1. Results from the Agricultural Health Study Pesticide Exposure Study Linda Sheldon a, Kent Thomas a, Guadalupe Chapa a, Sydney Gordon b Martin Jones c, James Raymer d, Dale Sandler e, Jane Hoppin e Mustafa Dosemeci f, Aaron Blair f, and Michael Alavanja f a National Exposure Research Laboratory, U.S. Environmental Protection Agency b Battelle Memorial Institute, c University of Iowa, d RTI International; e National Institute of Environmental Health Sciences, f National Cancer Institute

2. 2 PRESENTATION OVERVIEW  Background of the Agricultural Health Study  Purpose of the Pesticide Exposure Study  Exposure Study Procedures  Applicator Exposure Measurement Results  Spouse and Child Measurement Results

3. 3 AGRICULTURAL HEALTH STUDY RESEARCHERS NCI and NIEHS are leading the epidemiological study and investigations of cancer and non-cancer health outcomes University of Iowa Department of Epidemiology operates the Iowa AHS Field Station Battelle Center for Public Health Research and Evaluation operates the North Carolina AHS Field Station Westat operates the AHS Coordinating Center EPA and NIOSH are leading AHS exposure sub-studies Battelle, University of Iowa, and RTI International conducted the AHS Pesticide Exposure Study

4. 4 AHS BACKGROUND The epidemiological study is designed to: Measure cancer and non-cancer health risks in the agricultural community Examine associations between use of agricultural chemicals, other exposures, and disease Determine factors that promote good health

5. 5 AHS PHASE I (1993 – 1997) Prospective cohort enrolled in Iowa and North Carolina: 52,395 private licensed pesticide applicators 32,347 spouses of private pesticide applicators 4,916 commercial applicators (IA only) Questionnaires collected information about pesticide use, work practices, other exposures, lifestyle factors, and health.

6. 6 AHS PHASE II (1998 – 2004) Follow-up through cancer registries and vital records linkage 5-year follow-up questionnaire via computer assisted telephone interview (CATI) Update health status, exposures, and lifestyle Buccal Cell Collection and Dietary Health Questionnaire Nested studies of exposure and specific health outcomes AHS PHASE III (2005 - 2008) Continued cancer and mortality follow-up Follow-up telephone interview Cross sectional and panel studies, nested case-control studies

7. 7 AHS RESULTS DISSEMINATION AHS Web Site www.aghealth.org Direct communication to study participants Fact sheets and other information distributed to and through Iowa and North Carolina Cooperative Extension Services Scientific and Informational Presentations Journal Publications

8. 8 The AHS is examining possible links between pesticide use and health risks Information on use of up to 50 common insecticides, herbicides, fungicides, and fumigants obtained from AHS enrollment and take-home questionnaires – current and historical uses Information on work practices collected from questionnaires AHS PESTICIDE EXPOSURE CLASSIFICATION

9. 9 2) Cumulative Exposure Years Used x Days/Year AHS EXPOSURE CLASSIFICATION 3) Adjusted Cumulative Exposure Years Used x Days/Year x Intensity Score 1) Ever/Never Use of a Pesticide

10. 10 Dosemeci et al., Ann Occ Hyg, 46:2, 2002 Intensity Score = (Mix + Appl + Repair) * PPE where, Mix: Mixing Frequency of Pesticides (0, 3, 9) Appl : Application Method (0 - 9) Repair: Repair of Application Equipment (0, 2) PPE: Personal Protective Equipment (0.1 - 1.0) AHS EXPOSURE INTENSITY ALGORITHM

11. 11 AHS PESTICIDE EXPOSURE STUDY Overall Goal Measure exposure to applied pesticides for a subset of AHS applicators to evaluate and improve the AHS exposure algorithm Specific Objectives  Measure exposure to target applied pesticides  Compare measurements to algorithm exposure intensity scores  Identify key exposure factors  Assess potential spouse/child exposure to farm- applied pesticides

12. 12 STUDY DESIGN OVERVIEW STUDY DESIGN OVERVIEW Target pesticides 2,4-D and Chlorpyrifos Eight exposure strata based on application method and PPE Applicators monitored on one day while using their normal procedures Observation of activities Dermal, personal air, urine samples Questionnaires after use Spouse and child biomarker measurement

13. 13 PARTICIPANT SELECTION AND RECRUITMENT Sample Frame AHS cohort members; completed AHS Phase I & II Reside in one of 22 Iowa or 22 NC counties Previous use of a target chemical Eligibility Screening Telephone screening Eligible with planned use of a target chemical Recruitment In-home visit to discuss study Applicator, spouse, child consent/assent Schedule monitoring visits

14. 14 OVERALL MONITORING TOTALS OVERALL MONITORING TOTALS IA NC Total Pesticide Applicators 84 a 24 108 Spouses (urine) 38 b 11 49 Children (urine) 9 3 12 a Includes 24 repeat monitoring visits for applicators b Includes 8 repeat monitoring visits for spouses

15. 15 NUMBER OF OBSERVATIONS IN EXPOSURE STRATA Broadcast Spray – Liquid 2,4-D or Chlorpyrifos No gloves, no enclosed cab 9 No gloves, enclosed cab 9 Gloves, no enclosed cab 14 Gloves, enclosed cab 21 In-Furrow – Granular Chlorpyrifos No enclosed cab 7 Enclosed cab 6 Hand Spray – Liquid 2,4-D No gloves 22 Gloves 20

16. 16 Visit 1 Provide urine sample containers; discuss scheduling Visit 2 Sample collection (dermal, air, urine) Observe activities Application questionnaire Visit 3 Sample collection (urine pick-up) Activity questionnaire SCHEDULE MONITORING SCHEDULE

17. 17 MEASUREMENTS Dermal Patches 10 Patches, sized proportionally to body area, under PPE and over clothing, combined for analysis Hand Wipe Combined wipes from 12 small areas on each hand Personal Air Personal monitor (pump and filter) Applicator Urine Pre-Application morning void Post-Application Composite (start of pesticide use through the following morning) Spouse and Child Urine Pre-Application morning void Post-Application morning void (2 days later)

18. 18 APPLICATOR MEASUREMENT RESULTS Comparison of exposure measurements across eight exposure strata Assessment of the relative contribution of hand loading, body loading, and air exposures to urinary biomarker levels Bivariate analyses for >80 pesticide use, work, and hygiene factor variables – selected results shown here

19. 19 Preliminary Results

20. 20 Preliminary Results

21. 21 Preliminary Results

22. 22 Preliminary Results

23. 23 Solid N = 13 Liquid N = 5 Ratio L/S Urine (post-application) u g/L a 10 b 323.2 Hand loading u g 1936018.9 Body loading u g 46021004.6 Personal air u g/m 3 0.7 1.0 a Measured as 3,5,6-trichloro-2-pyridinol (TCP) metabolite b Near average pre-application background levels of 9.8  g/L Applicator Chlorpyrifos Levels (Geometric Means) By Physical State (as applied) Preliminary Results

24. 24 Urine = β 1 Hand + β 2 Body + β 3 Air + Intercept + Error 2,4-DChlorpyrifos N8618 R2R2 0.560.35 p<0.00010.1015 Independent Variables Parameter Estimate p Parameter Estimate p Intercept1.210.02131.340.1045 HAND0.48<0.00010.120.3327 BODY-0.080.23430.150.4183 AIR0.200.01210.120.4936 Exposure Pathway Model Results Preliminary Results

25. 25 Hand Loading vs. Post-Application Urine Concentration (For liquid products only) Preliminary Results

26. 26 Broadcast Spray N = 49 Hand Spray N = 40 Ratio H/B Urine (post-application) u g/L 22* 43* 1.9 Hand loading u g 230** 1200** 5.2 Body loading u g 1200**19000**15.8 Personal air u g/m 3 0.26**1.1** 4.2 Applicator 2,4-D Levels (Geometric Means) By Application Method * 0.05 > p > 0.001 or **p ≤ 0.001 for significance of difference Preliminary Results

27. 27 Broadcast SprayHand Spray Significance Amount of active ingredient used * Hours active ingredient used *** Average wind speed ** Average temperature * Applicator Post-Application 2,4-D Urine Levels Regression with Selected Variables * 0.1 > p > 0.05 ** 0.05 > p > 0.001 *** p ≤ 0.001 Preliminary Results

28. 28 Broadcast SprayHand Spray YesNoYesNo Additive used (most often spreader or surfactant) 2619 126**31** Contact with sprayed vegetationn/a 165**30** Adjust nozzles during application27205831 Repair equipment192310134 Visible wind drift25185632 Minor spills, splashes, leaks, drips during mix/load 30186340 Minor spills, splashes, leaks, drips during application -a-a 206733 Applicator Post-Application 2,4-D Urine Levels (Geometric Mean ug/L) for Selected Variables a a Minimum number in any cell = 5 **p ≤ 0.01 for significance of difference Preliminary Results

29. 29 Broadcast SprayHand Spray NoYesNoYes Gloves worn during mix/load 51**16**82*25* Gloves worn during application 26** 7.8**71*22* Respiratory protection used, application 24*6.7* 43-a-a Long sleeves worn, application30175526 Head cover worn, application40184743 Applicator Post-Application 2,4-D Urine Levels (Geometric Mean ug/L) for Selected Variables a a Minimum number in any cell = 5 * 0.05 > p > 0.01 or **p ≤ 0.01 for significance of difference Preliminary Results

30. 30 Type of Footwear Used During Application Urine Hand Loading Body Loading Nug/Lug Chemically resistant10 34180049000 Leather26 40 80014000 Fabric or other6155 290018000 Hand-Spray Applicator 2,4-D Levels (Geometric Means) by Footwear Type Preliminary Results

31. 31 Cab VariableNUrine Hand Loading Body Loading Personal Air Unadjustedug/Lug ug/m 3 Open cab182139013000.23 Enclosed cab252621011000.25 Adjusted for AI a ug/L/kgug/kg ug/m 3 /kg Open cab183.81504800.086 Enclosed cab252.2351800.043 Broadcast Spray Applicator 2,4-D Levels (Geometric Means) by Tractor Cab Type a Measurement result divided by kg of pesticide active ingredient used Preliminary Results

32. 32 Cab Variable Urine Hand Loading Body Loading Personal Air Nug/Lug ug/m 3 Windows open 72527019000.42 Windows closed1923170 8600.20 No filter192619015000.29 Filter625300 4000.14 Broadcast Spray Applicator 2,4-D Levels (Geometric Means) by Enclosed Cab Variable Preliminary Results

33. 33 FAMILY MEASUREMENT RESULTS Measurement of urinary biomarkers for participating applicator family members Relationships between spouse and applicator urinary biomarkers Relationship between spouse 2,4-D urine levels and applicator work practices or household factors

34. 34 SPOUSE AND CHILD URINARY BIOMARKER MEASUREMENT RESULTS Geometric Means Ranges N Pre-App. Urine ug/L Post-App. Urine ug/L Post-App. Urine ug/L 2,4-D Spouse39 - 411.31.8< QL – 59 a Child91.62.0< QL – 5.9 Chlorpyrifos (TCP) Spouse6 - 87.64.3 1.1 – 8.3 Child35.53.1 1.3 – 5.3 a Spouse with highest measurement reported handling 2,4-D on monitoring day. Preliminary Results

35. 35 0.1 1 10 100 1000 0.11101001000 Applicator Urine (ug/L) Spouse Urine (ug/L) Spearman Correlation is 0.64, p < 0.0001 SPOUSE AND APPLICATOR POST-APPLICATION URINE 2,4-D CONCENTRATIONS Preliminary Results

36. 36 Observed Applicator Work N Median Spouse Urine 2,4-D ug/L p-value a Glove use during HML No103.2 0.03 Yes311.0 Minor spills, splashes, leaks, drips Yes83.90.01 No331.2 Adjust nozzles during application Yes103.8 0.01 No291.0 a Mann-Whitney U Non parametric comparison test Spouse’s Post-Application 2,4-D Levels based on Selected Applicator’s Work Practices Preliminary Results

37. 37 Reported Household ActivitiesN Median Spouse Urine 2,4-D ug/L p-value a Applicator’s removal of boots/shoes before entering home Yes232.1 0.11 No180.9 Spouse washed the applicator’s clothing in past week Yes291.70.56 No71.8 2,4-D use in the lawn or garden during past year Yes52.4 0.50 No361.7 a Mann-Whitney U Non parametric comparison test Spouse’s Post-Application 2,4-D Levels Based on Selected Household Activities Preliminary Results

38. 38 Home Distance FromN Median Spouse Urine 2,4-D ug/L p-value a Area where 2,4-D was mixed/loaded today <200 ft212.4 0.61 >200 ft181.7 Nearest area where 2,4-D was applied today <200 ft81.80.96 >200 ft321.9 Nearest area where 2,4-D was ever applied <200 ft321.9 0.73 >200 ft81.9 a Mann-Whitney U Non parametric comparison test Spouse’s Post-Application 2,4-D Levels Based on Home’s Distance from Pesticide Use Preliminary Results

39. 39 APPLICATOR MEASUREMENTS PRELIMINARY FINDINGS Large range in exposures for spray 2,4-D applicators in this study Lower exposures and range of exposures for chlorpyrifos applicators in this study Chlorpyrifos exposures for users of liquid products were much higher than those for users of granular products Significant differences in exposure were found between many exposure strata

40. 40 APPLICATOR MEASUREMENTS PRELIMINARY FINDINGS Differences in exposure for different application methods (hand spray > broadcast > in-furrow) Glove use was an important factor – wearing protective gloves reduced urine levels ~70% Role of enclosed tractor cabs in reducing exposures is less clear for mixer/loader/applicators Significant associations between urine and dermal or air measures – strongest for hand loading Several other factors appear to be important in increasing or decreasing exposures to 2,4-D

41. 41 SPOUSE AND CHILD PRELIMINARY FINDINGS Urinary 2,4-D levels were low but measurable for a majority of the spouses and children of 2,4-D applicators (geometric means  2 ug/L) One spouse reported directly handling a product containing 2,4-D and had the highest urine level (59 ug/L) Urinary TCP levels were measurable for the spouses and children of chlorpyrifos applicators – but these levels were similar to those found in non- farm populations

42. 42 SPOUSE AND CHILD PRELIMINARY FINDINGS Spouse and applicator post-application urine 2,4-D levels were significantly associated Some pesticide applicator work practices were significantly associated with spouse urine 2,4-D levels Some hygiene and household (laundering work clothing, removal of work boots, distance to fields) were not associated with significantly higher exposures to family members in this study Further study is needed with more people and for different chemicals

43. 43 COMMUNICATION OF RESULTS Journal articles in preparation Plan for development of AHS/PES outreach materials (brochure and slide sets) Audience AHS participants Pesticide safety educators Cooperative Extension Services Pesticide users

44. 44 ACKNOWLEDGEMENTS Researchers at several organizations provided key contributions to the AHS Pesticide Exposure Study design and implementation: EPA: Ruth Allen (OPP), Carry Croghan, Paul Jones AHS Federal Investigators: Cynthia Hines (NIOSH) NC AHS Field Station: Charles Knott and Joy Pierce (Battelle CPHRE) IA AHS Field Station: Charles Lynch and Ellen Heywood (University of Iowa) Field Study: Steven Reynolds (Colorado St. University), Gerald Akland (RTI International), Craig Hayes (North Carolina Department of Agriculture and Statistical Services) Sample Analysis: Marcia Nishioka (Battelle Columbus), Robin Helburn (RTI International), and David Camann (Southwest Research Institute) DISCLAIMER Although this work was reviewed by EPA and approved for publication, it may not necessarily reflect official Agency policy.