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EPA REGISTRATION DIVISION COMPANY NOTICE OF FILING FOR 

PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER 

EPA Registration Division contact: [insert name and telephone number with area 

code] 

INSTRUCTIONS: Please utilize this outline in preparing the pesticide petition. In 

cases where the outline element does not apply, please insert “NA-Remove” and 

maintain the outline. Please do not change the margins, font, or format in your 

pesticide petition. Simply replace the instructions that appear in green, i.e., “[insert 

company name],” with the information specific to your action.

TEMPLATE: 

Koppert Biological Systems, Inc. 28465 Beverly Road Romulus, Michigan 48174

PP0E7727

EPA has received a pesticide petition PP0E7727 from Interregional Research 

Project Number 4 on behalf of Koppert Biological Systems, Inc. 28465 Beverly Road 

Romulus, Michigan 48174 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.

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

 lactoperoxidase (CAS No. 9003-99-0) under 40CFR 180.920 in or on all raw 

agricultural commodities when used pre-harvest as an inert ingredient in formulations of 

the active ingredient potassium iodide and potassium thiocyanate. EPA has determined 

that the petition contains data or information regarding the elements set forth in section 

408 (d)(2) of FDDCA; however, EPA has not fully evaluated the sufficiency of the 

submitted data at this time or whether the data supports granting of the petition. 

Additional data may be needed before EPA rules on the petition. 

A. Residue Chemistry

 1. Plant metabolism.

 2. Analytical method. Since it is proposed that lactoperoxidase be exempt from 

the requirement for a tolerance, no analytical method is necessary.

 3. Magnitude of residues. Since it is proposed that lactoperoxidase be exempt 

from the requirement for a tolerance, no analytical method is necessary.
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B. Toxicological Profile

 1. Acute toxicity. Lactoperoxidase is in the mammary, salivary and lachrymal glands of 

mammals and their respective secretions, e.g. milk, saliva and tears. Human salivary peroxidase 

has the same catalytic properties as bovine lactoperoxidase. Moreover the two enzymes have 

common antigenic determinants. Milk basic protein (MBP) is a well-characterized basic protein 

fraction derived from milk. MBP is approximately 97% total protein. Most of this protein is 

lactoferrin and lactoperoxidase, which comprise approximately 54% and 41% of the total MBP 

protein, respectively. No mortality was observed during the 14-day study period. There were no 

abnormalities in the general appearance in any animal, and no adverse changes in body weight 

occurred in any animal group during the study. Additionally, no organ pathology was observed at 

necropsy on Day 14. Accordingly, the median lethal dose (LD50) of MBP for the Crj:CD (SD) 

IGS rats in this test system was determined to be greater than 2000 mg/kg body weight.

 2. Genotoxicty. MBP was tested for the potential to induce point mutations in a reverse 

mutation assay using S. typhimurium strains TA98 and TA100. In both activated and nonactivated MBP-treated plates, the average number of revertant colonies was less than two-times 

the number of revertant colonies observed in the negative control plates, at incubation 

concentrations up to 5000 μg/plate. Results from the positive controls indicated appropriate 

sensitivity of the test system. There was no MBP concentration-dependent increase in revertants, 

and MBP-related inhibition of growth was not observed. Based on these results, under these 

testing conditions MBP was not mutagenic. (Kruger, 2007)

 3. Reproductive and developmental toxicity. There were no MBP-related adverse 

clinical effects observed over the course of the teratology study. There were no differences 

between treatment and control animals in body weight, body weight gain, food consumption, 

numbers of corpora lutea, numbers of implantation sites, numbers of live and dead fetuses, 

numbers of resorbed embryos, viability indices of fetuses, sex ratio, placental weight, and body 

weight of fetuses. In live fetuses, there were no significant MBP-related external, visceral, or 

skeletal anomalies or variations. Based on this study, under these test conditions MBP had no 

adverse effects on reproduction or development in Crj:CD (SD) 

IGS rats at 2000 mg/kg body weight/day.

 4. Subchronic toxicity. A subchronic 13 week rat oral toxicity study with MBP did not 

show any test article-related effects on clinical observations, body weight, feed efficiency, 

ophthalmic effects, hematology, necropsy and histopathology. 

Urinalysis results showed a dose-dependent increase in group mean sodium and chloride 

excretion in all animals. The values for sodium excretion, however, were well within the range of 

historical controls and the values for sodium excretion for the male and female control 

animals in this study were very low compared with background. Similarly, values for chloride 

excretion were well within the range of historical controls and the values for chloride excretion 

for the male and female control animals in this study were very low compared with background. 

Accordingly, the statistically significant differences in sodium and chloride excretion values in 

the treated animals were considered to be attributed to unusually low control values and not 

considered to be a biologically adverse finding. 

Compared with the control group, there were no statistically significant differences in blood 

chemistry values in male and female animals in the 200 mg/kg/day dose group or female animals 

in the 2000 mg/kg/day dose group. In male animals in the 2000 mg/kg/day dose group, there 

was a statistically significant differences in potassium concentration, compared with control 
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animals, however, this effect was not considered biologically significant because it only occurred 

in males, there was no change in urine potassium excretion values, and there were no 

abnormalities observed in the kidneys or adrenals in these animals. 

The no-observed adverse effect level (NOAEL) of MBP in rats was 2000 mg/kg body weight/day. 

 5. Chronic toxicity. Since lactoperoxidase is naturally produced in humans 

and animals, man has been chronically exposed without any known adverse effects.

 6. Animal metabolism. Since lactoperoxidase is naturally produced in humans 

and animals, any potential metabolites are naturally occurring.

 7. Metabolite toxicology. Since lactoperoxidase is naturally produced in 

humans and animals, any potential metabolites are naturally occurring. There are 

no known toxicological effects.

 8. Endocrine disruption. Not known to be an endocrine disruptor.

C. Aggregate Exposure

1. Dietary exposure. Lactoperoxidase is one of the most prominent enzymes naturally 

present in cow’s milk (containing between 24 and 30 mg/l). Only limited 

quantitative information was found on the presence of heme peroxidases in other 

human food sources. Obviously, food prepared from household mammals (cattle, 

sheep, goats, horses etc.) will contain heme peroxidases which are structurally related 

to the ones present in humans. Moreover, heme peroxidases are ubiquitously present 

in food plants, e.g., peanut peroxidases, horse-radish peroxidase, ascorbate 

peroxidases and yeast cytochrome C peroxidase (Bosshard et al., 1991; Dalton, 1991; 

Dunford, 1991; Huystee, 1991). The relevance of the plant heme peroxidases in the 

present context is limited due to structural differences with the bovine 

lactoperoxidase. It can be concluded that humans are exposed to a wide variety of 

heme peroxidases through their food. With the exception of milk, it is difficult to 

make reliable quantitative estimates of this exposure.

 i. Food. Lactoperoxidase is one of the major proteins present in whey and bovine milk 

basic protein (MBP). Various groups of whey proteins (including lactoperoxidase) and MBP are 

generally recognized as safe (GRAS) by the US FDA for the use in foods. Lactoperoxidase is 

recommended by the FAO/WHO as a safe method of preventing milk losses due to microbial 

spoilage. In New Zealand and Australia, LP is allowed as a processing aid in meats. 

 ii. Drinking water. No drinking water exposure is anticipated.

 2. Non-dietary exposure. Potential non-occupational exposure is not expected 

to pose any adverse effects to exposed populations via aggregate and cumulative 

exposures.

D. Cumulative Effects

For the purposes of this petition it is assumed that lactoperoxidase does not have a 
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common mechanism of toxicity with other substances.

E. Safety Determination

 1. U.S. population. Lactoperoxidase is in the mammary, salivary and lachrymal glands 

of mammals and their respective secretions, e.g. milk, saliva and tears. Human salivary 

peroxidase has the same catalytic properties as bovine lactoperoxidase. 

 2. Infants and children. Bovine LP behaves differently from other protective proteins 

(immunoglobulins, lysozyme, lactoferrin, and so on). Its concentration is low in the bovine 

colostrum and increases rapidly to reach a peak at 4-5 days postpartum. It declines rapidly 

afterwards to reach a constant, rather high, plateau during lactation. The human infant 

already produces salivary lactoperoxidase during the first few days after birth

F. International Tolerances

Lactoperoxidase is recommended by the FAO/WHO as a safe method of preventing milk 

losses due to microbial spoilage. In New Zealand and Australia, LP is allowed as a processing 

aid in meats.