Aquatic Pesticide Application Plan for the San Francisco Estuary Invasive Spartina Project

This plan addresses herbicide application activities undertaken by ISP Partner agencies in the effort to eradicate non-native, invasive Spartina from the San Francisco Estuary.

Annual update prepared by Drew Kerr 2612-A 8th Street Berkeley, CA 94710 [email protected]

for the State Coastal Conservancy 1330 Broadway, 13th floor Oakland, Ca 94612-2530

August 2010

Current funding for the San Francisco Estuary Invasive Spartina Project comes from the California State Coastal Conservancy and grants from the U.S. Environmental Protec- tion Agency, the National Oceanic and Atmospheric Administration and the U.S. Miner- als Management Service. Previous major funders include the CALFED Bay-Delta Pro- gram, the California Wildlife Conservation Board, the U.S. Fish and Wildlife Service and the National Fish and Wildlife Foundation. Table of Contents

1. BACKGROUND ...... 1 2. STATEMENT OF PURPOSE AND NEED...... 1 3. DESCRIPTION OF THE WATER BODY SYSTEM...... 2 Ecology...... 2 Natural Processes Affecting Water Quality...... 3 Water Quality...... 4 Sediment Quality ...... 7 4. DESCRIPTION OF TARGET SPECIES...... 9 Native Pacific Cordgrass (Spartina foliosa)...... 9 Atlantic Smooth Cordgrass (Spartina alterniflora) and its Hybrids...... 10 English Cordgrass (Spartina anglica) ...... 13 Chilean Cordgrass (Spartina densiflora)...... 13 Salt-Meadow Cordgrass (Spartina patens) ...... 14 5. CONTROL TOLERANCES ...... 15 6. DESCRIPTION OF HERBICIDE...... 15 Environmental Fate of Herbicides ...... 20 Potential Biological and Ecological Effects ...... 25 Non-Target Aquatic Plants and Algae...... 27 Aquatic and Benthic Invertebrates ...... 28 Fish...... 30 Birds ...... 31 Mammals...... 31 7. DESCRIPTION OF ALTERNATE, NON-CHEMICAL CONTROL METHODS.. 32 ISP-selected Non-chemical Control Methods...... 33 Hand-pulling and manual excavation...... 33 Mechanical excavation and dredging ...... 33 Covering/blanketing ...... 34 Other Non-chemical Methods Evaluated...... 35 Mowing, burning, pruning, and flaming ...... 35 Crushing and mechanical smothering ...... 36 Flooding and draining ...... 36 8. HERBICIDE APPLICATION AREAS FOR 2008-2010...... 37 Site 01 – Alameda Flood Control Channel, Alameda County...... 37 Site 02 – Bair & Greco Island Complex, San Mateo County ...... 39 Site 03 – Blackie’s Pasture, Marin County...... 39 Site 04 – Corte Madera Creek Complex, Marin County ...... 40 Site 05 – Coyote Creek & Mowry Slough Area, Santa Clara County...... 40 Site 06 – Emeryville Crescent, Alameda County ...... 41 Site 07 – Oro Loma Marsh, Alameda County ...... 41 Site 08 – Palo Alto Baylands, Santa Clara County...... 42 Site 09 – Tiscornia Marsh (formerly Pickleweed Park), Marin County ...... 42 Site 10 – Point Pinole Regional Shoreline, Contra Costa County ...... 43 Site 11 – Southampton Marsh, Solano County...... 43 Site 12 – Southeast San Francisco, San Francisco County...... 44 Site 13 – Whale’s Tail / Old Alameda Creek Complex, Alameda County...... 44 Site 15 – South San Francisco Bay Marshes, Santa Clara County ...... 45 Site 16 – Cooley Landing, San Mateo County ...... 46

San Francisco Estuary Invasive Spartina Project i. 2010 Aquatic Pesticide Application Plan Site 17 – Alameda Island / San Leandro Bay Complex, Alameda County ...... 46 Site 18 – Colma Creek / San Bruno Marsh Complex, San Mateo County ...... 47 Site 19 – West San Francisco Bay, San Mateo County ...... 48 Site 20 – San Leandro / Hayward Shoreline Complex, Alameda County ...... 49 Site 21 – Ideal Marsh, Alameda County...... 49 Site 22 – Two Points Complex, Alameda County ...... 50 Site 23 – Marin Outliers, Marin County...... 50 Site 24 – Petaluma River, Sonoma County...... 51 Site 26 – North San Pablo Bay, Solano County ...... 52 9. WATER QUALITY MONITORING PLAN ...... 52 Objective...... 52 Monitoring Site Selection ...... 52 Sampling Design...... 53 Field Sampling Procedures...... 53 Equipment Calibration ...... 54 Field Data Sheets ...... 55 Sample Shipment ...... 55 Field Variances ...... 55 Sample Analysis...... 55 Assessment of Field Contamination...... 56 Lab QC & Data Quality Indicators ...... 56 Monitoring Site Descriptions...... 56 10. APPLICABLE WATER QUALITY BMPS ...... 61 11. REFERENCES...... 63

List of Figures Figure 1: Locations and mean discharges for municipal wastewater treatment plants in South San Francisco Bay...... 4 Figure 2: Conceptual Model of Possible Exposure of Biological Organisms to Herbicide Mixture Used by the Spartina Control Program...... 26 Figure 3: Baywide map of 2010 Spartina Control Program treatment sites ...... 38

List of Tables Table 1: Dissolved concentrations of trace metals in water samples...... 7 Table 2: Total concentrations of trace metals in water samples ...... 7 Table 3: Ranges of trace pollutants in San Francisco Bay sediments...... 8 Table 4a: Imazapyr herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Estuary ...... 21 Table 4b: Glyphosate herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Estuary ...... 21 Table 5: Summary of water quality monitoring sites for the 2010 season...... 54

San Francisco Estuary Invasive Spartina Project ii. 2010 Aquatic Pesticide Application Plan Appendices 1. Chemical properties, degradation rates, environmental fate, and toxicity of imazapyr, glyphosate, and aquatic surfactants evaluated for Spartina control 2. 2010 Spartina Control Program Site Maps 3. Field Data Collection Form 4. Chain of Custody form 5. Laboratory Quality Assurance Plan, Pacific Agricultural Laboratory 6. General Site Safety & Materials Handling Guidelines and Procedures for Spartina Control Projects in the San Francisco Estuary

San Francisco Estuary Invasive Spartina Project iii. 2010 Aquatic Pesticide Application Plan 1. BACKGROUND The San Francisco Bay Estuary (San Francisco Estuary or Estuary) supports the largest and most ecologically important expanses of tidal mudflats and salt marshes in the contiguous western United States. This environment naturally supports a diverse array of native plants and animals. Over the years, many non-native species of plants and animals have been introduced to the Estuary, and some now threaten to cause fundamental changes in the structure, function, and value of the Estuary's tidal lands. Among these threatening invaders are several species of salt marsh cordgrass (genus Spartina). In recent decades, populations of non-native cordgrasses were introduced to the Estuary and began to spread rapidly. Though valuable in their native settings, these introduced cordgrasses are highly aggressive in this new environment, and frequently become the dominant plant species in areas they invade. One of the non-native cordgrass species, Atlantic smooth cordgrass (Spartina alterniflora), is rapidly spreading throughout the Estuary, particularly in the South San Francisco Bay (South Bay). Atlantic smooth cordgrass and its hybrids (formed when this species crosses with the native Pacific cordgrass, S. foliosa) are now threatening the ecological balance of the Estuary. Based on a century of international studies of comparable cordgrass invasions, these hybrids are likely to eventually cause the extinction of native Pacific cordgrass, choke tidal creeks, dominate newly restored tidal marshes, and displace thousands of acres of existing shorebird habitat. Once established in this estuary, invasive cordgrasses could rapidly spread to other estuaries along the California coast through seed dispersal on the tides. Non-native invasive cordgrasses currently dominate approximately 1,200-1,500 acres of the San Francisco Estuary in seven counties — on State, Federal, municipal, and private lands— and are spreading at an alarming rate. The California State Coastal Conservancy (Conservancy) initiated the ISP in 2000 to stave off the invasion of non-native cordgrass and its potential impacts. The ISP is a regionally coordinated effort of Federal, State, and local agencies, private landowners, and other interested parties, with the ultimate goal of arresting and reversing the spread of non-native cordgrasses in the San Francisco Estuary. When fully implemented, the ISP will provide opportunities to maximize resources, effectively disseminate information, facilitate regional monitoring, and reduce the occurrence of cordgrass re-infestation. The geographic focus of the ISP includes the nearly 40,000 acres of tidal marsh and 29,000 acres of tidal flats that comprise the shoreline areas of the nine Bay Area counties, including Alameda, Contra Costa, Marin, Napa, San Francisco, San Mateo, Santa Clara, So- lano, and Sonoma Counties, and Sacramento County.

2. STATEMENT OF PURPOSE AND NEED The purpose of the Spartina control program is to arrest and reverse the spread of invasive non-native cordgrass species in the San Francisco Estuary to preserve and restore the ecological integrity of the Estuary's intertidal habitats and estuarine ecosystem. The Spartina control program is needed to prevent further degradation and loss of the natural ecological structure and function of the San Francisco Estuary. In the absence of any coordinated and wide-ranging control program, within decades one-quarter to one-

San Francisco Estuary Invasive Spartina Project 1. 2010 Aquatic Pesticide Application Plan half (up to 10,000 acres) of the existing intertidal flats are likely to be replaced with dense, invasive cordgrass marsh, and much of the native diverse salt-marsh vegetation replaced with nearly homogeneous stands of non-native cordgrass. This ecological conversion is likely to alter the structure and function of the Estuary, affecting fisheries, migratory shorebirds and waterfowl, marine mammals, endangered fish, wildlife, and plants, tidal sediment transport, and the rate, pattern, and magnitude of tidal flows. In addition, invasive cordgrasses may impede or preclude plans to restore up to 20,000 acres of diked baylands to native tidal marsh. Arresting and reversing the invasion of non- native cordgrasses may not be feasible once these species have spread and become established, due to the expansive scale of the invasion and the effects of hybridization. To avoid these consequences, the ISP is implementing a regionally coordinated, long-term management program.

3. DESCRIPTION OF THE WATER BODY SYSTEM

Ecology Like most Pacific estuaries, the majority of the intertidal zone of the San Francisco Estu- ary naturally consists of unvegetated tidal flats, or mudflats. Native California tidal marsh vegetation is limited to the upper intertidal zones, above mean sea level in San Francisco and San Pablo Bays. Below mean sea level, waves erode and redeposit the upper layers of bay mud with each tidal cycle. Rich deposits of fine silt and clay from the Sacramento- San Joaquin Delta have accumulated in the Estuary to form highly productive mudflats, with abundant benthic invertebrates. The mudflats provide a critical source of nutrition and energy for migratory shorebirds and waterfowl, with more than one million shorebirds using the Estuary's mudflats and salt ponds during migration, and over half of the west-coast migratory diving ducks making this estuary their winter home. At elevations above the intertidal zone (in areas that have not been diked and removed from tidal action), are the Estuary's tidal salt and brackish marshes. Pacific salt marsh vegetation is more diverse in plant species than its Atlantic counterparts. Until recent decades, the native Pacific cordgrass exclusively occupied the lower reaches of the Estu- ary's tidal salt marshes. At slightly higher elevations, a relatively flat tidal marsh plain (reaching near the average level of the higher daily tides), is dominated by low-growing, mostly perennial plants such as pickleweed, saltgrass, and other salt-tolerant herbs. The tidal marsh plain is punctuated by salty shallow ponds (pans), and dissected by irregular tidal creeks. Above the tidal marsh plain, at the uppermost edges of the marsh, are an even greater number of plant species. Many endemic (unique to the area) plant and animal species, including many rare or endangered species, survive only in the Estuary's remaining tidal marshes. They remain at risk of extinction because of the severe decline over the past century in the abundance, distribution, and quality of tidal marshes. Most of the Estuary's rare species have narrow or specific habitat requirements, and the health of their populations usually is sensitive to structural changes in their habitats - particularly the condition of the marsh vegetation. Strong dominance of the vegetation by one or more plant species necessarily results in lower overall species diversity, and can push rarer species to local extinction.

San Francisco Estuary Invasive Spartina Project 2. 2010 Aquatic Pesticide Application Plan Natural Processes Affecting Water Quality Water quality within the San Francisco Estuary is connected to and affected by complex regional and local natural processes. Hydrologic relationships between the Pacific Ocean, the Estuary, and the many freshwater tributaries (including the Sacramento-San Joaquin River system) govern salinity levels in different portions of the Estuary and along the Bay margins. Variable natural factors such as tidal cycles, local winds, basin bathymetry, and salinity gradients interact with river flows and affect the circulation of Estuary waters through channels, Estuary margins, and bays, distributing nutrients, salt concentrations, and pollutants. Major processes affecting water quality are described below. Tidal Cycles. The Estuary has two low tides and two high tides every 24.8 hours. During each tidal cycle, an average of about 1.3 million acre-feet of water, or 24 percent of the Bay and Delta’s volume, moves in and out of the Golden Gate. On the flood (incoming) tide, ocean water moves through the Golden Gate and into the Estuary’s southern and northern reaches, raising the water level at the end of the South Bay by more than eight feet, and raising the height of the Sacramento River at the upstream edge of the Estuary by about three feet. It takes about two hours for the flood tide to reach the end of the South Bay and eight hours to reach Sacramento. Subregional Conditions. Suisun and North Bay subregions receive the majority of freshwater input from the Sacramento and San Joaquin River system. In the open bays, density-driven currents show ebb dominance of the surface water and flood dominance of the bottom water. Waters in these embayments are well oxygenated, with low- to moderate-salinity and high-suspended solids concentrations. Water residence time affects the abundance and distribution of many estuarine organisms, the amount of primary production by phytoplankton, and some of the chemical and physical processes that influence the distribution and fate of pollutants. During low flow periods of the year (late summer), the residence time of freshwater moving from the Delta to the ocean can be relatively long (on the order of months) compared to periods when outflow is very high (winter), when freshwater can move from the Delta to the ocean in days. The Central Bay subregion is influenced by ocean waters that are cold, saline, and lower in total suspended sediment. Water quality parameters fluctuate less than in other sectors of the Bay due to the predominance of ocean water. Net exchanges of ocean and Bay waters depend on freshwater flow in the Bay, tidal amplitude, and longshore coastal currents. The southern part of San Francisco Bay receives less than 10 percent of the natural freshwater flow into the Bay, but the majority (>75 percent) of wastewater discharges. The largest flow is from San Jose, where approximately 120 million gallons per day (MGD) of treated wastewater are released into Artesian Slough, a tributary to Coyote Creek (Figure 1). This fresh water flow creates a local zone of brackish water in the otherwise saline tip if the South Bay. The rest of the South Bay, because it has so little freshwater input, is essentially a tidal lagoon with a relatively constant salinity (approximately the same as ocean water, 32 parts per thousand, ppt). South Bay waters are influenced by Delta outflow only during the winter months, when low-salinity water moves southward into the southern reach displacing the saline, denser water northward. In the summer months, however, South Bay currents are largely influenced by wind stress on

San Francisco Estuary Invasive Spartina Project 3. 2010 Aquatic Pesticide Application Plan the surface; northwest winds transport water in the direction of the wind, and the dis- placed water causes subsurface currents to flow in the opposite direction. Currents and Circulation. Circulation patterns within the Bay are influenced by Delta inflows, gravitational currents, and tide- and wind-induced horizontal circulation. The cumulative effects of the latter three factors on net circulation within embayments tend to dominate over that of freshwater inflows except during short periods after large storm events (Smith 1987). Exchanges between embayments are influenced both by mixing patterns within embayments and by the magnitude of freshwater inflows (Smith 1987). Currents created by tides, freshwater inflows, and winds cause erosion and transport of sediments. Tidal currents are usually the dominant form of observed currents in the Figure 1. Locations and mean discharges for municipal Bay. Tidal currents are stronger in the chan- wastewater treatment plants in South San Francisco nels and weaker in the shallows (Cheng and Bay. Adapted from Schemel et al. 1999, based on Davis Gartner 1984). These processes enhance ex- et al. 1991. change between shallows and channels during the tidal cycle, and contribute significantly to landward mixing of ocean water and seaward mixing of river water. Also, the South Bay begins flooding while San Pablo Bay is still ebbing, making it possible for the South Bay to receive water from the northern reach (Smith 1987). Tides have a significant influence on sediment resuspension during the more energetic spring tide when sediment concentrations naturally increase, and particularly during the ebbs preceding lower low water when the current speeds are highest. Powell et al. (1989), however, observed no correlation between tidal cycle and suspended sediment loads or distribution in the South Bay. Their conclusion was that winds are the most important factor in resuspending sediments in the South Bay, and that sources of sediments are more important than transport of sediment resuspended from other parts of the Bay (Reilly et al. 1992). Wind-induced currents have a significant effect on sediment transport by resuspending sediments in shallow waters (Krone 1979; Cloern et al. 1989). An estimated 100 to 286 million cubic yards of sediments are resuspended annually from shallow areas of the Bay by wind-generated waves (Krone 1974; SFEP 1992b).

Water Quality Water quality in the San Francisco Estuary has improved significantly since the enact- ment of the California Water Quality Control Act (Porter-Cologne) in 1969 and the Clean Water Act in 1972. Nevertheless, the Estuary waters still carry significant loads of pollut-

San Francisco Estuary Invasive Spartina Project 4. 2010 Aquatic Pesticide Application Plan ants from human sources. Under Section 303(d) of the Clean Water Act, states were required to develop a list of water bodies that do not meet water quality standards; this list is referred to as the “303(d) list.” This list defines low, medium, and high priority pollutants that require immediate attention by State and Federal agencies. Portions of the Estuary have high-priority 303(d) listings for a number of pollutants, including dioxin compounds, furan compounds, PCBs, mercury, copper, nickel, and exotic (plant and animal) species. The most comprehensive information describing water quality in the Estuary comes from the Regional Monitoring Program managed by the San Francisco Estuary Institute (SFEI) and ongoing studies by the Interagency Ecological Program (IEP). In addition, numerous short-term studies that focus on specific sites, resources, or pollutants are conducted on a regular basis by researchers and entities conducting permit-specified monitoring of waste discharges. The primary water quality parameters discussed below are: temperature, salinity, dissolved oxygen (DO), pH, total suspended solids (TSS), turbidity, and pollutants. Temperature. Water temperatures in the Estuary range from approximately 10˚C to 22˚C (50˚F to 71.6˚F). Temperatures are influenced by seasonal solar cycles and variable inputs of river and coastal ocean waters. Temperatures are typically at the higher end of this range along the Estuary margin during daylight hours as the influence of solar energy warms the water. Salinity. The salinity of the Estuary varies spatially and temporally. Along the northern reach the salinity increases from the Delta to the Central Bay. At the mouth of the Sacra- mento River, for example, the mean annual salinity averages slightly less than 2 ppt; in Suisun Bay it averages about 7 ppt; and at the Presidio in Central Bay it averages about 30 ppt. In the South Bay, salinities remain at near-ocean concentrations (32 ppt) during much of the year, except in the vicinity of the San Jose wastewater outfall at Artesian Slough, where salinities are less concentrated. During summer months in dry years, high evaporation rates may cause salinity in South Bay to exceed that of ocean water. Seasonal changes in the salinity distribution within the Estuary are controlled mainly by the exchange of ocean and Estuary water, and by river inflow. River inflow has the greater influence on salinity distribution throughout most of the Estuary because inflow varies widely, while variations in ocean inputs are relatively small. In winter, high flows of freshwater from the Delta lower the salinity throughout the Estuary’s northern reach. High Delta flows also intrude into South Bay, lowering salinity there for extended periods. In contrast, during the summer, when freshwater inflow is low, saline water from the Bay intrudes into the Delta. The inland limit of salinity intrusion varies greatly from year to year. In addition, channel dredging can increase gravitational circulation and enhance salinity intrusion (Nichols and Pamatmat 1988). Dissolved Oxygen. Oxygen concentrations in estuarine waters are increased by the mixing action of wind, waves, and tides; photosynthesis of phytoplankton and other aquatic plants; and high DO in freshwater inflow. DO concentrations are lowered by plant and animal respi- ration, chemical oxidation, and bacterial decomposition of organic matter. The Estuary’s waters are generally well oxygenated, except during summer in the extreme southern end of the South Bay where concentrations are reduced by poor tidal mixing and high water temperature. Typical concentrations of DO range from 9 to 10 milli- grams per liter (mg/l) throughout the Estuary during periods of high river flow, 7 to 9 mg/l during moderate river flow, and 6 to 9 mg/l during the late summer months when

San Francisco Estuary Invasive Spartina Project 5. 2010 Aquatic Pesticide Application Plan flows are the lowest. Unlike the 1950s and 1960s, when inadequately treated sewage and processing plant wastes depleted oxygen in parts of the Bay and Delta, today there are few reports of places in the Estuary where low oxygen concentrations adversely affect beneficial uses. Today, the lowest concentrations in the Estuary are typically observed in the extreme South Bay but, in some instances, DO levels in semi-enclosed embayments such as Richardson Bay can be much lower than in the main water body (SFEI 1994). pH. The pH of the water in San Francisco Bay is relatively constant and typically ranges from 7.8 to 8.21. Total Suspended Solids (TSS) and Turbidity. Turbidity and TSS are generally used as measures of the quantity of suspended particles. The distinction between the two terms lies mainly in the method of measurement. In general, higher TSS results in more turbid water. Regions of maximum suspended solids occur in the North Bay in the null zone2 (generally 50 to 200 mg/l, but as high as 600 mg/l TSS). The specific location of the null zone changes depending upon freshwater discharge from the Delta. TSS levels in the Estuary vary greatly depending on the season, ranging from 200 mg/l in the winter to 50 mg/l in the summer (Nichols and Pamatmat 1988; Buchanan and Schoellhamer 1995). TSS also varies with tidal stage and depth (Buchanan and Schoellhamer 1995). Shallow areas and channels adjacent to shallow areas have the highest suspended sediment concentrations. The Central Bay generally has the lowest TSS concentrations; however, wind-driven wave action and tidal currents, as well as dredged material disposal and sand mining operations cause elevations in suspended solids concentrations throughout the water column. Pollutants. Pollutant loading to San Francisco Bay has long been recognized as one of many factors that has historically stressed aquatic resources. Pollutants enter the aquatic system through atmospheric deposition, runoff from agricultural and urbanized land, and direct discharge of waste to sewers and from industrial activity. The Bay’s sediment can be both a source and a sink for pollutants in the overlying water column. The overall influx of pollutants from the surrounding land and waste discharges can cause increases in sediment pollutant levels. Natural resuspension processes, biological processes, other mechanical disturbances, dredging, and sediment disposal can remo- bilize particulate-bound pollutants. Metals. Ten trace metals in the aquatic system and in waste discharged to the Bay are monitored on a regular basis. Total and dissolved fractions are sampled three times a year at Regional Monitoring Program (RMP) stations throughout the Estuary. Tables 1 and 2 present dissolved and total trace metal concentration ranges in Bay waters during 1998 (SFEI 1998).

1 Water or solutions that are acidic have a pH of less than 7.0, and basic or alkaline water have a pH greater than 7.0. A pH of 7.0 is considered neutral. 2 The null zone is area or region of an estuary where the bottom, high-density and surface, low-density currents have equal and opposite effects. It is defined as the zone where the mean near-bottom speed is zero. The actual location of the null zone migrates in response to changes in river discharge. It is important because it is typically characterized by high concentrations of suspended particulate matter and rapid sediment accumulation.

San Francisco Estuary Invasive Spartina Project 6. 2010 Aquatic Pesticide Application Plan Organic Pollutants. Three general types of trace organic contaminants, polycylic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and pesticides, are measured in San Francisco Bay water on a regular basis. Water column concentrations of dissolved and total PAHs in 1998 ranged from 2.1 to 46 parts per trillion (pptr) and from 20 to 300 pptr, respectively (SFEI 1998). Total PCB concentrations in Bay waters during 1998 ranged from 70 to 7,000 parts per quadrillion (ppq), and were below the U.S. Environmental Protection Agency (U.S. EPA) 4-day (chronic toxicity) water quality criteria (30 pptr) (SFEI 1998). Dissolved PCB concentrations ranged from 12 to 930 ppq. Bay waters also contained measurable concentrations of chlorinated pesticides, including chlordanes and DDTs. Total chlordane concentrations ranged from 21 to 5,700 ppq, while total DDT concentrations ranged from 190 to 9,900 ppq (SFEI 1998). A recent review of historical data from several sources found several previously unidentified organic contaminants in the San Francisco Estuary (SFEI 2002). In this study, p-nonylphenol, a common constituent in detergents and other household products, agricultural surfactants, and many industrial products, was identified in Sacramento and San Joaquin River water (at 19 ng/L and 5 ng/L, respectively), but it was not detected in Estuary water.

Sediment Quality Sediment quality in the Estuary varies greatly according to the physical characteristics of the sediment, proximity to historical waste discharges, the physical and chemical condition of the sediment, and sediment dynamics that change with location and season. Gen-

Table 1. Dissolved Concentrations of Trace Metals in Water Samples (SFEI 1998)

Ag As Cd Cr Cu Hg Ni Pb Se Zn µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L Minimum 0.0002 0.83 0.003 0.09 0.37 0.0003 0.56 0.002 ND 0.07 Maximum 0.006 4.8 0.09 3.8 3.5 0.015 7.2 0.40 6.1 22.5

WQ Criteria 1.9 69 42 1100 5 74 210 90 1-hour WQ Criteria 36 9.3 50 3.1 8.2 8.1 81 4-day

ND – Not detectable at laboratory limits

Table 2. Total Concentrations of Trace Metals in Water Samples (SFEI 1998)

Ag As Cd Cr Cu Hg Ni Pb Se Zn µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L µg/L Minimum 0.002 ND 0.009 0.29 0.42 0.0006 0.63 0.05 ND 0.77 Maximum 0.20 9.4 0.36 101 20 0.73 49.0 15.8 6.8 98.6 WQ Criteria 2.3 69 43 1100 2.1 140 58 1-hour WQ Criteria 36 9.3 50 0.025 7.1 5.6 4-day

ND – Not detectable at laboratory limits

San Francisco Estuary Invasive Spartina Project 7. 2010 Aquatic Pesticide Application Plan erally, the level of sediment contamination at a given location will vary depending on the rate of sediment deposition, which varies with seasons and tides (Luoma et al. 1990). Chemical contaminant dynamics in an estuary are closely associated with the behavior of suspended and deposited sediments. The physical and chemical characteristics of sediments, and the bioavailability and toxicity of sediment-associated chemicals to aquatic organisms, are particularly important in determining their potential impact on environmental quality. While pollutant loading to the Estuary from point and non-point sources has declined dramatically over the past two decades, and surface sediment contamination may be de- clining from historical highs, Bay sediments are still an important source and sink of pollutants. Much of the data documenting concentrations of trace metals and organics in Bay sediments are found in the historical summary of Long and Markel (1992) and in the more recent monitoring efforts by the State’s Bay Protection and Toxic Cleanup Program (BPTCP) (SFBRWQCB 1994) and Regional Monitoring Program (SFEI 1994 and 1998). Concentrations of Metals and Organic Pollutants in Sediments. Mean concentrations of trace metals and organics in sediments vary according to grain size, organic carbon con- tent, and seasonal changes associated with riverine flow, flushing, sediment dynamics, and anthropogenic inputs. Anthropogenic inputs appear to have the greatest effect on sediment levels of copper, silver, cadmium, and zinc, as well as several chlorinated and petroleum hydrocarbons (SFBRWQCB 1994). Ranges in sediment metals and trace organic concentrations during 1998 are listed in Table 3. The table also compares measured concentrations to effects range-low (ER-L) and effects range-median (ER-M) values,

Table 3. Ranges of Trace Pollutants in San Francisco Bay Sediments (SFEI 1998)

SEDIMENT SAMPLES (MG/KG) EFFECTS LEVELS (MG/KG) Minimum Maximum ER-L ER-M Arsenic 3.1 19 8.2 70 Cadmium 0.1 2.1 1.2 9.6 Chromium 63 216 81 370 Copper 8.5 76 34 270 Lead 5.4 65 46.7 218 Mercury 0.03 0.82 0.15 0.71 Nickel 68 228 20.9 51.6 Selenium 0.06 0.52 Silver ND 2.0 1.0 3.7 Zinc 64 256 150 410

TOTAL PAHS 0.033 6.30 4.022 44.792 Total PCBs ND 0.26 0.0227 0.18 Total DDTs No Data 0.00158 0.0461 Total Chlordanes ND 0.0099 0.0005 0.006 Key: Concentrations bolded exceed the Lowest Observable Effects Level (ER-L) Concentrations bolded and underlined exceed the Median Observable Effects Level (ER-M) ND – Not detectable at laboratory limits

San Francisco Estuary Invasive Spartina Project 8. 2010 Aquatic Pesticide Application Plan which are levels that are rarely associated with adverse effects to benthic organisms from exposures to sediment-associated contaminants and levels that are frequently associated with adverse impacts, respectively (Long et al., 1995). For most pollutants, ranges in measured concentrations exceed the respective ER-L values but are below the corresponding ER-M values. The exceptions are mercury, nickel, total PCBs, and total chlordanes, which exceed the ER-M values at one or more locations in the Bay. Some sites within San Francisco Bay, such as Lauritzen Canal, the Port of Oakland near San Lean- dro Bay, and Richmond Harbor, which have been greatly affected by historical contamination, contain sediment pollutant levels which are considerably higher than those measured by the Regional Monitoring Program.

4. DESCRIPTION OF TARGET SPECIES There are one native and four non-native species of cordgrass in the San Francisco Estu- ary. The native species is Pacific cordgrass (S. foliosa), which is not a target of the ISP’s control efforts, and is actually the recipient of conservation by controlling the invasive species). The non-native species are Atlantic smooth cordgrass (S. alterniflora), English cordgrass (S. anglica), Chilean cordgrass (S. densiflora), and salt-meadow cordgrass (S. patens). The non-native Atlantic smooth cordgrass hybridizes with the native Pacific cordgrass, and their offspring (referred to in this APAP as "Atlantic smooth cordgrass hybrids" or "hybrids") are highly invasive and considered non-native. Key aspects of the cordgrass species found in the Estuary are contrasted below. The biological contrasts among these species and their roles in their native habitats help to demonstrate how non- native cordgrasses are likely to alter the Estuary's salt marsh ecosystem. First described is the native Pacific cordgrass, followed by the non-native species.

Native Pacific Cordgrass (Spartina foliosa) The historic range of Pacific cordgrass was confined to estuaries from Point Reyes to Baja California, with large gaps in between; for example, it is historically absent in Mon- terey Bay and Morro Bay. Most of the Pacific cordgrass population exists in San Fran- cisco and San Pablo Bays. Its northern limit is now Bodega Bay, a small and recent natural population. It even more recently established in Tomales Bay, where its population surged following major flood and depositional events of the mid-1990s. Pacific cordgrass is a perennial, salt-tolerant marsh grass, which spreads both sexually, by seed dispersal, and asexually, by long, creeping rhizomes (underground stems, or runners) that propagate small clusters of leafy shoots. Clonal (asexual) growth of rhizomes allows individual plants to form extensive colonies without being pollinated by another plant. A colony thus formed is referred to as a "clone." The slender leafy shoots with seed-heads seldom exceed five feet in height, and most shoots range from about one to three feet tall. The height of the cordgrass plant is related to how well it tolerates submersion in tidewaters, and thus how low in the intertidal zone it can grow. The relatively short stature of Pacific cordgrass corresponds with its limited occupation of lower elevations within the intertidal zone. Pacific cordgrass is genetically very similar to Atlantic smooth cordgrass, but the two species also have significant differences. In size, growth rate, production, and ecological tolerances, Pacific cordgrass is much less robust than Atlantic smooth cordgrass (Smart

San Francisco Estuary Invasive Spartina Project 9. 2010 Aquatic Pesticide Application Plan and Barko 1978, Callaway 1990, Boyer, Callaway and Zedler 2000). Pacific cordgrass grows more luxuriantly in clayey mud than sand, but it naturally grows in substrates ranging from sand and mud to peat. Its leaves and stems wither in fall and are shed in winter, as the clones die back to young shoots and buds near the mud surface. The sparse remains of Pacific cordgrass stands in winter are relatively ineffective in trapping sediment. Pacific cordgrass is generally restricted to a narrow portion of the intertidal zone, between an elevation just above mean sea level and an elevation near the level of the average higher daily tide (mean higher high water, "MHHW"). It tends to fail in competition with plants like pickleweed on the marsh plain, which, in California estuaries, approaches the elevation of the MHHW. This modest range in tidal elevation restricts Pacific cordgrass to the sloping banks of tidal creeks, and the gently sloping upper edges of mudflats where sediment accumulates. This leaves the vast acreages of Pacific tidal flats below mean sea level entirely free of emergent vegetation in natural historic conditions. The vegetated marsh plain (middle to high marsh zone) supports either sparse Pacific cordgrass in lower areas, or none at all. Early experiments with Pacific cordgrass demonstrated that its slender, widely spaced leafy shoots and rhizomes are not as effective at stabilizing sediment compared with At- lantic smooth cordgrass, especially under exposed conditions at the bay's edge (New- combe et al. 1979). Seedlings of Pacific cordgrass are seldom found in established marshes, and appear only intermittently in sheltered upper mudflats. Pacific cordgrass is particularly valued as habitat for the endangered California clapper rail, which spends most of its time foraging for food within, or close to, the protective canopy of cordgrass. Rails can move within Pacific cordgrass stands, and spend most of their time under cover of the cordgrass foliar canopy, usually selecting prey items such as invertebrates inhabiting the cordgrass stands and their edges. In contrast to the clapper rail of southern California tidal marshes, San Francisco Bay clapper rails generally do not construct "floating nests" in Pacific cordgrass; instead, they tend to build nests in gum- plants or pickleweed in the higher marsh.

Atlantic Smooth Cordgrass (Spartina alterniflora) and its Hybrids Smooth cordgrass is the closely related sibling to Pacific cordgrass. In the United States, it occurs along both the Atlantic and Gulf Coasts (Gleason and Cronquist 1991). It is unique among the world's cordgrass species in terms of its growth potential and ecological breadth, and it is the parent species of the other most invasive cordgrass species of hybrid origin, English cordgrass (S. anglica; Adam 1990). The San Francisco Estuary population of Atlantic smooth cordgrass was founded by seed from Maryland in the mid- 1970s, introduced experimentally for one of the first tidal marsh restoration projects on the west coast. We refer to the San Francisco Bay population of smooth cordgrass as At- lantic smooth cordgrass. Atlantic smooth cordgrass is a coarse perennial grass that, like its Pacific relative, spreads both by seed dispersal and by creeping rhizomes that form extensive clonal colonies. In parts of the San Francisco Estuary, the rate of lateral spread by rhizomes averages between 3.3 and 6.6 feet per year, in contrast with native Pacific cordgrass, which spreads only 0.6 to 2.4 feet per year in the same marshes (Josselyn et al. 1993). Similar rates of

San Francisco Estuary Invasive Spartina Project 10. 2010 Aquatic Pesticide Application Plan lateral spread of this species and its hybrids have been recorded more recently in Cogswell Marsh on the Hayward Shoreline (K. Zaremba, M. Taylor, pers. comm.) The size range of Atlantic smooth cordgrass is wide and highly variable, depending on its local genetics and environment. In nutrient-rich, well-drained marsh sediment, such as along tidal creek banks and on newly colonized tidal flats, extensive dense stands can exceed 10 feet in height. On poorly drained marsh flats, its vegetation is typically sparse and short, but its dense root and rhizome network maintains pure stands and effectively binds marsh sediments. The "tall form" and "short form" of this species were so strikingly different that they were long assumed to be distinct varieties, rather than variations based on local environmental conditions. Modern research indicates that factors related to marsh drainage, such as waterlogged soil chemistry (especially accumulation of toxic soil sulfides), excessive salinity, and nutrient deficiency interact to cause the dramatic differences in growth-forms of Atlantic smooth cordgrass (Bradley and Dunn 1989, Mendels- sohn and Seneca 1980, Valiela et al. 1978, Smart and Barko 1978). Genetic variations in height forms of Atlantic smooth cordgrass also has been defined in San Francisco Bay (Daehler et al. 1999) In the salt marshes of the Atlantic coastal plain, Atlantic smooth cordgrass is dominant over most of the intertidal zone. Depending on local tidal range, it can grow to and below mean low water (McKee and Patrick 1988), and it can occupy, and even dominate, the marsh plain and the low marsh. Vast, homogeneous stands of Atlantic smooth cordgrass are the characteristic signature of the Atlantic region's tidal marshes (Dame et al. 2000, Adam 1990, Chapman 1964, 1977). In contrast with Pacific cordgrass, Atlantic smooth cordgrass freely establishes in relatively exposed shorelines with significant wave action, including estuarine sand beaches. It is planted in its native range to stabilize shorelines and to trap and accumulate sediments, and the high density of its tall stems is highly effective at reducing estuarine wave energy (Gleason et al. 1979, Knutson and Woodhouse 1988, Knutson et al. 1990) In other environmental tolerances, Atlantic smooth cordgrass is also highly resilient. It can survive in salinity over 45 parts per thousand (well above ocean salinity), and grow luxuriantly in dilute brackish water. If buried, it can regenerate from up to about one foot of burial by deposited sediment. Atlantic smooth cordgrass, like other low marsh species, can supply air to its roots in oxygen-poor waterlogged mud, using porous air-filled cham- bers linking its foliage to roots and rhizomes. Atlantic smooth cordgrass can also tolerate the severe waterlogging and hypersalinity that develops in poorly drained depressions in the salt marsh, including salt marsh pans. Salt marsh pans are frequent and well- developed features of historic San Francisco Estuary marshes, and important habitat for migratory waterbirds (Goals Project 1999). Along the Hayward shoreline of San Fran- cisco Bay, Atlantic smooth cordgrass has colonized many pre-existing pans, converting them to solid cordgrass marsh. In the San Francisco Estuary, Atlantic smooth cordgrass has displayed many of the ecological traits typical of its performance in its native salt marsh habitat, and some highly novel phenomena as well. Most colonies in the San Francisco Estuary are young, often forming nearly circular, discrete, expanding colonies, which merge into irregular patterns, resembling mold colonies in a petri dish. The edges of the colonies are tall and robust, while the centers often exhibit early symptoms of dieback or "short form" growth habits.

San Francisco Estuary Invasive Spartina Project 11. 2010 Aquatic Pesticide Application Plan The "donut" shape of colonies, in fact, is one of the species' signatures for identification in aerial photographs of San Francisco Bay. This trait is not typical of mature Atlantic salt marshes. In the mild Pacific winters, Atlantic smooth cordgrass shoots tend to retain green leaves and persistent dead leaves through much of the winter. This is an important contrast with native Pacific cordgrass: combined with the invader's much greater stem size and shoot density, year-round dense foliage gives Atlantic smooth cordgrass exceptionally high potential to accumulate and trap estuarine sediment during winter storms or floods. The San Francisco population of Atlantic smooth cordgrass has generated some unusual growth forms with strikingly atypical appearance. The dwarf form develops a profusion of short lateral shoots instead of a tall main stem, forming pure stands with complete ground cover of dense, low turf-like ankle-high vegetation on the marsh plain. The growth rate of the dwarf form is, however, vigorous. The dwarf form is genetic, not environmentally, induced; it occurs in the same local environments that support luxuriant, tall stands of Atlantic smooth cordgrass, often contiguous with the dwarf patches. It has established at multiple locations in San Francisco Bay (Daehler et al. 1999). A comparable dwarf form of its hybrid daughter species, English cordgrass, independently evolved in Britain and New Zealand (Bascand 1970). Hybridization of Atlantic smooth cordgrass with native Pacific cordgrass. Perhaps the most novel and significant phenomenon of the San Francisco population of Atlantic smooth cordgrass is the rapid evolution of an aggressively expanding hybrid swarm formed by cross pollination with the native Pacific cordgrass (Daehler and Strong 1997). The hybrid swarm includes first-generation crosses between Atlantic smooth cordgrass and Pacific cordgrass with both species acting as pollen-parents and seed parents. Be- cause the two species' pollination periods overlap little, first-generation crosses are infrequent. Hybrids, however, have a wide range of flowering times, and act as an effective reproductive bridge between the species. The hybrids produce pollen in much greater abundance (21 times greater) and with higher fertility than the native Pacific cordgrass. Superior hybrid pollen production and fertility so overwhelm populations of Pacific cordgrass ("pollen swamping") that native stands of cordgrass produce mostly hybrid back-cross seeds in the presence of flowering hybrid colonies, and fail to reproduce the species sexually (Ayres et al. 1999, Antilla et al. 2000). This process alone, called hybrid assimilation, can result in the extinction of the invaded species (Levin et al. 1996, Rhy- mer and Simberloff 1996). Genetic analysis has revealed that numerous large populations of presumed Atlantic smooth cordgrass in the Estuary are predominantly hybrids and back-crosses (introgres- sants). The ecologically invasive, dominant traits of Atlantic smooth cordgrass appear to be prevalent in the hybrid swarm. "Pure" Atlantic smooth cordgrass is now a minority in most of the rapidly evolving hybrid swarms, and trends suggest that hybrids will eventually replace both parent species, as the hybrid-origin species English cordgrass did in Britain (see English Cordgrass, below). This recently discovered threat of genetic extinction to a native cordgrass from an alien cordgrass invasion is unique to the San Francisco Estuary. No native cordgrasses existed where Atlantic smooth cordgrass invaded Wash- ington and Oregon estuaries, and the cordgrasses native to Europe are genetically isolated from their hybrids.

San Francisco Estuary Invasive Spartina Project 12. 2010 Aquatic Pesticide Application Plan Atlantic smooth cordgrass, and its hybrids with similar appearance and behavior, are now widely distributed in the Central and South Bay, but they have not yet been detected in the North Bay or Suisun, despite intensive searches. The northern limit of its distribution in 2005 on the east bay is the bay on the western shore of Point Pinole (Giant Marsh), and on the west bay is a clone north of Miller Creek mouth (north of China Camp). The abundance of Atlantic smooth cordgrass and hybrids remains greatest near the point of its original introduction circa 1977 (Pond 3, Hayward Shoreline, Alameda County), and sites of early transplanting (Colma Creek, San Mateo County), early pioneer colonies (Oak- land, San Leandro Bay, Hayward Shoreline), and areas of subsequent transplanting (Cogswell Marsh, Hayward). It now is nearly the exclusive marsh plant species of recently formed or restored tidal marshes along the San Leandro-Hayward shoreline, and this trend is expected to increase. Even as the Bay edge salt marshes and levees are eroding landward through wave action, Atlantic smooth cordgrass marsh is spreading in the opposite direction below the wave-cut marsh cliff. Its distribution becomes patchier south of the Dumbarton Bridge, decreasing in size and frequency to Alviso, where it is still relatively rare. It is well established as scattered, large but discrete colonies in the Dum- barton-Mowry Marsh, Newark, mostly in sloughs and disturbed marsh, or recently colonized mudflats. It is a common or dominant feature in marshes from San Bruno, the San Francisco Airport, south to Foster City, and is scattered in variable frequency along the Redwood City shoreline. The Napa-Sonoma and Petaluma Marshes are currently free from the Atlantic smooth cordgrass invasion, but young colonies have recently been detected in Bolinas Lagoon and Drakes Estero on the Point Reyes peninsula (K. Zaremba, pers. comm. 2005).

English Cordgrass (Spartina anglica) English cordgrass is an aggressive invader of mudflats and salt marshes in Britain, New Zealand, Australia, and the Pacific Northwest, and thrives in cool temperate climates. It originated in Britain as a fertile hybrid derived from introduced Atlantic smooth cordgrass and common cordgrass (S. maritima), a small, slow-growing creeping cordgrass native to European coasts, now greatly reduced in abundance. Within a century after its origin, English cordgrass became the dominant salt marsh grass in Britain (Lee and Partridge 1983, Gray et al. 1990). It is shorter than Atlantic smooth cordgrass and more grayish in appearance, but partly shares other traits of its parent, such as vigorously spreading rhizomes, ability to transform mudflats into vast stands of low marsh vegetation, and ability to dominate and displace associated plant species. It was introduced to the San Francisco Estuary at Creekside Park, Corte Madera, Marin County, along with Chilean cordgrass, in 1976. Unlike Atlantic smooth cordgrass and Chilean cordgrass, this species failed (so far) to disperse from its point of introduction. It may be at or near its southern climatic limit on the Pacific Coast in San Francisco Estuary.

Chilean Cordgrass (Spartina densiflora) Chilean cordgrass (also called dense-flowered cordgrass) is a distinctive cordgrass species native to South America. It has a bunchgrass growth habit, forming tight clumps or tussocks with short creeping rhizomes, and narrow, firm, in-rolled leaves (Spicher 1984), resembling European beachgrass (Ammophila arenaria). It is generally restricted to the middle marsh plain and high marsh zones where pickleweed, saltgrass, jaumea, and other

San Francisco Estuary Invasive Spartina Project 13. 2010 Aquatic Pesticide Application Plan low-growing herbs otherwise prevail. It does not spread into the low marsh where Pacific cordgrass and mudflats naturally dominate the Estuary (Kittleson and Boyd 1997). Chil- ean cordgrass lacks well-developed tissues specialized for transporting air from foliage to roots (Spicher 1984), a feature common to cordgrasses adapted to low marsh environments. Chilean cordgrass, along with other South American coastal species, was probably acci- dentally introduced to Humboldt Bay, California by ship ballast containing seeds from South American ports that traded lumber (Spicher 1984). For most of the 20th Century, Chilean cordgrass was erroneously treated as an "ecotype," or minor geographic variation, of the native Pacific cordgrass, despite the lack of diagnostic traits matching this species. In the late 1970s, the presumed native "Humboldt Bay form" of Pacific cordgrass was deliberately transplanted to salt marsh restoration and landscaping sites at Creekside Park, Corte Madera, Marin County. Within the salt marshes fringing Corte Madera Creek, it has since become a locally dominant component of the middle and high salt marsh vegetation, displacing even robust pickleweed. A second population of Chilean cordgrass spontaneously established across the Bay from Creekside Park in the ancient marsh plain at Point Pinole (Whittell Marsh), Contra Costa County. The Point Pinole population was discovered in the mid-1990s, and has been largely controlled. A single, large, individual clump of Chilean cordgrass established in a very young restored tidal marsh (breached 1995) at the former Salt Pond 2A, Napa Marsh. That pioneer plant was also eradicated.

Salt-Meadow Cordgrass (Spartina patens) Salt-meadow cordgrass is another rhizome-forming creeping cordgrass of Atlantic salt marshes, but unlike Atlantic smooth cordgrass, it has fine stems with narrow, soft, in- rolled leaves, and is intolerant of waterlogged mud. It is naturally confined to the well- drained high salt marsh and relatively moist sandy depressions at or above tidal influence. Two distinctive geographic and ecological types have been recognized, and in the past have been treated as distinct taxonomic varieties. In peaty high salt marshes of the northeastern Atlantic coast, a relatively low form with lax, slender stems forming dense matted turfs with "cowlicks" was once treated as S. patens var. monogyna. These dense salt marsh turfs are often nearly pure stands of salt-meadow cordgrass crowding out most potentially associated species that occupy gaps in the cover caused by winter ice or drifted wracks. In sandy marshes associated with large barrier beaches and wash-over fans from Cape Cod through the Atlantic coastal plain, a coarser, erect type, formerly recognized as S. patens var. juncea, is prevalent. Intermediate forms are common. Between the 1959 publication of A California Flora (Munz and Keck 1959) and its 1970 supplement, salt-meadow cordgrass was reported in Southampton Bay, Benicia, Solano County. The time and mode of introduction is unclear. Salt-meadow cordgrass at South- ampton occupies large, discrete patches in pure and exceptionally thick stands compared with its native marshes. The patches are distributed close to tidal sloughs, a pattern suggesting local transport by currents. One large stand is spreading into a high marsh site (pickleweed-saltgrass vegetation) that supports a population of an endangered annual plant, soft bird's-beak (Cordylanthus mollis ssp. mollis). The Southampton Bay cordgrass

San Francisco Estuary Invasive Spartina Project 14. 2010 Aquatic Pesticide Application Plan population appears to match the type description of "variety monogyna," the fine-stem type of northeastern Atlantic marshes.

5. CONTROL TOLERANCES As previously mentioned, the non-native Spartina invasion of the San Francisco Estuary is especially threatening to native marsh systems because of hybridization of Atlantic smooth cordgrass and the native Pacific cordgrass. This ability to hybridize, and the current expansion rates of the population of hybrid forms throughout the Estuary, defines the need for a zero tolerance threshold on invasive Spartina in San Francisco Bay. The Inva- sive Spartina Project is a regionally coordinated eradication effort that will ultimately be successful only if all infestations are effectively controlled and monitored to eradication. A single small, expanding clone of hybrid Spartina within an otherwise native S. foliosa matrix has the capability of ‘swamping’ S. foliosa flowers with hybrid pollen, effectively converting the native stand into a hybrid-producing population. Within a couple of growing seasons, the majority of new seedlings establishing in the area will be of hybrid origin, resulting in the eventual extirpation of the native S. foliosa from the stand. Repeated throughout the Estuary on various scales, this progression threatens the population stabil- ity of native pacific cordgrass stands. Therefore, where hybrid forms of Spartina are identified, efforts must be directed at removing all of the plants in the area. There is no acceptable level of hybrid presence in an otherwise native marsh, as the inevitable result of even a small amount of hybrid presence will be the relatively rapid conversion of the marsh to a non-native stand capable of infesting adjacent marshlands. Spartina densiflora has also been shown to have very rapid expansion rates in the Estu- ary. In addition, this plant invaded Humboldt Bay, CA to such a degree that approximately 80% of the mudflats and marsh lands have been dominated by it. We are fortunate in the San Francisco Estuary to have only isolated populations of S. densiflora at this point because of the efforts of the ISP and its partners, so as in the case with the more prevalent S. alterniflora hybrids, there is a need for zero tolerance and an excellent oppor- tunity for the eradication of this species before it becomes widespread. For the other two invasive Spartina species in the Estuary, S. anglica and S. patens, each only inhabits a single infestation site and in a relatively small overall area. However, S. anglica is know to be the most invasive species of Spartina in the world, itself a fertile hybrid that has dominated introduction sites on several continents. With these types of threats, the fertile environment of the Estuary, and the current size of the infestations, a zero tolerance policy is appropriate. In addition, with a couple effective seasons of treatment, each of these species will be eradicated from the Estuary.

6. DESCRIPTION OF HERBICIDE Herbicides have proven highly effective in controlling populations of cordgrasses (Spartina spp.). The aquatic formulation of imazapyr (Habitat®) was registered for use in the State of California on August 30, 2005. Previously, the Spartina Control Program used aquatic glyphosate-based herbicides (Aquamaster®, Rodeo®), which were the only

San Francisco Estuary Invasive Spartina Project 15. 2010 Aquatic Pesticide Application Plan herbicide formulations that could be used in sensitive estuarine systems and were designed to work on grasses/monocots. All ISP partners have since switched to imazapyr, although glyphosate will also be used at a single sub-site with special circumstances, and may be combined with imazapyr (according to the product labels) as a “brown-down indicator” to allow for more rapid detection of missed or skipped areas. Since imazapyr is such a slow-acting herbicide, it is difficult to know if the entire infestation at a site has been effectively treated until the following spring. Glyphosate treatment results in more noticeable yellowing/browning of the treated plants within a couple weeks. The use of a brown-down indicator would make any green, untreated plants stand out, and a follow-up spot treatment could be applied to these plants without losing a year of control. There are a number of qualities that make imazapyr the ISP’s preferred choice over the previous alternative, glyphosate. Glyphosate tends to be strongly adsorbed to sediment and salt particles accumulated on the Spartina, rendering the herbicide inactive. It is common for the tides to deposit abundant sediment from the turbid San Francisco Bay onto the invasive Spartina in the adjacent salt marshes. Glyphosate also requires significantly longer dry times to fully penetrate the cuticle of the plant and begin translocation. Imazapyr does not have these issues that can reduce its efficacy. In addition, imazapyr is applied at lower concentrations and the applicator does not need to “spray to wet” as with glyphosate. This greatly reduces the amount of herbicide that will enter the environment as a result of the Spartina Control Program’s efforts, and allows for low volume applications such as aerial helicopter work. This herbicide delivery system is essential to the ISP’s initial efforts because of the vast acreage of Spartina infesting hazardous and difficult to access marshes. Aquatic herbicide formulations such as those used by ISP must be combined with a suitable surfactant to facilitate uptake and translocation of the herbicide down into the rhizomes. A harmless, inert marker dye or colorant is often added to the tank mix to assist the applicator at achieving full coverage while not over-applying to any areas. The following discussion addresses imazapyr, glyphosate, breakdown products, and typical surfactants and colorants. Detailed descriptions of the chemical properties, degradation rates, environmental fate, and toxicity of imazapyr, glyphosate, and all of the aquatic surfactants evaluated for the Spartina Control Program are provided in Appendix 1. Imazapyr. Habitat® or Polaris™ are solutions of 28.7% isopropylamine salt of imazapyr in water, equivalent to 22.6% imazapyr acid equivalents (a.e.) or 2 lbs. acid per gallon, and contain a small amount of an acidifier. Because Habitat® is purportedly the same formulation as Arsenal® and this product contains acetic acid, the acidifier in Habitat® is likely also acetic acid (Leson & Associates 2005.) No information has been found in the published literature on manufacturing impurities associated with imazapyr. Because vir- tually no chemical synthesis yields a totally pure product, technical grade imazapyr most likely contains some impurities. However, to some extent, concern for impurities in technical grade imazapyr is reduced by the fact that most existing toxicity studies on imazapyr were conducted with the technical grade product and encompass the toxic potential of the impurities (SERA 2004). A generic version of this aquatic imazapyr formulation is now available from NuFarm under the product name Polaris AQ™. Imazapyr inhibits an enzyme in the biosynthesis of the three branched-chain aliphatic amino acids valine, leucine, and isoleucine. Because animals do not synthesize branched-

San Francisco Estuary Invasive Spartina Project 16. 2010 Aquatic Pesticide Application Plan chained aliphatic amino acids but obtain them from eating plants and other animals, the engineered mechanism for plant toxicity, i.e. the interruption of protein synthesis due to a deficiency of the amino acids valine, leucine, and isoleucine, is not generally relevant to birds, mammals, fish or invertebrates. Any toxicity to these receptors occurs through different mechanisms (Entrix 2003). Imazapyr is relatively slow acting taking several weeks for the plants to show lethal effects. However, plants cease growth within 24 hours of a successful application (J. Smith, pers. comm. 2006). On Spartina, it takes 2-4 weeks after treatment to see visible effects such as yellowing of the leaves, and complete plant death can take several months. In the San Francisco Estuary, with the relatively late season applications on invasive Spartina (mainly because of endangered species issues that effect access to the marshes), the treated plants may not reveal much of a response before natural senescence, but will simply not emerge in the spring of the following year if fully impacted. Glyphosate. Aquamaster® and Rodeo® are aqueous solutions containing 53.8% glyphosate in its isopropylamine salt form or 4 lbs. acid per gallon, and contain no inert ingredients other than water. The primary decomposition product of glyphosate is aminomethylphosphonic acid (AMPA), and the commercial product contains an impurity, 2,4-nitrosoglyphosate (NNG). The potential effects of AMPA and NNG are encompassed by the available toxicity data on glyphosate and glyphosate formulations (SERA 1996). Glyphosate inhibits an enzyme needed to synthesize an intermediate product in the biosynthesis of the aromatic amino acids, essential for protein synthesis and to produce many secondary plant products such as growth promoters, growth inhibitors, phenolics, and lignin. Animals do not synthesize these aromatic amino acids and glyphosate therefore has low toxicity to these receptors. (Schuette 1998.) In general, glyphosate herbicides are somewhat faster acting than imazapyr herbicides. On Spartina, complete brown-down occurs within 7 to 21 days. (K. Patten, pers. comm. 2004). Surfactants. For most foliar applications of aquatic herbicide formulations, adjuvants must be added to spray solutions to improve the performance and minimize variation of herbicide efficacy. Surfactants are designed to improve the spreading, dispers- ing/emulsifying, sticking, absorbing, and/or pest-penetrating properties of the spray mixture (Tu et al. 2001). The pure herbicide formulation mixed with water will stand as a droplet on the waxy leaf surface and the small area of contact therefore provides little potential for uptake of the active ingredient into the foliage. Water droplets containing a surfactant will spread in a thin layer over a waxy leaf surface and improve herbicide uptake by maximizing herbicide distribution and forcing the fluid into the plant. As men- ® tioned above, both Habitat and Polaris™, as well as the glyphosate herbicides Aquamas- ter® and Rodeo®, require the addition of surfactants for post-emergent applications such as the control of invasive Spartina. Imazapyr. The Habitat® specimen label recommends a variety of different spray adjuvants for use on post-emergent vegetation. For non-ionic surfactants the label recommends a rate of 0.25% v/v or higher, preferably of a surfactant with a hydrophilic to lipophilic ratio between 12 and 17 and with at least 70% surfactant in the formulated product (BASF 2003). Alternately, the label recommends the use of methylated seed oils or vegetable oil concentrates at the rate of 1.5 to 2 pints per acre. For spray volumes greater than 30 gallons per acre, the surfactant should be mixed at a rate of 1%. The label

San Francisco Estuary Invasive Spartina Project 17. 2010 Aquatic Pesticide Application Plan further indicates that these oils may aid in imazapyr deposition and uptake by the plants under moisture or temperature stress. Silicone-based surfactants, which may reduce the surface tension of the spray droplet, allowing greater spreading on the leaf surface as compared to conventional non-ionic surfactants, are also recommended. However, the manufacturer points out that some silicone-based surfactants may dry too quickly, limit- ing herbicide uptake (BASF 2004). One study from Washington State concluded that the esterified seed oil surfactant tested, Competitor®, performed better than the other surfactants tested, i.e. Agri-Dex®, a crop oil-based surfactant, and R-11®, a non-ionic surfactant. This finding is also supported by Patten (2002) in which the author recommended using a methylated seed oil surfactant for aerial applications and for unfavorable conditions such as less than 6 hours of drying time or moist leaves. The experience of the ISP from 2005-2007 has shown that a lecithin product, Liberate®, has also been highly effective with imazapyr. In addition, this product acts as a drift retardant which may help in helicopter treatments as well as other broadcast applications to ensure full coverage on the target Spartina and minimize drift onto non- target marsh plants. Glyphosate. The Aquamaster® and Rodeo® specimen labels recommend the use of a non-ionic surfactant containing at least 50% active ingredient at a rate of 2 or more quarts per 100 gallons of tank mix (0.5% v/v). With glyphosate it is also important to balance the pH of the tank water to ensure effectiveness, and some adjuvants are designed with this purpose in mind, namely LI-700®. Not all surfactants provide the same effectiveness and surfactant costs vary widely. In general, non-ionic surfactants and crop oil concentrates are the least expensive of the surfactant classes, followed by esterified seed oils and organo-silicates. (Miller & Westra 2004.) The ISP identified a number of potential surfactants for use with imazapyr and glyphosate. They include the non-ionic surfactants Liberate® and LI-700®, the crop-oil concentrate Agri-Dex®, the esterified seed oil Competitor®, and the organo-silicones Dyne-Amic® and Kinetic®. Based on the efficacy experienced by the ISP from 2004- 2007, and their superior relative toxicities, the ISP expects to continue to exclusively use Competitor® and Liberate® in the 2008 Spartina Control Program. LI-700® may be used to balance the pH of the tank if/when glyphosate is employed. Cygnet Plus® was evaluated and originally included in the ISP list of surfactant choices for 2005, but it was shown to be ineffective with imazapyr and has been removed from the list available to the ISP partners. Competitor (Wilbur-Ellis Company) is a modified vegetable oil containing a non-ionic emulsifier system. The ingredients include ethyl oleate, sorbitan alkyl polyethoxylate ester, and dialkyl polyoxy-ethylene glycol. Toxicity studies classified this surfactant as a toxicity category of 3-4 (CAUTION). Liberate (Loveland Industries, Inc.) is a non-ionic, low foam penetrating adjuvant. Its active ingredients are lecithin (phosphatidylcholine, which is a naturally occurring lipid derived from soybeans that biodegrades readily), methyl esters of fatty acids, and alcohol ethoxylate. In a 1% solution, the pH is an almost neutral 6.8. Toxicity studies classified this surfactant as a toxicity category of 3-4 (CAUTION). It improves deposition and re- tards drift by producing a more uniform spray pattern.

San Francisco Estuary Invasive Spartina Project 18. 2010 Aquatic Pesticide Application Plan Dyne-Amic (Helena Chemical Company) is a proprietary blend of non-ionic organosilicone surfactants and a methylated vegetable oil. Toxicity studies classified this surfactant as a toxicity category of 3-4 (CAUTION). Kinetic (Helena Chemical Company) is a non-ionic wetting agent that allows for the rapid spreading and absorption of herbicide sprays into the target vegetation, and is especially effective with water-based herbicide formulations. Its active ingredients include organosilicone and polyoxypropylene-polyoxyethylene copolymer. Toxicity studies classified this surfactant as a toxicity category of 3-4 (CAUTION). Agri-Dex (Helena Chemical Company) is a non-ionic surfactant consisting of a paraffin base petroleum oil, polyol fatty acid esters, and polyethoxylated derivatives of the fatty acid esters. Toxicity studies classified this surfactant as a toxicity category of 3-4 (CAUTION).3. Biodegradation of this adjuvant is presumed to be rapid. LI-700 (Loveland Industries, Inc.) contains phosphatidylcholine (lecithin), which is a naturally occurring lipid that biodegrades readily. It also contains methylacetic acid and alkyl polyoxyethylene ether. Toxicity studies classified this surfactant as a toxicity category of 1 (DANGER) because of corrosive properties to the skin and/or eyes. Biodegradation of this adjuvant is presumed to be rapid because of the natural lecithin ingredients. Colorant. There are several colorants suitable for use in the marsh environment, all of which are similar in composition and performance. Blazon Spray Pattern Indicator (Mil- liken Chemical), a typical colorant, is a water-soluble polymeric product. As with most colorant products, the active ingredients in Blazon are proprietary; the Material Safety Data Sheet (MSDS) indicates that it is non-hazardous and non-toxic. The product information sheet reports that the product is non-staining to the skin or clothing. A literature survey on the toxicity of color indicators done for the U.S. Department of Agriculture reports “most commercial indicators are blue … and most often a form of Acid Blue 9…” (McClintock 1997 and Zullig 1997 cited in SERA 1997b). Acid Blue 9 is a disodium salt classed chemically as a triphenelmethane color (SERA 1997b). The Cosmetic, Toiletry, and Fragrance Association (CTFA) name for certified batches of Acid Blue 9 is FD&C blue No. 1. Herbicide application. Impacts to water quality from herbicide application depend on environmental fate, degradation rates of active agents and decomposition products of the herbicides. The primary route by which herbicide solution may contact water is by overspray directly onto the water surface or onto areas that will be covered by water on the next tide. Herbicide may also be washed off plants by tidal inundation or precipitation. Imazapyr and glyphosate solutions will be applied as sprays to Spartina foliage for control of this invasive plant. Spray mixtures may be dispersed from manually transported tanks (backpack sprayers) or spray equipment mounted on trucks, amphibious tracked vehicles, boats, or helicopters. Application rates will be consistent with the product labels (Table 4). Applications from backpack sprayers or conventional spray truck entails workers walking through the marsh and applying herbicide directly to target plants, with limited overspray

3 Toxicity categories are determined by the U.S. EPA for human health affects. See http://www.epa.gov/oppfead1/labeling/lrm/chap-08.htm for more information on pesticide label requirements.

San Francisco Estuary Invasive Spartina Project 19. 2010 Aquatic Pesticide Application Plan to surrounding plants or water surfaces. Spot application from amphibious tracked vehicles or boats would entail vehicles moving through the marsh or adjacent waterway applying herbicide with hand-held equipment to target vegetation with limited overspray. Aerial ap-plication is conducted by helicopter (the imazapyr label does not allow for fixed-wing air-craft to be used) with either a boom sprayer (a horizontal pipe with spray nozzles along its length, mounted to the bottom of the helicopter) or a spray ball (a hollow ball with nozzles suspended from the bottom of the helicopter). Broadcast aerial application involving a boom sprayer would result in a wider dispersion of herbicides, with greater potential for overspray onto non-target areas or the water surface. However, for aerial treatment on smaller patches of invasive Spartina, the boom may be shortened up and the apparatus turned on only when over the target vegetation. Aerial application will be used primarily at large areas of dense cordgrass infestations, particularly in locations where little native cordgrass and other non-target plants are nearby. Applications from backpack sprayers or conventional spray truck entails workers walking through the marsh and applying herbicide directly to target plants, with limited overspray to surrounding plants or water surfaces. Spot application from amphibious tracked vehicles or boats would entail vehicles moving through the marsh or adjacent waterway applying herbicide with hand-held equipment to target vegetation with limited overspray. Aerial application is conducted by helicopter (the imazapyr label does not allow for fixed-wing air- craft to be used) with either a boom sprayer (a horizontal pipe with spray nozzles along its length, mounted to the bottom of the helicopter) or a spray ball (a hollow ball with nozzles suspended from the bottom of the helicopter). Broadcast aerial application involving a boom sprayer would result in a wider dispersion of herbicides, with greater potential for overspray onto non-target areas or the water surface. However, for aerial treatment on smaller patches of invasive Spartina, the boom may be shortened up and the apparatus turned on only when over the target vegetation. Aerial application will be used primarily at large areas of dense cordgrass infestations, particularly in locations where little native cordgrass and other non-target plants are nearby.

Environmental Fate of Herbicides Herbicide mixtures may be indirectly discharged to surface waters by tidal action or rainfall that rinses the herbicide solution from the plants. Rainfall is unlikely to occur during the planned application season (summer in the San Francisco Bay region), and herbicide applications would be postponed if rainfall were predicted, but tidal inundation is inevitable on a regular cycle. Applications to invasive Spartina along slough or creek banks or at the Bayfront on fringe marshes and mudflats will result in a small percentage of the herbicide directly entering the water column. Food-web exposures become significant only with chemicals that have a tendency to bio- accumulate or biomagnify. The adverse effects associated with bioaccumulative chemicals relate to their propensity to transfer through the food web and accumulate preferen- tially in adipose or organ tissue. Basic routes for organism exposure to bioaccumulative compounds are the transport of dissolved contaminants in water across biological membranes, and ingestion of contaminated food or sediment particles with subsequent transport across the gut. For upper-trophic-level species, ingestion of contaminated prey is the predominant route of exposure, especially for hydrophobic chemicals.

San Francisco Estuary Invasive Spartina Project 20. 2010 Aquatic Pesticide Application Plan Table 4a: Imazapyr herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Estu- ary (Leson & Associates 2005.)

Application Method Spray Volume Formulation Active Ingredient1 Surfactant2 Colorant 0.25% v/v NIS with ≥70% a.i.; High volume hand- 0.52-0.75% solution 100 gal/acre 1-1.5 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 3 qt/100 gal held sprayer 4-6 pints/100 gal SBS according to label 0.25% v/v NIS with ≥70% a.i.; Low-volume directed 0.75-1.5% solution 20 gal/acre 0.3-0.6 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 3 qt/100 gal sprayer 1.2-2.4 pints/20 gal SBS according to label 0.25% v/v NIS with ≥70% a.i.; Broadcast sprayer/ 2.5-7.5% solution 10-30 gal/acre 0.5-1.5 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 0.5-1.5 qt/acre Aerial application 6 pints/10-30 gal SBS according to label

1 Active ingredient in Habitat® is imazapyr isopropylamine salt; values expressed as imazapyr acid equivalent 2 NIS = non-ionic surfactant; MSO = methylated seed oil; ESO = esterified seed oil; VOC = vegetable oil concentrate, SBS = silicone-based surfactant, %v/v = percentage based on volume by volume

Table 4b: Glyphosate herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Es- tuary (Leson & Associates 2005.)

Application Method Spray Volume Formulation Active Ingredient1 Surfactant2* Colorant

High volume hand- 1-2% solution 100 gal/acre 4-8 lb a.e./acre ≥0.5% v/v NIS with ≥50% a.i. 3 qt/100 gal held sprayer 1-2 gal/100 gal

Low-volume di- 1-8% solution 25-200 gal/acre 1.35-10.8 lbs a.e./acre ≥0.5% v/v NIS with ≥50% a.i. 3 qt/100 gal rected sprayer 1-8 gal/100 gal

Broadcast sprayer/ 7-40 gal/acre/ 4.5-7.5 pints/acre 2.25-3.75 lb a.e./acre ≥0.5% v/v NIS with ≥50% a.i. 0.5-1.5 qt/acre Aerial application 7-20 gal/acre

1 ® ® The active ingredient in Rodeo and Aquamaster is glyphosate isopropylamine salt; values are expressed as glyphosate acid equivalent 2 NIS = non-ionic surfactant, %v/v = percentage based on volume by volume

San Francisco Estuary Invasive Spartina Project 21. 2010 Aquatic Pesticide Application Plan U.S. EPA’s Hazardous Waste Identification Rule (USEPA 1999) identifies compounds that are recognized as having a low, medium or high potential for bioaccumulation. For bioaccumulation in aquatic systems, rankings were determined using bioaccumulation factors in fish, which are indicated in laboratory tests as having low octanol-water parti- tioning coefficient (or log Kow)) values for organic compounds. Bioaccumulation potential is defined as follows: Bioaccumulation Bioaccumulation Factor potential (BAF) log Kow

High BAF >= 10,000 log Kow >= 4.0

Medium 10,000 > BAF >= 100 4.0 > log Kow >= 2.0

Low BAF < 100 log Kow < 2.0 Imazapyr. Under typical environmental conditions of pH 5-9, imazapyr is ionized and is therefore highly soluble in water. Because of its high solubility, imazapyr has an inher- ently low sorption potential with a low soil organic carbon sorption coefficient (Koc) of 8.81 (log Koc), suggesting very high mobility in soil and little adsorption to suspended solids and sediment. Its octanol/water partition coefficient (Kow) has been reported at 0.22 (log Kow), reflecting its high solubility in water and low solubility in lipids, and hence low propensity to bioconcentrate. A low bioconcentration factor (BCF) of 3 was calculated for imazapyr, which suggests a low potential for bioconcentration in aquatic organisms (Leson & Associates 2005). Imazapyr is relatively mobile in soils because it only weakly adsorbs to soils and sediments. Adsorption increases with decreasing pH. Above a pH of 5, imazapyr is ionized and does not adsorb to soil. Aerobic degradation in soils occurs primarily by very slow microbial metabolism with quinoline as the main metabolite (Entrix 2003). Conditions in sediments differ substantially from those in soils, both in terms of the regular exchange of waters within the sediment pore water and over it, and in the degree of oxygenation in sediments that affect microbial metabolism. Because the pH of sediment surfaces and sediment pore water in intertidal mudflats is above neutral (pH >7), imazapyr will be entirely in its ionized form. Thus, adsorption to sediments is expected to be minimal. (Entrix 2003.) Microbial metabolism in sediments has been determined to be insignificant. One study determined the half-life of imazapyr in the pore water of aerobic sediment at 17 months. The degradation of imazapyr when applied directly to water largely mimics the pathway by which the herbicide would be mobilized at high tide after application to Spartina during low tide. Residual imazapyr on the plants that have not completely dried or did not get absorbed by the plants will be inundated by the incoming tide and presumably solubi- lized. Aquatic degradation studies under laboratory conditions demonstrated rapid initial photolysis of imazapyr with reported half-lives ranging from 3 to 5 days (SERA 2004). The two primary photodegradation products were rapidly degraded with half-lives less than or equal to 3 days and eventual mineralization to carbon dioxide (Entrix 2003). Degradation rates in turbid and sediment-laden waters, common to estuarine environments, are expected to be lower than those determined under laboratory conditions. In controlled field dissipation studies in two freshwater pond systems with application of 1.5 lb imazapyr a.e./acre, imazapyr rapidly dissipated from the water with first-order half-

San Francisco Estuary Invasive Spartina Project 22. 2010 Aquatic Pesticide Application Plan lives of 1.9 days and 12.8 days. No detectable residues of imazapyr were found in the water and sediment after 14 and 59 days, respectively. (Entrix 2003.) The ISP’s NPDES water quality monitoring at treatment sites over the past several years has found a standard reduction in imazapyr in the adjacent surface water of more than 95% one-week post- treatment over the amount present immediately after the application. In estuarine systems, dilution of imazapyr in the incoming tide will contribute to its rapid dissipation and removal from the area where it has been applied. Studies in estuaries in Washington State examined the fate of imazapyr applied at a standard rate of 1.5 lb imazapyr a.e./acre directly to sediment. The study design was conservative because imazapyr was applied to bare mudflats with no algal or emergent vegetation intercepting the herbicide. The study measured immediate maximum concentrations of imazapyr in intertidal waters and sediment less than 3 hours after application and short-term concentrations between 24 and 72 hours after application. Sediment samples collected 3 hours after application were retrieved immediately after the first tidal wash over the area. Maximum concentrations in water and sediment were detected at 3.4 mg/L and 5.4 mg/kg, respectively. Measurable concentrations of imazapyr declined exponentially in both water and sediment, approaching the zero-asymptote at 40 and 400 hours with half-lives of <0.5 and 1.6 days, respectively. Water collected 20 and 200 feet outside the spray zone with the first incoming tide was 99% lower than the maximum water concentration at the edge of the spray zone (Leson & Associates 2005). Application of the same amount of herbicide to a stand of 5.5-foot tall Spartina resulted in a 75% reduction in concentrations in sediment through interception by the canopy (Patten 2003). In sum, this research suggests that imazapyr quickly dissipates in estuarine environments. In addition, the same researcher observed that other vegetation rapidly colonizes the plots treated with imazapyr after the Spartina plants have died, which supports the conclusion of very low persistence of imazapyr in estuarine environments. The ISP has observed this phenomenon of rapid native plant colonization of treated areas at many sites around San Francisco Bay since 2005, usually involving either annual pickleweed (Salicornia europaea), perennial pickleweed (Sarcocornia pacifica), or Jaumea carnosa. Glyphosate. Under typical environmental conditions of pH 5-9, glyphosate is ionized. Glyphosate and its salts are readily soluble in water with a solubility of about 12,000 mg/L. Its interactions with soil and sediment are primarily ionic, rather than hydrophobic and pH dependent. Laboratory and field studies indicate that glyphosate is strongly and reversibly adsorbed by soil, sediment, and suspended sediment. Glyphosate is inactivated through soil adsorption. Because glyphosate adheres strongly to particles, it does not readily leach to waters (Sprankle et al., 1977 cited in Albertson, 1998), and potential movement of glyphosate to groundwater is unlikely. Due to its negligible vapor pressure (7.5×10-8 mmHg) and its ionic state in water, glyphosate is not expected to volatilize from water or soil. Glyphosate’s Kow has been reported at 0.00033, indicating its high solubility in water, low solubility in lipids, and thus low potential to bioconcentrate. All reported bioaccumulation factor values for glyphosate in aquatic organisms are well below 100 (Ebasco 1993; Heyden 1991; Wang et al. 1994). The highest bioaccumulation factor of 65.5 was reported for tilapia (a species of fish) in fresh water (Wang et al. 1994). Other studies report much lower bioaccumulation factors in the range of 0.03 to 1.6 for fish (Ebasco 1993). Most studies report rapid elimination and depuration from aquatic organisms after exposure stops (Ebasco 1993). Therefore, bioaccumulation of

San Francisco Estuary Invasive Spartina Project 23. 2010 Aquatic Pesticide Application Plan glyphosate is considered to be low and food-web transfer is not considered to be a significant exposure route. Soil studies have determined glyphosate half-lives ranging from 3 to 130 days. The soil field dissipation half-life averaged 44 to 60 days. (Leson & Associates 2005.) In the soil environment, glyphosate is resistant to chemical degradation, is stable to sunlight, is relatively non-leachable, and has a low tendency to runoff (except as adsorbed to colloidal matter). It is relatively immobile in most soil environments as a result of its strong adsorption to soil particles. Glyphosate is rapidly and strongly adsorbed to sediment, which appears to be the major sink for glyphosate in aquatic systems. Like in soils, the herbicide is inactivated and biodegraded by microorganisms. Several studies indicate that glyphosate is stable in water at pH ranging from 3 to 6. The photolytic half-life of glyphosate in deionized water exposed outdoors to sunlight was approximately 5 weeks at 100 ppm and 3 weeks at 2000 ppm. Glyphosate shows little propensity toward hydrolytic decomposition. Its hydrolysis half-life is greater than 35 days. It is also stable to photodegradation under visible light but photolyzes when exposed to UV radiation. Glyphosate’s loss from water occurs mainly through sediment adsorption and microbial degradation. The rate of microbial degradation in water is generally slower because there are fewer microorganisms in water than in most soils. Studies conducted in a forest ecosystem found that glyphosate dissipated rapidly from surface water ponds high in suspended sediment, with first order half-lives ranging from 1.5 to 11.2 days. In streams, residues were undetectable within 3 to 14 days. Other studies using water from natural sources determined glyphosate’s half-life ranging from 35 to 63 days. (Leson & Associates 2005.) For all aquatic systems, sediment appears to be the major sink for glyphosate residue. A review of the literature on glyphosate dissipation applied under estuarine conditions suggests that 24 to 48 hours after applications, glyphosate concentrations in water were reduced by more than 60-fold. Energetic tidal cycles and tidal currents effectively disperse bound (adsorbed) glyphosate and surfactants and dilute them in microbially active suspended sediment. Studies of the fate of glyphosate and surfactants applied in tidal marshes and mudflats have reported that concentrations of both substances dropped below detection levels as soon as two tidal cycles (one day) to seven days (Kroll 1991, Paveglio et al. 1996) after application. The initial tidal submergence of sprayed surfaces disperses a large fraction of applied glyphosate and surfactant. Research conducted for the California Department of Food and Agriculture (Trumbo 2002) studied the environmental fate and aquatic toxicity of Rodeo and R-11 in three locations, including a Sacramento-San Joaquin Delta slough, a riverine area, and a no- outlet pond. This study measured glyphosate, aminomethylphosphonic acid (AMPA; glyphosate’s primary metabolite), nonylphenol ethoxylate, and nonylphenol at treated sites one hour, two days, and eight days after application. The study also tested for toxicity using 96-hour toxicity tests with the fish species fathead minnow Pimephales promelas. The study found that concentrations of the tested constituents at slough and river sites (with moving water) was below detectible levels for all tests, and that there was no significant mortality of test fishes. The pond site, however, showed detectable residues of glyphosate, nonylphenol ethoxylate, and nonylphenol at one hour and two days after treatment, but all constituents were below detection limits by day eight. The one-hour pond samples experienced 30% mortality of test fishes, which, because of the relatively

San Francisco Estuary Invasive Spartina Project 24. 2010 Aquatic Pesticide Application Plan low concentrations of glyphosate (which is known to be non-toxic at the detected level), was attributed to effects caused by nonylphenol ethoxylate and nonylphenol. The two- and eight-day tests showed no significant mortality to test fishes. Patten (2002) compiled data on the fate of glyphosate in water and sediment following applications in estuarine environments. Data are presented as geometric means for immediate maximum concentration (<3hrs after application) and short-term concentration (between 24 hrs and 48 hrs after application). For use rates between 8 and 16 kg/ha (7-15 lbs/acre), the immediate maximum geometric mean glyphosate concentrations were 0.174 mg/L (174 μg/L) in water and 2 mg/kg in sediment. The short-term geometric mean glyphosate concentrations were 0.003 mg/L (3 μg/L) in water and 1.9 mg/kg in sediment. These independent lines of research in the fate of imazapyr or glyphosate combined with a surfactant in tidal (and other) habitats suggest that potential impacts to water quality and beneficial uses of waters of the State caused by spraying these herbicide mixtures in intertidal environments are likely be small and temporary. Therefore, controlled applications (i.e., following label instructions) of registered herbicides are not expected to degrade water quality, except for limited temporal and spatial extent. In summary, the use of imazapyr or glyphosate combined with a surfactant to treat infestations of non-native cordgrass would result in less than significant impacts on water quality due to the rapid degradation rate and controlled application of herbicides only on target plants. Since application of herbicides would take place during low tide and low wind conditions, the herbicide would likely be absorbed by plants for a minimum of several hours (up to several weeks in high marsh) following application resulting in less than significant quantities of imazapyr, glyphosate or surfactants entering the water.

Potential Biological and Ecological Effects The known properties of the herbicides, potential methods of application, and the ecological characteristics of the Estuary were evaluated to develop a conceptual model (Fig- ure 2) and identify likely receptors and exposure pathways. This model includes identification of primary and secondary herbicide sources, release mechanisms, exposure media, exposure routes, and potential ecological receptors. For effects to occur, a receptor and a complete exposure pathway must be present. An exposure pathway is only considered complete when all four of the following elements are present: project- related source of a chemical, a mechanism of release of the chemical from the source to the environment, a mechanism of transport of the chemical to the ecological receptor, and a route by which the receptor is exposed to the chemical. The exposure routes associated with the complete pathways include direct contact with the herbicide mixture during and immediately after application, ingestion of contaminated surface water and sediments, direct contact with contaminated surface water and sediments, and food-web exposure. The conceptual model (Figure 2) illustrates the links between sources, release and transport mechanisms, affected media, exposure routes, and potentially exposed ecological receptors. Although several complete exposure pathways may exist, not all pathways are comparable in magnitude or significance. The significance of a pathway as a mode of exposure depends on the identity and nature of the chemicals involved and the magnitude of the likely exposure dose. For birds and mammals, ingestion is generally the most significant exposure pathway. Dermal contact is ex

San Francisco Estuary Invasive Spartina Project 25. 2010 Aquatic Pesticide Application Plan Herbicide Mixture in Tanks and PRIMARY Equipment SOURCE

RELEASE Accidental Spills Application MECHANISM

Herbicide Mixture (before SECONDARY Sediment Surface Water adsorption/dilution occurs) SOURCE

Partioning into Water/ RELEASE Mixing and Dispersion Adsorbing to Sediment MECHANISM

Herbicide Mixture before EXPOSURE Sediment Surface Water adsorption/dilution occurs) MEDIUM a a a

EXPOSURE ROUTE Ingestion Ingestion Ingestion Inhalation Food-Web Exposure Food-Web Exposure Food-Web Exposure Uptake/ Direct Contact Uptake/ Direct Contact Uptake/ Direct Contact

Non-Target Aquatic Plants and C IC IC IC C IC IC C IC IC Algae Aquatic and Benthic IC IC IC IC C C M C C M Invertebrates IC IC IC IC M C M C C M Fish

MCM N M C M M M M Birds

MCM N M M M M M M Mammals Potential Ecological Receptors M M IC M M IC IC M IC IC Humans (Applicators)

Legend: C Potentially complete pathway (evaluated in a quantitative way) N Not quantified because insufficient data available M Minor or insignificant pathway IC Incomplete pathway a Direct contact refers to dermal exposure.

Figure 2. Conceptual Model of Possible Exposure of Biological Organisms to Herbicide Mixture Used by the SpartinaFigure 3.3-2. Control Conceptual Program Model of Possible Exposure of Biological Organisms to Herbicide Mixture Used by the Spartina Control Program pected to be insignificant and unquantifiable due to the nature of the site and frequent movement, ranging habits, and furry or feathery outer skin of most wildlife species. Because Project applications of herbicides would occur only once or (at most) twice a year, and compounds in the herbicide mixture are not expected to persist in significant

San Francisco Estuary Invasive Spartina Project 26. 2010 Aquatic Pesticide Application Plan concentrations for more than several hours, chronic exposure is not likely. In addition, since imazapyr is a newly registered herbicide in California as of 2005, there are few other sources for its introduction to the environment to add to those of the ISP. Therefore, this evaluation focuses on acute toxicity, which would occur when the compounds are present at relatively high concentrations during and immediately following application. Herbicide solutions have the potential to affect organisms that live in the water column, including algae, non-target plants, fish and aquatic invertebrates. While some other receptors such as mammals and birds may spend a considerable portion of their time in the water, they are generally more likely to be affected by other exposure routes, primarily dermal contact during application and incidental ingestion of contaminated sediment during foraging.

Non-Target Aquatic Plants and Algae Due to their engineered mechanisms of action, imazapyr and glyphosate are toxic to a wide variety of plants. Native salt marsh plants, aquatic macrophytes, and algae in the Estuary waters where the herbicides would be applied could be negatively affected. However, both imazapyr and glyphosate are ineffective for treating submerged aquatic vegetation. Imazapyr. The species most sensitive to technical grade imazapyr and an herbicide/surfactant mixture appear to be aquatic macrophytes with reported EC25 values for duckweed (Lemna gibba) of 0.013 mg/L for growth and for common water milfoil (Myriophyllum sibiricum) of 0.013 mg/L for shoot growth and 0.0079 mg/L for root growth. (Hughes 1987; Roshon et al. 1999; both in SERA 2004.) Aquatic algae appear to be substantially less sensitive. The most sensitive species of algae tested was a unicellular green algae (Chlorella emersonii) with an EC50 of about 0.2 mg/L for growth. Some algal species appear to be stimulated rather than inhibited by imazapyr concentrations of up to 100 mg/L. (Hughes 1987 in SERA 2004.) Some species of plants, including aquatic plants, may develop resistance to imazapyr. Bioassays conducted on Chlorella emersonii indicated that resistant strains may be less sensitive by a factor of 10. (Landstein et al. 1993 in SERA 2004.) Due to the infrequent application of imazapyr for control of Spartina, i.e. once per year, development of resistance to imazapyr is unlikely. Recent studies conducted in Washington State also document the potential for imazapyr to impact non-target vegetation. Effects of imazapyr application on non-native Japanese eelgrass were compared to glyphosate application. For both herbicides, the eelgrass canopy was killed if herbicide was applied on dry eelgrass at low tide with imazapyr being more toxic. Application onto an eelgrass bed with a thin overlying film of water did not result in toxic effects. Within 12 months, all treated eelgrass beds had recovered. Persis- tence was not recorded in the sediment underlying these eelgrass beds. (Patten 2003.) Glyphosate. In laboratory growth inhibition studies with submerged aquatic plants no adverse effects on the growth of elodea (Elodea canadensis), water milfoil (Myriophyllum spicatum), and wild celery (Valisneria americana) were found with glyphosate concentrations of up to 1 mg/L. (Forney & David 1981 in WS FEIS 1993.) These results are consistent with the findings of other investigators who report that submerged plants are either resistant or affected only by very high glyphosate concentrations. (Evans 1978; Peverly & Crawford 1975; both in WS FEIS 1993.) A large number of studies with a variety of green algae, blue-green algae, diatoms, and periphyton indicate that glyphosate is

San Francisco Estuary Invasive Spartina Project 27. 2010 Aquatic Pesticide Application Plan slightly toxic to practically non-toxic to most algae. Most algae tolerate concentrations of glyphosate greater than 1 mg/L. (WS FEIS 1993.) Species of algae vary in their sensitivity to glyphosate in terms of population growth (EXTOXNET, Giesy 2000). Field studies indicate the least toxicity to phytoplankton (microscopic floating algae), possibly because of dilution and adsorption in open water and flooded marshes. Few data are available on effects to marine algae, as most toxicity tests have been performed on freshwater species. Giesy et al. (2000) reviewed the data available on glyphosate toxicity to micro-organisms, and found that acute toxicity EC50 values ranged from 2.1 to 189 mg/L. NOECs ranged from 0.73 to 33.6 mg/L. Giesy et al. (2000) also reviewed the data available on glyphosate toxicity to aquatic macrophytes, and found that acute toxicity EC50 values ranged from 3.9 to 15.1 mg/L. It should be noted that these studies included tests on the (non-aquatic) Roundup formulation as well as other forms of glyphosate. The formulated product known as Roundup, which includes the terrestrial surfactant polyethoxylated tallowamine (POEA) is known to be more toxic than the technical grade glyphosate found in aquatic formulations such as Aquamaster. For studies conducted on microorganisms using glyphosate tested as isopropylamine salt, EC50 values ranged from 72.9 to 412 mg/L, and NOEC values ranged from 7.9 to 26.5 mg/L (Giesy et al. 2000). The lowest of these NOEC values (0.73 mg/L) is well above the maximum concentration of 0.026 mg/L reported by Paveglio et al. (1996) and the immediate maximum geometric mean glyphosate concentration of 0.174 mg/L reported by Patten (2002). Therefore, these data indicate that impacts to non-target submerged aquatic plants or algae are not likely. Impacts in estuarine conditions with high concentrations of suspended sediment, which interfere with glyphosate activity, would be even less likely.

Aquatic and Benthic Invertebrates Imazapyr. Imazapyr has been found to have low toxicity to aquatic invertebrates. A study where Daphnia was exposed to an imazapyr formulation (~50%) produced a 48-hour EC50 concentration of 373 mg imazapyr a.e./L (Cyanamid 1997 in Entrix 2003.). Another study with Arsenal® (identical to Habitat®) with an unspecified surfactant determined a 48-hour LC50 of 350 mg Arsenal/L (79.1 mg imazapyr a.e./L) and a NOEC of 180 mg Arsenal/L (40.7 mg imazapyr a.e./L) for the freshwater flea (Daphnia magna), highlight- ing the potential effects of surfactants on aquatic toxicity. Other studies also reported 24 and 48-hour LC50 concentrations of greater than 100 mg/L, the highest dose tested (“HDT”), in static tests conducted with newly-hatched Daphnia. (Kintner & Forbis 1983 in SERA 2004.) Chronic studies reported no adverse effects on survival, reproduction or growth of 1st generation Daphnia after 7, 14 and 21-days of exposure at concentrations up to 97.1 mg/L, the HDT. (Manning 1989 in SERA 2004.). Testing with other invertebrate species that exhibit alternative life cycles has been limited to survival of pink shrimp (Penaeus duorarum) and growth studies with the Eastern oyster (Crassostrea virginica). Acute toxicity to pink shrimp was determined at LC50 >132 mg imazapyr a.e. /L, the HDT, which was also the NOEC. The EC50 for growth inhibition of the Eastern oyster was established at a concentration greater than 132 mg imazapyr a.e./L, with the NOEC set at this concentration, the HDT. (Mangels & Ritter 2000 in SERA 2004.) A recent microcosm study analyzing benthic macroinvertebrates in a logged pond confirmed the low toxicity of imazapyr to benthic freshwater macroinvertebrates. The study

San Francisco Estuary Invasive Spartina Project 28. 2010 Aquatic Pesticide Application Plan analyzed macroinvertebrate community composition, chironomid deformity rate, and chironomid biomass and concluded that imazapyr did not affect the macroinvertebrate community at the concentrations tested. The NOEC was determined to be greater than 18.4 mg/L (Fowlkes et al. 2003.) Glyphosate. Glyphosate is only slightly toxic to practically non-toxic to marine and freshwater aquatic invertebrates. Acute toxicity for freshwater invertebrates varies from 545 to 780 mg/L for water flea (Daphnia magna), to 673 mg/L for mosquito 4th instar (Anopheles quadrimaculatus), to 1,157 mg/L for a leech (Nephaelopsis obscura). Acute toxicity for marine invertebrates was reported as greater than 10 mg/L for Atlantic oyster larvae (Crassostrea virginica), 281 mg/L for grass shrimp (Palaemonetes vulgaris), and 934 mg/L for fiddler crab (Uca pugilator). (ExToxNet 2005; Henry 1992, Heydens 1991; both in SERA 2004.) The wide variation in the aquatic toxicity of glyphosate has been attributed to the dilution water, temperature, formulation, and the amount of suspended sediment in the water. Toxicity appears to increase with temperature, and decrease with elevated pH and suspended sediment. (Schuette 1998). Giesy et al. (2000) reviewed the data available on glyphosate toxicity to aquatic invertebrates. Few data were available for marine species, and those studies that did use marine species were conducted with glyphosate acid, not salt. Acute toxicity EC50 values for five marine species ranged from 281 mg/L to greater than 1000 mg/L, and NOEC values ranged from 10 to 1000 mg/L. Data compiled by Ebasco (1993) include mortality tests on two marine species, for which EC50 values were found to be 281 mg/L and greater than 1,000 mg/L. Grue et al. (2002) conducted laboratory studies to evaluate reproductive effects of exposure to Rodeo mixed with four different surfactants, including R-11, LI 700, and Agri- dex, on Pacific oysters. The EC50 for glyphosate alone was 68.1 mg/L, the EC50 for the tank mix including Rodeo and R-11 surfactant was 29.9 mg/L, and the EC50 for the R-11 surfactant alone was 1.0 mg/L.

The lowest of these NOEC and LC50 values (10 mg/L) for glyphosate or glyphosate/surfactant mixtures is well above the maximum glyphosate concentration of 0.026 mg/L reported by Paveglio et al. (1996) and the immediate maximum geometric mean glyphosate concentration of 0.174 mg/L reported by Patten (2002). Therefore, these data indicate that impacts to aquatic invertebrates due to post-application water concentrations of glyphosate are unlikely in experimental conditions. Impacts in estuarine conditions with high concentrations of suspended sediment, which interfere with glyphosate activity, would be even less likely. Kubena et al. (1997) conducted sediment and water toxicity studies on marine invertebrates (oysters and amphipods). The LC50 values for Rodeo and surfactant in water ranged from 200 to 400 mg/L, and the LC50 values in sediment ranged from 1000 to 6000 mg/kg. These LC50 values are well above the highest measured geometric mean sediment concentrations of 2.3 mg/L reported by Kilbride et al. (2001) and Patten (2002). Field studies of glyphosate/surfactant applications to tidal mudflat invertebrate communi- ties in Willapa Bay, Washington, agree with laboratory tests, which indicate low potential for adverse impacts to benthic invertebrates. Sampling of benthic invertebrates in mudflats up to 199 days after glyphosate/surfactant (X-77) applications revealed no short- term or long-term effects. Short-term laboratory tests of amphipods exposed to gly-

San Francisco Estuary Invasive Spartina Project 29. 2010 Aquatic Pesticide Application Plan phosate and surfactants did not affect survival even at high concentrations relative to post-spray field conditions (Kubena 1996).

Fish Imazapyr. As detailed in both the 2003 Entrix and 2004 SERA reports, a number of standard bioassays submitted to the U.S. EPA in support of the registration of imazapyr indicate very low toxicity to fish with 96-hr LC50 values greater than 100 mg/L in most studies. According to U.S. EPA’s ecotoxicity classification for aquatic organisms (see Table A-6), these values classify imazapyr as practically non-toxic, the lowest category for addressing acute risk to aquatic organisms from use of chemicals. (U.S. EPA 2005.) A recent study suggests that both Habitat® and Rodeo® have relatively low toxicity to juvenile ® rainbow trout. The LC50 determined for Arsenal (a terrestrial formulation identical to Habitat® that did not contain any surfactants) was determined at 22,305 mg imazapyr a.e./L. (King et al. 2004.)

One study reported much lower 96-hr LC50 values of 4.7 mg/L for Nile tilapia (Tilapia nilotica) and 2.7 mg/L for silver barb (Barbus genionotus). (Supamataya et al. 1981 in SERA 2004.) Although the herbicide used was not specified, it is likely that a formulation was used rather than the technical grade active ingredient. Historically imazapyr herbicides contained surfactants and a formulation that removed the surfactant was only developed in 1992. The use of an herbicide containing surfactants might explain the considerably lower LC50 values. The 2004 SERA report used the lowest LC50 value from this study, 2.7 mg/L, for their risk assessment despite some reservations about the study due to the fact that they only had access to its abstract and because the species studied were not native to the U.S. Nevertheless, the 2004 SERA report assumed that, even though the study was not well documented, the response of these apparently sensitive species may well encompass the response of other sensitive species native to the U.S. (SERA 2004, p. 4-22.) This conclusion is supported by a study that examined the comparative sensitivity of eight ESA-listed fish species to standard test organisms exposed to five different pesticides or metals in order to validate the use of surrogate species as a predictive tool in toxicological assessments. Based on their findings, the authors concluded that a safety factor of two would provide a conservative estimate in risk assessments for listed cold-water, warm-water and euryhaline fish species. (Sappington et al. 2000 in Entrix 2003, p. 49.) Glyphosate. Acute toxicity studies with warm and cold water fish indicate that technical glyphosate is slightly to practically non-toxic. (U.S. EPA 1993.) Acute toxicity LC50 values were reported at 86 mg/L in rainbow trout, 120 mg/L in bluegill sunfish, and 168 mg/L in harlequin. (ExToxNet 2005.) Chronic toxicity studies with a terrestrial formulation of glyphosate, Roundup®, found no significant adverse effects on growth, carcino- genicity, feeding, and agonistic behavior in rainbow trout fingerlings. The authors concluded that sublethal levels of the formulation are relatively non-toxic. (Morgan & Kice- niuk 1992 in WS FEIS 1993.) A recent study with the aquatic formulation Rodeo® determined the LC50 for juvenile rainbow trout at 782 mg glyphosate a.e./L. Giesy et al. (2000) reviewed the data available on glyphosate toxicity to fish. Although some data were available for anadromous species, it appears that all tests were conducted using freshwater test methods. Acute toxicity LC50 values for glyphosate tested as isopropylamine salt ranged from 97 to greater than 1,000 mg/L and NOEC values ranged

San Francisco Estuary Invasive Spartina Project 30. 2010 Aquatic Pesticide Application Plan from <97 to 1,000 mg/L. Data compiled by Ebasco (1993) on one-day acute toxicity tests indicate EC50 values ranging from 12.8 mg/L to 240 mg/L.

The lowest of these NOEC and LC50 values (12.8 mg/L) for glyphosate or glyphosate/surfactant mixtures is well above the maximum glyphosate concentration of 0.026 mg/L reported by Paveglio et al. (1996) and the immediate maximum geometric mean glyphosate concentration of 0.174 mg/L reported by Patten (2002). Therefore, these data indicate that impacts to fish due to maximum post-application water concentrations of glyphosate are unlikely in experimental conditions. Impacts in estuarine conditions with high concentrations of suspended sediment, which interfere with glyphosate activity, would be even less likely.

Birds Imazapyr. Only few toxicity studies exist for birds. No adverse effects were noted at imazapyr concentrations of up to 5,000 ppm in the diet. Based on the highest doses tested and the U.S. EPA ecotoxicity categories, these results suggest that imazapyr is moderately or less toxic orally to birds. No data exist for the potential toxicity of imazapyr to shorebirds. (Fletcher 1983a,b,c,d in SERA 2004.) No studies exist on toxicity to raptors or on preening or inhalation exposure potentials. Glyphosate. Effects of glyphosate on birds have been tested on mallard ducks (dabbling ducks which ingest wetland sediment along with seeds, insects, and vegetation) and bobwhite quail. Glyphosate is no more than slightly toxic to birds. Several single-dose acute oral studies indicate that glyphosate is practically non-toxic to upland birds and only slightly toxic to waterfowl. (U.S. EPA 1993.) As with mammals, very high dietary concentrations of glyphosate (a 4,640 mg/kg dietary concentration) resulted in no adverse reactions such as weight loss or mortality (Ebasco 1993). Chronic exposure studies with glyphosate determined a no-observed-effect concentration (NOEC) of 1,000 ppm in the diet. (Heydens 1991 in WS FEIS 1993.) Little or no data are available on toxicity of surfactants to birds.

Mammals Imazapyr. Based on U.S. EPA ecotoxicity criteria, imazapyr is considered practically non-toxic to mammals via oral or dermal administration based on acute and chronic studies conducted with a variety of mammalian species. For example, the reported acute oral LD50 for technical imazapyr in rats is greater than 5,000 mg/kg body weight (b.w.) Rats were observed to rapidly excrete imazapyr in urine and feces with no residues detected in their liver, kidney, muscle, fat, or blood. No observable effect was noted for any formulation of imazapyr administered dermally. Very few inhalatory studies were performed and none tested concentrations high enough to determine acute toxicity. Inhalatory effects at sublethal concentrations (<5 mg/L aerosol) were found with technical grade imazapyr resulting in slight nasal discharge and congested lungs. Technical grade imazapyr and imazapyr isopropylamine salt were both found to be moderately irritating to rabbit eyes with complete recovery within 7 days. Technical grade imazapyr is reported as mildly irritating to rabbit skin. Commercial formulations of imazapyr appear to be less toxic via dermal exposure. (Entrix 2003, p. 42-44.) Chronic and subchronic toxicity studies with imazapyr with dogs, mice, and rats did not suggest any systemic toxic or carcinogenic effects. (SERA 2004.)

San Francisco Estuary Invasive Spartina Project 31. 2010 Aquatic Pesticide Application Plan Glyphosate. Glyphosate has been determined to be practically non-toxic to mammals by ingestion with an acute oral LD50 of 5,600 mg/kg b.w. in rats. The no-observed-effect level (NOEL) for chronic toxicity to rats has been determined at 362 mg/kg b.w./day (8,000 ppm) and LOEL at 940 mg/kg b.w./day (20,000 ppm). (USDA 1981; Monsanto 1983; both in WS FEIS 2003.) The reported acute LD50 values for dermal effects range from >5,000 to 7,940 mg/kg for rabbits. Subchronic oral toxicity studies of glyphosate with rats and dogs indicate that oral does of up to 2,000 ppm do not significantly affect behavior, survival, or body weight. Laboratory studies of the chronic effects of glyphosate show that it is slightly to practically non-irritating to rabbits’ eyes. No significant reproductive, teratogenic, mutagenic, or carcinogenic effects from exposure to concentrations of up to 300 ppm were reported in 20-year laboratory studies with rats, dogs, rabbits, and mice. Little is known about potential interactive effects between applied imazapyr or glyphosate/surfactant solutions and cumulative loads of herbicides, insecticides, detergents, perfume agents, and many other organic contaminants in the San Francisco Estuary. It is reasonable to assume that cumulative, interactive effects occur in organisms of the Estu- ary, but the complexity of multiple interactions in uncontrolled field conditions makes definitive research difficult. In practice, total dosages of imazapyr or glyphosate/surfactant solutions applied in field conditions (amount of solution applied, and concentration, and the number of re- applications to eradicate survivors) depends on many factors which are independent of the physiology of imazapyr, glyphosate or the surfactants themselves. The physiological activity and health of the plant, interference with spray coverage by persistent dead leaves or sediment films, all can affect the percent kill of vegetation, and the ability of regenera- tive buds to survive and re-establish the population. Regeneration requires re-application of herbicide or other eradication methods.

7. DESCRIPTION OF ALTERNATE, NON-CHEMICAL CONTROL METHODS The Spartina Control Program implements a number of non-chemical control methods in situations where environmental impacts can be reduced, sites are appropriate for volunteer efforts and educational opportunities, or where public demand dictates. These methods are especially appropriate for small, newly establishing infestations and/or for some of the final cleanup of sites after control efforts have reduced the infestation down to small remaining patches. Non-chemical control methods are also a great way to engage the community around a site because they foster volunteer stewardship efforts and public outreach. During the development of the Site-Specific Plans for control of invasive Spartina around the Bay, the Spartina Control Program evaluated every possible known control method that could be used. A number of criteria were used in the evaluation including efficacy at controlling the Spartina, human health and safety, damage to the marsh habitat and/or other aspects of the environment, impacts on water quality, feasibility of applying the method in the salt marsh, cost, etc. Several non-chemical methods (hand-pulling and manual excavation, covering/blanketing, and mechanical excavation or dredging) have been incorporated into the Site-Specific Plans for 2008-2010. The remainder of the methods that were evaluated were found to have significant limitations, and are not part of the Spartina Control Program’s current plans. However, some of these methods may be used in con-

San Francisco Estuary Invasive Spartina Project 32. 2010 Aquatic Pesticide Application Plan junction with our selected control methods at a later date. The entire set of possible control methods that were evaluated are discussed below, starting with the methods that were selected and incorporated into the plans.

ISP-selected Non-chemical Control Methods

Hand-pulling and manual excavation Manual removal methods are the simplest technology for removal of cordgrass. Manual removal includes pulling cordgrass plants out of marsh sediments or using hand tools such as spades, mattocks, or similar tools to cut away as much cordgrass as possible within reach. Manual removal methods are effective primarily at removing aboveground plant parts, but are less effective at removing belowground rhizomes (the horizontal underground stem that sends out roots and shoots from buds) that rapidly regenerate shoots. Unless digging removes the entire marsh soil profile containing viable rhizomes and buds, its effect is equivalent to pruning (see Mowing, burning, pruning, and flaming, below). The vigor with which remaining rhizomes resprout and regrow is often proportional with the severity of the disturbance. Frequent re-digging and maintenance is needed to exhaust rhizome reserves of energy and nutrition, and the population of buds capable of resprouting. Manual removal is most effective on isolated seedlings, or very young discrete clones (asexually reproduced colonies of cordgrass) or clumps, where they are infrequent. Man- ual excavation in tidal marshes is extremely labor-intensive. Most cordgrass colonies occur in soft mud, where footing needed for digging is impossible or hazardous, even for workers on platforms, mats, or snowshoe-like boots adapted for walking on mudflats. Dug plants with roots left in contact with moist soil may retain viability and regenerate in place or disperse on high tides to establish new populations. As the invasive Spartina populations are reduced around San Francisco Bay, the ISP has increased its use of manual removal methods as part of an Integrated Pest Management strategy and a way to reduce the need for herbicide where appropriate. Manual removal is particularly useful on Spartina densiflora, the secondary invasive cordgrass species in the Estuary which is a discretely-rooted bunchgrass as opposed to the deeply-rooted rhizoma- tous clones of the main problem species, hybrid Spartina alterniflora. The infestations of S. densiflora occur mainly in Marin County with the exception of a few outlier populations in Contra Costa and San Mateo Counties. Many tons of S. densiflora will be removed manually in 2008. Disposal of manually removed material, especially root/rhizome systems, is problematic. On-site disposal in marshes may cause additional marsh disturbance and may result in spread of invasive cordgrass by regeneration of viable roots. Where manual removal occurs next to levees, salt ponds, or other non-tidal environments, local disposal may be feasible. Disposal of manually removed materials may also be accomplished with specialized low-ground-pressure equipment (amphibious vehicles), but the number of passes needed to transport materials also increases marsh disturbance.

Mechanical excavation and dredging Mechanical removal in marshes uses equipment specially designed for working in semi- terrestrial, semi-aquatic wetland environments. Excavation and dredging are accom-

San Francisco Estuary Invasive Spartina Project 33. 2010 Aquatic Pesticide Application Plan plished using (1) amphibious dredges fitted with excavators, clamshells, or "cutterhead" dredges, or (2) excavators working from mats (large wood pile supports placed flat on geotextile fabric placed over the marsh surface). Some locations allow use of conventional shallow-draft, barge-mounted dredging equipment working within reach of marsh from the margins of navigable channels, particularly at high tide. Where cordgrass colonies lie within the limited reach of track-mounted excavators working from levees, mechanical removal also can be performed without entry of equipment to aquatic or wetland environments. Another mechanical technique is maceration or pulverization of soil and plant remains on site using modified agricultural equipment, "chewing" them into particles too small to be viable or regenerate. Floating maceration equipment has been used in inland waterways to control submerged aquatic vegetation. The Control Program may support research and development of this method for use in the baylands environment, and would utilize this method if it were shown to be effective and reliable with mitigable impacts. Mechanical excavation working to the full depth of the rhizome system (up to 1 foot) in tidal marshes has the potential to be significantly more effective than manual excavation. Similarly, maceration techniques that almost completely destroy both aboveground and belowground living mass of cordgrass have high potential effectiveness. Both techniques also have significant limitations in the San Francisco Estuary, however. Excavators working from levees have an inherent limitation of short reach or access distance, usually a working distance of less than 20 feet for the size equipment that typical levees could bear. Floating barges with clamshell or cutterhead dredges, in contrast, would need to work at high tides within about 70 feet of the leading edge of cordgrass vegetation. Excavators have sufficient reach to dispose of excavated marsh soil and biomass in non-wetland areas, on levees, or in aquatic habitats such as salt ponds, which lack vegetation. Heavy equipment often is used within the Estuary's tidal marshes for purposes other than eradication of cordgrass, including removal of large debris hazards and contaminated materials, and construction or maintenance of ditches or canals. Most of this work is done on mats, to distribute the weight of equipment and protect underlying vegetation. These actions are usually aimed at operations that are highly localized (points or narrow alignments) in the marsh, and usually on the relatively firm marsh plain. Even there, equipment may become mired in soft spots, and removal of mired equipment can damage the marsh. In contrast, to maintenance-type work, removal of invasive cordgrass involves a mosaic pattern for operations, and occurs most often in the low marsh and mudflats, which do not eas- ily support mats and geotextile fabrics. Thus, control methods based on excavators working on mats would be most applicable to localized, large patches of invasive cordgrass on the marsh plain. Some tidal flats invaded by cordgrass occur on sandy deltas with intertidal sand bars (e.g., San Leandro, San Lorenzo Creek) where equipment could be staged, but this situation is unusual. The feasibility of using mechanical excavation or dredging methods at a particular location would be determined based on site-specific conditions.

Covering/blanketing This is another technique that is aimed at exhausting the reserves of energy and nutrition in cordgrass roots and rhizomes and increasing environmental and disease stress. Covering typically involves securing opaque geotextile fabric completely over a patch of cordgrass.

San Francisco Estuary Invasive Spartina Project 34. 2010 Aquatic Pesticide Application Plan This excludes light essential to photosynthesis (transformation of solar energy to food energy), and "bakes" the covered grass in a tent of high temperature and humidity. This technique may be used for discrete colonies (clones) where the geotextile fabric can be fastened to the marsh surface securely with stakes for a sufficiently long period of time. High tides, high winds, and tide-transported debris common in tidal marshes often make this difficult or impossible in some situations. Care must be taken to cover the entire clone to a distance sufficient to cover all rhizomes. If rhizomes spread beyond the reach of the blanketing cover, rhizome connections to exposed, healthy stems can translocate (pipe) foods to the stressed, starving connected portions of the clone under the fabric, and increase overall survival. Staking geotextile tents on soft mudflats is very difficult, and may make this method infeasible in many situations. Wrack (piles or lines of drifted debris and detritus from tidal sources) also is capable of smothering cordgrass and other salt marsh plants. Wrack can be created artificially by placing temporary debris piles on the marsh surface, but cannot be stabilized for long (usually no longer than the highest December-January or June tides or winter storm surges). Their duration at any position in the marsh depends on the frequency and height of tides. The lower in the intertidal zone, the less stable the position of a wrack pile is likely to be. This technique would be used only for small colonies, and would depend on locally available accumulations of organic tidal debris.

Other Non-chemical Methods Evaluated

Mowing, burning, pruning, and flaming Cordgrasses are well adapted to disturbances that "crop" or otherwise remove aboveground biomass. A single event that removes living or dead aboveground cordgrass biomass generally stimulates cordgrass growth, and as soon as a cordgrass stand refoliates, it begins to "recharge" its roots and rhizomes with new food reserves. If vegetation is removed with frequency, roots and rhizomes are prevented from regenerating reserves of energy and nutrition and cordgrass begins to die back as its organs of regeneration and storage become exhausted. If the cordgrass is mown close to the mud surface, it also sev- ers the connections between leaves and roots that transport gases to roots growing in extremely anoxic (oxygen-deprived) waterlogged sediment and further stress the plant. Repeated close mowing may be used to increase physiological stress to a point that cordgrass cannot regenerate; frequent burning would have similar effects. The use of pruning, burning, and mowing for cordgrass eradication in open mudflats and marshes would require very frequent treatment of all aboveground growth until the cordgrass rhizome/root systems become exhausted. For robust stands of Atlantic smooth cordgrass, this may require weekly treatment for more than one growth season. Controlled burning may be used in some situations to remove vegetation prior to other treatments, or to prevent pollen and seed dispersal in founder colonies invading new sites. Burning would be used only in suitable locations, and only during periods of low-wind conditions (especially early morning), when fire hazards in succulent vegetation of tidal pickleweed marshes would be manageable. Ignition, however, may be difficult in cordgrass stands on mudflats.

San Francisco Estuary Invasive Spartina Project 35. 2010 Aquatic Pesticide Application Plan Selective pruning (partial mowing with "weed-whackers" or flaming with hand torches) may be used to remove flowerheads and seedheads of discrete colonies to prevent flow of pollen from contaminating seed production of native cordgrass, and to prevent seed production within founding colonies. However, pruning would have little or no effect on the clone's growth rate and must be followed up with other methods to control spread. Mown vegetation without viable seeds or propagules may be left in place or removed from the site. Vegetation containing viable seeds or propagules would require removal from the treatment site and disposal in a suitable area not conducive to cordgrass growth.

Crushing and mechanical smothering This method uses amphibious track vehicles to trample new plant shoots and stems, and cover them with a layer of sediment. The objective is to smother the plant by preventing the use of stems to transport oxygen to its roots and rhizomes. The method would typically be used in the fall, and ideally a period of time after mowing, when young shoots and stems have developed. This method has been used with some success in Washington State, but has not yet been used in the San Francisco Estuary.

Flooding and draining Flooding and draining techniques entail constructing temporary dikes or other structures to impound standing water or remove water to kill emergent vegetation. Cordgrasses are intolerant of permanently flooded or stable, dry conditions, and are generally absent in the diked nontidal salt marshes of the Estuary. Salt evaporation ponds, managed waterfowl ponds, and completely diked pickleweed marsh exclude cordgrasses, native and non-native alike. Atlantic smooth cordgrass and English cordgrass are capable of invading tidal marsh pools (salt pans) subject to irregular tidal influence (Campbell et al. 1990, P. Baye, pers. observ.), but they are not likely to survive in typical diked wetlands. When tidal marshes are diked and drained rather than flooded, they undergo rapid physical and chemical changes. Organic matter decomposes when microbes are exposed to air; clays shrink when dewatered; and sulfides formed in oxygen-free mud transform to sul- fates forming strong acids (Portnoy, 1999). Therefore, diking and draining, although con- ceivably effective for killing cordgrass, would adversely impact marsh soils and restoration, and the longer salt marsh soils are diked and drained the more difficult these adverse soil changes are to reverse. For these reasons, diking and draining only would be used in critical situations where no other method is feasible, and only after careful evaluation and planned mitigation. Diked salt marsh soils that remain permanently flooded undergo relatively slower and less significant changes. Diked flooded salt marshes would eliminate existing standing vegetation, but are readily re-colonized by youthful salt marsh vegetation if the diking is brief. Isolating the treatment area for flooding or draining may be accomplished by constructing temporary dikes or by closing openings in existing dikes. Temporary constructed dikes need not be large to accomplish treatment. Low earthen berms (about one foot above marsh plain elevation), constructed using low-ground pressure amphibious excavators, could be built around large colonies of cordgrass within open marsh plains. Alternatively, water-filled geotextile tubes ("inflatable dams"), analogous with inflatable cofferdams used in aquatic construction/dewatering operations, may be used. Upon completion of treatment, berms would be graded down to marsh surface elevation, and inflatable dams

San Francisco Estuary Invasive Spartina Project 36. 2010 Aquatic Pesticide Application Plan removed. Temporary dike structures may be difficult to construct in tidal mudflats. Mud- flat sediments are usually too soft to "stack" into berms, and firmer material placed on fluid or plastic muds simply subsides into the flats. Similarly, inflatable dams may not be feasible for softer tidal flats. Many populations of non-native cordgrasses have invaded marshes restored by breaching dikes within former diked baylands, where most of the original dikes remain. In these situations, a dike-enclosed tidal marsh could be temporarily re-closed ("choked") by placing a sheetpile barrier in the existing breach, thus creating a temporary lagoon and effect- ing mass cordgrass eradication. Water control structures (adjustable tidegates) may be installed to enable marsh managers to maintain water depths lethal to cordgrass, suitable diving duck habitat, and adequate water quality. Marsh recolonization is expected to proceed rapidly following restoration of tidal flows. An alternative form of treatment, intermediate between flooding and draining, would be to combine impoundment of water with deliberate solar evaporation, creating hypersaline lagoons. Hypersaline conditions would make the habitat transformation even more rapidly lethal for invasive cordgrass. Restoring tidal flows to temporary salt ponds, however, may require dilution of brines, which could increase cost.

8. HERBICIDE APPLICATION AREAS FOR 2008-2010 Twenty-five sites, comprising 168 sub-areas and less than 75 acres of non-native Spartina, are slated for control during the 2010 Spartina Treatment Season (Figure 3), and approximately 146 of these infestation sites will potentially be treated with herbicide for at least a portion of their control work. Most of these sites are being re-treated for the third to fifth year, hence the significant reduction in treatment area from 2,000 acres in 2006. In the cases where it will be utilized, aquatic herbicide was determined to be the most efficacious and least impactful method for Spartina control following a thorough evaluation of the available control methods described above. Each site has been evaluated taking into account many factors including site access, endangered species issues, habitat sensitivity, the infestation’s susceptibility to non-chemical control methods, partner involvement, public comment, and other factors. A brief description of each site follows including the herbicide delivery systems(s) to be used and an identification of the ISP Partner involved in treatment work on the site. Detailed maps of the each site can be found in Appendix 2.

Site 01 – Alameda Flood Control Channel, Alameda County The Alameda County Flood Control Channel (ACFCC) is a large, unlined, trapezoidal channel that runs from east to west through Hayward, Alameda County, draining a nearly 800 square mile watershed into the San Francisco Bay. The levees on both sides of the ACFCC are topped with multi-use public trails that are part of the San Francisco Bay Trail and Coyote Hills Regional Park. Downstream from Ardenwood Blvd., beyond the levees to the north, are both active and inactive commercial salt ponds, with an industrial facility and parking lot along the furthest upstream levee. To the south are active salt ponds, seasonal wetlands, and Coyote Hills Regional Park. There are currently no hous- ing units, schools or other similar facilities within this lower reach. The combined infestation of the six sub-areas of the Alameda Flood Control Channel comprised one of the largest Spartina alterniflora hybrid infestations in San Francisco

San Francisco Estuary Invasive Spartina Project 37. 2010 Aquatic Pesticide Application Plan Figure 3. Location of 2008-2010 Spartina Treatment Sites within the San Francisco Estuary and adjacent Outer Coast marshes. Each treatment site is comprised of one to 23 sub sites, which are identified by letters (a through z) in project plans and documents

Bay. The ISP's 2004 mapping effort estimated a total of roughly 200 contiguous acres of S. alterniflora/ hybrids on this site spread over approximately 470 acres (43%) of salt marsh and tidal mudflats. The treatments from 2005 -2009 have reduced the infestation by at least 98%. The treatment method at this site is aquatic herbicide, which will be applied by tracked amphibious vehicle and backpack sprayers as the infestation has now been reduced below the level of broadcast aerial. Partners on this site include the Ala- meda County Flood Control District and the California Wildlife Foundation. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

San Francisco Estuary Invasive Spartina Project 38. 2010 Aquatic Pesticide Application Plan Site 02 – Bair & Greco Island Complex, San Mateo County The Bair & Greco Island complex encompassed by this plan is located in the southwest portion of the San Francisco Bay Estuary. The northern edge of the complex is at Bel- mont Slough on the border of Foster City and Redwood City, including the marshes of Brewer Island just south of the San Mateo Bridge. The southern border of the complex is the Union Pacific railroad line just south of the Dumbarton Bridge. The site includes marsh islands, active and inactive commercial salt ponds, six large sloughs with numerous smaller channels, and other bay front marsh that is part of the San Francisco Don Edwards National Wildlife Refuge (DENWR). T he Bair & Greco Island complex contains many different marsh systems, all of which are impacted to varying degrees by Spartina alterniflora hybrids. Of the roughly 3,060 acres of Baylands within the complex, there were approximately 216 acres (7%) infested with non-native Spartina in 2005. Four years of effective herbicide treatments in this complex leave less than 25 acres to treat/retreat in 2010. Three sub-areas were added to this complex for the 2008-2010 Site-Specific Plans, including one along Redwood Creek at Deepwater Slough with a few acres of hybrid Spartina, and Middle Bair which was recently opened to tidal inundation and immediately became infested with scattered hybrid plants. The treatment method at this site is aquatic herbicide, which will be applied by airboat, helicopter, conventional spray truck, backpack sprayer, and tracked amphibious vehicle. Partners on this site include the U.S. Fish and Wildlife Service, Don Ed- wards National Wildlife Refuge and the San Mateo County Mosquito Abatement District. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 03 – Blackie’s Pasture, Marin County Blackie’s Pasture is a small City of Tiburon park located along the shoreline of Richard- son Bay, adjacent to Tiburon Boulevard. With the ample parking provided at the park, it is heavily used by the public for passive recreation. The park is comprised of a 0.7-acre pasture, a small creek channel (“Blackie’s Creek”) along the eastern edge of the pasture, and a shoreline area that includes the channel mouth, a small open mudflat (fed by sediment delivered by the creek), and landscaped pathways and picnic areas. The total area of non-native Spartina at the Blackie’s Pasture site was around one acre in 2005, divided between two sub-areas. This has been reduced to 0.1 acre that will be treated in 2010. The upper portion of this site is represented by the Blackie’s Creek Channel (Sub-Area 3a), wherein Spartina alterniflora hybrids dominated what was previously an open channel. Spartina densiflora and Spartina alterniflora hybrids dominated the mouth of Blackie’s Creek (Sub-Area 3b), readily hybridizing with the native Spartina foliosa stand there. Some excavation of the upper channel occurred in 2005 as part of a restoration project, but only the invasive Spartina in the center of the creek was actually removed; the remainder was treated in 2006. The primary treatment method at this site is manual removal and offsite disposal of the remaining seedlings of S. densiflora, and an application of aquatic herbicide on the hybrid S. alterniflora and that cannot be readily removed by digging. The imazapyr will be applied by backpack sprayer. Partners on this site include the City of Tiburon Public Works and the Tiburon Audubon Society.

San Francisco Estuary Invasive Spartina Project 39. 2010 Aquatic Pesticide Application Plan Site 04 – Corte Madera Creek Complex, Marin County The Corte Madera Creek complex is located on the west side of the North San Francisco Bay in Marin County, south of the San Quentin peninsula and San Rafael Bay. The complex consists of a wide corridor stretching upstream from the mouth of Corte Madera Creek to the uppermost point of non-native Spartina growth in this watershed at about 2.2 miles from the mouth. The entire Corte Madera Creek corridor is heavily developed with upscale residential and commercial facilities. The Corte Madera Creek complex is comprised of 11 distinct sub-areas that occur on both public and private lands, each of which will require different treatment and public outreach approaches. The infestation at the Corte Madera Creek complex was still in a relatively early stage of establishment, with approximately 12 acres scattered over 318 acres, or 4% of the total marsh area. The infestation at this complex contains some unique features, including the only infestation of Spartina anglica in the Bay, as well as the first documented case of Spartina densiflora hybridizing with the native Spartina foliosa (now a relatively common occurrence where both species exist in close proximity). Most of the sub-areas have tiny S. densiflora infestations remaining that allow for purely manual control to be used to complete the eradication. Several sub-areas also have Spartina alterniflora hybrids, bringing the overall site total to three of the four invasive cordgrass species currently found in the Bay. The Conservation Corps North Bay (formerly Marin Conservation Corps) conducted a great deal of manual removal throughout the winters of 2005-2009, and this method was highly effective. All of the sites that shifted from chemical to manual control in 2009 will be revisited this season to manually pull/dig the few plants that have resprouted or emerged from the seedbank, while any healthy Spartina densiflora plants will be treated with imazapyr and glyphosate at the large infestation at Creekside Park and along the Corte Madera Creek mouth behind the Larkspur Ferry Terminal. Partners on this site include the Friends of the Corte Madera Creek Watershed, California Department of Fish and Game, California State Lands Commission, College of Marin, City of Larkspur, Golden Gate Bridge Highway and Transportation District, Marin County Parks and Open Space District, and the Marin County Flood Control and Water Conservation District. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Spe- cific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 05 – Coyote Creek & Mowry Slough Area, Santa Clara County The area encompassed by this Site-Specific Plan includes approximately 3,652 acres of marshland within the San Francisco Don Edwards National Wildlife Refuge that lie between Coyote Creek to the south and the Dumbarton Bridge to the north. The site is surrounded entirely by marsh and salt ponds, and there is very little public access, except for a portion of the Bay Trail along part of Newark Slough (Sub-Area 5c). Within this area, the sub-areas include recently restored tidal marshes, freshwater ponds and upland islands, highly complex and diverse historic marsh habitats that include channels, high marsh, mudflats and pans, thin strip marshes, open mudflats, creek channels and mouths, and sandy beach areas. The pioneering infestation of Spartina alterniflora hybrids in the Coyote Creek and Mowry Slough complex is still very limited in its distribution. However, though the acreage is still relatively small, the clones are distributed throughout the habitat types described above. In

San Francisco Estuary Invasive Spartina Project 40. 2010 Aquatic Pesticide Application Plan sum, these infestations covered approximately 15.3 acres in 2005 scattered over this very large marshland complex, which is equal to just 0.4% of the area. Much of this area was treated for the first time in 2006 using a directed herbicide application tool (spray ball) that the ISP developed with PJ Helicopters of Red Bluff, CA. The results from 2007 treatment were less efficacious and several of these areas need to be treated by broadcast aerial application in 2008. In 2009, a more aggressive treatment strategy was initiated throughout the Refuge in response to increasing discoveries of cryptic hybrids in the extremely valuable habitat. Imazapyr was be applied by airboat, truck, and backpack sprayer, and these methods will be utilized again in 2010 along with the addition of amphibious tracked vehicles (both Argo and MarshMaster) taken from point to point in the marsh where hose will be hauled to the infestation points. The partner on this site is the U.S. Fish and Wildlife Ser- vice, Don Edwards National Wildlife Refuge. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 06 – Emeryville Crescent, Alameda County The Emeryville Crescent marsh is a 103.5-acre, fringing mixed pickleweed (Salicornia virginica) marsh shoreline between Powell Street in Emeryville and the eastern landfall of the Oakland Bay Bridge. Two sub-areas, Emeryville Crescent East (6a) and Emery- ville Crescent West (6b), have been delineated due to the historical ownership and maintenance of the site. The site abuts an extremely heavily developed area on the east side of the Bay, with Interstate 80/580 directly adjacent to the east, and the approach to the San Francisco Bay Bridge adjacent to the south. Local anglers, dog-walkers, and other recreational groups frequently use the marshlands included in this site. Illegal activities such as dumping and littering, unauthorized camping, and public inebriation also occur along the edges of, and sometimes within, the marshlands of this site. The non-native Spartina infestations at Emeryville Crescent were in the early stages of establishment on the site. The individual locations of these plants were scattered across both of the sub-areas, but their overall acreage was small in 2005, at less than 2.6 acres or 2.5% of the combined marsh acreage. The infestation is located mainly along the bay edge of the marsh, adjacent to the open mudflats on the outer edge of the site, with a few scattered clones establishing on the interior portions of the marsh within Sub-Area 6b. Several new hybrid areas appeared since 2008. Treatment in 2010 will be needed on less than 0.1 acre after four years of successful applications. The primary treatment method at this site is aquatic herbicide, which will be applied by conventional spray truck, backpack sprayer, and tracked amphibious vehicle. Partners on this site include the California De- partment of Parks and Recreation and the East Bay Regional Parks District.

Site 07 – Oro Loma Marsh, Alameda County Oro Loma Marsh is a large, 324-acre, recently restored salt pond located on the eastern shore of the San Francisco Bay Estuary adjacent to the town of San Lorenzo, about 1.5 miles south of the Metropolitan Oakland International Airport. The marsh is surrounded by levees, with Bockmann Channel and Sulfur Creek bordering the marsh to the north and south respectively. The San Francisco Bay Trail, a multi-use public recreational pathway, utilizes the levee to the west of Oro Loma, and the Southern Pacific Railroad borders the marsh to the east. The surrounding area includes various industrial and com-

San Francisco Estuary Invasive Spartina Project 41. 2010 Aquatic Pesticide Application Plan mercial developments to the north and south including a sewage treatment plant, electrical substation, and capped landfill. Beyond the railroad to the east are residential developments, the Skywest Golf Course, and Hayward Municipal Airport, with I-880 approximately 0.5 mile from the marsh edge. The entirety of the Oro Loma Marsh site was infested with Spartina alterniflora hybrids across both Sub-Areas prior to the very successful helicopter treatment in 2005, totaling approximately 100 acres (31% of the marsh). This was one of the largest infestations of S. alterniflora hybrids in the eastern Central Bay, and was adjacent to several other large infestations including sites along the San Leandro/ Hayward shoreline. Successful helicopter treatments occurred in 2006 & 2007, and scattered clusters of Spartina throughout the site were treated in 2008 & 2009 by helicopter and airboat. The primary treatment method for 2010 at this site is aquatic herbicide, which will be applied by airboat, Argo and backpacks on the remaining stands. The partner on this site is the East Bay Regional Parks District.

Site 08 – Palo Alto Baylands, Santa Clara County The Palo Alto Baylands is a 1,030-acre nature preserve and park complex owned by the City of Palo Alto and located on the bayfront south of the Dumbarton Bridge. The site includes those areas to the north and south of Charleston Slough, and may extend to the northernmost tip of marshland on the southeast portion of the outlet of Charleston Slough – that land within the city limits of Palo Alto. The site is part of a large expanse of intact native Spartina foliosa and pickleweed (Salicornia virginica)/gumplant (Grindelia sp.) marsh habitat. The park has high visitation on the established pathways through the marsh, and serves as an educational center for birdwatchers, naturalists, local schools, and the public. Other recreational users of the preserve include hikers, kayakers, anglers, bikers, joggers and many others. The pioneering infestation of S. alterniflora hybrids on this site covered approximately one acre in 2005, scattered along the bayfront, mostly south of the Baylands Nature In- terpretive Center, but discovery of numerous cryptic hybrid forms over the years indicates the original infestation was significantly larger. Treatment from 2005-2009 has reduced this infestation by approximately 90%. There are several main areas of infestation: the southeast side of the "Yacht Club Bay" near Mayfield Slough, clones near Hooks Point and Hooks Island, and scattered plants along the bayfront from Mayfield Slough to Charleston Slough. The treatment method at this site is aquatic herbicide, which will be applied by truck & hose, backpack sprayer and boat. Partners on this site include the City of Palo Alto and Palo Alto High School.

Site 09 – Tiscornia Marsh (formerly Pickleweed Park), Marin County Pickleweed Park is an 18-acre City of San Rafael Park located on the edge of San Rafael Bay in the northwestern San Francisco Bay Estuary. It is bounded to the north by San Rafael Creek and to the south by East Canal Street. Bordering the park on the east side is the 10-acre Tiscornia Marsh, a thin band of high marsh pickleweed/gumplant habitat, which grades abruptly from a 2 to 3 foot-high escarpment to an extensive mudflat extending bayward. This band of marshland tapers as it extends southward along the park boundary, and becomes very thin as it curves eastward along the riprap of a levee surrounding an area filled for development. There is an east/west wooden service walkway

San Francisco Estuary Invasive Spartina Project 42. 2010 Aquatic Pesticide Application Plan through the marsh that provides access to the Pacific Gas and Electric (PG&E) power line tower adjacent to the site. There is an establishing population of Spartina densiflora within the fringing marshlands of the park. These plants are at their greatest density in the marsh near the outlet of San Rafael Creek. The plants form a dense stand on the Bay edge surrounding the escarpment, near an electrical tower. Scattered plants extend southward from this main area along the Bay edge, and then east and southeast to Shoreline Park. The total infestation size of S. densiflora on the site was approximately 2200 ft2. In spring 2005, a pioneering infestation of S. alterniflora/ hybrid was also discovered just south of the PG&E walkway. Treatment from 2005-2009 has been highly effective and just a handful of plants will need to be treated in 2010. The primary treatment method at this site is manual removal of all S. densiflora, with very limited aquatic herbicide application by backpack sprayer to several patches of hybrid S. alterniflora. Partners on this site include the Marin Audubon and City of San Rafael.

Site 10 – Point Pinole Regional Shoreline, Contra Costa County Point Pinole Regional Shoreline is a 2,315-acre park owned by the East Bay Regional Parks District. It is located to the west of the city of Richmond, in Contra Costa County. Point Pinole opened to the public in 1973 after the property was acquired from Bethle- hem Steel. Bethlehem had acquired the land in the early 1960s from Atlas Powder Co., one of several firms that had manufactured gunpowder and dynamite there for almost 100 years. The park occupies a short peninsula composed of a main upland core with open, grassy parklands interspersed with predominantly eucalyptus woodlands. Along the northern portion of the park, a wide border of high marsh pickleweed characterizes Whittell Marsh, Sub-Area 10a. The southern portion of the park, Sub-Area 10b is a narrow band of tidal marsh grading quickly over a 10-20 meter span from high marsh pickleweed to sandy mudflat. A new Sub-Area 10c was added in 2008 with EBRPD’s acquisition of Giant Marsh. The non-native Spartina infestations at Point Pinole were in the early stages of establishment. The individual locations of these plants are scattered throughout the sub-areas, but their overall acreage is small. Giant Marsh was first treated in 2008. The primary treatment method at this site is aquatic herbicide, which will be applied by amphibious tracked vehicle. All S. densiflora (primarily in Whittell Marsh) will be removed manually and disposed of off-site. The partner on this site is the East Bay Regional Parks District.

Site 11 – Southampton Marsh, Solano County Southampton Marsh is the largest extant marsh within the Carquinez Strait. Its roughly 175 acres are located within the 720-acre Benicia State Recreation Area, Solano County. Highway 780 borders the park on the north and east, Southampton Bay on the south, and on the west stands the residential development of Vallejo. Cyclists, runners, walkers and roller skaters use the park’s 2.5 miles of road and bike paths, which circle the perimeter of the park. The Marsh lies in the central portion of the park, and consists mostly of high marsh pickleweed/gumplant habitat, with a deep main channel and several smaller channels throughout. Cordylanthus mollis ssp. mollis (soft bird’s-beak), an endangered plant species, can be found in some of the high marsh areas of the site. Access to the marsh is restricted to park personnel and researchers to protect the C. m. mollis populations from potential damage caused by trampling.

San Francisco Estuary Invasive Spartina Project 43. 2010 Aquatic Pesticide Application Plan Southampton Marsh contains the only known population of Spartina patens in the San Francisco Estuary. Several large clones are scattered throughout the southern and western portions of the marsh, and multiple smaller clones are peppered throughout the area. A number of S. patens clones are adjacent to the main channel draining the marsh. There was a total of roughly 0.5 acre of S. patens at this site in 2005, which has reduced by herbicide applications in the areas that are not contiguous with Cordylanthus. Some of the areas occupied by S. patens are directly adjacent to, or interspersed with individuals or patches of Cordylanthus. These areas are highly sensitive, and any S. patens eradication effort here will require diligent protection efforts for the C. m. mollis. For 2010, treatment will occur on only a few hundred square feet, mostly composed of S. patens seedlings. There are also several cryptic hybrid S. alterniflora clones that were first identified in 2008, and these will be added to the control work. The primary treatment method at this site is aquatic herbicide, which will be applied by backpack sprayer. The partner on this site is the California Department of Parks and Recreation.

Site 12 – Southeast San Francisco, San Francisco County The Southeast San Francisco complex includes a scattered group of remnant marshlands totaling 56.7 acres within a heavily industrialized landscape on the western shores of the San Francisco Bay Estuary. The complex is bounded by the Islais Creek Channel and Pier 94 on the north, and the San Francisco County and City boundaries to the south. The Southeast San Francisco complex is adjacent to an inactive naval shipyard, shipping container facilities, and 3Com Park stadium (formerly Candlestick Park), as well as the Bay- view residential neighborhood of San Francisco. Within this area there are a number of marshland habitats including intertidal mudflats, native Spartina foliosa stands, riprap shoreline, marshland fill, and pickleweed-dominated tidal marsh plain. The six sub-areas of the Southeast San Francisco complex contained approximately 8.2 acres of Spartina alterniflora hybrids in 2005. This represents 14.5% of the area of these fragmented remnant marshlands. The rapid hybridization of this population, and conversion of these last pieces of remnant marsh in the area, lends a sense of urgency to the control efforts outlined in this plan. The individual patches of non-native Spartina within this area represent localized ‘stepping stones’ in the available marsh habitat of the area to the open waters of the North Bay, and the outer coast. Two sub-areas were added to this complex following the discovery of these small infestations. Treatment occurred at most of these sites from 2006 to 2009, reducing the area needing treatment in 2010. The treatment methods at this site include a combination of manual digging and application of aquatic herbicide by conventional spray truck or backpack. Partners on this site include the California Department of Parks and Recreation, Literacy for Environmental Justice (LEJ), Port of San Francisco, Golden Gate Audubon, City of San Francisco Recreation & Parks, and the U. S. Navy. Note that this is a complex site composed of multiple sub- areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 13 – Whale’s Tail / Old Alameda Creek Complex, Alameda County The Whale’s Tail and Old Alameda Creek Complex is a 564-acre site on the eastern shores of the San Francisco Bay Estuary, south of the San Mateo Bridge and bordered to the east by Union City. This area includes remnant marshland patches that predate altera-

San Francisco Estuary Invasive Spartina Project 44. 2010 Aquatic Pesticide Application Plan tions to the site for salt production, channelized flood control structures, restored salt pond marshland, small sinuous channels, high marsh flats, mudflats, eroding scarp, sand/shell beach, small depositional deltas, and other habitats. The areas included within this complex are entirely restricted from public access and are either managed by Cali- fornia Department of Fish and Game (CDFG) as wildlife habitat (Sub-Areas 13d, 13e, 13f), or by Alameda County Flood Control District for flood control purposes (Sub-Areas 13a, 13b, 13c, 13g). On the northern side of the main channel, formerly diked salt ponds are undergoing restoration activities to convert them to tidally influenced marshlands. To the south of the main channel, Cargill Corporation maintains active salt-producing evaporator ponds. The invasive Spartina at the Whale’s Tail and Old Alameda Creek Complex is one of the oldest infestations of non-native cordgrass in the San Francisco Estuary. The marshlands of this site contained a total of 82 acres of Spartina alterniflora hybrids representing 14.5% of the area. Applications from 2005 -2009 were very effective, reducing the area to be treated in 2010 to just a few scattered acres, mostly in channels and other low elevation areas. The infestations are approaching eradication in a wide variety of marsh habitats including high marsh pickleweed (Sarcocornia virginica)/saltgrass (Distichlis spi- cata), lower marsh Spartina foliosa/mudflat areas, channel banks, edges of salt pans, and bayfront scarps and mudflats. Since the Cargill Mitigation Marsh was a former salt evaporator pond, it was largely unvegetated with native salt marsh species when tidal action was partially restored in 1995. Without any biotic resistance to invasion, the marsh has become infested with large, coalescing clones of invasive Spartina. On the eastern portion of the site, these clones have coalesced into meadows. The Cargill Mitigation Marsh sub-area contained approximately 19.0 acres of S. alterniflora hybrids, representing 38.8% of this restoration site in 2005. Treatment over the past four years has reduced the infestation substantially, with less than 0.5 acre scattered over the site left to treat in 2010. The treatment method at this site is aquatic herbicide, which will be applied by backpack and truck (successful treatment has eliminated the need for helicopter application for the as of 2009). Partners on this site include the Alameda County Flood Control District, California Department of Fish and Game, and the California Wildlife Foundation. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Spe- cific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 15 – South San Francisco Bay Marshes, Santa Clara County The South Bay Marshes are located at the extreme southern tip of the San Francisco Bay, with both San Mateo and Alameda Counties bordering to the northwest and northeast, respectively. The area includes over 100 miles of shoreline, and encompasses some 1,750 acres of marshland. This highly diverse area includes extensive current and former salt ponds, restoration marshes, creek channels and sloughs, bay fill, large intact salt marshes, brackish marsh areas, slough edge marshes, pans, islands, mudflats, sand/shell beaches and other marsh habitats. Included within this area are Guadalupe Slough, Coyote Creek, Alviso Slough, Mountain View Slough and San Francisquito Creek. There is a high degree of complexity in the South Bay Marshes that will be enhanced significantly by the work of the South Bay Salt Ponds Restoration Project, which will convert sizable portions of former salt-making ponds to various types of marsh habitat.

San Francisco Estuary Invasive Spartina Project 45. 2010 Aquatic Pesticide Application Plan The infestation was approximately 8 acres in 2005, but it had expanded rapidly over the past several years despite the effectiveness of imazapyr, largely due to the cryptic hybrids and post-September 1 treatment. The majority of the infestation occurs in the area of Knapp Tract, on Alviso and Guadalupe Sloughs, and along the bayfront adjacent to the DENWR newly acquired ‘salt ponds’ A1 and A2W. Both Pond A1 and A2W marshes are composed of large expanses of intact native Spartina foliosa (California cordgrass) and pickleweed (Sarcocornia virginica), interspersed with gumplant (Grindelia stricta) and tule (Scirpus sp.) Both marshes lie adjacent to compacted earthen levee roads, which are gated, locked, and require DENWR permission to access. The other sites are patchy and scattered throughout the South Bay extending from San Francisquito Creek in the northwest to the Coyote Slough tidelands in the northeast. Until 2008 the marshes of Faber Laumeister were thought to be free from hybrid Spartina, but surveys and genetic testing showed that this would be an area to focus treatment for the first time in 2009. The primary treatment method at this site is aquatic herbicide, which will be applied by conventional spray truck and backpack sprayer, eliminating the need for any broadcast aerial because of successful treatment in 2009. Partners on this site include the Santa Clara Valley Water Dis- trict and the U.S. Fish and Wildlife Service, Don Edwards National Wildlife Refuge.

Site 16 – Cooley Landing, San Mateo County Cooley Landing is a 165-acre salt marsh restoration site located at the northwestern point of the South San Francisco Bay Estuary, south of the Dumbarton Bridge and adjacent to the point where the Hetch-Hetchy Aqueduct makes landfall on the western shore at Menlo Park. The site is a former salt production evaporator pond that is undergoing restoration to tidal marsh. Initial restoration activities were completed between September and December of 2000, and included the excavation of two breaches through the east levee at locations of historic tidal channels. Re-vegetation of the former salt pond is expected to occur through natural colonization. Performance criteria for the restoration of Cooley Landing requires 70 percent cover of salt marsh vegetation and less than five percent cover of non-native vegetation by the tenth year following restoration. Cooley Landing is part of the Ravenswood Open Space Preserve. Prior to opening Cooley Landing to tidal action in 2000, just five adult clones of invasive Spartina covering a total of 0.1 acre were present along the levees outboard of the restoration area. However, since S. alterniflora hybrids are known to occur on the adjacent properties north and south of the restoration area, and restored salt ponds lack the biotic resistance in the form of an established native plant community, the infestation spread rapidly and already covered 20 acres of the restoration site or 13 % of this large area by 2005. Treatment from 2006-2009 has reduced the infestation by 85%, but a significant amount of hybrid Spartina will be need to be treated in 2010 and this invader is still present to varying degrees throughout the site. The treatment method at this site is aquatic herbicide, which will be applied by airboat, backpack and truck. Partners on this site include S.S. Papadopoulos Associates and the Midpeninsula Regional Open Space District.

Site 17 – Alameda Island / San Leandro Bay Complex, Alameda County The area encompassed by this Site-Specific Plan includes all marshlands of the Alameda and San Leandro Bay Area extending from the western tip of Bayfarm Island and San Leandro Channel in the west, to east of Interstate 880 and the Oakland Coliseum in the

San Francisco Estuary Invasive Spartina Project 46. 2010 Aquatic Pesticide Application Plan east. The northern boundary of the site is the Port of Oakland shipping terminals, and the southern edge is 98th Ave on San Leandro Creek. This area supports many diverse habitat types despite the fact that it is directly adjacent to some of the most highly developed land on the west coast. Within this area there are recently restored tidal marshes, freshwater ponds and upland islands, highly complex and diverse historic marsh habitats that include channels, high marsh, mudflats and pans, thin strip marshes along rip-rapped shoreline, public parks and trails, open mudflats, creek channels and mouths, sandy beach areas, marinas, private residences, commercial areas, industrial manufacturing facilities, shipping, and many other land use types. The Spartina infestations within this site are distributed throughout the habitat types described above. Most notably, Arrowhead Marsh, Fan Marsh and the Elsie Roemer Bird Sanctuary support the largest infestations of Spartina in the Alameda and San Leandro Bay Complex. In sum, the shoreline of this site contained 88.5 acres of non-native Spartina in 2005 targeted for control. This infestation was rapidly expanding into new areas, including the restored marshlands of the Martin Luther King Jr. Wetlands Project. Treatment from 2005 -2009 has reduced this infestation to approximately 15 acres. The treatment method for 2010 at this site complex is aquatic herbicide, which will be applied by conventional spray truck, backpack sprayer, amphibious tracked vehicle, airboat, and helicopter. EBRPD will conduct some “chemical mowing” on the eastern half of Arrow- head, using a low, sub-lethal rate of imazapyr applied by helicopter to stop the plants from setting seed but preserve them as refugia for the large California clapper rail population at this site. Partners on this site include the Alameda County Flood Control District, East Bay Regional Parks District, City of Alameda, City of Oakland, Port of Oakland, U.S. Coast Guard, Save the Bay, and Golden Gate Audubon. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 18 – Colma Creek / San Bruno Marsh Complex, San Mateo County The area encompassed within this Site-Specific plan includes all of the marshlands north of the San Francisco International Airport, up to the northern tip of the outlet of San Bruno Canal or Colma Creek. Within this area there are broad marshlands fringing the industrial fill of South San Francisco, channel bank marshland habitat, open mudflats, pickleweed and gumplant marshes, upland marsh edges, brackish creek channels and other tidal marsh systems. Much of this area is highly developed with light industrial, commercial, and business facilities, and a portion of the Bay Trail runs through the northern portion of the site. For the purposes of this plan, there were an estimated 100.3 acres of marshland within this site in 2005. The Spartina infestations within this site are distributed throughout the habitat types described above. In sum, the marshes of this site contained 60 acres of non-native Spartina targeted for control, representing 60% of the appropriate habitat. This infestation had rapidly expanded onto the open mudflats on the western portion of this site, as well as con- stricting the channels on the northern and southern portions. Non-native Spartina has dominated most of the available marshland habitat, with only scattered populations of native tidal marsh plant species remaining in the area. Because of the unusually large and high density population of clapper rail in this site complex, treatment has been phased over several years, with only the upper channels receiving applications in 2006. The en-

San Francisco Estuary Invasive Spartina Project 47. 2010 Aquatic Pesticide Application Plan tire site was treated in 2007, with approximately 60% just “chemically mowed” with a low, sub-lethal concentration of imazapyr to stop seed production while maintaining the above-ground biomass for rail refugia. In 2008, all remaining hybrid Spartina was treated with the full concentration of imazapyr. This was remarkably successful and less than 5 acres remain scattered over the complex as of 2010, with the western portion of San Bruno Marsh now largely devoid of Spartina after just three seasons. The treatment method for 2010 at this site complex is aquatic herbicide, which will be applied by conventional spray truck, backpack sprayer, amphibious tracked vehicle. For the first time, no broadcast helicopter application is required at this site after the successful treatment strategy that has been implemented over the past several seasons. Partners on this site include the San Mateo County Mosquito Abatement District, San Mateo Flood Control District, City of South San Francisco, and the San Mateo County Transit District (Sam Trans). Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 19 – West San Francisco Bay, San Mateo County The area encompassed by this Site Specific Plan includes all marshlands of San Mateo County extending south from the San Francisco/San Mateo County line in the north to the San Mateo-Hayward Bridge in the South. A separate Site-Specific Plan for the Colma Creek/San Bruno Marsh Complex (TSN:ISP-2005-18) has been developed to specifically address the Spartina treatment approaches for that area, and is therefore not included in this plan. Many of the Sub-Areas for this site are small marshes or mudflat areas bordered by light or heavy industrial development, riprap shoreline, Highway 101, the San Fran- cisco International Airport, or other intensive uses. Several are partially restored marshes. Few of these Sub-Areas support diverse, intact native marsh conditions, and non-native Spartina has come to dominate each one. The infestations of non-native Spartina that constitute the West San Francisco Bay Com- plex are scattered along the shoreline in many types of habitats. Spartina can be found along the rip-rap of shoreline development, in remnant or newly formed pickleweed marsh, along channels emptying into the bay, amongst sand/shell beaches, within large established marsh, in wide lagoons, on shallow mudflats, and in small coves and sheltered crannies all along the Bay edge. In all sub-areas, non-native Spartina was rapidly expanding into the existing available habitat. Out of an estimated 350 acres of marsh habitat covered by this Site-Specific Plan, there were 85 net acres (24%) of non-native Spartina requiring control in 2005. Five years of successful treatment by SMCMAD has reduced this site complex to less than 6 widely scattered acres to be treated in 2010. The primary treatment method at this site is aquatic herbicide, which will be applied by conventional spray truck, backpack sprayer, amphibious tracked vehicle, boat, and helicopter. Site 19 included two sites (Sanchez and Burlingame Lagoons) that contained small infestations of Spartina densiflora that had been transplanted anonymously at some point in the past. Successful imazapyr treatment in 2008 had reduced this S. densiflora infestation to the point of purely manual removal in 2009, and this method will be used again in 2010 to continue the eradication. Partners on this site include the San Mateo County Mosquito Abatement District, San Francisco International Airport, Oyster Point Marina, City of Brisbane, City of Burlingame, City of San Mateo, County of San Mateo, City of Foster City, California State Lands Commission, City of South San Francisco, U.S. Coast

San Francisco Estuary Invasive Spartina Project 48. 2010 Aquatic Pesticide Application Plan Guard Reservation, and a number of individual commercial property owners. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 20 – San Leandro / Hayward Shoreline Complex, Alameda County The area encompassed by this Site-Specific Plan includes the marshlands of the San Leandro and Hayward shoreline, Alameda County, extending south from the Metropoli- tan Golf Course and Oakland International Airport in the north to the San Mateo- Hayward Bridge in the south. A separate Site-Specific Plan for Oro Loma Marsh (TSN:ISP-2004-07) has been developed to specifically address the Spartina treatment approaches for that area, and is therefore not included in this Plan. The marshland areas in this site complex range from large, complex restored marsh systems to channel-bank fringe marsh areas. They line the east shore of the Bay, providing a natural border between the highly urbanized and developed areas of the cities of San Leandro, San Lorenzo, and Hayward and the open waters of the Bay. Much of this area is regularly used for passive recreational activities along portions of the Bay trail, within EBRPD lands, and other trails throughout the area. The infestations of non-native Spartina that constitute the San Leandro and Hayward Shoreline Complex are located along the shoreline in many types of habitats. Invasive Spartina can be found along the rip-rap of shoreline fill and levees, in remnant or newly formed pickleweed marsh, along channels emptying into the bay, amongst sand/shell beaches, within large established marsh restoration sites, on shallow Bay-edge mudflats, and in small coves and sheltered marsh areas along the Bay edge. In all sub-areas, non- native Spartina was rapidly expanding into the existing available habitat. Out of 580 acres of marsh habitat covered by the Site-Specific Plan, there were an estimated 204 net acres of non-native Spartina requiring control in 2005, representing 35.2% of the area. Treatment from 2005 -2009 has reduced this infestation by 90%+, leaving approximately 10 acres to treat in 2010. Eight new sub-areas were added for the 2008-2010 Site-Specific Plans comprised of small newly-discovered infestations within the geographic boundaries of the overall site. The treatment method at this site is aquatic herbicide, which will be applied by airboat, helicopter, conventional spray truck, backpack sprayer, and amphibious tracked vehicle. Partners on this site include the Alameda County Flood Control Dis- trict, East Bay Regional Parks District, City of San Leandro, City of Oakland, and the Oro Loma Sanitary District. Note that this is a complex site composed of multiple sub- areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 21 – Ideal Marsh, Alameda County Ideal Marsh is a 178.8-acre wetland restoration site located on the eastern shore of the San Francisco Bay Estuary that was allowed to naturally restore to unrestricted daily tidal exchange. The site is bordered to the north by the mouth of the Alameda Flood Control Channel (TSN:ISP-2004-1); the shoreline marshes of this site run for approximately 2.5 miles south of the channel to a point about a mile north of the Dumbarton Bridge where a levee cuts back to the shoreline. Levees along the eastern edge of this site separate it from current salt evaporator ponds, with the Coyote Hills, and the Regional Park of the same name, located just over one mile to the east.

San Francisco Estuary Invasive Spartina Project 49. 2010 Aquatic Pesticide Application Plan The two Sub-Areas of Ideal Marsh (North and South) were being rapidly colonized with an expanding infestation of Spartina alterniflora hybrids, with a total of 98 acres of invasive Spartina or 54% of the marshlands at Ideal Marsh in 2006. These plants occupied all habitat types present in these marshes, including open mudflats, sand/shell beaches, eroding marsh edge, pickleweed plain, levee edge and other areas. Helicopter treatment from 2005 -2008 reduced the infestation by 95% or more. The area was treated by backpack for the first time in 2009 with a supporting helicopter application only along the bayfront. The site now contains just tiny patches and individual hybrid S. alterniflora plants scattered across the marsh plain and in a few of the channels, with a moderate infestation still present along the bayfront (possibly related to tidal inundation and dry time). The treatment method at this site is aquatic herbicide, which will be applied by MarshMaster, backpack and truck in 2010. The partner on this site is the U.S. Fish and Wildlife Service, Don Edwards National Wildlife Refuge.

Site 22 – Two Points Complex, Alameda County The Two Points Complex is located on the east side of north San Francisco Bay and southeast San Pablo Bay in Contra Costa County, along the shoreline of the City of Richmond, and includes areas both north and south of the Point Richmond peninsula and the Rich- mond-San Rafael Bridge. The shoreline adjacent to these sites is heavily developed with land use including commercial, industrial, residential, and park or marina areas. The Bay Trail provides recreational access along the upland edge of the southern marshes in the complex, but only provides distant viewing of the north marshes because the trail is located further inland at this point. The marshes of this area are wide fringing marshes with mixed pickleweed/Spartina vegetation assemblages. These marshes contain large and small channels, mudflats, wide mid-marsh plains and other areas. The infestations at the Two Points Complex were still in the early stages of establishment, with approximately 5 acres scattered over the 598 acres of the five sub-areas, or less than 1% of the total marsh area. The establishing clonal patches within this area are located mostly along the bayfront amongst native Spartina stands or along the banks of small channels within the marshes, and are beginning to coalesce. The channels and marsh plain of San Pablo Marsh were being rapidly invaded until treatment began in 2005 on the western portion and 2007 for the eastern. The easternmost portion of San Pablo Marsh was treated for the first time in 2008 because the plants senesced early in both 2006 & 2007. The treatment method at this site for 2010 is aquatic herbicide, applied by airboat, backpack sprayers and Argo. Partners on this site include the State of California Lands Commission, Republic Services, Chevron/Texaco, Richmond Rod and Gun Club Inc., and East Bay Regional Parks District. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 23 – Marin Outliers, Marin County The area encompassed by this Site Specific Plan includes selected marshlands of Marin County extending south from the Point San Pedro in the north to Sausalito in the South. Separate Site-Specific Plans for the Corte Madera Creek Complex (TSN:ISP-2005-04), Blackie’s Pasture (TSN: ISP-2004-03) and Pickleweed Park (TSN:ISP-2004-09) have been developed to specifically address the Spartina treatment approaches for each of

San Francisco Estuary Invasive Spartina Project 50. 2010 Aquatic Pesticide Application Plan those areas. Many of the Sub Areas are small marshes or mudflat areas bordered by light or heavy industrial development, rip-rap shoreline, or other uses. Several are partially restored marshes. In sum, these areas represent an important patchwork of small marsh areas within Marin County. The infestations of non-native Spartina that constitute the Marin Outliers complex are scattered along the shoreline in many types of habitats. Spartina can be found along the rip-rap of shoreline development, in remnant or newly formed pickleweed marsh, along channels emptying into the bay, within large established marsh, on shallow mudflats, and in small coves and sheltered crannies along the Bay edge. In all Sub-Areas, non-native Spartina was in the initial stages of expansion into the existing habitat. Out of an estimated 130 acres of marsh habitat covered by this Site-Specific Plan, there were 2.6 net acres of non-native Spartina requiring control in 2005, representing 2% of the marsh area. Most of these sites are down to only several hundred square feet or less after effective treatments from 2005 -2009. The primary treatment method at a number of sites in this complex is manual removal of S. densiflora after the reductions of the past several control seasons using imazapyr. Other sites will still depend on aquatic herbicide treatment (because of the hybrid S. alterniflora biology), which will be applied by backpack sprayer & conventional spray truck. Partners on this site include Marin Audubon, County of Marin, CDFG, Loch Lomond Marina, Strawberry Recreation District, City of San Rafael, City of Mill Valley, City of Novato, California State Lands Commission, McNear Brick and Block, Paradise Cay Yacht Club, City of Sausalito, and a number of smaller residential and commercial landowners. Note that this is a complex site composed of multiple sub-areas. Please refer to the Site Specific Plan for detailed descriptions of the sub-areas and the associated treatment methods.

Site 24 – Petaluma River, Sonoma County This area includes approximately 4,500 acres of marshland and riparian habitat within the Petaluma River Watershed stretching from the City of Petaluma, at the confluence of the Petaluma River and Lynch Creek in the north to San Pablo Bay in the south. This site consists of a complex mosaic of historic tidal marsh habitat, developed shoreline, brackish tidal riparian edge zones, maintained pastureland, restoration sites, light industrial facilities and urban development. The pioneering infestation of Spartina alterniflora hybrids in the Petaluma River complex was discovered in November 2006, and is still very limited in its distribution. The majority of the infestation is located along the banks of the river adjacent to a dredging and barge dock facility just downstream of the Highway 101 crossing south of Petaluma, with scattered infestations located upstream and downstream this central core. In sum, these infestations covered less than 0.1 acre scattered over this very large marshland complex, which is equal to less than 0.01% of the area. The first treatment at this site occurred in 2007 and follow up work in 2008 & 2009 has reduced this pioneering infestation significantly. The treatment method at this site for 2010 is aquatic herbicide, which will be applied by airboat. The primary ISP partner on this site is the Friends of Petaluma River, a group that has existing relationships with the community of the watershed and the landowners along the river.

San Francisco Estuary Invasive Spartina Project 51. 2010 Aquatic Pesticide Application Plan Site 26 – North San Pablo Bay, Solano County There are two main portions of this sub area: the restoration site on the east side of the Napa River called White Slough, in Vallejo, and the marshes along the shorelines of the Northeastern San Pablo Bay including the mouth of Sonoma Creek. White Slough marsh is a roughly 135-acre restored tidal marsh that lies to the east of Highway 37 and west of Sonoma Boulevard in the City of Vallejo. The marsh is a sparsely vegetated tidal marsh in the initial stages of colonization. The majority of the area is open mudflat with tidally inundated low sections. The periphery of the marsh is composed of scattered pickleweed (Sarcocornia pacifica), non-native Spartina, and alkali bulrush (Bolboschoenus maritimus). The other marsh areas included in this sub-area is the massive mudflat and shoreline area of San Pablo Bay NWR west of Mare Island. This large area of restored and historic tidal marsh, developed shoreline, industrial and military facilities is only lightly infested with non-native Spartina, including both S. densiflora and hybrid S. alterniflora. There is also a newly-discovered infestation at the mouth of Sonoma Creek that consists of several large clones that will be treated by airboat in 2010. All S. densiflora on Mare Island will be removed manually and disposed off site. The infestation at White Slough east of Hwy. 37 was in the very early stages of colonization, and is located along the western, northern and eastern edges of the marsh in scattered small patches. The clonal individuals had not coalesced to any real extent, and there is very little marsh vegetation in some portions of this marsh. Aquatic herbicide was applied by backpack sprayer in 2008 and there may not be any hybrid Spartina to treat here in 2010.

9. WATER QUALITY MONITORING PLAN

Objective Conduct water quality monitoring sufficient to achieve compliance with NPDES State- wide General Permit requirements.

Monitoring Site Selection Approximately 146 sites (sub-areas) of invasive Spartina throughout the San Francisco Bay region are slated for treatment with aquatic herbicide during the 2010 control season. According to current NPDES permit requirements, ten percent (10%), or 15 (rounded from 14.6) of the 146 sites, must be monitored for water quality. These sites are to be selected at representative locations, and are to include all of the herbicides being applied. To assist with sampling site selection, the ISP identified four different treatment site types, as follows: I. Tidal Marsh, Microtidal Marsh, Former Diked Bayland, Backbarrier Marsh II. Fringing Tidal Marsh, Mudflats, and Estuarine Beaches III. Major Tidal Slough, Creek or Flood Control Channel IV. Urbanized Rock, Rip-Rap, Docks, Ramps, etc.

San Francisco Estuary Invasive Spartina Project 52. 2010 Aquatic Pesticide Application Plan The ISP has selected representative sites of each of these marsh site types to be sampled for water quality. Type IV infestation sites are usually very small, sparse, and adjacent to large bodies of water with constant flushing that will serve to quickly dilute any herbicide incidentally entering the water column. For these reasons, along with the fact that ISP has reduced its Type IV sites to tiny infestations of widely-scattered individual plants, none of this site type will be monitored for 2010. Site Types I and II are considered to be the sites most likely to develop detectible levels of herbicide in the water column, although the results of ISP’s 2006 WQMP revealed that the highest one-week post-treatment samples actually came from two Type III sites. For 2010, the final sampling plan list includes eight Type I sites, four Type II sites and four Type III sites chosen to represent the range of herbicide delivery systems and marsh dynamics present in our work program. Imazapyr is the primary herbicide with glyphosate applied to just a handful of the 146 sub-areas that received chemical treatment in 2010); consequently just one glyphosate site will be monitored for water quality as a representative. Table 5 provides a summary of the sites, and the sites are described in a subsequent section.

Sampling Design The sampling events are designed to characterize the potential risk involved with imazapyr and glyphosate applications relative to adjacent surface waters. Consistent with permit requirements, the monitoring program will include background/pre-treatment sampling up to 24 hours prior to the application, application event monitoring immediately post-treatment, and one-week post-application event monitoring. During background sample collection, the point will be recorded using GPS and marked with a flagged PVC pipe to aid ISP staff in locating the point for future sampling events. The application event samples will be collected immediately adjacent to the treatment area after sufficient time has elapsed such that treated water will have entered the adjacent area on the incoming tide. Since it is standard protocol for the ISP partners to treat Spartina on a low or receding tide when possible, application event samples will often be taken 2-5 hours post-treatment when the tide has again flooded the site. Finally, the one- week post-treatment monitoring will be conducted when sufficient water is present at the site on the seventh day after the application. To enhance quality assurance, the ISP will be submitting three to four duplicates over the course of the season for the 48 total base samples taken. These will be added to either the treatment event or one-week post- treatment event since the herbicide levels in the pre-treatment samples are usually ND (no detection). It is standard for the lab to include blanks as part of their quality control, but the ISP will send additional trip blanks on a regular basis. These blanks consist of dis- tilled water taken out in the field and filled at the collection site(s) for that day.

Field Sampling Procedures The Invasive Spartina Project will conduct its own water quality monitoring program for 2010, as it has since 2006. Water samples will be collected using a sampling rod and pre- cleaned amber glass 1-liter bottles. To collect the sample, the bottle is attached to the sampling rod with a clamp, extended out over the water at the application site, and lowered to approximately 50% of the water depth. When the bottle is full it is pulled back out of the water and the cap is affixed to the mouth of the bottle. The sample is labeled in permanent ink with the sample ID number, date, time, and initials of the sampler.

San Francisco Estuary Invasive Spartina Project 53. 2010 Aquatic Pesticide Application Plan Table 5. Summary of Water Quality Monitoring Sites for the 2010 Season

Site Treatment Sites Marsh Type Application Number Date

Creekside Park 4g I 6/28/10 Imazapyr – Backpack Upper Corte Madera 4h III 6/28/10 Imazapyr – Backpack Creek Corte Madera Creek 4j III 7/1/10 Imazapyr – Backpack mouth Imazapyr – Airboat, Argo 7/14/10 San Pablo Marsh 22b I Backpack

Stege Marsh 22d I 7/16/10 Imazapyr – Backpack Blackie’s Pasture 3b III 7/26/10 Imazapyr – Backpack Tiscornia/Pickleweed 9 II 7/26/10 Imazapyr – Backpack Elsie Roemer 17a II 7/28/10 Imazapyr – Backpack Giant Marsh 10c II 8/2/10 Imazapyr – Argo Imazapyr – Truck, 8/12/10 Palo Alto Baylands 8 II Backpack

Bunker Marsh 20g I 8/18/10 Imazapyr – Backpack Sonoma Creek 26c III 8/26/10 Imazapyr – Airboat Stevens Creek Tidal 15c I 9/13/10 Imazapyr – Backpack Marsh Faber Laumeister 15b I 9/14/10 Imazapyr – Backpack Imazapyr – Truck & 9/15/10 Fan Marsh 17j I Backpack

Southampton Marsh 11 I 10/21/10 Glyphosate – Backpack

The sample ID number is determined by the following protocol: a four-letter code unique to the site, followed by the site visit number (e.g., 01 for pre-treatment, 02 for treatment, or 03 for one-week post-treatment), followed by the time since the application (e.g., “pre” for the baseline sample, the number of hours since the application for the treatment sample, or “1w” for the one-week post-treatment).

Equipment Calibration Temperature, electrical conductivity, salinity, and dissolved oxygen will be measured in the field with a portable YSI Model 85 (Yellow Springs Instruments Inc., Ohio, USA), while pH will be measured with an Oakton waterproof pHTestr1 (Oakton Instruments, Illinois, USA). To assure accurate and reliable temperature, electrical conductivity, salinity, and dissolved oxygen measurements, the YSI Model 85 meter will be calibrated, operated, and maintained in accordance with the manual specifications found at http://www.ysi.com/media/pdfs/038503-YSI-Model-85-Operations-Manual-RevE.pdf. To assure accurate and reliable pH measurement, the pHTester 1 meter will be calibrated, operated, and maintained in accordance with the manual specifications found at http://www.4oakton.com/Manuals/pHORPIon/WPpHTestr1_2mnl.pdf. San Francisco Estuary Invasive Spartina Project 54. 2010 Aquatic Pesticide Application Plan Field Data Sheets At each sampling location, the sample ID number, the time of the sampling, the sample depth, and the water temperature, pH, dissolved oxygen, conductivity, and salinity measurements, will be entered on a Field Data Collection Form (“FDCF”, Appendix 3). Also recorded on the FDCF will be site information, including the site ID number, the station location (application point, upstream, downstream), station type (reference, treated), wind conditions, tidal cycle, water color, and the type of herbicide and surfactant that might be present. Any other unusual conditions or concerns will be noted, and any fish, birds, or other wildlife present will be recorded. The FDCFs will be dated and numbered consecu- tively for each site on that date. On return to the office, the data will be entered into an electronic spreadsheet for processing, and the FDCFs will be compiled into a data log and kept permanently in the office.

Sample Shipment Following collection, water samples will be stored on ice packs and shipped for priority overnight delivery to the Pacific Agricultural Laboratory in Portland, OR. If samples are not shipped until the following day, they will be stored in a cooler on ice until they can be transferred to a refrigerator, and subsequently transferred back into a cooler for shipping.

Field Variances The ISP usually selects and plans to monitor two or three more sites each season than is necessary for compliance with the NPDES Permit, to allow for failed sampling events or analyses. If a situation should arise that precludes being able to collect a water quality sample at a designated point at a site at a time suitably close to the specified times (within 24 hours prior to herbicide treatment, within five hours post-treatment, and one week post-treatment), the Water Quality Monitoring Manager (WQMM) will determine whether (1) sampling at the site type is needed to complete the sampling events required by the NPDES permit, or (2) sampling at the site type is not needed for permit compliance, and the site/event can be dropped. If the site type is needed, than the WQMM will consider whether surrogate sampling of some sort (e.g., sampling at a point reasonably nearby the initial point, or at a later or earlier time) would provide an acceptable substitute. If so, the variation will be carefully documented and justified on the Field Data Sheet. If the WQMM determines that surrogate sampling would not be suitable, then an alternate, similar site will be selected and sampled prior to, within five hours post, and within one week post treatment, as a complete replacement for the initial site. Data from samples already collected at the initial site will be kept and reported, along with an ex- planation of the reason for substitution. Any significant problems with sampling events that cannot be remedied in such a way, or any other significant water quality issues that should arise, will be reported to the US EPA Region 9 Project Manager, or their designee.

Sample Analysis The samples will be analyzed within the appropriate holding times for imazapyr (extracted within seven days, analyzed within 21 days of extraction) or glyphosate (within 14 days). Results are reported as parts per billion (ppb), equivalent to μg/L. The analytical method used for imazapyr is EPA 8151A (modified capillary gas chromatograph (GC) for chlorinated herbicides using methylation or pentafluorobenzylation derivation),

San Francisco Estuary Invasive Spartina Project 55. 2010 Aquatic Pesticide Application Plan with a method detection limit (MDL) of 0.2ppb; this is the minimum detectable level of the instrument. However, the reporting limit (RL) for imazapyr, the minimum level that can be quantified with any certainty and accuracy, is slightly higher at 0.5ppb. Gly- phosate is analyzed using EPA method 547 (High Performance Liquid Chromatography with post column derivatization using orthophthalaldehyde (OPA) and fluorescence), with a reporting limit of 5ppb, and for its primary metabolite AMPA 10ppb. The lab runs one blank each time it conducts an analysis (minimum of one sample tested per batch, maximum of three). Results will be submitted to the San Francisco Bay Regional Water Quality Control Board and placed on the ISP’s website for public viewing.

Assessment of Field Contamination Field Blanks. To help asses contamination from field equipment, ambient conditions, sample containers, transit, and the laboratory, one field blank will be collected and submitted for analysis each sampling day. Field blank samples will be obtained by pouring deionized water into a sampling container at the sampling point. Temperature Blanks. For each cooler that is shipped or transported to an analytical laboratory, a 40mL VOA vial will be included that is marked “temperature blank.” This blank will be used by the sample custodian to check the temperature of samples upon receipt by the laboratory.

Lab QC & Data Quality Indicators Each season, the contracted analytical laboratory (“lab”) is required to provide a Quality Assurance Plan (*QAP”) that meets USEPA standards prior to initiating analysis. The lab plan must specify the method of analysis to be used, and describe any variations from a standard protocol. The WQMM will review the lab QAP and determine if it is adequate. At a minimum, the following DQIs will be required for the lab:

Criteria Method Indicator Goal Accuracy of measure- Analyze matrix spikes and 1 matrix spike per 10 samples (10%) ment spike duplicates > 65% @ 2.0 ug/L

Agreement between Analyze lab duplicates and/ Relative percent difference < 25% measurements or matrix spike duplicates Completeness Percent of usable data 95% return (completed/submitted) Comparability of re- Standard reporting units All data reported in micrograms per liter (ug/L) sults or parts per billion (ppb) Use of standardized analy- Standard method used if possible, any modifi- sis methods cations identified, described, and supported. Detection Limits Method detection limit MDL

Monitoring Site Descriptions Following are brief descriptions for each of the monitoring sites. Additional site information was included previously in Section 8 of this plan.

San Francisco Estuary Invasive Spartina Project 56. 2010 Aquatic Pesticide Application Plan Corte Madera Creek Complex. Three sites in this complex will be monitored, the island at the mouth of Corte Madera Creek, treatment along the banks of Upper Corte Madera Creek and Creekside Park. Corte Madera Creek mouth. The island at the mouth of Corte Madera Creek is Type III site located behind the Larkspur Ferry Terminal and north across the creek channel from Greenbrae Boardwalk and the Corte Madera Ecological Re- serve. This island contains the largest infestation of hybrid S. alterniflora in the watershed as a result of seeds being strained out of the incoming tide as propagules enter Corte Madera Creek. This site also contains a moderate infestation of S. densiflora that was difficult to treat properly in the past because seasonal clapper rail restrictions did not allow entry until after the plants had set seed. Both species of invasive Spartina will be treated with a mixture of imazapyr & glyphosate in 2010 with the application to flowering S. densiflora (before seed set) to eliminate any further seed bank accumulation in the substrate. Creekside Park. This Type I site is estimated to contain approximately 21 acres of marshland habitat adjacent to the upper portion of Corte Madera Creek just upstream of Bon Air Road. The park was restored in 1976, when a new channel system was excavated, upland areas were graded to intertidal elevations, and central islands were constructed from the channel dredge spoils as upland refugia. As part of this initial restoration effort, both Spartina densiflora and Spartina anglica were planted, as native marsh plants failed to establish within the first year of restoration. These plants were imported from Humboldt Bay and England respectively. Efficacy from imazapyr applications conducted from 2006 to 2008 controlled the infestation and stopped most seed production, but had not succeeded in full mortality on many of the S. densiflora plants. Beginning in winter 2008-2009, dense areas of the infestation were mowed to bare ground to allow the remaining live plants to sprout fresh healthy growth that can be retreated with a mixture of imazapyr & glyphosate in 2010, while seedlings and other scattered plants adjacent to these invasive meadows will be removed manually as they have since 2005. Upper Corte Madera Creek. This Type III site represents the upper reach from the College of Marin downstream to the Bon Air Road bridge, and includes the short section of an unnamed tributary that drains marshes near Lot 13 (College of Marin parking). Much of this stretch contains narrow strips of marsh vegetation below the rip-rap, mostly pickleweed and S. foliosa, and a mudflat component. Much of the upper reach has an open space character, with Creekside Park and the College of Marin Ecology Study Area along the north bank, a small marsh to the south downstream of the confluence with the Lot 13 drainage, and the backyards of the houses on the south bank set back from the creek on the other side of the Marin County Flood Control District’s gravel maintenance road above the rip-rap. Efficacy from 2006-2009 annual imazapyr applications controlled the infestation but has not produced full mortality on some of the plants. Any green, healthy S. densiflora will be retreated with a mixture of imazapyr & glyphosate in 2010, while plants not healthy enough to translocate the herbicide will be removed manually.

San Francisco Estuary Invasive Spartina Project 57. 2010 Aquatic Pesticide Application Plan San Pablo Marsh. This Type I site is a 165-acre marsh at the mouth of San Pablo Creek on the City of Richmond shoreline in southeastern San Pablo Bay. San Pablo Marsh is predominantly pickleweed with S. foliosa on the bayfront and in the channels. The marsh stretches east to an 11-acre pickleweed, S. foliosa and alkali bulrush (Bolboschoenus maritimus) cove bordered by levees on either side, located behind the Richmond Rod and Gun Club rifle range. The western portion of this site has been treated since 2005, but the larger infestation in the eastern marsh and out onto the mudflats has evaded treatment until recently due to early senescence. The mudflat clones, as well as the infestation en- croaching up the channels, were first treated by airboat and backpack in 2008. Efficacy has been very high on the mudflat clones and in the upper channels, but the established heavy infestation along the fringe has taken longer to control. Airboat and backpack sprayers will be used again at this site in 2010. Stege Marsh. Stege Marsh is a 6-acre Type I site located on the cove of the Richmond Inner Harbor, bordered by the Richmond Marina on the west and Hoffman Marsh (Site 22e) and the Point Isabel Regional Shoreline to the southeast, with I-580 running along the upland edge approximately 500 meters from the marsh, through the City of Rich- mond. The site is part of Eastshore State Park, which is jointly managed by California State Parks and East Bay Regional Parks District (EBRPD). Stege Marsh is a remediation site funded by Cherokee Simeon Venture LLC, which involved excavation and removal of sediments contaminated by former chemical and pesticide manufacturing on the site. New habitat features have also be added as part of the overall restoration, including about 3.5 acres of new marsh habitat and a freshwater lagoon. The Watershed Project has been actively involved in the stewardship and continued restoration of Stege Marsh, including planting pockets of native Spartina foliosa on the mid-elevation mudflats of an inner cove to the north of the Bay Trail that bisects the site. The current infestation at Stege is widely scattered and nearing eradication, but there have been a wide variety of hybrid and native Spartina morphologies over the years that have served to slow progress a bit until the identification was made. The site will be treated with just 2-3 backpack loads in 2010. Blackie’s Pasture. Blackie’s Pasture is a small City of Tiburon park co-managed by the City of Tiburon and Tiburon Audubon. The park is located along the shoreline of Richardson Bay, adjacent to Tiburon Boulevard. The park is heavily used by the public for passive recreation, and is comprised of a 0.7-acre pasture, a small creek channel (Blackie’s Creek) along the eastern edge of the pasture, and a shoreline area that includes the channel mouth, open mudflat (fed by sediment delivered by the creek), landscaped pathways and picnic areas, and rip-rap fill to the east along the Tiburon Peninsula. The Blackie’s Creek channel, a Type III site, was heavily infested with hybrid S. alterniflora, as was the marsh and mudflats at the mouth. The mouth area downstream of the foot- bridge was also heavily infested with S. densiflora. After several years of manual and chemical control work, the infestation in the creek contains just scattered stems and small patches of hybrid S. alterniflora, while the marsh at the mouth is still moderately infested (primarily because of difficulty in hybrid vs. native identification). Treatment in 2010 will involve manual removal of seedlings of S. densiflora and retreatment of hybrid S. alterniflora from backpack sprayers. Tiscornia Marsh/Pickleweed Park. Tiscornia Marsh is a 10-acre Type I site owned by Marin Audubon that borders the City of San Rafael’s Pickleweed Park to the east. This narrow marsh is mostly comprised of thick pickleweed with an eroding 4-5 foot-high es-

San Francisco Estuary Invasive Spartina Project 58. 2010 Aquatic Pesticide Application Plan carpment with an extensive mudflat extending bayward at a very shallow gradient that contains large clones and linear strips of native S. foliosa. This marsh tapers to a very thin strip as it extends southward along the park boundary and as it curves eastward along the riprap. There is a straight, narrow channel running north/south through much of the site that contains both native and hybrid Spartina. An east/west wooden service boardwalk over the marsh provides access to the Pacific Gas and Electric (PG&E) power line tower adjacent to the site. Any seedlings of S. densiflora found on the site will be removed manually while imazapyr will be applied to any remaining patches of hybrid S. alterniflora by backpack. Alameda-San Leandro Bay. Two sites in the Alameda-San Leandro Bay complex will be monitored, Elsie Roemer Bird Sanctuary and Fan Marsh. Elsie Roemer Bird Sanctuary. This Type II site is a composed of a stretch of southern Alameda Island shoreline which runs from the jetty off Shoreline Drive in the west to the Bayfarm Island Bridge in the east. Within the estimated 17.3 acres of marsh at this site, there were approximately 7 acres of non-native Spartina alterniflora hybrids at the start of treatment in 2005, including several large clones coalescing on the open mudflats south of the main infestation. Pro- gress was slow at this site, with low efficacy in 2005 & 2006, and only moderate success from the 2007 imazapyr application. But those three seasons of control work served to weaken this established infestation significantly, and the 2008 & 2009 treatment proved very effective. The remaining hybrid Spartina consists of scattered small stands and individual plants, with pickleweed taking over large areas of the previous infestation. The 2010 applications will be conducted purely by backpack sprayers for the first time in the history of this infestation. Fan Marsh. Fan Marsh is a roughly 11-acre marsh located along on the western side of Doolittle Drive at Earhart Road in Alameda. The property is owned by the Port of Oakland and consists of high marsh pickleweed/Spartina interspersed with several channels draining to the Bay to the south of Doolittle Pond. The Spartina infestation within Fan Marsh was fairly uniform over the entirety of the marsh area. Prior to treatment, roughly 7 acres of Spartina were spread over 11 acres of channels and mid-marsh plain at this site. This Type I site was treated for the first time in 2007, with very high efficacy at the mid-marsh elevation but poor results in the channels at low elevation. Treatment in 2008 & 2009 continued to whittle down the channel infestations that have been more persistent (possibly a dry time issue since the tide floods the site very quickly from San Leandro Bay). The remaining infestation at this site will be retreated where necessary in 2010 using hose from a truck and backpacks. Giant Marsh. Giant Marsh is a 30-acre pickleweed marsh in the far southwestern corner of Point Pinole Regional Shoreline on San Pablo Bay. The infestation at Giant Marsh is composed of scattered clones of hybrid Spartina alterniflora along the bayfront edge of the marsh and out onto the mudflats. This infestation is in the very early stages of establishment (less than an acre), and has not yet colonized the interior of the marsh nor established much along the network of manmade channels. This Type II site was treated for the first time in 2008 and will be retreated in 2010 using an amphibious tracked vehicle and hose. Since the infestation at this site is almost exclusively within the fringe of S. fo-

San Francisco Estuary Invasive Spartina Project 59. 2010 Aquatic Pesticide Application Plan liosa, identification of hybrids has been challenging and has extended the length of time needed to gain control of the infestation. Palo Alto Baylands. This Type II site is part of a 1,940 acre nature preserve and park complex, one of the largest tracts of undisturbed marshland remaining in San Francisco Bay, owned by the City of Palo Alto and located on the western bayfront approximately 2.5 miles south of the Dumbarton Bridge. The site is located east of Hwy. 101 at the end of Embarcadero Road, and includes those areas south of San Francisquito Creek and north of Charleston Slough. Within the site, Harriet Mundy Marsh is a peninsula vegetated with pickleweed (Sarcocornia pacifica), S. foliosa, and gumplant (Grindelia stricta) that extends out to Sand Point from the main parking area. There is a restored marsh cove to the southwest of the parking area that was once home to a yacht club before it was allowed to silt in and return to marshland. The water sample will be taken from the most infested area of the site, Hooks Island, which is located just offshore from Mayfield Slough and contains a heavily channelized pickleweed marsh with large areas of S. foliosa. The site will be treated in 2010 by truck and hose for dense areas near the levee, and with backpack for the remainder (including some island access only by boat).

Bunker Marsh. Bunker Marsh is a large restored marsh sandwiched between residential development in the City of San Leandro to the east and the open San Francisco Bay to the west. This Type I site is a medium-sized marsh on the San Leandro shoreline just north of Robert’s Landing and the San Lorenzo Creek Mouth (Sub-Area-20h). This 32 acre marsh is surrounded by levees and raised berms and is exposed to full tidal action through a wide breach in the levee system on the south side of the marsh. Bunker Marsh contains several habitat types, including open mudflat in the lower central portion of the marsh, small channels, and large sections of mixed Spartina/pickleweed marsh plains. This system was very heavily infested with meadows of hybrid Spartina when treatment began in 2005, but by 2010 the treatment can be conducted by backpack sprayers. Sonoma Creek. This Type III site was discovered late in 2008 and has never been treated due to very early senescence. Several large clones of hybrid Spartina were discovered just upstream of the mouth of Sonoma Creek along the northern shoreline of San Pablo Bay. The banks of the creek are lined with continuous stands of native Spartina foliosa that stretch for miles, and these clones were not observed standing out from the crowd until airboat surveys found them late in summer 2008. These clones will be treated in 2010 by airboat to obtain complete coverage of the full vertical component of these dense stands. Stevens Creek Tidal Marsh. The City of Mountain View’s Shoreline Regional Wildlife and Recreation Area includes several tidal sloughs, bayfront tidal marsh habitat and restored tidal marsh areas. One of the main marshes within this area is Stevens Creek Marsh which has an infestation of hybrid Spartina alterniflora that has been treated since 2007. This Type I site is roughly 30 acres of highly vegetated marsh that was restored from a formerly diked salt pond. The marsh is located on the eastern end of the Recrea- tion Area, at the bayward end of the Stevens Creek Trail. The marsh has well-established populations of native tidal marsh plant species including broad meadows of native Spartina foliosa on the northern half and brackish bulrush stands at the southern end that receives more muted inundation. The majority of the large hybrid clones have been

San Francisco Estuary Invasive Spartina Project 60. 2010 Aquatic Pesticide Application Plan eliminated over the past three seasons, and we will return to the site in 2010 with backpack sprayers to continue the eradication efforts and eliminate the remaining infestation. Faber Laumeister Marsh. This beautiful Type I site owned by USFWS includes the marshlands along the shoreline of East Palo Alto from Bay Road at Cooley Landing south to San Francisquito Creek. This roughly 210-acre complex of tidal marshlands is a remnant patch of a much larger historical marshland community, and maintains a high level of species diversity and habitat complexity. The site is divided into two sections by an earthen levee, with Faber on the south end and Laumeister on the north. The area contains wide meadows of mixed marsh vegetation frequently broken up with sinuous small and large channels lined with dense hedges of Grindelia and native Spartina foliosa. Large populations of the endangered California clapper rail inhabit this marsh, as well as the salt marsh harvest mouse. The site is very lightly infested and was treated for the first time in 2009. We will revisit the site with backpack sprayers in 2010. Southampton Marsh. Southampton Marsh is the largest extant marsh within the Car- quinez Strait. Its roughly 175 acres are located within the 720-acre Benicia State Recrea- tion Area, Solano County. The marsh lies in the central portion of the park, and consists mostly of high marsh pickleweed/gumplant habitat, with a deep main channel and several smaller channels throughout. Cordylanthus mollis ssp. mollis (soft bird’s-beak), an endangered plant species, can be found in some of the high marsh areas of the site. Access to the marsh is restricted to park personnel and researchers to protect the C. m. mollis populations from potential damage caused by trampling. Southampton Marsh contains the only known population of Spartina patens in the San Francisco Estuary. Several large clones were scattered throughout the southern and western portions of the marsh, and multiple smaller clones are peppered throughout the area. A number of S. patens clones are adjacent to the main channel draining the marsh. This is a Type I site and will be the only glyphosate site monitored in this WQMP for 2010.

10. APPLICABLE WATER QUALITY BMPS The following mitigations were identified in the Spartina Control Program’s Program- matic Environmental Impact Report/Statement (PEIR/S). These mitigations will be implemented at all herbicide treatment sites and verified by the Field Operations Staff. IMPACT WQ-1: Degradation of Water Quality Due to Herbicide Application MITIGATION WQ-1: Herbicides shall be applied directly to plants and at low or receding tide to minimize the potential application of herbicide directly on the water surface, as well as to ensure proper dry times before tidal inundation. Herbicides shall be applied by a certified applicator and in accordance with application guidelines and the manufacturer label. The Control Program shall obtain coverage under the State NPDES Permit for the Use of Aquatic Herbicides and any necessary local permits. A monitoring program shall be implemented as part of the NPDES permit, and shall include appropriate toxicological studies to determine toxicity levels of the herbicide solutions being used. The Control Pro- gram shall use adaptive management strategies to refine herbicide application methods to increase control effectiveness and reduce impacts.

San Francisco Estuary Invasive Spartina Project 61. 2010 Aquatic Pesticide Application Plan IMPACT WQ-2: Herbicide Spills MITIGATION WQ-2: Herbicides shall be applied by or under the direct supervision of trained, certified or licensed applicators. Storage of herbicides and adjuvants/surfactants on-site shall be allowed only in accordance with an approved spill prevention and con- tainment plan; on-site mixing and filling operations shall be confined to areas appropriately bermed or otherwise protected to minimize spread or dispersion of spilled herbicide or surfactants into surface waters. When containers of herbicide larger than the standard 2.5 gallon are used (such as the 15 gallon containers that are used by the helicopter contractors for aerial application), these containers must remain in the staging area(s) on a levee or other appropriate upland site. These larger containers will not be allowed into the marsh, and a spill response plan must be in place in the event of an accidental discharge, to ensure that herbicide does not reach the marsh or surface waters. IMPACT WQ-3: Fuel or Petroleum Spills MITIGATION WQ-3: Fueling operations or storage of petroleum products shall be maintained off-site, and a spill prevention and management plan shall be developed and implemented to contain and clean up spills. Transport vessels and vehicles, and other equipment (e.g., mowers, pumps, etc.) shall not be serviced or fueled in the field except under emergency conditions; hand-held gas-powered equipment shall be fueled in the field using precautions to minimize or avoid fuel spills within the marsh. Other, specific best management practices shall be specified as appropriate in project-specific Waste Discharge Requirements. In addition to these water quality mitigation measures, each partner agency and its contractors are required to have an acceptable Site Safety and Materials Handling Plan (Ap- pendix6).

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San Francisco Estuary Invasive Spartina Project 64. 2010 Aquatic Pesticide Application Plan Experimental Station, Dredging Operations Technical Support Program, Technical Re- port D-90-13. Department of the Army, Waterways Experimental Station,Vicksburg, MS. Kroll, R.B. 1991. Field investigations of the environmental fate of Rodeo (glyphosate) in two tidal marshes. Technical Report 115. Maryland Department of the Environment, Balti- more, MD. Kubena, K.M., C.E. Grue, and T.H. DeWitt. 1997. Rounding up the facts about Rodeo: development of concentration-response relationships for selected non-target species. In Proceed- ings of the Second International Spartina Conference, March 20-21. Lee, W.G. and T.R. Partridge. 1983. Rates of spread of Spartina anglica and sediment accretion in the New River Estuary, Invercargill, New Zealand. New Zealand Journal of Botany 21: 231-236. Leson & Associates. 2005. Use of Imazapyr Herbicide to Control Invasive Cordgrass (Spartina spp.) in the San Francisco Estuary: Water Quality, Biological Resources, and Human Health and Safety, prepared for the San Francisco Estuary Invasive Spartina Project, May 4, 2005. Levin, D.A, J. Francisco-Ortega, and R.K. Jansen. 1996. Hybridization and the extinction of rare species. Conservation Biology Volume 10, 10-16. McKee, K.L. and W.H. Patrick. 1988. The relationship of smooth cordgrass (Spartina alterniflora) to tidal datums: a review. Estuaries 11: 143-151. Mendelssohn, I.A. and E.D. Seneca. 1980. The influence of soil drainage on the growth of salt marsh cordgrass, Spartina alterniflora, in North Carolina. Ecology 60: 574- 584Newcombe, C.L., J.H. Morris, P.L. Knutson, and C.S. Gorbics. 1979. Bank erosion control with vegetation, San Francisco Bay, California. Miscellaneous Report No. 79-2, U.S. Army Corps of Engineers, Coastal Engineering Research Center, Belvoir, Virginia. Miller P. and P. Westra. 2004. Herbicide Surfactants and Adjuvants, Colorado State University Cooperative Extension, Bulletin no. 0.559, August 23, 2004. Monsanto Company. 2000. Aquamaster®, Complete Directions for Use in Aquatic and other Non- crop Sites, EPA Reg. No. 524-343, 2000. Munz, P.A., and D.D. Keck, 1968. A California Flora and Supplement. University of California Press. NCAP (National Coalition for Alternatives to Pesticides). 2002. Herbicide Factsheet, Gly- phosate(Roundup). Journal of Pesticide Reform, Caroline Cox, editor. 18:3, Updated September 2002. Rhymer, J.M. and D.S. Simberloff. 1996. Extinction by hybridization and introgression. Annual Review of Ecology and Systematics 27: 83-109. Patten K. 2002. Smooth cordgrass (Spartina alterniflora) control with imazapyr, Weed Technol- ogy, vol. 16, pp. 826-832, 2002. Patten K. 2003. Persistence and non-target impact of imazapyr associated with smooth cordgrass control in an estuary, Journal of Aquatic Plant Management, vol. 41, pp. 1-6. Paveglio, F.L., K.M. Kilbride, C.E. Grue, C.A. Simenstad, and K.L. Fresh. 1996. Use of Rodeo® and X-77® spreader to control smooth cordgrass (Spartina alterniflora) in a southwestern Washington estuary. II. Environmental Fate. Environmental Toxicology and Chemis- try 15. Schuette J. 1998. California Environmental Protection Agency, Department of Pesticide Regula- tion, Environmental Fate of Glyphosate, revised November 1998.

San Francisco Estuary Invasive Spartina Project 65. 2010 Aquatic Pesticide Application Plan SERA (Syracuse Environmental Research Associates, Inc.). 1997. Effects of Surfactants on the Toxicity of Glyphosate, with Specific Reference to Rodeo. Prepared for USDA Animal and Plant Health Inspection Services. SERA, Fayetteville, NY. SERA (Syracuse Environmental Research Associates, Inc.). 2004. Imazapyr - Human Health and Ecological Risk Assessment – Final Report, prepared for USDA, Forest Service, Decem- ber 18, 2004. Smart, R.M. and J.W. Barko. 1978. Influence of sediment salinity and nutrients on the physiological ecology of selected salt marsh plants. Estuarine and Coastal Marine Science 7: 487-495. Spicher, D.P. 1984. The ecology of a caespitose cordgrass (Spartina spp.) introduced to San Francisco Bay. M.A. Thesis, San Francisco State University, San Francisco, California. Sprankle, P., W.F. Meggitt and D. Penner. 1975. Rapid inactivation of glyphosate in soil. Weed Sci. 23:224-228. Tu M, C. Hurd, & J.M. Randall. 2001. Weed Control Methods Handbook: Tools and Techniques for Use in Natural Area, April 2001. U.S. Environmental Protection Agency, Technical Overview of Ecological Risk Assessment, Analysis Phase: Ecological Effects Characterization, Ecotoxicity Categories for Terres- trial and Aquatic Organisms; http://www.epa.gov/oppefed1/ecorisk_ders/toera_analysis_eco.htm#Ecotox, accessed April 2, 2005. U.S. Fish and Wildlife Service and State Coastal Conservancy. (2003) Final Programmatic Envi- ronmental Impact Statement/Environmental Impact Report, San Francisco Estuary Inva- sive Spartina Project: Spartina Control Program. Volume 1: Final Programmatic Envi- ronmental Impact Statement/Environmental Impact Report. State Clearinghouse #2001042058. USFWS, Sacramento, California/State Coastal Conservancy, Oakland, California. www.spartina.org/project_documents/eis_final.htm. September 2003. U.S. Fish and Wildlife Service and State Coastal Conservancy. (2003) Final Programmatic Envi- ronmental Impact Statement/Environmental Impact Report, San Francisco Estuary Inva- sive Spartina Project: Spartina Control Program. Volume 2: Appendices. State Clearing- house #2001042058. USFWS, Sacramento, California/State Coastal Conservancy, Oak- land, California. www.spartina.org/project_documents/eis_final.htm. September 2003. San Francisco Estuary Invasive Spartina Project. 2004. 2004 Site-specific Spartina Control Plans, San Francisco Estuary Invasive Spartina Project: Spartina Control Program. Prepared by SFEISP, Berkeley, California. www.spartina.org/project_documents/. June 2004. Valiela, I., J.M. Teal, W.G. Deuser. 1978. The nature of growth forms in the salt marsh grass Spartina alterniflora. American Naturalist Volume 112 (985) 461-470. Wang, Y.S., C.G. Jaw, and Y.L. Chen. 1994. Accumulation of 2,4-D and Glyphosate in Fish and Water Hyacinth. Water, Air and Soil Pollution. 74(3/4):397-403.

Personal Communications John Smith, BASF, Salem, OR Mark Taylor, East Bay Regional Park District, Hayward, CA Katy Zaremba, Invasive Spartina Project, Stinson Beach, CA

San Francisco Estuary Invasive Spartina Project 66. 2010 Aquatic Pesticide Application Plan APPENDIX 1.

Chemical properties, degradation rates, environmental fate, and toxicity of imazapyr, glyphosate, and aquatic surfactants evaluated for the Spartina Control Program Table A-1: Chemical description; degradation rates, products, and pathways; bioaccumulation ratings; and advantages and disadvantages of imazapyr and glyphosate herbicides for estuarine use

Imazapyr Glyphosate Trade Name Habitat® (Bayer Corporation) Rodeo® (Dow Chemical Company) (Company) Aquamaster® (Monsanto Corporation)

Registration No. 81334-34-1 1071-83-6 Formulation Aqueous solution of isopropylamine salt of imazapyr plus Aqueous solution of isopropylamine salt of glyphosate; acidifier; active ingredient: 28.7% isopropylamine salt of technical formulation contains 2,4-nitrosoglyphosate imazapyr; equivalent to 22.6% imazapyr (“NNG”) impurity; active ingredient: 53.8% glyphosate isopropylamine salt; equivalent to 48.0% glyphosate Chemical name IUPAC: (RS)-2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin- IUPAC: N-(phosphonomethyl)glycine 2-yl)nicotinic acid CAS: N-(phosphonomethyl)glycine CAS: 2-[4,5-dihydro-4-methyl-4-(1-methylethyl)-5-oxo- 1H-imidazol-2-yl]-3-pyridinecarboxylic acid Chemical formula

Formula C13H15N3O3 C3H8NO5P Herbicide family Imidazolinone Organophosphorus Mode of action Systemic, broad-spectrum (non-selective); Systemic, broad-spectrum (non-selective); amino acid synthesis inhibitor, specifically, inhibits amino acid synthesis inhibitor; inhibits acetohydroxyacid synthase (“AHAS”) aka acetolactase 5-enolpyruvylshikimate-3-phosphate synthase, needed by synthase (“ALS”), the first enzyme in the synthesis of plants to synthesize chorismate, an intermediate branched-chain aliphatic amino acids (valine, leucine, and metabolic product in the synthesis of aromatic amino isoleucine) and as a result inhibits protein synthesis and acids cell growth Molecular weight 261.28 g/mole imazapyr 169.08 g/mole glyphosate 320.42 g/mole imazapyr isopropylamine salt 228.22 g/mole glyphosate isopropylamine salt Table A-1 contd.: Chemical description; degradation rates, products, and pathways; bioaccumulation ratings; and advantages and disadvantages of imazapyr and glyphosate herbicides for estuarine use

Imazapyr Glyphosate Specific gravity 1.04–1.07 0.5 Minimum 1 hour 6 hours drying time Highest proposed 1.5 lb a.e./acre 10.8 lb/acre application rate

Rate of kill Very slow Relatively slow Volatility Vapor pressure = 1.8×10-11 mm Hg Extremely low vapor pressure, thus, negligible risk of Henry’s Law constant of 7.1×10-17 atm m3/mole movement through volatility No volatilization from dry soil surfaces; low volatilization of imazapyr from water or moist soil surfaces. Solubility Water: 11,272 mg/L Water: ~12,000 mg/L

Soil organic carbon Koc = 8.81 Koc = 24,000 adsorption coefficient Very low Koc indicates low sorption potential. Very high Koc indicates tight sorption to most soils, suspended solids, and sediments in the environment.

Octanol/water Kow = 0.22, 1.3 Kow = 0.0003 partition coefficient Degradation Slow anaerobic microbial degradation. No degradation Primarily degraded by microbes and fungi in soil or pathways under anaerobic conditions. Rapid photolysis in water. water, under both aerobic and anaerobic conditions. Photodegradation in water and soil are not expected to contribute significantly to glyphosate degradation. Degradation Quinolinic acid Aminomethylphosphonic acid (“AMPA”); further products degraded to carbon dioxide and phosphate.

Half-life in soil t½ = 25–141 days Average t½ = 32 days, based on 47 agricultural and forestry studies. In most cases, >90% degraded within six months after application.

Half-life in t½ = <2 to 7 days t½ = >3 to 12 months benthic sediment

Table A-1, page 2/3 Table A-1 contd.: Chemical description; degradation rates, products, and pathways; bioaccumulation ratings; and advantages and disadvantages of imazapyr and glyphosate herbicides for estuarine use

Imazapyr Glyphosate

Half-life in water No detectable degradation due to hydrolysis up to t½ = 7–14 days 30 days, pH 5-7 Average t½ = 1-4 days (photolysis) Bioaccumulation BCF = 3; BCF in fish after 10-14 day exposure period = 0.2 to 0.3 Low potential for bioaccumulation Low potential for bioaccumulation in aquatic animals; poorly absorbed when ingested by terrestrial mammals; any absorbed glyphosate is rapidly eliminated resulting in minimal tissue retention. Advantages for — Rapid photolysis in water — Low leaching potential due to strong sorption to estuarine use — Shorter minimum drying time than glyphosate soil/sediment particles — No adsorption to particles — Formulation can be mixed with salt water — Aerial applications require an order of magnitude lower spray volumes than glyphosate — Application is more cost-effective than application of glyphosate — Does not require use of non-ionic surfactants Disadvantages for — Increased adverse effects to non-target emerged — Efficacy hindered by minimum drying time estuarine use vegetation due to higher efficacy on vascular plants — Inactivated by adsorption to sediment particles — Formulation requires mixing with freshwater, which is not readily available — Aerial applications require large spray volumes, which require frequent refilling of helicopter tanks — Application is expensive — Requires use of non-ionic surfactants

Table A-1, page 3/3 Table A-2: Chemical properties, environmental fate, general toxicity rating, and toxicity of adjuvants

Adjuvant Degradation General Toxicity (Manufacturer) Ingredients1 Chemical Properties Pathways Toxicity Rating (lowest reported) Non-ionic Surfactants (“NIS”)

R-11® (surface activator) 80% octylphenoxy — soluble in lipid and Slowly biodegraded by Mammals: practically 96-hr LC50, rainbow trout 3.8 ppm2 (Wilbur-Ellis Company) polyethoxyethanol, water progressive shortening of non-toxic orally, mild 96-hr LC50, bluegill sunfish 4.2 ppm2 20% butanol and — flammable ethoxylate chain; skin irritation possible 96-hr LC50, juvenile rainbow trout 6 ppm5 compounded silicone — specific gravity = 1.0 intermediate breakdown Fish: Moderately toxic 48-hr LC50, Daphnia spp. 19 ppm3 products of polyethylene Other aquatic biota: LD50 oral, rabbit >5,840 mg/kg2 glycol (anti-freeze) and slightly toxic LD50 dermal, rabbit >5,000 mg/kg2 short-chain ethoxylates

X-77® (spreader activator) Alkylarylpoly (oxy- — soluble in lipid and Slowly biodegraded by Mammals: practically 96-hr LC50, rainbow trout 4.2 ppm2 (Valent Corp.) ethylene) glycols, free fatty water progressive shortening of non-toxic orally 96-hr LC50, bluegill sunfish 4.3 ppm2 acids, isopropyl alcohol — flammable ethoxylate chain; Fish and other aquatic 48-hr LC50, Daphnia spp. 2 ppm2 intermediate breakdown biota: moderately toxic LD50 oral, rabbit >5,000 mg/kg2 products of polyethylene LD50 dermal, rabbit >5,000 mg/kg2 glycol (anti-freeze) and short-chain ethoxylates

Liberate® (penetrating Phosphatidylcholine — emulsifiable Biodegradation presumed Mammals: practically 96-hr LC50, rainbow trout 17.6 ppm1 surfactant, deposition and (lecithin), methyl esters of — specific gravity = 0.976 rapid due to natural non-toxic orally, NOEC, rainbow trout 12.5 ppm1 drift control agent) fatty acids, alcohol lecithin ingredients moderate skin irritation 48-hr LC50, Daphnia magna 9.3 ppm1 (Loveland Industries, Inc.) ethoxylate possible NOEC, Daphnia magna 7.5 ppm1 LD50 oral, rat >5,000 mg/kg1 LD50 dermal, rat >5,000 mg/kg1

LI-700® (wetting and Phosphatidylcholine — emulsifiable Biodegradation presumed Mammals: practically 96-hr LC50, rainbow trout 17 ppm2 penetrating surfactant) (lecithin), methylacetic — not flammable rapid due to natural non-toxic orally, causes 24-hr LC50, rainbow trout 22 ppm2 (Loveland Industries, Inc.) acid, alkyl polyoxyethylene — specific gravity = 1.03 lecithin ingredients skin and eye irritation 96-hr LC50, juv. rainbow trout 700 ppm5 ether Fish and other aquatic 96-hr LC50, bluegill sunfish 210 ppm2 biota: practically non- 48-hr LC50, Daphnia spp. 170 ppm3 toxic LD50 oral, rat >5,000 mg/kg2 LD50 dermal, rat >5,000 mg/kg2 Cygnet Plus 75% d-limonene and — flammable Mammals: causes skin NOEC, Ceriodaphnia dubia 3.0 ppm4 (Cygnet Enterprises) related isomers, — specific gravity = 0.87 and eye irritation; 96-hr LC50 Ceriodaphnia dubia 6.6 ppm4 15% methylated vegetable Fish: slightly toxic NOEC, rainbow trout 30 ppm4 oil, 10% alkyl hydroxypoly Other aquatic biota: 96-hr LC50, rainbow trout 45 ppm4 oxyethylene; manufactured moderately toxic NOEC, fathead minnow 15 ppm4 from natural limonene 96-hr LC50, fathead minnow ppm4 Esterified Seed Oils (“ESOs”) or Mehylated Seed Oils (“MSOs”)

Competitor® Ethyl oleate, sorbitan alkyl — soluble in water Fish: slightly toxic 96-hr LC50, rainbow trout 95 ppm3 Wilbur-Ellis Company) polyethoxylate ester, — combustible Other aquatic biota: 48-hr LC50, Daphnia spp. >100 ppm3 dialkyl polyoxy-ethylene — specific gravity = 0.9 practically non-toxic glycol Table A-2 contd.: Chemical properties, environmental fate, general toxicity rating, and toxicity of adjuvants

Adjuvant Degradation General Toxicity (Manufacturer) Ingredients1 Chemical Properties Pathways Toxicity Rating (lowest reported) Crop Oil Concentrates (“COC”)

Agri-Dex® (wetting and Proprietary; heavy range — dispersible in water as Biodegradation presumed Mammals: practically 96-hr LC50, rainbow trout 271 ppm2 penetrating agent) paraffin-based petroleum micelles rapid non-toxic through oral 24-hr LC50, rainbow trout 386 ppm2 (Helena Chemical oil with polyol fatty acid — moderately flammable ingestion, mild skin and 96-hr LC50, juv. rainbow trout 271 ppm5 Company) esters and eye irritant; Fish and 48-hr LC50, Daphnia spp. >1,000 ppm3 polyethoxylyated other aquatic biota: LD50 oral, rat 5,010 mg/kg2 derivatives practically non-toxic LD50 dermal, rabbit >2,020 mg/kg2 Silicone-based Surfactants

Dyne-Amic® (activator, Organosilicone , Fish and other aquatic 96-hr LC50, rainbow trout 23.2 ppm3 spreader-sticker, wetting methylated vegetable oil biota: slightly toxic 48-hr LC50, Daphnia spp. 60 ppm3 and penetrating agent, buffer) (Helena Chemical Company)

Kinetic® (spreader-sticker, Organosilicone , Fish and other aquatic 96-hr LC50, rainbow trout 13.9 ppm3 wetting agent) polyoxypropylene- biota: slightly toxic 48-hr LC50, Daphnia spp. 60.7 ppm3 (Helena Chemical polyoxyethylene Company) copolymer Colorants

Blazon® Spray Pattern Proprietary; 30% non-ionic — pH = 7.0 Mammals: practically LD50 rat >5,000 mg/kg1 Indicator “Blue” polymeric colorant, — completely soluble in non-toxic orally; mild (Milliken Chemical) 70% water water skin irritant; not — specific gravity = 1.07 mutagenic — mildly acidic

1 Manufacturer specimen labels 2 Referenced in Entrix 10/03. 3 Erik Johansen, Washington State Department of Agriculture, Memorandum Re: Summary of Acute Toxicity Data for Five Spray Adjuvants, February 4, 2004. 4 Pacific Ecorisk, An Evaluation of the Acute Toxicity of “CYGNET PLUS” to Ceriodaphnia dubia (water flea), Oncorhynchus mykiss (rainbow trout), and Pimephales promelas (fathead minnow), December 10, 2004. 5 King et al. 2004.

Table A-2, page 2/2 Table A-3a: Imazapyr herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Estuary

Application Method Spray Volume Formulation Active Ingredient1 Surfactant2 Colorant 0.25% v/v NIS with 70% a.i.; High volume 0.52-0.75% solution 100 gal/acre 1-1.5 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 3 qt/100 gal handheld sprayer 4-6 pints/100 gal SBS according to label 0.25% v/v NIS with 70% a.i.; Low-volume directed 0.75-1.5% solution 20 gal/acre 0.3-0.6 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 3 qt/100 gal sprayer 1.2-2.4 pints/20 gal SBS according to label 0.25% v/v NIS with 70% a.i.; Broadcast sprayer/ 2.5-7.5% solution 10-30 gal/acre 0.5-1.5 lb a.e./acre ~1% v/v MSO, ESO, or VOC; 0.5-1.5 qt/acre Aerial application 6 pints/10-30 gal SBS according to label

Table A-3b: Glyphosate herbicide mixture component concentrations and application rates for treatment of non-native Spartina in San Francisco Estuary

Application Method Spray Volume Formulation Active Ingredient1 Surfactant2* Colorant

High volume 1-2% solution 100 gal/acre 4-8 lb a.e./acre 0.5% v/v NIS with 50% a.i. 3 qt/100 gal handheld sprayer 1-2 gal/100 gal

Low-volume 1-8% solution 25-200 gal/acre 1.35-10.8 lbs a.e./acre 0.5% v/v NIS with 50% a.i. 3 qt/100 gal directed sprayer 1-8 gal/100 gal

Broadcast sprayer/ 7-40 gal/acre/ 4.5-7.5 pints/acre 2.25-3.75 lb a.e./acre 0.5% v/v NIS with 50% a.i. 0.5-1.5 qt/acre Aerial application 7-20 gal/acre

1 The active ingredient in Rodeo® and Aquamaster® is glyphosate isopropylamine salt; values are expressed as glyphosate acid equivalent 2 NIS = non-ionic surfactant, %v/v = percentage based on volume by volume Table A-4: Worst-case concentration of imazapyr herbicide dissolved in leading edge of incoming tide Assumptions

Worst-case occurs on the leading edge of lateral flow from overtopped channel through an herbicide-treated marsh Herbicide was uniformly sprayed across the entire marsh surface (but not in channels) at an application rate r = 15.6 mg a.e./sqft The herbicide applied on a unit area (1 sqft) is therefore mass m = 15.6 mg a.e. The herbicide dissolves completely in the incoming water A percentage, p, of the herbicide sticks to the vegetation canopy, and does not dissolve in the first one foot of flow depth Incoming tidal water overbanks channel and flows laterally across the surface of the marsh to a maximum distance D Water flow across marsh (after it leaves channel) has a uniform depth d = 1ft A percentage, s, of the active herbicide that was deposited onto the sediment surface dissolves into the water column The dissolved herbicide is instantly fully dissolved in the first unit volume that flows through No evaporation No rain or other input of fresh water

Application rate Habitat® label application rate: 4-6 pints per acre Label indicates 2 pounds imazapyr acid equivalents per gallon Habitat® 6 pints/acre 1.5 lb a.e./acre 2 = 0.75 gal/acre = 15.61 mg a.e./ft

Variables (p, D, and s can be varied): 2 r = 15.61 mg a.e./ft Herbicide application rate 2 m = 15.61 mg a.e. Initial mass of herbicide per unit area (per 1 ft ) p = 0% Percentage of applied herbicide that is absorbed into vegetation canopy d = 1 ft Depth of water flow across marsh (1 ft allows unit volume calculations) a D = 100 ft Distance of lateral flow across the marsh surface s = 60% Percentage of herbicide reaching the sediment that resuspends into water column 3 C = ? Concentration of herbicide in water column (mg a.e./ft )

Equationb C = m × (1-p) × D × s = (mass per unit area) × (1-percent absorbed by plant canopy) × (percent dissolved in water column) × (number of units through which water flows)

3 Computed Concentration C = m 1-p D s = 937 mg/ft 15.61 100% 100 60% 33.1 mg/liter

Notes a) Most Spartina infested marshes in the San Francisco Estuary that will become inundated by tidal water in the days following imazapyr application have a multitude of channels throughout the marsh that will transport water directly from the San Francisco Bay before overbanking and causing lateral flow across the marsh. In these marshes there would be a maximum of 100 feet of lateral flow through sprayed marsh before meeting with another flow. b) Calculation does not take into account potential decay during period of time between spraying and water inundation nor any decay that might occur in water column once the herbicide is resuspended from sediment. Table A-5: Ecotoxicity categories for acute toxicity of pesticides to wildlife1

Mammals Birds Acute Oral or Acute Inhalation Acute Oral Acute Inhalation Toxicity Category Dermal LD50 (mg/kg) LC50 (ppm) LD50 (mg/kg) LC50 (ppm) Very highly toxic <10 <50 <10 <50 Highly toxic 10-50 51-500 10-50 50-500 Moderately toxic 51-500 501-1000 51-500 501-1,000 Slightly toxic 501-2,000 1001-5000 501-2,000 1,001-5,000 Practically non-toxic >2,000 >5,000 >2,000 >5,000

Table A-6: Ecotoxicity categories for acute toxicity of pesticides to aquatic organisms1

Fish or Aquatic Invertebrates Toxicity Category Acute Concentration LC50 (mg/L) Very highly toxic <0.1 Highly toxic 0.1-1 Moderately toxic >1-10 Slightly toxic >10-100 Practically non-toxic >100

Table A-7: Ecotoxicity categories for acute toxicity of pesticides to insects1

Concentration Toxicity Category (μg/bee) Highly toxic <2 Moderately toxic 2 - 11 Practically non-toxic >11

1 U.S. EPA, Technical Overview of Ecological Risk Assessment, Analysis Phase: Ecological Effects Characterization, September 28, 2004. Table A-8: Toxicity of imazapyr to mammals

Animal Administration Testing Facility 3 Test Substance Species Route Gender LD50 or ED50 Effect (Reporting Year) ʑ >5,000 mg/kg b.w. NOEL Rat oral ʐ >5,000 mg/kg b.w. NOEL American Cyanamid ʑ >2,000 mg/kg b.w. NOEL Company (1983)1 Rabbit dermal Imazapyr technical ʐ >2,000 mg/kg b.w. NOEL ʑ >1 ppm ND Food and Drug Research Rat inhalatory >1 ppm ʐ ND Laboratories (1983)1 (analytical)

AC 243,997 (93% pure) Rat inhalation ʑ+ʐ >1.3 ppm L Voss et al. (1983)2

ʑ >10,000 ppm diet DA oral ʐ >10,000 ppm diet DA DA, B, A, S, ʑ 4,200 mg/kg b.w. CY, C, DBW intraperitoneal DA, B, A, S, Rat ʐ 3,700 mg/kg b.w. CY, C, DBW ʑ >5,000 mg/kg b.w. DA subcutaneous Imazapyr ʐ >5,000 mg/kg b.w. DA Medical Scientific isopropylamine ʑ >2,000 mg/kg b.w. NOEL dermal Research, Laboratory technical ʐ >2,000 mg/kg b.w. NOEL (1983)1 (49.3% a.i.) ʑ >10,000 mg/kg b.w. DA oral ʐ >10,000 mg/kg b.w. DA DA, B, A, S, ʑ 3,450 mg/kg b.w. CY, C, DBW Mouse intraperitoneal DA, B, A, S, ʐ 3,000 mg/kg b.w. CY, C, DBW ʑ >5,000 mg/kg b.w. DA, B, S subcutaneous ʐ >5,000 mg/kg b.w. DA, B, S Table A-8 contd.: Toxicity of imazapyr to mammals

Animal Administration Testing Facility 3 Test Substance Species Route Gender LD50 or ED50 Effect (Reporting Year) ʑ >5,000 mg/kg b.w. DA American Cyanamid Rat oral ʐ >5,000 mg/kg b.w. DA Company (1983)1 ʑ >5,000 mg/kg b.w. DA American Cyanamid Mouse oral Imazapyr ʐ >5,000 mg/kg b.w. DA Company (1986)1 isopropylamine ʑ >2,148 mg/kg b.w. NOEL American Cyanamid Rabbit dermal (25% a.i.) ʐ >2,148 mg/kg b.w. NOEL Company (1983)1 ʑ >0.2 NOEL Food and Drug Research Rat inhalatory ʐ >0.2 (analytical) NOEL Laboratories (1983)1 Hershman & Moore Arsenal® 4-AS Rat inhalatory ʑ+ʐ >4.62 ppm L (1986)2

Chopper®RTU (NOS) Rat inhalatory ʑ+ʐ >3.34 ppm L Werley (1987)2

1 cited in Entrix 10/03. 2 cited in SERA 12/04, Appendix 1 3 Acronyms: A = ataxia (loss of ability to coordinate muscular movement); B = blepharoptosis (drooping of upper eyelid); b.w. = body weight; C = convulsion; CY = cyanosis (bluish discoloration of skin and mucous membranes resulting from inadequate oxygenation of blood); DA = decreased activity; DBW = decreased body weight; ED50 = dose causing 50% inhibition of a process; L = lethality; LD50 = lethal dose, 50% kill; ND = nasal discharge; NOEL = no-observable-effect level (no toxic signs); NOS = not otherwise specified; S = sedation Table A-9: Toxicity of imazapyr to birds

Test Test Substance Species (Observed Effect) Result* >1890 mg/kg diet LD , 18-weeks dietary 50 ~200 mg/kg b.w. 1890 mg/kg HDT NOEL, 18-weeks dietary ~200 mg/kg b.w. Northern bobwhite quail >5000 mg/kg diet LD , 5-day acute dietary 50 ~674 mg/kg b.w. 5000 mg/kg HDT Arsenal® NOEL, 5-day acute dietary ~674 mg/kg b.w. (identical with >1890 mg/kg diet Habitat®) LD , 18-weeks dietary 50 ~200 mg/kg b.w. 1890 mg/kg diet NOEL, 18-weeks dietary ~200 mg/kg b.w. Mallard duck >5000 mg/kg diet LD , 5-day acute dietary 50 ~674 mg/kg b.w. 5000 mg/kg HDT NOEL, 5-day acute dietary ~674 mg/kg b.w.

* Fletcher 1983a, 1983b, Fletcher et al. 1984a, 1984b, 1984c, 1984d, 1995a, 1995b; all in SERA 12/04, Appendix 3 Table A-10: Toxicity of imazapyr and imazapyr herbicide/surfactant mixtures to fish

Test Substance + Surfactant Animal Species Test Result Reference Arsenal® Herbicide 96-hr LC 113 ppm surfactant (28.7% imazapyr) + Hasten 50 Smith et al. 20021 Arsenal® Herbicide 96-hr LC50 479 ppm surfactant (28.7% imazapyr) + Agri-Dex® Rainbow trout, juvenile Arsenal® Herbicide (Oncorhynchus mykiss) 77,716 ppm of concentrate Grue 20031 96-hr LC50 (28.7% imazapyr) 22,305 mg imazapyr a.e./L King et al. 2004 ® 43,947 ppm of concentrate Arsenal Concentrate 1 96-hr LC50 Grue 2003 (53.1 a.i. imazapyr) 23,336 mg imazapyr a.e./L AC 243,997 with Cohle & McAllister 96-hr LC >1000 mg/L isopropylamine in water 50 1984a2 Arsenal® Herbicide Bluegill sunfish Cohle & McAllister 96-hr LC 180 mg/L (22.6% purity) (Lepomis macrochirus) 50 1984b2 AC 243,997 Kintner & Forbis 96-hr LC >100 mg/L (99.5% purity) 50 1983a2 Rainbow trout (Salmo gairdneri) Channel catfish Peoples 19842 Imazapyr NOS 96-hr LC >100 mg/L (Ictaluras punctatis) 50 Gagne et al. 19942 Bluegill sunfish (Lepomis macrochirus) Arsenal® Herbicide Cohle & McAllister 96-hr LC 110 mg/L (22.6% purity) 50 1984c2 Rainbow trout Arsenal® Herbicide (Salmo gairdneri) 96-hr LC >110 mg a.e./L Drotter et al. 19952 (21.5% purity) 50 Table A-10 contd.: Toxicity of imazapyr and imazapyr herbicide/surfactant mixtures to fish

Test Substance + Surfactant Animal Species Test Result Reference NOEC 120 mg a.i./L AC 342,997 LOEC >120 mg/L Drotter et al. 19982 (purity NOS) MATC >120 mg/L Fathead minnow 28-day (Pimephales promelas) >118 mg a.i./L AC 342,997 NOEC >118 mg a.i./L Drotter et al. 19992 (99.6% purity) LOEC >118 mg a.i./L MATC Atlantic silverside AC 243,997 (marine) 96-hr LC 184 mg/L Manning 1989a2 (99.5% purity) 50 (Menidia menidia)

24-hr LC50 4,670 μg/L Nile tilapia 48-hr LC50 4,630 μg/L (Tilapia nilotica) 72-hr LC50 4,610 μg/L Supamataya et al. Imazapyr NOS 2 96-hr LC50 4,360 μg/L 1981 Silver barb 24-hr LC50 2,706 μg/L (Barbus genionotus) 96-hr LC50 2,706 μg/L

1 cited in Entrix 10/03 2 cited in SERA 12/04

Abbreviations: LC50 = lethal concentration, 50% kill; LOEC = lowest-observable-effect concentration; MATC = maximum allowable toxicant concentration; NOEC = no-observable-effect concentration (no toxic signs); NOS = not otherwise specified

Table A-10, page 2/2 Table A-11: Toxicity of imazapyr and imazapyr/surfactant mixtures to aquatic invertebrates

Test Substance Species Test Result Reference (observed effect) Arsenal® Applicator’s Freshwater benthic In-situ microcosm Concentrate >18.4 mg/L (HDT) Fowlkes et al. 2003 macroinvertebrates NOEC, (D, BM) (479 g imazapyr a.e./L) Arsenal®Herbicide NOEC 180 mg/L Forbis et al. 19842 (22.6% purity) 48-hr LC50 350 mg/L Arsenal® Freshwater water flea 48-hr LC 79.1 mg imazapyr a.e./L + unidentified surfactant (Daphnia magna) 50 Cyanamid 19971 NOEC 40.7 mg imazapyr a.e./L

48-hr EC50 (?) 373 mg imazapyr a.e./L Eastern oyster EC50 (G) >132 mg imazapyr/L Arsenal® (Crassostrea virginica) NOEC >132 mg imazapyr/L (HDT) Mangels & Ritter 20001 Pink shrimp >132 mg imazapyr/L EC50 (S) (Penaeus duorarum) >132 mg imazapyr/L (HDT) Freshwater water flea AC 243,997 24-hr LC >100 mg imazapyr a.e./L (Daphnia magna) 50 Kintner & Forbis 19832 (technical) 48-hr LC >100 mg imazapyr a.e./L (<24 hours old) 50 AC 243,997 Freshwater water flea 7, 14, 21-day NOEC 97.1 mg/L (HDT, MATC) Manning 19892 (99.5% a.i.) (Daphnia magna) (S/R/G) Grass shrimp AC 243,997 BCF <1 (not calculable) Drotter et al. 19962 (Paleomonetes pugio) (purity NOS) BCF <1 (not calculable) Drotter et al. 19962 AC 243,997 Eastern oyster EC (G) >132 mg/L Drotter et al. 19972 (99.6% purity) (Crassosstrea virginica) 50 AC 243,997 96-hr EC (G) >173 mg/L Ward 19892 (99.5% purity) 50

1 cited in Entrix 10/03 2 cited in SERA 12/04, Appendix 4

Abbreviations: BM = biomass, D = deformity, S = survival; R = reproduction; G = growth; HDT = highest dose tested; MATC = maximum allowable toxicant concentration Table A-12: Toxicity of imazapyr and imazapyr/surfactant mixtures to non-target aquatic vegetation

Test Test Substance Species (Observed Effect) Result Reference 2 Green algae EC50 (G) 71 mg/L Hughes 1987 1 (Selenastrum capricornutum) EC25 (G) 78 mg/L Mangels & Ritter 2000 Freshwater diatom EC50 (G) >59 mg/L Mangels & Ritter 20001 (Navicula pelliculosa) EC25 (G) >59 mg/L Saltwater diatom EC (G) 85 mg/L 50 Hughes 19872 Technical grade (Skeletonema costatum) EC25 (G) 42.2 mg/L imazapyr Blue-green algae EC (G) 117 mg/L 50 Mangels & Ritter 20001 (Anabaena flos-aquae) EC25 (G) 7.3 mg/L Green algae EC (G) 0.2 mg/L Landstein et al. 19932 (Chlorella emersonii) 50 Duckweed EC (G) 0.024 mg/L 50 Hughes 19872 (Lemna gibba) EC25 (G) 0.013 mg/L

EC25 (G shoots) 0.013 mg/L EC50 (G shoots) 0.032 mg/L Common water milfoil EC (# roots) 0.022 mg/L 25 Roshon et al. 19992 (Myriophyllum sibiricum) EC50 (# roots) 0.029 mg/L ® Arsenal + EC25 (G roots) 0.0079 mg/L unidentified EC50 (G roots) 0.0099 mg/L surfactant Green algae EC (G) 14.1 mg/L 50 Mangels & Ritter 20001 (Selenastrum capricornutum) EC25 (G) 8.36 mg/L LC 24 ppb Duckweed 50 Mangels & Ritter 2000 EC (G) 0.0216 mg/L (Lemna gibba) 50 Mangels & Ritter 20001 EC25 (G) 0.0132 mg/L

1 cited in Entrix 10/03. 2 cited in SERA 12/04, Appendix 4. Abbreviations: S = survival; R = reproduction; G = growth; HDT = highest dose tested; MATC = maximum allowable toxicant concentration Table A-13: Toxicity endpoints for risk quotient calculation and levels of concern for interpretation of risk quotients

Aquatic Mammals Birds Aquatic Non-endangered Endangered animals vascular plants plants plants and algae Assessment Acute EC50 or LC50 LD50 oral LD50 oral EC50 EC25 seedling EC25 seedling acute toxicity emergence and emergence and vegetative vigor vegetative vigor or NOEC Chronic NOEC early- NOEC NOEC life stage or full 2-generation 21-week life-cycle tests reproduction reproduction Levels of concern (risk quotient greater than) Acute risk 0.50.5 0.5 1.0 1.0 1.0 Acute restricted use 0.1 0.2 0.2 Acute risk endangered species 0.05 0.1 0.1 Chronic risk 1.0 1.0 1.0

U.S. Environmental Protection Agency, Technical Overview of Ecological Risk Assessment, Analysis Phase: Ecological Effects Characterization and Risk Characterization, September 28th, 2004. APPENDIX 2.

Spartina Control Program Site Maps

All Spartina Control Program Site Maps can be viewed on the Spartina Project’s website at http://www.spartina.org/maps/ssp_maps_2008-10/

Electronic copies on compact disk, or printouts on paper may be obtained for cost of production by calling the Spartina Project Office at 510-548-2461. APPENDIX 3.

Water Quality Field Data Collection Form Page ___ of ___ on this date Field Data Collection Form San Francisco Estuary Invasive Spartina Project, Aquatic Pesticide Application Plan, 2612-A 8th St, Berkeley, CA, 94710

Site ID (XXXX) (eg. CP01): ______Date: ______Collected By: ______

Station Location (circle): at application point upstream downstream Station Type (circle): Reference Treated

Wind (circle): low high Tidal Cycle (circle): high low slack Water Color (circle): green green-brown brown blue (dye)

Herbicide (circle): imazapyr glyphosate Surfactant:______Application Time (Start/Finish): ______/______

Field Measurements Air Temp Water Depth pH Dissolved Water Temp Conductivity Salinity Meter Oxygen Used O C Meters mg/L O C mS ppt

YSI 85

Samples Collected Sample ID (XXXX-YY-Ab)* Time Sample Depth (m) Notes

* XXXX-YY-Ab (eg. DEAD-01-pre, ZIPY-02-0.5h) = XXXX: Site, YY: site visit number (01-1st, 02-2nd, 03-3rd, etc.), A: time to application (either pre, increments thereafter in half hours – 0.5), b: time increment (h=hour, w=week (for 1 week post-treatment))

Additional Notes or Comments: ______

Wildlife presence: ______APPENDIX 4.

Laboratory Chain of Custody Form Analytical Request/Chain of Custody Page _____ of ______Pacific Agricultural Laboratory, LLC PAL Project # ______12505 N.W. Cornell Rd., Ste. 4 • Portland, OR 97229 Tel 503.626.7943 • Fax 503.641.0644 • www.pacaglab.com Pacific Agricultural Laboratory

Name ______Requested Analysis Contact ______Address______City ______State ______Zip ______

Telephone ______Fax ______Requested Turnaround Time Email ______Cell Phone ______

CLIENT INFO Standard (10 working days) Project # ______Purchase Order # ______q Method of Shipment ______q Rush ______please specify

Sample Sample Sample Container No. of PAL ID Client Sample ID Date Time Type Type Containers Comments

DATE TIME DATE TIME Relinquished by: ______Received by ______SIGNATURE SIGNATURE Relinquished by: ______Received by ______SIGNATURE SIGNATURE Lab Comments: ______

All services performed by PAL are subject to the Standard Terms and Conditions on reverse side of this form. Pacific Agricultural Laboratory, LLC Standard Terms and Conditions

Unless otherwise agreed in a separate contract, services provided by Pacific Agricultural Hazardous Materials/Samples: Unused portions of samples found or suspected to Laboratory, LLC (PAL) are expressly limited to the terms and conditions stated herein. be hazardous according to state or federal laws will either be returned to the client at Submission of samples is deemed acceptance of these terms and conditions. the expense of the client, or properly disposed of as hazardous waste at the expense of the client. Samples are the property of the client at all times, even while in the Limits of Liability: All analytical services provided by PAL are made on a best possession or under the control of PAL for analysis. All other samples will be properly effort basis. Established methods of analysis will be followed whenever possible; and anonymously disposed of as nonhazardous waste after expiration of sample retention however every sample has unique properties that may require deviation or adaptation of time. Samples subject to USDA foreign soil or plant permits shall be disposed of in established methodologies. The total liability of PAL will be limited to retesting or refund accordance with applicable permit conditions. of the paid for services provided, at the option of PAL. Use of PAL constitutes acceptance of these limitations of liability. Sample Retention/Disposal: Samples submitted for analysis are retained and stored under proper conditions and security for a period of time after the issuance of the final Confidentiality: Pacific Agricultural Laboratory is zealously protective of customer report. Retention times are generally as follows: information and uses its best business practices to maintain strict and absolute customer confidentiality. Confidentiality can be obtained, to the fullest extent allowed by the law, by placing written, mutual confidentiality agreements into force, upon request. Sample Type Retention Time Information is released to third parties only upon the authorization of the customer, Surface Water, Wastewater 30 days by court order, or otherwise as required by law, and taking precautions to ensure Nonhazardous Soil and Sludge 60 days confidential transfer of information between PAL and its customers by courier or mail, Food Commodities, Plant Tissue 60 days facsimile, email (internet), and/or telephone as the customer may direct. PAL shall not Other Nonhazardous Materials 60 days be responsible for any disclosure of any information of customer unless PAL specifically Hazardous Materials 30 days agrees to keep such information confidential by separate agreement.

Billing: All fees are billed directly to the client. Third party billing will not be accepted Report and Document Retention: PAL shall retain final reports and all supporting without prior arrangement and consent by PAL and agreement of the third party to all documentation and analytical data used to generate reports for eight years following the standard terms and conditions. generation of the report, after which time PAL shall be free to destroy the information.

Payment Terms: For clients without approved credit from PAL, payment must be Sample Containers and Shipping Materials: PAL will provide appropriate sample received prior to the release of final project report. For clients with approved credit, terms containers, shipping containers and packing materials at no additional charge upon prior are net 30 days from the date of invoice unless otherwise stated on that invoice. Any arrangement. Standard shipping will be UPS Ground, and the client will be charged for changes from these terms must be agreed upon prior to sample submission. A service expedited container shipping and the shipping of samples to PAL for analysis. charge of 1 _% per month (18% annual rate) will apply for outstanding balances that are past due. In the event of default of payment for analytical services rendered, the client Analytical Service Requests: Requests for analytical Services can be made by is responsible for reasonable collection charges including court costs and attorney’s fees telephone, fax, email or in writing. The client must confirm requests for service in incurred by PAL. There is an additional $25.00 charge for any returned checks. writing, using a PAL chain of custody form prior to the commencement of work by PAL, and following directions for sampling as provided by PAL. It is very important that the Litigation: All fees and costs associated with compliance by PAL to any subpoena analytical services to be provided by PAL be clearly understood by both PAL and the for documents, for consultation in preparation for or testimony in any deposition or client prior to commencement of projects. PAL will not be responsible for delays caused proceeding, or for any other purpose relating to the work of PAL, in connection with by incomplete information provided by the client including missed hold times and work performed for a client, shall be paid by the client. Such costs include, but are delayed report generation. not limited to, fees for persons responsible for responding to subpoenas, reproduction of reports and data in support of reports, mileage and other travel expenses, attorney preparations for testimony, court testimony, attorney fees, and any other expenses associated with PAL’s participation in the litigation. APPENDIX 5.

Laboratory Quality Assurance Plan 2010

CATALOGUE

Pacific Agricultural Laboratory ultural SpecialistsLa in pesticides bresidue analysiso 12505 N.W. Cornell Rd. Portland, OR 97229-5651 Tel 503.626.7943 Fax 503.641.0644 Pacific Agricultural Laboratory

Specialists in pesticides residue analysis www.pacaglab.com

TABLE OF CONTENTS

Mission Statement ...... 1

Statement of Qualifications...... 2

General Information...... 3

Individual Pesticides and Specialty Analysis ...... 4

Pesticides Screening ...... 13

Specialty Analysis...... 13

EPA Methods Involving Pesticides ...... 16

Other Services ...... 16

PAL Services Catalogue 503.626.7943 MISSION STATEMENT Pacific Agricultural Laboratory is committed to providing the highest quality of analytical services involving laboratory analysis with special emphasis on pesticides and agricultural issues. The laboratory will use the most current methodologies and instrumentation to achieve the highest level of customer satisfaction. Our staff will have the knowledge, experience and training to work effectively with the customer. Pacific Agricultural Laboratory will be an employee-oriented company that provides a personally and professionally satisfying environment in which to work and recognizes the importance of every employee. A comprehensive quality system will assure quality data and the integrity of the results. The laboratory will make every possible effort to meet promised deadlines and work towards providing the highest possible value for the customer. Pacific Agricultural Laboratory is dedicated to creating long term working relationships with our customers regarding analytical needs.

PAL Services Catalogue 503.626.7943 1 STATEMENT OF QUALIFICATIONS Steve Thun, Laboratory Director `ÕV>Ì\ÊÊ °-V°]Ê V iÃÌÀÞ]Ê1ÛiÀÃÌÞÊvÊ"Ài}° Facilities and Instrumentation 9i>ÀÃÊvÊ Ý«iÀiVi\ÊÊ£n Pacific Agricultural Laboratory is located in Portland, Oregon. It occupies 3,500 square feet of laboratory Steve is the owner of Pacific Agricultural Laboratory, space with the primary focus on pesticides residue founded in 1995. Steve oversees all phases of laboratory analysis. Instrumentation dedicated to pesticides operations including administration, quality assurance analysis includes: and client services. Ongoing education has included numerous courses covering gas chromatography, liquid UÊ}iÌÊÈnäÊ}>ÃÊV À>Ì}À>« ÊiµÕ««i`ÊÜÌ Ê>Ê chromatography, extraction techniques including 5973 quadrapole mass spectrometer. supercritical fluid extraction, and management training. UÊ}iÌÊÈnäÊ}>ÃÊV À>Ì}À>« ÊiµÕ««i`ÊÜÌ Ê`Õ>Ê Steve previously worked as Senior Chemist in an electron capture detectors. export laboratory implementing an extensive pesticides UÊ««i`Ê ÃÞÃÌiÃÊ*ÊÓäääÊ* ÌÀ«iÊµÕ>`Ê -Ê residue program in compliance with ISO 9002 quality ÃÞÃÌi°Ê µÕ««i`ÊÜÌ Ê>Ê}iÌÊ££ääÊ* Ê>`Ê>Ê guidelines. triple quadrupole mass spectrometer. UÊ}iÌÊ££ääÊ* -ÊÃÞÃÌi°Ê µÕ««i`ÊÜÌ Ê>Ê Richard Jordan, Laboratory Manager UV-Vis diode array detector and a quadrupole mass `ÕV>Ì\ÊÊ °-V°]Ê* ÞÃVÃ]Ê7>Ã }ÌÊ-Ì>ÌiÊ1ÛiÀÃÌÞ° spectrometer. 9i>ÀÃÊvÊ Ý«iÀiVi\ÊÊ£n UÊ}iÌÊ££ääÊ* vÕÀiÃViViÊ`iÌiVÌÀÊÃÞÃÌi°Ê Equipped with a Pickering post column reactor and Rick has been actively involved with analytical fluorescence detector configured for Glyphosate analy- chemistry for the past eighteen years. Rick has been sis. involved with every phase of semivolatile organic UÊiÜiÌÌÊ*>V>À`Ê£äxäÊÃÞÃÌiÊVv}ÕÀi`ÊvÀÊ}iÊ«iÀ- analysis with extensive gas chromatography experience. meation chromatography. Rick comes to Pacific Agricultural Laboratory via UÊiÜiÌÌÊ*>V>À`ÊxnäÊÊ}>ÃÊV À>Ì}À>« ÊiµÕ««i`Ê Test America (formerly North Creek Analytical) and with flame photometric detector (FPD) and flame Oregon Analytical Laboratory, where as Senior Chemist, ionization (FID) detector. he was involved with managing the semivolatiles department. Rick is responsible for laboratory UÊ*ÀÕÊ iiÌÊ>LÀ>ÌÀÞÊvÀ>ÌÊ>>}i- operations, sample workload scheduling, pesticides ment system (LIM). analysis and instrumentation. Rick’s specialties are gas Personnel chromatography and mass spectrometry. Acquiring and retaining highly qualified and motivated people within Pacific Agricultural Laboratory is the key to our success. Our key personnel have 51 years of experience directly involved with pesticides analysis and analytical chemistry with an average of over ten years per person. Ongoing training, membership in various professional associations and an active involvement in continuing education essential to keeping pace with the rapid pace of technology are critical to maintaining the highest level of staff qualification.

PAL Services Catalogue 503.626.7943 2 QUALITY ASSURANCE ANALYSIS REPORTING AND Pacific Agricultural Laboratory operates under a TURNAROUND comprehensive quality system that covers all aspects The timely delivery of analytical results is an integral of laboratory operations. This quality system is part of Pacific Agricultural Laboratory’s mission. All documented in the Pacific Agricultural Laboratory results are reported via facsimile or email upon final Quality System Manual. The quality system is designed completion of analysis. Final reports are mailed on to meet all quality standards set forth by government or before the next working day. All results released to organizations such as the EPA, private accrediting clients are treated as confidential information, and no L`iÃÊ>`ÊÌ iÊViÌÃÊÌ iÃiÛiÃ°Ê7iÊ >ÛiÊ`iÃ}i`ÊÌ iÊ results will be released to any third party without the quality system to be flexible yet comprehensive enough consent of the client. to meet customer needs, whatever they may be. Copies of the Quality System Manual will be provided upon Standard Turnaround request. 10 business days List price The value of service provided by Pacific Agricultural Laboratory is measured in the accuracy and precision of Rush Services* the results that we provide to our clients. To maintain 5 Day Rush a consistently high standard of analytical excellence, 5 business days List price + 40% Pacific Agricultural Laboratory participates in various check sample programs, and makes the results of those 48 Hour Rush studies available to clients for inspection upon request. ÓÊLÕÃiÃÃÊ`>ÞÃÊ ÃÌÊ«ÀViÊ³Ênä¯ ACCREDITATION, CERTIFICATIONS * Rush service is deemed complete upon transmission of verbal, email and/or facsimile results. Weekend rush service AND MEMBERSHIPS is available with prior arrangement. UÊ iÀÌvi`Ê>LÀ>ÌÀÞ]Ê7>Ã }ÌÊ i«>ÀÌiÌÊvÊ V}Þ]Ê>LÊVVÀi`Ì>ÌÊ £nxn PRICING STRUCTURE AND TERMS UÊ1- ÊÀi}Ê-Ê*iÀÌÊ*ÎÎäänää£Çx Standard terms are net 30 days with credit approval. Pacific Agricultural Laboratory may require payment UÊ1- ÊÀi}Ê*>ÌÊ*iÀÌÊ* *änääÓÓn before release of analytical results. Accounts over 30 UÊAssociation of Official Analytical Chemists (AOAC), days are subject to 1.5% per month service charge. member Discounts may apply to large sample lots or contract work on a prearranged basis. Please inquire about UÊAmerican Chemical Society, member pricing for services of interest not listed in the catalog.

UÊ7iÃÌiÀÊ7>Ã }ÌÊvÊ ÕÀÃiÊ-Õ«iÀÌi`iÌÃÊ Association, member LIMITS OF LIABILITY

UÊ"Ài}Ê ÕÃiÃÃÊÃÃV>Ì]ÊiLiÀ All analytical services provided by Pacific Agricultural Laboratory are made on a best effort basis. Established UÊ7>iÌÌiÊ6>iÞÊ}Ê*ÀviÃÃ>ÃÊÃÃV>Ì methods of analysis will be followed whenever possible, however every sample has unique properties that may require deviation or adaptation of established methodologies. The total liability of Pacific Agricultural Laboratory will be limited to the invoice amount for services provided. Use of Pacific Agricultural Laboratory constitutes acceptance of these limits.

PAL Services Catalogue 503.626.7943 3 PESTICIDES SCREENING organonitrogen and n-methyl carbamates. It is designed to comply with Oregon DEQ Guidance for Evaluating Food and Agricultural Commodities Residual Pesticides on Lands Formerly Used for Agricultural Production (DEQ-06-LQ-011) and for Pacific Agricultural Laboratory uses the FDA Pesticide other environmental screening purposes. Please inquire Analytical Manual–Volume I as the primary reference for current compound lists. for pesticides screening for food products and animal feed. The methodologies are comprised of Halogenated Pesticides multiresidue methods. The actual number and identity Residue Screen...... $160.00 of determinable residues is dependent on the sample Pacific Agricultural Laboratory offers an extensive screen matrix and the number and variety of determinative for halogenated pesticides in water, soil and agricultural steps involved. In addition to multiresidue methods commodities. Analysis is done by gas chromatography listed in the Pesticide Analytical Manual, Pacific using electron capture detection (ECD). Agricultural Laboratory also provides custom screens based on customer needs. Method reporting limits vary Acetochlor depending on the matrix and may be raised due to Alachlor interference of coextracted compounds. Aldrin ivÕÀ> SOIL, WATER AND OTHER _ ` MATRICES b a Ê`>i® iÃìÃÊv`ÃÊ>`Ê>}ÀVÕÌÕÀ>ÊV`ÌiÃ]Ê*>VvVÊ viÌ À Agricultural Laboratory tests for pesticides in soil, Captafol water, chemical formulations and other matrices. Captan Methodologies come from a variety of sources such as Chlordane the U.S. EPA, trade journals, manufacturers and in- Chloroneb (Terraneb) house method development. Chlorpyrifos (Lorsban) Chlorpyrifos-methyl SCREENS PROVIDED BY PACIFIC Chlorobenzilate AGRICULTURAL LABORATORY ÀÌ >Ê À>Û® Cyfluthrin Multiresidue Screen in Foods Cyhalothrin and Agricultural Commodities ...... $330.00 Cypermethrin Dacthal (DCPA) This screen is based on both FDA Pesticide Analytical DDD Manual Method 302 (Luke extraction) and The DDE California Department of Food and Agriculture Multi- DDT ,iÃ`ÕiÊ-VÀiiÊ£nn®°ÊÌÊVÕìÃÊÛiÀÊ£ÇäÊ«iÃÌVìÃÊ Deltamethrin comprised of organochlorine, organophosphorous, VÀ>Ê ÌÀ>® organonitrogen and n-methyl carbamates. Please inquire Dicofol (Kelthane) for specific compound lists and compatibility with your Dieldrin specific commodity. Dichlobenil (Casoron) Multiresidue Screen in Soil and Water .. $330.00 Endosulfan I(Thiodan) Endosulfan II This screen is a combination of four Modified EPA Endosulfan sulfate iÌ `Ã\ÊÊ *Ên£{£ Ê * ®]Ê *Ênän£ Ê Endrin

®]Ê *ÊnÓÇä Ê -]Ê- Êì®Ê>`Ê *Ê Endrin aldehyde nÎÓ£ Ê* -®°ÊÌÊVÕìÃÊÛiÀÊ£ÇäÊ«iÃÌVìÃÊ Endrin ketone comprised of organochlorine, organophosphorous,

PAL Services Catalogue 503.626.7943 13 Esfenvalerate (Asana) Dithiocarbamate Fungicides Screen*..... $140.00 Ethalfluralin (Sonalan) Mancozeb (Dithane) Fenarimol (Rubigan) Maneb Fenvalerate Nabam Flutonanil (Prostar) Thiram Folpet Vapam (Metam-sodium) i«Ì>V À Zineb i«Ì>V ÀÊi«Ý`i Ziram iÝ>V ÀLiâii Iprodione (Rovral) Metolachlor * This analysis is a presumptive test for the presence Methoxychlor of thiocarbamate pesticides. This method does not Mirex discriminate between individual thiocarbamate Norflurazon (Solicam) pesticides. Ovex Oxadiazon (Ronstar) Triazine Herbicides Screen ...... $160.00 Oxyfluorfen (Goal) Ametryn PCA Atrazine * Þ>>âiÊ >`iÝ® Pendimethalin iÝ>âiÊ6i«>À® Permethrin Metribuzin (Sencor) Prodiamine Prometon (Pramitol) Pronamide (Kerb) Prometryn Propachlor Propazine Propanil Simazine Propiconazole (Tilt) Simetryn Terbacil Terrazole Phenylurea Herbicides Screen ...... $160.00 Triflumizole Chlorpropham (CIPC) Trifluralin Diuron Toxaphene DCPMU (degradate of Diuron) Vinclozalin (Ronilan) Fenuron Linuron N-methyl Carbamate Pesticides Monuron Residue Screen...... $180.00 Neburon Aldicarb (Temik) Propham (IPC) Aldicarb Sulfone Siduron i`V>ÀL Carbaryl (Sevin) Carbofuran (Furadan) ÎÞ`ÀÝÞV>ÀLvÕÀ> iLÕV>ÀLÊ >ÞV>ÀL® Methiocarb (Mesurol) Methomyl (Lannate) Oxamyl (Vydate) *À«ÝÕÀÊ >Þ}® Thiobencarb

PAL Services Catalogue 503.626.7943 14 Organophosphorous Pesticides Chlorinated Acid Herbicides Screen Residue Screen...... $160.00 Water...... $185.00 Aspon Soil and Plant...... $200.00 Azinphos-methyl 2,4-D ÃÌ>ÀÊ-Õ«ÀvÃ® Ó]{ Carbofenothion 2,4-DP (Dichlorprop) Chlorpyrifos (Lorsban) 2,4,5-T Chlorpyrifos-methyl (Reldan) 2,4,5-TP (Silvex) Coumaphos Acifluorfen Demeton (Systox) iÌ>â Demeton-methyl (Metasystox) Clopyralid (Stinger) Diazinon V>L>Ê >Ûi® Dichlofenthion Dinoseb (Dinitro) Dichlorvos (DDVP) MCPA Dicrotophos MCPP Dimethoate Pentachlorophenol Disulfoton Picloram (Tordon) EPN Quinclorac Ethion Triclopyr (Garlon) Ethoprop (Mocap) Famphur Fenitrothion Specialty Analysis Fensulfothion UÊForeign Soil and Plant Tissue Analysis Fenthion UÊGlyphosate (Roundup) Fonophos UÊImidazolinone herbicides Malathion UÊFormulation checks Mevinphos UÊ-ÕvÞÕÀi>ÊiÀLV`iÃ Monocrotophos Naled Parathion Parathion methyl Phorate Phosmet (Imidan) Phosphamidon Tetrachlorvinphos Terbufos Trichlorfon (as Dichlorvos)

PAL Services Catalogue 503.626.7943 15 EPA METHODS INVOLVING PESTICIDES

Analytes EPA Method Price

Þ« Ã>ÌiÊÊ7>ÌiÀ 547 $160.00

µÕ>ÌÊ>`Ê*>À>µÕ>ÌÊÊ7>ÌiÀ 549.1 f£nä°ää

Organochlorine Pesticides Èän]Ênän£ $150.00

Organophosphorous Pesticides È£{]Ên£{£ $175.00

Ì V>ÀL>>ÌiÊ*iÃÌV`iÃÊÊ7>ÌiÀ 630.1 $140.00

iÞÊ>`Ê >ÀLi`>âÊÊ7>ÌiÀ 631 $160.00

>ÀL>>ÌiÊ>`Ê1Ài>Ê*iÃÌV`iÃÊÊ7>ÌiÀ ÈÎÓ]ÊnÎÓ£ $170.00

À>Ìi`ÊiÀLV`iÃ È£x]Ên£x£]ÊnÎÓ£ f£nx°ää

Miscellaneous Pesticides nÓÇä Ê -]Ê- Ê`i® Per bid

N-Methyl Carbamate Pesticides nÎÓ£ $170.00

ÃVi>iÕÃÊ*iÃÌV`iÃÊ* -® nÎÓ£ Per bid

Additional methods are being validated and added on a regular basis. If there is an analytical method that is required but not listed, please inquire. Other EPA methods not specifically involving pesticides are also available.

OTHER SERVICES OTHER SERVICES PROVIDED Pacific Agricultural Laboratory offers a variety of other UÊGC-MS forensic analysis analytical services not relating specifically to pesticides UÊMetals or foods. These services may be done in-house or UÊMicrobiology subcontracted to approved subcontract laboratories. UÊGeneral and wet chemistry Through networking with other laboratories, Pacific UÊEnvironmental testing Agricultural Laboratory can ease the problem of finding multiple sources of analysis for a particular project. CONSULTING SERVICES Consulting services related to pesticides, agricultural products and foods are available for special projects, sample collection, data review and validation. Consulting is done on an hourly basis.

PAL Services Catalogue 503.626.7943 16 QUALITY SYSTEM MANUAL Revised 7/10/06 Table of Contents

QUALITY POLICY 1

1.0 COMPANY DESCRIPTION 2

2.0 ETHICS POLICY 2

3.0 QUALITY SYSTEM MANAGEMENT 3

4.0 QUALITY ASSURANCE/QUALITY CONTROL POLICY 9

5.0 QUALITY ASSURANCE PROCEDURES 10

6.0 STANDARD OPERATING PROCEDURES 12

7.0 CORRECTIVE ACTION 13

8.0 CUSTOMER COMPLAINTS 14

9.0 DOCUMENT CONTROL 14

10.0 SYSTEM AND PERFORMANCE AUDITS 18

11.0 DATA QUALITY OBJECTIVES 18

12.0 CHEMICALS, REAGENTS, AND ANALYTICAL STANDARDS 20

13.0 EQUIPMENT 20

14.0 SAMPLE RECEIPT AND LOG-IN PROCEDURES 22

15.0 ANALYTICAL PROCEDURES 23

16.0 SAMPLE RETENTION AND DISPOSAL 23

17.0 COMPUTERS 24

18.0 LABORATORY PERSONNEL TRAINING 24

19.0 SAFETY 25

20.0 ENVIRONMENTAL POLICY 25

21.0 SUBCONTRACTING OF SERVICES 26

APPENDIX A ANALYTICAL INSTRUMENTATION LIST 27 Quality System Manual Pacific Agricultural Laboratory

QUALITY POLICY

Pacific Agricultural Laboratory is committed to providing the highest quality of analytical services attainable regarding pesticide analysis and agricultural issues. Emphasis on quality is underscored by the total commitment of the management to good laboratory practice and our quality assurance program. Pacific Agricultural Laboratory recognizes that meeting its quality standards is dependent on the collective contribution of all employees, which must have the proper training, knowledge, and experience to meet the ongoing company objective of complete customer satisfaction. This commitment is the foundation of a high quality testing service that allows us to provide customers with accurate, meaningful, and timely information.

Steve Thun Laboratory Director

1 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory

1.0 COMPANY DESCRIPTION

1.1 Pacific Agricultural Laboratory was established in June, 1995 and is located in Portland, Oregon.

1.2 Pacific Agricultural Laboratory performs pesticide analysis in a variety of sample matrices including water, soil, plant tissue, and product formulations. Analyses are performed on a single analyte, targeted screening, or multiresidue profile basis.

1.3 Pacific Agricultural Laboratory also facilitates other agricultural and environmental testing services. These analyses are performed by sub-contractors at certified laboratories.

1.4 Laboratory access is restricted to authorized personnel. Visitors are welcome but must be accompanied by a laboratory employee and must observe all laboratory safety practices.

2.0 ETHICS POLICY

2.1 Pacific Agricultural Laboratory is a customer-driven company and welcomes competition. The core of the Pacific Agricultural Laboratory ethics policy is integrity. Services to customers are provided in a clear, accurate, truthful, and timely fashion. False or artificial entries into company records are not tolerated for any reason.

2.2 Approved procedures for all aspects of company operations are conformed to at all times. Deviations from approved procedures must be approved by management and must be documented. Pacific Agricultural Laboratory does not deviate from contractual requirements unless deviations are approved by the customer.

2.3 Pacific Agricultural Laboratory is zealously protective of customer information and maintains strict and absolute customer confidentiality. Confidentiality is maintained according to SOP QA-012 in the following manner.

2.3.1 Written, mutual confidentiality agreements are placed into force upon the request of the customer.

2.3.2 Analytical results are released to third parties only upon the express authorization of the customer.

2 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 2.3.3 Precautions are taken to ensure confidential transfer of information between Pacific Agricultural Laboratory and its customers. This includes the transmission of information by facsimile, and telephone.

2.4 Pacific Agricultural Laboratory is highly vigilant at all times to avoid conflicts of interest. Accordingly, gifts (including monetary) from vendors, suppliers, or contractors cannot be accepted by Pacific Agricultural Laboratory employees. In the event that a gift arrives at the company unannounced, the gift will be returned to sender.

Pacific Agricultural Laboratory is committed to following all stated procedures as written (see 2.2). Requests from customers to expedite the acquisition of results at the expense of deviating from procedures, or requests to alter results will not be accommodated. All incidents of this nature are reported directly to the Laboratory Director.

2.5 Pacific Agricultural Laboratory places grave importance on this ethics policy. Non-compliance with this policy will result in immediate disciplinary action up to, and including termination.

3.0 QUALITY SYSTEM MANAGEMENT

3.1 The success of Pacific Agricultural Laboratory Quality System depends on the cooperation of the entire Pacific Agricultural Laboratory staff. All employees contribute to the production of quality analytical data by understanding the Quality System and by implementing and respecting the associated QA/QC procedures.

3.2 Responsibility for the management of the Quality System is distributed between the Laboratory Director, the Quality Manager, and the Laboratory Manager. Routine daily operations that contribute to the Quality System are performed by all individuals at Pacific Agricultural Laboratory. The relationship between these individuals is defined in the accompanying organizational chart. Individual responsibilities are described below.

3.2.1 Laboratory Director:

Ultimate responsibility for the implementation and monitoring of the Quality System rests with the Laboratory Director of Pacific Agricultural Laboratory who is the only approved signatory. Specific responsibilities include:

3.2.1.1 Approval and implementation of the Quality System and Standard Operating Procedures (SOPs).

3 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 3.2.1.2 Delegation of Quality System responsibilities and ensuring that the company remains in compliance with the requirements of the Quality System.

3.2.1.3 Insure Chemical Hygiene Plan is Followed. Serve as the Chemical Hygiene Officer. Perform annual review of the Chemical Hygeniene Plan.

3.2.1.4 Provision of sufficient personnel and resources for normal lab operations and for special projects.

3.2.1.5 Notification of staff regarding customer-specific contract requirements.

3.2.1.6 Maintenance of appropriate certification currency.

3.2.1.8 Customer complaints resolution.

3.2.1.9 Document change control.

3.2.1.10 Secure maintenance of documentation in an organized and systematized Master File.

3.2.1.11 Review and approval of raw data prior to the completion of Data Analysis Report Forms.

3.2.1.12 Review and approval of method quality control prior to release of data for entry into final reports.

3.2.1.13 Development and maintenance of electronic databases and database security.

3.2.2 Quality Manager:

3.2.2.1 Monitoring of control charts, logbook entries, and document storage.

3.2.2.2 Development of SOPs with the assistance of relevant participants and verification that SOPs are implemented, properly executed and complied with.

3.2.2.3 Quality System audits and the filing of written annual reports on the status of the Quality System.

3.2.3 Laboratory Manager:

4 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 3.2.3.1 Oversight of the training of new employees.

3.2.3.2 Maintenance of appropriate sample hold/turnaround times and communication of delays to customers.

3.2.3.3 Full understanding of all technical methods used in the lab.

3.2.3.4 Assignment of individual most appropriate to perform analytical procedure(s) on sample(s).

3.2.3.5 Documentation of equipment maintenance, malfunctions, and repairs.

3.2.3.6 Method selection

3.2.3.7 Purchase and receipt of equipment and supplies. Monitoring of supply and reagent inventories.

3.2.3.8 Correction of out-of-control situations and documentation of corrective actions as per SOP QA 004.

3.2.3.9 Maintenance of accident and spills file as per SOP QA-009.

3.2.4 Office Manager:

3.2.4.1 Assist the Laboratory Director with general administrative duties not related to the generation, interpretation, or reporting of laboratory data.

3.2.4.2 Customer communications.

3.2.4.3 Bookkeeping and Accounting

3.2.4.4 Shipping and receiving.

3.2.4.5 Assistance with additional administrative duties defined by the Laboratory Director.

3.2.5 Marketing Manager:

3.2.5.1 Develop new customer contacts.

3.2.5.2 Advertising and promotion of Pacific Agricultural Laboratory.

3.2.5.3 Promote and maintain good customer relations.

5 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 3.2.6 Laboratory Technician:

3.2.6.1 Execution of methods under the supervision of the Laboratory Manager.

3.2.6.2 Conform to quality control procedures described in the Quality System Manual and method-specific SOPs.

3.2.6.3 Completion of method worksheets, including worksheet calculations.

3.2.6.4 Participation in the revision of existing SOPs and the drafting of new SOPs

3.2.6.5 Identification of out of control situations and reporting same to Laboratory Manager.

3.2.6.6 Routine and non-routine equipment maintenance.

3.2.6.7 Calibration and maintenance of laboratory analytical balances and pH meters.

3.2.7 Laboratory Analyst:

3.2.7.1 Instrument calibration

3.2.7.2 Execution of instrumental analysis methods under the supervision of the Laboratory Manager.

3.2.7.3 Routine and non-routine equipment maintenance.

3.2.8 Sample Custodian:

3.2.8.1 Under the supervision of the Laboratory Manager, proper sample log-in, handling and storage.

3.2.8.2 Assignment of Pacific Agricultural Laboratory identification number.

3.2.8.3 Safe and proper disposal of samples including provisions defined in USDA foreign soil import permit.

3.2.8.4 Monitor and maintain sample storage areas including sample refrigerators and freezers.

6 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory Pacific Agricultural Laboratory Organizational Chart

7 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 3.3 Deputies

3.3.1 The Laboratory Manager will assume the responsibilities (including signatory) of the Laboratory Director during his/her absence.

3.3.2 The responsibilities of the Laboratory Manager will be reassigned by the Laboratory Director in the absence of the Laboratory Manager.

3.3.3 No deputies exist for the remaining positions within the company at this time.

3.4 Communication

3.4.1 Daily

Analytical data are evaluated by the Laboratory Analyst for precision, accuracy, and completeness. Data sets and packages are reviewed and inspected for completeness before the results are released to the customer.

3.4.2 Annually

A written annual report of a Quality System audit will be prepared that describes laboratory performance, Quality System problems and their respective corrective actions, laboratory audits, and training. The report will also outline the status of SOPs, the Chemical Hygiene Plan, safety practice, and certification.

3.5 Quality System Assessment and Review

3.5.1. The effectiveness of the Quality System is monitored on a continuous basis and reviewed in detail on a annual basis.

3.5.2 The Quality System Manual and SOPs are formally reviewed annually as a general laboratory policy. Reviews may be initiated by analytical staff whenever warranted, e.g., by a change in instrumentation or the implementation of an improved method.

3.6 Laboratory Environment

3.6.1 Adequate lighting, energy sources, temperature control, and ventilation is maintained to optimize proper performance of analytical procedures.

3.6.2 The laboratory is configured to prevent the invalidation of results or the generation of inaccurate measurements.

3.6.3 The laboratory is maintained in a clean, orderly, and systematized fashion.

8 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory

3.6.4 Adequate space is dedicated to allow for the proper execution of laboratory tasks including bench work, calculations, and report writing.

3.6.5 The laboratory work area is defined and controlled from unauthorized access.

3.6.6 Routine housekeeping measures include regular cleaning and dusting of equipment, floors, and all work surfaces.

4.0 QUALITY ASSURANCE/QUALITY CONTROL POLICY

4.1 The purpose of Quality Assurance/Quality Control (QA/QC) activities at Pacific Agricultural Laboratory is to achieve and maintain high quality laboratory performance.

4.2 The success of the Pacific Agricultural Laboratory Quality System is directly dependent on employee interaction with management and the direct input of employees into QA/QC procedures. The Quality System policies and practices described in this manual are available to employees at all times; and comments, suggestions, and input from employees regarding the Quality System are encouraged.

4.3 Quality Assurance is a set of practices designed to ensure that data generated in the laboratory is of known and adequate quality. Key elements of Pacific Agricultural Laboratory’s Quality Assurance include the use of approved analytical methods, equipment maintenance, sample and document control, reference standards, audits, corrective actions, a mechanism to address customer complaints, and laboratory training practices.

4.4 Quality Control activities include day-to-day monitoring to verify the quality of laboratory operations and procedures. These practices include instrument calibration, observance of good laboratory practice, adherence to standard operating procedures and structured documentation, and proper analysis and documentation of testing results.

4.5 Pacific Agricultural Laboratory’s Quality System has been designed to meet the following objectives:

4.5.1 Follow good laboratory practice.

4.5.2 Maintain high quality laboratory performance.

4.5.3 Maintain rigorous document control.

9 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 4.5.4 Identify and implement high quality analytical methods and practices.

4.5.5 Determine method ruggedness and establish method control limits based on in-house validation studies.

4.5.6 Create a record of instrument performance to support method validation and instrument trouble-shooting.

4.5.7 Detect training needs.

4.5.8 Continuously monitor and assess the quality of laboratory practice.

4.5.9 Execute laboratory operations in a safe and an environmentally sensitive manner.

4.6 Recognizing the importance of QA and QC procedures in laboratory operations, the Laboratory Director of Pacific Agricultural Laboratory is dedicated to the implementation, execution, and monitoring of the Quality System. In addition, the Laboratory Director is committed to ensuring that the policies and practices of this Quality System manual are communicated to, and understood by all employees.

5.0 QUALITY ASSURANCE PROCEDURES

5.1 Pacific Agricultural Laboratory’s QA procedures are a collection of practices and procedures designed to prevent and/or detect analytical error. The ultimate objective of these procedures is to ensure the quality and reliability of all data reported. The following procedures enable Pacific Agricultural Laboratory to generate data of consistent accuracy and precision. Specific QC requirements for each method are described in Pacific Agricultural Laboratory SOPs.

5.2 Internal QA Checks

5.2.1 Laboratory control samples are used to confirm that the laboratory remains in control throughout the analytical process. Control samples consist of the appropriate reference material with known amounts of the characteristic being measured on the unknown sample. Control samples are handled in the same way as an unknown sample throughout all phases of the analytical procedure.

5.2.2 Upper and lower control limits are established for analytes, and are unique for each method. Method specific control limits are defined in method SOPs and/or kept in a control limit database that is available for analysts

10 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory to refer to. Control limits are determined following statistical analysis of historical databases and in-house validation studies. If control limits for a specific method are exceeded, method specific corrective action will be taken, which may include re-testing.

5.3 Calibration Procedures and Frequency

5.3.1 Instrument calibration methods described in Pacific Agricultural Laboratory laboratory SOPs. The frequency of calibration is instrument- specific and is based on manufacturer recommendations, industry requirements, and the analyst’s professional judgment.

5.3.2 Calibration standards are either obtained from traceable commercial sources, or are prepared from high quality commercial materials. Commercial calibration standards have accompanying traceability documentation verifying performance characteristics. Schedules for factory re-calibration of specific standards (e.g., balance standard, oven thermometers) are defined on equipment maintenance sheets. Calibration SOPs describe the identity and proper use of these standards during the calibration process. The performance characteristics of standards prepared from commercial materials are verified by inter comparison with existing laboratory standards before they are accepted for use in laboratory analysis. Calibration standards are used for calibration only and for no other purpose.

5.3.3 All instrument calibrations, corrective actions, and maintenance activities are documented in an Equipment log book specific to each instrument, which is kept in the laboratory.

5.4 Preventative Maintenance

5.4.1 Aggressive and proactive instrument monitoring is a Pacific Agricultural Laboratory policy designed to prevent instrument failure, minimize down time, and avoid interruptions in the normal flow of laboratory operations. Analysts are trained in all aspects of routine instrument operation and maintenance.

5.4.2 Whenever possible, service contracts with the instrument vendors are maintained on an on-call basis. Should instrument down-time occur, the vendor contract stipulates the response time for service calls. Instrument repairs may also be performed by Pacific Agricultural Laboratory technical staff.

5.4.3 In the event that an instrument is deemed unfit for service, the instrument is labeled as being out of order and is removed and stored outside the laboratory until it has been repaired.

11 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory

5.4.4 Laboratory SOPs provide detailed descriptions of maintenance procedures. Instrument maintenance documentation is maintained in a log book which contains a complete record of all service and maintenance, including details of repairs. Additional repair detail may also be found in Corrective Action Reports.

5.5 Data Validation

5.5.1 All sample preparation and sample analyses are documented on method worksheets specific for the analytical procedure. Documentation includes, but is not limited to, Pacific Agricultural Laboratory identification number, sample log-in date, sample description, analytical method(s), analyst, analysis results, and test date. Raw analytical data and calculations leading to final results are recorded on method specific worksheets.

5.5.2 Analytical results are reviewed at two levels before they are released to the customer.

5.5.2.1 The Laboratory Technician and/or the Laboratory Analyst reviews his/her own data to confirm that sample preparation and analytical procedures were executed properly, and that documented information is accurate. The Laboratory Technician is also responsible for confirming that methodology conforms to standard operating procedures, and that all calculations are complete and accurate.

5.5.2.2 Data and analytical QA are reviewed and approved by the Laboratory Director prior to release to the customer.

6.0 STANDARD OPERATING PROCEDURES (SOPs)

6.1 The proper procedure for writing SOPs is described in SOP QA-001

6.2 All SOPs and SOP revisions will conform as closely as possible to the following format:

6.2.1 SOP Number and Title

6.2.2 Scope and Application

6.2.3 Summary of Method

6.2.4 Interferences

12 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory

6.2.5 Sample Handling and Preservation

6.2.6 Apparatus and Instrumentation

6.2.7 Reagents and Supplies

6.2.8 Procedures

6.2.9 Calculations

6.2.10 Quality Control

6.2.11 Reporting

6.2.12 References

6.2.13 Figures and Appendices

6.3 SOPs are reviewed and authorized prior to implementation.

6.4 Deviations from approved procedures must be approved and documented.

6.5 A Document Action Form must be completed and authorized prior to incorporating revisions into an existing SOP.

6.6 SOPs are systematically reviewed for currency and completeness.

6.7 SOPs are listed by number and title in the Master File document F009.

7.0 CORRECTIVE ACTION

7.1 Corrective action is undertaken when errors, deficiencies, or out-of-control situations are identified. The corrective action procedure is described in SOP QA-004 and is documented by the completion of a “Corrective Action Form” by the Quality Manager

7.2 Corrective actions are often initiated by the analyst following the routine monitoring of preparative and analytical procedures, instrument performance, calculations, transcription, review of raw data, and preparation of Materials Analysis Report Forms.

7.3 The corrective action process begins when sporadic or persistent problems are communicated to the Laboratory Manager. At this point in the process, sample analysis is discontinued until the reported problem(s) has been resolved. If data

13 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory integrity for samples previously tested are suspect, the samples are re-analyzed once the appropriate corrective action has been implemented.

7.4 If an erroneous report has been issued to the customer, the customer is contacted immediately by telephone and a corrected written report is issued to the customer as soon as it becomes available.

7.5 Corrective Action Forms are kept in a Corrective Action File and used to assist in future trouble shooting episodes.

8.0 CUSTOMER COMPLAINTS

8.1 Pacific Agricultural Laboratory is customer-driven and committed to providing customers with the highest quality product possible. The Company places top priority on resolving customer complaints in the most expeditious fashion possible. The mechanism for addressing customer complaints is defined in SOP QA-003 and consists of the following:

8.1.1 A detailed account of the complaint is documented on a Customer Complaint Form.

8.1.2 The Laboratory Director is notified immediately and provided with a detailed description of the complaint.

8.1.3 The method specific worksheets supporting the final report provided to the customer are reviewed for proper method execution, calculations, and the accuracy of final values.

8.1.4 The customer is notified telephonically or in writing of the findings of the complaint investigation as soon as the investigation is complete.

8.2 The Customer Complaint Form is entered into the Customer Complaint file.

8.3 If the investigation of the complaint reveals that an error was made leading to the provision of incorrect information to the customer, the sample(s) is re-analyzed and the results are forwarded to the customer. If appropriate, corrective action to prevent the recurrence of the problem is taken and documented.

9.0 DOCUMENT CONTROL

9.1 All documentation associated with laboratory procedures and analysis of samples is maintained in an organized Quality System. Components of the quality system include the Quality Sytem Manual, Chemical Hygiene Plan, standard operating

14 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory procedures and laboratory forms. All components of the quality system are listed in the PAL Quality System Master File List, form F009.

9.2 Document control begins with the receipt of the customer sample. Upon receipt, the contents of the shipping container are examined to confirm that the contents are consistent with the attendant shipping information (or telephonic communication).

9.3 Samples are logged in by completing a Sample Log-in Form as per SOP QA- 002, and each sample is assigned a unique Pacific Agricultural Laboratory identification number in the order which it is received.

9.4 The unique Pacific Agricultural Laboratory identifier accompanies the sample and all attendant documentation throughout the completion of the analytical process. The documents that are linked by the Pacific Agricultural Laboratory identification number are as follows:

9.4.1 Sample Log-in Forms

9.4.2 Sample Extraction Worksheets

9.4.3 Sample Data Analysis Worksheets

9.4.4 Final Reports

9.4.5 Corrective Action Forms

9.4.6 Customer Complaint Forms

9.4.7 Electronic documents

9.5 Documentation per se consists of the following:

9.5.1 Sample log-in forms

9.5.2 Equipment data sheets

9.5.3 Equipment maintenance sheets

9.5.4 Equipment calibration records

9.5.5 Vendor and supplier file

9.5.6 SOPs

15 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 9.5.7 Material testing worksheets

9.5.8 Materials analysis report forms

9.5.9 Customer complaint forms

9.5.10 Corrective action forms

9.5.11 Accident/spill Forms

9.5.12 Training forms

9.5.13 Materials received forms

9.5.14 Material purchased forms

9.5.15 Document action forms

9.5.16 Electronic databases

9.5.16.1. Final Customer reports

9.5.16.2 Analytical Control Charts

9.5.16.3 PAL Quality System

9.6 Document Security

9.6.1 Printed, handwritten or electronic documentation pertaining specifically to the analysis of customer samples is maintained on the Pacific Agricultural Laboratory premises for a minimum of ten years, or longer if designated by regulatory requirements.

9.6.2 All printed documentation in the Pacific Agricultural Laboratory Quality System is maintained indefinitely.

9.6.3 Electronic data and customer reports are maintained for a minimum of ten years.

9.6.4 Access to documentation is restricted to authorized Pacific Agricultural Laboratory personnel.

9.6.5 Security of electronic data is maintained using a back up procedure described in SOP QA-005.

16 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 9.7 Document Change Control System

9.7.1 All documentation is protected by a document change control system. The system ensures that only current and appropriate documents are available in the laboratory. In addition, the system prevents the mistaken use of obsolete documents and provides a system for retention of obsolete documents for the purpose of knowledge-preservation. If a document requires revision, or if an existing document requires removal from the Master File, or if a new document is to be added to the Master File, the following document change control system is engaged.

9.7.1.1 The procedure for document action is described in SOP QA- 011.

9.7.1.2 Master File.

9.7.1.2.1 The Master File contains all of the documents of the Pacific Agricultural Laboratory Quality System.

9.7.1.2.2 A Master File List of all documents is located at the front of the Master File. The Master File List shows the date of the most recent revision for each document.

9.7.1.2.3 Only current documents are maintained in the Master File. Each document contains a revision date in the lower right-hand corner of the document. This date corresponds to the date of origin of the document, or the date of the most recent revision to the document. Any document undergoing revision is removed from the Master File and the approved, revised document is then re-entered into to the Master File. Documents removed from the Master File are maintained for historical purposes as described in 9.7.1.3.

9.7.1.3 Document Action Form

9.7.1.3.1 Each time any document is revised, an existing document is removed, or a new document is added to the Master File, a Document Action Form is completed that describes the specific nature of the document action taken.

9.7.1.3.2 If the action taken is document revision, the revised document is issued a new revision date, which appears on the lower right-hand corner of the document. The updated revision date is then entered into the Master

17 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory File List, replacing the earlier date. The obsolete document and the revised document are married to the Document Action Form and the three documents are entered into the Document Action File. Any additional copies of the obsolete document are destroyed.

9.7.1.3.3 If the action taken is document removal, the removed document is married to the Document Action Form and the two documents are entered into the Document Action File. The document title is removed from the Master File List. Any additional copies of the removed document are destroyed.

9.7.1.3.4 If the action is document addition, the new document is married to the Document Action Form and the two documents are entered into the Document Action File. The new document is then added to the Master File and the title (and document number, where appropriate) are added to the Master File List.

9.7.1.3.5 All Document Action Forms are filed chronologically.

9.7.2 All document actions must be approved by either the Laboratory Director or the Quality Manager.

10.0 SYSTEM AND PERFORMANCE AUDITS

10.1 Formal Quality System audits are performed at a minimum of once annually. A written report is prepared by the Quality Manager that describes the findings of the audit. The audit consists of an evaluation of laboratory performance, QA problems and their respective corrective actions, customer complaints, laboratory audits, and training. Reports also outline changes to the Quality System, status of SOPs, status of the Chemical Hygiene Plan, significant QA problems and accomplishments, safety issues, and certification status. Audit reports are submitted to a management team for review (see 10.6) and are maintained in the Quality System audit file.

10.2 Internal audits are performed at the discretion of the Quality Manager.

10.3 The procedure for performance of annual internal audits is described in SOP QA- 006.

10.4 A formal annual management review of audit reports occurs one month following the completion and submission of the internal audit report. The review consists of the Laboratory Director, the Laboratory Manager, and the Quality Manager.

18 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory Based on the findings of the review, if necessary corrective actions are identified and action plans (time and events) are determined. Any corrective actions stemming from the internal audit review are documented on corrective action forms as per SOP QA-004.

10.7 Annual internal audits commence on February 1st, or as soon as possible thereafter.

10.8 Audit reviews commence no later than one month following completion of the laboratory audit.

11.0 DATA QUALITY OBJECTIVES

11.1 Pacific Agricultural Laboratory is committed to providing customers with the highest quality data obtainable that is accurate, complete, and legally defensible. To achieve this goal, Pacific Agricultural Laboratory has adopted the following practices.

11.1.1 Laboratory SOPs describe proper methods of sample handling and preservation, tracking, analysis, and reporting. All SOPs are readily available to all laboratory personnel and are updated on a regular basis. Any deviations from SOPs must be approved by management and be fully documented.

11.1.2 Analyses are performed by qualified personnel with appropriate training and supervision. Pacific Agricultural Laboratory encourages professional development and cross-training for its personnel.

11.1.3 Routine calibration and maintenance are performed on laboratory instrumentation to ensure correct and reliable operation.

11.1.4 Specific acceptance limits for precision are identified for each analytical procedure.

11.1.5 All phases of sample handling, analysis and data management are properly documented.

11.2 Data quality is routinely assessed to determine precision, comparability, and completeness.

11.2.1 Precision

11.2.1.1 Precision is a measurement of the extent of agreement among individual measurements made on a single sample under similar

19 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory conditions. Precision is determined by the use of triplicates and is expressed as percent coefficient of variance (% c.v.)

% c.v. = standard deviation/mean x 100

11.2.1.2 Acceptable precision limits are defined in method specific SOPs. Limits are based on published method protocols or are determined from in-house validation studies.

11.2.2 Comparability

11.2.2.1 Comparability is the expression of confidence with which one set of data may be compared with another set resulting from the measurement of the same property.

11.2.2.2 Comparability of data produced at Pacific Agricultural Laboratory is maintained by conforming to approved SOPs which define acceptable analytical procedures, control limits, and the units in which results are reported.

11.2.2.3 Pacific Agricultural Laboratory uses internal control performance samples to assess comparability.

11.2.3 Completeness

11.2.3.1 Completeness is a comparison of the amount of valid data actually obtained relative to the amount that was expected to be obtained.

11.2.3.2 Because Pacific Agricultural Laboratory is not involved in the sampling process, laboratory control over completeness is limited to ensuring that all data reported is accurate, precise, and properly documented.

11.2.3.3 If the amount of sample provided by the customer limits analysis or data reliability, the customer will be notified to allow for the possibility of resampling.

12.0 CHEMICALS, REAGENTS AND ANALYTICAL STANDARDS

12.1 All chemicals, reagents and analytical standards are of the highest quality obtainable in accordance with the specifications of the individual analytical method.

20 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 12.2 All chemicals, reagents and analytical standards are dated and initialed at the time that they are received and again when they are opened or prepared.

12.3 All chemicals and reagents are stored properly and discarded when shelf life has expired.

12.4 All prepared analytical standards and reagents are properly labeled and dated.

12.5 Wherever possible, standards are traceable. Certificates of analysis are retained in the Certificates of Analysis Binder.

13.0 EQUIPMENT

13.1 The laboratory is furnished with all of the equipment and instrumentation necessary to perform analysis. In the event that equipment outside the range of laboratory equipment is required, the relevant requirements of the equipment will be confirmed prior to use.

13.2 Major laboratory instrumentation is listed in Appendix A.

13.3 Where appropriate, all equipment and reference materials are labeled. Equipment calibration occurs either on an as-used basis (i.e., pH meter) or on a fixed schedule (i.e., drying oven). The calibration status of each piece of equipment on a fixed calibration schedule is documented on the individual equipment maintenance sheet.

13.4 Only laboratory supplies (glassware, etc.) that are of appropriate quality for the respective methods are used. Supplies are not placed into service until properly inspected to confirm that they meet manufacturer’s quality claims. A laboratory file maintains records from all suppliers of outside services and supplies. Procedures describing the purchase, receipt, and storage of supplies are described in SOP QA-007 and SOP QA-008.

13.5 Each piece of equipment has a dedicated file within the Equipment Master File containing equipment data sheets, manufacturer’s manuals and maintenance recommendations. Maintenance frequencies, maintenance history and calibration are located in the equipment log for each piece of equipment on equipment maintenance sheets.

13.6 Where appropriate, individual pieces of equipment have instrument specific calibration SOPs. Each piece of equipment is calibrated and/or verified before being placed into service.

13.7 Where appropriate, individual pieces of equipment have reference measurement standards used during calibration. These include:

21 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory

13.7.1 Drying Ovens-thermocouple thermometers calibrated to a NIST tracable thermometer.

13.7.2 Refrigerators and freezers-thermometers calibrated to a NIST tracable thermometer.

13.7.3 Mettler Toploader Balance–calibrated to a set of Class 1 NIST tracable calibration weights.

13.7.4 Sartorius Analytical Balance–calibrated to a set of Class 1 NIST tracable calibration weights including a 20 mg weight.

13.7.5 pH Meter-calibrated to a minimum of two pH solutions bracketing the pH range of the samples or reagents being measured.

13.7.6 Chromatography Intrument Systems-applicable analytical standard material(s) specific to an analytical method.

13.8 Calibration frequencies are as follows:

13.8.1 NIST tracable reference thermometer–annually

13.8.2 Class 1 reference weights-annually

13.8.3 Drying oven thermometers–twice annually

13.8.4 Refrigerator and freezer thermometers–twice annually

13.8.5 Mettler Toploader Balance–daily

13.8.6 Sartorius Analytical Balance–daily

13.8.7 pH meter - with every use

13.8.8 Chromatography Instrument Systems-as applicable for the specific analytical method

13.8.10 Records and documentation for each reference thermometer, reference weights and analytical standard materials are maintained in files.

13.8.11 Each piece of equipment has a specific SOP describing proper operation.

13.8.12 Each piece of equipment has a specific SOP describing proper maintenance.

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14.0 SAMPLE RECEIPT AND LOG-IN PROCEDURES

14.1 The proper sample log-in procedure is described in detail in SOP QA-002 which is used in conjunction with the Sample Log-in Form.

14.2 Samples are unpacked, inspected, and assigned a unique Pacific Agricultural Laboratory identification number. An identification number is assigned to each sample, which is entered onto the Sample Log-in Form.

14.3 A sample may be rejected due to a broken or leaking container, improper documentation, inadequate sample volume, etc. The customer will be notified of sample rejections and resampling recommendations will be provided to the customer. If samples are rejected, the reason for rejection is documented on the Sample Log-in Form.

14.4 Samples determined to be suitable for analysis are moved to designated storage areas.

14.5 Specific instructions for sample handling and preservation are detailed in method specific SOPs.

15.0 ANALYTICAL PROCEDURES

15.1 Pacific Agricultural Laboratory procedures are described in detail in Material Testing SOPs.

15.2 Whenever possible, analytical procedures are based on published, approved methods (e.g., American Society for Testing and Materials, etc.) and manufacturer’s recommendations. These procedures are referenced in method- specific SOPs.

15.3 The use of nonstandard methods or variations of standard methods will be validated and documented in laboratory notebooks prior to their use in the testing of customer samples. The authorization of both the Technical Manager and the Quality Manager must be obtained before nonstandard methods are implemented. Any method deviations are noted on the Materials Analysis Report Form.

15.4 Methods are selected based on industry requirements, the type of material to be analyzed, and the level of data quality expected by the Pacific Agricultural Laboratory customer base.

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15.5 All analytical procedures are tested to verify and document method ruggedness and precision as a means of determining method specifications for acceptance/rejection of sample test results.

15.6 Procedures must be approved by either the Laboratory Director or the Laboratory Manager before being implemented as routine laboratory methods.

15.7 Any deviation from methods during the analysis of a sample(s) will be noted on the final analytical report provided to the customer.

16.0 SAMPLE RETENTION AND DISPOSAL

16.1 Samples are retained at a minimum of 90 days from date of issuance of the final analytical report to the customer. Special arrangements may be made with the customer for the expedited return of samples or for extended retention.

16.2. All samples and laboratory wastes are disposed of in compliance with local, state, and federal regulations.

16.2.1 Nonhazardous solid materials (sand, silt, and clay) are disposed of in a sanitary landfill.

16.2.2 Nonhazardous liquid wastes are disposed of via sanitary sewer.

16.2.3 Hazardous materials are disposed of in accordance with all local, state, and federal regulations. Hazardous wastes are currently delivered to the Metro Hazardous Waster Center located in Portland, Oregon for disposal.

17.0 COMPUTERS

17.1 Pacific Agricultural Laboratory uses PC based computer systems throughout the laboratory.

17.2 All computer software is documented and suitable for use in documentation, data analysis, reporting, and the storage and retrieval of test data.

17.3 Computer software user manuals and support documentation are maintained on site.

17.4 The computer operating environment is routinely monitored to assure that it is suitable for proper computer operation and maintenance.

24 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 17.5 Computer access is restricted to authorized Pacific Agricultural Laboratory personnel. Each authorized individual must use a unique password to access Pacific Agricultural Laboratory databases.

17.6 Security of all Pacific Agricultural Laboratory computer files including documents, data, report forms, and communications is maintained with a computer back-up system as defined in SOP QA-005.

18.0 LABORATORY PERSONNEL TRAINING

18.1 Laboratory training is performed according to SOP QA-010 and consists of the following:

18.1.1 The trainee reads the Chemical Hygiene Plan.

18.1.2 The trainee reads the SOP that describes the specific method.

18.1.3 The procedure is performed by the trainee under the supervision of an experienced analyst.

18.1.4 The trainee must demonstrate method proficiency to the Laboratory Manager and proper understanding of method QA, safety practice, and documentation procedures.

18.1.5 Where appropriate, the trainee will perform proficiency testing on blinded samples that have a known composition.

18.1.6 Training is documented on an Employee Training Record and filed in the Pacific Agricultural Laboratory Training Log. Training requirements apply to both regular and temporary employees.

18.2 The Laboratory Manager is responsible for ensuring that only qualified analysts with proper recency of experience with the method(s) in use perform quality related functions.

18.3 Training records describing the date, nature of the training, and the outcome of the training are maintained by Pacific Agricultural Laboratory for each employee.

19.0 SAFETY

19.1 Pacific Agricultural Laboratory maintains a laboratory safety program that strives to remain in compliance with Occupational Safety and Health Association (OSHA) regulations.

25 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 19.2 Formal safety inspections are conducted every six months and potential hazards are noted for corrective action.

19.3 Pacific Agricultural Laboratory maintains and follows a Chemical Hygiene Plan as per 29 CFR 1910.1450.

19.4 Training sessions are provided periodically for full time laboratory staff.

19.5 Material safety data sheets are stored in a central location and are readily accessible to all Pacific Agricultural Laboratory employees.

19.6 All accidents and/or spills are documented on the Accident/Spill Report Form as described in SOP QA-009 by the Laboratory Manager.

20.0 ENVIRONMENTAL POLICY

20.1 Pacific Agricultural Laboratory is committed to minimizing environmental impact and to maintaining compliance with federal, state, and local regulations governing the environment.

20.2 All employees are expected to abide by and comply with all applicable environmental laws.

20.3 Pacific Agricultural Laboratory will meet or exceed environmental standards and regulations by requiring strict adherence to environmental laws, regulations, and standards by all employees and contractors.

20.4 Pacific Agricultural Laboratory will also practice the following:

20.4.1 Avoid unnecessary generation of wastes.

20.4.2 Promote recycling efforts

20.4.3 Render residual by-products environmentally compatible

20.4.4 Identify and respond to emerging environmental issues and concerns.

21.0 SUBCONTRACTING OF SERVICES

21.1 Pacific Agricultural Laboratory occasionally sub-contracts analyses that are not performed in-house. Pacific Agricultural Laboratory recognizes its responsibility for the quality of data generated and reported by sub-contractor laboratories and may require the following from sub-contractors.

26 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory 21.1.1 Sub-contractor laboratory QA Plan.

21.1.2 Sub-contractor laboratory inspections performed by a Pacific Agricultural Laboratory auditor.

21.1.3 Recent performance evaluations, including proficiency testing results.

21.1.4 Batch QA results.

21.1.5 Verifiable limits of detection

21.2 Additional, situation-specific information may be required based on specialized work subcontracted.

21.3 Pacific Agricultural Laboratory will maintain detailed records of its investigation into the competence of a sub-contractor and maintain a file containing information on each sub-contractor investigated.

21.4 Pacific Agricultural Laboratory will inform its clients of any work that is subcontracted and upon request, provide the customer with appropriate sub- contractor QA documentation.

27 Revised 7/10/06 Quality System Manual Pacific Agricultural Laboratory Appendix A, Analytical Instrumentation List

1. Agilent Technologies mass spectrometer. System includes an Agilent Technologies 6890 Plus gas chromatograph, Hewlett Packard 7683 autosampler, Hewlett Packard 5973 mass selective detector and turbomolecular vacuum pump. The GC-MS is controlled by a Hewlett Packard Enviroquant software system.

2. Hewlett Packard 6890 electron capture gas chromatograph. System includes a Hewlett Packard 6890 Plus gas chromatograph, Hewlett Packard 7683 autosampler, dual split/splitless inlets with EPC and dual electron capture detectors (micro-ECDs). The GC- ECD is controlled by a Hewlett Packard Chemstation software system.

3. Hewlett Packard 5890 FPD/FID gas chromatograph. System includes a 5890 Series II gas chromatograph equipped with autosampler, dual split/splitless inlets with EPC, flame photometric detector (FPD) and flame ionization detector (FID). The GC is controlled by a Hewlett Packard Chemstation software system.

4. Hewlett Packard 1050 high performance liquid chromatograph. System includes autosampler, quaternary pump, diode array UV detector, 1046A fluorescence detector and a Pickering post column reactor. The HPLC is controlled by a Hewlett Packard Chemstation software system.

5. Agilent 1100 high performance liquid chromatograph-mass spectrometer System includes autosampler, quaternary pump, diode array UV detector, and mass spectrometer. The HPLC is controlled by a Hewlett Packard Chemstation software system.

28 Revised 7/10/06 SOP-AM-033 Pacific Agricultural Laboratory Extraction of Imidazolinone Herbicides in Water

1.0 Scope and Application

1.1 This procedure describes the extraction of imidazolinone herbicides from aqueous samples. This method is applicable to all types of water including, but not limited to, drinking water, storm water, surface water, and groundwater.

2.0 Summary of Method

2.1 A 500mL aliquot of sample is acidified to pH 2 and 12.5g sodium chloride is added. Sample is shaken in a separatory funnel with three 50mL portions of dichloromethane. Organic layers are drained [through acidified sodium sulfate] into a round bottom flask, and concentrated by rotary evaporation (SOP-AM-027).

3.0 Interferences

3.1 Potential interferences may include contamination from glassware and solvents, and co-extracted materials from the sample matrix. Care must be taken to avoid and/or minimize these potential interferences.

4.0 Sample Handling and Preservation

4.1 Samples should be taken in 1-L amber glass bottles with a PTFE lined cap. 4.2 Samples are taken at neutral pH, and stored at 4°C prior to extraction. 4.3 All water samples shall be extracted within seven (7) days of sampling.

Revision Number: 1 1 Revision Date: 9/8/10 SOP-AM-033 Pacific Agricultural Laboratory 5.0 Apparatus and Instrumentation

5.1 1000 mL glass separatory funnel 5.2 500 mL graduated cylinder 5.3 600 mL beaker 5.4 250 mL round bottom flask 5.5 Large glass funnel 5.6 pH meter 5.7 Top-loading balance, accurate to ±0.01g 5.8 Magnetic stir bar 5.9 Magnetic stir plate 5.10 Rotary evaporator, Rotavap; Yamato RE50

6.0 Reagents and Supplies

6.1 Organic-free water, DI H2O 6.2 Methanol (MeOH) w/0.5% Formic Acid 6.3 Pesticide-grade Dichloromethane, DCM 6.4 6 N Hydrochloric Acid, HCl 6.5 Sodium chloride, ACS grade 6.6 Glass beads 6.7 [Glass wool]

6.8 [Acidified sodium sulfate, Na2SO4]

7.0 Procedure

7.1 For each sample, the necessary glassware items (separatory funnel, 600 mL beaker, and flat-bottom flask) are obtained, rinsed with Dichloromethane if necessary, and labeled with sample number. Beakers contain a magnetic stir bar, and two glass beads are added to each flat- bottom flask. Using a graduated cylinder, measure 500 mL of organic-

Revision Number: 1 2 Revision Date: 9/8/10 SOP-AM-033 Pacific Agricultural Laboratory free water for QC and transfer to a beaker with a stir bar. Likewise, measure and transfer 500 mL of sample into a beaker with a stir bar. 7.2 [Sodium sulfate funnels are prepared by placing a small plug of glass wool into a glass powder funnel, to which ~25g acidified Sodium Sulfate is added. Funnels are rinsed with ~10mL DCM, and solvent is drained into waste. A funnel is placed on each labeled collection flask.] 7.3 Using a 500 mL graduated cylinder, a 500 mL aliquot of sample is measured and transferred to the labeled 600 mL beaker. 7.4 Method Blank (BLK) consists of 500 mL deionized water in a 600mL beaker. This sample will be the negative control (QC) for the analysis. 7.5 Lab Control Sample/Lab Control Sample Duplicate (LCS/LCSD) each consist of 500 mL DI water in a 600mL beaker. Project specific spike compounds are added to each, and the standard log number and spike volume are recorded on extraction bench sheet. These samples will be the positive control (QC) for the analysis. 7.6 The pH of each sample and QC is adjusted to 2.0 by dropwise addition of 6N hydrochloric acid. 7.7 12.5 g of sodium chloride is added to each beaker, stirring until salt is completely dissolved. 7.8 The contents of each beaker are transferred into the appropriately labeled separatory funnel. Samples and QC are extracted by shaking three times with 50mL DCM. The lower (DCM) layers are drained [through the acidified sodium sulfate funnel] into the corresponding flat-bottom round flask.

7.9 [After all solvent is collected, Na2SO4 funnels are rinsed with ~20mL Dichloromethane, to optimize recovery of analytes.] 7.10 Extracts are concentrated to ~0.5 mL using rotary evaporation (SOP-AM- 027), and remaining solvent is evaporated to dryness under a steady stream of nitrogen gas.

Revision Number: 1 3 Revision Date: 9/8/10 SOP-AM-033 Pacific Agricultural Laboratory 7.11 Extract is transferred to labeled culture tubes as per SOP-AM-XXX (Rotavap) using MeOH w/0.5% Formic acid as final solvent. Final volume is 2mL for most Imidazolinone extractions. 7.12 Extracts should be stored in refrigerator until analysis.

8.0 Calculations

8.1 N/A

9.0 Quality Control

9.1 At a minimum, batch QC will include a method blank (MB), and a Laboratory Control Sample/Laboratory Control Sample Duplicate (LCS/LCSD). Additional QC will be performed if there are project and/or method specific requirements. An extraction batch consists of a batch of 20 consecutive samples extracted within 7 days. 9.2 Spike recoveries are calculated after analysis to evaluate extraction efficiency.

10.0 Reporting

10.1 N/A

11.0 References

11.1 American Cyanamid Method 2261 11.2 American Cyanamid Method M1900

Revision Number: 1 4 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

Imidazolinone Herbicides in Water by EPA 8321B

1.0 Scope and Application 1.1 This procedure is used to determine the concentrations of Imidazolinone herbicides in liquid matrices.

2.0 Summary of Method 2.1 A measured volume of sample is extracted using AM-033, Extraction of Imidazolinone Herbicides in Water. 2.2 Extracts are analyzed using liquid chromatography with mass spectroscopy (LC/MS) detection.

3.0 Interferences 3.1 Potential interferences may include contaminated solvents and extraction glassware, dirty chromatographic equipment, and co-extracted materials from the sample matrix. Care must be taken to avoid and/or minimize these interferences.

4.0 Sample Handling and Preservation 4.1 Store samples at 4oC out of direct sunlight. Water samples should be extracted within 7 days of sampling and analyzed within 40 days of extraction 4.2 Personal protection measures should be taken while handling solvents and samples.

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5.0 Apparatus and Instrumentation 5.1 Analytical balance, Sartorius model CP124S, accurate to 0.0001g. Calibration of balance shall be checked daily (SOP EQ-001). 5.2 N-EVAP evaporation manifold with heated water bath 5.3 HPLC System 5.3.1 Agilent 1100 HPLC system equipped with binary pump, autosampler, solvent degasser, and single quadrapole mass spectrometer. 5.3.2 Agilent Chemstation software 5.3.3 Analytical Column – C18 reverse phase column, 100mm x 3.0mm ID, 2.5 μm particle size, Agilent Zorbax SB-C18 or equivalent.

6.0 Reagents and Supplies 6.1 Organic-free reagent water 6.2 Methanol, Chemsolve, HPLC Grade 6.3 Acetonitrile (ACN), Chemsolve, HPLC Grade 6.4 Formic Acid, EMD, ACS Grade 6.5 Luer lock tipped syringe 6.6 Screw capped tubes with Teflon lined lids 6.7 13mm 45 μm nylon syringe filters 6.8 Auto sampler vials with PTFE lined caps 6.9 Volumetric flasks, class A 6.10 Gas tight syringes with PTFE tipped plungers 6.11 HPLC/MS Tuning Standard – Aglient ES Tuning Mix G2421A

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7.0 Procedures 7.1 Sample Extraction: 7.1.1 Extract waters via the procedure outlined in Pacific Agricultural Laboratory SOP AM-033 “Extraction of Imidazolinone Herbicides in Water”. 7.1.2 Store extracts in refrigerator until analysis. 7.2 Solvent exchange of water extracts: 7.2.1 Transfer a 1 ml aliquot of the sample extract to a culture tube. Mark the meniscus of the liquid in the tube. 7.2.2 Evaporate the solvent under a steady stream of nitrogen using the N-Evap evaporation manifold. 7.2.3 Reconstitute the extract as follows: add 500 uL methanol, then 500 uL Mobile Phase A (95% organic free water, 5% ACN, 0.05% formic acid). . 7.2.4 Filter the sample extract into an autosampler vial through a 45 μm 13 mm syringe filter using a luer tipped syringe. 7.2.5 Cap the vial and label with appropriate moniker. 7.3 Preparation of HPLC mobile phase: 7.3.1 The mobile phase is contained in two reservoirs, one containing the aqueous portion (Mobile Phase A) and one containing the organic( Mobile Phase B) portion. 7.3.2 Prepare Mobile Phase A by combining 950 mL of organic free water, 50 mL ACN, and 0.5 mL formic acid. 7.3.3 Prepare Mobile Phase B by combining 950 mL of ACN, 50 mL organic free water, and 0.5 mL formic acid. 7.4 Chromatographic conditions: 7.4.1 Flow rate: 0.40 mL/minute 7.4.2 Injection volume: 10 ul 7.4.3 Column Temperature: 45 oC

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7.4.4 Solvent Gradient: Time %A %B 0.0 80 20 1.5 80 20 8.0 30 70 10 30 70 7.4.5 Re-equilibration time: 3 minutes, 80% A/20% B

7.5 Mass Spectrometer Conditions: 7.5.1 Ionization Mode: API-Electrospray o 7.5.2 Drying Gas: N2, 11.0 L/min, 250 C 7.5.3 Nebulizer Pressure: 30 psig 7.5.4 Capillary Voltage: 1500 V 7.6 Mass Spectrometer Detector settings: 7.6.1 Settings for use in MS data acquisition (SIM ions and fragmentor voltages) vary by analyte and are displayed in Table 2 of the Appendix (12.2). 7.7 If the peak areas of the sample signals exceed the calibration range of the system, dilute the extract as necessary and reanalyze the diluted extract.

Revision Number: 1 4 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

7.8 Calibration: 7.8.1 Electrospray MS System: The MS system is calibrated for accurate mass assignment, sensitivity, and resolution using the Agilent ES Tuning Mix G2421A. The following masses are calibrated in positive and negative ionization modes: MASS POSITIVE NEGATIVE 1 118.09 112.99 2 322.05 431.98 3 622.03 601.98 4 922.01 1033.99 5 1521.97 1633.95

Tune parameters are adjusted to ensure ions are present at each of the masses with counts >50000 and peak widths within the range of 0.60 – 0.70 amu. 7.8.2 Stock Standards: Individual analyte stock standards are made at concentrations between 500-1000 Pg/ml by transferring 25-50 mg neat standard to a 50 mL class A volumetric flask, dissolving the neat standard in acetonitrile or methanol, and diluting to the mark with acetonitrile or methanol. Stock standards prepared from neat standards may be used for a maximum of two years. Alternatively, a solution containing 1000 Pg/ml of analyte may be obtained from ChemService or other reputable manufacturer and used as a stock standard. In this case, the stock standard may be used until the expiration date provided by the manufacturer. 7.9.3 Working Standards: A 10 Pg/ml working standard is made by transferring appropriate amounts, depending on initial concentrations, of stock standards to a 10 mL class A volumetric flask and diluting to the mark with methanol or acetonitrile. The

Revision Number: 1 5 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

amount of stock standard to transfer will range between 100-200 PL and is calculated using the formula: Amt. Stock Std.(PL) = [Final Conc. (10Pg/ml)] x [Final Vol. (10ml) Initial Stock Conc. (μg/μL) The working standard solution is transferred to an appropriately labeled screw cap tube and may be used for a maximum of one year. 7.9.4 Preparation of external standard calibration curve: an appropriate aliquot of the working standards are added to an autosampler vial and diluted to 1 ml with Mobile Phase A. A minimum of 5 standards are prepared at the following suggested levels: 0.005 ug/ml, 0.010 ug/ml, 0.020 ug/ml, 0.05 ug/ml, and 0.10 ug/ml. The calibration range can be adjusted to meet expected levels in the samples. The calibration standards are prepared as follows: Calibration Aliquot Concentration Volume of Final level volume of aliquot(s) buffer volume 100 ng/ml 100 Pl 1000 ng/ml 900 Pl 1.0 ml 50 ng/ml 50 Pl 1000 ng/ml 950 Pl 1.0 ml 20 ng/ml 200 Pl 100 ng/ml 800 Pl 1.0 ml 10 ng/ml 100 Pl 100 ng/ml 900 Pl 1.0 ml 5 ng/ml 50 Pl 100 ng/ml 950 Pl 1.0 ml

7.9.5 The system is calibrated prior to the injection of a set of sample extracts. After injecting a set of standards, a linear calibration curve is prepared. Exclude the origin as a point. The R value of the generated curve should be 0.99 of better. If the calibration fails to meet these criteria, the cause of the deviation should be rectified and the system recalibrated.

7.9.6 The calibration is verified by injecting a CCV at the mid point concentration of the calibration curve after no more than twenty Revision Number: 1 6 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

samples. If the response deviates by more than +/- 15% from the initial calibration, the system should be recalibrated and the samples bracketed by the either the initial calibration or the prior passing CCV and the failing CCV should be reanalyzed. If the CCV is >15% of initial calibration, the samples bracketed by the either the initial calibration or the prior passing CCV and the failing CCV can be used if the sample contains no detectable residues.

8.0 Calculations 8.1 Water Samples:

amount f/curve (ng/ml) x final volume (ml) x dilution factor = result (ug/liter, ppb) sample volume (ml)

9.0 Quality Control 9.1 Initial Demonstration of Proficiency – the laboratory shall demonstrate initial proficiency with each sample preparation technique, by generating data of acceptable accuracy and precision for target analytes in a clean matrix. The laboratory must also repeat the demonstration whenever new staff is trained or significant changes in instrumentation are made. 9.1.1 Calculate the average recovery and the standard deviation of the recoveries of the four QC reference samples. Refer to Section 8.0 of EPA Method 8000 for procedures in evaluating method performance. 9.2 Method Reporting Limits (MDLs) 9.2.1 The MDL is defined as the minimum concentration of a substance that can be measured and reported with 99% confidence that the analyte concentration is greater than zero and is determined from analysis of a sample in a given matrix type containing the analyte.

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9.2.2 The extraction and analysis of seven replicates of a spiked sample determine the MDL. 9.2.3 Multiply the standard deviation of the seven replicate results by the one sided 99%t-statistic (3.14) to obtain the MDL for each analyte. 9.2.4 These results are kept on file and should be re-evaluated annually, when significant changes in instrumentation are made, or when new staff are added. 9.3 Sample Quality Control for Preparation and Analysis 9.3.1 The laboratory will have procedures for documenting the effect of matrix on method performance. 9.3.2 Water matrix – minimum QC samples shall include a method blank (MB), Laboratory Control Sample (LCS), and a Laboratory Control Sample Duplicate (LCSD). A matrix spike may be prepared and analyzed provided there is adequate sample. 9.4 QC Frequency – an analytical batch is defined as a set of no more than 20 samples extracted within 14 days. The QC frequency for each analytical batch is as follows: Method blank – 5% Matrix Spike/Matrix Spike Duplicate – 5% Laboratory Control Sample/Laboratory Control Sample Duplicate – 5% 9.4.1 In house method performance criteria for spike and surrogate compounds should be developed using guidance found in Section 8.0 of EPA Method 8000. 9.4.2 If the recovery data is outside acceptance limits, the samples should be re-extracted and/or the data flagged as necessary.

Revision Number: 1 8 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

10.0 Reporting 10.1 If all QC criteria have been met, the data is then compiled and a report is generated, including sample raw analytical results and QC data, and submitted to the client.

11.0 References 11.1 EPA Method 8321B, SW-846 Revision 2, December 2007. 11.2 Pacific Agricultural Laboratory Quality System Manual. 11.3 EPA Method 8000B, SW-846 Revision 2, December 1996. 11.4 SW-846, Chapter One, Revision 1, 1992.

12 Figures and Appendices 12.1 Table 1 - Analyte list and reporting limits 12.2 Table 2 – Mass Spectrometer Data Acquisition Settings

Approved:

Date:

Revision Number: 1 9 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

TABLE 1 ANALYTE LIST AND LIMIT OF QUANTATITION (LOQ) Analyte LOQ, ug/L

Imazamox 0.02 Imazapic 0.02 Imazapyr 0.02 Imazethapyr 0.02

TABLE 2 – MASS SPECTROMETER DATA ACQUISITION SETTINGS

Time SIM Ions Fragmentor Voltage Capillary Voltage 0.00 220,222,234, 200 2000 V 248,262,277, 278,290,293, 306,307

Revision Number: 1 10 Revision Date: 9/8/10 SOP AM-034 Pacific Agricultural Laboratory

TABLE 3 – SIM IONS FOR IDENTIFICATION/QUANTIFICATION

Analyte Quantification Ion Qualifier Ions Ionization Fragmentor Mode Voltage Imazamox 306 307,278 positive 200 Imazapic 293 277,220 positive 200 Imazapyr 262 234,222 positive 200 Imazethapyr 290 262,248 positive 200

Revision Number: 1 11 Revision Date: 9/8/10

APPENDIX 6.

General Site Safety & Materials Handling Guidelines and Procedures for Spartina Control Projects in the San Francisco Estuary General Site Safety & Materials Handling Guidelines and Procedures for Spartina Control Projects in the San Francisco Estuary

IMPORTANT DISCLAIMER AFFECTING LEGAL RIGHTS. The San Francisco Estuary Invasive Spartina Project (ISP) compiled this document to provide a suggested set of general guidelines and procedures that are consistent with the mitigation measures required by the ISP EIR [spelled out](“PEIR”). These guidelines may be used by ISP partners and contractors as a minimal baseline, consistent with the PEIR, for planning and implementing site-specific Spartina control work in the San Francisco Estuary. These general guidelines are not intended to cover all safety procedures and precautions that may be necessary, nor are they intended to substitute for a comprehensive set of safety procedures and precautions for any particular control work or site. The ISP and the Conservancy make no warranties, assur- ances or representations of any kind with respect to the scope, extent, propriety or effectiveness of the suggested procedures that are described in these guidelines. Each ISP partner is responsible for and should independently develop and implement all appropriate safety precautions and procedures needed for the control work it undertakes. As a condition to use of these guidelines, each ISP partner and any of its contractors agree that the ISP and the State Coastal Conservancy (“Conservancy”) shall not be responsible for the acts or omis- sions of any ISP partner, or its contractors or volunteers, and agree to release and hold harmless the ISP and the Conservancy from any claims or liability, in connection with the development or implementation of site-specific safety procedures and precautions.

May 2005 Table of Contents

Table of Contents...... i

Emergency Information...... 1 Emergency Phone Numbers...... 1 Nearest Hospital...... 1

Sensitive Receptors...... 2 Listing of Sensitive Receptors ...... 3

Site Safety Protocols for Working in Marshes, Mudflats, and other Tidal areas of the San Francisco Estuary ...... 4 Teams...... 4 Channels...... 4 Mudflats ...... 5 Chemical or Physical Hazards ...... 5 Tides...... 5 Weather...... 6 Communication...... 6

Herbicide Handling, Spill Prevention and Spill Response ...... 7 Herbicide Use...... 7 Herbicide Storage...... 7 Container Disposal...... 7 Spill Response...... 8 Preventing Spills ...... 9

Spray Drift Reduction ...... 10

Petroleum Fuel Spill Prevention and Response...... 12

Herbicide Information...... 13

Invasive Spartina Control Plan 2005-7 i General Site Safety & Materials Attachment 4 Handling Guidelines Emergency Information

Emergency Phone Numbers In case of any emergency, call 9-1-1, and follow dispatcher instructions

Pesticide Emergency: o Call the ChemTrec (Chemical Transportation Emergency Center) emergency number, 1-800-424-9300, for instructions on how to handle any pesticide emergency o Emergency involving Monsanto products (Aquamaster): 314-694-4000 o Emergency involving Dow products (Rodeo): 800-369-2436 or 979-238-2112 o Emergency involving BASF products (Habitat): 800-832-4357

Chemical Spill into Marsh or other Waters: o Report all spills within 24 hours to the San Francisco Regional Water Quality Control Board: 510-622-2369

Nearest Hospital The following space is provided for the ISP grant recipient or its contractor(s) to provide the location and directions to the closest hospital.

Name: ______

Address: ______

______

Phone: ______

Directions to hospital: ______

______

A map should be attached or otherwise made available on site.

Invasive Spartina Control Plan 2005-7 1 General Site Safety & Materials Attachment 4 Handling Guidelines Sensitive Receptors When applying herbicides for non-native Spartina control, care must be taken to protect human health, and particularly “sensitive receptors” that may be located near the application area. Sensitive receptors include hospitals, schools, and residences near the bay margin that are in close proximity (e.g., within 0.25 mile) to areas being treated with herbicide. The potential presence of sensitive receptors must be evaluated on a site-specific basis. The Ad- jacent Land Uses section in the site-specific Invasive Spartina Control Plan contains some information regarding potential sensitive receptors at each sub-area. In general, sensitive receptors are most likely to occur at sites located in the Corte Madera Creek watershed in Marin County, and along the shorelines of Alameda, Hayward and San Leandro in Alameda County. Birders, bicyclists, joggers, pedestrians, and users of recreational facilities (including parks, marinas, launch ramps, fishing piers, and beaches) that sur- round the Bay also could be sensitive receptors. The ISP grant recipient(s) and their contractors are responsible for fully identifying and protecting sensitive receptors. To minimize risks to the public, mitigation measures for herbicide treatment methods must be implemented by entities engaging in treatment activities. Such measures include, but are not limited to, the following: 1. Post signs for sensitive receptors within 500 feet. At least one week prior to application, post signs informing the public of impending herbicide treatment at prominent locations within a 500-foot radius (approximately 1/10 mile) of treatment sites where homes, schools, hospitals, or businesses could be affected. Schools and hospitals within 500 feet of any treatment site should be separately noticed at least one week prior to the application. 2. Avoid aerial spraying near sensitive receptors. Do not conduct aerial spraying within 0.25 mile (1,320 feet) of a school, hospital, or other sensitive receptor location. 3. Minimize drift. Manage herbicide application to minimize potential for herbicide drift (See Spray Drift Reduction, page 11 of this document). Herbicide must not be applied when winds are in excess of 10 miles per hour or when inversion conditions exist, or when wind could carry spray drift into inhabited areas. 4. Post signs at access points. Post colored signs at and/or near any public trails, boat launches, or other potential points of site access 24 hours prior to treatment. The signs should inform the public that the area is to be sprayed with glyphosate and/or imazapyr herbicide for weed control, and that the spray is harmful if inhaled. The signs should advise “no entry” for humans and animals until eight (8) hours after treatment, and the treatment date and time should be stated. A 24-hour ISP contact number may be provided. 5. Avoid high use areas. Avoid application of herbicides near areas where the public is likely to contact water or vegetation. For example, avoid applying herbicide in or adjacent to high use areas within 24 hours of high use times, such as weekends or certain holidays. If a situation arises that makes it necessary to treat high-use areas during such times, the areas should be closed to the public before, during, and after treatment.

Invasive Spartina Control Plan 2005-7 2 General Site Safety & Materials Attachment 4 Handling Guidelines Listing of Sensitive Receptors The following spaces are provided for the ISP grantee and its contractors to list sensitive receptors within 0.25 mile of the herbicide treatment site. This information should be made available in advance to herbicide application contractors. Schools Hospitals Name

Address

Contact

Phone

Name

Address

Contact

Phone

Name

Address

Contact

Phone

Residences Name

Address

Contact

Phone

Name

Address

Contact

Phone

Name

Address

Contact

Phone

Invasive Spartina Control Plan 2005-7 3 General Site Safety & Materials Attachment 4 Handling Guidelines Site Safety Protocols for Working in Marshes, Mudflats, and other Tidal areas of the San Francisco Estuary Tidal lands of the San Francisco Estuary present many unique hazards to workers who must access intertidal areas during the performance of their activity. The following is a summary of some of the hazards one may encounter when accessing these areas, and suggested precautions. There is no assertion made here, either stated of implied, that this list is comprehensive of all hazards that could possibly be encountered while in intertidal areas of the Estuary. Caution should be exercised at all times while in these areas, and common-sense danger avoidance techniques should be employed.

Teams Always travel with a partner when entering or working in marshlands. There are many hidden hazards associated with marshland travel and work that are not readily discernable at first glance. A team of at least two individuals adds a necessary level of safety for any work or activity taking place in the marsh. Supervisors should provide daily worker safety briefings prior to commencing work on site. These briefings need not be exhaustive, but should include any new information the supervisor may have obtained about the work conditions on the site, weather conditions, team assignments, equipment condition, or other pertinent issues.

Channels Watch for hidden channels and holes in the marsh plain as you traverse the marsh. Often smaller channels in the marsh are obscured by vegetation. These channels can be quite deep, and may result in a sprain or pulled muscle, or possible fractures. Use a probe, like a stick or staff to check ahead of your path for unseen channels. Keep alert for vegetation changes, like Grindelia (Gumplant), which grows along channel edges and may indicate a hidden channel. Channel banks can sometimes be quite soft, and the mud that lines the channel can often be unstable. It is not unusual to sink deeply into these muds. This could be dangerous during an incoming tide. Always probe the mud within channels to test its ability to support your weight before stepping forward. Channels often block direct routes through the marsh. These channels can be quite small or very large. Great caution should be observed when considering crossing these channels. It may seem worthwhile in most cases to leap across the channel to get to the other side. This should only be done on the smaller channels, when your partner is able to follow, when you have surveyed the route for alternate paths around the channel, and at low tide. Large channels should be avoided entirely, and placing driftwood bridges over these channels is not advised. Workers may fall from unstable bridges into larger channels and risk injury, drowning, hypothermia, or equipment destruction. It is best in these situations to find a way around the channel.

Invasive Spartina Control Plan 2005-7 4 General Site Safety & Materials Attachment 4 Handling Guidelines Mudflats Mudflats at low tide can be quite dangerous to the unprepared. Often these flats are extremely soft, making travel over them slow and messy. Without proper footwear made for travel over mudflats (called “mudders”), workers may sink up to their thighs in mud. If stuck deeply in a mudflat or channel bottom, you can often extract yourself by spreading out your weight over the mud by, in effect, laying or crawling on the muddy surface. Twisting your ankles within your boots or waders also works to extract your feet from the mud. Assistance from your partner in the marsh can be essential during these situations. If feet sink into soft mud or quicksand, do not make violent movements in an attempt to get free. If boots or waders become stuck, slip one foot out gradually, rest the leg on the surface and gradually free the other leg. Lying on the surface and spreading the weight can avoid sinking. Move to firm ground using a “leopard crawl” (spread eagled, face down, keeping the maximum area of the body in contact with the ground at all times).

Chemical or Physical Hazards Many marshland areas have been historic sites of dumping or disposal. Many marshes have accumulated debris or wrack that contains all manner of refuse. As a result, some areas have large amounts of this waste material, and in some cases, toxic waste or hazardous chemicals. Supervisors should be made aware of any known chemical or toxic waste issues associated with a site and take appropriate precautions. Workers should be notified prior to the beginning of operations within the marsh what the condition of the marsh is relative to toxic or hazardous substances, and be appropriately equipped. Wear footwear capable of resisting puncture by sharp objects. Nails, glass, chunks of concrete, rusty metal and other debris can severely injure workers without appropriate footwear. Ideally, workers should wear hip or chest waders with reinforced soles, that are resistant to puncture, tearing or chemicals. In areas where there are known concentrations of toxic or hazardous substances, a site- specific safety plan should be prepared and an appropriately trained hazardous materials expert should supervise work. At a minimum, workers should wear protective gloves and eyewear, long-sleeve shirts, and thoroughly wash all clothing subsequent to work in the marsh. Workers should also thoroughly wash themselves with soap and water following work activities. If potentially toxic or hazardous materials are discovered during work activities, the area should be marked and reported to the appropriate authorities (the County hazardous Ma- terials Office and/or the Regional Water Quality Control Board). The area should be avoided until the material has been assessed and/or removed from the site.

Tides All workers in the marsh shall be made aware of the tidal schedule prior to work in the marsh. Work shall commence on an ebb tide and cease on the incoming tide or earlier. Allow ample time to return to non-tidal areas before the incoming tide starts to advance across the work site. This general rule may be modified in higher marsh habitats where

Invasive Spartina Control Plan 2005-7 5 General Site Safety & Materials Attachment 4 Handling Guidelines tidal action is lessened, but workers should always be alert and mindful of working in situations where the incoming tides may trap them, and allow ample time for exiting the marsh prior to an incoming tide. If in doubt, get out. Tides can rise extremely quickly in some areas, and it is possible that rising tides may outpace the ability of workers to out- run the increasing water levels, especially in soft muds or heavily vegetated marsh plains.

Weather It is always important to monitor weather conditions prior to and during work activities within the marsh. Wind, rain, fog or other inclement weather can mean the difference between a safe work site and an extremely dangerous one. Winds usually occur in the early afternoon or late morning during the summer months, though dangerous weather patterns can occur at any time of the year. Rainfall may subject workers to hypothermia if unprepared, or may result in potentially dangerous floodwaters. Winds can increase wave action, whip up salt spray or dust. Fog can decrease workers ability to communicate or dis- cern potential hazards in the marsh. It is ill advised to go into marshland terrain in bad visibility. For all work performed in the intertidal areas of the Estuary, workers or supervisors should check weather forecasts prior to commencing work on the site, should monitor weather conditions for any changes while on site and should modify work plans accordingly to insure the safety of all personnel.

Communication Open lines of communication between workers in the marsh must be maintained. When more than one team will be working in the marsh at any one time, it is advisable to have a communication link to a land base and between individual teams for safety. In the case of injury, discovery of hazardous materials, endangered species, or cultural artifacts, or for other reasons, land-based assistance can be contacted from the field for immediate help or first aid. On the ground coordination via phone or walkie-talkie when crews are spread out over the marsh can help to avoid dangerous situations.

Invasive Spartina Control Plan 2005-7 6 General Site Safety & Materials Attachment 4 Handling Guidelines Herbicide Handling, Spill Prevention and Spill Response The following information and practices are to be incorporated into herbicide-based Spartina control operations associated with the San Francisco Estuary Invasive Spartina Project (ISP).

Herbicide Use All herbicides shall be applied by or under the direct supervision of trained, certified or licensed applicators and in accordance with the product label On-site mixing and filling operations shall be confined to areas appropriately bermed or otherwise protected to minimize spread or dispersion of spilled herbicide or surfactant into surface waters

Herbicide Storage Proper herbicide storage is one of the keys to using herbicides safely. Always wear rub- ber gloves when handling herbicides in storage, and review product labels for specific storage instructions. General rules for herbicide storage include: Keep all herbicides in their original containers. Store herbicides in a locked shelter away from children and animals. Store herbicides in a dry cool and well- ventilated area. DO NOT subject herbicides to freezing or extremely high temperatures. Store herbicides separately from seed, fertilizer, insecticides and food. Make periodic inspections of storage facilities and storage containers. Check for possible leaks, spills and other similar problems. Keep appropriate absorbent material in the storage area at all times as well as a plastic container for storing damaged material. Reject any broken or leaking containers when herbicides are delivered. Do not store herbicides in office or break areas where employees congregate.

Container Disposal Empty herbicide containers must be disposed of according to government regulations or be returned to the manufacturer for disposal. Empty containers not returned to the manufacturer can be handled according to the procedures below, as long as local, state and federal laws are followed: Triple rinse containers with water. Always pour the rinse-water into an appropriate receptacle. Rinsed containers should be disposed of in a landfill approved for pesticide disposal or in accordance with applicable government procedures. Check with your

Invasive Spartina Control Plan 2005-7 7 General Site Safety & Materials Attachment 4 Handling Guidelines supervisor to find out if and when herbicide containers may be handled in this manner.

Spill Response Under all circumstances, it is the responsibility of the applicator to assure that all precautions are taken prior to initiating work to assure protection of water quality and the environment. The applicator is also responsible for the provision of a Spill Response Kit that is appropriate for the work being undertaken.

The following procedures should be followed in the case of a non-petroleum chemical spill: Put on protective gloves, eyewear, a long-sleeved shirt and pants before cleanup If a container is leaking, immediately transfer the remaining herbicide to another appropriate container to prevent further spillage If the herbicide was spilled on a person, remove the contaminated clothing and rinse the product from the body. If necessary, perform appropriate first aid. Cover the spill area with an absorbent material to soak up the herbicide. Common cat litter, sawdust, soil or sand can all be used for this purpose. Consult the manufacturer for more specific clean up recommendations. Remove any contaminated items from the spill area to prevent further contamination Remove the absorbent material with a broom and or shovel after the spill has been absorbed. Make sure all contaminated soil is removed from the spill area as well. Place the contaminated soil and absorbent material into a suitable container, and dispose of the container in an approved landfill area Do not wash down the area with water using a high pressure hose. You may spread the spill and make the herbicide more difficult to contain and clean up. When a spill occurs on a site, or is large enough that you need help to contain of clean it up, contact a supervisor immediately. In case of a major spill, call the manufacturer or ChemTrec (Chemical Transportation Emergency Center), 1-800- 424-9300.

Spill Response Kit A Spill Response Kit should provided at the work site and be immediately accessible to all personnel. Some or all of the following items may be included in a Spill Response Kit. Consider site-specific conditions and the chemicals to be used to determine which of the following items are appropriate. PVC Gloves or equivalent (to mid forearm) Half-face respirator equipped with approved pesticide cartridge PVC boots or equivalent Chemical resistant splash goggles

Invasive Spartina Control Plan 2005-7 8 General Site Safety & Materials Attachment 4 Handling Guidelines Spray Drift Reduction

Definition of The Department of Pesticide Regulation (DPR) defines pesticide drift pesticide drift as the pesticide that moves through the air and is not deposited on the target area at the time of application. Drift does NOT include movement of pesticide and associated degradation compounds off the target area after application (e.g., translocation, volatilization, evaporation, or the movement of pesticide dusts or pesticide residues on soil particles that are windblown after application.)

The pesticide Pesticide drift, particularly from agricultural fields, has been known to drift issue impact adjacent residential areas, cause damage to non-target crops, and contaminate the environment.

How does Low levels of pesticide drift may occur from all types of pesticide ap- pesticide drift plications. Pesticide drift becomes unacceptable when pesticides are occur? applied by imprecise methods or under environmental conditions that prohibit the applicator from maintaining control over the path the pesticide takes once it leaves the application equipment.

The San Francisco Estuary Invasive Spartina Project (ISP) has identified the use of herbicide as a critical component of its Spartina Control Program. The herbicide used for Spartina Control is glyphosate (Rodeo or Aquamaster), a product with exceptionally low toxicity, approved by U.S. EPA and the State of California for use in sensitive aquatic and estuarine environments. The human health risks associated with glyphosate are very low, and it requires no special personal protection measures for handling and application. One of the surfactants approved for use on ISP projects, LI-700, is corrosive and can cause eye, skin, and respiratory irritation in humans. In any case, it is desirable to minimize exposure of humans or non-target plants to pesticide drift. The ISP requires that all herbicide application under the Control Program be managed to minimize spray drift to protect human health and the environment. Application of herbicide and surfactants in accordance with product labels (including the Supplemental La- beling for Aerial Application in California) will minimize spray drift. In addition, the ISP recommends the following: 1. For aerial application of herbicide mixtures: a. Application should be by helicopter; no airplane application should be used. b. Nozzle orifices of broadcast sprayers should be directed backward. c. Flow of liquid from each nozzle should be controlled by a positive shutoff system. d. Spray nozzles should be adjustable to allow control of droplet size. Use up to 1500 microns for windy conditions. e. Boom pressure should not exceed the manufacturer’s recommended pressure for the nozzles being used. f. Herbicide should be applied only when wind speed is three to 10 miles per hour at the application site, as measured by an anemometer positioned four feet above the ground.

Invasive Spartina Control Plan 2005-7 10 General Site Safety & Materials Attachment 4 Handling Guidelines g. Discharge should start only after entering the target site; discharge height should not exceed 10-15 feet above the crop or target; discharge should be shut off whenever necessary to raise the equipment over obstacles; discharge should be shut off before exiting the target site. 2. For ground application of herbicide mixture by vehicle-mounted or towed ground equipment: a. Herbicide should be applied only when wind speed is 10 miles per hour or less at the application site, as measured by an anemometer positioned four feet above the ground. b. Discharge should start after entering the target site; discharge should be shut off before exiting the target site.

Invasive Spartina Control Plan 2005-7 11 General Site Safety & Materials Attachment 4 Handling Guidelines Petroleum Fuel Spill Prevention and Response Spills of gasoline or other petroleum products, required for operation of motorized equipment, into or near open water could degrade water quality, with potential for toxicity of contaminant bioaccumulation. Several types of equipment used for treatment of Spartina may present opportunities for petroleum spills. Equipment used in Spartina control activities include: Amphibious tracked vehicles Spray trucks Water-based excavators (e.g. Aquamog) Gas-powered mowers (e.g. Weed-Whackers) Air boats and outboard motor boats Fueling Fueling of amphibious tracked vehicles, spray trucks or land-based excavators should be done offsite at fueling stations or suitable staging areas. A suitable staging area shall be equipped with sufficient protection to prohibit a petroleum spill from migrating beyond the immediate fueling area (e.g., an impermeable plastic tarp set between raised berms, a catch basin or similar portable device). Water-based excavators, airboats and outboard motor boats shall be fueled offsite at commercial fueling stations or designated locations such as equipment maintenance yards. When fueling is done on or adjacent to treatment sites, a spill prevention and response plan must be prepared and implemented. A copy of this plan shall be provided to the Invasive Spartina Project at [email protected]. Gas powered, hand held machinery (e.g., weed whackers) shall be refueled on a non- absorbent tarp or mat placed under machinery to catch any spills. In addition to spills during refueling operations, small amounts of oil or fuel may leak from improperly maintained equipment. Before using any equipment in the marsh, check to make sure that it is in good working order with no signs of leakage or corrosion that might indicate the potential for inadvertent spills on the work site. Transport vessels and vehicles, and other equipment (e.g., mower, pumps, etc.) shall not be serviced or fueled in the field except under emergency conditions. Under all circumstances, it is the responsibility of the applicator to assure that all precautions are taken prior to initiating work to assure protection of water quality and the environment. The applicator is also responsible for the provision of a Spill Response Kit that is appropriate for the work being undertaken.

Invasive Spartina Control Plan 2005-7 12 General Site Safety & Materials Attachment 4 Handling Guidelines Vice grip pliers Phillips head screwdriver (2) Shovels Brooms, dustpan Clay granules or a sawdust Activated charcoal or other appropriate absorbent material First aid kit Tyvek coveralls (2 pair) or neoprene coveralls Recovery drums DOT triangular reflector kit Source of clean water and soap In the case of refueling or mixing activities planned on open mudflats the spill response kit should include a portable wet vacuum or other pumping equipment

Preventing Spills The following procedures will help to minimize the risk of spills occurring: Keep bags and cardboard containers dry at all times Prevent or correct leaks in herbicide containers and application equipment Properly dispose of all empty pesticide containers Tie down or otherwise secure containers when transporting pesticides to prevent them from falling from a vehicle Store herbicides only in their original containers or properly labeled service containers Stay alert and attentive when handling or using herbicides Where on-site or in-field transfer of liquid chemicals (herbicide mixtures, fueling operations) are planned,

Invasive Spartina Control Plan 2005-7 9 General Site Safety & Materials Attachment 4 Handling Guidelines Herbicide Information This section provides product labels and Material Safety Data Sheets (MSDS) for herbicides and adjuvants that have been evaluated and approved for use in controlling non- native Spartina in the San Francisco Estuary. Product labels and MSDSs contain important information to help protect human health and the environment, and they should be included as a part your Site Safety Plan. Included in this section are the following products:

Aquatic Herbicides: 1. Aquamaster (glyphosate-based herbicide) – Product label, California supplemental label for aerial application, and MSDS. 2. Habitat (imazapyr-based herbicide) – Temporary product label (pending Cali- fornia registration) and MSDS. 3. Rodeo (glyphosate-based herbicide) – Product label and MSDS.

Surfactants: 1. Agridex (non-ionic crop oil) – Product label and MSDS. 2. Competitor (methylated seed oil) – Product label and MSDS. 3. Cygnet Plus (non-ionic surfactant) – Product label and MSDS. 4. DyneAmic (silicone-based surfactant) – Product label and MSDS. 5. Kinetic (silicone-based surfactant) – Product Label and MSDS. 6. LI 700 (non-ionic surfactant) – Product label and MSDS. 7. Liberate (non-ionic surfactant) - Product label and MSDS.

Colorants: 8. Blazon (spray pattern indicator) - Product label and MSDS.

Please note that ONLY the aquatic herbicides and surfactants that are listed here are approved for use in Spartina control in the San Francisco Estuary. There are, however, other adjuvants (excluding surfactants), such as drift retardants, anti-foaming agents, and acidifiers, that may be used, provided the ISP Partner has reviewed the product information and found the product to pose no significant risk to human health or the estuarine environment. It is the responsibility of the applicator to obtain product labels and MSDSs for any products not included in this document. It is the responsibility of the applicator to assure that the most current product labels are obtained and followed.

Invasive Spartina Control Plan 2005-7 13 General Site Safety & Materials Attachment 4 Handling Guidelines