Risks of Diuron Use to Federally Threatened California Red-legged Frog (Rana aurora draytonii) Pesticide Effects Determination Environmental Fate and Effects Division Office of Pesticide Programs Washington, D.C. 20460 February 18, 2009 ------- Primary Authors: Ron Dean, Biologist Tiffany Mason, Environmental Engineer Bill Shaughnessy, PhD, Environmental Scientist Environmental Risk Branch II Environmental Fate and Effects Division (7507C) Secondary Review: William P. Eckel, PhD, Senior Scientist Environmental Risk Branch II Environmental Fate and Effects Division (7507P) Jean Holmes, Senior Scientist Environmental Risk Branch II Environmental Fate and Effects Division (7507P) Branch Chief, Environmental Risk Branch II: Tom Bailey, PhD, Branch Chief Environmental Fate and Effects Division (7507P) i ------- Table of Contents 1.0 Executive Summary 1 2.0 Problem Formulation 11 2.1 Purpose 11 2.2 Scope 13 2.3 Previous Assessments 14 2.4 Stressor Source and Distribution 15 2.4.1 Environmental Fate Properties 15 2.4.2 Environmental Transport Mechanisms 17 2.4.3 Mechanism of Action 21 2.4.4 Use Characterization 21 2.5 Assessed Species 25 2.5.1 Distribution 25 2.5.2 Reproduction 28 2.5.3 Diet 28 2.5.4 Habitat 29 2.6 Designated Critical Habitat 30 2.7 Action Area 32 2.8 Assessment Endpoints and Measures of Ecological Effect 35 2.8.1. Assessment Endpoints for the CRLF 36 2.8.2 Assessment Endpoints for Designated Critical Habitat 37 2.9 Conceptual Model 40 2.9.1 Risk Hypotheses 40 2.9.2 Diagram 40 2.10 Analysis Plan 44 2.10.1 Measures to Evaluate the Risk Hypothesis and Conceptual Model 44 2.10.1.1 Measures of Exposure 44 2.10.1.2 Measures of Effect 46 2.10.1.3 Integration of Exposure and Effects 47 2.10.2 Data Limitations 48 3.0 Exposure Assessment 48 3.1 Label Application Rates and Intervals 49 3.2 Aquatic Exposure Assessment 49 3.2.1 Modeling Approach 49 3.2.2 Model Inputs 49 3.2.3 Results 50 3.2.4 Existing Monitoring Data 52 3.2.6 Downstream Dilution Analysis 53 3.3 Terrestrial Animal Exposure Assessment 54 3.4 Terrestrial Plant Exposure Assessment 57 3.4.1 Spray Drift Buffer Analysis 59 4.0 Effects Assessment 61 4.1 Toxicity of Diuron to Aquatic Organisms 63 4.1.1 Toxicity to Freshwater Fish 65 4.1.2 Toxicity to Freshwater Invertebrates 65 li ------- 4.1.3 Toxicity to Aquatic Plants 65 4.2 Toxicity of Diuron to Terrestrial Organisms 66 4.2.1 Toxicity to Birds 67 4.2.2 Toxicity to Mammals 68 4.2.3 Toxicity to Terrestrial Invertebrates 68 4.2.4 Toxicity to Terrestrial Plants 68 4.3 Use of Probit Slope Response Relationship to Provide Information on the Endangered Species Levels of Concern 69 4.4 Incident Database Review 70 5.0 Risk Characterization 70 5.1 Risk Estimation 70 5.1.1 Exposures in the Aquatic Habitat 71 5.1.1.1 Direct Effects to Aquatic-Phase CRLF 71 5.1.1.2 Indirect Effects to Aquatic-Phase CRLF via Reduction in Prey (non-vascular aquatic plants, aquatic invertebrates, fish, and frogs) 79 5.1.1.3 Indirect Effects to CRLF via Reduction in Habitat and Primary Productivity (Freshwater Aquatic Plants) 86 Table 5.4 Summary of Acute RQs Used to Estimate Indirect Effects to the CRLF via Effects to Vascular Aquatic Plants (habitat of aquatic-phase CRLF)a 86 5.1.2 Exposures in the Terrestrial Habitat 89 5.1.2.1 Direct Effects to Terrestrial-phase CRLF 89 5.1.2.2 Indirect Effects to Terrestrial-Phase CRLF via Reduction in Prey (terrestrial invertebrates, mammals, and frogs) 91 5.1.2.3 Indirect Effects to CRLF via Reduction in Terrestrial Plant Community (Riparian and Upland Habitat) 93 5.1.3 Primary Constituent Elements of Designated Critical Habitat 94 5.1.3.1 Aquatic-Phase (Aquatic Breeding Habitat and Aquatic Non-Breeding Habitat) 94 5.1.3.2 Terrestrial-Phase (Upland Habitat and Dispersal Habitat) 95 5.2 Risk Description 96 5.2.1 Direct Effects 100 5.2.1.1 Aquatic-Phase CRLF 100 5.2.1.2 Terrestrial-Phase CRLF 101 5.2.2 Indirect Effects (via Reductions in Prey Base) 104 5.2.2.1 Algae (non-vascular plants) 104 5.2.2.2 Aquatic Invertebrates 105 5.2.2.3 Fish and Aquatic-phase Frogs 106 5.2.2.4 Terrestrial Invertebrates 106 5.2.2.5 Mammals 107 5.2.2.6 Terrestrial-phase Amphibians 107 5.2.3 Indirect Effects (via Habitat Effects) 108 5.2.3.1 Aquatic Plants (Vascular and Non-vascular) 108 5.2.3.2 Terrestrial Plants 108 5.2.4 Modification to Designated Critical Habitat 109 5.2.4.1 Aquatic-Phase PCEs 109 5.2.4.2 Terrestrial-Phase PCEs 110 in ------- 6.0 Uncertainties Ill 6.1. Exposure Assessment Uncertainties Ill 6.1.1 Maximum Use Scenario 111 6.1.2 Aquatic Exposure Modeling of Diuron 112 6.1.3 Action Area Uncertainties 113 6.1.4 Usage Uncertainties 114 6.1.5 Terrestrial Exposure Modeling of Diuron 114 6.2 Effects Assessment Uncertainties 115 6.2.1 Age Class and Sensitivity of Effects Thresholds 115 6.2.2 Use of Surrogate Species Effects Data 116 6.2.3 Sublethal Effects 117 6.2.4 Location of Wildlife Species 117 7.0 Risk Conclusions 117 8.0 References 122 Appendices Appendix A Multi-ai Product Analysis Appendix B Chemical Structure of Parent and Degradates Appendix C Use Rates and Methods Appendix D CDPR PUR Usage Data Appendix E RQ Method and LOCs Appendix F Spatial Summary of Diuron Appendix G PRZM/EXAMS Output Appendix H T-REX Example Output Appendix I TerrPlant Example Output Appendix J Bibliography of ECOTOX Open Literature Not Evaluated Appendix K Accepted ECOTOX Data Table Spreadsheet Appendix L Ecological Effects Data Appendix M Diuron Incidents Appendix N T-Herps Example Output Attachment I. Status and Life History of the California Red-legged Frog Attachment II. Baseline Status and Cumulative Effects for the California Red-legged Frog iv ------- List of Tables Table 1.1 Effects Determination Summary for Effects of Diuron on the CRLF 5 Table 1.2 Effects Determination Summary for the Critical Habitat Impact Analysis 6 Table 1.3 Diuron Use-specific Direct Effects Determinations1 for the CRLF 7 Table 1.4 Diuron Use-specific Indirect Effects Determinations1 Based on Effects to Prey8 Table 2.1 Summary of Diuron Half-lives 17 Table 2.2 Active Use Sites for Diuron as Applicable to California 24 Table 3.1 Summary of PRZM/EXAMS Environmental Fate Data Used for Aquatic Exposure Inputs for Diuron Endangered Species Assessment for the CRLF 50 Table 3.2 Table Aquatic EECs (ppb) for Diuron Uses in California 51 Table 3.3 T-REX Model Input Parameters 54 Table 3.4 Upper-bound Kenega Nomogram EECs for Dietary- and Dose-based Exposures of the CRLF and its Prey to Diuron 55 Table 3.5 EECs (ppm) for Indirect Effects to the Terrestrial-Phase CRLF via Effects to Terrestrial Invertebrate Prey Items 56 Table 3.6 TerrPlant Inputs and Resulting EECs for Plants Inhabiting Dry and Semi- aquatic Areas Exposed to diuron via Runoff and Drift 58 Table 3.7 Summary of AgDRIFT Predicted Spray Drift Buffer for Terrestrial Plants ... 60 Table 4.1 Freshwater Aquatic Toxicity Profile for Diuron 63 Table 4.2 Categories of Acute Toxicity for Aquatic Organisms 64 Table 4.3 Terrestrial Toxicity Profile for Diuron 66 Table 4.4 Categories of Acute Toxicity for Avian and Mammalian Studies 67 Table 4.5 Terrestrial Plant Toxicity Profile for Diuron 69 Table 5.1. Summary of Direct Effect RQs for the Aquatic-phase CRLF 72 Table 5.2 Summary of Acute RQs Used to Estimate Indirect Effects to the CRLF via Effects to Non-Vascular Aquatic Plants (diet of CRLF in tadpole life stage and habitat of aquatic-phase CRLF)* 80 Table 5.3 Summary of Acute and Chronic RQs Used to Estimate Indirect Effects to the CRLF via Direct Effects on Aquatic Invertebrates as Dietary Food Items (prey of CRLF juveniles and adults in aquatic habitats)* 83 Table 5.4 Summary of Acute RQs Used to Estimate Indirect Effects to the CRLF via Effects to Vascular Aquatic Plants (habitat of aquatic-phase CRLF)a 86 Table 5.5 Summary of Acute RQs Used to Estimate Direct Effects to the Terrestrial- phase CRLF 90 Table 5.6 Summary of Acute and Chronic RQs* Used to Estimate Indirect Effects to the Terrestrial-phase CRLF via Direct Effects on Small Mammals as Dietary Food Items .. 92 Table 5.7 RQs* : for Non-Target Plants Inhabiting Dry and Semi-Aquatic Areas Exposed to Diuron via Runoff and Drift 93 Table 5.8 Risk Estimation Summary for Diuron - Direct and Indirect Effects to CRLF . 97 Table 5.9 Risk Estimation Summary for Diuron - PCEs of Designated Critical Habitat for the CRLF 98 Table 5.10. Upper-bound Kenega Nomogram T-HERPS EECs (mg/kg-diet) for Dietary- based Exposures of the CRLF and its Prey to Diuron1 102 v ------- Table 5.11. Revised Acute Dietary-based RQs for CRLF consuming different food items (RQs calculated using T-HERPS)* 103 Table 7.1 Effects Determination Summary for Effects of Diuron on the CRLF 119 Table 7.2 Effects Determination Summary for the Critical Habitat Impact Analysis... 120 vi ------- List of Figures Figure 2.1 Diuron Agricultural Use in Total Pounds per County 23 Figure 2.2 Recovery Unit, Core Area, Critical Habitat, and Occurrence Designations for CRI.I 27 Figure 2.3 - CRLF Reproductive Events by Month 28 Figure 2.4 Initial area of concern, or "footprint" of potential use, for diuron 34 Figure 2.5 Conceptual Model for Aquatic-Phase of the CRLF 41 Figure 2.6 Conceptual Model for Terrestrial-Phase of the CRLF 42 Figure 2.7 Conceptual Model for Pesticide Effects on Aquatic Component of CRLF Critical Habitat 43 Figure 2.8 Conceptual Model for Pesticide Effects on Terrestrial Component of CRLF Critical Habitat 44 vii ------- 1.0 Executive Summary The purpose of this assessment is to evaluate potential direct and indirect effects on the California red-legged frog (Rana aurora draytonii) (CRLF), from FIFRA regulatory actions regarding use of diuron on agricultural and non-agricultural sites. In addition, this assessment evaluates whether these actions can be expected to result in modification of the species' designated critical habitat. This assessment was completed in accordance with the U.S. Fish and Wildlife Service (USFWS) and National Marine Fisheries Service (NMFS) Endangered Species Consultation Handbook (USFWS/NMFS, 1998) and procedures outlined in the Agency's Overview Document (U.S. EPA, 2004). The CRLF was listed as a threatened species by USFWS in 1996. The species is endemic to California and Baja California (Mexico) and inhabits both coastal and interior mountain ranges. A total of 243 streams or drainages are believed to be currently occupied by the species, with the greatest numbers in Monterey, San Luis Obispo, and Santa Barbara counties (USFWS, 1996) in California. Diuron (N-(3,4-dichlorophenyl)-N,N-dimethylurea) is a systemic substituted phenylurea herbicide. In the U.S. diuron is used on a variety of fruit and nut crops, grains, cotton, corn, sorghum, mint, gum, asparagus, sugarcane, seed crops, coffee, hay, cut flowers, and for fallow, idle cropland use, and impervious surfaces such as paved areas. It may be used in irrigation and drainage systems when water is not present. Diuron is available in wettable powder, flowable, liquid suspension, and soluble concentrate formulations for use in California. Technical diuron is a white, crystalline, and odorless solid. Diuron is registered for pre- and post emergent control using ground and aerial equipment. Diuron is often applied in combination with other herbicides such as bromacil, hexazinone, paraquat, thiadiazuron, imazapyr, monosodium, sodium chlorate, sodium metaborate, and copper sulfate (U.S. EPA, 2004a). This assessment evaluates only those registered uses which allow use in California. Diuron is easily taken up by plants and rapidly translocated. Diuron primarily functions by inhibiting the Hill reaction in photosynthesis, limiting the production of high-energy compounds such as adenosine triphosphate (ATP) used for various metabolic processes. Diuron is generally considered to be both mobile and persistent. It is relatively stable in neutral water. In addition to aquatic photolysis, microbial degradation is the primary factor in the degradation of diuron in aquatic environments. Diuron is not expected to concentrate in aquatic organisms. Diuron is regulated as a known ground water contaminant in California, where it has been detected in the 2 to 3 ppb range (online at http://www.cdpr.ca.gov). Because diuron has a low Henry's law constant and a low vapor pressure, volatilization is considered insignificant. At sufficient levels diuron residues in soil are toxic to plants. Residue levels are lower in soils with low organic content. Residue half-lives are from one month to one year for the parent, five months for a major degradate , and one month for a minor degradate. 1 ------- Degradate formation is summarized below. Some pineapple fields contained residues three years after the last application. Diuron degrades in the environment to four major (>10% of the applied parent) and four minor (<10% of the applied parent) metabolites and impurities. The major metabolites are: carbon dioxide (C02), N-(3,4-dichlorophenyl)-N-methylurea (DCPMU), N'-(3- chlorophenyl)-N,N-dimethylurea (MCPDMU), and l,l-dimethyl-3-phenylurea (PDMU). The minor metabolites include: 4-dichlorophenylurea (DCPU); 3,4-dichloroaniline (3,4- DCA), N-(3-chlorophenyl)-N-methylurea (CPMU), and 3,3',4,4'-tetrachlorobenzene (TCAB). There were no acceptable toxicity studies on the major degradates identified in the ECOTOX database. As a result, they will be assigned the same toxicity as the parent for purposes of this assessment. Because the concentrations of parent diuron exceed levels of concern by themselves (both in modeling and monitoring data), the non-inclusion of degradates does not cause a different outcome for the "may affect" determination. Therefore, this assessment only addresses ecological risks associated with exposures to the parent compound. Likewise, the minor degradates, 3,4-DCA and TCAB, will not be assessed further. 3,4 DCA is not a significant residue (< 1%) in any metabolism or hydrolysis study, and TCAB is also a minor degradate (0.038 ppm) and is only found in the soil photolysis study which is a minor degradation route for diuron. Diuron contains two impurities from the manufacturing process, TCAB and 3,3',4,4'- tetrachloroazoxybenzene (TCAOB), both 'dioxin-like' substances. TCAB levels between 0.15 and 28 ppm have been found in diuron samples tested. TCAOB is present at lower levels. Since, these impurities have only been found at fairly low levels, they are not residues of concern for diuron and will not be evaluated in this assessment. Diuron is classified as a 'known/likely' carcinogen. EPA is required under the FFDCA, as amended by FQPA, to develop a screening program to determine whether certain substances (including all pesticide active and other ingredients) "may have an effect in humans that is similar to an effect produced by a naturally occurring estrogen, or other such endocrine effects as the Administrator may designate. " Following the recommendations of its Endocrine Disruptor Screening and Testing Advisory Committee (EDSTAC), EPA determined that there were scientific bases for including, as part of the program, androgen and thyroid hormone systems, in addition to the estrogen hormone system. EPA also adopted EDSTAC's recommendation that the Program include evaluations of potential effects in wildlife. When the appropriate screening and/or testing protocols being considered under the Agency's Endocrine Disrupter Screening Program (EDSP) have been developed and vetted, diuron may be subjected to additional screening and/or testing to better characterize effects related to endocrine disruption. As noted in the 2003 Registration Eligibility Decision (RED), diuron has been characterized as a "known/likely" carcinogen, based on urinary bladder carcinomas in both sexes of the Wistar rat, kidney carcinomas in the male rat (a rare tumor), and 2 ------- mammary gland carcinomas in the female NMRI mouse. Because of this, the spatial extent of the action area (i.e., the boundary where exposures and potential effects are less than the Agency's LOC) for diuron cannot be determined. Therefore, it is assumed that the action area encompasses the entire state of California, regardless of the spatial extent (i.e., initial area of concern or footprint) of the pesticide use(s). Since CRLFs exist within aquatic and terrestrial habitats, exposure of the CRLF, its prey and its habitats to diuron are assessed separately for the two habitats. Tier-II aquatic exposure models are used to estimate high-end exposures of diuron in aquatic habitats resulting from runoff and spray drift from different uses. Peak model-estimated environmental concentrations resulting from different diuron uses range from 3.76 to 4911 ppb for use on grasses grown for seed, and paved areas (impervious surfaces), respectively. These estimates are supplemented with analysis of available California surface water monitoring data from U. S. Geological Survey's National Water Quality Assessment (NAWQA) program and the California Department of Pesticide Regulation (CDPR). Particularly in the CDPR monitoring data, diuron is frequently detected and exceeds levels of concern with values up to 160 ppb. To estimate on-site diuron dietary exposures to the terrestrial-phase CRLF, and its potential prey resulting from uses involving diuron applications, the T-REX model is used for spray treatment. The AgDRIFT model is also used to estimate deposition of diuron on off-site terrestrial and aquatic habitats from spray drift. The TerrPlant model is used to estimate diuron exposures to terrestrial-phase CRLF habitat, including plants inhabiting semi-aquatic and dry areas, from site run-off and spray drift. The T-HERPS model is used to allow for further characterization of dietary exposures of terrestrial- phase CRLFs relative to birds. The effects determination assessment endpoints for the CRLF include direct toxic effects on the survival, reproduction, and growth of the CRLF itself. Direct effects to the CRLF in the aquatic habitat are based on toxicity information for freshwater fish, which are generally used as a surrogate for aquatic-phase amphibians. In the terrestrial habitat, direct effects are based on toxicity information for birds, which are used as a surrogate for terrestrial-phase amphibians. Given that the CRLFs prey items and designated critical habitat requirements in the aquatic habitat are dependant on the availability of freshwater aquatic invertebrates and aquatic plants, toxicity information for these taxonomic groups is also discussed. In the terrestrial habitat, indirect effects due to depletion of prey are assessed by considering effects to terrestrial insects, small terrestrial mammals, and frogs. Indirect effects due to modification of the terrestrial habitat are characterized by available data for terrestrial monocotyledonous (monocot) and dicotyledonous (dicot) plants. Risk quotients (RQs) are derived as estimates of potential risk. Acute and chronic RQs for animals and listed and non-listed plant RQs are compared to the Agency's levels of concern (LOCs) to identify instances where diuron use within the action area has the 3 ------- potential to adversely affect the CRLF via direct toxicity or indirectly based on effects to its food supply (i.e., freshwater invertebrates, algae, fish, frogs, terrestrial invertebrates, and mammals) or habitat (i.e., aquatic plants and terrestrial upland and riparian vegetation) or to modify its critical habitat. When RQs for each particular type of effect are below their respective LOCs, the pesticide is determined to have "no effect" on the CRLF or to modify its critical habitat. Where RQs exceed LOCs, a potential to cause adverse effects is identified, leading to a conclusion of "may affect" for the CRLF and a potential to modify its critical habitat. If a determination is made that use of diuron within the action area "may affect" the CRLF, additional information is considered to refine the potential for exposure and effects, and the best available information is used to distinguish those actions that "may affect, but are not likely to adversely affect" (NLAA) from those actions that are "likely to adversely affect" (LAA) the CRLF. Similarly for critical habitat additional information is considered to refine the potential for exposure and effects to distinguish those actions that do or do result in modification of its critical habitat. Based on the best available information, the Agency makes a May Affect, and Likely to Adversely Affect determination for diuron exposure to the CRLF based on direct and indirect effects to the aquatic- and terrestrial-phase CRLF. Further, registered uses of diuron are anticipated to result in modification to critical habitat of the CRLF. A summary of the risk conclusions and effects determinations for the CRLF and its critical habitat is presented in Table 1.1 and Table 1.2. Use-specific determinations for direct and indirect effects to the CRLF are provided in Table 1.3 and Table 1.4. Further information on the results of the effects determination is included as part of the Risk Description in Section 5.2. Given the LAA determination for the CRLF and potential modification of designated critical habitat, a description of the baseline status and cumulative effects for the CRLF is provided in Attachment II. 4 ------- Table 1.1 Effects Determination Summary for Effects of Diuron on the CRLF Assessment Endpoint Etfeets Determination 1 Basis for Determination Survival, growth, and/or reproduction of CRLF individuals LAA Potential for Direct Effects Aquatic-phase (Eggs, Larvae, and Adults) : The aquatic phase amphibian acute LOCs for Listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 12.28 (paved areas) to 0.06 (papaya and walnut). Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 131.85 (paved areas) to 1.26 (banana, plantain). Terrestrial-phase (Juveniles and Adults): Refined acute dietary-based RQs for CRLFs consuming large insects and small herbivore mammals exceed the acute listed species LOC (0.1) for all uses of diuron except Sorghum and Field Corn. The acute dietary-based RQs for CRLFs consuming large insects exceed the acute listed species LOC for Agricultural Rights-of-Way, Fencerows etc. No acute dietary based LOCs were exceeded for CLRF consuming small insectivore mammals and small terrestrial phase amphibians for any diuron use. LAA Potential for Indirect Effects Aquatic prey items, aquatic habitat, cover and/or primary productivity. LOCs for non-vascular plants are exceeded for all uses. The RQs range from 2046.25 (paved areas) to 1.57 (grasses grown for seed). Aquatic invertebrates acute LOCs are exceeded. The acute RQs that exceed the LOC range from 30.69 (paved areas) to 0.05 (cotton aerial application). Chronic LOCs are exceeded for several uses. RQs that exceed the Agency's LOC range from 21.21or paved areas to 1.02 for bermudagrass. RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 460.46 (paved areas) to 1.30 (hazelnut). Terrestrial prey items, riparian habitat. For small mammals, chronic dose based RQs exceed the Agency's LOC (1.0) for most uses (Table 5.6). The dose based chronic RQs that exceed the LOC range from 8.29 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 1.31 (walnut). The dietary based chronic RQs do not exceed the LOC for any uses. The RQs that exceed the acute dose based LOC range from 0.41 (Agricultural Rights- of-Way, Fencerows, Hedgerows, Airports, etc.) to 0.13 (Bermuda grass, Blackberry, Boysenberry, Spearmint). For terrestrial phase amphibians, the chronic LOC is exceeded for all uses of diuron and the acute LOCs are exceeded for all uses except sorghum. LOCs are exceeded for terrestrial riparian plants and for aquatic plants from exposure to diuron from runoff or spray drift. Alteration of riparian and vascular plants may result in alteration of temperature, turbidity, and oxygen content. RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 460.46 (paved areas) to 1.30 (hazelnut). 1 No effect (NE); May affect, but not likely to adversely affect (NLAA); May affect, likely to adversely affect (LAA) 5 ------- Table 1.2 Effects Determination Summary for the Critical Habitat Impact Analysis Assessment End point Effects Determination 1 Basis for Determination Modification of aquatic-phase PCE HM Due to aquatic vascular and terrestrial plant communities being reduced from all use sites, there is potential for alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond. These plant communities provide for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. In addition, there is potential for alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food. LOCs are exceeded for terrestrial riparian plants and for aquatic vascular plants from exposure to diuron from runoff or spray drift. LOCs for non-vascular plants are exceeded for all uses. Modification of terrestrial-phase PCE HM The use of diuron at all use sites may create the following modification of PCE: elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs, elimination and/or disturbance of dispersal habitat, reduction and/or modification of food sources for terrestrial phase juveniles and adults, and alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. The RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses of diuron in California. RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 460.46 (paved areas) to 1.30 (hazelnut). Use of diuron on most use sites will exceed acute and chronic LOCs for many prey food items of the CRLF. Food sources are reduced and the CRLF is therefore indirectly affected. The RQs for non-target terrestrial monocot and dicot plants inhabiting semi-aquatic and upland dry areas exceed the Agency's LOC (1.0) for all uses except for monocot plants exposed to sorghum applications (Table 5.7). These exceedances range from 300.00 (semi-aquatic plants exposed to non-agricultural aerial applications) to 0.19 (dicot plants inhabiting dry areas exposed to sorghum applications). Several diuron uses result in plant LOC exceedances from spray drift. These exceedances range from 12.00 (dicot plants exposed to field corn applications) to 1.13 (monocot plants exposed to cotton applications). 1 Habitat Modification (HM) or No effect (NE) 6 ------- Table 1.3 Diuron Use-specific Direct Effects Determinations1 for the CRLF I sols) Auiiiilie lliihiliil lerreslriiil lliihiliil Anile ( lironie Aeule (lironie- AGRICULTURAL RIGHTS-OF-WAY / FENCEROWS/ HEDGEROWS LAA LAA LAA LAA ALFALFA Ground NE NE LAA LAA ALFALFA Aerial NE NE LAA LAA APPLE NE NE LAA LAA ARTICHOKE LAA NE LAA LAA ASPARGUS LAA NE LAA LAA BANANA/PLANTAIN LAA LAA LAA LAA BLACKBERRY/BOYSENBERRY LAA LAA LAA LAA BLUEBERRY NE NE LAA LAA CITRUS NE NE LAA LAA CORN, FIELD NE NE NE LAA DEWBERRY NE NE LAA LAA FILBERT (HAZELNUT) NE NE LAA LAA GRAPE NE NE LAA LAA LOGANBERRY, RASPBERRY (BLACK/RED) NE NE LAA LAA OLIVE NE NE LAA LAA PAPAYA NE NE LAA LAA PEPPERMINT NE NE LAA LAA PEACH NE NE LAA LAA PEAR NE NE LAA LAA PECAN LAA NE LAA LAA SORGHUM NE NE NE LAA SPEARMINT LAA LAA LAA LAA WALNUT (ENGLISH/BLACK) LAA NE LAA LAA WHEAT Pre Harvest NE NE LAA LAA WHEAT Post Harvest NE NE LAA LAA COTTON Ground NE NE LAA LAA COTTON Aerial NE NE LAA LAA AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS LAA LAA LAA LAA BERMUDAGRASS LAA LAA LAA LAA GRASSES GROWN FOR SEED NE NE LAA LAA IRRIGATION SYSTEMS LAA LAA LAA LAA ORNAMENTAL HERBACEOUS PLANTS LAA NE LAA LAA NON-AGRICULTURAL RIGHTS OF WAY Ground LAA LAA LAA LAA NON-AGRICULTURAL RIGHTS OF WAY Aerial LAA LAA LAA LAA PAVED AREAS (PRIVATE ROADS/SIDEWALKS) LAA LAA LAA LAA UNCULTIVATED AGRICULTIRAL AREAS Ground LAA LAA LAA LAA UNCULTIVATED AGRICULTIRAL AREAS Aerial LAA LAA LAA LAA UNCULTIVATED NON-AGRICULTURAL AREAS LAA LAA LAA LAA 1 NE = No effect; MA/NLAA = May affect, but not likely to adversely affect; LAA = May affect and likely to adversely affect * No acceptable chronic data are available for amphibians or their avian surrogates by which to determine chronic direct risks to the CRLF, therefore an LAA determination is made in the absence of these data and because supporting information on linuron, a similar herbicide, exhibits chronic toxicity to these species. 7 ------- Table 1.4 Diuron Use-specific Indirect Effects Determinations1 Based on Effects to Prey l Sl(S) AI»;k- A(|ii;ilk' Im Auik- i'i'U'l>r;ik's (linink TiTivsiri.il liiM-i'k-hr.ik-s (Auilc) A(|ll;ilii-| illU Auik- )h;isi- fni»s llsli (linink 1'iTivsiriiil Auilc -pliiisi- fni»s ('linink'" Sni:iII \ Auik- l.imm.ils ('linink' AGRICULTURAL RIGIITS-OF- WAY/FENCEROWS/ HEDGEROWS LA A LAA LAA NLAA LAA LAA LAA LAA LAA LAA ALFALFA Ground LA A LAA NE NLAA NE NE LAA LAA NE LAA ALFALFA Aerial LA A LAA NE NLAA NE NE LAA LAA NE LAA APPLE LA A LAA NE NLAA NE NE LAA LAA NE LAA ARTICHOKE LA A LAA NE NLAA LAA NE LAA LAA NE LAA ASPARGUS LA A LAA NE NLAA LAA NE LAA LAA NE LAA BANANA/PLANTAIN LA A LAA NE NLAA LAA LAA LAA LAA LAA LAA BLACKBERRY/ BOYSENBERRY LA A LAA NE NLAA LAA LAA LAA LAA LAA LAA BLUEBERRY LA A NE NE NLAA NE NE LAA LAA NE NE CITRUS LA A NE NE NLAA NE NE LAA LAA NE LAA CORN, FIELD LA A NE NE NLAA NE NE NE LAA NE NE DEWBERRY LA A LAA NE NLAA NE NE LAA LAA NE LAA FILBERT (HAZELNUT) LA A LAA NE NLAA NE NE LAA LAA NE NE GRAPE LA A LAA NE NLAA NE NE LAA LAA NE LAA LOGANBERRY, RASPBERRY (BLACK/RED) LA A NE NE NLAA NE NE LAA LAA NE NE OLIVE LA A NE NE NLAA NE NE LAA LAA NE NE PAPAYA LA A LAA NE NLAA LAA NE LAA LAA NE LAA PEPPERMINT LA A NE NE NLAA NE NE LAA LAA NE LAA PEACH LA A LAA NE NLAA NE NE LAA LAA NE LAA PEAR LA A LAA NE NLAA NE NE LAA LAA NE NE PECAN LA A LAA NE NLAA NE NE LAA LAA NE LAA SORGHUM LA A LAA NE NLAA NE NE NE LAA NE NE SPEARMINT LA A LAA NE NLAA LAA LAA LAA LAA LAA LAA WALNUT (ENGLISH/BLACK) LA A LAA NE NLAA NE NE LAA LAA NE LAA WHEAT Pre Harvest LA A LAA NE NLAA NE NE LAA LAA NE NE 8 ------- I Sl(S) AI»;k- A(|ii;ilk' Im .Willi- iTU-l>r;ik-s ('liriuik TiTivsiri.il ln\iTli-l>i;ili-s (.Willi-) A(|ll;ilii-| ;ill( .Willi- >h;isi- fni»s llsli Ch in nil Ti-nvsirkil .Willi- -ph.isi- fni»s ('liniiik'-'- Sm:iII \ .Willi- l.imm.ils Ch milk WHEAT Post Harvest LAA LAA NE NLAA NE NE LAA LAA NE NE COTTON Ground LAA NE NE NLAA NE NE LAA LAA NE LAA COTTON Aerial LAA LAA NE NLAA NE NE LAA LAA NE LAA AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA BERMUDAGRASS LAA LAA LAA NLAA NE NE LAA LAA LAA LAA GRASSES GROWN FOR SEED LAA NE NE NLAA NE NE LAA LAA NE NE IRRIGATION SYSTEMS LAA LAA NE NLAA LAA LAA LAA LAA LAA LAA ORNAMENTAL HERBACEOUS PLANTS LAA LAA NE NLAA LAA LAA LAA LAA NE LAA NON-AGRICULTURAL RIGHTS OF WAY Ground LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA NON-AGRICULTURAL RIGHTS OF WAY Aerial LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA PAVED AREAS (PRIVATE ROADS/SIDEWALKS) LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA UNCULTIVATED AGRICULTIRAL AREAS Ground LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA UNCULTIVATED AGRICULTIRAL AREAS Aerial LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA UNCULTIVATED NON- AGRICULTURAL AREAS LAA LAA LAA NLAA LAA LAA LAA LAA LAA LAA 1 NE = No effect; NLAA = May affect, but not likely to adversely affect; LAA = May affect and likely to adversely affect * No acceptable chronic data are available for amphibians or their avian surrogates by which to determine chronic direct risks to the CRLF, therefore an LAA determination is made in the absence of these data and because linuron, a similar herbicide, exhibits chronic toxicity to these species in the range of modeled diuron surface water levels. 9 ------- Based on the conclusions of this assessment, a formal consultation with the U. S. Fish and Wildlife Service under Section 7 of the Endangered Species Act should be initiated. When evaluating the significance of this risk assessment's direct/indirect and adverse habitat modification effects determinations, it is important to note that pesticide exposures and predicted risks to the species and its resources (i.e., food and habitat) are not expected to be uniform across the action area. In fact, given the assumptions of drift and downstream transport (i.e., attenuation with distance), pesticide exposure and associated risks to the species and its resources are expected to decrease with increasing distance away from the treated field or site of application. Evaluation of the implication of this non-uniform distribution of risk to the species would require information and assessment techniques that are not currently available. Examples of such information and methodology required for this type of analysis would include the following: • Enhanced information on the density and distribution of CRLF life stages within specific recovery units and/or designated critical habitat within the action area. This information would allow for quantitative extrapolation of the present risk assessment's predictions of individual effects to the proportion of the population extant within geographical areas where those effects are predicted. Furthermore, such population information would allow for a more comprehensive evaluation of the significance of potential resource impairment to individuals of the species. • Quantitative information on prey base requirements for individual aquatic- and terrestrial-phase frogs. While existing information provides a preliminary picture of the types of food sources utilized by the frog, it does not establish minimal requirements to sustain healthy individuals at varying life stages. Such information could be used to establish biologically relevant thresholds of effects on the prey base, and ultimately establish geographical limits to those effects. This information could be used together with the density data discussed above to characterize the likelihood of adverse effects to individuals. • Information on population responses of prey base organisms to the pesticide. Currently, methodologies are limited to predicting exposures and likely levels of direct mortality, growth or reproductive impairment immediately following exposure to the pesticide. The degree to which repeated exposure events and the inherent demographic characteristics of the prey population play into the extent to which prey resources may recover is not predictable. An enhanced understanding of long-term prey responses to pesticide exposure would allow for a more refined determination of the magnitude and duration of resource impairment, and together with the information described above, a more complete prediction of effects to individual frogs and potential modification to critical habitat. 10 ------- 2.0 Problem Formulation Problem formulation provides a strategic framework for the risk assessment. By identifying the important components of the problem, it focuses the assessment on the most relevant life history stages, habitat components, chemical properties, exposure routes, and endpoints. The structure of this risk assessment is based on guidance contained in U.S. EPA's Guidance for Ecological Risk Assessment (U.S. EPA 1998), the Services' Endangered Species Consultation Handbook (USFWS/NMFS 1998) and is consistent with procedures and methodology outlined in the Overview Document (U.S. EPA 2004) and reviewed by the U.S. Fish and Wildlife Service and National Marine Fisheries Service (USFWS/NMFS 2004). 2.1 Purpose The purpose of this endangered species assessment is to evaluate potential direct and indirect effects on individuals of the federally threatened California red-legged frog (Rana aurora draytonii) (CRLF) arising from FIFRA regulatory actions regarding use of diuron on terrestrial food, feed, forestry, non-food, and outdoor residential sites. In addition, this assessment evaluates whether diuron use on these crops is expected to result in modification of the species' designated critical habitat. This ecological risk assessment has been prepared consistent with a settlement agreement in the case Center for Biological Diversity (CBD) vs. EPA etal. (Case No. 02-1580-JSW (JL)) settlement entered in Federal District Court for the Northern District of California on October 20, 2006. Diuron is a broad-spectrum residual herbicide registered for pre-emergent and post- emergent control of both broadleaf and annual grassy weeds. When diuron is used on pre-emergent weeds, it allows seeds to germinate normally, but causes them to lose their green color, after which they soon die of starvation (Ferrell et al., 2004). In the U.S. diuron is used on a variety of fruit and nut crops, grains, cotton, corn, sorghum, mint, gum, asparagus, sugarcane, seed crops, coffee, hay, cut flowers, and for fallow and idle cropland use. It may be used in irrigation and drainage systems when water is not present. It is also used to control weeds on hard surfaces, such as, roads, railway tracks, and paths (at around 3 kg/ha), and to control weeds in crops, such as, pear and apple trees, forestry, ornamental trees and shrubs, pineapples, sugar cane, cotton, alfalfa and wheat (at lower rates of around 1.8 kg/ha). It can be used for both pre-emergent and knockdown weed control. Its use in some locations is becoming limited due to the development of resistant weed species. In some products it is combined with other active ingredients such as glyphosate, bromacil, hexazinone, amitrole and 2,4-D. In some countries diuron is also registered for aquatic weed control, as a cotton defoliant, and for use in home aquaria and fish ponds. It is used as a booster biocide in antifouling paints where its activity enhances the efficacy of copper in these products. 11 ------- In this assessment, direct and indirect effects to the CRLF and potential modification to its designated critical habitat are evaluated in accordance with the methods described in the Agency's Overview Document (U.S. EPA 2004). Screening level methods include use of standard models such as PRZM-EXAMS, T-REX, TerrPlant, and AgDRIFT, all of which are described at length in the Overview Document. Additional refinements include use of the T-HERPS model. Use of such information is consistent with the methodology described in the Overview Document (U.S. EPA 2004), which specifies that "the assessment process may, on a case-by-case basis, incorporate additional methods, models, and lines of evidence that EPA finds technically appropriate for risk management objectives" (Section V, page 31 of U.S. EPA 2004). In accordance with the Overview Document, provisions of the ESA, and the Services' Endangered Species Consultation Handbook, the assessment of effects associated with registrations of diuron is based on an action area. The action area is the area directly or indirectly affected by the federal action, as indicated by the exceedance of the Agency's LOCs. It is acknowledged that the action area for a national-level FIFRA regulatory decision associated with a use of diuron may potentially involve numerous areas throughout the United States and its Territories. However, for the purposes of this assessment, attention will be focused on relevant sections of the action area including those geographic areas associated with locations of the CRLF and its designated critical habitat within the state of California. As part of the "effects determination," one of the following three conclusions will be reached regarding the potential use of diuron in accordance with current labels: • "No effect"; • "May affect, but not likely to adversely affect"; or • "May affect and likely to adversely affect". Designated critical habitat identifies specific areas that have the physical and biological features, (known as primary constituent elements or PCEs) essential to the conservation of the listed species. The PCEs for CRLFs are aquatic and upland areas where suitable breeding and non-breeding aquatic habitat is located, interspersed with upland foraging and dispersal habitat. If the results of initial screening-level assessment methods show no direct or indirect effects (no LOC exceedances) upon individual CRLFs or upon the PCEs of the species' designated critical habitat, a "no effect" determination is made for use of diuron as it relates to this species and its designated critical habitat. If, however, potential direct or indirect effects to individual CRLFs are anticipated or effects may impact the PCEs of the CRLFs designated critical habitat, a preliminary "may affect" determination is made for the FIFRA regulatory action regarding diuron. If a determination is made that use of diuron within the action area(s) associated with the CRLF "may affect" this species or its designated critical habitat, additional information is considered to refine the potential for exposure and for effects to the CRLF and other taxonomic groups upon which these species depend (e.g., aquatic and terrestrial 12 ------- vertebrates and invertebrates, aquatic plants, riparian vegetation, etc.). Additional information, including spatial analysis (to determine the geographical proximity of CRLF habitat and diuron use sites) and further evaluation of the potential impact of diuron on the PCEs is also used to determine whether modification of designated critical habitat may occur. Based on the refined information, the Agency uses the best available information to distinguish those actions that "may affect, but are not likely to adversely affect" from those actions that "may affect and are likely to adversely affect" the CRLF or the PCEs of its designated critical habitat. This information is presented as part of the Risk Characterization in Section 5 of this document. The Agency believes that the analysis of direct and indirect effects to listed species provides the basis for an analysis of potential effects on the designated critical habitat. Because diuron is expected to directly impact living organisms within the action area (defined in Section 2.7), critical habitat analysis for diuron is limited in a practical sense to those PCEs of critical habitat that are biological or that can be reasonably linked to biologically mediated processes (i.e., the biological resource requirements for the listed species associated with the critical habitat or important physical aspects of the habitat that may be reasonably influenced through biological processes). Activities that may modify critical habitat are those that alter the PCEs and appreciably diminish the value of the habitat. Evaluation of actions related to use of diuron that may alter the PCEs of the CRLFs critical habitat form the basis of the critical habitat impact analysis. Actions that may affect the CRLFs designated critical habitat have been identified by the Services and are discussed further in Section 2.6. 2.2 Scope Diuron is registered in California for a variety of agricultural and non-agricultural uses (see Table 2.2). The end result of the EPA pesticide registration process {i.e., the FIFRA regulatory action) is an approved product label. The label is a legal document that stipulates how and where a given pesticide may be used. Product labels (also known as end-use labels) describe the formulation type {e.g., liquid or granular), acceptable methods of application, approved use sites, and any restrictions on how applications may be conducted. Thus, the use or potential use of diuron in accordance with the approved product labels for California is "the action" relevant to this ecological risk assessment. Although current registrations of diuron allow for use nationwide, this ecological risk assessment and effects determination addresses currently registered uses of diuron in portions of the action area that are reasonably assumed to be biologically relevant to the CRLF and its designated critical habitat. Further discussion of the action area for the CRLF and its critical habitat is provided in Section 2.7. The major route of dissipation for diuron is microbial degradation in water. Here the major metabolism degradate is N'-(3-chlorophenyl)-N,N-dimethylurea (MCPDMU). Diuron also degrades through photolysis in both water and soil; here the major degradates 13 ------- are C02 and N'-(3,4-dichlorophenyl)-N-methylurea (DCPMU), respectively. A more detailed discussion of the major and minor degradates formed in the decomposition of diuron is presented in Section 2.4. The Agency does not routinely include, in its risk assessments, an evaluation of mixtures of active ingredients, either those mixtures of multiple active ingredients in product formulations or those in the applicator's tank. In the case of the product formulations of active ingredients (that is, a registered product containing more than one active ingredient), each active ingredient is subject to an individual risk assessment for regulatory decision regarding the active ingredient on a particular use site. If effects data are available for a formulated product containing more than one active ingredient, they may be used qualitatively or quantitatively in accordance with the Agency's Overview Document and the Services' Evaluation Memorandum (U.S., EPA 2004; USFWS/NMFS 2004). Diuron has registered products that contain multiple active ingredients. However, there are no available data on mixtures containing diuron in the open literature. There are no product LD50 values, with associated 95% Confidence Intervals (CIs) available for diuron among the data submitted to the Agency. The assessment will be based on the toxicity of a single active ingredient of diuron. Analysis of the available open literature and acute mammalian toxicity data for multiple active ingredient products relative to the single active ingredient is provided in Appendix A. As discussed in USEPA (2000) a quantitative component-based evaluation of mixture toxicity requires data of appropriate quality for each component of a mixture. In this mixture evaluation, an LD50 with associated 95% CI is needed for the formulated product. The same quality of data is also required for each component of the mixture. Given that the formulated products for diuron do not have LD50 data available it is not possible to undertake a quantitative or qualitative analysis for potential interactive effects. However, because the active ingredients are not expected to have similar mechanisms of action, metabolites, or toxicokinetic behavior, it is reasonable to conclude that an assumption of dose-addition would be inappropriate. Consequently, an assessment based on the toxicity of diuron is the only reasonable approach that employs the available data to address the potential acute risks of the formulated products. 2.3 Previous Assessments Diuron has been registered in the United States since 1967 for use as an herbicide, mildewcide and algaecide. A Registration Standard, titled "Guidance for the Reregi strati on of Pesticide Products Containing Diuron as the Active Ingredient" was released in 1983. The Registration Standard involved a thorough review of the scientific data base underlying pesticide registrations and an identification of essential but missing studies which may not have been required when the product was initially registered or studies that were considered insufficient. Subsequent Data Call-Ins (DCIs) were issued in 1990, and 1995 for diuron. 14 ------- In 2003, the Office of Pesticide Programs (OPP) requested the initiation of ESA section 7(a) (2) consultation regarding the potential impact of diuron on the Evolutionary Significant Units (ESU) of the Pacific salmon and steelhead. OPP determined that the use of diuron on certain crops (peaches, walnuts, filberts) may affect 10 ESUs at application rates above 3.2 lb ai/A, may affect 7 ESUs at application rates above 2.2 lb ai/A, and will have no effect on 10 ESUs at any labeled agricultural rate. The use of diuron for non-crop sites, especially rights-of-way, may affect 25 ESUs, may affect but is not likely to adversely affect one ESU, and will have no effect on one ESU. To mitigate risks of concern posed by the use of diuron, EPA in its Reregi strati on Eligibility Decision (RED) for diuron, dated September 2003, noted a number of label amendments identified by EPA or agreed to by the registrant(s) to address the worker, residential and ecological concerns; these included: • All wettable powder products will be voluntarily canceled. • Reduction in application rate and increased treatment intervals, and limit on the number of applications for some crops. • Use of the backpack sprayer is prohibited. • Implement use of PPE and engineering controls for some workers. • Eliminate aerial applications except for rights-of-way, alfalfa, cotton, winter barley, winter wheat, sugarcane, and grass seed crops. • Implement best management practices to reduce spray drift. 2.4 Stressor Source and Distribution 2.4.1 Environmental Fate Properties The environmental fate database for diuron is essentially complete. Diuron is stable in neutral media at normal temperatures, and is hydrolyzed by acid and alkalis. It is stable towards oxidation and moisture under normal conditions and decomposes at 180-190° C (Helliwell et al., 1998). Diuron is mobile and has the potential to leach to groundwater and to contaminate surface waters. An upgradable adsorption/desorption/leaching study (MRID 44490501) showed that diuron has an average Koc value of 920 and Freundlich Kads values (7.9-28). Sorption of diuron to soil is highly correlated with soil organic matter. In addition, diuron has -8 relatively low water solubility (42 ppm) and low volatility (6.9 x 10 mm Hg at 25° C). Although stable to hydrolysis at pH 5, 7, and 9, the minor degradate 3,4-dichloroaniline (3,4-DCA) was identified in all hydrolysis test solutions (0.5% of applied). The calculated half-lives for diuron in aqueous and soil photolysis studies were found to be 43 and 173 days respectively. In soil, the photolysis degradate were all minor, N'-(3,4-dichlorophenyl)-N-methylurea (DCPMU) at 3.6 percent, demethylated DCPMU (DCMU), 3,4-dichloroaniline (3,4- DCA), and 3,3',4,4'-tetrachlorobenzene (TCAB) at 0.7 percent, 0.370 ppm, and 0.038 15 ------- ppm respectively. The calculated half-lives in aerobic and anaerobic soil metabolism studies were 372 (aerobic) and 1,000 (anaerobic) days. Under aerobic conditions, the major degradate was DCPMU (20.9-22.5 % of the amount applied at 365 days), and minor degradates were DCPU and CO2. Under anaerobic conditions, the only major degradate identified was DCPMU, which accounted for a maximum of 10.3% of applied (at 45 days). In contrast to its persistence in laboratory studies of hydrolysis, aqueous and soil photolysis, and aerobic and anaerobic soil metabolism, diuron degraded relatively quickly in aquatic metabolism laboratory studies, with a half-life of 33 days under aerobic conditions and of 5 days under anaerobic conditions. The major metabolism degradate under aerobic conditions was N'-(3-chlorophenyl)-N,N-dimethylurea (MCPDMU) which reached 25 % of the applied dose by the end of the study and was evenly distributed between the soil and aqueous phase. Other degradates identified were DCPMU (9.2 percent) and CPMU (8 percent), and were primarily associated with the soil phase. The three major degradates under anaerobic conditions were MCPDMU which was mainly associated with the aqueous phase, l,l-dimethyl-3-phenylurea (PDMU), and CPMU. In terrestrial field dissipation studies in FL, MS, and CA with sand, silt loam, and silty clay loam soils, diuron dissipated in bare ground plots with half-lives of 73, 139, and 133 days, respectively. The major degradate DCPMU dissipated in the same plots with half-lives of 217, 1733, and 630 days. In aquatic field dissipation studies, half- lives were 115-177 days and the major degradate was DCPMU. These studies were conducted in drainage channel berms and soil, and thus should be compared quantitatively to soil laboratory studies, not aquatic studies. The environmental degradates of toxicological concern to humans and other non-target species include 3,4-DCA and TCAB. However, EPA's Metabolism Assessment Review Committee (MARC) has concluded that residues of 3,4-DCA is not a residue of concern for diuron as 3,4-DCA (<1%) was not a significant residue in any metabolism or hydrolysis study. Moreover, TCAB (0.038 ppm) residue was only found in the soil photolysis study which is a minor route of degradation for diuron. Degradation half-lives for the parent are presented in Table 2.1. This table also presents the major and minor degradates of the parent compound. The chemical structures of diuron and the metabolites (DCPMU, DCPU, DCA, and MCPDMU) are presented in Appendix B. 16 ------- Table 2.1 Summary of Diuron Half-lives Study Value (units) Major Dcgradatcs Minor Degradates IMRID # Hydrolysis Stable at pH 5, 7, 9 3,4-DCA (-2%) 41418804 Direct Aqueous Photolysis T ./2 = 43 days C02 41418805 Soil Photolysis T ./2 = 173 days DCPMU (3.6%) DCMU (< 0.7%), 3,4- DCA (0.370ppm), TCAB (0.038 ppm) 41719302 Aerobic Soil Metabolism T ./2 = 372 days DCPMU (22.5%) DCPU (3.4%), C02 (3.36%) 41719303 Anaerobic Soil Metabolism T ./2 = 1,000 days DCPMU (10.3 %) 41418806 Anaerobic Aquatic Metabolism T >/2 = 5 MCPDMU (83%) PDMU (13%), mCPMU (23%) 42661901 Aerobic Aquatic Metabolism T ¦/, = 33 MCPDMU (25%) DCPMU (9.2%), CPMU (8%) 42260501 Kd-ads / Kd-des (mL/g) 14 Material balances were not reported. 44490501 Terrestrial Field Dissipation Sand = 73 days Silt Loam =139 days Silty Clay Loam =133 days DCPMU (Sand = 217 days, Silt Loam = 1733 days, Silty Clay Loam = 630 days) 44654001 44865001 2.4.2 Environmental Transport Mechanisms AIR Diuron is applied by broadcast or band spray on soil surface using ground or aerial equipment, suggesting that there is a possibility of drift. Drift is quantitatively considered in the exposure assessment. However, diuron is non-volatile, as indicated by its low vapor pressure of 6.90 xlO"8 mm Hg (25° C), and a low Henry's law constant of 5.10 x 10"10 atm m3 mol"1. These properties indicate that diuron is unlikely to be dispersed in air over a large area and has a low tendency to volatilize from water or moist soils. Volatilization is insignificant except when diuron is exposed on the soil surface for several days or weeks under hot, dry conditions (Hess and Warren, 2002). 17 ------- WATER Diuron's K0c (468-1666) indicates a relatively low tendency to sorb to soils and sediments, while its hydrolysis and aqueous photolysis half-lives are relatively long. Consequently diuron is both mobile and relatively persistent, and is therefore prone to off-site movement in surface runoff, and migration to ground water. As discussed in the field monitoring studies below, diuron in runoff water is above Agency LOCs for direct effects on the CRLF (fish LC50 of 400 to 710 ppb). Andrieux et al. (1997) assessed losses of diuron via runoff from vineyards under natural Mediterranean climate conditions. During the growing season of 1994 and 1995, runoff and diuron concentrations were monitored at two field sites, one tilled and one under no- till management. Despite a time lag of greater than 4 months between application and the first runoff event in 1994, diuron concentrations in overland flow exceeded 200 ug/L at the no-till in both years of investigations. In 1995, the first strong rainfall following application removed 60% of total diuron runoff loss at both sites, although it accounted for 17 and 7% of the total seasonal runoff volume at the no-till and tilled site, respectively. In 1995, seasonal diuron loss at the no-till sites was 1.71 % and at the tilled site was 0.68%. No herbicide residues were detected in soil samples collected 1 week prior to chemical spraying in 1994 and 1995. The authors suggested that moderate temperatures and moisture conditions during winter facilitated complete microbial decomposition. In four-months of soil sampling following herbicide treatment there were no detections below a soil depth of 2 cm at the no-till plot. However, at the tilled site, pesticides were present in samples at depths of 0-15 cm 17 days after spraying to three months. The authors suggested that infiltration increases attributable to tillage effects accounted for diuron distribution in the soil profile. The authors concluded that the persistence of diuron coupled with tillage may provide the potential for leaching. In contrast, no-till practices have been shown to contribute to ground water contamination under certain California conditions. Braun and Hawkins (1991) conducted a rainfall runoff monitoring study in a citrus- growing region of Tulare County, California, where growers commonly avoid cultivation of citrus row middles. The row middles under such a management practice are typically compacted, with low infiltration rates and a high tendency to yield runoff during storm events. One method of disposing of excess runoff water in the area is to utilize dry wells that allow drainage of run off through the shallow hard-pan soil layer. This mechanism allows direct introduction of residues to the subsurface and subsequent potential migration to shallow ground water. Although diuron applications had occurred at least 2 months prior to sampling, relatively high concentrations of diuron in runoff water entering dry wells were found, ranging up to 890 |ig L"1. The data provide strong evidence that the widespread regional presence of diuron in ground water is at least partially attributable to contaminated runoff water entering dry wells. Spurlock et al. (1997) examined the runoff of diuron, simazine, and bromacil from citrus orchard middles under simulated rainfall conditions. Peak diuron runoff concentrations were comparable to those reported by Braun and Hawkins (1991), ranging from 600 to 18 ------- 1700 jug L"1. Little difference in runoff concentrations of the three pre-emergent herbicides was observed indicating that herbicide substitution would be an ineffective mitigation measure. They concluded that low soil permeability arising from hardpan layers or compaction drives the runoff process. These same factors are probably responsible for diuron off-site movement in runoff water from rights-of-way because rights-of-way are generally engineered to maximize water run off. Powell et al. (1996) studied off-site movement of diuron in surface water from a highway shoulder right-of-way application. Diuron was applied in a spray to a 2.4-meter wide strip next to the highway pavement, at a rate of 3.59 kg diuron ha"1. Simulated rain (13 mm in 1 hr) was applied to plots on treated highway shoulders at three sites. Diuron was detected at two of three sites. Concentrations of diuron were as high as 1770 |ig L"1 in runoff (combined water and sediment), from sampling conducted one day after herbicide application. From 0.2 to 5.4% of the diuron applied to highway shoulders moved off-site in runoff during the simulations. Simulated rain was applied to treated highway shoulder at intervals of 0, 2, and 4 wk after herbicide application. The highest percentages were observed when rain was simulated 1 day after herbicide application. Diuron is typically applied in winter or early spring during California's rainy season to control weeds. This practice, coupled with relatively high use, is one reason why diuron is commonly detected pesticides in California surface water. Diuron was detected in more than half of 955 surface water samples analyzed and reported in DPR's surface water database. Typical reporting limits for the diuron analyses are 0.05 -0.10 jug4. While detected concentrations range from 0.01 to 30.6 |ig L1, the majority of concentrations range from 0.1 to 1 |ig L'. These concentrations were approximately equal to the 20th and 80th percentiles of detected concentrations. The highest concentrations and most frequent detections occur during December-March, which coincides with both the rainy season and the peak diuron application season. Microbes are the primary agents in the degradation of diuron in aquatic environments. The aerobic biodegradation pathway for diuron is well established, proceeding by successive demethylation steps to form DCPMU, DCPU [l-(3,4-dichlorophenyl)urea] and DCA (3,4-dichloroaniline). Reductive dechlorination has been observed in anaerobic pond sediments and leads to the formation of the dechlorinated product, 3-(3- chlorophenyl)-l,l dimethylurea (Field et al., 1997). Boule et al. (1997) reported that the major photoproducts observed in the photolysis of diuron [3-(3,4-dichlorophenyl)-l,l- dimethylurea] in aqueous solution resulted from a heterolytic substitution of chlorine by OH. A wavelength effect was observed: at 254 nm the formation of 3-(4-chloro-3- hydroxyphenyl)-1,1-dimethylurea accounted for more than 90% of the conversion, whereas when the solution was irradiated in 'black light' (85% of photons emitted at 365 nm, about 7% at 344 nm), the major photoproduct was 3-(3-chloro-4-hydroxyphenyl)- 1,1-dimethylurea. However, as noted previously, photolysis is not generally a principal route of diuron degradation in aqueous systems. SOIL Diuron is moderately to highly persistent and mobile in soils. The commonly reported average field dissipation half-life is 115 days (MRID 44654001), although such half-lives 19 ------- are typically highly variable. Phytotoxic residues generally dissipate within a season when applied at low selective rates. At higher application rates, residues may persist for more than one year (Kidd and James, 1991). Microbial degradation is the primary means of diuron dissipation from soil. Photodegradation is not considered a primary dissipation route, but losses can be significant if diuron remains on the soil surface for several days or weeks (Hess and Warren, 2002). Mobility in the soil is related to organic matter and to the type of the residue. The metabolites are less mobile than the parent. Similar to many other pesticides, diuron sorption is highly correlated with organic matter (Spurlock andBiggar, 1994). Consequently leaching is greatest in low organic matter soils. Other soil conditions that favor diuron leaching include high soil permeability to water, such as in coarse soils. Due to the diuron's persistence and mobility, the herbicide is commonly detected in California's ground water (Troiano et al., 2001). As of June 2004, there were confirmed detections of diuron in 418 California water wells reported in DPR's well inventory database (WIDB). Typical diuron reporting limits in the WIDB are 0.05 |ig L"1, and detected diuron concentrations range up to 3.96 |ig L \ Diuron's use is regulated under DPR's ground water protection regulations (online at http://www.cdpr.ca.gov). In a right-of-way runoff study by Powell et al. (1996) post-simulation and end of season soil cores at the two sites with measurable runoff had detectable diuron residues only in the top 15 cm (0.5 ft). However, cores taken prior to application had diuron down to 0.30 m, presumably from the past year's application. At a third simulated rainfall site, no runoff left the plots. The absence of detectable residues in soil to a depth of 3.0 m (10 ft) suggests that much of the herbicide applied to the shoulder may have leached rapidly through the coarse gravelly soil. The soil data was insufficient to yield definite conclusions about leaching in infiltration areas. However, the levels of diuron found in runoff suggest that dry wells, if present, would provide an important conduit for the transport of diuron to groundwater. The authors concluded that rights-of-ways with high percentages of gravel and sand in the soil may be of concern, since diuron may be transported rapidly to depths greater than 0.3 m. Bogarets et al. (2000) studied diuron's microbial degradation and ecotoxicology to investigate its fate after application to soils. Quantitative biodegradation assays were executed with fungal strains, showing that diuron was degraded but not entirely mineralized. A series of tests were carried out to choose the most efficient fungal strain for diuron degradation. Among the fungal strains tested, only three were able to transform diuron to any extent (up to 50%) after 7 days of incubation: B. bassiana, C. elegans, and M. isabellina. No degradation occurred using fungal strains: F. oxysporum and G. candidum. Although C. elegans was the most efficient since there was no diuron remaining after seven days of incubation. Diuron degradation by the three fungal strains led to the formation of two metabolites obtained in different ratios according to the microorganism. For the three fungal strains, diuron degradation led to the formation of the demethylated products. The identified metabolites were synthesized in sufficient amounts to confirm their structures and determine their non-target toxicity using four biotests. According to the Microtox test, the metabolites N- (3,4-dichlorophenyl)-N- 20 ------- methylurea and N-3, 4-dichlorophenylurea presented a three times higher toxicity than that of diuron. 2.4.3 Mechanism of Action Diuron is a systemic substituted phenylurea herbicide. Diuron is taken up by plants and rapidly translocated. Diuron primarily functions by inhibiting the Hill reaction in photosynthesis, limiting the production of high-energy compounds such as adenosine triphosphate (ATP) used for various metabolic processes. Diuron binds to the Qb- binding niche on D1 protein of the photosystem II complex in chloroplast thylakoid membranes, thus blocking electron transport from QAto Qb. This process prevents CO2 fixation and the production of ATP and other high energy compounds which are needed for plant growth. The inability to reoxidize Qa promotes the formation of triplet state chlorophyll, which interacts with ground state oxygen to form singlet oxygen. Both triplet chlorophyll and singlet oxygen can extract hydrogen from unsaturated lipids, producing a lipid radical and initiating a chain reaction of lipid peroxidation. Lipids and proteins are attacked and oxidized, resulting in loss of chlorophyll and carotenoids, and in leaky membranes which cause cells and cell organelles to dry and disintegrate rapidly (Hess and Warren, 2002). 2.4.4 Use Characterization There are currently 72 product registrations for diuron including two Special Local Needs registrations in California. Diuron contains two significant impurities from the manufacturing process 3,3',4,4'- tetrachloroazobenzene (TCAB) and 3,3',4,4'-tetrachloroazoxybenzene (TCAOB), both 'dioxin-like' substances that are known to cause a skin disease known as chloracne. TCAB levels between 0.15 and 28 ppm have been found in diuron samples tested. TCAOB is present at lower levels. Since these residues appear to be present at insignificant levels, they are not assessed further in this assessment. In addition to agricultural uses, diuron also has widespread use in non-agricultural applications, especially industrial and rights of way uses, where often in combination with other herbicides it provides total vegetation control. These applications include along fence lines, pipelines, powerlines, railway lines, roads, footpaths; in timber yards and storage areas; and around commercial, industrial and farm buildings, electrical substations, and petroleum storage tanks. It has some use as an algaecide in ornamental ponds, fountains, and aquaria, but not natural water bodies. Diuron is one of the most commonly used pesticides in California. Reported statewide use in 2002 was 1,303,745 pounds. Of this reported use, 48 percent was applied to rights of way, followed by 18 percent and 14 percent on alfalfa and oranges, respectively. Additional 2002 uses in California included grapes (4.3 percent), landscape maintenance (3.7 percent), walnuts (2.6 percent), and cotton (1.6 percent). Diuron use has been reported in all 58 counties in California. 21 ------- Diuron is often used in combination with other herbicides such as bromacil, hexazinone, paraquat, thiadiazuron, imazapyr monosodium, sodium chlorate, sodium metaborate, and copper sulfate (U.S. EPA, 2004a). Diuron is available in wettable powder, granular, flowable, pelleted/tableted, liquid suspension, and soluble concentrate formulations. However, true granular applications are not allowed in California and are therefore not assessed further in this document. Diuron is applied using the following equipment: groundboom sprayer, aerial equipment, chemigation, rights-of-way sprayer, high-pressure handwand, low-pressure handwand, tractor-drawn spreader, granular backpack spreader (since true granular applications are not allowed in California, this application method will not be assessed), push-type spreader, airless sprayer, paintbrush, shaker-type applicator, backpack sprayer, belly grinder, and by hand. . For agricultural uses, labeled single application rates range from 0.4 to 12 lbs active ingredient (ai) per acre (A). For non-agricultural uses labeled rates are 12 lbs ai/acre. Diuron may be applied to non-agricultural areas 1 to 2 times per year. A national map showing the estimated poundage of diuron used for agricultural purposes in 2002 by county is presented in Figure 2.1. The map was obtained from the U. S. Geological Survey (USGS), National Water Quality Assessment Program (NAWQA) website (URL: http://water.usgs.gov/nawqa/pnsp/usage/maps/show_map.php?year=02&map=ml991). 22 ------- DIURON - herbicide 2002 estimated annual agricultural use Crops Total Percent pounds applied national use citrus fruit 1233611 35.75 cotton 1001843 29.03 sugarcane 396903 11.50 alfalfa hay 290978 8.43 grapes 127492 3.69 apples 69420 2.01 asparagus 66251 1.92 field and grass seed crop 61094 1.77 pecans 42615 1.23 walnuts 34338 1.00 Average annual use of active ingredient (pounds per square mile of agricultural land in county) I I no estimated use ~ 0.001 to 0.006 ~ 0.007 to 0.03 ~ 0.031 to 0.133 ~ 0.134 to 1.026 ¦ >=1.027 Source: USGS, 2002 Figure 2.1 Diuron Agricultural Use in Total Pounds per County Analysis of labeled use information is the critical first step in evaluating the federal action. The current label for diuron represents the FIFRA regulatory action, therefore, labeled use and application rates specified on the label form the basis of this assessment. The assessment of use information is critical to the development of the action area and selection of appropriate modeling scenarios and inputs. Table 2.2 presents the uses allowed in California that will be focused on in this assessment along with the corresponding scenarios used for the Tier II aquatic modeling. Please see Section 3.0 for further explanation. 23 ------- Table 2.2 Active Use Sites for Diuron as Applicable to Ca ifornia ('nip Si'i'ii;iriii AGRICULTURAL RIGHTS-OF-WAY/FENCEROWS/HEDGEROWS CA impervious RLF and CA residential RLF ALFALFA CA alfalfa OP APPLE, PEACH, PEAR CA fruit STD ARTICHOKE, ASPARGUS CA row crop RLF BANANA, PLANTAIN, PAPAYA CA avocado RLF BLACKBERRY, BOYSENBERRY, DEWBERRY, LOGANBERRY, RASPBERRY (BLACK/RED) OR Berries OP BLUEBERRY CA wine grapes RLF CITRUS CA citrus STD CORN, FIELD CA corn OP FILBERT (HAZELNUT), PECAN, WALNUT (ENGLISH/BLACK) CA almond STD GRAPE CA grapes STD OLIVE CAOliveRLF - V2 PEPPERMINT, SPEARMINT OR mint STD SORGHUM, WHEAT CA wheat RLF \.iii-I-'.hi(I/I-\v(I ( nips COTTON CA cotton STD Nuii-Cnip I.iiihI AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS. PAVED AREAS (PRIVATE ROADS/SIDEWALKS) CA impervious RLF BERMUDAGRASS CA rangeland hay RLF GRASSES GROWN FOR SEED CA turf RLF IRRIGATION SYSTEMS CA residential RLF ORNAMENTAL HERBACEOUS PLANTS CA Nursery NON-AGRICULTURAL RIGHTS OF WAY CA impervious RLF and CA residential RLF UNCULTIVATED AG, UNCULTIVATED NON-AG CA Rangeland Hay_V2 RLF * Uses provided by BEAD - July 23, 2008, Scenarios provided by EFED 24 ------- The Agency's Biological and Economic Analysis Division (BEAD) provides an analysis of both national- and county-level usage information (Kaul and Jones, 2006) using state- level usage data obtained from USDA-NASS1, Doane (www.doane.com); the full dataset is not provided due to its proprietary nature) and the California's Department of Pesticide Regulation Pesticide Use Reporting (CDPR PUR) database2 . CDPR PUR is considered a more comprehensive source of usage data than USDA-NASS or EPA proprietary databases, and thus the usage data reported for diuron by county in this California- specific assessment were generated using CDPR PUR data. Four years (2002-2005) of usage data were included in this analysis. Available data from CDPR PUR were obtained for every pesticide application made on every use site at the section level (approximately one square mile) of the public land survey system. BEAD summarized these data to the county level by site, pesticide, and unit treated. Calculating county-level usage involved summarizing across all applications made within a section and then across all sections within a county for each use site and for each pesticide. The county level usage data that were calculated include: average annual pounds applied, average annual area treated, and average and maximum application rate across all five years. The units of area treated are also provided where available. A summary of diuron usage for all California use sites is provided in Appendix C, and a summary of the CDPR PUR usage data can be found in Appendix D. 2.5 Assessed Species The CRLF was federally listed as a threatened species by USFWS effective June 24, 1996 (USFWS 1996). It is one of two subspecies of the red-legged frog and is the largest native frog in the western United States (USFWS 2002). A brief summary of information regarding CRLF distribution, reproduction, diet, and habitat requirements is provided in Sections 2.5.1 through 2.5.4, respectively. Further information on the status, distribution, and life history of and specific threats to the CRLF is provided in Attachment I. Final critical habitat for the CRLF was designated by USFWS on April 13, 2006 (USFWS 2006; 71 FR 19244-19346). Further information on designated critical habitat for the CRLF is provided in Section 2.6. 2.5.1 Distribution The CRLF is endemic to California and Baja California (Mexico) and historically inhabited 46 counties in California including the Central Valley and both coastal and interior mountain ranges (USFWS 1996). Its range has been reduced by about 70%, and the species currently resides in 22 counties in California (USFWS 1996). The species has an elevational range of near sea level to 1,500 meters (5,200 feet) (Jennings and Hayes 1 United States Depart of Agriculture (USDA), National Agricultural Statistics Service (NASS) Chemical Use Reports provide summary pesticide usage statistics for select agricultural use sites by chemical, crop and state. See http://www.usda.eov/nass/pubs/estindxl,htm#agchem. 2 The California Department of Pesticide Regulation's Pesticide Use Reporting database provides a census of pesticide applications in the state. See http://www.cdpr.ca.gov/docs/pur/purmain.htm. 25 ------- 1994); however, nearly all of the known CRLF populations have been documented below 1,050 meters (3,500 feet) (USFWS 2002). Populations currently exist along the northern California coast, northern Transverse Ranges (USFWS 2002), foothills of the Sierra Nevada (5-6 populations), and in southern California south of Santa Barbara (two populations) (Fellers 2005a). Relatively larger numbers of CRLFs are located between Marin and Santa Barbara Counties (Jennings and Hayes 1994). A total of 243 streams or drainages are believed to be currently occupied by the species, with the greatest numbers in Monterey, San Luis Obispo, and Santa Barbara counties (USFWS 1996). Occupied drainages or watersheds include all bodies of water that support CRLFs (i.e., streams, creeks, tributaries, associated natural and artificial ponds, and adjacent drainages), and habitats through which CRLFs can move (i.e., riparian vegetation, uplands) (USFWS 2002). The distribution of CRLFs within California is addressed in this assessment using four categories of location including recovery units, core areas, designated critical habitat, and known occurrences of the CRLF reported in the California Natural Diversity Database (CNDDB) that are not included within core areas and/or designated critical habitat (see Figure 2.2). Recovery units, core areas, and other known occurrences of the CRLF from the CNDDB are described in further detail in Attachment I, and designated critical habitat is addressed in Section 2.6. Recovery units are large areas defined at the watershed level that have similar conservation needs and management strategies. The recovery unit is primarily an administrative designation, and land area within the recovery unit boundary is not exclusively CRLF habitat. Core areas are smaller areas within the recovery units that comprise portions of the species' historic and current range and have been determined by USFWS to be important in the preservation of the species. Designated critical habitat is generally contained within the core areas, although a number of critical habitat units are outside the boundaries of core areas, but within the boundaries of the recovery units. Additional information on CRLF occurrences from the CNDDB is used to cover the current range of the species not included in core areas and/or designated critical habitat, but within the recovery units. Other Known Occurrences from the CNDBB The CNDDB provides location and natural history information on species found in California. The CNDDB serves as a repository for historical and current species location sightings. Information regarding known occurrences of CRLFs outside of the currently occupied core areas and designated critical habitat is considered in defining the current range of the CRLF. See: http://www.dfg.ca.gov/bdb/html/cnddb info.html for additional information on the CNDDB. 26 ------- Recovery Units 1. Sierra Nevada Foothills and Central Valley 2. North Coast Range Foothills and Western Sacramento River Valley 3. North Coast and North San Francisco Bay 4. South and East San Francisco Bay 5. Central Coast 6. Diablo Range and Salinas Valley 7. Northern Transverse Ranges and Tehachapi Mountains 8. Southern Transverse and Peninsular Ranges Legend CD Recovery Unit Boundaries ^ |j Currently Occupied Core Areas | Critical Habitat | CNDDB Occurence Sections County Boundaries g 180 Miles _l Core Areas 1. Feather River 19. Watsonville Slough-Elkhorn Slough 2. Yuba River- S. Fork Feather River 20. Carmel River — Santa Lucia 3. Traverse Creek Middle Fork/ American R. Rubicon 21. Gahlan Range 4. Cosumnes River 22. Estero Bay 5. South Fork Calaveras River* 23. Arroyo Grange River 6. Tuolumne River* 24. Santa Maria River — Santa Ynez River 7. Piney Creek* 25. Sisquoc River 8. Cottonwood Creek 26. Ventura River — Santa Clara River 9. Putah Creek - Cache Creek* 27. Santa Monica Bay —Venura Coastal Streams 10. Lake Berryessa Tributaries 28. Estrella River 11. Upper Sonoma Creek 29. San Gabriel Mountain* 12. Petaluma Creek — Sonoma Creek 30. Forks of the Mojave* 13. Pt. Reyes Peninsula 31. Santa Ana Mountain* 14. Belvedere Lagoon 32. Santa Rosa Plateau 15. Jameson Canyon - Lower Napa River 33. San Luis Ray* 16. East San Francisco Bay 34. Sweetwater* 17. Santa Clara Valley 35. Laguna Mountain* 18. South San Francisco Bay * Core areas that were historically occupied by the California red-legged frog are not included in the map figure 2.2 Recovery Unit, Core Area, Critical Habitat, and Occurrence Designations for CRLF 27 ------- Other Known Occurrences from the CNDBB The CNDDB provides location and natural history information on species found in California. The CNDDB serves as a repository for historical and current species location sightings. Information regarding known occurrences of CRLFs outside of the currently occupied core areas and designated critical habitat is considered in defining the current range of the CRLF. See: http://www.dfg.ca.gov/bdb/html/cnddb info.html for additional information on the CNDDB. 2.5.2 Reproduction CRLFs breed primarily in ponds; however, they may also breed in quiescent streams, marshes, and lagoons (Fellers 2005a). According to the Recovery Plan (USFWS 2002), CRLFs breed from November through late April. Peaks in spawning activity vary geographically; Fellers (2005b) reports peak spawning as early as January in parts of coastal central California. Eggs are fertilized as they are being laid. Egg masses are typically attached to emergent vegetation, such as bulrushes (Scirpus spp.) and cattails (Typha spp.) or roots and twigs, and float on or near the surface of the water (Hayes and Miyamoto 1984). Egg masses contain approximately 2000 to 6000 eggs ranging in size between 2 and 2.8 mm (Jennings and Hayes 1994). Embryos hatch 10 to 14 days after fertilization (Fellers 2005a) depending on water temperature. Egg predation is reported to be infrequent and most mortality is associated with the larval stage (particularly through predation by fish); however, predation on eggs by newts has also been reported (Rathburn 1998). Tadpoles require 11 to 28 weeks to metamorphose into juveniles (terrestrial-phase), typically between May and September (Jennings and Hayes 1994, USFWS 2002); tadpoles have been observed to over-winter (delay metamorphosis until the following year) (Fellers 2005b, USFWS 2002). Males reach sexual maturity at 2 years, and females reach sexual maturity at 3 years of age; adults have been reported to live 8 to 10 years (USFWS 2002). Figure 2.3 depicts CRLF annual reproductive timing. J F M A M J J A S o N D Light Blue = Breeding/Egg Masses Green = Tadpoles (except those that over-winter) Orange = Adults and juveniles can be present all year figure 2.3 - CRLF Reproductive Events by Month 2.5.3 Diet Although the diet of CRLF aquatic-phase larvae (tadpoles) has not been studied specifically, it is assumed that their diet is similar to that of other frog species, with the aquatic phase feeding exclusively in water and consuming diatoms, algae, and detritus (USFWS 2002). Tadpoles filter and entrap suspended algae (Seale and Beckvar, 1980) 28 ------- via mouthparts designed for effective grazing of periphyton (Wassersug, 1984, Kupferberg et al.\ 1994; Kupferberg, 1997; Altig and McDiarmid, 1999). Juvenile and adult CRLFs forage in aquatic and terrestrial habitats, and their diet differs greatly from that of larvae. The main food source for juvenile aquatic- and terrestrial- phase CRLFs is thought to be aquatic and terrestrial invertebrates found along the shoreline and on the water surface. Hayes and Tennant (1985) report, based on a study examining the gut content of 35 juvenile and adult CRLFs, that the species feeds on as many as 42 different invertebrate taxa, including Arachnida, Amphipoda, Isopoda, Insecta, and Mollusca. The most commonly observed prey species were larval alderflies (Sialis cf. californica), pillbugs (Armadilliadrium vulgare), and water striders (Gerris sp). The preferred prey species, however, was the sowbug (Hayes and Tennant, 1985). This study suggests that CRLFs forage primarily above water, although the authors note other data reporting that adults also feed under water, are cannibalistic, and consume fish. For larger CRLFs, over 50% of the prey mass may consists of vertebrates such as mice, frogs, and fish, although aquatic and terrestrial invertebrates were the most numerous food items (Hayes and Tennant 1985). For adults, feeding activity takes place primarily at night; for juveniles feeding occurs during the day and at night (Hayes and Tennant 1985). 2.5.4 Habitat CRLFs require aquatic habitat for breeding, but also use other habitat types including riparian and upland areas throughout their life cycle. CRLF use of their environment varies; they may complete their entire life cycle in a particular habitat or they may utilize multiple habitat types. Overall, populations are most likely to exist where multiple breeding areas are embedded within varying habitats used for dispersal (USFWS 2002). Generally, CRLFs utilize habitat with perennial or near-perennial water (Jennings et al. 1997). Dense vegetation close to water, shading, and water of moderate depth are habitat features that appear especially important for CRLF (Hayes and Jennings 1988). Breeding sites include streams, deep pools, backwaters within streams and creeks, ponds, marshes, sag ponds (land depressions between fault zones that have filled with water), dune ponds, and lagoons. Breeding adults have been found near deep (0.7 m) still or slow moving water surrounded by dense vegetation (USFWS 2002); however, the largest number of tadpoles have been found in shallower pools (0.26 - 0.5 m) (Reis, 1999). Data indicate that CRLFs do not frequently inhabit vernal pools, as conditions in these habitats generally are not suitable (Hayes and Jennings 1988). CRLFs also frequently breed in artificial impoundments such as stock ponds, although additional research is needed to identify habitat requirements within artificial ponds (USFWS 2002). Adult CRLFs use dense, shrubby or emergent vegetation closely associated with deep-water pools bordered with cattails and dense stands of overhanging vegetation (http://www.fws.gov/endangered/features/rl frog/rlfrog.html#where). In general, dispersal and habitat use depends on climatic conditions, habitat suitability, and life stage. Adults rely on riparian vegetation for resting, feeding, and dispersal. The foraging quality of the riparian habitat depends on moisture, composition of the plant 29 ------- community, and presence of pools and backwater aquatic areas for breeding. CRLFs can be found living within streams at distances up to 3 km (2 miles) from their breeding site and have been found up to 30 m (100 feet) from water in dense riparian vegetation for up to 77 days (USFWS 2002). During dry periods, the CRLF is rarely found far from water, although it will sometimes disperse from its breeding habitat to forage and seek other suitable habitat under downed trees or logs, industrial debris, and agricultural features (UWFWS 2002). According to Jennings and Hayes (1994), CRLFs also use small mammal burrows and moist leaf litter as habitat. In addition, CRLFs may also use large cracks in the bottom of dried ponds as refugia; these cracks may provide moisture for individuals avoiding predation and solar exposure (Alvarez 2000). 2.6 Designated Critical Habitat In a final rule published on April 13, 2006, 34 separate units of critical habitat were designated for the CRLF by USFWS (USFWS 2006; FR 51 19244-19346). A summary of the 34 critical habitat units relative to USFWS-designated recovery units and core areas (previously discussed in Section 2.5.1) is provided in Table 2.4. 'Critical habitat' is defined in the ESA as the geographic area occupied by the species at the time of the listing where the physical and biological features necessary for the conservation of the species exist, and there is a need for special management to protect the listed species. It may also include areas outside the occupied area at the time of listing if such areas are 'essential to the conservation of the species.' All designated critical habitat for the CRLF was occupied at the time of listing. Critical habitat receives protection under Section 7 of the ESA through prohibition against destruction or adverse modification with regard to actions carried out, funded, or authorized by a federal Agency. Section 7 requires consultation on federal actions that are likely to result in the destruction or adverse modification of critical habitat. To be included in a critical habitat designation, the habitat must be 'essential to the conservation of the species.' Critical habitat designations identify, to the extent known using the best scientific and commercial data available, habitat areas that provide essential life cycle needs of the species or areas that contain certain primary constituent elements (PCEs) (as defined in 50 CFR 414.12(b)). PCEs include, but are not limited to, space for individual and population growth and for normal behavior; food, water, air, light, minerals, or other nutritional or physiological requirements; cover or shelter; sites for breeding, reproduction, rearing (or development) of offspring; and habitats that are protected from disturbance or are representative of the historic geographical and ecological distributions of a species. The designated critical habitat areas for the CRLF are considered to have the following PCEs that justify critical habitat designation: • Breeding aquatic habitat; • Non-breeding aquatic habitat; • Upland habitat; and 30 ------- • Dispersal habitat. Further description of these habitat types is provided in Attachment 1. Occupied habitat may be included in the critical habitat only if essential features within the habitat may require special management or protection. Therefore, USFWS does not include areas where existing management is sufficient to conserve the species. Critical habitat is designated outside the geographic area presently occupied by the species only when a designation limited to its present range would be inadequate to ensure the conservation of the species. For the CRLF, all designated critical habitat units contain all four of the PCEs, and were occupied by the CRLF at the time of FR listing notice in April 2006. The FR notice designating critical habitat for the CRLF includes a special rule exempting routine ranching activities associated with livestock ranching from incidental take prohibitions. The purpose of this exemption is to promote the conservation of rangelands, which could be beneficial to the CRLF, and to reduce the rate of conversion to other land uses that are incompatible with CRLF conservation. Please see Attachment 1 for a full explanation on this special rule. USFWS has established adverse modification standards for designated critical habitat (USFWS 2006). Activities that may destroy or adversely modify critical habitat are those that alter the PCEs and jeopardize the continued existence of the species. Evaluation of actions related to use of diuron that may alter the PCEs of the CRLF's critical habitat form the basis of the critical habitat impact analysis. According to USFWS (2006), activities that may affect critical habitat and therefore result in adverse effects to the CRLF include, but are not limited to the following: (1) Significant alteration of water chemistry or temperature to levels beyond the tolerances of the CRLF that result in direct or cumulative adverse effects to individuals and their life-cycles. (2) Significant increase in sediment deposition within the stream channel or pond or disturbance of upland foraging and dispersal habitat that could result in elimination or reduction of habitat necessary for the growth and reproduction of the CRLF by increasing the sediment deposition to levels that would adversely affect their ability to complete their life cycles. (3) Significant alteration of channel/pond morphology or geometry that may lead to changes to the hydrologic functioning of the stream or pond and alter the timing, duration, water flows, and levels that would degrade or eliminate the CRLF and/or its habitat. Such an effect could also lead to increased sedimentation and degradation in water quality to levels that are beyond the CRLF's tolerances. (4) Elimination of upland foraging and/or aestivating habitat or dispersal habitat. (5) Introduction, spread, or augmentation of non-native aquatic species in stream segments or ponds used by the CRLF. (6) Alteration or elimination of the CRLF's food sources or prey base (also evaluated as indirect effects to the CRLF). 31 ------- As previously noted in Section 2.1, the Agency believes that the analysis of direct and indirect effects to listed species provides the basis for an analysis of potential effects on the designated critical habitat. Because diuron is expected to directly impact living organisms within the action area, critical habitat analysis for diuron is limited in a practical sense to those PCEs of critical habitat that are biological or that can be reasonably linked to biologically mediated processes. 2.7 Action Area For listed species assessment purposes, the action area is considered to be the area affected directly or indirectly by the federal action and not merely the immediate area involved in the action (50 CFR 402.02). It is recognized that the overall action area for the national registration of diuron is likely to encompass considerable portions of the United States based on the large array of agricultural uses. However, the scope of this assessment limits consideration of the overall action area to those portions that may be applicable to the protection of the CRLF and its designated critical habitat within the state of California. The Agency's approach to defining the action area under the provisions of the Overview Document (USEPA 2004) considers the results of the risk assessment process to establish boundaries for that action area with the understanding that exposures below the Agency's defined LOCs constitute a no-effect threshold. For the purposes of this assessment, attention will be focused on the footprint of the action (i.e., the area where pesticide application occurs), plus all areas where offsite transport (i.e., spray drift, downstream dilution, etc.) may result in potential exposure within the state of California that exceeds the Agency's LOCs. Deriving the geographical extent of this portion of the action area is based on consideration of the types of effects that diuron may be expected to have on the environment, the exposure levels to diuron that are associated with those effects, and the best available information concerning the use of diuron and its fate and transport within the state of California. Specific measures of ecological effect for the CRLF that define the action area include any direct and indirect toxic effect to the CRLF and any potential modification of its critical habitat, including reduction in survival, growth, and fecundity as well as the full suite of sublethal effects available in the effects literature. Therefore, the action area extends to a point where environmental exposures are below any measured lethal or sublethal effect threshold for any biological entity at the whole organism, organ, tissue, and cellular level of organization. In situations where it is not possible to determine the threshold for an observed effect, the action area is not spatially limited and is assumed to be the entire state of California. The definition of action area requires a stepwise approach that begins with an understanding of the federal action. The federal action is defined by the currently labeled uses for diuron. An analysis of labeled uses and review of available product labels was completed. Several of the currently labeled uses are special local needs (SLN) uses or are restricted to specific states and are excluded from this assessment. In addition, a distinction has been made between food use crops and those that are non-food/non- agricultural uses. For those uses relevant to the CRLF, the analysis indicates that, for 32 ------- diuron, agricultural and non-agricultural uses listed in Table 2.2 are considered part of the federal action evaluated in this assessment. Following a determination of the assessed uses, an evaluation of the potential "footprint" of diuron use patterns (i.e., the area where pesticide application occurs) is determined. This "footprint" represents the initial area of concern, based on an analysis of available land cover data for the state of California. The initial area of concern is defined as all land cover types and the stream reaches within the land cover areas that represent the labeled uses described above. A map representing all the land cover types related to agricultural uses that make up the initial area of concern for diuron is presented in Figure 2.4. Diuron, in fact, is used in all 58 of California's counties when you consider non- agricultural uses as well. 33 ------- Diruon use &CRLF overlap CNDDB occurrence sections Critical habitat Core areas County boundaries Diuron Use & CRLF Habitat Produced 12/2/2008 ¦tb=mh=ihb Kil o mete rs 0 20 40 80 120 160 Compiled from California Count)' boundaries (ESRI, 2002), USCW GapAnalysis Program Orchard/ Vineyard Landcover (GAP) National Land Cewer Database (NLCD) (MRLC, 2001) Map created by US Environmental Protection Agency, Office of Pesticides Programs, Environmental Fate and Effects Division. Projection: Albers Equal Area Conic USGS, North American Datum of 1983 (NAD 1983). Figure 2.4 Initial area of concern, or "footprint" of potential use, for diuron 34 ------- Once the initial area of concern is defined, the next step is to define the potential boundaries of the action area by determining the extent of offsite transport via spray drift and runoff where exposure of one or more taxonomic groups to the pesticide exceeds the listed species LOCs. As previously discussed, the action area is defined by the most sensitive measure of direct and indirect ecological toxic effects including reduction in survival, growth, reproduction, and the entire suite of sublethal effects from valid, peer-reviewed studies. As stated in the 2003 Registration Eligibility Decision (RED), diuron has been characterized as a "known/likely" human carcinogen, based on urinary bladder carcinomas in both sexes of the Wistar rat, kidney carcinomas in the male rat (a rare tumor), and mammary gland carcinomas in the female NMRI mouse. Because of this, the spatial extent of the action area (i.e., the boundary where exposures and potential effects are less than the Agency's LOC) for diuron cannot be determined. Therefore, it is assumed that the action area encompasses the entire state of California, regardless of the spatial extent (i.e., initial area of concern or footprint) of the pesticide use(s). This determination is supported by the fact that diuron use has been reported in all 58 counties in California. The AgDRIFT model (Version 2.01) is used to define how far from the initial area of concern an effect to a given species may be expected via spray drift. The spray drift analysis for diuron using the most sensitive endpoint (most sensitive plant toxicity study: Vegetative Vigor - Tomato (Lycopersicon esculentum)) suggests that a maximum spray drift distance of at least 1,000 feet is necessary. The Tier 1 ground analysis allows users to evaluate off-site deposition and exposure out to 1,000 ft downwind from the location of the application. Further detail on the spray drift analysis is provided in Section 3.2.5. In addition to the buffered area from the spray drift analysis, the final action area also considers the downstream extent of diuron that exceeds the LOC (discussed in Section 3.2.6). An evaluation of usage information was conducted to determine the area where use of diuron may impact the CRLF. This analysis is used to characterize where predicted exposures are most likely to occur, but does not preclude use in other portions of the action area. A more detailed review of the county-level use information was also completed. These data suggest that diuron has historically been used on a wide variety of agricultural and non-agricultural uses. 2.8 Assessment Endpoints and Measures of Ecological Effect Assessment endpoints are defined as "explicit expressions of the actual environmental value that is to be protected."3 Selection of the assessment endpoints is based on valued entities (e.g., CRLF, organisms important in the life cycle of the CRLF, and the PCEs of its designated critical habitat), the ecosystems potentially at risk (e.g., waterbodies, riparian vegetation, and upland and dispersal habitats), the migration pathways of diuron 3 From U.S. EPA (1992). Framework for Ecological Risk Assessment. EPA/630/R-92/001. 35 ------- (e.g., runoff, spray drift, etc.), and the routes by which ecological receptors are exposed to diuron (e.g., direct contact, etc.). 2.8.1. Assessment Endpoints for the CRLF Assessment endpoints for the CRLF include direct toxic effects on the survival, reproduction, and growth of the CRLF, as well as indirect effects, such as reduction of the prey base or modification of its habitat. In addition, potential modification of critical habitat is assessed by evaluating potential effects to PCEs, which are components of the habitat areas that provide essential life cycle needs of the CRLF. Each assessment endpoint requires one or more "measures of ecological effect," defined as changes in the attributes of an assessment endpoint or changes in a surrogate entity or attribute in response to exposure to a pesticide. Specific measures of ecological effect are generally evaluated based on acute and chronic toxicity information from registrant-submitted guideline tests that are performed on a limited number of organisms. Additional ecological effects data from the open literature are also considered. It should be noted that assessment endpoints are limited to direct and indirect effects associated with survival, growth, and fecundity, and do not include the full suite of sublethal effects used to define the action area. According the Overview Document (USEPA 2004), the Agency relies on acute and chronic effects endpoints that are either direct measures of impairment of survival, growth, or fecundity or endpoints for which there is a scientifically robust, peer reviewed relationship that can quantify the impact of the measured effect endpoint on the assessment endpoints of survival, growth, and fecundity. A complete discussion of all the toxicity data available for this risk assessment, including resulting measures of ecological effect selected for each taxonomic group of concern, is included in Section 4 of this document. A summary of the assessment endpoints and measures of ecological effect selected to characterize potential assessed direct and indirect CRLF risks associated with exposure to diuron is provided in Table 2.4. Table 2.4 Assessment Endpoints and Measures of Ecological Effects Assessment Ijulpoinl Measures of l-eolouieal I-Heels' Aquatic-Phase CRLF (Eggs, larvae, juveniles, and adults"f Direct Effects 1. Survival, growth, and reproduction of CRLF la. Amphibian acute LC50 (ECOTOX) or most sensitive fish acute LC50 (guideline or ECOTOX) if no suitable amphibian data are available lb. Amphibian chronic NOAEC (ECOTOX) or most sensitive fish chronic NOAEC (guideline or ECOTOX) lc. Amphibian early-life stage data (ECOTOX) or most sensitive fish early-life stage NOAEC (guideline or ECOTOX) Indirect Effects and Critical Habitat Effects 2. Survival, growth, and reproduction of CRLF individuals via indirect effects on 2a. Most sensitive fish, aquatic invertebrate, and aquatic plant EC50 or LC50 (guideline or ECOTOX) 4 All registrant-submitted and open literature toxicity data reviewed for this assessment are included in Appendix J. 36 ------- Assessment l-ndpoinl Measures of l-colouical I-fleets' aquatic prey food supply (i.e., fish, freshwater invertebrates, non-vascular plants) 2b. Most sensitive aquatic invertebrate and fish chronic NOAEC (guideline or ECOTOX) 3. Survival, growth, and reproduction of CRLF individuals via indirect effects on habitat, cover, food supply, and/or primary productivity (i.e., aquatic plant community) 3 a. Vascular plant acute EC50 (duckweed guideline test or ECOTOX vascular plant) 3b. Non-vascular plant acute EC50 (freshwater algae or diatom, or ECOTOX non-vascular) 4. Survival, growth, and reproduction of CRLF individuals via effects to riparian vegetation 4a. Distribution of EC25 values for monocots (seedling emergence, vegetative vigor, or ECOTOX) 4b. Distribution of EC25 values for dicots (seedling emergence, vegetative vigor, or ECOTOX) Terrestrial-Phase CRLF (Juveniles and adults) Direct Effects 5. Survival, growth, and reproduction of CRLF individuals via direct effects on terrestrial phase adults and juveniles 5a. Most sensitive birdb or terrestrial-phase amphibian acute LC50 or LD50 (guideline or ECOTOX) 5b. Most sensitive birdb or terrestrial-phase amphibian chronic NOAEC (guideline or ECOTOX) Indirect Effects and Critical Habitat Effects 6. Survival, growth, and reproduction of CRLF individuals via effects on terrestrial prey (i.e., terrestrial invertebrates, small mammals, and frogs) 6a. Most sensitive terrestrial invertebrate and vertebrate acute EC50 or LC50 (guideline or ECOTOX)0 6b. Most sensitive terrestrial invertebrate and vertebrate chronic NOAEC (guideline or ECOTOX) 7. Survival, growth, and reproduction of CRLF individuals via indirect effects on habitat (i.e., riparian and upland vegetation) 7a. Distribution of EC25 for monocots (seedling emergence, vegetative vigor, or ECOTOX 7b. Distribution of EC25 for dicots (seedling emergence, vegetative vigor, or ECOTOX) a Adult frogs are no longer in the "aquatic phase" of the amphibian life cycle; however, submerged adult frogs are considered "aquatic" for the purposes of this assessment because exposure pathways in the water are considerably different that exposure pathways on land. b Birds are used as surrogates for terrestrial phase amphibians. 2.8.2 Assessment Endpoints for Designated Critical Habitat As previously discussed, designated critical habitat is assessed to evaluate actions related to the use of diuron that may alter the PCEs of the CRLF's critical habitat. PCEs for the CRLF were previously described in Section 2.6. Actions that may modify critical habitat are those that alter the PCEs and jeopardize the continued existence of the CRLF. Therefore, these actions are identified as assessment endpoints. It should be noted that evaluation of PCEs as assessment endpoints is limited to those of a biological nature (i.e., the biological resource requirements for the listed species associated with the critical habitat) and those for which diuron effects data are available. Adverse modification to the critical habitat of the CRLF includes, but is not limited to, the following, as specified by USFWS (2006): 1. Alteration of water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs. 37 ------- 2. Alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs. 3. Significant increase in sediment deposition within the stream channel or pond or disturbance of upland foraging and dispersal habitat. 4. Significant alteration of channel/pond morphology or geometry. 5. Elimination of upland foraging and/or aestivating habitat, as well as dispersal habitat. 6. Introduction, spread, or augmentation of non-native aquatic species in stream segments or ponds used by the CRLF. 7. Alteration or elimination of the CRLF's food sources or prey base. Measures of such possible effects by labeled use of diuron on critical habitat of the CRLF are described in Table. Some components of these PCEs are associated with physical abiotic features (e.g., presence and/or depth of a water body, or distance between two sites), which are not expected to be measurably altered by use of pesticides. Assessment endpoints used for the analysis of designated critical habitat are based on the adverse modification standard established by USFWS (2006). 38 ------- Table 2.5 Summary of Assessment Endpoints and Measures of Ecological Effect for Primary Constituent Elements of Designated Critical Habitat" Assessment Endpoint Measures of Ecological Effect Aquatic-Phase CRLF PCEs (Aquatic Breeding Habitat and Aquatic Non-Breeding Habitat) Alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond: aquatic habitat (including riparian vegetation) provides for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. a. Most sensitive aquatic plant EC50 (guideline or ECOTOX) b. Distribution of EC25 values for terrestrial monocots (seedling emergence, vegetative vigor, or ECOTOX) c. Distribution of EC25 values for terrestrial dicots (seedling emergence, vegetative vigor, or ECOTOX) Alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food source. a. Most sensitive EC50 values for aquatic plants (guideline or ECOTOX) b. Distribution of EC25 values for terrestrial monocots (seedling emergence or vegetative vigor, or ECOTOX) c. Distribution of EC25 values for terrestrial dicots (seedling emergence, vegetative vigor, or ECOTOX) Alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source. a. Most sensitive EC50 or LC50 values for fish or aquatic- phase amphibians and aquatic invertebrates (guideline or ECOTOX) b. Most sensitive NOAEC values for fish or aquatic-phase amphibians and aquatic invertebrates (guideline or ECOTOX) Reduction and/or modification of aquatic-based food sources for pre-metamorphs (e.g., algae) a. Most sensitive aquatic plant EC50 (guideline or ECOTOX) Terrestrial-Phase CRLF PCEs (Upland Habitat and Dispersal Habitat) Elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs: Upland areas within 200 ft of the edge of the riparian vegetation or dripline surrounding aquatic and riparian habitat that are comprised of grasslands, woodlands, and/or wetland/riparian plant species that provides the CRLF shelter, forage, and predator avoidance a. Distribution of EC25 values for monocots (seedling emergence, vegetative vigor, or ECOTOX) b. Distribution of EC25 values for dicots (seedling emergence, vegetative vigor, or ECOTOX) c. Most sensitive food source acute EC50/LC50 and NOAEC values for terrestrial vertebrates (mammals) and invertebrates, birds or terrestrial-phase amphibians, and freshwater fish. Elimination and/or disturbance of dispersal habitat: Upland or riparian dispersal habitat within designated units and between occupied locations within 0.7 mi of each other that allow for movement between sites including both natural and altered sites which do not contain barriers to dispersal Reduction and/or modification of food sources for terrestrial phase juveniles and adults Alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. a Physico-chemical water quality parameters such as salinity, pH, and hardness are not evaluated because these processes are not biologically mediated and, therefore, are not relevant to the endpoints included in this assessment. 39 ------- 2.9 Conceptual Model 2.9.1 Risk Hypotheses Risk hypotheses are specific assumptions about potential adverse effects {i.e., changes in assessment endpoints) and may be based on theory and logic, empirical data, mathematical models, or probability models (U.S. EPA, 1998). For this assessment, the risk is stressor-linked, where the stressor is the release of diuron to the environment. The following risk hypotheses are presumed for this endangered species assessment: The labeled use of diuron within the action area may: • Directly affect the CRLF by causing mortality or by adversely affecting growth or fecundity; • Indirectly affect the CRLF by reducing or changing the composition of food supply; • Indirectly affect the CRLF or modify designated critical habitat by reducing or changing the composition of the aquatic plant community in the ponds and streams comprising the species' current range and designated critical habitat, thus affecting primary productivity and/or cover; • Indirectly affect the CRLF or modify designated critical habitat by reducing or changing the composition of the terrestrial plant community (i.e., riparian habitat) required to maintain acceptable water quality and habitat in the ponds and streams comprising the species' current range and designated critical habitat; • Modify the designated critical habitat of the CRLF by reducing or changing breeding and non-breeding aquatic habitat (via modification of water quality parameters, habitat morphology, and/or sedimentation); • Modify the designated critical habitat of the CRLF by reducing the food supply required for normal growth and viability of juvenile and adult CRLFs; • Modify the designated critical habitat of the CRLF by reducing or changing upland habitat within 200 ft of the edge of the riparian vegetation necessary for shelter, foraging, and predator avoidance. • Modify the designated critical habitat of the CRLF by reducing or changing dispersal habitat within designated units and between occupied locations within 0.7 mi of each other that allow for movement between sites including both natural and altered sites which do not contain barriers to dispersal. • Modify the designated critical habitat of the CRLF by altering chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs. 2.9.2 Diagram The conceptual model is a graphic representation of the structure of the risk assessment. It specifies the diuron release mechanisms, biological receptor types, and effects endpoints of potential concern. The conceptual models for aquatic and terrestrial phases of the CRLF are shown in Figure 2.5 and Figure 2.6, respectively, and the conceptual models for the aquatic and terrestrial PCE components of critical habitat are shown in 40 ------- Figure 2.7 and Figure 2.8, respectively. Exposure routes shown in dashed lines are not quantitatively considered because the contribution of those potential exposure routes to potential risks to the CRLF and modification to designated critical habitat is expected to be negligible. Figure 2.5 Conceptual Model for Aquatic-Phase of the CRLF 41 ------- Figure 2.6 Conceptual Model for Terrestrial-Phase of the CRLF 42 ------- Figure 2.7 Conceptual Model for Pesticide Effects on Aquatic Component of CRLF Critical Habitat 43 ------- Figure 2.8 Conceptual Model for Pesticide Effects on Terrestrial Component of CRLF Critical Habitat 2.10 Analysis Plan In order to address the risk hypothesis, the potential for direct and indirect effects to the CRLF, its prey, and its habitat is estimated. In the following sections, the use, environmental fate, and ecological effects of diuron are characterized and integrated to assess the risks. This is accomplished using a risk quotient (ratio of exposure concentration to effects concentration) approach. Although risk is often defined as the likelihood and magnitude of adverse ecological effects, the risk quotient-based approach does not provide a quantitative estimate of likelihood and/or magnitude of an adverse effect. However, as outlined in the Overview Document (U.S. EPA, 2004), the likelihood of effects to individual organisms from particular uses of diuron is estimated using the probit dose-response slope and either the level of concern (discussed below) or actual calculated risk quotient value. 2.10.1 Measures to Evaluate the Risk Hypothesis and Conceptual Model 2.10.1.1 Measures of Exposure The environmental fate properties of diuron along with available monitoring data indicate that runoff and spray drift are the principle potential transport mechanisms of diuron to the aquatic and terrestrial habitats of the CRLF. In this assessment, transport of diuron 44 ------- through runoff and spray drift is considered in deriving quantitative estimates of diuron exposure to CRLF, its prey and its habitats. The reported vapor pressure of diuron is 6.9 x 10"8 torr at 25°C; therefore, volatilization is not considered a probable route of dissipation. Measures of exposure are based on aquatic and terrestrial models that predict estimated environmental concentrations (EECs) of diuron using maximum labeled application rates and methods of application. The models used to predict aquatic EECs are the Pesticide Root Zone Model coupled with the Exposure Analysis Model System (PRZM/EXAMS). T-REX is used to predict terrestrial EECs on food items. TerrPlant is used to derive EECs relevant to terrestrial and wetland plants. These models are parameterized using relevant reviewed registrant-submitted environmental fate data. PRZM (v3.12.2, May 2005) and EXAMS (v2.98.4.6, April 2005) are screening simulation models coupled with the input shell pe5.pl (Aug 2007) to generate daily exposures and l-in-10 year EECs of diuron that may occur in surface water bodies adjacent to application sites receiving diuron through runoff and spray drift. PRZM simulates pesticide application, movement and transformation on an agricultural field and the resultant pesticide loadings to a receiving water body via runoff, erosion and spray drift. EXAMS simulates the fate of the pesticide and resulting concentrations in the water body. The standard scenario used for ecological pesticide assessments assumes application to a 10-hectare agricultural field that drains into an adjacent 1-hectare water body, 2-meters deep (20,000 m3 volume) with no outlet. PRZM/EXAMS was used to estimate screening-level exposure of aquatic organisms to diuron. The measure of exposure for aquatic species is the l-in-10 year return peak or rolling mean concentration. The l-in-10 year peak is used for estimating acute exposures of direct effects to the CRLF, as well as indirect effects to the CRLF through effects to potential prey items, including: algae, aquatic invertebrates, fish and frogs. The l-in-10-year 60-day mean is used for assessing chronic exposure to the CRLF and fish and frogs serving as prey items; the 1-in-10-year 21-day mean is used for assessing chronic exposure for aquatic invertebrates, which are also potential prey items. Exposure estimates for the terrestrial-phase CRLF and terrestrial invertebrates and mammals (serving as potential prey) assumed to be in the target area or in an area exposed to spray drift are derived using the T-REX model (version 1.3.1, 12/07/2006). This model incorporates the Kenega nomograph, as modified by Fletcher et al. (1994), which is based on a large set of actual field residue data. The upper limit values from the nomograph represented the 95th percentile of residue values from actual field measurements (Hoerger and Kenega, 1972). For modeling purposes, direct exposures of the CRLF to diuron through contaminated food are estimated using the EECs for the small bird (20 g) which consumes small insects. Dietary-based and dose-based exposures of potential prey (small mammals) are assessed using the small mammal (15 g) which consumes short grass. The small bird (20g) consuming small insects and the small mammal (15g) consuming short grass are used because these categories represent the largest RQs of the size and dietary categories in T-REX that are appropriate surrogates 45 ------- for the CRLF and one of its prey items. Estimated exposures of terrestrial insects to diuron are bound by using the dietary based EECs for small insects and large insects. Birds are currently used as surrogates for terrestrial-phase CRLF. However, amphibians are poikilotherms (body temperature varies with environmental temperature) while birds are homeotherms (temperature is regulated, constant, and largely independent of environmental temperatures). Therefore, amphibians tend to have much lower metabolic rates and lower caloric intake requirements than birds or mammals. As a consequence, birds are likely to consume more food than amphibians on a daily dietary intake basis, assuming similar caloric content of the food items. Therefore, the use of avian food intake allometric equation as a surrogate to amphibians is likely to result in an over- estimation of exposure and risk for reptiles and terrestrial-phase amphibians. Therefore, T-REX (version 1.3.1) has been refined to the T-HERPS model (v. 1.0), which allows for an estimation of food intake for poikilotherms using the same basic procedure as T-REX to estimate avian food intake. EECs for terrestrial plants inhabiting dry and wetland areas are derived using TerrPlant (version 1.2.2, 12/26/2006). This model uses estimates of pesticides in runoff and in spray drift to calculate EECs. EECs are based upon solubility, application rate and minimum incorporation depth. Spray drift model, AgDRIFT is used to assess exposures of terrestrial phase CRLF and its prey to diuron deposited on terrestrial habitats by spray drift. In addition to the buffered area from the spray drift analysis, the downstream extent of diuron that exceeds the LOC for the effects determination is also considered. 2.10.1.2 Measures of Effect Data identified in Section 2.8 are used as measures of effect for direct and indirect effects to the CRLF. Data were obtained from registrant submitted studies or from literature studies identified by ECOTOX. The ECOTOXicology database (ECOTOX) was searched in order to provide more ecological effects data and in an attempt to bridge existing data gaps. ECOTOX is a source for locating single chemical toxicity data for aquatic life, terrestrial plants, and wildlife. ECOTOX was created and is maintained by the USEPA, Office of Research and Development, and the National Health and Environmental Effects Research Laboratory's Mid-Continent Ecology Division. The assessment of risk for direct effects to the terrestrial-phase CRLF makes the assumption that toxicity of diuron to birds is similar to or less than the toxicity to the terrestrial-phase CRLF. The same assumption is made for fish and aquatic-phase CRLF. Algae, aquatic invertebrates, fish, and amphibians represent potential prey of the CRLF in the aquatic habitat. Terrestrial invertebrates, small mammals, and terrestrial-phase amphibians represent potential prey of the CRLF in the terrestrial habitat. Aquatic, semi- aquatic, and terrestrial plants represent habitat of CRLF. 46 ------- The acute measures of effect used for animals in this screening level assessment are the LD50, LC50 and EC50. LD stands for "Lethal Dose", and LD50 is the amount of a material, given all at once, that is estimated to cause the death of 50% of the test organisms. LC stands for "Lethal Concentration" and LC50 is the concentration of a chemical that is estimated to kill 50% of the test organisms. EC stands for "Effective Concentration" and the EC50 is the concentration of a chemical that is estimated to produce a specific effect in 50% of the test organisms. Endpoints for chronic measures of exposure for listed and non-listed animals are the NOAEL/NOAEC and NOEC. NOAEL stands for "No Ob served-Adverse-Effect-Level" and refers to the highest tested dose of a substance that has been reported to have no harmful (adverse) effects on test organisms. The NOAEC (i.e., "No-Observed-Adverse-Effect-Concentration") is the highest test concentration at which none of the observed effects were statistically different from the control. The NOEC is the No-Observed-Effects-Concentration. For non-listed plants, only acute exposures are assessed (i.e., EC25 for terrestrial plants and EC50 for aquatic plants). It is important to note that the measures of effect for direct and indirect effects to the CRLF and its designated critical habitat are associated with impacts to survival, growth, and fecundity, and do not include the full suite of sublethal effects used to define the action area. According the Overview Document (USEPA 2004), the Agency relies on effects endpoints that are either direct measures of impairment of survival, growth, or fecundity or endpoints for which there is a scientifically robust, peer reviewed relationship that can quantify the impact of the measured effect endpoint on the assessment endpoints of survival, growth, and fecundity. 2.10.1.3 Integration of Exposure and Effects Risk characterization is the integration of exposure and ecological effects characterization to determine the potential ecological risk from agricultural and non-agricultural uses of diuron, and the likelihood of direct and indirect effects to CRLF in aquatic and terrestrial habitats. The exposure and toxicity effects data are integrated in order to evaluate the risks of adverse ecological effects on non-target species. For the assessment of diuron risks, the risk quotient (RQ) method is used to compare exposure and measured toxicity values. EECs are divided by acute and chronic toxicity values. The resulting RQs are then compared to the Agency's levels of concern (LOCs) (USEPA, 2004) (see Appendix E). For this endangered species assessment, listed species LOCs are used for comparing RQ values for acute and chronic exposures of diuron directly to the CRLF. If estimated exposures directly to the CRLF of diuron resulting from a particular use are sufficient to exceed the listed species LOC, then the effects determination for that use is "may affect". When considering indirect effects to the CRLF due to effects to animal prey (aquatic and terrestrial invertebrates, fish, frogs, and mice), the listed species LOCs are also used. If estimated exposures to CRLF prey of diuron resulting from a particular use are sufficient to exceed the listed species LOC, then the effects determination for that use is a "may affect." If the RQ being considered also exceeds the non-listed species acute risk LOC, then the effects determination is a LAA. If the acute RQ is between the listed species 47 ------- LOC and the non-listed acute risk species LOC, then further lines of evidence {i.e. probability of individual effects, species sensitivity distributions) are considered in distinguishing between a determination of NLAA and a LAA. When considering indirect effects to the CRLF due to effects to algae as dietary items or plants as habitat, the non- listed species LOC for plants is used because the CRLF does not have an obligate relationship with any particular aquatic and/or terrestrial plant. If the RQ being considered for a particular use exceeds the non-listed species LOC for plants, the effects determination is "may affect". Further information on LOCs is provided in Appendix E. 2.10.2 Data Limitations No acceptable toxicity studies have submitted to the Agency, nor were any acceptable studies found in the open literature for the chronic effects of diuron on avian or amphibian species. Linuron, a similar chemical of the same class, has acceptable chronic avian data. The avian reproductive studies for linuron (MRID 42541801, 42541802) using mallard duck and bobwhite quail found reproductive effects at 300 ppm ai with an NOAEL being established at 100 ppm. The endpoints affected for mallard duck reproduction are egg production, adult body weight, feed consumption, viable embryos of eggs set, number of viable embryos, and number of live embryos. For the Bobwhite Quail the endpoints affected are hatchability and offspring survivability. The LC50 for the Bobwhite Quail exposed to linuron was 1700ppm, where the LC50 for the Bobwhite Quail exposed to linuron was almost identical at 1730 ppm (MRID 00022923). Therefore it is assumed that linuron and diuron have a similar mode of action and therefore chronic effects from diuron are assumed to be similar to those observed from linuron exposure. Chronic risks are therefore assumed to amphibian species and their avian surrogates for uses of diuron in California. 3.0 Exposure Assessment Diuron is a broad-spectrum residual herbicide registered for pre-emergent and post- emergent control of both broadleaf and annual grassy weeds. In the U.S. diuron is used on a variety of fruit and nut crops, grains, cotton, corn, sorghum, mint, gum, asparagus, sugarcane, seed crops, coffee, hay, cut flowers, and for fallow and idle cropland use. It may be used in irrigation and drainage systems when water is not present. It is also used to control weeds on hard surfaces, such as, roads, railway tracks, and paths (at around 3 kg/ha), and to control weeds in crops, such as, pear and apple trees, forestry, ornamental trees and shrubs, pineapples, sugar cane, cotton, alfalfa and wheat (at lower rates of around 1.8 kg/ha). Its use in some locations is becoming limited due to the development of resistant weed species. In some products it is formulated with other active ingredients such as glyphosate, bromacil, hexazinone, amitrole and 2,4-D. This assessment evaluates only those registered uses which allow use in California. 48 ------- 3.1 Label Application Rates and Intervals Currently registered agricultural and non-agricultural uses of diuron within California include agricultural and non-agricultural crops, as well as industrial and paved areas. The uses and applicable PRZM/EXAMS scenarios being assessed were summarized in Table 2.2. For a list of the application rates and intervals used, please refer to Table 3.2 below. 3.2 Aquatic Exposure Assessment 3.2.1 Modeling Approach As discussed in Section 2.10.1.1, the models used to predict aquatic EECs are the Pesticide Root Zone Model coupled with the Exposure Analysis Model System (PRZM/EXAMS). Using PRZM/EXAMS, aquatic exposures are quantitatively estimated for all of the assessed uses with scenarios that represent high exposure sites for diuron use. Each of these sites represents a 10 hectare field that drains into a 1-hectare pond that is 2 meters deep and has no outlet. Exposure estimates generated using the standard pond are intended to represent a wide variety of vulnerable water bodies that occur at the top of watersheds including prairie pot holes, playa lakes, wetlands, vernal pools, man-made and natural ponds, and intermittent and first-order streams. As a group, there are factors that make these water bodies more or less vulnerable than the standard surrogate pond. Static water bodies that have larger ratios of drainage area to water body volume would be expected to have higher peak EECs than the standard pond. These water bodies will be either shallower or have large drainage areas (or both). Shallow water bodies tend to have limited additional storage capacity, and thus, tend to overflow and carry pesticide in the discharge whereas the standard pond has no discharge. As watershed size increases beyond 10 hectares, at some point, it becomes unlikely that the entire watershed is planted to a single crop, which is all treated with the pesticide. Headwater streams can also have peak concentrations higher than the standard pond, but they tend to persist for only short periods of time and are then carried downstream. Crop-specific management practices for all of the assessed uses of diuron were used for modeling, including application rates, number of applications per year, application intervals, and the first application date for each crop. The date of first application was developed based on several sources of information including data provided by BEAD, a summary of individual applications from the CDPR PUR data, and Crop Profiles maintained by the USD A. 3.2.2 Model Inputs The input parameters for PRZM and EXAMS are presented in Table 3.1. 49 ------- Table 3.1 Summary of PRZIVl/EXAMS Environmental Fate Data Used for Aquatic Exposure Inputs for Diuron Endangered Species Assessment for the CRLF Fate Property Value (unit) MRID (or source) Molecular Weight 233.1 g/mol Product Chemistry Henry's constant 5. IE-10 atm-m"3/mol MRID 00017763 MRID 43762901, Herbicide Vapor Pressure 6.9E-8 torr (25°C) Handbook 7th Edition, WSSA, 1994 Solubility in Water 420 mg/1 (42 mg/1 x 10 per input guidance) Product Chemistry, Input Parameter Guidance Photolysis in Water 43 days MRID 41418805 Aerobic Soil Metabolism Half-lives 372 days MRID 41719303 Hydrolysis Stable at pH7 (>500 hrs) MRID 41418804 Aerobic Aquatic Metabolism (water column) 99 days (33 days x 3 per input guidance) MRID 42260501, Input Parameter Guidance Anaerobic Aquatic Metabolism (benthic) 15 days (5 days x 3 per input guidance) MRID 42661901, Input Parameter Guidance Koc 920 (avg. of 468, 626, and 1666) MRID 44490501 Application rate and frequency Various (see Table 3.2) Per Label Instructions Application intervals Various (see Table 3.2) Per Label Instructions Chemical Application Method (CAM) 1 Label; Input Guidance for Eco Assessments 0.99 Ground 0.95 Aerial Per Input Guidance for Eco Application Efficiency Assessments Spray Drift Fraction1 0.01 Ground 0.05 Aerial Per Input Guidance for Eco Assessments Inputs determined in accordance with EFED "Guidance for Chemistry and Management Practice Input Parameters for Use in Modeling the Environmental Fate and Transport of Pesticides" dated February 28, 2002 3.2.3 Results The PRZM/EXAMS model estimated aquatic EECs for the various scenarios and application practices are listed in Table 3.2. See Appendix G for a summary of the outputs. Peak EECs ranged from 3.76 to 4911 ppb for use on grasses grown for seed and paved areas (impervious surfaces), respectively. Since some of the application data needed for modeling were not stated on the labels, assumptions were made by EFED analysts regarding the maximum number of applications allowed per season, and/or the interval between applications. The assumptions were as follows: 50 ------- • For the application intervals that were not stated, the most conservative (minimum) known application interval was used (21 days for cotton). 21 days was chosen because it was the minimum known interval that was registered. • If the maximum number of applications was also missing, the maximum number of intervals a year was determine by taking the maximum allowable application of active ingredient per year (12 lb ai/yr) and dividing it by the maximum application rate for the use in question. The following is an example of a calculation to determine the maximum number of applications per year for the use of diuron on blackberries: Max No. of Appl. Yr"1 = 12 lb ai. acre _1 yr _1 ^ 2.4 lb ai. acre _1 appl._1 = 5 appl. yr"1 Table 3.2 Aquatic EECs (ppb) for Diuron Uses in California ( l'M|l V|i|iliiiiliiiii Kiilc ill)- Vi Mux ' mI V|i|ili(iiliMii<. 1 ill t-r\:il |{rl\\ rill V|>|><. V pplii-iil i«»n iiiHIhmI IVsik i:i:c |)|)h •)(, II ¦ I.I. ( plili 21 1 >n \ i:i:< phi) (.11 l)il\ I.I. ( pllll 1 \|)|)^ XI I I l)il\M AGRICULTURAL RIGHTS-OF- WAY/FENCEROWS/HEDGEROWS 12 2 21 Ground 688 672 584 457 ALFALFA 2.4 1 0 Ground 10.6 10.4 9.1 7.2 Aircraft 13.7 13.3 11.7 9.4 APPLE 3.2 2 90 Ground 11.7 11.2 9.6 7.8 ARTICHOKE 3.2 1 0 Ground 32.0 31.1 27.4 25.1 ASPARGUS 3.2 1 0 Ground 37.0 35.5 30.2 22.9 BANANA, PLANTAIN 4.8 2 42 Ground 54.2 52.0 44.3 32.8 BLACKBERRY, BOYSENBERRY 2.4 5 21 Ground 140 136 121 103 BLUEBERRY 1.6 2 112 Ground 4.65 4.48 4.05 3.21 CITRUS 3.2 2 60 Ground 6.83 6.57 5.65 4.88 CORN, FIELD 0.8 1 0 Ground 7.24 6.95 6.13 4.68 DEWBERRY 2.4 3 60 Ground 19.7 19.3 16.8 14.3 FILBERT (HAZELNUT) 2.2 2 150 Ground 18.9 18.4 15.9 12.2 GRAPE 3.2 2 90 Ground 9.71 9.32 8.10 6.37 LOGANBERRY, RASPBERRY (BLACK/RED) 2.4 1 0 Ground 5.13 4.95 4.31 3.02 OLIVE 1.6 2 168 Ground 7.17 6.90 5.97 4.86 PAPAYA 4 1 0 Ground 25.2 24.6 21.2 15.7 PEPPERMINT 2.4 1 0 Ground 4.17 4.05 3.55 2.79 PEACH 3 1 0 Ground 11.0 10.5 9.0 7.3 PEAR 3.2 1 0 Ground 11.7 11.2 9.6 7.8 PECAN 3.2 1 0 Ground 27.0 26.3 22.8 17.3 SORGHUM 0.4 2 21 Ground 5.87 5.67 5.44 4.52 SPEARMINT 2.4 5 21 Ground 107 104 97.1 82.0 WALNUT (ENGLISH/BLACK) 3 2 150 Ground 25.8 25.1 21.8 16.6 WHEAT 1.6 1 0 Ground 18.0 17.3 14.9 12.0 12.8 12.3 11.9 10.3 Null I'lHIll I'CCll ( |M|>- l 1 >| |t >\ 1.6 3 21 Ground 8.36 8.13 7.42 5.72 Aircraft 14.2 13.9 12.9 10.2 Null ( iiip 1 mill AIRPORTS LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS 12 2 21 Ground 2190 2115 1825 1382 BERMUDAGRASS 2.4 5 21 Ground 226 219 204 168 GRASSES GROWN FOR SEED 1.6 1 0 Ground 3.76 3.62 3.20 2.89 51 ------- M:i\ mI' VppliciliMll- |||1(T\ ill Urlw till A|>|>» Vpplifiiliiin IIIClllMll IRRIGATION SYSTEMS 12 2 21 Ground 52.6 50.7 45.0 37.5 ORNAMENTAL HERBACEOUS PLANTS 4 1 0 Ground 36.5 35.5 30.7 23.6 NON-AGRICULTURAL RIGHTS OF WAY 12 2 90 Ground 630 619 560 476 Aircraft 627 616 552 468 PAVED AREAS (PRIVATE ROADS/SIDEWALKS) 12 1 0 Ground 4911 4725 4241 3428 UNCULTIVATED AG 12 1 0 Ground 1130 1099 1023 840 Aircraft 1169 1130 1063 891 UNCULTIVATED NON-AG 12 2 21 Ground 285 276 252 203 3.2.4 Existing Monitoring Data 3.2.4.1 USGS NAWQA Surface Water Data A critical step in the process of characterizing EECs is comparing the modeled estimates with available surface water monitoring data. The USGS has collected 347 surface water samples from four river basins in California (Sacramento, San Joaquin-Tulare, Santa Ana, and Nevada) with a detection limit of 0.01 ppb. Out of these samples, 232 (69%) were positive for diuron. The maximum concentration observed was 23.3 ppb in the Santa Ana River Basin in San Bernardino County. 3.2.4.2 USGS NAWQA Groundwater Data The USGS collected 428 ground water samples where 75 (17.5%) of these samples contained diuron. The three river basins looked at were Sacramento, San Joaquin-Tulare, and Santa Ana. The maximum concentration observed was 1.8 ppb in the San Joaquin- Tulare water basins in Kings County. Similar to the most recent analysis of USGS data, an "Analysis of Diuron Monitoring Data with [an] emphasis on the USGS NAQWA Surface Water Sampling" in February 1999, observed the San Joaquin-Tulare basin as well. It was concluded that diuron concentrations reached their peaks in the winter and spring seasons (January to June). USGS concluded that runoff was more efficient than irrigation return flows in transporting pesticides, in this case diuron, into surface water. Although USGS detected diuron in more than 10% of the ground water samples, the concentrations were generally less than 0.1 ppb. 3.2.4.3 California Department of Pesticide Regulation Data The California Department of Pesticide Regulation (CDPR) has been collecting surface water data on diuron since 1992. In a memo sent to the Special Review and Reregi strati on Division (SRRD) from the Environmental Fate and Effects Division (EFED) in 2003, titled "Surface Water Monitoring Data for Diuron", summarized surface water monitoring data for diuron collected by CDPR over the period December 2000 to March 2001. There were three locations (82 samples) in the Sacramento River, and two locations (54 samples) in the San Joaquin River. One-hundred percent of the samples 52 ------- taken from the San Joaquin River had detectable diuron, with a maximum concentration of 8.45 ppb. The average concentration at the San Joaquin River stations was 1.7 ppb. About seventy-five percent of the samples in the Sacramento River had detectable diuron with a maximum concentration of 1.42 ppb. The average concentration, assuming that all non-detects were equal to the detection limit of 0.05 ppb, was 0.16 ppb. Currently, the monitoring data available at CDPR has expanded to include data collected from 2004, 2005, and 2006. Between 2004 and 2006, the data indicates that diuron concentrations range from non-detect to 160 ppb (San Joaquin County, January 3, 2005, and Tulare County, February 21, 2005). Out of 156 sampling locations, 152 locations (97.4%) were located within 0.5 miles or less of an agricultural area. The four other locations were 8 miles or more from the nearest agricultural area. Sixteen percent of the 705 samples collected from 2004 to 2006 were greater than the detection limit. The majority of the higher concentrations of diuron came from San Joaquin County, and occurred between the end of December and the beginning of January. The year 2005, for San Joaquin County, yielded the greatest concentrations overall with an average concentration of 10.9 ppb for the year. Diuron's concentrations being greatest in, and mainly only showing up in the monitoring data during the winter months is consistent with the labeled use for pre-emergence application. In addition, during the winter months, high concentrations may be detected due to transport from rainfall runoff. Runoff during the winter in the San Joaquin Valley may include large amounts of non-agricultural runoff from coastal ranges. When comparing this monitoring data to the results from PRZM/EXAMS, the CDPR data further supports that spray drift is a major route of off site movement. 3.2.4.4 Atmospheric Monitoring Data The California Air Review Board has not conducted ambient air monitoring of diuron. Therefore, no atmospheric monitoring data is available. 3.2.6 Downstream Dilution Analysis To complete this assessment, the greatest ratio of aquatic RQ to LOC was estimated. Using an assumption of uniform runoff across the landscape, it is assumed that streams flowing through treated areas (i.e. the initial area of concern) are represented by the modeled EECs; as those waters move downstream, it is assumed that the influx of non- impacted water will dilute the concentrations of diuron present. Using the LC50 for Selenastrum capricornutum (the most sensitive species) of 2.40 ppb and a maximum peak EEC for applications to non-agricultural rights-of- way/fencerows/hedgerows of 341.6 ug/L yields an RQ/LOC ratio of 142.33. Using the downstream dilution approach (described in more detail in Appendix F) yields a target percent crop area (PCA). This value has been input into the downstream dilution approach resulting in a total of 285 kilometers of stream downstream from the initial area 53 ------- of concern (footprint of use). Similar to the spray drift buffer described above, the LAA/NLAA determination is based on the area defined by the point where concentrations exceed the LC50 value, in this case 2.40 ppb. See Appendix F for the spatial summary of diuron used to determine the downstream dilution. 3.3 Terrestrial Animal Exposure Assessment T-REX (Version 1.3.1) is used to calculate dietary and dose-based EECs of diuron for the CRLF and its potential prey (e.g. small mammals and terrestrial insects) inhabiting terrestrial areas. EECs used to represent the CRLF are also used to represent exposure values for frogs serving as potential prey of CRLF adults. T-REX simulates a 1-year time period. For this assessment, ground spray applications of diuron are considered, as discussed in below. Terrestrial EECs for ground spray applications of diuron were derived for the uses registered in California. Given that no data on interception and subsequent dissipation from foliar surfaces is available for diuron, a default foliar dissipation half-life of 35 days is used based on the work of Willis and McDowell (1987). Use specific input values, including number of applications, and application rate are shown in Table 3.3. An example output from T-REX is available in Appendix H. Table 3.3 T-REX Model Input Parameters Crop Application Kale per Crop Scenario (Ihs/A) Nil in her of Applications Application interval (davs) Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non- Agricultural Areas 12 2 21 Alfalfa, Peppermint 2.4 1 n/a Apple, Grape 3.2 2 90 Citrus 3.2 2 60 Artichoke, Asparagus 3.2 1 n/a Banana, Plantain 4.8 2 42 Bermuda grass, Blackberry, Boysenberry, Spearmint 2.4 5 21 Blueberry 1.6 2 112 Field Corn 0.8 1 n/a Cotton 1.6 3 21 Dewberry 2.4 3 60 Filbert (Hazelnut) 2.2 2 150 Grass Seed, Wheat 1.6 1 n/a 54 ------- Crop Application Kale per Crop Scenario (lbs/A) .Number of Applications Application interval (days) Loganberry, Raspberry 2.4 l n/a Olive 1.6 2 168 Pecan, Pear 3.2 1 n/a Ornamental Herbaceous Plants, Papaya 4 1 21 Uncultivated Agricultural Areas, Paved Areas 12 1 n/a Peach 3 1 n/a Sorghum 0.4 1 n/a Walnut 3 2 150 T-REX is also used to calculate EECs for terrestrial insects exposed to diuron. Dietary- based EECs calculated by T-REX for small and large insects (units of a.i. /g) are used to bound an estimate of exposure to bees. Available acute contact toxicity data for bees exposed to diuron (in units of |ig a.i. /bee), are converted to |ig a.i. /g (of bee) by multiplying by 1 bee/0.128 g. The EECs are later compared to the adjusted acute contact toxicity data for bees in order to derive RQs. For modeling purposes, exposures of the CRLF to diuron through contaminated food are estimated using the EECs for the small bird (20 g) which consumes small insects. Dietary-based and dose-based exposures of potential prey are assessed using the small mammal (15 g) which consumes short grass. Upper-bound Kenega nomogram values reported by T-REX for these two organism types are used for derivation of EECs for the CRLF and its potential prey (Table 3.4). Dietary-based EECs for small and large insects reported by T-REX as well as the resulting adjusted EECs are available in Table 3.5. Table 3.4 Upper-bound Kenega Nomogram EECs for Dietary- and Dose-based Exposures of the CRLF and its Prey to Diuron i:i.( s for CKI.I KIX s for Prey (small mammals) I sc Dictarv- Dose-based Dictarv- Dosc-bascd bascd i:i:c (ppin) i:i:c (mg/kg-bw) bascd i:i:c (ppm) 1 -1A (mg/kg- bw) Agricultural Rights-of- Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non- 2688.80 3062.28 4780.09 5444.05 55 ------- i:i.( s lor ( KI.I KIX s for Prcv (small mammals) I sc Dictarv- Dosc-hascd Dictarv- Dosc-hascd r i.v • bascd i:i:( (ppm) i: i :c (m«/k«-h\v) hascd i:ix (ppm) 1-1A (m«/kg- b\v) Agricultural Areas Alfalfa, Peppermint, 324.00 369.00 576.00 549.17 Apple, Grape 504.68 574.78 897.21 855.42 Artichoke, Asparagus 432.00 874.68 768.00 732.23 Banana, Plantain 930.06 1059.24 1653.44 1576.43 Bermuda grass, Blackberry, Boysenberry, 833.22 948.95 1481.28 1412.29 Spearmint Blueberry 239.50 272.77 425.79 405.95 Citrus, 563.65 641.94 1002.05 955.38 Field Corn 108.00 123.00 192.00 183.06 Cotton 452.53 515.38 804.49 767.02 Dewberry 452.83 515.73 805.03 767.54 Filbert (Hazelnut) 312.23 355.60 555.07 529.22 Grass Seed, Wheat 216.00 246.00 384.00 366.11 Loganberry, Raspberry 324.00 369.00 576.00 549.17 Olive 223.75 254.83 397.78 379.26 Ornamental Herbaceous plants, Papaya 540.00 615.01 960.00 915.29 Uncultivated Agricultural Areas, Paved Areas 1620.00 1845.02 2880.00 2745.86 Peach 461.25 461.25 720.00 686.46 Pear, Pecan 432.00 492.00 768.00 737.23 Sorghum 54.00 61.50 96.00 91.53 Walnut 425.76 484.9 756.92 721.66 Table 3.5 EECs (ppm) for Indirect Effects to the Terrestrial-Phase CRLF via Effects to Terrestrial Invertebrate Prey Items Use Small Insect Large Insect Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non-Agricultural Areas 2688.80 298.76 Alfalfa, Peppermint, 324.00 36.00 Apple, Grape 504.68 56.08 Citrus, 563.65 62.63 56 ------- Use Small Insect Large Insect Artichoke, Asparagus 432.00 48.00 Banana, Plantain 930.06 103.34 Bermuda grass, Blackberry, Boysenberry, Spearmint 833.22 92.58 Blueberry 239.50 26.61 Field Corn 108.00 12.00 Cotton 452.53 50.28 Dewberry 452.83 50.31 Filbert (Hazelnut) 312.23 34.69 Grass Seed, Wheat 216.00 24.00 Loganberry, Raspberry 324.00 36.00 Olive 223.75 24.86 Pecan, Pear 432.00 48.00 Ornamental Herbaceous plants, Papaya 540.00 60.00 Uncultivated Agricultural Areas, Paved Areas 1620.00 180.00 Peach 461.25 51.25 Sorghum 54.00 6.00 Walnut 425.76 47.31 3.4 Terrestrial Plant Exposure Assessment TerrPlant (version 1.2.2) is used to calculate EECs for non-target plants inhabiting dry and semi-aquatic areas. Parameter values for application rate, drift assumption and incorporation depth are based upon the use and related application method. A runoff value of 0.02 was selected based on diuron's solubility of between 10-100 mg/L. Drift was assumed to be 5% for aerial applications and 1% for ground applications. TerrPlant was used to model EECs from soil and foliar applications relevant to terrestrial plants. These EECs are presented in Table 3.6. An example output from TerrPlant v. 1.2.2 is available in Appendix I. 57 ------- Table 3.6 TerrPlant Inputs and Resulting EECs for Plants Inhabiting Dry and Semi-aquatic Areas Exposed to diuron via Runoff and Drift I se Application rale (lbs a.i./A) Application met hod Drill Value (%) Spray drill i: i :c (lbs a.i./A) l)rv area i: i :c (lbs a.i./A) Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non- Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated N on-Agri cultural Areas 12 Foliar/Ground 0.12 0.36 Alfalfa, Peppermint, Spearmint, Bermudagrass, Blackberry, Boysenberry, Loganberry, Raspberry, Dewberry 2.4 Foliar/Ground 0.02 0.07 Apple, Citrus, Pear, Grape, Artichoke, Asparagus, Pecan 3.2 Foliar/Ground 0.03 0.10 Asparagus (Aerial) 3.2 Foliar/Ground 0.16 0.22 Banana, Plantain 4.8 Foliar/Ground 0.05 0.14 Blueberry, Cotton, Grass Seed, Olive, Wheat 1.6 Foliar/Ground 0.02 0.05 58 ------- I si- Application rale Drill Spray drift l.l.( l)rv area i:i.( Seini- aqualic area i:i.( (lbs a.i./A) Application method Value (%) (lbs a.i./A) (lbs a.i./A) (lbs a.i./A) Field Corn 0.8 Foliar/Ground 1 0.01 0.02 0.17 Filbert (Hazelnut) 2.2 Foliar/Ground 1 0.02 0.07 0.46 Non-agricultural 12 Aerial 5 0.60 0.84 3.00 Ornamental Herbaceous Plants, Papaya 4 Foliar/Ground 1 0.04 0.12 0.84 Uncultivated Agricultural Areas, Paved Areas 12 Foliar/Ground 1 0.12 0.36 2.52 Peach, Walnut 3 Foliar/Ground 1 0.03 0.09 0.63 Sorghum 0.4 Foliar/Ground 1 0.00 0.01 0.08 3.4.1 Spray Drift Buffer Analysis In order to determine terrestrial and aquatic habitats of concern due to diuron exposures through spray drift, it is necessary to estimate the distance that spray applications can drift from the treated area and still be present at concentrations that exceed levels of concern. An analysis of spray drift distances was completed using AgDRIFT and can be found in Table 3.7. AgDRIFT was run for terrestrial analysis only, and due to uncertainties default values were used to determine the aquatic analysis. The default values for PRZM/EXAMS (used for the aquatic analysis) assumed the most conservative scenario, that a buffer was not present. For the terrestrial assessment, the AgDRIFT model was run as a Tier I ground analysis, as well as Tier I, and II aerial analyses focusing on determining a buffer zone for non- listed and listed plant species. The following settings beyond the standard default settings were implemented: • 12 lb ai/A • EC25= 0.002 lb ai/A • ECos = 0.001 lb ai/A • Nonvolatile Rate (Tier II aerial only) = 25 lb/a 59 ------- • Spray Volume Rate (Tier II aerial only) = 3 gal/a • Active Rate (Tier II aerial only) = 4 lb/a The AgDRIFT model was used to evaluate potential distances beyond which exposures would be expected to be below LOC. However, due to the limitations imposed by the Tier 1 ground analysis, and Tiers I and II aerial analyses, which allow users to evaluate off-site deposition and exposure out to 1,000 ft downwind from the location of the application, the exact buffer needed for exposures to be below the LOC is uncertain. Table 3.7 Summary of AgDRIFT Predicted Spray Drift Buffer for Terrestrial Plants 1 iii 1 (iiuiiiMl Applitiilioii Risk Class Risk Description Application Rate Toxicity Value Used Fraction of applied Nonvolatile Rate (lb/a) Minimum Spray Volume Rate (gal/a) Active Rate (ib ai/a) Distance Non- Listed Plants Potential for effects to non- target, non-listed plants from exposures 12 ec25 = 0.002 lb ai/A 0.0002 Does not apply Does not apply Does not apply > 1,000 ft Listed Plants Potential for effects to non- target, listed plants from exposures EC05 = 0.001 lb ai/A 0.0001 1 icr 1 Vcrisil tppliiiilioii Risk Class Risk Description Application Rate Toxicity Value Used Fraction of applied Nonvolatile Rate (lb/a) Minimum Spray Volume Rate (gal/a) Active Rate (ib ai/a) Distance Non- Listed Plants Potential for effects to non- target, non-listed plants from exposures 12 ec25 = 0.002 lb ai/A 0.0002 Does not apply Does not apply Does not apply > 1,000 ft Listed Plants Potential for effects to non- target, listed plants from exposures EC05 = 0.001 lb ai/A 0.0001 Mull Vcrisil \pplit;ili<»ii Risk Class Risk Description Application Rate Toxicity Value Used Fraction of applied Nonvolatile Rate (lb/a) Minimum Spray Volume Rate (gal/a) Active Rate (lb ai/a) Distance Non- Listed Plants Potential for effects to non- target, non-listed plants from exposures 12 ec25 = 0.002 lb ai/A 0.0002 25 3 4 > 1,000 ft Listed Plants Potential for effects to non- target, listed plants from exposures EC05 = 0.001 lb ai/A 0.0001 60 ------- 4.0 Effects Assessment This assessment evaluates the potential for diuron to directly or indirectly affect the CRLF or modify its designated critical habitat. As previously discussed in Section 2.7, assessment endpoints for the CRLF effects determination include direct toxic effects on the survival, reproduction, and growth of CRLF, as well as indirect effects, such as reduction of the prey base or modification of its habitat. In addition, potential modification of critical habitat is assessed by evaluating effects to the PCEs, which are components of the critical habitat areas that provide essential life cycle needs of the CRLF. Direct effects to the aquatic-phase of the CRLF are based on toxicity information for freshwater fish, while terrestrial-phase effects are based on avian toxicity data, given that birds are generally used as a surrogate for terrestrial-phase amphibians. Because the frog's prey items and habitat requirements are dependent on the availability of freshwater fish and invertebrates, small mammals, terrestrial invertebrates, and aquatic and terrestrial plants, toxicity information for these taxa are also discussed. Acute (short- term) and chronic (long-term) toxicity information is characterized based on registrant- submitted studies and a comprehensive review of the open literature on diuron. As described in the Agency's Overview Document (U.S. EPA, 2004), the most sensitive endpoint for each taxon is used for risk estimation. For this assessment, evaluated taxa include aquatic-phase amphibians, freshwater fish, freshwater invertebrates, aquatic plants, birds (surrogate for terrestrial-phase amphibians), mammals, terrestrial invertebrates, and terrestrial plants. Toxicity endpoints are established based on data generated from guideline studies submitted by the registrant, and from open literature studies that meet the criteria for inclusion into the ECOTOX database maintained by EPA/Office of Research and Development (ORD) (U.S. EPA, 2004). Open literature data presented in this assessment were obtained from as well as ECOTOX information obtained on [11/30/2007], In order to be included in the ECOTOX database, papers must meet the following minimum criteria: (1) the toxic effects are related to single chemical exposure; (2) the toxic effects are on an aquatic or terrestrial plant or animal species; (3) there is a biological effect on live, whole organisms; (4) a concurrent environmental chemical concentration/dose or application rate is reported; and (5) there is an explicit duration of exposure. Data that pass the ECOTOX screen are evaluated along with the registrant-submitted data, and may be incorporated qualitatively or quantitatively into this endangered species assessment. In general, effects data in the open literature that are more conservative than the registrant-submitted data are considered. The degree to which open literature data are quantitatively or qualitatively characterized for the effects determination is dependent on whether the information is relevant to the assessment endpoints (i.e., maintenance of CRLF survival, reproduction, and growth) identified in Section 2.8. For example, 61 ------- endpoints such as behavior modifications are likely to be qualitatively evaluated, unless quantitative relationships between modifications and reduction in species survival, reproduction, and/or growth are available. Although the effects determination relies on endpoints that are relevant to the assessment endpoints of survival, growth, or reproduction, it is important to note that the full suite of sublethal endpoints potentially available in the effects literature (regardless of their significance to the assessment endpoints) are considered to define the action area for diuron. Citations of all open literature not considered as part of this assessment because they were either rejected by the ECOTOX screen or accepted by ECOTOX but not used (e.g., the endpoint is less sensitive) are included in Appendix J. This section also includes a rationale for rejection of those studies that did not pass the ECOTOX screen and those that were not evaluated as part of this endangered species risk assessment. A detailed spreadsheet of the available ECOTOX open literature data, including the full suite of lethal and sublethal endpoints is presented in Appendix J. This section also includes a summary of the human health effects data for diuron. In addition to registrant-submitted and open literature toxicity information, other sources of information, including use of the acute probit dose response relationship to establish the probability of an individual effect and reviews of the Ecological Incident Information System (EIIS), are conducted to further refine the characterization of potential ecological effects associated with exposure to diuron. A summary of the available aquatic and terrestrial ecotoxicity information, use of the probit dose response relationship, and the incident information for diuron are provided in Sections 4.1 through 4.4, respectively. The terminal residues of concern in plants and animals are diuron (parent) and metabolites convertible to 3, 4-DCA including desmethoxy-linuron, norlinuron, desmethyl diuron, and hydroxy-norlinuron. The Agency determined that 3, 4-DCA was not of regulatory concern in connection with the registered use of diuron due to the very low levels at which the chemical is detected in plants and animals (<0.01ppm). The Agency concluded that with the possible exception of 3,4-DCA itself, metabolites convertible to 3,4-DCA are not likely to be more toxic than the parent compound. This risk assessment considers the total residues of diuron as parent diuron for purposes of risk assessment A detailed summary of the available ecotoxicity information for all diuron degradates and formulated products are presented in Appendix L. The Agency does not routinely include, in its risk assessments, an evaluation of mixtures of active ingredients, either those mixtures of multiple active ingredients in product formulations or those in the applicator's tank. In the case of the product formulations of active ingredients (that is, a registered product containing more than one active ingredient), each active ingredient is subject to an individual risk assessment for regulatory decision regarding the active ingredient on a particular use site. If effects data 62 ------- are available for a formulated product containing more than one active ingredient, they may be used qualitatively or quantitatively5 6. There are no product LD50 values, with associated 95% Confidence Intervals (CIs) available for diuron. As discussed in USEPA (2000) a quantitative component-based evaluation of mixture toxicity requires data of appropriate quality for each component of a mixture. In this mixture evaluation, an LD50 with associated 95% CI is needed for the formulated product. The same quality of data is also required for each component of the mixture. Given that the formulated products for diuron do not have LD50 data available it is not possible to undertake a quantitative or qualitative analysis for potential interactive effects. However, because the active ingredients are not expected to have similar mechanisms of action, metabolites, or toxicokinetic behavior, it is reasonable to conclude that an assumption of dose-addition would be inappropriate. 4.1 Toxicity of Diuron to Aquatic Organisms Table 4.1 summarizes the most sensitive aquatic toxicity endpoints for the CRLF, based on an evaluation of both the submitted studies and the open literature, as previously discussed. A brief summary of submitted and open literature data considered relevant to this ecological risk assessment for the CRLF is presented below. Additional information is provided in Appendix L. Table 4.1 Freshwater Aquatic Toxicity Profile for Diuron Assessment Endpoint Species Toxicity Value Used in Risk Assessment Citation MRID # or ECOTOX # Comment Acute Direct Toxicity to Aquatic-Phase CRLF Striped Bass (Morone saxatilis) 96h LC50= 400 ppb ECOTOX 102151 Supplemental1 Chronic Direct Toxicity to Aquatic-Phase CRLF Fathead Minnows (Pimephales promelas) NOAEC = 26 ppb ai MRID 00141636 Acceptable (Based on No. of Survivors) Indirect Toxicity to Aquatic-Phase Scud (Gammmarus fasciatus) 48hEC50 = 160 ppb ai MRID 40094602 Acceptable 5 Overview of the Ecological Risk Assessment Process in the Office of Pesticide Programs, Environmental Protection Agency (January 2004) (Overview Document). 6 Memorandum to Office of Prevention, Pesticides and Toxic Substance, US EPA conveying an evaluation by the U.S. Fish and Wildlife Service and National Marine Fisheries Service of an approach to assessing the ecological risks of pesticide products (January 2004). 63 ------- Table 4.1 Freshwater Aquatic Toxicity Profile for Diuron Assessment Endpoint Species Toxicity Value Used in Risk Assessment Citation MRU) # or ECOTOX # Comment CRLF via Acute Toxicity to Freshwater Invertebrates (i.e. prey items) Indirect Toxicity to Aquatic-Phase CRLF via Chronic Toxicity to Freshwater Invertebrates (i.e. prey items) Water Flea (Daphnia magna) 28d NOAEC = 200 ppb ai MRID TN2418 Supplemental1 Indirect Toxicity to Aquatic-Phase CRLF via Acute Toxicity to Non-vascular Aquatic Plants Green Algae (Selenastrum capricornutu m) EC50 = 2.40 ppb ai MRID 42218401 Acceptable Indirect Toxicity to Aquatic-Phase CRLF via Acute Toxicity to Vascular Aquatic Plants Duckweed {Lemna perpusilla) 7d EC50 =15 ppb ECOTOX 8628 Supplemental1 1 This study is considered to be supplemental due to lack of raw data and replicates. Toxicity to aquatic fish and invertebrates is categorized using the system shown in Table 4.2 (U.S. EPA, 2004). Diuron falls in the range of "highly toxic" (0.1 to 1 ppm or 100 to 1000 ppb for fish and invertebrates. Toxicity categories for aquatic plants have not been defined. Table 4.2 Categories of Acute Toxicity for Aquatic Organisms LCso (ppm) Toxicity Category <0.1 Very highly toxic >0.1-1 Highly toxic >1-10 Moderately toxic >10-100 Slightly toxic > 100 Practically nontoxic 64 ------- 4.1.1 Toxicity to Freshwater Fish Given that no diuron toxicity data are available for aquatic-phase amphibians; freshwater fish data were used as a surrogate to estimate direct acute and chronic risks to the CRLF. Freshwater fish toxicity data were also used to assess potential indirect effects of diuron to the CRLF. Effects to freshwater fish resulting from exposure to diuron could indirectly affect the CRLF via reduction in available food. As discussed in Section 2.5.3, over 50% of the prey mass of the CRLF may consist of vertebrates such as mice, frogs, and fish (Hayes and Tennant, 1985). Diuron is highly toxic to both warm water and cold water freshwater fish on an acute exposure basis. The most sensitive freshwater species tested was the warm water Striped Bass species, which exhibited a 96-hour LC50 value of 400 ppb (ECOTOX 102151). Another acute study accepted by the Agency showed the cold water Cutthroat Trout having an acute 96h LC50 of 710 ppb ai (MRID 40098001). The Fathead Minnow exhibited a chronic toxicity NOAEC of 26 ppb based on number of survivors (MRID 00141636). 4.1.2 Toxicity to Freshwater Invertebrates Freshwater aquatic invertebrate toxicity data were used to assess potential indirect effects of diuron to the CRLF. Effects to freshwater invertebrates resulting from exposure to diuron could indirectly affect the CRLF via reduction in available food items. As discussed in Section 2.5.3, the main food source for juvenile aquatic- and terrestrial- phase CRLFs is thought to be aquatic invertebrates found along the shoreline and on the water surface, including aquatic sowbugs, larval alderflies and water striders. Diuron is highly acutely toxic to freshwater invertebrates based on acceptable studies on the Scud. This species exhibited a 48-hour EC50 value of 160 ppb (MRID: 40094602). Chronic toxicity studies showed the Water Flea as exhibiting a 28d NOAEC of 200 ppb (MRID: TN2418). 4.1.3 Toxicity to Aquatic Plants Aquatic plant toxicity studies were used as one of the measures of effect to evaluate whether diuron may affect primary production and the availability of aquatic plants as food for CRLF tadpoles. Primary productivity is essential for indirectly supporting the growth and abundance of the CRLF. Two types of studies were used to evaluate the potential of diuron to affect aquatic plants. Laboratory and field studies were used to determine whether diuron may cause direct effects to aquatic plants. A summary of the laboratory data and freshwater field studies for aquatic plants is provided in Sections 4.1.3.1 and 4.1.4. The Green Alga Selenastrum capricornutum exhibited the most sensitive non-vascular plant acute endpoint with an EC50 of 2.40 ppb. The aquatic vascular plant Lemna 65 ------- perpusilla exhibited the most sensitive vascular acute endpoint with a 7d EC50 of 15 ppb ai (ECOTOX 8628). 4.2 Toxicity of Diuron to Terrestrial Organisms Table 4.3 summarizes the most sensitive terrestrial toxicity endpoints for the CRLF, based on an evaluation of both the submitted studies and the open literature. A brief summary of submitted and open literature data considered relevant to this ecological risk assessment for the CRLF is presented below. Table 4.3 Terrestrial Toxicity Profile for Diuron Endpoint Species Toxicity Value Used in Risk Assessment Citation MR ID# Comment Acute Direct Toxicity to Terrestrial- Phase CRLF (LD50) Mallard Duck (Anas platyrhynchos) 14d LD50 =>2000 mg/kg-bw MRID 00160000 Acceptable No mortality reported. Ataxia persisted for 1 Id after study Acute Direct Toxicity to Terrestrial- Phase CRLF (LC50) Northern Bobwhite Quail (Colinus virgianus) 9d LC50 = 1730 ppm ai MRID 00022923 Acceptable Chronic Direct Toxicity to Terrestrial- Phase CRLF No Acceptable Study Available Indirect Toxicity to Terrestrial- Phase CRLF (via acute toxicity to mammalian prey items) Laboratory Rat (Rattus norvegieus) LD50 - <$ 5000 mg/kg-bw $ 10000 mg/kg-bw MRID 00146145 Acceptable Indirect Toxicity to Terrestrial- Phase CRLF (via chronic toxicity to mammalian prey items) Laboratory Rat (Rattus norvegieus) NOAEC = 250 ppm MRID 00146145 Acceptable Indirect Toxicity to Terrestrial- Phase CRLF (via acute toxicity to terrestrial invertebrate prey items) Honey Bee (Apis mellifera) LD50 >145 (ig/bee (1133.05 ppm) MRID 00036935 Acceptable (Only 2.7% mortality observed at highest level tested) Indirect Dicot Seedling EC-h = 0.075 lb ai/A MRID Acceptable 66 ------- Endpoint Species Toxicity Value Used in Risk Assessment Citation MRID# Comment Emergence Tomato 44113401 (Lycopersicon esculentum) Toxicity to Terrestrial- Dicot Vegetative Vigor Tomato (.Lycopersicon esculentum) EC25 = 0.002 lb ai/A MRID 44113401 Acceptable and Aquatic- Phase CRLF Monocot (via toxicity to terrestrial plants) Seedling Emergence Onion (Allium cepa) EC25 = 0.099 lb ai/A MRID 44113401 Acceptable Monocot Vegetative Vigor Wheat (Triticum aestivum) EC25 = 0.021 lb ai/A MRID 44113401 Acceptable Acute toxicity to terrestrial animals is categorized using the classification system shown in Table 4.4 (U.S. EPA, 2004). Toxicity categories for terrestrial plants have not been defined. Table 4.4 Categories of Acute Toxicity for Avian and Mammalian Studies Toxicity Category Oral LDsn Dietary LC50 Very highly toxic <10 mg/kg < 50 ppm Highly toxic 10-50 mg/kg 50 - 500 ppm Moderately toxic 51 - 500 mg/kg 501 - 1000 ppm Slightly toxic 501 - 2000 mg/kg 1001 - 5000 ppm Practically non-toxic > 2000 mg/kg > 5000 ppm 4.2.1 Toxicity to Birds As specified in the Overview Document, the Agency uses birds as a surrogate for terrestrial-phase amphibians when amphibian toxicity data are not available (U.S. EPA, 2004). No terrestrial-phase amphibian data are available for diuron; therefore, acute and chronic avian toxicity data are used to assess the potential direct effects of diuron to terrestrial-phase CRLFs. There are no data by which to assess chronic toxicity to avian species (surrogates for the terrestrial phase frog) either submitted by the registrants or in the public literature. Linuron, a similar chemical of the same class, has acceptable chronic avian data. The avian reproductive studies for linuron (MRID 42541801, 42541802) using mallard duck and bobwhite quail found reproductive effects at 300 ppm ai with an NOAEL being established at 100 ppm. The endpoints affected for mallard duck reproduction are egg production, adult body weight, feed consumption, viable embryos of eggs set, number of viable embryos, and number of live embryos. For the bobwhite quail the endpoints 67 ------- affected are hatchability and offspring survivability. This suggests that diuron may have chronic effects to avian species, but these risks cannot be quantified in the absence of data. 4.2.2 Toxicity to Mammals Mammalian toxicity data are used to assess potential indirect effects of diuron to the terrestrial-phase CRLF. Effects to small mammals resulting from exposure to diuron could also indirectly affect the CRLF via reduction in available food. As discussed in Section 2.5.3, over 50% of the prey mass of the CRLF may consist of vertebrates such as mice, frogs, and fish (Hayes and Tennant, 1985). An acute oral LD50 of 5000 mg/kg-bw was observed in the Laboratory Rat (MRID 00146145). The Laboratory Rat NOAEC is 250 ppm (MRID 00146145). 4.2.3 Toxicity to Terrestrial Invertebrates Terrestrial invertebrate toxicity data are used to assess potential indirect effects of diuron to the terrestrial-phase CRLF. Effects to terrestrial invertebrates resulting from exposure to diuron could also indirectly affect the CRLF via reduction in available food. Diuron is classified as practically non-toxic to the Honey Bee. The acute LD50 for Honey Bee is >145.03 |ig/bee (MRID 00036935). Only 2.7% mortality was observed in this study at the highest level tested. The test was therefore not definitive and RQs were not calculated. There were no additional acceptable terrestrial invertebrate data from registrant submitted studies or from the open literature by which to calculate RQ values. 4.2.4 Toxicity to Terrestrial Plants Terrestrial plant toxicity data are used to evaluate the potential for diuron to affect riparian zone and upland vegetation within the action area for the CRLF. Impacts to riparian and upland (i.e., grassland, woodland) vegetation could result in indirect effects to both aquatic- and terrestrial-phase CRLFs, as well as modification to designated critical habitat PCEs via increased sedimentation, alteration in water quality, and reduction in of upland and riparian habitat that provides shelter, foraging, predator avoidance and dispersal for juvenile and adult CRLFs. Plant toxicity data from both registrant-submitted studies and studies in the scientific literature were reviewed for this assessment. Registrant-submitted studies are conducted under conditions and with species defined in EPA toxicity test guidelines. Sub-lethal endpoints such as plant growth, dry weight, and biomass are evaluated for both monocots and dicots, and effects are evaluated at both seedling emergence and vegetative life stages. Guideline studies generally evaluate toxicity to ten crop species. A drawback to these tests is that they are conducted on herbaceous crop species only, and extrapolation of effects to other species, such as the woody shrubs and trees and wild herbaceous species, contributes uncertainty to risk conclusions. 68 ------- Commercial crop species have been selectively bred, and may be more or less resistant to particular stressors than wild herbs and forbs. The direction of this uncertainty for specific plants and stressors, including diuron, is largely unknown. Homogenous test plant seed lots also lack the genetic variation that occurs in natural populations, so the range of effects seen from tests is likely to be smaller than would be expected from wild populations. The most sensitive endpoints reported for diuron exposure to dicot plants are a vegetative vigor EC25 value of 0.002 lb ai/A and a seedling emergence EC25 value of 0.075 lb ai/A as expressed by tomato plants (MRID 44113401). The most sensitive endpoints reported for diuron exposure to monocot plants are a vegetative vigor EC25 value of 0.021 lb ai/A as expressed by Wheat, and a seedling emergence EC25 value of 0.099 lb ai/A as expressed by Onion (MRID 44113401). The results of the Tier II seedling emergence and vegetative vigor toxicity tests on non-target plants are summarized below in Table 4.5. Table 4.5 Terrestrial Plant Toxicity Profile for Diuron Endpoint Species Toxicity Value Used in Risk Assessment Citation MRID# Comment Dicot Seedling Emergence Tomato (Lycopersicon esculentum) EC25 = 0.075 lb ai/A MRID 44113401 Acceptable Indirect Toxicity to Terrestrial- and Aquatic- Dicot Vegetative Vigor Tomato (.Lycopersicon esculentum) EC25 = 0.002 lb ai/A MRID 44113401 Acceptable Phase CRLF (via toxicity to terrestrial plants) Monocot Seedling Emergence Onion (Allium cepa) EC25 = 0.099 lb ai/A MRID 44113401 Acceptable Monocot Vegetative Vigor Wheat (Triticum aestivum) EC25 = 0.021 lb ai/A MRID 44113401 Acceptable 4.3 Use of Probit Slope Response Relationship to Provide Information on the Endangered Species Levels of Concern The Agency uses the probit dose response relationship as a tool for providing additional information on the potential for acute direct effects to individual listed species and aquatic animals that may indirectly affect the listed species of concern (U.S. EPA, 2004). As part of the risk characterization, an interpretation of acute RQ for listed species is discussed. This interpretation is presented in terms of the chance of an individual event (i.e., mortality or immobilization) should exposure at the EEC actually occur for a species with sensitivity to diuron on par with the acute toxicity endpoint selected for RQ calculation. To accomplish this interpretation, the Agency uses the slope of the dose 69 ------- response relationship available from the toxicity study used to establish the acute toxicity measures of effect for each taxonomic group that is relevant to this assessment. The individual effects probability associated with the acute RQ is based on the mean estimate of the slope and an assumption of a probit dose response relationship. In addition to a single effects probability estimate based on the mean, upper and lower estimates of the effects probability are also provided to account for variance in the slope, if available. Individual effect probabilities are calculated based on an Excel spreadsheet tool IECV1.1 (Individual Effect Chance Model Version 1.1) developed by the U.S. EPA, OPP, Environmental Fate and Effects Division (June 22, 2004). The model allows for such calculations by entering the mean slope estimate (and the 95% confidence bounds of that estimate) as the slope parameter for the spreadsheet. In addition, the acute RQ is entered as the desired threshold. 4.4 Incident Database Review A review of the EIIS database for ecological incidents involving diuron was completed on 12/18/08. Thirty highly probable, probable or possible incidents involving diuron have been reported. These uses were either registered uses or undetermined uses. One of these incidents resulted in a bird kill, 22 resulted in plant damage and seven resulted in fish kills. A detailed description of incidents relating to diuron use is presented in Appendix M. These incidents will be used in addition to other lines of evidence to draw conclusions regarding the risks of diuron to the CRLF. The absence of additional documented incidents does not necessarily mean that such incidents did not occur. Mortality incidents must be seen, reported, investigated, and submitted to the Agency in order to be recorded in the incident database. Incidents may not be noticed because the carcasses decayed, were removed by scavengers, or were in out-of-the-way or hard-to-see locations. Due to the voluntary nature of incident reporting, an incident may not be reported to appropriate authorities capable of investigating it. 5.0 Risk Characterization Risk characterization is the integration of the exposure and effects characterizations. Risk characterization is used to determine the potential for direct and/or indirect effects to the CRLF or for modification to its designated critical habitat from the use of diuron in CA. The risk characterization provides an estimation (Section 5.1) and a description (Section 5.2) of the likelihood of adverse effects; articulates risk assessment assumptions, limitations, and uncertainties; and synthesizes an overall conclusion regarding the likelihood of adverse effects to the CRLF or its designated critical habitat (i.e., "no effect," "likely to adversely affect," or "may affect, but not likely to adversely affect"). 5.1 Risk Estimation Risk is estimated by calculating the ratio of exposure to toxicity. This ratio is the risk quotient (RQ), which is then compared to pre-established acute and chronic levels of 70 ------- concern (LOCs) for each category evaluated (Appendix E). For acute exposures to the CRLF and its animal prey in aquatic habitats, as well as terrestrial invertebrates, the LOC is 0.05. For acute exposures to the CRLF and mammals, the LOC is 0.1. The LOC for chronic exposures to CRLF and its prey, as well as acute exposures to plants is 1.0. Risk to the aquatic-phase CRLF is estimated by calculating the ratio of exposure to toxicity using l-in-10 year EECs based on the label-recommended diuron usage scenarios summarized in Table 3.2 and the appropriate aquatic toxicity endpoint from Table 4.1. Risks to the terrestrial-phase CRLF and its prey (e.g. terrestrial insects, small mammals and terrestrial-phase frogs) are estimated based on exposures resulting from applications of diuron (Tables 3.5 through 3.6) and the appropriate toxicity endpoint from Table 4.6. Exposures are also derived for terrestrial plants, as discussed in Section 3.3 and summarized in Table 3.7, based on the highest application rates of diuron use within the action area. 5.1.1 Exposures in the Aquatic Habitat 5.1.1.1 Direct Effects to Aquatic-Phase CRLF Direct effects to the aquatic-phase CRLF are based on peak EECs in the standard pond and the lowest acute toxicity value for freshwater fish. In order to assess direct chronic risks to the CRLF, 60-day EECs and the lowest chronic toxicity value for freshwater fish are used. The RQs for diuron uses results in acute and chronic exceedances of the Agency's LOC for freshwater fish which are surrogates for the aquatic phase for amphibians. The aquatic phase amphibian acute LOCs for listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 12.28 (paved areas) to 0.06 (papaya and walnut). Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 131.85 (paved areas) to 1.26 (banana, plantain). A probit slope value for the acute fathead minnow toxicity test is not available; therefore, the individual effect probability was estimated based on a default slope assumption of 4.5 with upper and lower 95% confidence intervals of 2 and 9 (Urban and Cook, 1986). The estimated probability of an individual effect for treatments that exceeded the Agency's acute RQs for listed species ranged from approximately 1 in 1 for paved areas to approximately 1 in 1.64 E+4 for banana/plantain (upper and lower 95% confidence intervals of 1 in 2.28 E+01 to 1 in 1.29 E+14). Based on these exceedances, diuron May Affect the aquatic-phase of the CRLF. Results are presented in Table 5 .1. 71 ------- Table 5.1. Summary of Direct Effect RQs for the Aquatic-phase CRLF l si- Diivii 1'. MVils In C kl.l ' SuiTii»;ik- S|K-iii-s Tn\ii'ii> Value II. ( IV;ik/fiOil RQ Pnihiihililx 11I' liuli\ iduiil l.lliii 1 l.(H r.ui'i'ikiiui- iind Risk liiu-rpivliiliiin AGR1CLLTLR.IL R1GHTS-OF- WA Y/FENCERO WS/ HEDGEROWS Aailc Direct Toxicity Si II pod Bass LCJV 400 l.X" 1.72 1 1111.1" (.1 in 1.47 to 1 in 1.07)b Ycsc Chronic Direct Toxicity Fathead Minnow NOAEC = 26 456.81 17.57 Not applicable for chronic endpoints Yes'1 ALFALFA Acute Direct Toxicity Striped Bass lc50 = 400 10.63 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 7.20 0.28 Not applicable for chronic endpoints No ALFALFA (Aerial) Acute Direct Toxicity Striped Bass LC50 = 400 13.69 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 9.40 0.36 Not applicable for chronic endpoints No APPLE Acute Direct Toxicity Striped Bass LC50 = 400 11.68 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 7.80 0.30 Not applicable for chronic endpoints No ARTLCHOKE Acute Direct Toxicity Striped Bass LC50 = 400 32.00 0.08 1 in 2.51E+06 (1 in 1.00 E+16 to 1 in 7.08 E+l)b Yes' 72 ------- Chronic Direct Toxicity Fathead Minnow NOAEC = 26 25.08 0.96 Not applicable for chronic endpoints No ASPARGUS Acute Direct Toxicity Striped Bass LC50 = 400 36.99 0.09 1 in 7.90 E+05 (1 in 1.00 E+16 to 1 in 1.00 E+06)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 22.92 0.88 Not applicable for chronic endpoints No BANANA, PLANTAIN Acute Direct Toxicity Striped Bass LC50 = 400 54.21 0.14 1 in 1.64 E+4 (1 in 2.28 E+01 to 1 in 1.29 E+14)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 32.78 1.26 Not applicable for chronic endpoints Yes" BLACKBERRY, BOYSENBERRY Acute Direct Toxicity Striped Bass LC50 = 400 139.70 0.35 1 in 4.98E+01 (1 in 5.53 to 1 in 4.91 E+4)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 102.90 3.96 Not applicable for chronic endpoints Yes4 BLUEBERRY Acute Direct Toxicity Striped Bass LC50 = 400 4.65 0.01 <1 in 1.00E+16 (1 in 3.16 E+04 to <1 in 1.00E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 3.21 0.12 Not applicable for chronic endpoints No CITRUS Acute Direct LC50 = 400 6.83 0.02 1 in 9.48 E+13 (1 in No 73 ------- Toxicity 2.95 E+3 to <1 in 1.00 E+16)b Chronic Direct Toxicity Rainbow Trout NOAEC = 26 4.88 0.19 Not applicable for chronic endpoints No CORN, FIELD Acute Direct Toxicity Striped Bass LC50 = 400 7.24 0.02 1 in 9.48 E+13 (1 in 2.95 E+3 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 4.68 0.18 Not applicable for chronic endpoints No DEWBERRY Acute Direct Toxicity Striped Bass LC50 = 400 19.71 0.05 1 in 4.17 E+08 (1 in 2.16 E+02 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 14.29 0.55 Not applicable for chronic endpoints No FILBERT (HAZELNUT) Acute Direct Toxicity Striped Bass LC50 = 400 18.90 0.05 1 in 4.17 E+08 (1 in 2.16 E+02 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 12.16 0.47 Not applicable for chronic endpoints No GRAPE Acute Direct Toxicity Striped Bass LC50 = 400 9.71 0.02 1 in 9.48 E+13 (1 in 2.95 E+3 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 6.37 0.24 Not applicable for chronic endpoints No LOGANBERRY, RASPBERRY Acute Direct Striped Bass LC50 = 400 5.13 0.01 <1 in 1.00E+16 (1 No 74 ------- (BLACK/RED) Toxicity in 3.16 E+04 to <1 in 1.00E+16)b Chronic Direct Toxicity Fathead Minnow NOAEC = 26 3.02 0.12 Not applicable for chronic endpoints No OLIVE Acute Direct Toxicity Striped Bass LC50 = 400 7.17 0.02 1 in 9.48 E+13 (1 in 2.95 E+3 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 4.86 0.19 Not applicable for chronic endpoints No PAPAYA Acute Direct Toxicity Striped Bass LC50 = 400 25.21 0.06 1 in 5.20 E+07 (1 in 1.38 E+02 to <1 in 1.00 E+16)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 15.72 0.60 Not applicable for chronic endpoints No PEPPERMINT Acute Direct Toxicity Striped Bass LC50 = 400 4.17 0.01 <1 in 1.00E+16 (1 in 3.16 E+04 to <1 in 1.00E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 2.79 0.11 Not applicable for chronic endpoints No PEACH Acute Direct Toxicity Striped Bass LC50 = 400 10.95 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 7.31 0.28 Not applicable for chronic endpoints No PEAR Acute Direct Toxicity Striped Bass LC50 = 400 11.66 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No 75 ------- Chronic Direct Toxicity Fathead Minnow NOAEC = 26 7.79 0.30 Not applicable for chronic endpoints No PECAN Acute Direct Toxicity Striped Bass LC50 = 400 27.04 0.07 1 in 9.86 E+06 (1 in 9.57 E+l to <1 in 1.00 E+16)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 17.33 0.67 Not applicable for chronic endpoints No SORGHUM Acute Direct Toxicity Striped Bass LC50 = 400 5.87 0.01 <1 in 1.00E+16 (1 in 3.16 E+04 to <1 in 1.00E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 4.52 0.17 Not applicable for chronic endpoints No SPEARMINT Acute Direct Toxicity Striped Bass LC50 = 400 106.70 0.27 1 in 1.90E+02 (1 in 7.83 to 1 in 6.46 E+06)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 81.95 3.15 Not applicable for chronic endpoints Yesd WALNUT (ENGLISH/BLACK) Acute Direct Toxicity Striped Bass LC50 = 400 25.78 0.06 1 in 5.20 E+07 (1 in 1.38 E+02 to <1 in 1.00 E+16)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 16.58 0.64 Not applicable for chronic endpoints No WHEAT (pre- harvest) Acute Direct Toxicity Striped Bass LC50 = 400 18.03 0.05 1 in 4.17 E+08 (1 in 2.16 E+02 to <1 in 1.00 E+16)b No 76 ------- Chronic Direct Toxicity Fathead Minnow NOAEC = 26 12.02 0.46 Not applicable for chronic endpoints No WHEAT (post- harvest) Acute Direct Toxicity Striped Bass LC50 = 400 12.82 0.03 1 in 2.75E+11 (1 in 1.00 E+16 to 1 in 8.63 E+02)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 10.29 0.40 Not applicable for chronic endpoints No COTTON (ground) Acute Direct Toxicity Striped Bass LC50 = 400 8.36 0.02 1 in 9.48 E+13 (1 in 2.95 E+3 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 5.72 0.22 Not applicable for chronic endpoints No COTTON (aerial) Acute Direct Toxicity Striped Bass LC50 = 400 14.25 0.04 1 in 6.30 E+09 (1 in 3.86 E+02 to <1 in 1.00 E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 10.24 0.39 Not applicable for chronic endpoints No AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS Acute Direct Toxicity Striped Bass LC50 = 400 2190.00 5.48 ~1 in lb Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 1382.00 53.15 Not applicable for chronic endpoints Yes" BERMUDAGRASS Acute Direct Toxicity Striped Bass LC50 = 400 226.00 0.57 1 in 7.35 (1 in 3.20 to 1 in 7.14 E+01)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 167.90 6.46 Not applicable for chronic endpoints Yes'1 77 ------- GRASSES GROWN FOR SEED Acute Direct Toxicity Striped Bass £ r o II 3.76 0.01 <1 in 1.00E+16 (1 in 3.16 E+04 to <1 in 1.00E+16)b No Chronic Direct Toxicity Fathead Minnow NOAEC = 26 2.89 0.11 Not applicable for chronic endpoints No IRRIGATION SYSTEMS Acute Direct Toxicity Striped Bass LC50 = 400 52.56 0.13 1 in 2.99E+04 (1 in 2.62 E+01 to 1 in 1.29 E+15)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 37.45 1.44 Not applicable for chronic endpoints Yes' ORNAMENTAL HERBACEOUS PLANTS Acute Direct Toxicity Striped Bass LC50 = 400 36.55 0.09 1 in 7.90 E+05 (1 in 5.48 E+01 to <1 in 1.00 E+16)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 23.56 0.91 Not applicable for chronic endpoints No NON- AGRICULTURAL RIGHTS OF WAY (ground) Acute Direct Toxicity Striped Bass LC50 = 400 629.91 1.57 1 in 1.23.(1 in 1.53 to 1 in 1.04)b Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 476.35 18.32 Not applicable for chronic endpoints Yes' NON- AGRICULTURAL RIGHTS OF WAY (aerial) Acute Direct Toxicity Striped Bass LC50 = 400 626.91 1.57 1 in 1.23.(1 in 1.53 to 1 in 1.04)b Yes* Chronic Direct Toxicity Fathead Minnow NOAEC = 26 468.16 18.01 Not applicable for chronic endpoints Yesd PAVED AREAS Acute Direct Toxicity Striped Bass LC50 = 400 4911.00 12.28 ~1 in lb Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 3428.00 131.85 Not applicable for chronic endpoints Yes' 78 ------- UNCULTIVATED AG Acute Direct Toxicity Striped Bass £ r o II 1130.00 2.83 ~1 in lb Yes' Chronic Direct Toxicity Fathead Minnow NOAEC = 26 840.40 32.32 Not applicable for chronic endpoints Yesd UNCULTIVATED NON-AG Acute Direct Toxicity Striped Bass LC50 = 400 284.80 0.71 1 in 3.97(1 in 2.61 to 1 in 1.11 E+01)b Yes1 Chronic Direct Toxicity Fathead Minnow NOAEC = 26 202.50 7.79 Not applicable for chronic endpoints Yes' a RQs associated with acute and chronic direct toxicity to the CRLF are also used to assess potential indirect effects to the CRLF based on a reduction in freshwater fish and frogs as food items. b A probit slope value for the acute fathead minnow toxicity test is not available; therefore, the effect probability was calculated based on a default slope assumption of 4.5 with upper and lower 95% confidence intervals of 2 and 9 (Urban and Cook, 1986). c RQ > acute endangered species LOC of 0.05. dRQ> chronic LOC of 1.0. 5.1.1.2 Indirect Effects to Aquatic-Phase CRLF via Reduction in Prey (non-vascular aquatic plants, aquatic invertebrates, fish, and frogs) Non-vascular Aquatic Plants Indirect effects of diuron to the aquatic-phase CRLF (tadpoles) via reduction in non- vascular aquatic plants in its diet are based on peak EECs from the standard pond and the lowest acute toxicity value for aquatic non-vascular plants. The Agency's LOC (1.0) is exceeded for all current uses of diuron in California. The RQs range from 2046.25 (paved areas) to 1.57 (grasses grown for seed). Results are presented in Table 5.2. Based on these exceedances, diuron May Affect the CRLF via reduction of non- vascular plants. 79 ------- Table 5.2 Summary of Acute RQs Used to Estimate Indirect Effects to the CRLF via Effects to Non-Vascular Aquatic Plants (diet of CRLF in tadpole life stage and habitat of aquatic-phase CR LF)* l SI'S Applkiilinn nili- (II) ;ii/.\) A|)|)lii;ili(ili Mi-lhml IY;ik I.I. ( (ii»/l.) liidiivil i-ITi-ils RQ : (I'ikkI iiiul hiihiiiii) 2N(i.50 4.43 AGRICULTURAL RIGHTS-OF- WAY/FENCEROW S/HEDGEROW S ALFALFA 12 Ground 687.59 2.4 Ground 10.63 2.4 Aircraft 13.69 5.71 APPLE 3.2 Ground 11.68 4.S7 ARTICHOKE 3.2 Ground 32.00 13.33 ASPARGUS 4.8 Ground 36.99 15.41 BANANA, PLANTAIN 2.4 Ground 54.21 22.5'> BLACKBERRY, BOYSENBERRY 1.6 Ground 139.70 5S.2I BLUEBERRY 2.4 Ground 4.65 l.'M CITRUS 3.2 Ground 6.83 2.S5 CORN, FIELD 0.8 Ground 7.24 3.02 DEWBERRY 2.4 Ground 19.71 S.2I FILBERT (HAZELNUT) 2.2 Ground 18.90 7.87 80 ------- l SI'S .\|>|>lk;ilii>ll nili- (II) ;ii/.\) A|)|)lii;ili(ili \klhml IY;ik I.I. ( (iiS/l.) liiriiivil i-fli-ils RQ : (I'm id ;mkI li;il)iliil) 4.04 GRAPE 3.2 Ground 9.71 LOGANBERRY, RASPBERRY (BLACK/RED) 2.4 Ground 5.13 2.14 OLIVE 1.6 Ground 7.17 2.')') PAPAYA 4 Ground 25.21 10.50 PEPPERMINT 2.4 Ground 4.17 1.74 PEACH 3 Ground 10.95 4.5f. PEAR 3.2 Ground 11.66 4.Sfi PECAN 3.2 Ground 27.04 11.27 SORGHUM 4.8 Ground 5.87 2.45 SPEARMINT 2.4 Ground 106.70 44.4r. WALNUT (ENGLISH/BLACK) 0.4 Ground 25.78 10.74 WHEAT 2.4 Pre-Harvest 18.03 7.51 3 Post- Harvest 12.82 5.34 COTTON 1.6 Ground 8.36 3.4S Aircraft 14.25 5.'M 81 ------- l SI'S .\|>|>lk;ilii>ll nili- (II) ;ii/.\) A|)|)lii;ili(ili \klhml IY;ik I.I. ( (iiS/l.) liiriiivil i-ITi-ils RQ : (I'm id ;mkI li;il)iliil) •>12.50 AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS 12 Ground 2190.00 BERMUDAGRASS 2.4 Ground 226.00 'M.I 7 GRASSES GROWN FOR SEED 12 Ground 3.76 1.57 IRRIGATION SYSTEMS 1.6 Ground 52.56 2I.'>0 ORNAMENTAL HERBACEOUS PLANTS 12 Ground 36.55 15.23 NON-AGRICULTURAL RIGHTS OF WAY 12 Ground 629.91 2r.2.4r. 4 Aircraft 626.91 261.21 PAVED AREAS (PRIVATE ROADS/SIDEWALKS) 12 Ground 4911.00 2IW6.25 UNCULTIVATED AG 12 Ground 1130.00 470.S3 4S7.0S 12 Ground 1169.00 UNCULTIVATED NON-AG 12 Ground 284.80 11 S.f.7 82 ------- l SI'S Applkiilinn nili- (II) ;ii/.\) Application Milliml IY;ik I.I. ( (ii»/l.) Indiri-il i-lli-ils RQ- (I'm id ;mkI h;il)iliil) * LOC exceedances (RQ > 1.0) are bolded and shaded. RQ = use-specific peak EEC/ [Selenastrum capricornutum EC50 = 2.40 ppb]. Aquatic Invertebrates Indirect acute effects to the aquatic-phase CRLF via effects to prey (invertebrates) in aquatic habitats are based on peak EECs in the standard pond and the lowest acute toxicity value for freshwater invertebrates. For chronic risks, 21-day EECs and the lowest chronic toxicity value for invertebrates are used to derive RQs. The Agency's acute Listed LOC (0.05) is exceeded for most uses of diuron in California. The acute RQs that exceed the LOC range from 30.69 (Paved Areas) to 0.06 (grape). The chronic RQs exceed the Agency's LOC for several used. The chronic values that exceeded range from 21.21 (Paved Areas) to 1.02 (Bermudagrass). A summary of the acute and chronic RQ values for exposure to aquatic invertebrates (as prey items of aquatic-phase CRLFs) is provided in Table 5.3. Based on acute Listed LOC exceedances for chronic and acute aquatic invertebrates from most use sites, diuron May Affect the CRLF indirectly via reduction in freshwater invertebrate prey items. Table 5.3 Summary of Acute and Chronic RQs Used to Estimate Indirect Effects to the CRLF via Direct Effects on Aquatic Invertebrates as Dietary Food Items (prey of CRLF juveniles and adults in aquatic habitats)* I si-s Applkiilinn l llll' (II) ;ii/A) A|)|)lii;iliiin Mi-llmd IY;ik l.l.t (iiii/l-l 21 l);i> I.I. ( |)|)l> lndiri-il IHTi'iis Aiulc RQ Indin-ii 1". ITi-ils Cli ii in i i' RQ AGRICULTURAL RIGHTS-OF- WAY/FENCEROW S/HEDGEROW S 12 Ground 687.59 672.03 4.30 2.'>2 ALFALFA 2.4 Ground 10.63 10.37 imp 005 2.4 Aircraft 13.69 13.29 o.oy 83 ------- APPLE 3.2 Ground 11.68 11.22 0.07 0.05 ARTICHOKE 3.2 Ground 32.00 31.15 0.20 0.14 ASPARGUS 4.8 Ground 36.99 35.46 0.23 0.15 BANANA, PLANTAIN 2.4 Ground 54.21 51.95 0.34 0.22 BLACKBERRY, BOYSENBERRY 1.6 Ground 139.70 135.80 O.S^ 0.60 BLUEBERRY 2.4 Ground 4.65 4.48 0.03 0.02 CITRUS 3.2 Ground 6.83 6.57 0.04 0.03 CORN, FIELD 0.8 Ground 7.24 6.95 0.05 0.03 DEWBERRY 2.4 Ground 19.71 19.25 0.12 0.08 FILBERT (HAZELNUT) 2.2 Ground 18.90 18.41 0.12 0.08 GRAPE 3.2 Ground 9.71 9.32 0.0(> 0.04 LOGANBERRY, RASPBERRY (BLACK/RED) 2.4 Ground 5.13 4.95 0.03 0.02 OLIVE 1.6 Ground 7.17 6.90 0.04 0.03 PAPAYA 4 Ground 25.21 24.56 O.I(> 0.11 PEPPERMINT 2.4 Ground 4.17 4.05 1)0' 0.02 PEACH 3 Ground 10.95 10.51 0.07 0.05 PEAR 3.2 Ground 11.66 11.20 0.0"7 0.05 PECAN 3.2 Ground 27.04 26.33 o.r 0.11 SORGHUM 4.8 Ground 5.87 5.67 0.04 0.03 SPEARMINT 2.4 Ground 106.70 103.70 O.h"7 0.49 84 ------- WALNUT (ENGLISH/BLACK) 0.4 Ground 25.78 25.11 0. Hi 0.11 WHEAT 2.4 Pre-Harvest 18.03 17.34 0.1 1 1 I.I IS 0.07 3 Post- Harvest 12.82 12.34 0.06 COTTON 1.6 Ground 8.36 8.13 0.05 0.04 Aircraft 14.25 13.88 0.1(9 IKK. AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS 12 Ground 2190.00 2115.00 1 '>.13 BERMUDAGRASS 2.4 Ground 226.00 218.80 1.41 1.02 GRASSES GROWN FOR SEED 12 Ground 3.76 3.62 0.02 0.02 IRRIGATION SYSTEMS 1.6 Ground 52.56 50.67 0.33 u: ^ ORNAMENTAL HERBACEOUS PLANTS 12 Ground 36.55 35.45 0.23 u 15 NON-AGRICULTURAL RIGHTS OF WAY 12 Ground 629.91 619.12 3.9 4 2.S0 4 Aircraft 626.91 615.83 3.') 2 2.7(, PAVED AREAS (PRIVATE ROADS/SIDEWALKS) 12 Ground 4911.00 4725.00 30.6') 21.21 UNCULTIVATED AG 12 Ground 1130.00 1099.00 7.0(. 5.12 12 Ground 1169.00 1130.00 7.3I 5.31 UNCULTIVATED NON-AG 12 Ground 284.80 275.70 I.7X 1.2ft 85 ------- * LOC exceedances (acute RQ > 0.05, chronic RQ > 1.0) are bolded and shaded. RQ = use-specific peak EEC/ [Selenastrum capricornutum EC50 = 2.40 ppb]. Fish and Frogs Fish and frogs also represent potential prey items of adult aquatic-phase CRLFs. RQs associated with acute and chronic direct toxicity to the CRLF (Table 5.1) are used to assess potential indirect effects to the CRLF based on a reduction in freshwater fish and frogs as food items. Based on chronic and acute LOC exceedances, diuron May Affect the CRLF indirectly via reduction in freshwater fish and frogs as food items. 5.1.1.3 Indirect Effects to CRLF via Reduction in Habitat and Primary Productivity (Freshwater Aquatic Plants) Indirect effects to the CRLF via direct toxicity to aquatic plants are estimated using the most sensitive non-vascular and vascular plant toxicity endpoints. A summary of acute RQs used to estimate indirect effects to the CRLF via effects to vascular aquatic plants is presented in Table 5.4. The RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses of diuron in California. These range from 327.40 (paved areas) to 1.26 (Filbert/Hazelnut). Because of this, and in addition to the LOC exceedances for all uses of diuron to non-vascular aquatic plants, it is determined that diuron May Affect the CRLF indirectly via reduction in vascular aquatic plants. Table 5.4 Summary of Acute RQs Used to Estimate Indirect Effects to the CRLF via I SI'S r;ik- (II) ;ii/.\) Applkiiliiiii Mi-lhml IY;ik II. ( (ii»/l.) Indiivi'l l-'.ITi'i'ls Auik- RQ* AGRICULTURAL RIGHTS-OF- WAY/FENCEROW S/HEDGEROW S 12 Ground 687.59 45.X4 ALFALFA 2.4 Ground 10.63 0.71 2.4 Aircraft 13.69 0.91 86 ------- APPLE 3.2 Ground 11.68 0.78 ARTICHOKE 3.2 Ground 32.00 2.13 ASPARGUS 4.8 Ground 36.99 2.47 BANANA, PLANTAIN 2.4 Ground 54.21 3.61 BLACKBERRY, BOYSENBERRY 1.6 Ground 139.70 «UI BLUEBERRY 2.4 Ground 4.65 0.31 CITRUS 3.2 Ground 6.83 0.46 CORN, FIELD 0.8 Ground 7.24 0.48 DEWBERRY 2.4 Ground 19.71 1.31 FILBERT (HAZELNUT) 2.2 Ground 18.90 1.2ft GRAPE 3.2 Ground 9.71 0.65 LOGANBERRY, RASPBERRY (BLACK/RED) 2.4 Ground 5.13 0.34 OLIVE 1.6 Ground 7.17 0.48 PAPAYA 4 Ground 25.21 1.68 PEPPERMINT 2.4 Ground 4.17 0.28 PEACH 3 Ground 10.95 0.73 PEAR 3.2 Ground 11.66 0.78 PECAN 3.2 Ground 27.04 1.80 87 ------- SORGHUM 4.8 Ground 5.87 0.39 SPEARMINT 2.4 Ground 106.70 "Ml WALNUT (ENGLISH/BLACK) 0.4 Ground 25.78 l."72 WHEAT 2.4 Pre-Harvest 18.03 1.20 3 Post- Harvest 12.82 0.85 COTTON 1.6 Ground 8.36 0.56 Aircraft 14.25 0.95 AIRPORTS/ LANDING FIELDS, DRAINAGE SYSTEMS, INDUSTRIAL AREAS (OUTDOOR), SEWAGE DISPOSAL AREAS 12 Ground 2190.00 146.00 BERMUDAGRASS 2.4 Ground 226.00 I5JP GRASSES GROWN FOR SEED 12 Ground 3.76 0.25 IRRIGATION SYSTEMS 1.6 Ground 52.56 3.50 ORNAMENTAL HERBACEOUS PLANTS 12 Ground 36.55 2.44 NON-AGRICULTURAL RIGHTS OF WAY 12 Ground 629.91 41.99 4 Aircraft 626.91 41.^9 88 ------- PAVED AREAS (PRIVATE ROADS/SIDEWALKS) 12 Ground 4911.00 32_7.4<> UNCULTIVATED AG 12 Ground 1130.00 75.33 12 Ground 1169.00 "7"7.')3 UNCULTIVATED NON-AG 12 Ground 284.80 ix.<)y aLOC exceedances (RQ > 1) are bolded and shaded. RQ = use-specific peak EEC/ [Lemna major EC50 = 2.40 ppb]. 5.1.2 Exposures in the Terrestrial Habitat 5.1.2.1 Direct Effects to Terrestrial-phase CRLF As discussed in Section 3.3, potential direct effect determinations to terrestrial-phase CRLFs are based on foliar applications of diuron. Potential direct acute effects to the terrestrial-phase CRLF are derived by considering dose- and dietary-based EECs modeled in T-REX for a small bird (20 g) consuming small invertebrates (Table 3.5) and acute oral and subacute dietary toxicity endpoints for avian species. The Bobwhite Quail exhibited the greatest acute sensitivity to diuron and was therefore selected to serve as a surrogate for the CRLF. Potential direct chronic effects of diuron to the terrestrial-phase CRLF are derived by considering dietary-based exposures modeled in T-REX for a small bird (20g) consuming small invertebrates. Chronic effects are estimated using the lowest available toxicity data for birds. EECs are divided by toxicity values to estimate chronic dietary-based RQs. As mentioned, there are no acceptable data for chronic or dose based acute toxicity of diuron to amphibians or their avian surrogates. Therefore, no RQs were calculated for these values. Linuron, a similar chemical of the same class, has acceptable chronic avian data. The avian reproductive studies for linuron (MRID 42541801, 42541802) using mallard duck and bobwhite quail found reproductive effects at 300 ppm ai with an NOAEL being established at 100 ppm. The endpoints affected for mallard duck reproduction are egg production, adult body weight, feed consumption, viable embryos of eggs set, number of viable embryos, and number of live embryos. For the bobwhite quail the endpoints affected are hatchability and offspring survivability. This suggests that diuron may have chronic effects to avian species, but these risks cannot be quantified 89 ------- in the absence of data. As a result, chronic risks are assumed to avian surrogates for all uses of diuron in California. The most sensitive dose based LD50 was >2000 ppm ai as exhibited by the Mallard Duck (MRID 00160000). It is important to note that although no mortality was observed in this study, ataxia persisted for lid after study concluded. This sublethal effect could potentially result in the animals being more susceptible to predation as a result of being both conspicuous to predators and by having a diminished ability to avoid capture. The calculated acute dietary RQs for the terrestrial phase CRLF exceed the Agency's LOC (0.1) for all uses except sorghum. The RQs in Table 5.5 below are based on the 5-day dietary LC50 of 1730 ppm for bobwhite quail. The acute dietary based RQs exceed the LOC for all uses except Field Corn and Sorghum. The exceedances range from 1.55 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 0.12 (grass for seed, wheat). These results are summarized in Table 5.5. Based on acute dietary LOC exceedances for the listed bird surrogate for amphibians, diuron May Affect the terrestrial-phase of the CRLF directly. Table 5.5 Summary of Acute RQs Used to Estimate Direct Effects to the Terrestrial- phase CRLF Use (Application Rate) l)ielar\ -l>asi-d .Willi- rq! Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non-Agricultural Areas Alfalfa, Peppermint, 0.19 Apple, Grape 0.2'> Citrus, 0.33 Artichoke, Asparagus 0.25 Banana, Plantain 0.54 Bermuda grass, Blackberry, Boysenberry, Spearmint 0.4S Blueberry 0.14 Field Corn 0 ()() Cotton 0.2f. Dewberry 0.2f. Filbert (Hazelnut) 0.IS Grass Seed, Wheat 0.12 Loganberry, Raspberry 0.19 Olive 0.13 Pear, Pecan 0.25 Ornamental Herbaceous plants, Papaya 0.31 Uncultivated Agricultural Areas, Paved Areas 0.94 Peach 0.23 Sorghum 0.03 90 ------- I si' (Application K;ik-) l)kl;ir\ -l>;isi-d .Willi- RQ: Walnul 0.25 * = LOC exceedances (acute RQ > 0.1) are bolded and shaded. 2Based on Bobwhite Quail LC50 =1730 ppm]. 5.1.2.2 Indirect Effects to Terrestrial-Phase CRLF via Reduction in Prey (terrestrial invertebrates, mammals, and frogs) 5.1.2.2.1 Terrestrial Invertebrates In order to assess the risks of diuron to terrestrial invertebrates, which are considered prey of CRLF in terrestrial habitats, the honeybee is used as a surrogate for terrestrial invertebrates. The toxicity value for terrestrial invertebrates is calculated by multiplying the lowest available acute contact LD50 of >145.03 |ig a.i. /bee by 1 bee/0.128g, which is based on the weight of an adult honey bee. The toxicity value for terrestrial invertebrates is calculated to be >1133.05 ppm, but only 2.77% mortality was observed at the highest levels tested. Therefore, this test is not definitive and acute RQs were not calculated. However, because the terrestrial EEC's exceed the highest levels tested for several uses, it is determined that diuron May Affect the CRLF indirectly via reduction in terrestrial invertebrate prey items. 5.1.2.2.2 Mammals Risks associated with ingestion of small mammals by large terrestrial-phase CRLFs are derived for dietary-based and dose-based exposures modeled in T-REX for a small mammal (15g) consuming short grass. Acute and chronic effects are estimated using the most sensitive mammalian toxicity data. EECs are divided by the toxicity value to estimate acute and chronic dose-based RQs as well as chronic dietary-based RQs. Chronic dose based RQs exceed the Agency's LOC (1.0) for most uses ( Table 5.6). The dose based chronic RQs that exceed the LOC range from 8.29 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 1.31 (walnut). The dietary based chronic RQs do not exceed the LOC for any uses. The RQs that exceed the acute dose based LOC range from 0.41 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 0.13 (Bermuda grass, Blackberry, Boysenberry, Spearmint). Based on acute and chronic LOC exceedances on small mammal prey items, diuron May Affect the CRLF indirectly via reduction in small mammal prey items. 91 ------- Table 5.6 Summary of Acute and Chronic RQs* Used to Estimate Indirect Effects to the Terrestrial-phase CRLF via Direct Effects on Small Mammals as Dietary Food Items l si- ( linink KQ1 .Willi- KQ l)k'l;ir\-l>;iM-(l l)iiM'-l>;iM'(l (lironk KQ (liroiik KQ .Willi' KQ Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non-Agricultural Areas N.2'> 0.96 0.41 Alfalfa, Peppermint 1.00 0.12 0.05 Apple, Grape 1.5ft 0.18 0.08 Artichoke, Asparagus 1.33 0.15 0.07 Banana, Plantain 2.87 0.33 0.14 Bermuda grass, Blackberry, Boysenberry, Spearmint 2.57 0.30 0.13 Blueberry 0.74 0.09 0.04 Citrus 1.74 0.20 0.09 Field Corn 0.33 0.04 0.02 Cotton 1.40 0.16 0.07 Dewberry 1.40 0.16 0.07 Filbert (Hazelnut) 0.96 0.11 0.05 Grass Seed, Wheat 0.67 0.08 0.03 Loganberry, Raspberry 1.00 0.12 0.05 Olive 0.69 0.08 0.03 Ornamental Herbaceous Plants, Papaya I.ft7 0.19 0.08 Uncultivated Agricultural Areas, Paved Areas 5.00 0.58 0.25 Peach 1.25 0.14 0.06 Pear, Pecan 1.33 0.15 0.07 Sorghum 0.17 0.02 0.01 Walnut 1.31 0.15 0.07 * = LOC exceedances (acute RQ >0.1 and chronic RQ > 1) arc bolded and shaded. 1 Based on dose-based EEC and diuron rat NOAEC = 250.00 mg/kg-bw. 2 Based on dose-based EEC and diuron rat acute oral LD50 = 5000.00 mg/kg-bw. 5.1.2.2.3 Frogs An additional prey item of the adult terrestrial-phase CRLF is other species of frogs. In order to assess risks to these organisms, dietary-based and dose-based exposures modeled 92 ------- in T-REX for a small bird (20g) consuming small invertebrates are used. See Section 5.1.2.1 for results. The chronic LOC is exceeded for all uses of diuron and the acute LOCs are exceeded for all uses except sorghum. Based on acute and chronic LOC exceedances on the prey item of small frogs, diuron May Affect the CRLF indirectly via reduction in small frog prey items. 5.1.2.3 Indirect Effects to CRLF via Reduction in Terrestrial Plant Community (Riparian and Upland Habitat) Potential indirect effects to the CRLF resulting from direct effects on riparian and upland vegetation are assessed using RQs from terrestrial plant seedling emergence and vegetative vigor EC25 data as a screen. Diuron is a systemic herbicide. The RQs for non-target terrestrial monocot and dicot plants inhabiting semi-aquatic and upland dry areas exceed the Agency's LOC (1.0) for all uses except for monocot plants exposed to sorghum applications (Table 5.7). These exceedances range from 300.00 (semi-aquatic plants exposed to non-agricultural aerial applications) to 0.19 (dicot plants inhabiting dry areas exposed to sorghum applications). Several diuron uses result in LOC exceedances from spray drift. These exceedances range from 12.00 (dicot plants exposed to field corn applications) to 1.13 (monocot plants exposed to cotton applications). Based on LOC exceedances on the non-target terrestrial plants at all use sites, diuron May Affect the terrestrial phase CRLF indirectly via effects to riparian and upland habitat. Table 5.7 RQs*1^ for Non-Target Plants Inhabiting Dry and Semi-Aquatic Areas Exposed to Diuron via Runoff and Drift Use Application rate (lbs a.i./A) Application method Drift Value (%) Group Sprav drift RQ Dry area RQ Semi- aquatic area RQ Agricultural Rights-of- Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non- Agricultural Areas 12.0 Foliar/Ground 1 Monocot 3.M 25.45 5.71 Dicot 4. SO 33.60 r.o.oo Alfalfa, Peppermint, Spearmint, Bermudagrass, Blackberry, Boysenberry, Loganberry, Raspberry, Dewberry 2.4 Foliar/Ground 1 Monocot 0.73 5.09 1.14 Dicot 0.96 h.72 12.00 93 ------- Use Application rate (lbs a.i./A) Application method Drift Value (%) Group Sprav drift RQ Dry area RQ Semi- aquatic area RQ Apple, Citrus, Pear, Grape, Artichoke, Asparagus, Pecan 3.2 Foliar/Ground 1 Monocot 0.97 h.l') 1.52 Dicot I.2S X.'H, 16.00 Asparagus (Aerial) 3.2 Foliar/Ground 1 Monocot 2.2f. S.OS 7.62 Dicot 2.'W 10.67 S0.00 Banana, Plantain 4.8 Foliar/Ground 1 Monocot 1.45 10. IS 2.2') Dicot l.')2 13.44 24.00 Blueberry, Grass Seed, Olive, Pear, Wheat 1.6 Foliar/Ground 1 Monocot 0.48 3.3') 0.76 Dicot 0.64 4.4S S.00 Field Corn 0.8 Foliar/Ground 1 Monocot 0.24 1.70 0.38 Dicot 12.00 S4.00 4.00 Cotton 3 Aerial 5 Monocot 1.13 4.04 3.SI Dicot 1.4') 5.33 40.00 Cotton 3 Ground 1 Monocot 0.48 3.3') 0.76 Dicot 24.0 K.N.O S.00 Filbert (Hazelnut) 2.2 Foliar/Ground 1 Monocot 0.67 4.67 1.05 Dicot 0.88 6.16 11.00 Non-agricultural 12 Aerial 5 Monocot S.4S 30.30 28.57 Dicot 11.20 40.00 300.00 Ornamental Herbaceous Plants, Papaya 4 Foliar/Ground 1 Monocot 1.21 S.4S l.'JO Dicot l.r.o 11.20 20.00 Uncultivated Agricultural Areas, Paved Areas 12 Foliar/Ground 1 Monocot 3.(> 4 25.45 5.71 Dicot 4. SO 33.60 60.00 Peach, Walnut 3 Foliar/Ground 1 Monocot 0.91 r,.3f. 1.43 Dicot 1.20 S.40 15.00 Sorghum 0.4 Foliar/Ground 1 Monocol 0.12 0.85 0.19 Dicot 0.16 1.12 2.00 * LOC Exceedances (RQ > 1.0) are bolded and shaded. ^ Spray drift RQs consider only spray drift alone, dry area and semi-aquatic RQs consider spray drift as well as runoff. 5.1.3 Primary Constituent Elements of Designated Critical Habitat 5.1.3.1 Aquatic-Phase (Aquatic Breeding Habitat and Aquatic Non- Breeding Habitat) Three of the four assessment endpoints for the aquatic-phase primary constituent elements (PCEs) of designated critical habitat for the CRLF are related to potential effects to aquatic and/or terrestrial plants: 94 ------- • Alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond: aquatic habitat (including riparian vegetation) provides for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. • Alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food source. • Reduction and/or modification of aquatic-based food sources for pre-metamorphs (e.g., algae). Based on the risk estimation for potential effects to vascular and non-vascular aquatic and terrestrial plants provided in Sections 5.1.1.2, 5.1.1.3, and 5.1.2.3, diuron may result in Habitat Modification of aquatic-phase PCEs of designated habitat related to effects on aquatic and/or terrestrial plants. • Aquatic non-vascular plants used as food source and habitat for CRLF may be potentially affected from all diuron uses. • Reduction of aquatic based food sources may occur from most use sites. • Due to aquatic vascular and terrestrial plant communities being reduced from most use sites, there is potential for alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond. • Due to aquatic vascular and terrestrial plant communities being reduced from most use sites, there is potential for alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food. The remaining aquatic-phase PCE is "alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source." To assess the impact of diuron on this PCE, acute and chronic freshwater fish and invertebrate toxicity endpoints, as well endpoints for aquatic non-vascular plants are used as measures of effects. RQs for these endpoints were calculated in Sections 5.1.1.1 and 5.1.1.2. Based on acute and chronic LOC exceedances for freshwater fish and aquatic invertebrates, diuron may result in Habitat Modification of aquatic-phase PCEs related to effects of alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source. 5.1.3.2 Terrestrial-Phase (Upland Habitat and Dispersal Habitat) Two of the four assessment endpoints for the terrestrial-phase PCEs of designated critical habitat for the CRLF are related to potential effects to terrestrial plants: • Elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs: Upland areas within 200 ft of the edge of the riparian vegetation or dripline surrounding aquatic and riparian habitat that are comprised 95 ------- of grasslands, woodlands, and/or wetland/riparian plant species that provides the CRLF shelter, forage, and predator avoidance • Elimination and/or disturbance of dispersal habitat: Upland or riparian dispersal habitat within designated units and between occupied locations within 0.7 mi of each other that allow for movement between sites including both natural and altered sites which do not contain barriers to dispersal The risk estimation for terrestrial-phase PCEs of designated habitat related to potential effects on terrestrial plants is provided in Section 5.1.2.3. These results will inform the effects determination for modification of designated critical habitat for the CRLF. The third terrestrial-phase PCE is "reduction and/or modification of food sources for terrestrial phase juveniles and adults." To assess the impact of diuron on this PCE, acute and chronic toxicity endpoints for birds, mammals, and terrestrial invertebrates are used as measures of effects. RQs for these endpoints were calculated in Section 5.1.2.2. Based on acute and chronic LOC exceedances for CRLF prey items of small mammals, terrestrial invertebrates and other frogs, diuron may result in Habitat Modification of the first three terrestrial phase PCEs. The fourth terrestrial-phase PC is based on alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. Direct acute and chronic RQs for terrestrial-phase CRLFs are presented in Section 5.2.1.2. Due to acute and chronic LOC exceedances at all use sites to terrestrial-phase CRLFs, diuron may result in Habitat Modification of the fourth terrestrial phase PCE. 5.2 Risk Description The risk description synthesizes an overall conclusion regarding the likelihood of adverse impacts leading to an effects determination (i.e., "no effect," "may affect, but not likely to adversely affect," or "likely to adversely affect") for the CRLF and its designated critical habitat. If the RQs presented in the Risk Estimation (Section 5.1) show no direct or indirect effects for the CRLF, and no modification to PCEs of the CRLF's designated critical habitat, a "no effect" determination is made, based on diuron's use within the action area. However, if direct or indirect effects LOCs are exceeded, or if effects may modify the PCEs of the CRLF's critical habitat, the Agency concludes a preliminary "may affect" determination for the FIFRA regulatory action regarding diuron. Based on direct and indirect LOC exceedances for CRLF, the Agency concludes a preliminary May Affect determination for the CRLF and critical habitat. A summary of the results of the risk estimation is provided in Table 5.8 for direct and indirect effects to the CRLF and in Table 5.9 for the PCEs of designated critical habitat for the CRLF. 96 ------- Table 5.8 Risk Estimation Summary for Diuron - Direct and Indirect Effects to CRLF Assessment Endpoint LOC Excccdanccs (Y/N) Description of Results of Risk Estimation Aquatic Phase (eggs, larvae, tadpoles, juveniles, and adults) Direct Effects Survival, growth, and reproduction of CRLF individuals via direct effects on aquatic phases YES The aquatic phase amphibian acute LOCs for Listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 12.28 (paved areas) to 0.06 (papaya and walnut). Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 131.85 (paved areas) to 1.26 (banana, plantain). Indirect Effects Survival, growth, and reproduction of CRLF individuals via effects to food supply {i.e., freshwater invertebrates, non-vascular plants) YES LOCs for non-vascular plants are exceeded for all uses. The RQs range from 2046.25 (paved areas) to 1.57 (grasses grown for seed). Aquatic invertebrates acute a LOC are exceeded. The acute RQs that exceed the LOC range from 30.69 (paved areas) to 0.05 (cotton aerial application). Chronic LOCs are exceeded for several uses. RQs that exceed the Agency's LOC range from 21.21or paved areas to 1.02 for bermudagrass. Indirect Effects Survival, growth, and reproduction of CRLF individuals via effects on habitat, cover, and/or primary productivity (i.e., aquatic plant community) YES RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 460.46 (paved areas) to 1.30 (hazelnut). Indirect Effects Survival, growth, and reproduction of CRLF individuals via effects to riparian vegetation, required to maintain acceptable water quality and habitat in ponds and streams comprising the species' current range. YES RQs for non-target terrestrial monocot and dicot plants inhabiting semi- aquatic and upland dry areas exceed the Agency's LOC for all uses except for monocot plants exposed to sorghum applications. RQs range from 300.00 (semi-aquatic plants exposed to non-agricultural aerial applications) to 0.19 (dicot plants inhabiting dry areas exposed to sorghum applications). Terrestrial Phase (Juveniles and adults) Direct Effects Survival, growth, and reproduction of CRLF individuals via direct effects on terrestrial phase adults and juveniles YES The acute dietary based RQs that exceed the LOC range from 1.55 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 0.12 (grass for seed, wheat). These results are summarized in Table 5.5. Indirect Effects Survival, growth, and reproduction of CRLF individuals via effects on prey (i.e., terrestrial invertebrates, small terrestrial mammals and terrestrial phase amphibians) YES Chronic dose based RQs exceed the Agency's LOC (1.0) for most uses ( Table 5.6). The dose based chronic RQs that exceed the LOC range from 8.29 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 1.31 (walnut). The dietary based chronic RQs do not exceed the LOC for any uses. The RQs that exceed the acute dose based LOC range from 0.41 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.) to 0.13 (Bermuda grass, Blackberry, Boysenberry, Spearmint). Indirect Effects Survival, growth, and reproduction of CRLF individuals via effects on habitat (i.e., riparian vegetation) YES The RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses of diuron in California. These range from 373.33 (paved areas) to 1.01 (alfalfa aerial). The RQs for non-target terrestrial monocot and dicot plants inhabiting semi-aquatic and upland dry areas exceed the Agency's LOC (1.0) for all 97 ------- uses except for monocot plants exposed to sorghum applications (Table 5.7). These exceedances range from 300.00 (semi-aquatic plants exposed to non-agricultural aerial applications) to 0.19 (dicot plants inhabiting dry areas exposed to sorghum applications). Several diuron uses result in plant LOC exceedances from spray drift. These exceedances range from 12.00 (dicot plants exposed to field corn applications) to 1.13 (monocot plants exposed to cotton applications). Table 5.9 Risk Estimation Summary for Diuron - PCEs of Designated Critical Habitat for the CRLF Assessment Endpoint LOC Exceedances (Y/N) Description of Results of Risk Estimation Aquatic Phase PCEs (Aquatic Breeding Habitat and Aquatic Non-Breeding Habitat) Alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond: aquatic habitat (including riparian vegetation) provides for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. YES LOCs are exceeded for terrestrial riparian plants and for aquatic vascular plants from exposure to diuron from runoff or spray drift. Alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food source. YES LOCs are exceeded for terrestrial riparian plants and for aquatic plants from exposure to diuron from runoff or spray drift. Alteration of riparian and vascular plants may result in alteration of temperature, turbidity, and oxygen content. Alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source. YES LOC is exceeded for indirect effects on terrestrial and aquatic-phase CRLF from most diuron applications. Reduction and/or modification of aquatic-based food sources for pre-metamorphs (e.g., algae) YES LOCs for non-vascular plants are exceeded for all uses. Terrestrial Phase PCEs (Upland Habitat and Dispersal Habitat) Elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs: Upland areas within 200 ft of the edge of the riparian vegetation or dripline surrounding aquatic and riparian habitat that are comprised of grasslands, woodlands, and/or wetland/riparian plant species that provides the CRLF shelter, forage, and predator avoidance YES The AgDRIFT model was used to evaluate potential distances beyond which exposures would be expected to be below LOC. However, due to the limitations imposed by the Tier 1 ground analysis (allows users to evaluate off-site deposition and exposure out to 1,000 ft downwind from the location of the application), the exact buffer needed for exposures to be below the LOC is uncertain. The output from AgDRIFT indicated that the buffer zone required would be greater than 1,000 feet. Since the model is restricted to accurately discerning a buffer within 1,000 feet of the application, the exact distance needed for a buffer to protect non-listed and listed plants is unknown. Flowever, as seen in Table 3.7, the calculated risk quotient (RQ) is significantly larger than the level of concern (LOC) for non-listed and listed plant species (LOC =1). 98 ------- Assessment Endpoint LOC Excccdanccs (Y/N) Description of Results of Risk Estimation This comparison provides awareness to approximately how much greater than 1,000 feet the buffer needs to be. Elimination and/or disturbance of dispersal habitat: Upland or riparian dispersal habitat within designated units and between occupied locations within 0.7 mi of each other that allow for movement between sites including both natural and altered sites which do not contain barriers to dispersal YES Effects are expected to non-target terrestrial plants over 1000 ft from use site from ground application. Reduction and/or modification of food sources for terrestrial phase juveniles and adults YES LOC is exceeded for indirect effects on terrestrial and aquatic-phase CRLF from diuron application Alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. YES LOC is exceeded for direct effects on terrestrial and aquatic-phase CRLF from diuron applications. Following a preliminary "may affect" or "habitat modification" determination, additional information is considered to refine the potential for exposure at the predicted levels based on the life history characteristics {i.e., habitat range, feeding preferences, etc.) of the CRLF. Based on the best available information, the Agency uses the refined evaluation to distinguish those actions that "may affect, but are not likely to adversely affect" from those actions that are "likely to adversely affect" the CRLF and its designated critical habitat. The criteria used to make determinations that the effects of an action are "not likely to adversely affect" the CRLF and its designated critical habitat include the following: • Significance of Effect: Insignificant effects are those that cannot be meaningfully measured, detected, or evaluated in the context of a level of effect where "take" occurs for even a single individual. "Take" in this context means to harass or harm, defined as the following: ¦ Harm includes significant habitat modification or degradation that results in death or injury to listed species by significantly impairing behavioral patterns such as breeding, feeding, or sheltering. ¦ Harass is defined as actions that create the likelihood of injury to listed species to such an extent as to significantly disrupt normal behavior patterns which include, but are not limited to, breeding, feeding, or sheltering. • Likelihood of the Effect Occurring: Discountable effects are those that are extremely unlikely to occur. • Adverse Nature of Effect: Effects that are wholly beneficial without any adverse effects are not considered adverse. A description of the risk and effects determination for each of the established assessment endpoints for the CRLF and its designated critical habitat is provided in Sections 5.2.1 through 5.2.3. 99 ------- 5.2.1 Direct Effects 5.2.1.1 Aquatic-Phase CRLF The aquatic-phase considers life stages of the frog that are obligatory aquatic organisms, including eggs and larvae. It also considers submerged terrestrial-phase juveniles and adults, which spend a portion of their time in water bodies that may receive runoff and spray drift containing diuron. Diuron is considered "highly toxic" to the freshwater fish, which are surrogates for the aquatic phase CRLF. The aquatic animal acute LOCs for Listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 12.28 (paved areas) to 0.06 (papaya). The probability of an individual effect to the aquatic-phase CRLF is based on the slope of the dose response curve for the acute endpoint used to derive the RQ. The probability, based on the default slope of 4.5 is one in 4.18E+08. Because the upper and lower 95% confidence interval of the slope is not known, a plausible range of slopes of 2 to 9 were used to obtain reasonable upper and lower bounds. The resulting probability of an individual effect to the aquatic-phase CRLF range between one in 215.83 and one in 1.75E+31. Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 131.85 (paved areas) to 1.26 (banana, plantain). The aquatic chronic animal LOC for listed species (1.0) is exceeded for most uses of diuron in California. The USGS collected 422 positive ground water samples containing Diuron. The three river basins looked at were Sacramento, San Joaquin-Tulare, and Santa Ana. The maximum concentration observed was 1.8 ppb in the San Joaquin-Tulare water basins in Kings County. Monitoring data from CDPR indicate that diuron concentrations range from non- detectable to 160 ppb. The majority of the large concentrations of Diuron came from San Joaquin County, and occurred between the end of December and the beginning of January. The year 2005, for San Joaquin County, yielded the greatest concentrations overall. Moreover, the CDPR monitoring data showed that eleven samples (1.56% of the 705 samples), ranging from 31 ppb to 160 ppb, detected levels of diuron that would result in chronic direct effects to the CRLF. Likewise, four samples (0.57%), ranging from 9.5 ppb to 15 ppb, detected diuron at levels that would result in chronic indirect effects to the CRLF. These detects occurred during the months of December, January, and February in years 2005 and 2006, and in March for the 2006 year. The San Joaquin County had 5 detects (3.38% of samples) resulting in chronic direct effects and 1 detect (0.68% of samples) resulting in chronic indirect effects, Tulare County had 1 detect (2.22% of samples) resulting in a chronic direct effect, and Stanislaus County had 5 detects (6.94% of samples) resulting in chronic direct effects and 2 detects (2.78% of samples) resulting 100 ------- in chronic indirect effects. Consequently, almost 10% of the samples taken from Stanislaus County, just over 4% from San Joaquin County, a little over 2% from Tulare County contained detects at levels that would cause chronic or indirect effects to the CRLF. Seven reported incidents resulted in fish kills. Five of these fish kills resulted from direct application of diuron to water bodies which is not allowed in California. However, three incidents were from drift or runoff of diuron from applications made adjacent to water bodies. This demonstrates that diuron may pose a threat to aquatic organisms through labeled uses (Appendix J). It is also important to note that the absence of additional documented incidents does not necessarily mean that such incidents did not occur. Mortality incidents must be seen, reported, investigated, and submitted to the Agency in order to be recorded in the incident database. Incidents may not be noticed because the carcasses decayed, were removed by scavengers, or were in out-of-the-way or hard-to-see locations. Due to the voluntary nature of incident reporting, an incident may not be reported to appropriate authorities capable of investigating it. Based on these exceedances to the aquatic-phase CRLF, reported diuron incidents resulting in fish kills and its monitored presence in surface water, a "May Affect and Likely to Adversely Affect (LAA)" determination is made for diuron use in California. 5.2.1.2 Terrestrial-Phase CRLF The RQs representing acute dietary-based exposures exceed the Agency's LOC for all uses except sorghum (Section 5.1.2.1.). These RQs were derived using the T-REX model, which estimates exposures that are specific to food intake equations for birds. RQs generated for birds are used as surrogates to represent RQs for the terrestrial-phase CRLF. Based on these exceedances to the terrestrial-phase CRLF, a "May Affect" determination was made. The T-HERPS model was therefore employed as a refinement tool to explore amphibian- specific food intake on potential exposures to the terrestrial phase CRLF. The T-HERPS model incorporates the same inputs as T-REX with equations adjusted for poikilotherm food intake. The EECs generated by T-HERPS are found in Table 5.10. An example output from T-HERPS is available in Appendix K. 101 ------- Table 5.10. Upper-bound Kenega Nomogram T-HERPS EECs (mg/kg-diet) for Dietary-based Exposures of the CRLF and its Prey to Diuron1. Scenario Small Insects Large Insects Small Herbivore Mammals Small Insectivore Mammals Small Terrestrial Phase Amphibians Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non- Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non-Agricultural Areas 2113.70 234.86 2476.10 154.76 73.37 Alfalfa, Peppermint, 324.00 36.00 379.55 23.72 11.25 Apple, Grape 504.68 56.08 591.21 36.95 17.52 Citrus Artichoke, Asparagus 432.00 48.00 506.07 31.36 15.00 Banana, Plantain 930.06 103.34 1089.52 68.10 32.28 Bermuda grass, Blackberry, Boysenberry, Spearmint 951.44 105.72 114.57 69.66 33.03 Blueberry 239.50 23.61 280.57 17.54 8.31 Field Corn 108.00 12.00 126.52 7.91 3.75 Cotton 452.53 50.28 530.11 33.13 15.71 Dewberry 452.83 50.31 530.47 33.15 15.72 Filbert (Hazelnut) 312.23 34.69 365.76 22.86 10.84 Grass Seed, Wheat 216.00 24.00 253.03 15.81 7.50 Loganberry, Raspberry 324.00 36.00 379.55 23.72 11.25 Olive 223.75 24.86 262.12 16.38 7.77 Pear, Pecan 432.00 48.00 506.07 31.63 15.00 Ornamental Herbaceous Plants, Papaya 540.00 60.00 632.59 39.54 18.74 Uncultivated Agricultural Areas, Paved Areas 1620.00 180.00 1897.76 118.61 56.23 Peach 405.00 45.00 474.44 29.65 14.06 Sorghum 54.00 6.00 63.26 3.95 1.87 Walnut 425.76 47.31 498.76 31.17 14.78 'weights of small herbivore and insectivore mammals are 35g 102 ------- Acute exposures Refined acute dietary-based RQs for CRLFs consuming large insects and small herbivore mammals exceed the acute listed species LOC (0.1) for all uses of diuron except Sorghum and Field Corn. The acute dietary-based RQs for CRLFs consuming large insects exceed the acute listed species LOC for Agricultural Rights-of-Way, Fencerows etc. No acute dietary based LOCs were exceeded for CLRF consuming small insectivore mammals and small terrestrial phase amphibians for any diuron use. Results are presented in Table 5.11. Table 5.11. Revised Acute Dietary-based RQs for CRLF consuming different food items (RQs calculated using T-HERPS)*. Scenario Small 111 sec Is 1 :l I'm' 111 sills Small lli'i'lmiiiv Maiiiinals Small Insectivore Mammals Small Terrestrial Phase Amphibians Agricultural Rights-of- Way, Fencerows, Hedgerows, Airports, landing Fields, Drainage Systems, Outdoor Industrial Areas, Irrigation Systems, Non-Agricultural Rights of Way, Sewage Disposal Areas, Uncultivated Non- Agricultural Areas 1.22 0.14 1.43 0.09 0.04 Alfalfa, Peppermint, O.I') 0.02 0.22 0.01 0.01 Apple, Grape 0.2') 0.03 0.34 0.02 0.01 Citrus 0.33 0.04 0.3S 0.02 0.01 Artichoke, Asparagus 0.25 0.03 0.2') 0.02 0.01 Banana, Plantain 0.54 0.06 0.ft3 0.04 0.02 Bermuda grass, Blackberry, Boysenberry, Spearmint 0.55 0.06 11.64 0.04 0.02 Blueberry 0.14 0.02 0.1 ft 0.01 <0.01 Field Corn 0.06 0.01 () ()7 <0.01 <0.01 Cotton 0.2ft 0.03 0.31 0.02 <0.01 Dewberry 0.2ft 0.03 0.31 0.02 0.01 Filbert (Hazelnut) O.IS 0.02 0.21 0.01 0.01 Grass Seed, Wheat 0.12 0.01 0.15 0.01 <0.01 103 ------- Loganberry, Raspberry 0.1') 0.02 0.22 0.01 0.01 Olive 0.13 0.01 0.15 0.01 <0.01 Ornamental Herbaceous plants, Papaya 0.31 0.03 0.37 0.02 0.01 Uncultivated Agricultural Areas, Paved Areas O.'M 0.10 1.10 0.07 0.03 Peach 0.23 0.03 0.27 0.02 0.01 Pear, Pecan 0.25 0.03 0.2'> 0.02 0.01 Sorghum 0.03 <0.01 0.04 <0.01 <0.01 Walnut 0.25 0.03 0.2'> 0.02 0.01 *RQs exceeding the Listed LOC (0.10) are bolded and shaded Based on the dietary probit dose-response slope of 7.22 the chance of individual mortality for which RQs exceed the Listed LOC range from approximately one in 6.75E+10 at an RQ of 0.12 (Grass Seed and Wheat uses) to approximately one in one at an RQ of 1.22 (Agricultural Rights-of-Way, etc.). Therefore, based on these refined dietary based acute risk quotients and their exceedances of the Agency's LOC a Likely to Adversely Affect (LAA) determination is made for diuron use in California. 5.2.2 Indirect Effects (via Reductions in Prey Base) 5.2.2.1 Algae (non-vascular plants) Indirect effects of diuron to the aquatic-phase CRLF (tadpoles) via reduction in non- vascular aquatic plants in its diet are based on peak EECs from the standard pond and the lowest acute toxicity value for aquatic non-vascular plants. The Agency's LOC (1.0) is exceeded for all current uses of diuron in California. The RQs range from 2331.67 for Paved Areas to 1.82 for Sorghum (Section 5.1.1.2). The fate characteristics indicate that diuron is expected to be persistent in aquatic environments with EECs after 60 days being above the Agency's LOC for non-vascular aquatic plants, a primary food source for aquatic-phase CRLF. The application of diuron in California is anticipated to be in late winter and early spring. The timing of the application would coincide with reproduction of CRLF in aquatic environments as well as for the tadpoles to feed on non-vascular aquatic plants. Because of non-vascular LOC exceedances from registered uses of diuron, its monitored presence in surface water and verified non-target incidents resulting from diuron use, based on multiple lines of evidence, the Agency concludes that there is a potential indirect impact to the aquatic-phase of the CRLF from reduction of food items (algae) and therefore diuron is Likely to Adversely Affect (LAA) the CRLF. 104 ------- 5.2.2.2 Aquatic Invertebrates As discussed in Section 2.5.3, the diet of CRLF also includes aquatic invertebrates. The potential for diuron to elicit indirect effects to the CRLF via effects on freshwater invertebrate food items is dependent on several factors including: (1) the potential magnitude of effect on freshwater invertebrate individuals and populations; and (2) the number of prey species potentially affected relative to the expected number of species needed to maintain the dietary needs of the CRLF. Together, these data provide a basis to evaluate whether the number of individuals within a prey species is likely to be reduced such that it may indirectly affect the CRLF. The main food source for juvenile aquatic- and terrestrial-phase CRLFs is thought to be aquatic and terrestrial invertebrates found along the shoreline and on the water surface. The application of diuron in California is anticipated to be in late winter and early spring. The timing of the application would coincide with juvenile aquatic- and terrestrial-phase CRLFs that would be feeding on aquatic and terrestrial invertebrates. Indirect acute effects to the aquatic-phase CRLF via effects to prey (invertebrates) in aquatic habitats are based on peak EECs in the standard pond and the lowest acute toxicity value for freshwater invertebrates. For chronic risks, 21-day EECs and the lowest chronic toxicity value for invertebrates are used to derive RQs. The Agency's acute Listed LOC (0.05) is exceeded for most uses of diuron in California. The acute RQs that exceed the LOC range from 30.69 (Paved Areas) to 0.06 (grape). The chronic RQs exceed the Agency's LOC for several used. The chronic values that exceeded range from 21.21 (paved areas) to 1.02 (bermudagrass). A summary of the acute and chronic RQ values for exposure to aquatic invertebrates (as prey items of aquatic-phase CRLFs) is provided in Table 5.3. Based on acute Listed LOC exceedances for chronic and acute aquatic invertebrates from most use sites, diuron May Affect the CRLF indirectly via reduction in freshwater invertebrate prey items. Based on the aquatic invertebrate RQs and a probit dose-response default slope of 4.5 the estimated reduction in aquatic invertebrate populations for which RQs exceed the Listed LOC range from <0.01% to approximately 100%. To further investigate the estimated reduction in aquatic invertebrate populations, a plausible range of slopes of 2 to 9 were selected to investigate the upper and lower bounds of this estimate. Using a slope of 2 led to a range of possible reduction in populations from less than 0.10% to approximately 100%). Employing a slope of 7 led to a range of possible reduction in aquatic invertebrate populations from less than 1.0% to approximately 100%. Because aquatic invertebrate populations are expected to be impacted from registered uses of diuron in California, and because diuron's presence has been observed in monitored surface water in California, the Agency concludes that there is a potential indirect impact to the aquatic-phase of the CRLF from a reduction of aquatic invertebrate food items and therefore diuron is Likely to Adversely Affect (LAA) the CRLF. 105 ------- 5.2.2.3 Fish and Aquatic-phase Frogs As discussed in Section 2.5.3, the diet of CRLF also includes small fish and other aquatic-phase frogs. Direct effects to the aquatic-phase CRLF are based on peak EECs in the standard pond and the lowest acute toxicity value for freshwater fish. In order to assess direct chronic risks to the CRLF, 60-day EECs and the lowest chronic toxicity value for freshwater fish are used. The RQs for diuron uses results in acute and chronic exceedances of the Agency's LOC for freshwater fish which are surrogates for the aquatic phase for amphibians. The aquatic phase amphibian acute LOCs for Listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 13.99 (paved areas) to 0.053 (papaya). Based on these RQs and a probit dose-response default slope of 4.5 the estimated aquatic vertebrate prey item population reduction for which RQs exceed the Listed LOC range from approximately 10.0% to approximately 100%. To further investigate the estimated aquatic vertebrate prey item population reduction, a plausible range of slopes of 2 to 9 were selected to investigate the upper and lower bounds of this estimate. Using a slope of 2 led to a range of estimated population reduction of approximately 30.0% to approximately 100%. Employing a slope of 9 led to a range of possible population reduction of approximately 3.0% to approximately 100%. The aquatic phase amphibian acute LOCs for listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 12.28 (paved areas) to 0.06 (papaya and walnut). Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 131.85 (paved areas) to 1.26 (banana, plantain). The application of diuron in California is anticipated to be in late winter and early spring. The timing of the application would coincide with juvenile aquatic- and terrestrial-phase CRLFs that would be feeding on small fish, small frogs or tadpoles. Because fish and aquatic phase amphibian populations are expected to be impacted from registered uses of diuron in California, and because diuron's presence has been observed in monitored surface water in California, the Agency concludes that there is a potential indirect impact to the aquatic-phase of the CRLF from a reduction of fish and aquatic phase amphibian food items and therefore diuron is Likely to Adversely Affect (LAA) the CRLF. 5.2.2.4 Terrestrial Invertebrates When the terrestrial-phase CRLF reaches juvenile and adult stages, its diet is mainly composed of terrestrial invertebrates. Terrestrial invertebrate toxicity data are used to assess potential indirect effects of diuron to the terrestrial-phase CRLF. Effects to terrestrial invertebrates resulting from exposure to diuron may also indirectly affect the CRLF via reduction in available food. Because the estimated EEC's exceed the highest level tested for three uses for diuron, a preliminary "May Affect" determination was made. However, the calculated EEC's for 106 ------- Agricultural Rights-of-Way etc (2688.80 ppm) was only roughly twice the highest level tested (>1305) at which only a 2.7% mortality was observed. Therefore, it is reasonable to assume that the effects to terrestrial invertebrate populations will be negligible for these uses of diuron in California. Therefore, based on the weight-of-evidence, the Agency concludes that there is a negligible potential impact to terrestrial invertebrates that the CRLF consumes, and therefore diuron May Affect but is Not Likely to Adversely Affect (NLAA) the CRLF. 5.2.2.5 Mammals Life history data for terrestrial-phase CRLFs indicate that large adult frogs consume terrestrial vertebrates, including mice. Small mammals can make up to 50% of the CRLF food intake. Acute dose based RQs exceed the Agency's LOC (1.0) for most uses. The dose based RQs that exceed the LOC range from 0.41 (Agricultural Areas, Rights-of- Way, fencelines etc.) to 0.13 (Bermudagrass, Blackberry, Boysenberry, Spearmint). The dose based chronic RQs that exceed the LOC range from 8.29 (Agricultural Areas, Rights-of-Way, fencelines etc.) to 1.33 (Artichoke, Asparagus, Pear, Pecan).. The dietary based chronic RQs do not exceed the LOC for any uses. Based on the mammalian dose based acute RQs and a probit dose-response default slope of 4.5 the estimated mammalian prey item population reduction for which RQs exceed the Listed LOC range from <0.01% to approximately 4.00%. To further investigate the estimated reduction in these populations, a plausible range of slopes of 2 to 9 were selected to investigate the upper and lower bounds of this estimate. Employing a slope of 9 led to an estimated population reduction of <0.01% for all RQs. Using a slope of 2 led to a range of possible population reduction from approximately 4.00% to approximately 22.00%. Because mammalian populations are expected to be impacted from registered uses of diuron in California, the Agency concludes that there is a potential indirect impact to the terrestrial phase of the CRLF from a reduction of mammalian food items and therefore diuron is Likely to Adversely Affect (LAA) the CRLF. 5.2.2.6 Terrestrial-phase Amphibians Terrestrial-phase adult CRLFs also consume frogs. RQ values representing direct exposures of diuron to terrestrial-phase CRLFs are used to represent exposures of diuron to frogs in terrestrial habitats. The T-HERPS model was therefore employed as a refinement tool to explore amphibian-specific food intake on potential exposures to terrestrial phase amphibian food items for the CRLF. The T-HERPS model incorporates the same inputs as T-REX with equations adjusted for poikilotherm food intake. Acute exposures Refined acute dietary-based RQs for terrestrial phase amphibians consuming small insects and small herbivore mammals exceed the acute listed species LOC (0.1) for all uses of diuron except Sorghum and Field Corn. The acute dietary-based RQ for 107 ------- terrestrial phase amphibians consuming large insects exceeds the acute listed species LOC for Agricultural Rights-of-Way, Fencerows etc. Based on these refined dietary based risk quotients, terrestrial phase amphibian populations are expected to be impacted from registered uses of diuron in California. Therefore the Agency concludes that there is a potential indirect impact to the terrestrial phase of the CRLF from a reduction of terrestrial phase amphibian food items and therefore diuron is Likely to Adversely Affect (LAA) the CRLF. 5.2.3 Indirect Effects (via Habitat Effects) 5.2.3.1 Aquatic Plants (Vascular and Non-vascular) Aquatic plants serve several important functions in aquatic ecosystems. Non-vascular aquatic plants are primary producers and provide the autochthonous energy base for aquatic ecosystems. In addition to energy, vascular plants provide structure as attachment sites and refugia for many aquatic invertebrates, fish, and juvenile organisms, such as fish and frogs. In addition, vascular plants also provide primary productivity and oxygen to the aquatic ecosystem. Rooted plants help reduce sediment loading and provide stability to nearshore areas and lower streambanks. In addition, vascular aquatic plants are important as attachment sites for egg masses of CRLFs. Potential indirect effects to the CRLF based on impacts to habitat and/or primary production were assessed using RQs from freshwater aquatic vascular and non-vascular plant data. Indirect effects of diuron to the aquatic-phase CRLF (tadpoles) via reduction in non- vascular aquatic plants in its diet are based on peak EECs from the standard pond and the lowest acute toxicity value for aquatic non-vascular plants. The Agency's LOC (1.0) is exceeded for all current uses of diuron in California. The RQs range from 2331.67 (paved areas) to 1.82 (sorghum). Indirect effects to the CRLF via direct toxicity to aquatic plants are estimated using the most sensitive non-vascular and vascular plant toxicity endpoints. The RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses of diuron in California. These range from 373.33 (paved areas) to 1.01 (alfalfa aerial). Based on LOC exceedances in vascular and non-vascular plants, diuron is Likely to Adversely Affect (LAA) the CRLF indirectly via habitat degradation through reduction in vascular and non-vascular aquatic plants. 5.2.3.2 Terrestrial Plants Terrestrial plants serve several important habitat-related functions for the CRLF. In addition to providing habitat and cover for invertebrate and vertebrate prey items of the CRLF, terrestrial vegetation also provides shelter for the CRLF and cover from predators while foraging. Terrestrial plants also provide energy to the terrestrial ecosystem through primary production. Upland vegetation including grassland and woodlands provides 108 ------- cover during dispersal. Riparian vegetation helps to maintain the integrity of aquatic systems by providing bank and thermal stability, serving as a buffer to filter out sediment, nutrients, and contaminants before they reach the watershed, and serving as an energy source. Twenty-two incidents have been reported that were either registered use or of undetermined legality that resulted in plant damage. The RQs for non-target terrestrial monocot and dicot plants inhabiting semi-aquatic and upland dry areas exceed the Agency's LOC (1.0) for all uses except for monocot plants exposed to sorghum applications. These exceedances range from 300.00 (semi-aquatic plants exposed to non- agricultural aerial applications) to 0.19 (dicot plants inhabiting dry areas exposed to sorghum applications). Several diuron uses result in LOC exceedances from spray drift. These exceedances range from 12.00 (dicot plants exposed to field corn applications) to 1.13 (monocot plants exposed to cotton applications). Based on LOC exceedances in vascular and non-vascular plants, and twenty-two reported incidents that resulted in plant damage, diuron is Likely to Adversely Affect (LAA) the CRLF indirectly via habitat degradation through reduction terrestrial plants. 5.2.4 Modification to Designated Critical Habitat Risk conclusions for the designated critical habitat are the same as those for indirect effects. Agency concludes that there is a potential indirect impact to CRLF by terrestrial habitat degradation from diuron exposure. 5.2.4.1 Aquatic-Phase PCEs Three of the four assessment endpoints for the aquatic-phase primary constituent elements (PCEs) of designated critical habitat for the CRLF are related to potential effects to aquatic and/or terrestrial plants: • Alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond: aquatic habitat (including riparian vegetation) provides for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. • Alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food source. • Reduction and/or modification of aquatic-based food sources for pre-metamorphs (e.g., algae). Based on the risk estimation for potential effects to aquatic and/or terrestrial plants provided in Sections 5.1.1.2, 5.1.1.3, and 5.1.2.3, diuron will result in habitat modification based on effects to aquatic-phase PCEs of designated critical habitat. • Aquatic non-vascular plants used as food source and habitat for CRLF may be potentially affected from all diuron uses. 109 ------- • Reduction of aquatic based food sources may occur from most use sites. • Due to aquatic vascular and terrestrial plant communities being reduced from most use sites, there is potential for alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond. • Due to aquatic vascular and terrestrial plant communities being reduced from most use sites, there is potential for alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food. The remaining aquatic-phase PCE is "alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source." To assess the impact of diuron on this PCE, acute and chronic freshwater fish and invertebrate toxicity endpoints, as well endpoints for aquatic non-vascular plants are used as measures of effects. RQs for these endpoints were calculated in Sections 5.1.1.1 and 5.1.1.2. Based on acute and chronic LOC exceedances for freshwater fish and aquatic invertebrates, diuron will result in habitat modification based on effects to aquatic- phase PCEs of designated critical habitat related to effects of alteration of other chemical characteristics necessary for normal growth and viability of CRLFs and their food source. 5.2.4.2 Terrestrial-Phase PCEs Two of the four assessment endpoints for the terrestrial-phase PCEs of designated critical habitat for the CRLF are related to potential effects to terrestrial plants: • Elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs: Upland areas within 200 ft of the edge of the riparian vegetation or drip line surrounding aquatic and riparian habitat that are comprised of grasslands, woodlands, and/or wetland/riparian plant species that provides the CRLF shelter, forage, and predator avoidance. • Elimination and/or disturbance of dispersal habitat: Upland or riparian dispersal habitat within designated units and between occupied locations within 0.7 mi of each other that allow for movement between sites including both natural and altered sites which do not contain barriers to dispersal. There is a potential for habitat modification via impacts to terrestrial plants (Section 5.2.3.2). The risk estimation for terrestrial-phase PCEs of designated habitat related to potential effects on terrestrial plants is provided in Section 5.1.2.3. These results will inform the effects determination for modification of designated critical habitat for the CRLF. The third terrestrial-phase PCE is "reduction and/or modification of food sources for terrestrial phase juveniles and adults." To assess the impact of diuron on this PCE, acute 110 ------- and chronic toxicity endpoints for birds, mammals, and terrestrial invertebrates are used as measures of effects. RQs for these endpoints were calculated in Section 5.1.2.2. Based on acute and chronic LOC exceedances for CRLF prey items of small mammals, terrestrial invertebrates and other frogs, diuron will result in habitat modification based on effects to the terrestrial PCE relative to reduction in food sources. The fourth terrestrial-phase PCE is based on alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. Direct acute and chronic RQs for terrestrial-phase CRLFs are presented in Section 5.2.1.2. Due to acute and chronic LOC exceedances at all use sites to terrestrial-phase CRLFs, diuron will result in habitat modification based on effects to the terrestrial PCE related to alteration of chemical characteristics necessary for normal growth and viability. 5.2.5.1 Downstream Dilution The downstream extent of exposure in streams and rivers where the EEC could potentially be above levels that would exceed the most sensitive LOC. To complete this assessment, the greatest ratio of aquatic RQ to LOC was estimated. Using an assumption of uniform runoff across the landscape, it is assumed that streams flowing through treated areas (i.e. the initial area of concern) are represented by the modeled EECs; as those waters move downstream, it is assumed that the influx of non-impacted water will dilute the concentrations of diuron present. Using a EC50 value of 2.40 ug/L for non-vascular aquatic plants (the most sensitive species) and a maximum peak EEC for applications to Agricultural Rights-of-Way, Fencerows, etc. of 688 ug/L yields an RQ/LOC ratio of 142 (142/1). Using the downstream dilution approach (described in more detail in Appendix F) yields a distance of 285 kilometers which represents the maximum continuous distance of downstream dilution from the edge of the initial area of concern. Similar to the spray drift buffer described above, the LAA/NLAA determination is based on the area defined by the point where concentrations exceed the EC50 value. 6.0 Uncertainties 6.1. Exposure Assessment Uncertainties 6.1.1 Maximum Use Scenario The screening-level risk assessment focuses on characterizing potential ecological risks resulting from a maximum use scenario, which is determined from labeled statements of maximum application rate and number of applications with the shortest time interval between applications. The frequency at which actual uses approach this maximum use scenario may be dependant on pest resistance, timing of applications, cultural practices, and market forces. Ill ------- 6.1.2 Aquatic Exposure Modeling of Diuron The standard ecological water body scenario (EXAMS pond) used to calculate potential aquatic exposure to pesticides is intended to represent conservative estimates, and to avoid underestimations of the actual exposure. The standard scenario consists of application to a 10-hectare field bordering a 1-hectare, 2-meter deep (20,000 m3) pond with no outlet. Exposure estimates generated using the EXAMS pond are intended to represent a wide variety of vulnerable water bodies that occur at the top of watersheds including prairie pot holes, playa lakes, wetlands, vernal pools, man-made and natural ponds, and intermittent and lower order streams. As a group, there are factors that make these water bodies more or less vulnerable than the EXAMS pond. Static water bodies that have larger ratios of pesticide-treated drainage area to water body volume would be expected to have higher peak EECs than the EXAMS pond. These water bodies will be either smaller in size or have larger drainage areas. Smaller water bodies have limited storage capacity and thus may overflow and carry pesticide in the discharge, whereas the EXAMS pond has no discharge. As watershed size increases beyond 10-hectares, it becomes increasingly unlikely that the entire watershed is planted with a single crop that is all treated simultaneously with the pesticide. Headwater streams can also have peak concentrations higher than the EXAMS pond, but they likely persist for only short periods of time and are then carried and dissipated downstream. The Agency acknowledges that there are some unique aquatic habitats that are not accurately captured by this modeling scenario and modeling results may, therefore, under- or over-estimate exposure, depending on a number of variables. For example, aquatic-phase CRLFs may inhabit water bodies of different size and depth and/or are located adjacent to larger or smaller drainage areas than the EXAMS pond. The Agency does not currently have sufficient information regarding the hydrology of these aquatic habitats to develop a specific alternate scenario for the CRLF. CRLFs prefer habitat with perennial (present year-round) or near-perennial water and do not frequently inhabit vernal (temporary) pools because conditions in these habitats are generally not suitable (Hayes and Jennings 1988). Therefore, the EXAMS pond is assumed to be representative of exposure to aquatic-phase CRLFs. In addition, the Services agree that the existing EXAMS pond represents the best currently available approach for estimating aquatic exposure to pesticides (USFWS/NMFS 2004). In general, the linked PRZM/EXAMS model produces estimated aquatic concentrations that are expected to be exceeded once within a ten-year period. The Pesticide Root Zone Model is a process or "simulation" model that calculates what happens to a pesticide in an agricultural field on a day-to-day basis. It considers factors such as rainfall and plant transpiration of water, as well as how and when the pesticide is applied. It has two major components: hydrology and chemical transport. Water movement is simulated by the use of generalized soil parameters, including field capacity, wilting point, and saturation water content. The chemical transport component can simulate pesticide application on the soil or on the plant foliage. Dissolved, adsorbed, and vapor-phase concentrations in the soil are estimated by simultaneously considering the processes of pesticide uptake by 112 ------- plants, surface runoff, erosion, decay, volatilization, foliar wash-off, advection, dispersion, and retardation. Uncertainties associated with each of these individual components add to the overall uncertainty of the modeled concentrations. Additionally, model inputs from the environmental fate degradation studies are chosen to represent the upper confidence bound on the mean values that are not expected to be exceeded in the environment approximately 90 percent of the time. Mobility input values are chosen to be representative of conditions in the environment. The natural variation in soils adds to the uncertainty of modeled values. Factors such as application date, crop emergence date, and canopy cover can also affect estimated concentrations, adding to the uncertainty of modeled values. Factors within the ambient environment such as soil temperatures, sunlight intensity, antecedent soil moisture, and surface water temperatures can cause actual aquatic concentrations to differ for the modeled values. Unlike spray drift, tools are currently not available to evaluate the effectiveness of a vegetative setback on runoff and loadings. The effectiveness of vegetative setbacks is highly dependent on the condition of the vegetative strip. For example, a well- established, healthy vegetative setback can be a very effective means of reducing runoff and erosion from agricultural fields. Alternatively, a setback of poor vegetative quality or a setback that is channelized can be ineffective at reducing loadings. Until such time as a quantitative method to estimate the effect of vegetative setbacks on various conditions on pesticide loadings becomes available, the aquatic exposure predictions are likely to overestimate exposure where healthy vegetative setbacks exist and underestimate exposure where poorly developed, channelized, or bare setbacks exist. In order to account for uncertainties associated with modeling, available monitoring data were compared to PRZM/EXAMS estimates of peak EECs for the different uses. As discussed above, several data values were available from NAWQA for diuron concentrations measured in surface waters receiving runoff from agricultural areas. The specific use patterns (e.g. application rates and timing, crops) associated with the agricultural areas are unknown, however, they are assumed to be representative of potential diuron use areas. The monitoring data available are below the PZM-EXAMS predictions by an order of magnitude. The monitoring data indicated that the time weighted mean concentration ranged from below the 0.001 ppb to 0.86 ppb with a mean concentration of 0.21 ppb. PRZM-EXAMS prediction (not including the scenarios for impervious surfaces) estimated diuron to be in the surface waters at peak concentrations from 27.5 ppb to 2.6 ppb and 60 day EEC from 18.3 ppb to 1.8. 6.1.3 Action Area Uncertainties The action area is considered to be the whole State of California since diuron is considered to be an animal carcinogen and can not be spatially defined. Diuron use has been reported in all 58 counties. 113 ------- 6.1.4 Usage Uncertainties County-level usage data were obtained from California's Department of Pesticide Regulation Pesticide Use Reporting (CDPR PUR) database. Four years of data (2002 - 2005) were included in this analysis because statistical methodology for identifying outliers, in terms of area treated and pounds applied, was provided by CDPR for these years only. No methodology for removing outliers was provided by CDPR for 2001 and earlier pesticide data; therefore, this information was not included in the analysis because it may misrepresent actual usage patterns. CDPR PUR documentation indicates that errors in the data may include the following: a misplaced decimal; incorrect measures, area treated, or units; and reports of diluted pesticide concentrations. In addition, it is possible that the data may contain reports for pesticide uses that have been cancelled. The CPDR PUR data does not include home owner applied pesticides; therefore, residential uses are not likely to be reported. As with all pesticide usage data, there may be instances of misuse and misreporting. The Agency made use of the most current, verifiable information; in cases where there were discrepancies, the most conservative information was used. 6.1.5 Terrestrial Exposure Modeling of Diuron The Agency relies on the work of Fletcher et al. (1994) for setting the assumed pesticide residues in wildlife dietary items. These residue assumptions are believed to reflect a realistic upper-bound residue estimate, although the degree to which this assumption reflects a specific percentile estimate is difficult to quantify. It is important to note that the field measurement efforts used to develop the Fletcher estimates of exposure involve highly varied sampling techniques. It is entirely possible that much of these data reflect residues averaged over entire above ground plants in the case of grass and forage sampling. It was assumed that ingestion of food items in the field occurs at rates commensurate with those in the laboratory. Although the screening assessment process adjusts dry- weight estimates of food intake to reflect the increased mass in fresh-weight wildlife food intake estimates, it does not allow for gross energy differences. Direct comparison of a laboratory dietary concentration- based effects threshold to a fresh-weight pesticide residue estimate would result in an underestimation of field exposure by food consumption by a factor of 1.25 - 2.5 for most food items. Differences in assimilative efficiency between laboratory and wild diets suggest that current screening assessment methods do not account for a potentially important aspect of food requirements. Depending upon species and dietary matrix, bird assimilation of wild diet energy ranges from 23 - 80%, and mammal's assimilation ranges from 41 - 85% (U.S. Environmental Protection Agency, 1993). If it is assumed that laboratory chow is formulated to maximize assimilative efficiency (e.g., a value of 85%), a potential for underestimation of exposure may exist by assuming that consumption of food in the wild is comparable with consumption during laboratory testing. In the screening process, 114 ------- exposure may be underestimated because metabolic rates are not related to food consumption. For the terrestrial exposure analysis of this risk assessment, a generic bird or mammal was assumed to occupy either the treated field or adjacent areas receiving a treatment rate on the field. Actual habitat requirements of any particular terrestrial species were not considered, and it was assumed that species occupy, exclusively and permanently, the modeled treatment area. Spray drift model predictions suggest that this assumption leads to an overestimation of exposure to species that do not occupy the treated field exclusively and permanently. 6.1.6 Spray Drift Modeling It is unlikely that the same organism would be exposed to the maximum amount of spray drift from every application made. In order for an organism to receive the maximum concentration of diuron from multiple applications, each application of diuron would have to occur under identical atmospheric conditions (e.g., same wind speed and same wind direction) and (if it is an animal) the animal being exposed would have to be located in the same location (which receives the maximum amount of spray drift) after each application. Additionally, other factors, including variations in topography, cover, and meteorological conditions over the transport distance are not accounted for by the AgDRIFT model (i.e., it models spray drift from aerial and ground applications in a flat area with little to no ground cover and a steady, constant wind speed and direction). Therefore, in most cases, the drift estimates from AgDRIFT may overestimate exposure, especially as the distance increases from the site of application, since the model does not account for potential obstructions (e.g., large hills, berms, buildings, trees, etc.). Furthermore, conservative assumptions are made regarding the droplet size distributions being modeled (was set to the default' ASAE Very Fine to Fine'), the application method (i.e., aerial), release heights and wind speeds (also set to the default values of 10 ft and 10 mph respectively for aerial application; the height for ground application was set to the default 'high boom'). Alterations in any of these inputs would decrease the area of potential effect. 6.2 Effects Assessment Uncertainties 6.2.1 Age Class and Sensitivity of Effects Thresholds It is generally recognized that test organism age may have a significant impact on the observed sensitivity to a toxicant. The acute toxicity data for fish are collected on juvenile fish between 0.1 and 5 grams. Aquatic invertebrate acute testing is performed on recommended immature age classes (e.g., first instar for daphnids, second instar for amphipods, stoneflies, mayflies, and third instar for midges). Testing of juveniles may overestimate toxicity at older age classes for pesticide active ingredients that act directly without metabolic transformation because younger age classes may not have the enzymatic systems associated with detoxifying xenobiotics. In 115 ------- so far as the available toxicity data may provide ranges of sensitivity information with respect to age class, this assessment uses the most sensitive life-stage information as measures of effect for surrogate aquatic animals, and is therefore, considered as protective of the CRLF. 6.2.2 Use of Surrogate Species Effects Data CRLF Guideline toxicity tests and open literature data on diuron are not available for frogs or any other aquatic-phase amphibian; therefore, freshwater fish are used as surrogate species for aquatic-phase amphibians. Therefore, endpoints based on freshwater fish ecotoxicity data are assumed to be protective of potential direct effects to aquatic-phase amphibians including the CRLF, and extrapolation of the risk conclusions from the most sensitive tested species to the aquatic-phase CRLF is likely to overestimate the potential risks to those species. Efforts are made to select the organisms most likely to be affected by the type of compound and usage pattern; however, there is an inherent uncertainty in extrapolating across phyla. In addition, the Agency's LOCs are intentionally set very low, and conservative estimates are made in the screening level risk assessment to account for these uncertainties. Terrestrial Plants Plant toxicity data from both registrant-submitted studies and studies in the scientific literature were reviewed for this assessment. Registrant-submitted studies are conducted under conditions and with species defined in EPA toxicity test guidelines. Sub-lethal endpoints such as plant growth, dry weight, and biomass are evaluated for both monocots and dicots, and effects are evaluated at both seedling emergence and vegetative life stages. Guideline studies generally evaluate toxicity to ten crop species. A drawback to these tests is that they are conducted on herbaceous crop species only, and extrapolation of effects to other species, such as the woody shrubs and trees and wild herbaceous species, contributes uncertainty to risk conclusions. Commercial crop species have been selectively bred, and may be more or less resistant to particular stressors than wild herbs and forbs. The direction of this uncertainty for specific plants and stressors, including diuron, is largely unknown. Homogenous test plant seed lots also lack the genetic variation that occurs in natural populations, so the range of effects seen from tests is likely to be smaller than would be expected from wild populations. 116 ------- 6.2.3 Sublethal Effects When assessing acute risk, the screening risk assessment relies on the acute mortality endpoint as well as a suite of sublethal responses to the pesticide, as determined by the testing of species response to chronic exposure conditions and subsequent chronic risk assessment. Consideration of additional sublethal data in the effects determination t is exercised on a case-by-case basis and only after careful consideration of the nature of the sublethal effect measured and the extent and quality of available data to support establishing a plausible relationship between the measure of effect (sublethal endpoint) and the assessment endpoints. However, the full suite of sublethal effects from valid open literature studies is considered for the purposes of defining the action area. 6.2.4 Location of Wildlife Species For the terrestrial exposure analysis of this risk assessment, a generic bird or mammal was assumed to occupy either the treated field or adjacent areas receiving a treatment rate on the field. Actual habitat requirements of any particular terrestrial species were not considered, and it was assumed that species occupy, exclusively and permanently, the modeled treatment area. Spray drift model predictions suggest that this assumption leads to an overestimation of exposure to species that do not occupy the treated field exclusively and permanently. 7.0 Risk Conclusions In fulfilling its obligations under Section 7(a) (2) of the Endangered Species Act, the information presented in this endangered species risk assessment represents the best data currently available to assess the potential risks of diuron to the CRLF and its designated critical habitat. The Agency makes a Likely to Adversely Affect determination for the CRLF from the use of diuron. The Agency has determined that there is the potential for modification of CRLF designated critical habitat from the use of the chemical. The direct effects and habitat modification effects determinations are summarized in Table 7.1 and Table 7.2 respectively. Given the LAA determination for the CRLF and potential modification of designated critical habitat, a description of the baseline status and cumulative effects for the CRLF is provided in Attachment II. The LAA effects determination applies to those areas where it is expected that the pesticide's use will directly or indirectly affect the CRLF or its designated critical habitat. To determine this area, the footprint of iron's use pattern is identified, using land cover data that correspond to iron's use pattern. The spatial extent of the LAA effects determination also includes areas beyond the initial area of concern that may be impacted by runoff and/or spray drift. The identified direct and indirect effects and modification to critical habitat are anticipated to occur only for those currently occupied core habitat areas, CNDDB occurrence sections, and designated critical habitat for the CRLF that overlap with the initial area of concern. The AgDRIFT model was used to evaluate 117 ------- potential distances beyond which exposures would be expected to be below LOC. However, due to the limitations imposed by the Tier 1 ground analysis (allows users to evaluate off-site deposition and exposure out to 1,000 ft downwind from the location of the application), the exact buffer needed for exposures to be below the LOC is uncertain. The output from AgDRIFT indicated that the buffer zone required would be greater than 1,000 feet. Since the model is restricted to accurately discerning a buffer within 1,000 feet of the application, the exact distance needed for a buffer to protect non-listed and listed plants is unknown. However, as seen in Table 3.7, the calculated risk quotient (RQ) is significantly larger than the level of concern (LOC) for non-listed and listed plant species (LOC = 1). This comparison provides awareness to approximately how much greater than 1,000 feet the buffer needs to be. It is assumed that non-flowing water bodies (or potential CRLF habitat) are included within this area. In addition to the spray drift buffer, the results of the downstream dilution extent analysis result in a distance of 285 kilometers which represents the maximum continuous distance of downstream dilution from the edge of the initial area of concern. If any of these streams reaches flow into CRLF habitat, there is potential to affect either the CRLF or modify its habitat. These lotic aquatic habitats within the CRLF core areas and critical habitats potentially contain concentrations of diuron sufficient to result in LAA determination or modification of critical habitat. Appendix F provides maps of the initial area of concern, along with CRLF habitat areas, including currently occupied core areas, CNDDB occurrence sections, and designated critical habitat. It is expected that any additional areas of CRLF habitat that are located 285 kilometers of stream distance (to account for downstream dilution) outside the initial area of concern may also be impacted and are part of the full spatial extent of the LAA/modification of critical habitat effects determination. 118 ------- Table 7.1 El Tects Determination Summary for Effects of Diuron on the CRLF Assessment Endpoint Effects Determination 1 Basis for Determination Survival, growth, and/or reproduction ofCRLF individuals LAA Potential for Direct Effects Aquatic-phase (Eggs, Larvae, and Adults): The aquatic phase amphibian acute LOCs for Listed species (0.05) are exceeded for most uses of diuron in California. Acute RQs that exceed the Agency's LOC range from 13.99 (paved areas) to 0.053 (papaya). Chronic RQs that exceed the Agency's LOC for chronic exposure (1.0) range from 188.58 (paved areas) to 1.0003 (banana, plantain). Terrestrial-phase (Juveniles and Adults): Refined acute dietary-based RQs for CRLFs consuming large insects and small herbivore mammals exceed the acute listed species LOC (0.1) for all uses of diuron except Sorghum and Field Corn. The acute dietary-based RQs for CRLFs consuming large insects exceed the acute listed species LOC for Agricultural Rights-of-Way, Fencerows etc. and Ornamental Herbaceous Plants. No acute dietary based LOCs were exceeded for CLRF consuming small insectivore mammals and small terrestrial phase amphibians for any diuron use. Refined chronic dietary-based RQs for CRLFs consuming insects and mammals exceed the chronic listed species LOC for all uses of diuron across all food item groups. Potential for Indirect Effects Aquatic prey items, aquatic habitat, cover and/or primary productivity. LOCs for non-vascular plants are exceeded for all uses. The RQs range from 2331.67 (paved areas) to 1.82 (sorghum). Aquatic invertebrates acute a LOC are exceeded. The acute RQs that exceed the LOC range from 34.98 (paved areas) to 0.06 (post-harvest wheat). Chronic LOCs are exceeded for paved area applications (RQ=1.50). RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 373.33 (paved areas) to 1.01 (alfalfa aerial). Terrestrial prey items, riparian habitat. For small mammal, chronic dose based RQs exceed the Agency's LOC (1.0) for most uses. The dose based chronic RQs that exceed the LOC range from 5.00 (paved areas) to 1.31 (walnut). The acute RQs exceed the Agency' s LOC for several uses. The RQs that exceed the acute dose based LOC range from 0.25 (paved areas) to 0.12 (Agricultural Rights-of-Way, Fencerows, Hedgerows, Airports, etc.). LOCs are exceeded for terrestrial riparian plants and for aquatic plants from exposure to diuron from runoff or spray drift. Alteration of riparian and vascular plants may result in alteration of temperature, turbidity, and oxygen content. RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses. These range from 373.33 (paved areas) to 1.01 (alfalfa aerial). 1 No effect (NE); May affect, but not likely to adversely affect (NLAA); May affect, likely to adversely affect (LAA) 119 ------- Table 7.2 Effects Determination Summary for the Critical Habitat Impact Analysis Assessment Endpoint Effects Determination 1 Basis for Determination Modification of aquatic-phase PCE HM Due to aquatic vascular and terrestrial plant communities being reduced from all use sites, there is potential for alteration of channel/pond morphology or geometry and/or increase in sediment deposition within the stream channel or pond. These plant communities provide for shelter, foraging, predator avoidance, and aquatic dispersal for juvenile and adult CRLFs. In addition, there is potential for alteration in water chemistry/quality including temperature, turbidity, and oxygen content necessary for normal growth and viability of juvenile and adult CRLFs and their food. LOCs are exceeded for terrestrial riparian plants and for aquatic vascular plants from exposure to diuron from runoff or spray drift. LOCs for non-vascular plants are exceeded for all uses. Modification of terrestrial-phase PCE HM The use of diuron at all use sites may create the following modification of PCE: elimination and/or disturbance of upland habitat; ability of habitat to support food source of CRLFs, Elimination and/or disturbance of dispersal habitat, reduction and/or modification of food sources for terrestrial phase juveniles and adults, and alteration of chemical characteristics necessary for normal growth and viability of juvenile and adult CRLFs and their food source. The RQs for vascular aquatic plants exceed the Agency's LOC (1.0) for most uses of diuron in California. These range from 373.33 (paved areas) to 1.01 (alfalfa aerial). Use of diuron on most use sites will exceed acute dietary- and dose-based LOC and chronic LOC for prey food items of small mammals, frogs, and invertebrates. Food source for CRLF is reduced and CRLF is indirectly affected. 1 Habitat Modification (HM) or No effect (NE) Based on the conclusions of this assessment, a formal consultation with the U. S. Fish and Wildlife Service under Section 7 of the Endangered Species Act should be initiated. When evaluating the significance of this risk assessment's direct/indirect and adverse habitat modification effects determinations, it is important to note that pesticide exposures and predicted risks to the species and its resources (i.e., food and habitat) are not expected to be uniform across the action area. In fact, given the assumptions of drift and downstream transport (i.e., attenuation with distance), pesticide exposure and associated risks to the species and its resources are expected to decrease with increasing distance away from the treated field or site of application. Evaluation of the implication of this non-uniform distribution of risk to the species would require information and assessment techniques that are not currently available. Examples of such information and methodology required for this type of analysis would include the following: • Enhanced information on the density and distribution of CRLF life stages within specific recovery units and/or designated critical habitat within the action area. This information would allow for quantitative extrapolation of the present risk assessment's predictions of individual effects to the 120 ------- proportion of the population extant within geographical areas where those effects are predicted. Furthermore, such population information would allow for a more comprehensive evaluation of the significance of potential resource impairment to individuals of the species. • Quantitative information on prey base requirements for individual aquatic- and terrestrial-phase frogs. While existing information provides a preliminary picture of the types of food sources utilized by the frog, it does not establish minimal requirements to sustain healthy individuals at varying life stages. Such information could be used to establish biologically relevant thresholds of effects on the prey base, and ultimately establish geographical limits to those effects. This information could be used together with the density data discussed above to characterize the likelihood of adverse effects to individuals. • Information on population responses of prey base organisms to the pesticide. Currently, methodologies are limited to predicting exposures and likely levels of direct mortality, growth or reproductive impairment immediately following exposure to the pesticide. The degree to which repeated exposure events and the inherent demographic characteristics of the prey population play into the extent to which prey resources may recover is not predictable. An enhanced understanding of long-term prey responses to pesticide exposure would allow for a more refined determination of the magnitude and duration of resource impairment, and together with the information described above, a more complete prediction of effects to individual frogs and potential modification to critical habitat. 121 ------- 8.0 References Altig, R. and R.W. McDiarmid. 1999. Body Plan: Development and Morphology. In R.W. McDiarmid and R. Altig (Eds.), Tadpoles: The Biology of Anuran Larvae.University of Chicago Press, Chicago, pp. 24-51. Alvarez, J. 2000. 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