2 ISKBC Comment: Section 4.a. Ecological Risk Assessment, first paragraph, "EFED evaluated cyazofamid's potential risks to non

- target and endangered species for aquatic and terrestrial organisms (including plants) (DP Barcodes D3 1 9107+ and D356880+). In addition, in the 2012

assessment EFED evaluated chronic risk to b irds and risk to terrestrial plants, based upon newly reviewed data (DP Barcode D396387).

We have not received the DERs for any of the studies that were apparently reviewed and referenced here. Please provide the 2012 EFED assessment (DP Barcode D396387 ) and DERs

for the submitted studies.

EFED Response:

EFED has supplied the requested 2012 EFED assessment (D396387) and DERs listed in the

table below.

DP Barcode MRID Study Type Executive Summary from DER

D366145 47716308 850.2300

Colinus virginianus The one

- generation reproductive toxicity of IKF

- 916 Technical (cyazofamid) to 24 pairs per level (48 for the control) of 27

- week old Norther bobwhite quail (Colinus virginianus) was assessed over ca. 22 weeks. IKD

- 916 Technical was administere d to the birds in the diet at nominal concentrations of 0 (control), 28, 84, 167, or 1000 mg/kg diet (not adjusted for purity). Mean measured concentrations were <7.14 (<LOD, control), 28.3, 86.1, 170, and 1020 mg ai/kg diet, respectively.

There were no treatment

- related effects observed on any adult parameter. Egg production was the only reproductive parameter evaluated and this endpoint was reduced in a dose

- dependent manner 17 to 25%, relative to control; the reviewer’s analysis detected a

statistical ly significant difference (p=0.035) at only the highest treatment level. Given this reduction, the reviewer determined the NOAEC to be 170 mg ai/kg diet.

This study is scientifically sound and does satisfy the guideline requirement for a Northern bobwhite ( Colinus virginianus) reproductive study.

D366145 47716309 850.4100 Seedling

emergence The effect of IKF

- 916 400SC (ai cyazofamid) on the seedling emergence of monocot (corn, Zea mays; oat, Avena sativa ; onion, Allium cepa ; and ryegrass) and dico t (cucumber, Cucumis sativa; lettuce, Lactuna sativa ; carrot, Daucus carota; soybean, Glycine max; radish, Raphanus sativus ; cabbage, Brassica oleracea; and tomato, Lycopersicon esculentum ) crops were studied at nominal concentrations of 0 (negative contro l) and 1.56 lbs ai/A (equivalent to 4.65 lbs formulation/A).

The growth medium used in the seedling emergence test was a mixture of collected silt loam and masonry sand that was classified as3 a loam soil (pH 7.3,% organic matter 1.5%). After 14 days, the number of plants emerged was recorded and taken for measuring dry weight and height.

Negative control emergence ranged from 80 to 100% for all species tested, and treatment group emergence ranged from 86% to 100%. There was promotion of growth in seedling height for carrot, oat and radish, while inhibitions for all other species ranged fr om 1 to 6%.

There was promotion of growth in dry weight for cucumber, corn, oat, and radish, while cabbage and ryegrass did not experience inhibitions in dry weight. Lettuce, onion, soybean and tomato had inhibitions ranging from 2% to 11% and carrot h ad an inhibition in dry weight of

19%.

No phytoxicity or abnormality was reported for any species.

The most sensitive monocot and dicot species could not be determined due to the lack of a statistically significant effect on any of the plants. The over all NOAEC and EC 25 values for all species tested were 1.56 and >1.56 ai/A. respectively, which is equivalent to 4.65 and >4.65 lbs formulation/A, respectively.

This toxicity study is classified as acceptable and satisfied the guideline requirement for a T ier I terrestrial plant toxicity study.

D366145 47716310 850.4150 Vegetative Vigor The effect of IKD

- 916 400SC (ai cyazofamid) on the vegetative vigor of monocot (corn, Zea mays; oat, Avena sativa ; onion, Allium cepa ; and ryegrass) and dicot (cucumber, Cu cumis sativa; lettuce, Lactuna

sativa ; carrot, Daucus carota; soybean, Glycine max; radish, Raphanus sativus ; cabbage, Brassica oleracea; and tomato, Lycopersicon esculentum ) crops were studies at nominal concentrations of 0 (negative control) and 1.56 lbs ai/A. Corn, cucumber, lettuce, oat, onion and soybean were actually treated with 1.56 lbs cyazofamid/A, while cabbage, carrot, radish, ryegrass and tomato were treated with 1.57 lbs cyazofamid/A (equivalent to 4.65 and 4.68 lbs formulation/A, respectivel y)

The growth medium used in the seedling emergence test was a mixture of collected silt loan and masonry sand that was classified as a loam soil (pH 7.3, % organic matter 1.5%). After 14 days all plants were recorded and taken for measuring dry weight and height.

Height and dry weight per plant were unaffected by the test material.

No phytotoxicity or abnormality was reported for any species.4 The most sensitive monocot and dicot species could not be determined due to the lack of a statistically signi ficant effect on any of the plants. The overall NOAEC and EC 25 values for corn, cucumber, lettuce, oat, onion and soybean were 1.56 and >1.56 ai/A respectively, which is equivalent to 4.65 and >4.65 lbs formulation/A, respectively. The overall NOAEC and EC 25 values for cabbage, carrot, radish, ryegrass and tomato were 1.57 and >1.57 lbs ai/A respectively, which is equivalent to 4.68 and 4.65 lbs formulation/A.

This study is classified as acceptable and satisfied the guideline requirement for a Tier I Ter restrial Plant Vegetative Vigor toxicity study.

ISKBC Comment: Page 18, 6 Receptors c. Ecological Incidents : According to the Agency's records, as well as ISK Biosciences Corporation's records, there have been no ecological

incidents associated with cyazofamid. Thus this fact provides some degree of certainty that the models being used that suggest potential concerns with non

- target organisms can be considered very conservative.

EFED Response: EFED is aware that there have been no ecological incidents associated with cyazofamid and have thus reported accordingly in the Problem Formulation. At the conclusion of the above mentioned paragraph which details the databases used in incident compilatio n
, the Problem

Formulation states, “EIIS, AIMS, and Aggregates Incident Database were run on November 12, 2014 and did not return any incidents.” Incident occurrence is used in risk characterization to demonstrate a complete exposure pathway, however the absence of reported incidents does

not indicate that there is an absence of exposure. The absence of reported incidents does not change the potential concerns for non

- target organisms nor does it indicate whether or not

EFED models used to assess risk to n on

- target organisms are conservative.

ISKBC Comment: Page 19, 8. Analysis Plan, a. Stressors of Concern, The stressors of concern are listed as cyazofamid and degradation products (CCIM, CCIM

- AM and CTCA). "Submitted

studies on mammals indicate that CC IM toxicity to mammals is more toxic than cyazofamid; this increased toxicity triggers concern on an acute basis. CCIM

- AM and CTCA have similar toxicity to the parent.

This conclusion was based on one acute oral study of CCIM. HED has also identified t hese degradation products in the "Cyazofamid Human Health Scoping Document in Support of Registration Review" as residues of concern in water. Only parent and CCIM were of concern in plant residues. In that same document, HED stated they did not have any concerns to the population from exposure to parent or its degradation products. They further stated in the review table on page 24 that the results of the special metabolism studies suggest that "IKF

- 916

is stable in stomach contents" of rats and "CCIM c ould be produced in rats following IKF

- 916 entering the blood compartment but not necessarily in the stomach compartment." Thus, the5 ecological concern for the formation of CCIM from exposure to consuming cyazofamid from plant tissues should be greatly re duced.

EFED Response:

All s oil metabolism studie s demonstrate the formation of CCIM from the parent in high concentrations; formation is expected to occur in plant tissue as well. Toxicity data demonstrates CCIM to have a higher toxicity than the parent. Additionally, CCIM is highly persistent in water (stable to hydrolysis) and moderate in soils, ~0.9 days to ~2.9 days. There is the potential for non

- target terrestrial organisms to come into contact with CCIM on the field, therefore the ecological concer n for CCIM remains.

The section quoted in the above comment from ISKBC, "CCIM could be produced in rats following IKF

- 916 entering the blood compartment but not necessar ily in the stomach compartment,” is referring to the likelihood of cyazofamid degrad ing to CCIM while in the stomach compartment of the organism. While this particular route of exposure is not likely, CCIM presents other route of exposure as well. T here is the potential for CCIM to be present in plant tissues consumed by mammals and avian species , thus exposure cannot be assumed to

be reduced.

ISKBC Comment: Page 25, Section 11. Preliminary Identification of Data Gaps, a.

E nvironmental Fate Table 4, (also mentioned on page 2

- Data Gaps, Environmental Chemistry Methods/Independent Laboratory Validations); OCSPP 850.6100: Analytical methods in soil, water, and sediment are required.

Methods for water and soil were submitted (MRID Nos. 45409021 & 45409037) and this review suggests that they are still in review, thus, we have not re ceived any DERs to which we can respond to comments that suggest the data is not acceptable. No new studies should be required until we have an opportunity to review and correct any deficiencies in these studies. Please provide the DERs for these studies . We have a study in sediment that was submitted to and accepted by Canada and we will submit that study to the Agency to address the method for sediment. Thus we request a waiver from conducting any further studies for this requirement until the Agency completes their review of the submitted studies.

EFED Response: As part of the screen EFED conducted of these ECM/ILV’s , a number of issues were observed, which were named in the Problem Formulation. It was further indicated that

the studies were in revi ew. EFED completed the screen of the studies and found the studies to

be unreviewable since the ECMs were submitted in the same document than the ILV s . A separate memorandum will be issued for these studies. In order to consider the studies, separate

EC Ms and ILVs should be submitted. Other issues noted in the stu dies should also be addressed

at the time the studies are resubmitted.

ISKBC Comment: Page 27, Section 11. Preliminary Identification of Data Gaps, b. Effects, Table

6

- Data Needs, fish toxicity, invertebrate life cycle, saltwater invertebrates (OCSPP 850.1075; 1300; 1350)6 Estuarine/marine fish Toxicity; The DER stated, "This study is scientifically sound, but it is classified as Supplemental, because terminal fish weights ranged lower than the initial fish weight that is recommended by USEPA. The 96

- hour LC 50 was >167 μ g a.i./L, the highest

level tested and above the water solubility level, which categorizes IKF

- 916 Technical as highly toxic to sheepshead minnow on an acute toxicity ba sis. The ECOSAR model estimates the

LC 50 in fish to be 2.551 mg/L, which is over 100 fold safer than the study value. Since this was an acute study, repeating the study will most likely not change the LC 50 value for this requirement due to solubility lim its and we request a waiver from conducting a replacement study for this requirement.

Invertebrate life cycle; Two studies were submitted for this requirement. In the first study, (MRID # 45409136) the author reported the NOEC as 0.039 mg a.i./L with the LOEC being >0.11 mg a.i./L. The Agency's DER stated, "The study is scientifically sound, but it does not

fulfill the USEPA requirements for a chronic freshwater invertebrate toxicity study (Subdivision E, §72

- 4b) because dry weights were not measured. A s a result, this study is

classified as INVALID." In the second study (MRID 46702708), the DER classified the study

as supplemental "based on the conclusion that the NOAEC for dry body weight was below the lowest rate tested (0.0087 mg ai/L)". A respons e to the DER was submitted on 3/26/09 (MRID 47716307) in which the l aboratory conducting the study addressed EPA's comments and provided justification that the NOAEC for dry body weight was 0.02 mg a

. i

. /L, which is similar

to the first study. Additionally , based on the well

- defined concentration response provided in this study, it is unlikely that a repeat test will define the NOAEC any more precisely than provided by this value. We have not seen a DER for this response and we note that it is not

mentioned in the document. Thus, we request a waiver from conducting any further studies for this requirement until the Agency completes their review of the submitted response.

EFED Response: The DER for MRID 45409131 on the acute toxicity of cyazofamid to Sheep shead minnow (Cyprinodon variegatus) classified the study as supplemental. The terminal fish weights for the study were 0.46 g, and ranged lower than the minimum initial fish weight that is recommended by USEPA (0.5 g). Individual fish weights were not p rovided, so the variation and fish weight and thus the severity of this deviation cannot be adequately determined. Because fish weight can affect fish response to pesticides, this deviation impacted the acceptability of the study. The study saw 15% morta lity in the highest treatment level, which is equivalent to the solubility limit. A study waiver may be considered for the study gap if the individual fish weight data for this study is submitted, reviewed, and the variation in the fish weight is determin ed to not be severe enough to be considered a study deficiency.

Two studies were available for the daphnid full life cycle guideline requirement: MRID 45409136 was classified as invalid based on lack of measurement of dry weights and MRID

46702708 was cl assified as supplemental because the NOAEC for dry weights was below the lowest rate tested (0.0087 mg ai/L). A follow up to MRID 46702708 was submitted under MRID 47716307 in March 2009; EFED responded to this follow up in August 2010 under

D366145. EFE D stated the following in the memorandum:7 “ This study was classified previously as SUPPLEMENTAL because a valid NOAEC could not be establish as well as a question regarding a drop in toxicant concentrations due to a problem with the toxicant delivery pump . The NOAEC was determined to be <0.0087 mg ai/L. In the response, it was suggested that pooled control data could be used for NOAEC determination as well as substituting an EC10 for a NOAEC per the updated OECD Algal Test Guideline #201. The response

s uggested that the EC10 was 0.019 mg ai/L with 95% confidence limits of 0.0085 to 0.41 mg ai/L. ”

Additionally, t he DER for this response has been provided. Daphnid full life cycle is still considered to be a data gap.

ISKBC Comment: Page 28, Section 11. Preliminary Identification of Data Gaps, b. Effects, Table

6

- Data Needs, Cyanobacteria (OCSPP 850.4550)

A Cyanobacteria study (MRID 46209402) was submitted to the Agency which was listed as

invalid. Three other algae studies were also listed as invalid. The NOAEC was determined to

be >0.25 mg a.i./L, the highest level tested. The ECOSAR model estimates the NOAEC in

green algae to be 0.186 mg/L, which is very similar the study value. Since we have not received DERs for any of th ese studies, replacement studies should not be required until we have received the DERs and have had a chance to provide additional information to correct any

deficiencies. Please provide the DERs for the submitted studies so that we can respond to the

re views. Thus we request a waiver from conducting any further studies for this requirement at

this time.

EFED Response:

E FED has provided the DERs for the following MRIDs, the executive summaries have been

summarized in the table below. Of the submitted s tudies, three were classified as supplemental, one was classified as invalid and one was classified as acceptable.

DP Barcode MRID Study Type Executive Summary from DER

D304897 46209402 850.45 00

Skeletonema

costatum In a 96

- hour acute toxicity study, culture of Skeletonema costatum were exposed to Cyazofamid under static conditions at nominal concentrations 0 (negative and solvent controls), 0.010, 0.026, 0.064, 0.16, 0.40, and 1.0 ppm ai. The time weighted mean

meas ured concentrations were <0.0021

- 0.0025 (<LOQ, negative and solvent controls), 0.0036, 0.0088, 0.020, 0.045 , 0.087 and 0.10 ppm ai. At the 0

- hour analytical determination, only the two lowest treatment concentrations produced recoveries >70% of nominal. Furthermore, the three highest nominal concentrations exceeded the limit of water solubility (0.108 mg/L) for Cyazofamid Technical. All concentrations of test material declined substantially over the course of the study, presumably due to the photo degrad ation of cyazofamid and to the presence of algae.8 The toxicity study is scientifically sound but it does not satisfy the US EPA Guideline Subdivision J, 123

- 2 because of the uncertainty associated with the actual test concentrations that algae were

expo sed to, and thus, the inability to derive accurate toxicity values. As a result, the study is classified as supplemental.

D283446 45409143 850.45 00

S elenastrum capricornutum In a 96

- hour acute toxicity study, cultures of Selenastrum capricornutum were ex posed to Cyazofamid under static conditions. Nominal concentrations were 5, 10, 22, 46 and 100 μg/L. The mean measured concentrations for the two highest treatment concentrations ranged below 70% of nominal over the 96

- hour study

and measurement of the l owest treatment concentration was inaccurate. Cell density, biomass, and growth rate were significantly affected by treatment with cyazofamid. Cell growth inhibition ranged from 18.6% in the nominal 5 μg/L treatment group to 72.1 % in the nominal 100 μg/L treatment group.

The study is not scientifically sound and does not satisfy the US EPA Guideline Subdivision J, 123

- 2 for an aquatic nonvascular plant study. The initial 24 hours of the test were conducted in darkness

because the test material was con sidered photosensitive, impurities (detected during analytical determination) were introduced during sample pretreatment and measurement of the 4.8 μg/L solution concentrations were inaccurate and, therefore, calculation by extrapolation. As a result, a r eliable toxicity value could not be estimated from this study and it is classified as invalid.

D304897 46258213 850.45 00

Selenastrum capricornutum In a 96

- hour acute toxicity study, cultures of Selenastrum capricornutum were exposed to Cyazofamid (IKF

- 916 400SC with organosilicone adjuvant) under stat ic conditions at nominal concentrations of 0 (negative control), 4.6, 40, 22, 46, 100, 220, 460 and 1000 ppb ai (corresponded to 12.8, 28, 61, 128, 280, 610, 1280, and 2800 ppb). Because the measured concentr ations declined

>70% of nominal over the course of the study, the measured 0

- hour concentrations were used to compute the toxicity values; the 0

- hour concentrations were 4.0, 10, 22, 43, 98, 221, 458 and 1000 ppb ai. During the test, continuous lighting w as provided from 24 to 96 hours, but no lighting was used in the first 24 hours. The 96

- hour cell density percent inhibitions were 11, 28, 25, 38, 59, 58, 66 and 68% in the treatment groups respectively. The biomass percent inhibitions were 9, 21, 24, 3 6, 61, 67, and 19% in the treatment groups respectively; The growth rate (24 to 96 hours) percent inhibitions were 3.8, 7.9, 7.1, 9.6, 16.9, 19.6, 19.0 and 19.0% in the treatment groups respectively. Biomass was the most sensitive parameter, with an EC 50 of 100 ppm ai; the NOAEC for biomass (and all other endpoints) was 4.0 ppb ai. The study is scientifically

sound and satisfies the US EPA Guideline 123

- 2 for an aquatic nonvascular study. The study is classified as core.9 D304987 46209403 850. 4550

Anabae na flos

- aquae In a 96

- hour acute toxicity study, cultures of Anabaena flos

- aquae were exposed to Cyazofamid under static conditions at nominal concentrations 0 (negative and solvent controls), 0.010, 0.026, 0.064, 0.16, 0.40, and 1.0 ppm ai. The time weig ht mean measured

concentrations were <0.0021

- 0.0022 (<LOQ, negative and solvent controls), 0.0049. 0.011, 0.027, 0.050, 0.16, and 0.25 ppm ai. At the

0

– hour analytical determination, the three lowest treatment concentrations produced recoveries >7

- % of no minal. The three highest nominal concentration exceeded the limit of water solubility (0.108 mg/L) for cyazofamid technical All concentrations of test material declined substantially over the course of the study, presumably due to the photodegradation of cyazofamid and to the presence of the algae.

The 96

- hour cell density percent inhibitions were 32, 20, 14, 12, 25

and 37% in the treatment groups respectively. The 72

- hour biomass (areas under the growth curve) percent inhibitions were 55, 25, 1, 17, 26, and 22% in the treatment groups respectively. The 72

- hour growth rate percent inhibitions were

- 5,

- 6,

- 6,

- 7, 32 and 7% in the treatment groups respectively. No significant inhibitions were observed for cell densities, growth rates or biomass compared to the pooled control.

This toxicity study was scientifically sound but does not satisfy the

US EPA Guideline Subdivision J , 123

- 2 because of the uncertainty

associated with the actual test concentrations that algae were exposed to and thus the inability to derive accurate toxicity values. However, no significant effects were note d up to the limit of solubility. As a result, this study is classified as supplemental.

D304897 46209404 850.4500

Navicula pelliculosa In a 96 hour acute toxicity study, cultures of Navicula pelliculosa were exposed to cyazofamid under static conditions at nominal concentrations 0 (negative and so lvent controls), 0.010, 0.026, 0.064, 0.16, 0.40 and 1.0 ppm ai. The time

- weighted mean measured

concentra tions were <0.0019

- 0.0028 (<LOQ , negative and solvent controls), 0.0039, 0.0075, 0.014, 0.019, 0.0073 and 0.12 ppm ai. At

the 0

- hour analytical dete rmination, only the two lowest treatment concentrations produced recoveries >70% of nominal. Furthermore, the three highest nominal concentrations exceeded the limit of water solubility (0.108 mg/L) for cyazofamid technical. All

concentrations of test ma terial declined substantially over the course of the study, presumable due to the photodegradation of cyazofamid and to the presence of the algae.

The 96

- hour cell density percent inhibitions were

- 39,

- 38,

- 71,

- 118,

- 110, and

- 6% in the treatment groups respectively. The 72

- hour biomass (areas under the growth curve) percent inhibitions were 17,10 - 27,

- 27,

- 47,

- 24 and

- 19% in the treatment groups respectively. No significant inhibitions were observed for the cell densities or growth rates, compared to the pooled control. The biomass, areas under the growth curve, was significantly reduced in the 0.073 and 0.12 ppm ai treatment groups compared to the pooled control.

This toxicity study is not scientifically sound and does not satisfy the US EPA Guideline Subdivision J, 123

- 2 because of the uncertainty associated with the actual test concentrations that algae were exposed to and thus the inabilit y to derive accurate toxicity values. However, there were no effects and as a result, this study is classified as supplemental.

ISKBC Comment: Pages 29 & 30, Section 11. Preliminary Identification of Data Gaps, b. Effects,

Table 7

- Data Needs, Bee t oxicity (6 special studies)

The need for both Tier I and Tier II tests following the new pollinator guidance is stated, but the guidance document (Guidance for Assessing Pesticide Risks to Bees, June 19, 2014) specifically outlines a tiered approach that clearly delineates a progression from Tier I to Tier
II tests, based on the results from the Tier I tests and calculation of the resulting risk (Figure 1, Tiered Approach for Assessing Risk to Honey Bees from Foliar Spray Applications, page 9 of guidance document). It seems premature to recommend conducting both Tier I and Tier II tests simultaneously, when the need for Tier II studies may be obviated, particularly since cyazofamid has been shown to be non

- toxic to bees. The more appropriate course of ac tion, would appear to be requiring the necessary Tier I tests only, followed by an evaluation according to the new guidelines before requiring the higher tier testing, in this case, the Tier II

field and semi

- field honeybee and pollen/nectar studies. Sinc e cyazofamid has been shown to

be non

- toxic to bees and since it is not a systemic product and thus will not move within the plant, we request waivers for all of the Tier II studies.

EFED Response: The EFED pollinator guidance outlines the process for completing study requirements and explains the requirement of the Tier II studies will be pending based on the results of acceptable

Tier I tests.

ISKBC Comment: Pages 31, Section 11. Preliminary Identific ation of Data Gaps, b. Effects,

Table 8

- Data Needs, invertebrate chronic and fish ELS (OCSPP 850.1300; 1400)

We have an o ctanol/water partition coefficient study resulting in a low Log Pow of 1.04 and an

aero bic soil metabolism study for CTCA that shows that CTCA is extensively mineralized in

soils with DT 50 values at 20°C of 21 to 33 days and DT 90 values of 69

- 109 days. Both of these

studies were required by Canada and were accepted by PMRA. We will submit them to the Agency for review. Since the acute LC 50 toxicity value for fish (rainbow trout) showed that CTCA was more than 100 times less toxic then parent, (>92.2 ppm vs. 0.56 ppm) these two studies should not be necessary. In addition, using this infor mation in ECOSAR, the resulting11 freshwater chronic NOEC value for invertebrates (0.184 ppm) is at least 10 times less toxic

than parent. The resulting ECOSAR NOEC value for fish (0.237 ppm) is in the same range as seen in the studies conducted using the pa rent. Thus, we request a waiver from conducting these

two studies for on CTCA.

EFED Response: The submitted studies are still in review with EFED thus we cannot specifically

comment at this time.