Note to the Trainer:

1. Note to the Trainer: • These slides are provided to standardize crews to collect ozone indicator data. The first 50 slides are introductory. You may decide not to use all of these slides, but the following concepts must be emphasized. • Provide visual evidence that ambient ozone concentrations are high across your sampling area. It is best to locate a current, regional map from the EPA web site, but if this is not possible, use the national map provided in slide #14. http://www.epa.gov/air/data/index.html • Clarify the Training Objectives. Review or adapt slides #4 and #5 to your session. • Clarify the distinction between the beneficial ozone layer and ground level ozone pollution. • Provide some indication as to why the ozone indicator is important to FIA and to the country as in slides #36, #37, #38. See slides #16, #17, #18, #19 as well. • Provide some evidence of state-level results from your own region if possible. • Do not abbreviate the procedural slides included here and numbered slides #52 - #105. • The symptom review slides are in a separate file. Review the species that are used in your region and focus on the best slides. There is a third file that provides 16 ozone injury slides for testing the trainees and/or for additional review and discussion. The slides will load faster if they are first copied from the CD to your hard drive. You should then be able to edit them as you see fit.

2. Air Quality Guide (ppb O3) Green = Good Yellow = Moderate Orange = Unhealthy for Sensitive Groups Red = Unhealthy Purple = Very Unhealthy

3. Eastern biosite Western biosite National Ozone Sampling Grid

4. TRAINING OBJECTIVES 1. With the Field Guide as a reference, the trainee demonstrates a working knowledge of the ozone biomonitoring procedures. 2. Given a complete suite of data from a real biosite, the trainee accurately enters the data using the electronic data entry program or data sheets. 3. Given actual plants or high quality images, the trainee correctly identifies ozone injury symptoms on the bioindicator species.

5. TRAINING OBJECTIVES 4. Given actual plants or high quality images, the trainee accurately scores the plants for injury amount and injury severity. 5. Given actual leaves and voucher materials, the trainee prepares a proper leaf voucher sample and data sheet. 6. Upon visiting an actual field site, the trainee prepares a site map that meets minimum standards outlined in the Field Guide.

6. OZONE TRAINING AGENDAMorning Session: inside 8:00 to 11:30 • Introduction - Why do we do it? • Field Procedures - How do we do it? • Symptom Review - What does ozone injury look like? • Written Test** - What information is in the Field Guide? • Electronic Data Entry and Data Sheets • Plot Location Check – State Crosswalk Tables LUNCH **requires a passing grade for certification

7. OZONE TRAINING AGENDAAfternoon Session: outside 1:00 to 4:00 • Field exercise - What does a biosite look like? • Review of species ID and plant selection • Review of symptom identification and scoring • Biosite map - How is biosite location documented? • Voucher handling - How are injury symptoms validated? • Review of QA activities • Debriefing Form • Distribution of field supplies

8. Stratospheric ozone is known as GOOD OZONE because it screens out harmful radiation from the sun. WHAT IS OZONE? Ozone in the troposphere is considered a pollutant (BAD OZONE) when ground-level concentrations exceed background levels of 10-40 ppb. Ozone pollution occurs under conditions of bright sunlight and warm temperature as a result of photochemical reactions involving nitrogen oxides and hydrocarbons emitted from cars and industry.

9. HOW IS OZONE FORMED? VOC + NOx + Sunlight = Ozone Ozone formation is a simple reversible reaction between oxides of nitrogen (NOx) and elemental oxygen (O2) driven by the energy of sunlight. The reversible reaction results in no net gain of ozone due to the instability of the O3 molecule. However, when VOC’s are present, they interfere with the reverse reaction resulting in a build-up of ozone in ambient air.

10. WHAT ARE THE SOURCES OF OZONE POLLUTION? Emissions from industrial facilities and electric utilities, motor vehicle exhaust, gasoline vapors, and chemical solvents are some of the major sources of the nitrous oxides (NOx) and volatile organic compounds (VOC) that lead to ozone pollution. The single most important source of ground-level ozone pollution is the car.

11. HOW DOES OZONE GET TO THE FOREST? Ozone is a regional scale pollutant. Long-range transport of ozone contaminated air masses within slow moving and often stagnant high pressure systems leads to high ambient ozone concentrations in forested areas downwind of urban centers. At 8:00 am commuters choke NY/NJ highways. At 11:00 am prevailing winds push car exhaust pollution northeast into CT By 3:00 pm the pollution cloud has moved into rural areas of MA and intense sunlight causes high ozone levels.

12. Ground-level Ozone volatile organic compounds + oxides of nitrogen + intense sunlight = O3 Relatively clean ambient air Air quality in Acadia National Park appears good in November and poor in August, when warmer temperatures and sunlight increase ozone formation. Rural areas often experience higher ozone concentrations than urban areas do, because ozone takes time to form. Winds carry ozone precursors (chemical products of fossil fuel combustion) away from urban areas where they originate, and ozone forms during that travel time. High ambient ozone concentrations

13. Development of a typical ozone episode in the Northeast region. At Day1 the air quality is good with ozone levels below 60ppb. By Day3, the air quality is moderate (yellow). By Day5, ozone levels are above 100ppb and unhealthy for sensitive groups. Air quality returns to background levels by Day8. Day 3 Day 1 Day 5 Day 8 A very unhealthy ozone day for the mid-Atlantic region. Ambient ozone concentrations are in the 125+ppb range, in exceedance of the National Ambient Air Quality Standard.

14. Ozone hot spots occur across the country downwind of populated areas. Normal wind and weather patterns tend to carry ozone into our region from urbanized areas to the south and west. Red areas are in violation of the national ozone standard set to protect plants and people from ozone injury.

15. Colored areas have a record of ambient ozone concentrations that are out of compliance with the National Ambient Air Quality Standard for Ozone.

16. The National Ozone Standards (NAAQS) 1-hour standards are met when the number of days each year with maximum hourly average ozone concentrations above 0.12 ppm is equal to or less than 1. 8-hour standards are met when the 3-year average of the annual 4th highest daily maximum 8-hour average concentration is less than 0.08 ppm. Key O3 Values in parts per million 0.04 = background (?) 0.06 = injury threshold for sensitive plants 0.08 = standard (1997 revision) – set to protect plant health 0.12 = older 1 hour standard in place since 1979 NEW O3 STANDARD 0.075 ppm March 12, 2008 EPA Contribution of the ozone biomonitoring program to national air quality policy: EPA Scientists and FIA Staff examined the ozone biomonitoring data and found a high percentage of U.S. counties where ozone injury to sensitive bioindicator species had been documented even though the counties were in compliance with the 1997 ozone standard. Conclusion: The ozone standard needed to be strengthened.

17. Revised Ozone AQI Breakpoints, 2008 For more information about EPA’s action to strengthen the national ozone standards: www.epa.gov/groundlevelozone.

18. Regional map: April 19, 2008 National map: June 9-10, 2008 This summer, New Englanders can expect an increase in the number of air quality alert days, resulting from EPA’s recent lowering of the level of the ground-level ozone air quality health standard. The new ozone standard is set at 0.075 parts per million (ppm) on an 8-hour average basis. EPA previously issued air quality alerts when 8-hour average ozone levels were at, or predicted to be at, 0.085 ppm or above. So far this year, there have been five days in New England when ozone concentrations have exceeded the new ozone standard.

19. Is there a relationship between ozone pollution and climate change? Tropospheric ozone is a potent greenhouse gas; Ozone is a major constituent of photochemical smog and part of the mix of greenhouse gases that contribute to climate warming. Because the atmospheric lifetime of ozone is 1 to 2 weeks in summer and 1 to 2 months in winter, ozone produced in a polluted region of one continent can be transported to another continent all year long. Figure: In 1860, there were no forests exposed to O3 > 60ppb. By the 1990’s, some 24% of global forests were exposed to O3 > 60ppb. The trend indicates a huge increase in forest areas at risk of ozone impacts. Monitoring needs are paramount.

20. HOW DOES OZONE INJURE PLANTS? Ozone enters the leaf through the stomata during normal gas exchange. Once ozone is inside, it attacks cell membranes and disrupts many physiological processes. If cells collapse and die than visible symptoms appear on the leaf surface. Upper-leaf surface where symptoms are visible. Lower leaf surface where ozone enters the leaf.

21. Upper-leaf surface Ozone Stipple

22. Environmental Conditions Affect Ozone Uptake and Resulting Plant Injury Biological Factors • Genetic diversity • Development stage Plant growth stage Leaf age Older, lower leaves are affected first

23. Environmental Conditions Affect Ozone Uptake and Plant Injury Physical Factors • Air movement • Sunlight • Temperature • Nutrition • Relative humidity • Soil moisture Any factor that influences whether the stomata are open or closed

24. Effects Of Ozone On Plant Growth Studies have shown that ambient ozone concentrations during the growing season can and do effect tree growth. For example: • Reduced photosynthetic activity • Changes in carbon allocation • Reduced height and diameter growth • Reduced viability of pollen and seeds • Altered water use • Predisposition to insects and disease • Visible damage to leaves and needles

25. Open-top fumigation chambers used in ozone effects research. EPA facility - Corvallis, OR.

26. More than 35 tree species have been identified as ozone sensitive based on foliar injury symptoms. In the field, symptoms are prevalent both on young trees and in the canopies of mature trees. Ponderosa pine - CA O3 sensitive black cherry in VT

27. Examples of O3 induced emergence tipburn and chlorotic dwarf on eastern white pine.

28. O3 haze O3 thin crown Bark beetle attack O3 chlorotic mottle

29. In the mixed conifer type in southern California, the elimination of ozone sensitive pines has led to an increase in ozone tolerant species characterized by thinner bark and branches close to the ground. This, in turn, presents a fuel ladder situation that jeopardizes the residual stand in the event of a catastrophic fire. Tree  ecosystem effect USDA FS Gen. Tech. Rep. 1996. Paul R. Miller and others

30. Genetic differences in O3 tolerance in black cherry genotypes. O3 induced leaf stipple O3 induced cell death sensitive intermediate tolerant

31. Genetic differences in O3 tolerance in trembling aspen genotypes. O3 induced bifacial necrosis sensitive intermediate tolerant Ozone stress predisposes aspen to attacks by leaf rust fungi. Karnosky and others.

32. FACE = Free-Air CO2 and/or O3 Enrichment System Northern Wisconsin - 1998 Aspen, Aspen/birch, Aspen/maple ecosystems Many cooperating scientists working on Global Climate Change Issues Study Assess the effects of elevated CO2 and elevated O3, alone and in combination, on height, diameter, and volume growth responses of 5 clones of aspen. Results All growth parameters were increased by elevated CO2 All growth parameters were decreased by elevated O3 No change in growth under elevated C02 plus O3 Conclusion Elevated O3 negates the expected positive growth effects of elevated CO2 in Populus tremuloides. Environmental Pollution Special Issue: Growth responses of Populus tremuloides clones to interacting elevated carbon dioxide and tropospheric ozone. 2001. Isebrands, McDonald, Kruger, Hendrey, Percy, Pregitzer, Sober, Karnosky.

33. Ecosystem-level Effect Diversity of an early successional plant community as influenced by ozone. 1998. Barbo, Chappelka, Somers, Miller-Goodman and Stolte. The ozone treatments = Ambient air (no chamber) Twice Ambient Air Carbon Filtered Air Non-Filtered Air Result = Canopy cover, vertical canopy density (layers of foliage), species richness, diversity, and evenness were all higher in Carbon Filtered Air than in all other treatments. Conclusion = Plant communities existing in areas where lower ozone concentrations are prevalent might be more complex and diverse than those existing in areas with higher ozone concentrations.

34. BIOINDICATOR PLANTS What is a bioindicator plant? Certain plants respond in a predictable and well-defined manner to above-background concentrations of ozone in the ambient air. These plants act as “detectors” of ozone pollution. Detection is based on a visible foliar response. Left: Chlorotic mottle on ponderosa pine. Center: Classic ozone stipple on sassafras. Right: Premature fall coloration on sweetgum.

35. BIOINDICATION Ozone bioindicator plants provide evidence of plant stress. They tell us not only that ozone concentrations were elevated for a particular time and place, but also that other necessary conditions for ozone uptake and injury (e.g., adequate light, nutrition, and soil moisture) were also present. The data collected on bioindicator plants provides FIA with information on ozone air quality and the potential for ozone impacts on forest growth and productivity over the long-term.

36. Why is the ozone indicator important? 1. TO MEET USFS INITIATIVES The USDA Forest Service made a commitment to the international community to monitor the area and percent of forestland subjected to levels of specific air pollutants, including ozone, which may cause negative impacts on the forest ecosystem. The FIA biomonitoring program is the only large-scale effort to monitor ozone stress in the natural environment. FIA Detection Monitoring and the1995 Santiago Agreement

37. Why is the ozone indicator important? 2. TO SUPPORT NATIONAL POLICY ON PLANT HEALTH PROTECTION There is a recognized need for biological data that can help inform and influence the establishment of meaningful air quality standards to protect plants from ozone damage. The scientific community says foliar injury data from natural systems is a priority research need. The FIA biomonitoring program meets that need.

38. Why is the ozone indicator important? • TO IMPROVE FOREST HEALTH - RISK ASSESSEMNT MODELS Model simulations are the only way we can get close to interpreting the risks associated with long-term ozone exposures. Biomonitoring data provides more, different, and better data that will improve the reliability of forest health models.

39. 3% reduction 16% reduction Model: Relationship between Ps and O3 for hardwood forests across the northeastern US, 1987-1992. O3 threshold = 0.04 ppm. Predicted result: Decrease in net primary production (NPP) from 3 to 16% as O3 dose increases. Greater percent reduction on soils with high water holding capacity.

40. Risk Assessment Models For Ozone And Forest Growth Growth Loss Percentage at Ambient O3 Levels Shafer and Heagle (1989) loblolly pine 5-26% Taylor (1994) loblolly pine - Teskey (1995) southern pines 2-9% Ollinger and others (1997) hardwood forests 3-22% Hogsett and others (1997) black cherry and aspen 20% yellow poplar and sugar maple 5-12% loblolly pine and white pine 5-12% Chappelka and Samuelson (1998) - 0-10% Largest uncertainty factor: use of seedling data from open-top chambers to estimate mature forest response.

41. ANALYSIS AND REPORTING Using the foliar injury data collected by the field crews, analysts calculate an index of ozone stress for each biomonitoring site. (BI = biosite index) Statistical techniques are applied to the index to generate a predicted response surface of ozone stress across the landscape.

42. Biosite Index and Risk Based on plant injury data from 1999 - 2002. The risk of probable ozone injury is low in the green areas, moderate in the yellow-orange areas, and high in the red areas.

43. National Ozone Risk Map Spatial interpolation of the mean biosite index for the 2000 to 2004 sampling period. Interpolated biosite index estimates are used to categorize the risk of probable ozone impact to forest ecosystems. Dark green indicates areas of relatively low risk, light green areas of moderate risk, and purple indicates areas of relatively high risk. Areas in white are not sampled.

44. STATE-LEVEL RESULTS SOUTH CAROLINA Biosite Index Categories on the X axis represent the risk of probable ozone injury to ozone sensitive tree species. In this example, over 10 million cubic feet of tree volume falls into the low to moderate risk zone in South Carolina and 53 percent of this total includes tree species that are ozone sensitive. However, low foliar injury scores coupled with below average rainfall indicate low O3 impact. Totals for each estimate

45. STATE-LEVEL RESULTS INDIANA Based on air quality and biosite data: 1. Indiana’s ozone exposures are high. 2. All forested acreage in the state is exposed to relatively high ozone. 3. Indiana crews routinely record high foliar injury scores. 4. The highest injury scores coincide with the highest ozone exposure levels in south central Indiana. Conclusion: Forests across the state are at risk of ozone-induced negative effects. SOUTH CAROLINA 1. South Carolina ozone exposures are moderate to high. 2. Low injury scores associated with several years of below average rainfall indicate low risk of ozone impacts to South Carolina forests. Map based on field crew data

46. Which map is better? Because the FIA Biosite Index is derived from plant response data, it has biological relevance. EPA air quality data from physical air samplers FIA air quality data from biomonitoring plots

47. Why Does the Biosite Index Work?

48. The Disease Triangle Each side of the triangle represents one of the three components of disease. Environment Total of conditions favoring ozone flux O3 = Pathogen Total of concentration and duration of exposure Amount of disease Severity of the foliar injury response Host Plant Total of conditions favoring susceptibility Acute exposure: O3 concentrations >80ppb for a few hours to days. Chronic exposure: O3 concentrations <60ppb for the entire life of a plant. Both acute and chronic exposures cause foliar injury and other adverse affects.

49. Each of the components of the Disease Triangle can display considerable variability, and as one component changes it affects the degree of disease severity within an individual plant and within a plant population. Soil moisture Air temperature Light intensity Nutrient availability Duration Frequency Concentration Environment Total of conditions favoring ozone flux O3 = Pathogen Total of concentration and duration of exposure Amount of disease Severity of the foliar injury response Host Plant Total of conditions favoring susceptibility Species Genotype Leaf age Leaf position in the canopy

50. Ozone and the Disease Triangle Concept A sensitive genotype, at a susceptible developmental stage, is exposed to elevated ozone concentrations in a predisposing environment for the minimum time period required for physiological dysfunction to occur. The end result is an ozone-induced diseased condition which may or may not lead to significant impacts on the forest ecosystem. Environment Total of conditions favoring ozone flux O3 = Pathogen Total of concentration and duration of exposure Amount of disease Host Plant Total of conditions favoring susceptibility