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 demonstrates 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 strain. 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 and 
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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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 absence 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 ecological 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 expected that, when used as proposed, 

Chromobacterium subtsugae strain SB3872 would result in residues that are of 

toxicological concern. There is a reasonable 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 

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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 residues 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 

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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 

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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 tolerance exemption in 40 CFR §180.1305 for Chromobacterium 

subtsugae strain PRAA4-1T: 

“An exemption from the requirement 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 accordance with good agricultural practices.”
Chromobacterium subtsugae strain SB3872 – Petition to Establish a Tolerance Exemption 

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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 tolerance exemption for the similar Chromobacterium 

subtsugae strain PRAA4-1T, codified under 40 CFR §180.1305, which states: 

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

Chromobacterium subtsugae strain PRAA4-1T 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 Residue 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 absence of acute toxicity or pathogenicity in 

laboratory animals demonstrates the benign nature of this strain, and with the 

established tolerance exemption for a similar Chromobacterium active ingredient, 

serves to demonstrate reasonable grounds in support of the petition for exemption from 

the requirement of a tolerance.