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EPA REGISTRATION DIVISION CO MPANY 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 p reparing the pesticide petition. In

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

maintain the outline. Please do not chan ge 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 pot assium 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 la ctoperoxidase 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.2



### 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 l actoperoxidase. Moreover the two enzymes have common antigenic determinants. Milk basic prot ein (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 an imal, 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 le thal 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 μ g/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 th e 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 implanta tion sites, numbers of live and dead fetuses, numbers of resorbed embryos, vi ability indices of fetuses, sex ra tio, 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 th is 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 clini cal 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 ex cretion, 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 w ith 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 differen ces in sodium and chloride excretion values in

the treated animals were considered to be a ttributed 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 control3

animals, however, this effect was not consider ed biologically significant because it only occurred in males, there was no change in urine po tassium 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 na turally produced in humans and animals, man has been chronically ex posed without any known adverse effects.

6. Animal metabolism. Since lactoperoxidase is naturally produced in humans and animals, any potential meta bolites 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 pe roxidases which are structurally related to the ones present in humans. Moreover, heme peroxidases are ubiquitously present in food plants, e.g., peanut peroxi dases, horse-radish peroxidase, ascorbate peroxidases and yeast cytochrome C peroxidase (Bosshard et al., 1991; Dalton, 1991; Dunford, 1991; Huystee, 1991). The releva nce of the plant heme peroxidases in the present context is limited due to struct ural 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 est imates of this exposure.



### i. Food. Lactoperoxidase is one of the major prot eins 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 F DA 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 a ssumed that lactoperoxidase does not have a4



### 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 Zeal and and Australia, LP is allowed as a processing

aid in meats.