MATING DISRUPTION AND MONITORING OF CODLING MOTH. (Cydia pomonella L.) AND ORIENTAL FRUIT MOTH (Grapholita

Transcription

1 The Pennsylvania State University The Graduate School Department of Entomology MATING DISRUPTION AND MONITORING OF CODLING MOTH (Cydia pomonella L.) AND ORIENTAL FRUIT MOTH (Grapholita molesta Busck) IN PENNSYLVANIA APPLE ORCHARDS A Thesis in Entomology by Eric Bohnenblust Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science August 2009

2 ii The thesis of Eric Bohnenblust was reviewed and approved* by the following: Larry A. Hull Professor of Entomology Thesis Co-Advisor Grzegorz Krawczyk Senior Research Associate Extension Entomologist Thesis Co-Advisor Jim Schupp Associate Professor of Pomology Gary Felton Professor of Entomology Head of the Department of Entomology *Signatures are on file in the Graduate School

3 iii ABSTRACT The codling moth (CM), Cydia pomonella (L.) and oriental fruit moth (OFM), Grapholita molesta (Busck) have been identified as the cause for rejection of over 3500 loads of fruit from the eastern U.S. destined for fruit processing plants in Adams County, Pennsylvania since Recently, both OFM and CM have developed resistance to organophosphates and other commonly used insecticides that have been the major tools for control in the past. One new method to control the CM/OFM complex is through the implementation of mating disruption (MD) tactics. These studies focused on the interaction between monitoring trap placement height and MD dispenser placement height within the tree canopy. The efficacy of several new MD technologies was also investigated. The effectiveness of several dispenser densities of the CheckMate Duel MD technology was also evaluated in commercial apple orchards in Pennsylvania for CM and OFM. The goal of trap placement height versus dispenser placement height study was to understand the effect on moth captures between trap placement height and MD dispenser placement height. Traps were placed at a height of 1.8 m or 4.5 m in the tree canopy within plots that were 3 rows wide X 3 trees long. MD dispensers were placed at 3.2 m within the tree canopy in the MD treated plots while the other plots were treated with only insecticides. Capture of CM adults in monitoring traps was highest in traps placed at 4.5 m in the canopy in both treatment plots with and without MD. OFM adult capture in 2007 was lowest in traps placed at 1.8 m in plots treated with MD dispensers. In 2008, capture of OFM was highest in traps placed at 4.5 m in both MD and no MD treatments.

4 iv The goal of the MD efficacy study was to determine the effectiveness of several new MD technologies for control of CM and OFM when compared to the standard MD technology in Pennsylvania - Isomate CM/OFM TT, and insecticides only. The study consisted of replicated orchard plots about 2 ha in size of Isomate CM/OFM TT, TRE #9940 and Cidetrak OFM, and Disrupt Micro-Flakes for CM and OFM at their recommended rates. The Isomate CM/OFM TT technology was the most effective MD treatment for controlling CM. The TRE #9940 technology was also effective for CM control, while the Disrupt Micro-Flake technology was not as effective as the previous two MD technologies at reducing CM adult populations. Also, all three MD technologies reduced the number of CM captured in monitoring traps in 2008 from the number captured in The Cidetrak OFM, Isomate CM/OFM TT and Disrupt OFM Micro- Flake MD technologies were all effective at reducing OFM capture. The dispenser density study was intended to further our understanding of the effectiveness of several dispenser densities of the CheckMate Duel MD technology that is currently used by growers in apple orchards for the control of CM and OFM. The CheckMate Duel technology was placed at densities of 250, 375, and 500 dispensers/ha in replicated plots ( 2 ha) on several commercial farms in 2007 and In 2008, a replicated (plots 0.1 ha) trial with CheckMate Duel dispenser densities of 250, 375, 425, and 500 dispensers/ha was also placed in a commercial orchard. In 2007, the most effective dispenser density of the CheckMate Duel at reducing CM capture in monitoring traps was the low density of 250/ha. However in 2008, the CheckMate Duel dispenser density of 500/ha was the most effective density at reducing CM adult capture and live larvae in fruit. In the small plot study, the CheckMate Duel densities of 425/ha and

5 v 500/ha were the most effective densities for reducing adult capture of CM. The CheckMate Duel density of 250/ha did not shutdown adult OFM capture in monitoring traps as effectively as the higher dispenser densities in From these experimental studies, MD dispensers should be placed as high as possible in the tree canopy, while monitoring traps should also be placed high in the tree canopy but lower than the MD dispensers for effective monitoring of both CM and OFM in a MD environment. Also, the Isomate CM/OFM TT technology, and the combination of the TRE #9940 technology and the Cidetrak OFM can be used effectively by growers to control CM and OFM. The Disrupt Micro-Flake technology is currently only effective for the control of OFM, not CM. The effectiveness of MD increases the longer (i.e., across years) this tactic is used to control CM populations. Furthermore, the higher dispenser densities (425 and 500/ha) of the CheckMate Duel will have the greatest effect on both CM and OFM populations.

6 vi TABLE OF CONTENTS LIST OF FIGURES... ix LIST OF TABLES... xiv ACKNOWLEDGMENTS... xvi Chapter 1 LITERATURE REVIEW... 1 INTRODUCTION... 1 Sex Pheromones... 2 Mating Disruption... 3 Barriers to Insect Control by MD... 6 Factors Affecting MD Dispenser Placement... 8 Insect Pest Monitoring in Orchards Pheromone Trap Placement Chapter 2 EFFECT OF MONITORING TRAP PLACEMENT HEIGHT ON ADULT CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH [Grapholita molesta (Busck)] CAPTURE IN TRAPS BAITED WITH SEX PHEROMONE IN MATING DISRUPTION AND NON-MATING DISRUPTION APPLE ORCHARDS INTRODUCTION MATERIALS AND METHODS Orchard Sites Insecticides and Mating Disruption Trap Placement and Monitoring Codling Moth Virgin Female Traps Statistical Analyses RESULTS Moth Capture Trap Shutdown DISCUSSION Chapter 3 A COMPARISON OF VARIOUS MATING DISRUPTION TECHNOLOGIES FOR CONTROL OF CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH[Grapholita molesta (Busck)] IN APPLES INTRODUCTION MATERIALS AND METHODS Orchard Sites Insecticides... 37

7 vii Mating Disruption Products Disrupt Micro-Flake Counts Trapping and Monitoring Fruit Injury Evaluations Larval Identification Statistical Analysis RESULTS Moth Capture Fruit Injury and Larval Identification Micro-Flake Sampling DISCUSSION Chapter 4 A COMPARISON OF VARIOUS DISPENSER DENSITIES OF THE CHECKMATE DUEL FOR CONTROL OF CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH [Grapholita molesta (Busck)] IN APPLES INTRODUCTION MATERIAL AND METHODS Mating Disruption Technology Release Rate Analysis Experiment Site Selection Experiment Site Selection Experiments 1 and Insecticide Use Trapping and Monitoring Fruit Injury Evaluations Larval Identification Experiment Site Selection Insecticides Mating Disruption Technology Placement Trapping and Monitoring Statistical Analysis Experiment Experiment Experiment RESULTS Experiment Experiment Experiment Release Rate Analysis DISCUSSION... 76

8 viii Chapter 5 CONCLUSION LITERATURE CITED Appendix A Maps of Treatment and Trap Locations Appendix B Insecticide Schedules for 2007 and Appendix C Release Rate Analysis Data

9 ix LIST OF FIGURES Figure 2-1: Cumulative mean adult CM capture for first generation flight in traps baited with CM 1X lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars within each height with different letters are significantly different. Lower case letters correspond to 1.8 m height. T-test (P < 0.05) Figure 2-2: Cumulative mean adult CM capture for second generation flight in traps baited with CM DA Combo lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05) Figure 2-3: Cumulative mean adult CM capture for first generation flight in traps baited with CM 1X lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05) Figure 2-4: Cumulative mean adult CM capture for second generation flight in traps baited with CM DA Combo lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05) Figure 2-5: Cumulative mean adult OFM capture for the entire season in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars within each height with different letters are significantly different. Lower case letters correspond to 1.8 m only. T-test (P < 0.05) Figure 2-6: Cumulative mean adult OFM capture for the entire season in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. T-test (P < 0.05)

10 x Figure 3-1: Effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM 1X lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-2: The effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-3: The effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM 1X lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-4: The effect of three MD treatments on cumulative capture of adult CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-5: The effect of three MD treatments on the cumulative capture of adult female CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-6: The effect of three MD treatments on cumulative capture of adult female CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-7: The effect of three MD treatments on the mean cumulative adult CM capture in traps baited with CM 1X lures in 2007 compared to No MD is insecticides only. Within treatment means followed by the same letter are not significantly different. Tukey s HSD α = Figure 3-8: The effect of three MD treatments on mean cumulative adult CM capture in traps baited with CM DA Combo lures in 2007 compared to No MD is insecticides only. Within treatment means followed by the same letter are not significantly different. Tukey s HSD α = Figure 3-9: The effect of three MD treatments on the cumulative capture of adult OFM in traps baited with OFM lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α =

11 xi Figure 3-10: The effect of three MD treatments on the cumulative capture of adult OFM in traps baited with OFM lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Figure 4-1: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in pheromone monitoring traps baited with CM 1X and DA Combo lures in 2007 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Letters followed by an asterisk correspond to treatment followed by an asterisk. Tukey s HSD α = Figure 4-2: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in pheromone monitoring traps baited with CM 1X and DA Combo lures in 2008 Experiment 1. No MD is insecticides only. Treatments followed by the same letter are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Figure 4-3: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2007 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 4-4: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2008 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Figure 4-5: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in monitoring traps baited with CM 1X and DA Combo lures in 2008 Experiment 2. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 4-6: The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2008 Experiment 2. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α =

12 xii Figure 4-7: The effect of Isomate CM/OFM TT and four CheckMate Duel dispenser densities on the cumulative capture of adult CM in monitoring traps baited with CM DA Combo lures in 2008 Experiment 3. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Figure A-1: Map of treatment layout for MD technology comparison study in 2007 and The second insecticide only block for 2008 is not shown. Treatments are: insecticides only (solid black line), Disrupt Micro-Flakes (blue fill), TRE # 9940 and Cidetrak OFM (yellow long dash), Isomate CM/OFM TT (red short dash). Numbers correspond to replicates Figure A-2: Map of treatment layout in the CheckMate Duel dispenser study in Insecticides only blocks are not shown. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) Figure A-3: Map of treatment layout in the CheckMate Duel dispenser study in Insecticide only blocks are not shown. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) Figure A-4: Map of treatment layout in the CheckMate Duel dispenser study on the farm near Bendersville, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) and insecticides only (CV) Figure A-5: Map of treatment layout in the CheckMate Duel dispenser study on the farm near Gardners, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) and insecticides only (CV) Figure A-6: Map of treatment layout in the CheckMate Duel dispenser study on the farm near Peach Glen, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) and insecticides only (CV) Figure A-7: Example of trap locations within each treatment block for the MD technology comparison and CheckMate Duel dispenser density study Experiments 1 and 2 for both 2007 and

13 xiii Figure C-1: CM pheromone release rate of the CheckMate Duel and Isomate CM/OFM TT dispensers in ( ) indicate analysis performed by CBC America, Commack, NY or Suterra LLC, Bend, OR Figure C-2: OFM pheromone release rate of the CheckMate Duel and Isomate CM/OFM TT dispensers in ( ) indicate analysis performed by CBC America, Commack, NY or Suterra LLC, Bend, OR

14 xiv LIST OF TABLES Table 3-1: Percent CM/OFM injured fruit in all treatments in MD efficacy comparison study during Table 3-2: Percent CM/OFM injured fruit in all treatments in MD efficacy comparison study during Table 3-3: Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks Table 3-4: Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks Table 3-5: Number of Disrupt Micro-flakes visually detected at two different canopy heights during the initial inspection period after each application in 2007 and Table 3-6: Number of Disrupt Micro-flakes visually detected in the tree canopy during two week intervals from the date of first application Table 4-1: Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2007 Experiment Table 4-2: Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 Experiment Table 4-3: Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in experiment Table 4-4: Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in Experiment Table 4-5: Percent CM/OFM injured fruit in all treatments in the dispenser density study during farm 1, Experiment Table 4-6: Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 farm 2, Experiment Table 4-7: Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 farm 3, Experiment Table 4-8: Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in Experiment

15 xv Table B-1: 2007 insecticide schedule for the orchard used for the trap placement height vs. dispenser placement height study Table B-2: 2008 insecticide schedule for the orchard used for the trap placement height vs. dispenser placement height study Table B-3: 2007 insecticide schedule for the MD technology comparison study Table B-4: 2008 insecticide schedule for the MD technology comparison study Table B-5: 2007 insecticide schedule for the CheckMate Duel dispenser density study - Experiment Table B-6: 2008 insecticide schedule for the CheckMate Duel dispenser density study - Experiment Table B-7: 2008 insecticide schedule for the CheckMate Duel dispenser density study at the farm near Gardners, PA Experiment Table B-8: 2008 insecticide schedule for the CheckMate Duel dispenser density study at the farm near Bendersville, PA Experiment Table B-9: 2008 insecticide schedule for the CheckMate Duel dispenser density study at the farm near Peach Glen, PA Experiment Table B-10: 2008 insecticide schedule for the CheckMate Duel dispenser density study at the farm near York Springs, PA Experiment

16 xvi ACKNOWLEDGMENTS I would like to thank my advisors Dr. Hull and Dr. Krawczyk for their support and guidance throughout the last several years. I have learned much from both of them and I can only hope to be as successful as both of them in my future endeavors as a scientist. I would also like to thank Dr. Schupp for his guidance and thoughts throughout the entire process of performing this research. Also, I would like to thank my fellow students and friends, especially Neelendra Joshi, Dan Schmehl and Christy Harris for their help and willingness to listen and provide input and support. To my office-mates, Amanda Bachman and Lori Shapiro, thank you for putting up with me in our cubicle of an office throughout the course of the last several years. Also, to all of the summer interns at the Fruit Research and Extension Center, especially Greg Wenk, Matt Wagner and Andy May, thank you for all of your help carrying out much of the physical legwork through many long hot summer days. I would also like to thank my girlfriend Jackie. Without her support through many of the long arduous hours and her positive attitude this project would have been infinitely more difficult. My Mom and Dad were instrumental in their support of me throughout the last two plus years as well. I would also like to thank anyone that I did not mention above; your help was instrumental in the accomplishment of this project. Also, thank you to all of the people from Hercon Environmental for all of their assistance with the application of their products, material, and financial support. You were indispensable during the entire process. I would like to thank the many people from Suterra LLC for providing materials and financial support without which this project

17 xvii could not have been completed. Many thanks go out to everyone at Trece for providing materials for this project. Also, thank you to CBC America for providing materials to perform these studies. I also wish to acknowledge funding from the Pennsylvania Department of Agriculture (Grant ME ) and the fruit growers via the State Horticultural Association of Pennsylvania.

18 1 Chapter 1 LITERATURE REVIEW INTRODUCTION The codling moth (CM), Cydia pomonella (L.) and the oriental fruit moth (OFM), Grapholita molesta (Busck), are major pests of tree fruits in the United States. The CM/OFM pest complex in the eastern United States differs from that in the western U.S. because in the east, stone and pome fruits are more commonly planted in adjacent blocks on commercial farms. When pome and stone fruits are planted in close proximity to one another, OFM often infest pome fruits as well as stone fruits (Myers et al. 2006), while CM is a major pest of pome fruits only. In the western U.S., pome fruits are usually planted separately from stone fruits and OFM is currently not considered a major pest of pome fruits (Calkins and Faust 2003). Both CM and OFM feed internally within fruits rendering them unfit for human consumption. During the last several years, CM have accounted for the majority of rejected apple loads to processors (i.e., 1 load = bushels), thus becoming the largest insect threat to apples in Pennsylvania (Krawczyk pers. comm.). The CM/OFM complex has been identified as the cause for rejection of over 3500 loads of fruit from the eastern U.S. destined for fruit processing plants in Adams County, Pennsylvania since 1998 (Hull et al. 2008). Recently, both OFM and CM have developed resistance to organophosphate (OP) and other commonly used insecticides; the major tools for control of these pests in the

19 2 past (Krawczyk and Hull 2004; Krawczyk 2006). The emergence of pesticide resistance along with the ongoing OP phase-out mandated by the Food Quality Protection Act of 1996 has created a critical need for the development of new methods of control (Brunner et al. 2002). One of these new methods is the implementation of mating disruption (MD) tactics, which involves deploying synthesized sex pheromones, often in a large scale area-wide (AW) program across many adjacent orchards. Sex Pheromones Sex pheromones of Lepidopteran pests are generally blends of 10 to 18 carbonlong straight-chain primary alcohols, acetates, or aldehydes that are produced by adult females in order to attract males for mating (Baker and Heath 2005). In many moth species, the female calls males by releasing a specifically blended ratio of these molecules into the air (Carde and Haynes 2004). The pheromone blend, called a plume once it is released, is carried downwind by air currents where it is sensed by a male moth (Carde and Haynes 2004, Vickers 2006). This pheromone blend emitted by the female is species specific, and males are able to discriminate subtle differences between blends in order to prevent mating with the wrong species (Baker et al. 1998). Once the male moth has sensed the pheromone plume is the correct blend for his species, he orients upwind towards the source of the plume and begins casting side to side with forward surges in order to follow the pheromone plume to its source (Baker and Heath 2005). Once the male finds the source of pheromone, he lands near the female and begins courtship (Carde and Haynes 2004).

20 3 The sex pheromones of CM and OFM have been previously characterized. Roelofs et al. (1969) reported that the primary component of the OFM sex pheromone was (Z)-8-dodecenyl acetate. Later, Carde et al. (1979) found that the sex pheromone of OFM was actually four components that include (Z)-8-dodecenyl acetate, (E)-8- dodecenyl acetate, dodecanol, and (Z)-8-dodecen-1-ol. The CM sex pheromone was shown to contain (E)-8, (E)-10-Dodecandien-1-ol, also known as codlemone (Roelofs et al. 1971). Mating Disruption Mating disruption technology using sex pheromones can work via several different mechanisms. Currently, false plume following is believed to be one of the main mechanisms by which MD works (Bartell 1982, Miller et al. 2006b). In false plume following moths fly upwind and orient toward a pheromone dispenser instead of a female (Bartell 1982). False plume following can only occur when there is competitive interaction between MD dispensers and calling female moths (Bartell 1982). Recently, it has been found that higher numbers of MD dispensers that are widely spaced provide the best disruption of CM mating (Epstein et al. 2006). The higher number of dispensers increases the dispenser to female ratio which in turn reduces the probability that a male will find a female on their first attempt. The presence of mated CM females in sex pheromone treated orchards has been documented (Knight 2000, Light et al. 2001, Knight and Light 2005a); but the MD dispenser/female interaction reduces the probability that males will find females, and therefore can cause a delay in the mating of

21 4 female moths (Vickers 1997). It has been shown that delayed mating of female moths has a significant impact on the female s fecundity and fertility rate (Vickers 1997; Fraser and Trimble 2001; Jones and Aihara-Sasaki 2001; Jones et al. 2008). MD by competitive attraction has been reported to perform most effectively when pest populations are low to moderate (Miller et al. 2006b). Other proposed mechanisms through which MD may work are non-competitive. These include camouflage (Miller et al. 2006a), desensitization (Bartell 1982, Miller et al. 2006a) and sensory imbalance (Miller et al. 2006a). Camouflage is defined by a male being unable to locate a female because her pheromone plume has no boundaries and cannot be discerned from a background of ubiquitous synthetic pheromone (Miller et al. 2006a). Desensitization occurs when synthetic pheromone is released and male moths become less sensitive or responsive to the pheromone than they would be under normal conditions, and therefore be unable to locate and mate with a calling female (Miller et al. 2006a). Sensory imbalance works by confounding the males ability to perceive the necessary compounds that are associated with his species specific pheromone (Miller et al. 2006a). It has been proposed that these mechanisms may perform best when pest population densities are high (Miller et al. 2006b). Sex pheromones have been used successfully in the past to manage many different insect pests, including CM and OFM. One example is an area-wide MD (AWMD) program implemented in Washington State in 1994 to control CM in apples and pears (Calkins 1998). Pheromones have also been used successfully in Australia against OFM (Il ichev et al. 2002) as well as in Michigan on CM (Epstein et al. 2005) in AW programs. AWMD is more effective because when MD is applied to smaller blocks

22 5 of trees, mated female moths often enter the orchard from outside of the treated area (Calkins 1998). In AWMD orchards, mated female moths may enter on the boundaries causing some fruit injury, but the small ratio of outside edges to the inside area results in much better overall control (Calkins 1998). Currently, there is a large scale AW program in Pennsylvania which completed its third year of implementation in 2008 and has been successful in reducing fruit damage from the CM/OFM pest complex in both 2007 (Hull et al. 2008) and 2008 (Hull et al. 2009). A major benefit of MD is that it allows growers to reduce insecticide inputs, thereby reducing output costs and lowering risks to the environment and human health. It has been shown that MD works best when pest population densities are low to moderate. When there is a large population of the target pest, supplemental insecticides must be used along with MD to lower the population level in order for MD to be more effective (Pfeiffer et al. 1993; Brunner et al. 2002). The Washington State AW program was able to significantly reduce organophosphate insecticide inputs over the course of the program from six applications annually to an average of 0.7 applications annually (Calkins and Faust 2003). However, the overall ability to reduce pesticide inputs also depends on the need to control other pests present in the orchard such as Tortricid leafrollers (Knight 1995; Gut and Brunner 1998). The importance of reducing pesticide usage is a critical environmental and economic concern shared by fruit producers and the general public. Currently, MD dispensers cost around $300 per hectare ($120 per acre) plus application labor for a CM/OFM combination dispenser (i.e., Isomate CM/OFM TT or Checkmate Duel) (Hull pers. comm.). If this cost is added to a full insecticide program, it can reduce the economic feasibility of MD as a management technique.

23 6 The concurrent presence of CM and OFM in pome fruits in several fruit growing areas around the world has resulted in the development of MD products that contain both CM and OFM pheromone in the same dispenser (i.e., Isomate CM/OFM TT and Suterra CheckMate Duel). Previously, separate applications of two different dispensers were necessary to achieve control of both pests in pome fruit. This was a major obstacle to grower acceptance of MD because placing two dispensers in an orchard created high labor and material costs (Il ichev et al. 2007). The main pheromone dispenser used in apple orchards in the Pennsylvania AWMD project has been the Isomate CM/OFM TT product, because when the project was initiated in 2006, it was the only season long CM/OFM combination product available to Pennsylvania apple growers (Hull et al. 2007). However, in 2007, some growers began using the CheckMate Duel technology that was registered for use in Pennsylvania in early These two dispenser types were the main pheromone dispensers used for MD of CM and OFM in Pennsylvania during 2007 and Barriers to Insect Control by MD The use of MD has many potential positive aspects, such as reduced insecticide inputs for the targeted pests, increased biological control of pests, decreased opportunity for the development of insecticide resistance, and no reentry intervals for farm workers. However, there are also some barriers to overcome in order to achieve successful control of insect pests when using MD. One of these barriers is the edge effect at the borders of orchards. The edge effect is believed to be caused by male (Knight 2007) and mated

24 7 female (Calkins 1998) emigration from adjacent orchards. The edge effect is diagnosed by visibly increased levels of moth capture in pheromone traps and fruit injury at the border of an orchard when compared to the interior (Trimble et al. 1991; Pfeiffer et al. 1993; Gut and Brunner 1998; Il ichev et al. 2004). Several studies have found that the pheromone concentration on the upwind side of an orchard only becomes stable at least 15 m inside the orchard perimeter (Milli et al. 1997; Koch and Witzgall 2001), while on the downwind side pheromone concentrations similar to those present in the interior of the orchard have been recorded up to 60 m outside of the orchard (Milli et al. 1997). Karg and Sauer (1995) observed similar results in MD treated vineyards where pheromone concentration decreased significantly within 10 m outside of the vineyard edge. Knight (2007) reported higher CM captures in pheromone traps placed at the upwind edge of the orchard. Due to this edge effect, current monitoring protocols are to place traps at least 15 m from plot borders when determining the efficacy of a pheromone treatment (Stelinski et al. 2008; Stelinski et al. 2009). Currently, there are two main solutions to this edge effect; one is to apply additional insecticides to border rows in the orchard (Pfeiffer et al 1993; Knight 1995), and the second is to place extra dispensers around the outer edge of the orchard to increase the atmospheric pheromone concentration in this area. Another factor affecting MD efficacy is the pheromone release rate from the dispenser technology. Suckling et al. (1999b) found that adult Epiphyas postvittana (Walker) (light brown apple moth) captures decreased as atmospheric pheromone concentration increased in MD orchards. While, Stelinski et al. (2009) showed that OFM pheromone in Isomate CM/OFM TT and CM pheromone in Isomate CM/OFM/LR

25 8 dispensers were depleted faster than from dispensers that were specific to a single species such as Isomate CM TT and Isomate Rosso. They also noticed that the combination dispensers did not control fruit injury caused by CM/OFM as effectively as the single species dispensers after the dispensers were depleted of pheromone late in the growing season. Tomaszewska et al. (2005) showed that several dispensers loaded with varied amounts of codlemone released pheromone for at least 140 days post application, however, the authors did not quantify if the amount of codlemone released was sufficient to disrupt late season CM mating. It was also shown that different types of dispenser technologies have different rates of release (Tomaszewska et al. 2005). Microencapsulated sprayable pheromone formulations often have high release rates after application, but after several days the amount of pheromone released is not sufficient to prevent CM males from flying towards a pheromone source in the flight tunnel (Stelinski et al. 2005a). Carde et al. (1977) showed increased adult capture of CM late in the growing season in orchards treated with hollow fiber sources and attributed these captures to a lower rate of pheromone emission from the dispensers. In Italy, Angeli et al. (2007) found that Ecodian CP dispensers aged days in the field released pheromone at the same rate as a standard monitoring lure and dispensers aged 105 days released less pheromone than that of a monitoring lure. Factors Affecting MD Dispenser Placement Placement of MD dispensers within the tree and within the orchard are additional factors affecting the efficacy of MD treatments. Suckling et al. (1999a) found that

26 9 overall more pheromone is retained in the canopy when dispensers are placed lower, although pheromone concentration is greatest on the same plane as the dispenser. Several other studies have also found that pheromone concentration is highest on the same plane as the dispenser (Caro et al. 1980; Karg and Sauer 1995; Suckling et al. 1999b; Koch and Witzgall 2001). Based on these findings, they recommend that MD dispensers be placed where the targeted insect is most active. In the case of CM, the adults tend to be most active in the middle to upper part of the apple tree canopy (Borden 1931; Weissling and Knight 1995; Stoeckli et al. 2008). Rothschild and Vickers (1991) suggest that most OFM mating in peaches occurs high in the tree canopy, however, there have not been any definitive studies on the location of OFM mating behavior in the tree canopy in apples. Currently the industry recommendation is to place dispensers for CM (Weissling and Knight 1995) and OFM high in the canopy (Stelinksi et al. 2009). Also, pheromone concentration has been shown to be lower as the distance above a MD dispenser increases (Karg and Sauer 1995; Suckling et al. 1999a; Suckling et al. 1999b). Suckling et al. (1999a) suggests this is most likely due to increased scatter from environmental sources such as wind. This lower concentration also affects the need to place dispensers high in the canopy for an insect such as CM so that enough pheromone is present in their active zone. Suckling et al. (1999a) suggests that it may be necessary to compensate for this increased scatter in the upper canopy by placing an increased number of dispensers high in the canopy.

27 10 Insect Pest Monitoring in Orchards An important aspect of any successful control program for CM and OFM is the standardization of sex pheromone trap use for pest monitoring (Riedl 1980). Deployment of monitoring traps in orchards allows growers to more effectively assess the need for additional measures (i.e., application of insecticides) to control either CM or OFM. Sex pheromone monitoring traps have been used as population indicators to determine the necessity of insecticide applications in orchards since the 1970s (Madsen and Vakenti 1972; Vickers and Rothschild 1991; Knight and Light 2005b). Monitoring traps can also be useful in establishing biological reference points necessary to predict future points of an insect population s life cycle (Riedl et al. 1976; Vickers and Rothschild 1991). The use of biological reference points along with developmental models can help predict future phenological events that allow for more effective timing of insecticide applications (Riedl et al. 1976). In MD environments, moth capture in monitoring traps provides an indicator of the effectiveness of MD, because the lure acts as a calling female moth; capture of male moths in a pheromone trap indicates that males are likely able to locate females and mate. If no male moths are captured in the monitoring trap, then we assume that males are not able to easily locate and mate with females. However, it has been documented that CM can and do mate in MD environments (Knight 2006). One of the major limitations with sex pheromone monitoring traps is that they only capture male moths (Vickers and Rothschild 1991). Recently a new compound attractive to male and female CM has been discovered. Ethyl (2E, 4Z)-2, 4-decadienoate is a pear ester that acts as a kairomone and has the ability to attract both adult male and

28 11 female CM (Light et al. 2001). During field studies in California and Washington, Light et al. (2001) were able to capture female moths in traps baited with the pear ester in both walnut and apple orchards. Currently, there are two lures for use in monitoring traps that contain the pear ester, the DA Combo (Trece Inc., Adair, OK) lure that contains a 3.0/3.0 mg combination of codlemone, and the pear ester (Knight et al. 2005) and the DA (Trece Inc., Adair, OK) lure which contains only 3.0 mg of the pear ester (Knight and Light 2005a). In the western United States extensive work has been performed with the DA lure in attempting to establish a correlation between moth capture and possible fruit injury to determine the need for additional insecticide applications in MD environments (Knight 2002, Knight and Light 2005b). In Pennsylvania, the DA Combo lure has been shown to be more attractive to CM in MD environments than conventional sex pheromone lures (i.e., 1X lures) and performs as well as sex pheromone lures in non-md environments, while the DA lure is not effective at capturing CM in either environment (Joshi and Hull 2009). The DA Combo lure is also more attractive to male CM than females (Joshi and Hull 2009). The DA Combo lure could help provide a more accurate predictor of future potential fruit injury, since CM females oviposit on nearby leaves and fruit (Knight and Light 2004). However, more work still needs to be performed to correlate female moth capture with potential fruit injury.

29 12 Pheromone Trap Placement Several studies have examined adult CM capture in MD environments. One factor affecting adult capture rates is the placement of the pheromone trap in relation to a MD dispenser (Knight et al. 1999). However, this trap/dispenser interaction only appears to exist when traps are placed within 0.3 m of a dispenser (Knight et al. 1999). Barrett (1995) found that traps placed high in MD orchards captured the most CM. He also placed traps above the dispenser level; however, those traps were placed above the tree canopy and moth capture has been shown to be lower when traps are placed outside of the tree canopy than when traps are placed within the tree canopy (Riedl et al. 1979; Howell et al. 1990). Another factor that can affect CM capture is trap placement height within the tree canopy. CM activity has been observed to be highest in the upper third of the canopy (Borden 1931; Weissling and Knight 1995; Stoeckli et al. 2008), and the majority of studies have shown that pheromone traps placed in the top of the canopy capture the most CM (Riedl et al. 1979; McNally and Barnes 1981; Ahmad and Al-Gharbawi 1986; Barrett 1995; Tasin et al. 2008). Thwaite and Madsen (1983) found that when traps were placed in the same tree, the trap placed higher captured more CM than the trap placed lower. However, when they compared CM capture in trees where only one trap was placed low in the canopy versus trees with traps placed both high and low in the canopy, they found no difference in CM capture between the single trap and the sum of CM captured in both the high and low trap.

30 13 Many studies have also been performed to determine the optimal placement height for OFM monitoring traps. In non-md treated orchards, it has been shown that traps placed at a height of 1-2 m above the ground in the tree canopy capture more OFM than traps placed higher than 2 m above the ground (Beroza et al. 1973; Gentry et al. 1974; Rothschild 1975; Rothschild and Minks 1977). However, Evenden and McLaughlin (2004) found no difference in OFM capture in virgin female traps placed high in the canopy versus traps placed low in the canopy. De Lame and Gut (2006) found no differences in OFM adult capture in pheromone baited traps placed in the bottom third versus traps placed in the upper third of the tree canopy. In MD environments however, dispenser placement height affects the capture of OFM. When MD dispensers were placed in the top third of the tree canopy, no difference in adult moth capture was observed between monitoring traps placed high or low in the tree canopy. However, if the MD dispenser was not placed high in the tree canopy, traps baited with OFM pheromone placed high in the canopy tended to capture more adult moths (De Lame and Gut 2006). Atterholt (1996) also found that traps placed high in the canopy in MD treated almond orchards captured more OFM than traps placed lower in the canopy. In Pennsylvania s AWMD program, it was found that a number of CM monitoring traps were hung well above the dispenser in trees in commercial orchards (Hull pers. comm.). In other instances, limbs bearing fruit bent down as the fruit grew throughout the season, and if the limb contained a MD dispenser, the dispenser ended up lower than the CM monitoring trap. Only the work by Knight et al. (1999) and De Lame

31 14 and Gut (2006) reported on the relation between the position of an MD dispenser and the position of a monitoring trap within the tree canopy.

32 15 Chapter 2 EFFECT OF MONITORING TRAP PLACEMENT HEIGHT ON ADULT CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH [Grapholita molesta (Busck)] CAPTURE IN TRAPS BAITED WITH SEX PHEROMONE IN MATING DISRUPTION AND NON-MATING DISRUPTION APPLE ORCHARDS INTRODUCTION Sex pheromone monitoring traps have been used as population indicators to determine the necessity of insecticide applications in orchards since the 1970 s (Madsen and Vakenti 1972; Vickers and Rothschild 1991; Knight and Light 2005b). Moth capture in traps has also been used as a method to evaluate the efficacy of mating disruption treatments in orchards (Rothschild and Vickers 1991). Monitoring trap placement in orchards is of critical importance to effectively monitor codling moth (CM) populations (Knight et al. 1999) and oriental fruit moth (OFM) populations (De Lame and Gut 2006). Codling moth adults have been shown to be most active in the upper canopy of trees (Borden 1931; Weissling and Knight 1995; Stoeckli et al. 2008). Several studies have shown that when CM monitoring traps are placed high (4-5 m) in the tree canopy more adult moths are captured than when the traps are placed low (1.8 m) in the tree canopy (McNally and Barnes 1981; Ahmad and Al-Gharbawi 1986; Barrett 1995; Tasin et al. 2008). Barrett (1995) also found that traps placed high captured more CM in orchards treated with mating disruption (MD). Thus, the standard recommendation for monitoring has been to place CM traps high in the tree canopy (Barrett 1995).

33 16 OFM monitoring traps placed low in the tree canopy have been shown to capture adults as effectively as traps placed high in the tree canopy in conventionally managed orchards (Rothschild and Minks 1977; De Lame and Gut 2006). Several studies have reported that OFM capture is higher in traps placed at 1-2 m above the ground versus traps placed higher in the canopy (Beroza et al. 1973; Gentry et al. 1974; Rothschild 1975). In MD treated orchards, De Lame and Gut (2006) found that when dispensers were placed low in the canopy, OFM capture in traps that were placed high in the canopy was greater than OFM capture in traps placed low in the canopy. Atterholt (1996) also found OFM capture to be higher in traps placed high than in traps placed low in the canopy in almond orchards. Conventional recommendation for monitoring of OFM in apple orchards has been to place sex pheromone monitoring traps at 1.7 m in height in the tree canopy (De Lame and Gut 2006). In Pennsylvania, a number of monitoring traps for CM placed in MD treated apple orchards were found placed well above the dispenser placement height in trees (Hull pers. comm.). Also as the growing season progresses, the general shape of the trees change due to natural factors (i.e., fruit crop loads, tree growth), traps may not have the same proximal relationship to MD dispensers that they had early in the growing season. The goal of this research was to understand the effect of MD dispenser placement on CM and OFM adult capture in traps placed above and below the dispenser. This information should allow growers to monitor pests in their orchards managed under MD more effectively, producing fewer false negatives (the absence of trapped moths when damage is present). The increased effectiveness in monitoring should also allow growers

34 17 to more efficiently time insecticide applications in order to achieve cleaner fruit at harvest. MATERIALS AND METHODS Orchard Sites One 10 ha block of commercial apples in Biglerville, Pennsylvania was selected in 2007 and 2008 to evaluate the interaction between sex pheromone trap placement height and mating disruption dispenser height for CM and OFM. The block was divided into 16 plots of nine trees (3 rows X 3 trees) in 2007 and 24 plots of nine trees in Row and tree spacing in the block was 8.3 m and 5.1 m, respectively ( 270 trees/ha). Between each plot boundary there was a buffer of seven trees within each row and four adjacent rows that received only insecticides. The block consisted of four rows of Yorking followed by two rows of Golden Delicious cultivars in a repeating pattern. All trees were approximately m in height. Insecticides and Mating Disruption The grower s insecticide program was the same for both mating disruption and non-mating disruption treatments. All insecticides and application dates were set by the grower. Insecticides were primarily sprayed as alternate row middle (ARM) applications. The grower s insecticide schedules for 2007 and 2008 are shown in Appendix (B-1, 2).

35 18 The CheckMate Duel MD technology was provided by Suterra LLC, Bend, OR. CheckMate Duels are hand applied reservoir dispensers for both CM and OFM that release pheromone for the entire growing season. Each dispenser had two different pockets, one for the pheromone blend of each species. The pheromone blend for CM was (E,E)-8,10 Dodecadien-1-ol [17.54%] totaling 270 mg active ingredients per dispenser and other ingredients [82.46%], and the OFM blend was (Z)-8-Dodecen-1-yl acetate [11.93%], (E)-8-Dodecen-1-yl acetate [0.80%], (Z)-8-Dodecen-1-ol [0.15%] and other ingredients [87.12%] totaling 250 mg active ingredient per dispenser. The dispensers were applied to the upper part of the tree canopy with 3.1 m long wooden poles at a density equivalent to 375 dispensers per hectare and placed at a height of 3.1 m within each tree within each small MD treated plot. The second MD technology - Isomate CM/OFM TT - was provided by CBC America, Commack, NY. Each dispenser contained pheromone for both CM and OFM. The pheromone blend for CM was (E,E)-8,10 Dodecadien-1-ol [58.40%], Dodecanol [9.23%], tetradecanol [1.87%] totaling mg active ingredient per dispenser, and the blend for OFM was (Z)-8-Dodecen-1-yl acetate [21.25%], (E)-8-Dodecen-1-yl acetate [1.36%], (Z)-8-Dodecen-1-ol [0.23%] totaling mg active ingredient per dispenser and other ingredients [7.66%]. The Isomate CM/OFM TT dispensers are hand applied technology that emits pheromone for the entire growing season. The Isomate CM/OFM TT dispensers were applied to the upper part of the canopy with wooden poles at a height of 3.1 m and at a density equivalent to 500 dispensers per hectare. In 2007, half of the treatment plots received CheckMate Duel dispensers which were placed 5 May for the entire season and the other half received no MD treatment. MD and No MD treatments

36 19 were assigned using a completely randomized design. In the fall of 2007 after the leaves had fallen off of the trees, all dispensers still present in the tree canopy were removed from the orchard. During the 2008 season, eight plots received the CheckMate Duel dispensers, eight plots received Isomate CM/OFM TT dispensers both were applied 5 May for the entire season, and the other eight received no MD treatment. The CheckMate Duel, Isomate CM/OFM TT, and conventional insecticides treatments were assigned using a completely randomized design. Trap Placement and Monitoring In each small treatment plot one Pherocon VI white large plastic delta trap (LPD) (Trece, Inc. Adair, OK) for each species, CM and OFM, were both placed in the center tree. CM traps were placed on the east side of the tree and OFM traps were placed on the west side of the tree. In four of the plots within each treatment, the CM and OFM traps were placed at a height of 4.5 m and in the other four plots the traps were placed at a height of 1.8 m. Trap placement height was assigned using a complete randomized design. Traps for both species were placed at the same height within each tree. Rubber septa-style pheromone lures were used in all traps. For first generation CM in both 2007 and 2008, Pherocon CM 1X Long-Life lures containing only CM pheromone (i.e., codlemone) were used. For second generation CM flight during both years, the CM 1X lures were changed to Pherocon CM DA Combo lures loaded with codlemone and the kairomone - ethyl (E,Z)-2,4-decadienoate (pear ester) (Light et al. 2001). For OFM monitoring the Pherocon OFM 1X Long-Life lures containing only OFM pheromone

37 20 were used. OFM lures were replaced once per season in 2007 and The CM 1X lures were replaced with the CM DA Combo lures on 1 August in 2007 and on 1 July in The CM DA Combo lures were changed after seven weeks in Sticky bottoms in all traps were changed monthly. Traps were checked twice weekly and moths were counted and removed. In 2007, CM captured in traps baited with CM DA Combo lures were sexed by looking for the presence of claspers on their abdomen if male, while in 2008 CM males were determined by the presence of a black rectangle marking on the underside of the forewing (Fernandez et al. 2007). Codling Moth Virgin Female Traps In 2008, one Pherocon VI white LPD trap containing two CM virgin females in wire mesh cages was placed on 15 August in 20 plots consisting of nine trees (3X3) in the unused portion of the same orchard as the pheromone traps. Ten of the plots were treated with CheckMate Duel dispensers at a density of 375/ha and the other ten were treated with conventional insecticides only. In five of the plots treated with insecticides only and five plots treated with the CheckMate Duels, virgin female traps were placed in the canopy at 4.5 m. In the other five plots (10 total) within each treatment, traps were placed at a height of 1.8 m. There were five replicates of each trap height in the CheckMate Duel treated plots and in the insecticide only treated plots. All trap heights were assigned to plots using a completely randomized design. All virgin female traps were placed in the orchard between insecticide sprays to prevent females from being killed by insecticides. Traps were checked daily for captured male moths and female

38 21 survival. If the moths were dead, they were replaced if possible. Once the supply of newly emerged females was exhausted, traps in which females had perished were removed from the orchard. Statistical Analyses Cumulative moth capture data for each generation from pheromone traps for 2007 and 2008 was transformed [log(x+1)] and subjected to a two-factor (pheromone treatment and trap height) analysis of variance (Minitab 2003). The separation of means was determined using Tukey s test (Minitab 2003). When the interaction between the two factors was significant, a two sample t-test was performed on the log transformed moth captures for each factor separately (Minitab 2003). All tests were run at an α = 0.05 significance level. Trap shutdown is a measure of the rate of moth capture in sex pheromone monitoring traps in orchards treated with sex pheromone (i.e., MD) compared to orchards that are not treated with MD. Percent trap shutdown [(1-(no. moths captured in pheromone treated plot/no. moths captured in untreated plot)) X 100] can be used to measure the efficacy of sex pheromone mating disruption treatments (Rothschild 1975). Percent trap shutdown was calculated for each generation by averaging the percent shutdown for each trap height and treatment combination.

39 22 RESULTS Moth Capture The interaction between trap placement height and pheromone treatment on moth capture was very highly significant (F = 17.81; DF = 1, 12; P = 0.001) in traps baited with CM 1X lures for the first generation flight in 2007 (Fig. 2-1). There was no difference in cumulative moth capture between traps placed at 4.5 m (DF = 6; T = -1.28; P = 0.249) in CheckMate Duel or non-disrupted plots. CM capture in traps placed at 4.5 m was not different from capture in traps placed at 1.8 m in non-md plots (DF = 6; T = ; P = 0.249). Traps in plots treated with CheckMate Duel MD captured more CM at 4.5 m than at 1.8 m (DF = 6; T = 10.41; P = 0.000), while traps at 1.8 m in non-disrupted plots captured more CM than the traps in the CheckMate Duel treated plots at the same height (DF = 6; T = 4.29; P = 0.005) (Fig. 2-1). For the second generation flight of CM in 2007, adult CM capture was higher in traps baited with CM DA Combo lures placed at a height of 4.5 m than in traps placed at 1.8 m (F = 11.94; DF = 1, 12; P = 0.005) in both MD and non-md plots (Fig. 2-2). Moth capture was similar (F = 1.31; DF = 1, 12; P = 0.274) in plots treated with CheckMate Duel dispensers or insecticides only at both 4.5 m and 1.8 m (Fig. 2-2). During the first CM generation of 2008 more CM were captured in traps placed at 4.5 m than in traps placed at 1.8 m in plots treated with Isomate CM/OFM TT, CheckMate Duel and insecticides only (F = 88.91; DF = 1, 20; P = 0.000) (Fig. 2-3). Adult CM capture in CM 1X traps was higher in plots treated with insecticides than in plots treated with Isomate CM/OFM TT at both 4.5 m and 1.8 m (F = 3.48; DF = 2, 20; P = 0.05). CM capture in plots treated with CheckMate Duel dispensers was similar to

40 23 those treated with insecticides only or Isomate CM/OFM TT at either 4.5 m or 1.8 m. Higher CM capture during the second generation (F = 91.21; DF = 1, 20; P = 0.00) was also observed in traps placed at 4.5 m than at 1.8 m using CM DA Combo lures in Isomate CM/OFM TT, CheckMate Duel, and insecticide only treated plots in 2008 (Fig. 2-4). Adult CM capture was lower (F = 4.17; DF = 2, 20; P = 0.031) in plots treated with the CheckMate Duels at 4.5 m and 1.8 m than in plots that did not receive pheromone treatment at either height (Fig. 2-4). CM capture in plots treated with Isomate CM/OFM TT dispensers at 4.5 m and 1.8 m was similar to the number captured in the other two treatments at both heights. Male CM capture in traps baited with CM virgin females was not conclusive, as only two males were captured across all treatments. In 2007, there were no differences in adult female CM capture in traps baited with CM DAC lures placed at either height (F = 1.79; DF = 1, 12; P = 0.206) or in CheckMate Duel treated versus untreated plots (F = 0.02; DF = 1, 12; P = 0.880). There were also no differences in the capture of adult female CM at either height (F = 3.43; DF = 1, 18; P = 0.081) or in the Isomate CM/OFM TT, CheckMate Duel or insecticide only treated plots (F = 0.64; DF = 2, 18; P = 0.537) in A mean of less than two females were captured in each of the treatments during both years. In 2007, there was a significant interaction between trap placement height and pheromone treatment on cumulative OFM capture (F = 6.65; DF = 1, 12; P = 0.024) (Fig. 2-5). OFM capture was similar in traps placed at 4.5 m in CheckMate Duel treated plots

41 24 Adult CM per Trap a b A A No MD CheckMate MD 1.8 m 4.5 m Trap Height Figure 2-1 Cumulative mean adult CM capture for first generation flight in traps baited with CM 1X lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars within each height with different letters are significantly different. Lower case letters correspond to 1.8 m height. T-test (P < 0.05). Adult CM per Trap B B A A No MD CheckMate MD m 4.5 m Trap Height Figure 2-2 Cumulative mean adult CM capture for second generation flight in traps baited with CM DA Combo lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05).

42 A Adult CM per Trap C D CD B AB No MD Isomate MD CheckMate MD 1.8 m 4.5 m Trap Height Figure 2-3 Cumulative mean adult CM capture for first generation flight in traps baited with CM 1X lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05). Adult CM per Trap C CD D A AB B No MD Isomate MD CheckMate MD 1.8 m 4.5 m Trap Height Figure 2-4 Cumulative mean adult CM capture for second generation flight in traps baited with CM DA Combo lures in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P < 0.05).

43 a Adult OFM per Trap b A A No MD CheckMate MD m 4.5 m Trap Height Figure 2-5 Cumulative mean adult OFM capture for the entire season in Traps placed in Isomate and CheckMate Duel treated plots were placed below (1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars within each height with different letters are significantly different. Lower case letters correspond to 1.8 m only. T-test (P < 0.05). Adult OFM per Trap B B B A A A No MD Isomate MD CheckMate MD 1.8 m 4.5 m Trap Height Figure 2-6 Cumulative mean adult OFM capture for the entire season in Traps placed in Isomate and CheckMate Duel treated plots were placed below(1.8 m) and above (4.5 m) the height of the MD dispenser (3.1 m). Bars with different letters are significantly different. Tukey s HSD (P <0.05).

44 27 versus untreated plots at the same height (DF = 6; T = -0.15; P = 0.884). In nondisrupted plots cumulative OFM capture was also similar in traps placed at 1.8 m compared to traps placed at 4.5 m (DF = 6; T = 0.43; P = 0.681). More OFM were captured in traps placed at 1.8 m in non-md plots than in plots treated with the CheckMate Duels at the same height (DF = 6; T = -2.97; P = 0.025). In CheckMate Duel treated plots OFM capture was higher in traps placed at 4.5 m than in traps placed at 1.8 m (DF = 6; T = 5.07; P = 0.002) (Fig. 2-5). For 2008, more OFM were captured over the season in traps placed at 4.5 m when averaged across all treatments than in traps placed at 1.8 m within the tree canopy (F = 48.30; DF = 1, 18; P = 0.000) (Fig. 2-6). Similar numbers of adult OFM were captured in traps placed at either height in MD versus plots treated with only insecticides (F = 0.23; DF = 2, 18; P = 0.800). Trap Shutdown In 2007, the CheckMate Duel dispensers achieved a high (91.7 ± 8.3% [mean ± SE]) rate of shutdown in traps baited with CM 1X lures that were placed at 1.8 m, while adult CM capture in traps placed at 4.5 m was not reduced (-78.3 ± 37.8%) during the first generation flight. During the second generation flight, traps baited with CM DA Combo lures placed at 1.8 m in CheckMate Duel treated plots had a 70.9 ± 15.3% shutdown rate, while adult CM capture in traps placed at 4.5 m in CheckMate Duel treated plots was not reduced (-51.3% ± 28.2).

45 28 For the first generation flight in 2008, CM capture in traps baited with CM 1X lures placed at 4.5 m in CheckMate Duel treated plots was not shutdown (-10.9 ± 52.7%) while traps placed at 1.8 m had a 70.8 ± 15.3 % level of shutdown. The Isomate CM/OFM TT treatment shutdown CM capture at a rate of 26.7 ± 23.4% in traps baited with CM 1X lures placed at 4.5 m, while traps placed at 1.8 m were shutdown at a higher rate of 85.7 ± 14.3%. Percent shutdown during the second generation flight in traps baited with CM DA Combo lures that were placed at 4.5 m in plots treated with CheckMate Duel dispensers was 55.9 ± 12.9%, while shutdown of capture in traps placed at 1.8 m was higher (77.8 ± 14.7%). Also, a 42.0 ±15.3% level of shutdown was achieved in traps baited with CM DA Combo lures placed at 4.5 m in plots treated with Isomate CM/OFM TT, while moth capture at 1.8 m was almost completely shutdown (99.1 ± 0.9%). In 2007, the CheckMate Duel did not shutdown adult OFM capture in traps placed at 4.5 m (-56.4 ± 66.8%), however, traps placed at 1.8 m had a high (80.8 ± 15.9%) shutdown rate. Trap shutdown of OFM baited traps in 2008 was inconclusive due to large standard errors for all treatments. Shutdown of adult OFM capture in traps placed at 4.5 m in Isomate CM/OFM TT treated plots was not (-120 ± 156%) achieved, while traps placed at 1.8 m had a (36.9 ± 25.4%) shutdown rate. Shutdown of OFM capture in traps placed at 4.5 m in CheckMate Duel treated plots was also not (-4.9 ± 51.3%) achieved, while traps placed at 1.8 m had a (41.7 ± 30%) shutdown rate.

46 29 DISCUSSION In the first generation of 2007, traps placed at 1.8 m in CheckMate Duel treated plots captured fewer CM adults than traps placed at 1.8 m in non-md plots. Traps placed at 1.8 m in MD treated plots in both 2007 and 2008, had higher rates of shutdown than traps placed at 4.5 m in height in MD treated plots. The dispensers appear to be inhibiting the ability of CM to find pheromone monitoring traps when the trap is placed below the dispenser as evidenced by the high rates of trap shutdown and reduced captures of CM adults in MD plots. These results contradict the results of Knight et al. (1999) who reported that CM capture was greatest 1 m below a dispenser. This discrepancy could be a result of different experimental designs. Knight et al. (1999) do not mention tree size in their experiment and they only placed traps cm above a dispenser. There may not have been enough physical space in the tree canopy to place traps any higher. Also, they argued that there is a dispenser-trap interaction when the dispenser and trap are within 30 cm of each other, and if this interaction occurred in their experiment it could have affected CM capture above the dispensers in their study since their traps were placed within 30 cm of the dispenser. When the CM traps were placed above the MD dispensers, the percent trap shutdown was negative for both generations in 2007 and in the CheckMate Duel treated plots during the first CM generation of These findings demonstrate that if a monitoring trap is placed above a MD dispenser, the pheromone released by the MD dispenser will not consistently inhibit the males from finding pheromone monitoring traps or calling females. While the traps in plots treated with the Isomate CM/OFM TT

47 30 and CheckMate Duel dispensers captured significantly fewer moths at 4.5 m than the plots without MD during the first and second generation CM flights of 2008 respectively, the percent trap shutdown was low ( 27%) for the first generation flight and moderate ( 55%) for the second generation. Percent shutdown levels of only 27-55% indicate that adult males are more than likely still able to locate calling females and mate. If the dispenser is not placed in the top of the tree canopy, the dispenser headspace (atmospheric area above the dispenser containing a pheromone concentration) may not cover the entire portion of the canopy above the dispenser. Karg and Sauer (1995) found pheromone concentration decreases as distance above a dispenser increased in vineyards treated with MD for grape vine moth (Lobesia botrana Denis and Schiffermüller). Pheromone concentrations have been shown to be at their maximum on the same plane as the dispensers (Caro et al 1980; Karg and Sauer 1995; Suckling et al. 1999a; Koch and Witzgall 2001). Caro et al. (1980) recommended placing MD dispensers in the area where the insect is most likely to mate. In the case of CM, the adults tend to be most active in the middle to upper part of the tree canopy (Borden 1931; Weissling and Knight 1995; Stoeckli et al. 2008). My data supports these previous findings as well, based on the high shutdown rates for traps placed under dispensers, and the low rates of shutdown for traps placed above dispensers. Therefore, any increase in the overall pheromone concentration present within the tree canopy when a higher density of dispensers are placed in an orchard may not be enough to effectively disrupt CM in the upper canopy if the dispensers are not placed high in the canopy. The height of the monitoring trap in the tree was a significant factor affecting CM capture in plots treated with the CheckMate Duel during the first generation of 2007.

48 31 Traps placed high in the canopy in non-md plots, and above the dispenser in MD plots captured more CM than traps placed low in the canopy during the second generation of 2007 and both generations of 2008 (Fig. 2-2, 2-3, 2-4). These results are consistent with the findings of McNally and Barnes (1981); Ahmad and Al-Gharbawi (1986); and Tasin et al. (2008) that CM capture is highest in the upper part of the tree canopy in non-md orchards, and Barrett (1995) who showed CM capture to be highest in the top of the tree canopy in MD orchards. In the first CM generation of 2007, there was no difference in CM capture in traps placed high versus traps placed low in non-md plots. Interestingly, the effect of the MD treatment on CM capture was only significant for both generations in Female CM capture in CM DA Combo baited traps for both 2007 and 2008 was very low. The low capture could be a result of the pear ester present in the lures competing with other volatiles being released from ripening fruit (Mattheis et al. 1991) in the second half of the season that are attractive to CM females (Light et al. 2001). The traps baited with CM virgin females were ineffective at capturing male CM. This was likely due to the time during seasonal moth flight when the traps were placed in the orchard. Also, while moving about within their cages, the virgin females visibly damaged their wings and possibly other body parts which likely reduced their viability and their ability and willingness to call. Many studies have shown conflicting data about OFM capture within tree canopies. De Lame and Gut (2006) found that there was no difference in OFM capture in traps placed high or low in the tree canopy in plots without MD. Evenden and McLaughlin (2004) also found no difference in the effect of trap placement height on

49 32 OFM capture in traps baited with virgin females. While Atterholt (1996) found that OFM traps placed high in the tree canopy capture more OFM in almond orchards. Several other studies in apples have shown OFM capture to be greater in the lower part of the tree canopy (Beroza et al. 1973; Gentry et al. 1974; Rothschild 1975; Rothschild and Minks 1977). In my studies, OFM capture in 2007 was similar in traps placed at 1.8 m and 4.5 m in plots without MD. The 2007 results are similar to those found by Evenden and McLaughlin (2004) and De Lame and Gut (2006) but different from the other studies. During the 2008 season, OFM capture was higher in traps at 4.5 m than at 1.8 m in plots not treated with MD which is similar to the results found by Atterholt (1996). Possible explanations for the differences in OFM capture between my results and the previously cited studies that found higher OFM captures in the lower part of the canopy include tree height, range of heights monitored, and also one study was performed in peaches. Beroza et al. (1973) do not mention how tall the trees were in their study, and also they only placed traps at 0 m to 3 m in height. Rothschild and Minks (1977) conducted their study in peach orchards. It could be possible that different crops with different canopy structures affect the height where OFM are most active. The findings of Atterholt (1996) can likely be explained by the size of almond trees which are usually much larger than apple trees. Also, the trees in my study were tall (>5 m) and therefore the fruit was located further off the ground than in the study by Rothschild and Minks (1977) in peaches where the trees were only 3-4 m tall. The low captures of OFM in traps placed at 1.8 m in plots that were not treated with pheromones during 2008 could possibly be due to better insecticide control with several new compounds which would likely result in a reduced OFM population lower in

50 33 the tree canopy (Appendix B-2). Trees in this block were very tall ( 5 m) and there is more open area for the insecticides to penetrate the lower canopy more effectively than in the more dense upper canopy with an airblast sprayer. In 2007 and 2008, OFM capture was higher in traps placed at 4.5 m in height in MD treated plots suggesting that the dispensers did not fully inhibit the ability of male OFM to locate monitoring traps that were placed above the dispenser. This finding is similar to my results for monitoring CM. In 2007, OFM capture at 1.8 m in CheckMate Duel treated plots was lower than in untreated plots. This indicates that MD can effectively inhibit the ability of OFM males to locate a monitoring trap when the trap is placed below a dispenser. Trap shutdown for OFM was mostly inconclusive, the only case where the standard error of the mean (SEM) was not at least 25%, was in 2007 for traps placed at 1.8 m where there was a relatively high rate of trap shutdown. Otherwise, the SEM of percent shutdown for all other treatments was large and no viable conclusions can be based on percent shutdown for traps placed at 4.5 m in 2007 or any height in Some of the variation in percent shutdown in the OFM traps placed at 1.8 m in 2008 is likely due to the low moth captures at this height in plots not treated with pheromones. In conclusion, the placement height of a monitoring trap in relation to dispenser height is important; the most effective position for monitoring CM and OFM is high in the tree canopy. It is important that the dispenser be placed as high in the canopy as possible in order to provide the most effective disruption, while ensuring that dispensers are placed above the traps because the amount of pheromone in the headspace above a dispenser decreases with distance from a dispenser. Also, the dispenser should be placed

51 34 on a limb that is capable of carrying the harvest crop load without bending or breaking in order to prevent the trap-dispenser relationship from changing throughout the season. If the dispenser location does change over the season, the monitoring trap will not provide an accurate assessment of MD effectiveness because MD does not effectively disrupt the male moth s ability to find monitoring traps that are placed above the dispenser. Future research might address whether there is a difference in moth capture at different heights in relation to a dispenser in tall (i.e., 5 m) apple trees versus shorter (i.e., 2-3 m) trees. The canopy structure of a tree may affect the pheromone distribution differently, both within the tree and the orchard, in intensive orchard plantings where the tree canopies are more open than in taller trees with denser canopies in low density plantings. In addition, more research is needed on the efficacy of placing 50% of the MD dispensers in the lower 50% of the tree canopy and the other 50% of the dispensers in the top half of the canopy. When dispensers are placed at multiple heights within a tree, the pheromone released from the dispensers could possibly be more uniformly distributed throughout the entire canopy and be more likely to effectively shutdown adult captures in monitoring traps placed at any height within the tree canopy. Suckling et al. (1999a) showed that peak pheromone concentration within the tree declines as dispenser placement height increases. If half of the dispensers were placed lower it might be possible to maximize the amount of pheromone present in the tree and possibly more effectively disrupt CM and OFM.

52 35 Chapter 3 A COMPARISON OF VARIOUS MATING DISRUPTION TECHNOLOGIES FOR CONTROL OF CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH [Grapholita molesta (Busck)] IN APPLES. INTRODUCTION The codling moth (CM), Cydia pomonella (L.) and the oriental fruit moth (OFM), Grapholita molesta (Busck), are major pests of apples worldwide. Both CM and OFM feed internally within apples (Barnes 1991; Rothschild and Vickers 1991) rendering them unfit for processing or sale on the fresh produce market. In several areas of the world, CM and OFM occur in pome fruit orchards simultaneously during the growing season (Evenden and McLaughlin 2005, Il ichev et al. 2007, Hull et al. 2008). The CM/OFM complex has been identified as the cause for rejection of over 3500 loads of apples from the eastern U.S. destined for processing plants in Adams County, Pennsylvania since 1998 (Hull et al. 2008). Recently, OFM and CM have developed resistance to organophosphate (OP) and other commonly used insecticides that have been the major tools for control of these two pests in the past (Krawczyk and Hull 2004; Krawczyk 2006). The emergence of insecticide resistance along with the ongoing OP phase-out mandated by the Food Quality Protection Act of 1996 has created a need for new methods of control (Brunner et al. 2002). One method to control CM and OFM is sex pheromone mating disruption (MD) which has been shown to effectively reduce damage from CM and OFM in many parts of the world (Vickers et al. 1997; Gut and Brunner 1998; Brown and Il ichev 2000; Il ichev

53 36 et al. 2002; Angeli 2007). There are several products currently on the market for either CM or OFM, but currently Pennsylvania growers have only two MD options available for simultaneous season long CM and OFM control - Isomate CM/OFM TT and CheckMate Duel. The overall goal of this study was to compare the efficacy of several newly registered and experimental mating disruption products to the current industry standard in Pennsylvania - Isomate CM/OFM TT. This information will allow growers to make informed choices about the strengths and weaknesses of the Disrupt Micro-flake and Cidetrak products versus the products they are already using to control CM and OFM in their orchards. MATERIALS AND METHODS Orchard Sites Four apple blocks in Adams County near Aspers, PA on a single grower farm were selected to evaluate the following MD treatments: CM and OFM Disrupt Micro- Flakes from Hercon Environmental (Emigsville, PA), TRE #9940 and Cidetrak OFM from Trece Inc. (Adair, OK), and Isomate CM/OFM TT from America (Commack, NY) in 2007 and Insecticides were also applied in all MD treatment blocks. Each individual block comprised one replicate and was further divided into three smaller blocks between 1.52 ha and 3.32 ha in size. Each replicate block received one of the above treatments for CM and OFM (See Appendix A-1, 2 for the layout of treatment

54 37 plots). All four replicates were part of a larger area-wide mating disruption (AWMD) program and all treatment blocks were placed adjacent to each other. In 2007, one 4 ha block treated with insecticides only was used as a comparison block, and in 2008 another block ( 2 ha) treated with conventional insecticides only was added. All MD treatments except for the insecticides only treatments were assigned using a randomized complete block design. The insecticide only treatments were not blocked because the experiment was included in a larger area-wide mating disruption program. Apple cultivars were not standardized across the various blocks. Cultivars present on the farm were Delicious, Fuji, Gala, Ginger Gold, Golden Delicious, Ida Red, Nittany, and York Imperial. Insecticides All insecticides were chosen and applied by the grower. The grower primarily used alternate row middle applications to apply insecticides, but he did occasionally use complete (every row middle) sprays. The grower s insecticide schedules for 2007 and 2008 can be found in Appendix (B-3, 4). Mating Disruption Products Each Isomate CM/OFM TT dispenser contains a pheromone reservoir for CM and OFM. The pheromone blend for CM was (E,E)-8,10 Dodecadien-1-ol [58.40%], Dodecanol [9.23%], tetradecanol [1.87%], totaling mg active ingredients per dispenser (159.4 g a.i./ha) and the blend for OFM was (Z)-8-Dodecen-1-yl acetate

55 38 [21.25%], (E)-8-Dodecen-1-yl acetate [1.36%], (Z)-8-Dodecen-1-ol [0.23%] totaling mg active ingredients per dispenser (52.4 g a.i./ha) and other ingredients [7.66%]. The dispensers were applied at a density of 500/ha to the top 25% of the tree canopy. All Isomate dispensers were placed in the orchard between 5 and 10 May in 2007 and The TRE #9940 dispensers were applied to the top 25% of the canopy at a density of 1000/ha. The pheromone blend in the TRE #9940 dispensers was 75 mg of (8E, 10E) - 8, 10-dodecadien-1-ol and 110 mg of ethyl (2E, 4Z) 2, 4 decadienoate totaling 185 mg active ingredients per dispenser (185 g a.i./ha). The CideTrak OFM dispensers were applied at 425/ha ( g a.i./ha) to two blocks and 550/ha (137.5 g a.i./ha) to two other blocks in 2007, and all four blocks that received the Cidetrak OFM in 2008 were treated with a density of 425/ha. The pheromone blend in the CideTrak OFM dispensers was (Z)-8-Dodecen-1-yl acetate [4.65%], (E)-8-Dodecen-1-yl acetate [0.30%], (Z)-8- Dodecen-1-ol [0.05%] totaling 250 mg active ingredients per dispenser. All CideTrak OFM and TRE #9940 dispensers were placed in the orchard between 5 and 10 May in 2007 and The CM and OFM Disrupt Micro-Flakes were provided by Hercon Environmental (Emigsville, PA). The flakes were small laminated plastic squares (4 sq. mm) that were impregnated with sex pheromone. They were applied with a modified leaf blower provided by Hercon, Inc. that was strapped onto the bed of a John Deere Gator. The CM pheromone blend was (E, E)-8,10 Dodecadien-1-ol [5.97%] and other ingredients [94.03%]. This product was applied at a rate of 50 g active ingredient per hectare for each application. The OFM pheromone blend consisted of (Z)-8-Dodecen-1-yl acetate [7.83%], (E)-8-Dodecen-1-yl acetate [0.51%], (Z)-8-Dodecen-1-ol [0.08%] and other

56 39 ingredients [91.58%]. This product was applied at a rate of 19 g active ingredient per hectare for each application. The Disrupt Micro-Flakes were applied to all treatments on 15 May and 20 July in 2007, and on 15 May and 16 July in Disrupt Micro-Flake Counts Three weeks after the initial application of Disrupt Micro-Flakes for the first CM generation in 2007, eight random trees in each treatment blocks were selected to monitor flake numbers throughout the season. The eight trees were selected in two groups of four trees in each block and each grouping was located on opposite sides of the block. The same eight trees were used to monitor flake numbers after the initial application of the Disrupt Micro-Flakes in the second CM generation of 2007 and both CM generations of and Timed counts were taken for a total of six minutes per tree, 2 minutes in the upper 50% of the canopy, 2 minutes for the lower 50% of the canopy, and 2 minutes for a 1 m radius on the ground around the tree trunk. Flake counts were taken every two weeks from 8 June until the end of August in 2007 and 21 May until the end of August in Trapping and Monitoring Within each of the treatment blocks, three pheromone monitoring traps with OFM lures (Trece Long-life ) were placed in three replicates along with two traps each of Pherocon CM DA Combo (DAC) and CM 1X (Trece Long-life) lures (Trece, Inc,

57 40 Adair, OK). In the fourth replicate, only two monitoring traps containing OFM lures were placed along with the above combination of CM traps. CM traps were placed in each block in such a way that the lure pattern alternated DAC - 1X DAC- 1X. An example of trap layout is included in Appendix (A-7). All CM traps were placed on 2 m long bamboo poles and hung in the upper 20-30% of the canopy. OFM traps were placed at 1.8 m in height. Tree height ranged from about 3 to 5 m depending on the treatment replicate. All monitoring traps were checked weekly and the moths were counted and removed. CM adults captured in traps baited with the CM DAC lures were also sexed by looking for the presence of claspers if male, and an oviposition pad if female in In 2008 CM males were identified by looking for the presence of a black rectangle marking on the underside of the forewing (Fernandez et al. 2007). OFM lures were changed on 19 July 2007, and 17 July 2008, and CM 1X lures were changed on 12 July 2007 and 17 July 2008, and CM DAC lures were changed on 28 June and 9 August in 2007 and 26 June and 1 August in All trap floors were changed monthly. Fruit Injury Evaluations Estimates of fruit injury (i.e., apples with frass) by CM and OFM were taken three times during the 2007 season and twice (once at mid-season and harvest) during the 2008 season. These estimates were based on in situ evaluations of 100 fruit on 20 randomly selected trees in two separate rows within each treatment block. Samples were taken on 10 trees per row. Non-adjacent rows in the center of the block were chosen for sampling to minimize the effect of insects immigrating from adjacent blocks. At harvest, fruit

58 41 sampling was performed on trees of the cultivar Golden Delicious. If this cultivar was not present another cultivar was selected. Larval Identification Any apples which had visible frass during injury evaluations were collected and returned to the laboratory to be examined for the presence of live larvae. Larvae were preserved in a vial filled with KAAD (Bioquip Products, Gardena, CA) until they could be examined. CM larvae were identified by the lack of an anal comb and OFM larvae were identified by the presence of an anal comb. Statistical Analysis All cumulative adult capture data from MD treated plots for 2007 and 2008 was transformed [log (x+1)] and subjected to a two factor (treatment and block) analysis of variance (ANOVA). All moth capture data was analyzed from the date of the first full week of moth capture after MD dispenser placement. Cumulative adult CM capture in pheromone monitoring traps from 2007 and 2008 was transformed [log (x+1)] and subjected to an ANOVA to determine differences over time. Fruit injury data was also transformed log (x+1) and subjected to a two factor ANOVA. Moth capture and fruit injury data from the insecticides only treatment blocks was not analyzed because it was not part of the randomized complete block design. Mean data for all variables was separated using Tukey s HSD (α< 0.05) (Minitab 2003).

59 42 For the first assessment of Disrupt Micro-Flake distribution in the orchard, the counts were transformed [log(x+1)] and subjected to a two way ANOVA (height and block) to determine if there were differences in initial application height of the flakes. Within each generation, Disrupt Micro-Flake count data from all assessment times were also transformed [log(x+1)] and subjected to a two way ANOVA (time and block). Mean differences between each time period were separated using Tukey s HSD (α < 0.05) (Minitab 2003). RESULTS Moth Capture In 2007, cumulative CM capture in traps baited with CM 1X lures was higher in the Disrupt Micro-Flake treatment (F = 13.60; DF = 2, 18; P = 0.000) than in the TRE #9940 and Isomate CM/OFM TT treatments (Fig. 3-1). In traps baited with CM DAC lures, CM capture was reduced the most in the TRE #9940 and Isomate CM/OFM TT treatments (F = 10.02; DF = 2, 18; P = 0.001) (Fig. 3-2). CM capture in traps baited with both lure types was considerably lower in all MD treatments than in the insecticide only block. The CM population on this farm was extremely high in Traps baited with CM 1X and CM DA Combo lures captured about 900 CM and over 600 CM, respectively, in the insecticides only treatment. In the blocks treated with the Disrupt Micro-Flakes technology, nearly 200 and 150 CM were caught in traps baited with CM 1X and CM DA Combo lures, respectively.

60 43 In 2008, in traps baited with CM 1X lures, CM capture was again lowest in the the Isomate and TRE #9940 treatments (F = 7.10; DF = 2, 18; P = 0.005) (Fig. 3-3). CM capture was also lower in the Disrupt Micro-Flake treatment than in the insecticides only treatment (Fig. 3-3). In traps baited with CM DAC lures, CM capture in the Disrupt Micro-Flake treatment was higher than in the other two MD treatments (F = 12.55; DF = 2, 17; P = 0.000). There were few CM captured in the Isomate and TRE #9940 treatments and all MD treatments captured fewer CM than traps in the insecticides only treatment (Fig. 3-4). There was no difference in female CM capture between any of the MD treatments in 2007 (F = 2.86; DF = 2, 18; P = 0.075). In the insecticides only treatment, a cumulative mean of more than 12 females per trap was captured (Fig. 3-5). In 2008, there was again no difference in female CM capture between MD treatments (F = 3.06; DF = 2, 18; P = 0.084) (Fig. 3-6). In the insecticides only treatment, a cumulative mean of more than 7 female CM were captured over the 2008 season. Adult CM capture in all MD treatments in traps baited with CM 1X lures was also lower in 2008 than in 2007 (F = 21.97; DF = 1, 39; P = 0.000) (Fig. 3-7). In all MD treatments, traps baited with CM DAC lures captured fewer adult CM in 2008 than in 2007 (F = 12.73; DF = 1, 39; P = 0.001) (Fig. 3-8). In 2008, there was 76% fewer CM captured in traps baited with CM DAC lures and 61% fewer captured in traps baited with CM 1X lures in blocks treated with Isomate CM/OFM TT than in In blocks treated with Disrupt Micro-Flakes, CM capture was reduced by 62% in CM 1X traps and 74% in CM DAC traps from 2007 to The TRE #9940 treatment reduced the number of adult CM captured in CM 1X and CM DA Combo traps by 81% and 80%, respectively.

61 44 Adult CM capture in the insecticides only treatment was reduced by 28% and 53% in traps baited with CM 1X and CM DA Combo lures, respectively, from 2007 to OFM capture was similar in all MD treatments in both 2007 (F = 2.27; DF = 2, 27; P = 0.385) and 2008 (F = 2.27; DF = 2, 27; P = 0.422) (Fig. 3-9, 10). All three MD dispenser technologies were effective at completely shutting down OFM capture in OFM pheromone baited traps after their placement in the orchard. Fruit Injury and Larval Identification There was no difference in fruit injury between any of the MD treatments during the first (F = 1.00; DF = 2, 6; P = 0.422) and second (F = 1.00; DF = 2, 6; P = 0.422) mid-season evaluations in 2007 (Table 3-1). The insecticides only treatment block had 0.5% and 3.95% levels of fruit injury at the first and second mid-season evaluations, respectively. Fruit injury levels at harvest were similar between MD treatments in 2007 (F = 1.39; DF = 2, 6; P = 0.319). Fruit injury in the 2007 insecticides only treatment block was 19.1% at harvest (Table 3-1). In 2008, fruit injury was not different (F = 2.56; DF = 2, 6; P = 0.157) between any of the MD treatments during the mid-season evaluation (Table 3-2). There were no differences in fruit injury within any of the MD treatments at harvest (F = 2.05; DF = 2, 6; P = 0.209). Injury in the insecticides only treatment, however, was 0.35% at mid-season and 0.98% at harvest (Table 3-2). In 2007, few (< 5) live CM larvae were found in any of the MD treatments. However, during the harvest fruit injury evaluations, 29 CM larvae were found in the plot treated with insecticide only (Table 3-3). Few live larvae were found in any of the

62 45 treatments including the insecticides only treatment in 2008 (Table 3-4). All larvae found during both years were CM. Micro-Flake Sampling After the initial application of the Disrupt Micro-Flakes in May 2007, there were 76% more flakes detected in the bottom 50% of the tree canopy than in the top 50% of the tree canopy (DF = 1, 3; F =11.27; P = 0.044) (Table 3-5). There were no differences in the number of flakes in either half of the canopy after the second application in July 2007 (DF = 1, 3; F = 1.49; P =0.310), the first application in May 2008 (DF = 1, 3; F = 0.78; P = 0.442), or the second application in July 2008 (DF = 1, 3; F = 8.15; P = 0.065). The number of flakes present on the ground after each application was high (> 10) for all initial sampling periods except for the first application in This indicates that a large percentage of the flakes missed their target and did not adhere to the tree canopy. After the first application in 2007, the total number of flakes visually detected was higher during the third observation when compared to the first observation (DF = 2, 6; F = 13.60; P = 0.006) (Table 3-6). There were also more flakes visually detected in the lower portion of the canopy during the third observation than during the first and second observations (DF = 2, 6; F = 16.94; P = 0.003) (Table 3-6). There was no difference in the number of flakes detected in the upper 50% of the tree canopy during any of the observations (DF = 2, 6; F = 4.78; P = 0.057). There likely was not enough statistical power to conclude more flakes were observed during the third period. After the second application in 2007, the total number of flakes detected during either observation period

63 46 was similar (DF = 1, 3; F = 1.40; P = 0.322). After the first application in 2008, the number of total flakes visually detected during the different observation periods decreased during each sampling event (DF = 3, 9; F = 13.48; P = 0.001), most likely due to a rain event shortly after application. There were fewer flakes detected during the second, third and fourth sampling events. The number of flakes detected in the upper 50% of the tree canopy was lower during the third and fourth observation periods than the first observation point (DF = 3, 9; F = 9.37; P = 0.004) (Table 3-6). Flake numbers detected in the lower 50% of the tree canopy were lower during the second and fourth observation periods (DF = 3, 9; F = 6.83; P = 0.011) (Table 3-6). After the second application of 2008, the total number of flakes visually detected in the tree canopy was similar during all observation periods (DF = 2, 6; F = 0.11; P = 0.893). DISCUSSION The Isomate CM/OFM TT and TRE #9940 mating disruption technologies were the most effective products for reducing CM adult capture in monitoring traps baited with CM 1X lures for both 2007 and Disrupt Micro-Flakes were not as effective as the TRE #9940 and Isomate CM/OFM TT treatments at reducing CM capture in traps baited with CM 1X, or CM DA Combo lures in either year. In 2007 and 2008, adult capture in the Disrupt Micro-Flakes treatment was reduced when compared to the insecticides only treatment. These results are similar to those reported by Stelinski et al. (2008) who also found that the Disrupt Micro-Flakes were not as active in reducing CM capture when

64 Cumulative Mean Number of CM Per Trap A B No MD Isomate CM/OFM TT Disrupt Micro-Flakes TRE #9940 5/5 5/26 6/16 7/7 7/28 8/18 9/8 9/29 Figure 3-1 Effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM 1X lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Cumulative Mean Number of CM Per Trap /5 5/26 6/16 7/7 7/28 8/18 9/8 9/29 A B No MD Isomate CM/OFM TT Disrupt Micro-Flakes TRE #9940 Figure 3-2 The effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = 0.05.

65 Cumulative Mean Number of CM Per Trap A No MD Isomate CM/OFM TT Disrupt Micro-Flakes TRE # /2 5/23 6/13 7/4 7/25 8/15 9/5 9/26 B Figure 3-3 The effect of three MD treatments on the cumulative capture of adult CM in traps baited with CM 1X lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = CumulativeMean Number of CM Per Trap A B B No MD Isomate CM/OFM TT Disrupt Micro-Flakes TRE #9940 5/2 5/23 6/13 7/4 7/25 8/15 9/5 9/26 Figure 3-4 The effect of three MD treatments on cumulative capture of adult CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = 0.05.

66 49 Cumulative Mean Number of Female CM Per Trap /5 5/26 6/16 7/7 7/28 8/18 9/8 9/29 B B B No MD Isomate CM/OFM TT CM Disrupt Micro-Flakes TRE #9940 Figure 3-5 The effect of three MD treatments on the cumulative capture of adult female CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Figure 3-6 Cumulative Mean Number of Female CM Per Trap /8 5/29 6/19 7/10 7/31 8/21 9/11 10/2 No MD Isomate CM/OFM TT CM Disrupt Microflakes TRE #9940 The effect of three MD treatments on cumulative capture of adult female CM in traps baited with CM DA Combo lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = B B B

67 Cumulative Mean Number of CM Per Trap ± SEM No MD A B Isomate CM/OFM TT A B Disrupt Micro- Flakes A B TRE # Figure 3-7 The effect of three MD treatments on the mean cumulative adult CM capture in traps baited with CM 1X lures in 2007 compared to No MD is insecticides only. Within treatment means followed by the same letter are not significantly different. Tukey s HSD α = Cumulative Mean Number of CM Per Trap ± SEM No MD A B Isomate CM/OFM TT A B Disrupt Micro- Flakes A B TRE # Figure 3-8 The effect of three MD treatments on mean cumulative adult CM capture in traps baited with CM DA Combo lures in 2007 compared to No MD is insecticides only. Within treatment means followed by the same letter are not significantly different. Tukey s HSD α = 0.05

68 51 Cumulative Mean Number of OFM Per Trap /5 5/26 6/16 7/7 7/28 8/18 9/8 9/29 A* No MD Isomate CM/OFM TT* Disrupt Micro- Flakes* Cidetrak OFM 425/ha* Cidetrak OFM 550/ha* Figure 3-9 The effect of three MD treatments on the cumulative capture of adult OFM in traps baited with OFM lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Cumulative Mean Number of OFM Per Trap A* No MD Isomate CM/OFM TT* Disrupt Micro-Flakes* Cidetrak OFM 425/ha* 4/11 5/2 5/23 6/13 7/4 7/25 8/15 9/5 9/26 Figure 3-10 The effect of three MD treatments on the cumulative capture of adult OFM in traps baited with OFM lures in No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = 0.05.

69 52 Table 3-1 Percent CM/OFM injured fruit in all treatments in MD efficacy comparison study during Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/5/07 7/31/07 Harvest Isomate CM/OFM TT 0.01 (0.17) a 0.01 (0.17) a 0.03 (0.24) a Cidetrak OFM + TRE # (0.24) a 0.00 (0.00) a 0.04 (0.16) a CM and OFM Disrupt Micro-Flakes 0.08 (0.41) a 0.14 (0.56) a 1.19 (1.67) a Insecticides only 0.05 (0.00) 3.95 (0.00) (0.00) Means followed by the same letter are not significantly different. Table 3-2 Percent CM/OFM injured fruit in all treatments in MD efficacy comparison study during Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/13/08 Harvest Isomate CM/OFM TT 0.00 (0.00) a 0.03 (0.03) a Cidetrak OFM (0.01) a 0.00 (0.00) a TRE #9940 CM and OFM Disrupt 0.06 (0.05) a 0.10 (0.07) a Micro-Flakes Insecticides only 0.35 (0.35) 0.98 (0.78) Means followed by the same letter are not significantly different.

70 53 Table 3-3 Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks /5/07 7/31/07 Harvest Treatment CM OFM CM OFM CM OFM Isomate CM/OFM TT Cidetrak OFM + TRE # CM and OFM Disrupt Micro-Flakes Insecticides only Table 3-4 Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks /13/08 Harvest Treatment CM OFM CM OFM Isomate CM/OFM TT Cidetrak OFM +TRE # CM and OFM Disrupt Micro-Flake Insecticides only Table 3-5. Number of Disrupt Micro-Flakes visually detected at two different canopy heights during the initial inspection period after each application in 2007 and May application (±SEM) July application (±SEM) Top 50% 1.91 (0.26) a (0.84) a Bottom 50% 3.38 (0.73) b (3.40) a Ground* 4.47 (1.25) (6.44) 2008 Top 50% 5.06 (2.12) a 6.84 (1.22) a Bottom 50% 6.44 (3.09) a 9.66 (1.78) a Ground* (4.26) (3.69) Means after each application followed by the same letter are not statistically significant. *The number of flakes present on the ground was within a 1 m radius around the tree trunk and was not statistically compared to the flakes present in the canopy.

71 54 Table 3-6. Number of Disrupt Micro-Flakes visually detected in the tree canopy during two week intervals from the date of first application. Date st Application Top 50% Bottom 50% Total 6/8 1.91(0.26) a 3.38 (1.46) a 5.28 (0.99) a 6/ (0.49) a 4.53 (2.26) b 7.22 (1.59) a 7/5 3.16(0.71) a 7.19 (3.07) b (2.21) b nd Application 7/23* (4.17) a 8/9* (4.76) a st Application 5/ (2.12) a 6.44 (3.09) a (5.07) a 6/ (1.41) ab 3.09 (1.37) bc 6.16 (2.67) b 6/ (1.27) b 4.38 (1.96) ab 6.59 (3.09) b 7/ (0.93) b 2.41 (1.12) bc 4.19 (2.02) b nd Application 7/22* (2.85) a 8/5* (5.57) a 8/19* (3.14) a *If there were no significant differences in the total number of flakes between sampling periods then the means for the different heights are not shown because they were also not significant. Means within the same height level followed by different letters are not statistically significant.

72 55 compared to Isomate C Plus reservoir dispensers for CM, the standard MD technology used by many U.S. growers (Witzgall et al. 2008). All MD treatments in 2008 were effective at further reducing moth capture below 2007 levels. The reduction in moth capture from one year to the next shows that the effect of MD over time is cumulative and MD can provide increased control the longer it is used as a control tactic. Gut and Brunner (1998), and Hull et al. (2009), have also found an increased effectiveness of MD for control of CM and OFM over the course of several years during area-wide MD projects in Washington and Pennsylvania, respectively. It usually takes 1-2 years to reduce an insect population through the use of MD. Often, supplemental insecticides are necessary to prevent unacceptable levels of injured fruit in the first year until a population density can be reduced to a level where some insecticide applications can be eliminated (Gut and Brunner 1998). The number of CM females captured in traps baited with CM DA Combo lures was reduced in all MD treatments when compared to the insecticides only treatment in both 2007 and The reduced female capture in all three treatments during each year is interesting because the Isomate CM/OFM TT and Disrupt Micro-Flake treatments both contain only CM pheromone. The only MD treatment to contain both CM pheromone, and the pear ester that attracts female CM (Light et al. 2001) was the TRE #9940 formulation. The pear ester has been reported to stimulate oviposition by female CM (Knight and Light 2004) and the females are probably attracted to the kairomonal lures because they likely mimic a suitable fruit on which to lay their eggs. The pear ester was found to be a minor constituent of picked Delicious apples by Berger et al. (1984), however; the pear ester was not detected in the fruit analyses conducted by Light et al.

73 56 (2001). Another compound released by apples that has been reported to be attractive to female CM and stimulate oviposition is α-farnesene (Sutherland et al. 1977; Hern and Dorn 1999). The reduction in female captures in the Isomate CM/OFM TT and Disrupt Micro-flake treatments could be a result of a competitive attraction mechanism between the volatiles released from the large number of ripening fruit in the orchard (Light et al. 2001) and the CM DA Combo lure. Some of the possible apple volatiles that could be competing with the pear ester in the CM DA Combo lure are α-farnesene, and esters that are similar to the pear ester (Mattheis et al. 1991). This competitive attraction mechanism would then reduce the likelihood of the female finding a monitoring trap compared to the chances of finding a suitable fruit for oviposition. Capture of OFM in pheromone monitoring traps in all MD treatments was almost completely shutdown (capture of zero moths). Similar results have been obtained previously with several different types of OFM MD dispensers in Pennsylvania (Hull et al. 2001; Robertson and Hull 2002; Hull et al. 2007; Hull et al. 2008; Hull et al. 2009). It appears that all dispenser types were able to release enough OFM pheromone so that the pheromone level present in the orchard at any one time is higher than the threshold at which OFM can respond to pheromone point sources and find monitoring traps. The MD treatments along with supplemental insecticides reduced fruit injury caused by CM and OFM at harvest when compared to the 19.1% injury present in the conventional insecticides only treatment during In 2008, the Isomate CM/OFM TT and Cidetrak OFM/TRE #9940 and Disrupt Micro-Flake treatments along with supplemental insecticides (Appendix B-3, 4) reduced damage from CM and OFM at harvest when compared with the insecticides only treatment. These results are similar to

74 57 several other studies where fruit injury from CM and OFM was reduced through the use of MD and supplemental insecticides (Gut and Brunner 1998; Angeli et al. 2007; Il ichev et al. 2007; Lo and Cole 2007). No differences in fruit injury between MD treatments were present which indicates that the Disrupt Micro-Flake treatment while allowing increased moth capture in monitoring traps may still be effective at minimizing injury to fruit from CM and OFM. All of the damage that was present in this experiment was likely caused by CM, because no live OFM larvae were found during either year and OFM adult capture was low for the majority of each season. A possible explanation for the ineffective disruption of CM by the Disrupt Micro- Flake treatment could be that less CM pheromone is being applied initially coupled with a low proportion of flakes adhering to the foliage during the application process. Each application of the CM Disrupt Micro-Flakes is applied at 50 g a.i/ha per application (100 g a.i./ha for the whole season) while in the Isomate CM/OFM TT and TRE #9940 treatments, more than 159 g CM a.i./ha is initially applied. Of the total number of flakes counted in the tree canopy and in a 1 m radius on the ground around the tree trunk after each initial application, between 42 and 50 % of the flakes were on the 1 m radius around the tree trunk. It is also highly likely that many more flakes did not adhere to the tree canopy and landed on the ground in drive rows within the orchard increasing the likelihood that fewer than 50% of the flakes adhered to the foliage during the application process. Because CM are most active in the upper portion of the canopy (Borden 1931; Weissling and Knight 1995) and pheromone plumes are most concentrated on the same plane as the dispenser (Caro et al. 1980; Karg and Sauer 1995; Suckling et al. 1999b; Koch and Witzgall 2001), flakes on the ground are likely to have very little effect on the

75 58 disruption of CM mating in the tree canopy. Since such a low percentage of flakes actually adhere to the tree canopy, the amount of CM pheromone active ingredient present in the orchard after each application would be less than 25 g/ha. Meanwhile the initial amount of CM active ingredient present in the orchard in both the TRE #9940 and Isomate CM/OFM TT treatments was 185 g, and g/ha respectively. The difference in the amount of active ingredients per hectare indicates that there may not be enough pheromone present in the CM Disrupt Micro-Flake treatments to shutdown CM capture in monitoring traps. Another possible explanation for the increased adult captures in the Disrupt Micro-Flake treatments is that the individual flakes are less attractive than a CM 1X or DA Combo lure. Also, it could be possible that one of the main mechanisms through which these flakes achieve disruption is camouflage. Camouflage is defined as an adult male being unable to locate a female because her pheromone plume has no boundaries and cannot be discerned from a background of ubiquitous synthetic pheromone (Miller et al. 2006a; Witzgall et al. 2008). Stelinski et al. (2005a) have reported that camouflage is the likely mechanism through which mating disruption is achieved with sprayable pheromone formulations that are applied using airblast sprayers. Whereas, false plume following and habituation is thought to be the more common mechanism by which mating disruption is achieved through the use of reservoir dispensers (Epstein et al. 2005, Witzgall et al. 2008). However, Stelinski et al. (2009) argue that the mode of action for the Disrupt Micro-Flakes is competitive attraction without habituation. It could be possible that with the broadcast application of the flakes, the individual flakes begin by disrupting CM through false trail following and as the flakes age, the pheromone that is

76 59 being released after several weeks is insufficient to compete with females and traps, and therefore, ineffectively disrupting CM through the mechanism of camouflage. Ultra low volume (ULV) spraying is one method that has been used to increase the likelihood of sprayable pheromones to reduce CM captures with reasonable success (Knight and Larsen 2004). The sprayable formulations are generally characterized by many small droplets of pheromone that are more evenly distributed throughout the tree and appear to work as a mating disruptant via the mechanism of camouflage (Knight and Larsen 2004). When sprayable formulations are applied via the ULV, many droplets are concentrated in one area within the tree and possibly act similar to a single point source technology (i.e., hand applied dispenser), and thus likely utilizing the mechanism of false plume following and increasing mating disruption success (Knight and Larsen 2004). Applying the Disrupt Micro-Flakes in a similar spray pattern may increase their effectiveness. Also, the Disrupt Micro-Flakes did not appear to adhere to the foliage well during the first half of the season in 2008 (Table 3-6). It is possible that precipitation had an effect on the ability of the flakes to remain on the foliage. In 2008 there was a precipitation event (2.8 cm) over the hours (National Weather Service) following the first application of the flakes. It is likely that the glue which holds the flakes to the foliage was not set when the rain began and the flakes did not hold fast. Also, some of the glue might have worn off and caused the flakes to be less adhesive over time. Other possible explanations for the higher CM capture in the Micro-Flake treatment could be too little pheromone in the flakes to last a full CM generation or poor pheromone release rates. Data on the flake release rate in this study was unavailable.

77 60 However, if the rate that pheromone was released from the flakes quickly became insufficient to effectively disrupt the ability of CM to find traps; this problem might be partially solved through the use of ULV methods of application which would create a higher localized density of flakes. This localized density might offset the insufficient pheromone being released from a single flake and the combined release from all of the flakes in the localized area might create a longer lasting point source capable of disruption. Stelinski et al. (2009) showed that clusters of CM Disrupt Micro-Flakes function as attractive point sources when used as lures in monitoring traps. Also, increasing the amount of pheromone within an individual flake would likely increase disruption through sensory fatigue (Witzgall et al. 2008). If it is not possible to increase pheromone loading in individual flakes, increasing flake size would increase the amount of pheromone present in individual flakes. Both increasing the amount of pheromone in a flake and flake size may minimize the effect of a significant loss of pheromone from a dispenser immediately after application since a smaller percentage of the pheromone present in the dispenser would be lost. In conclusion, all three MD treatments were very effective for OFM control in apple orchards. However, the Disrupt Micro-Flakes treatment was not as effective as the TRE #9940 and Isomate CM/OFM TT treatments for control of CM. The TRE #9940 and Isomate CM/OFM TT dispensers both reduced CM capture in sex pheromone monitoring traps and prevented fruit injury at harvest. Future research of new methods to more efficiently apply the flakes, and also possibly concentrating the Micro-Flakes to create pockets of pheromone so the flakes affect moth capture similar to reservoir dispensers may improve the activity of this dispenser technology and increase the amount

78 61 of pheromone present in the orchard. Also, CM behavioral studies in the flight tunnel to determine the attractiveness of the Disrupt Micro-Flakes compared to monitoring lures should be conducted to further understand the lack of trap shutdown in orchards treated with this product.

79 62 Chapter 4 A COMPARISON OF VARIOUS DISPENSER DENSITIES OF THE CHECKMATE DUEL FOR CONTROL OF CODLING MOTH [Cydia pomonella (L.)] AND ORIENTAL FRUIT MOTH [Grapholita molesta (Busck)] IN APPLES. INTRODUCTION The codling moth (CM), Cydia pomonella (L.) and the oriental fruit moth (OFM), Grapholita molesta (Busck), are major pests of apples worldwide. Both CM and OFM feed internally within apples (Barnes 1991; Rothschild and Vickers 1991) rendering them unfit for human consumption. In Pennsylvania, both CM and OFM occur simultaneously in apple orchards (Evendon and McLaughlin 2005; Hull et al. 2009). Recently, OFM and CM have developed resistance to organophosphate (OP) and other commonly used insecticides that have been the major tools for control in the past (Krawczyk and Hull 2004; Krawczyk 2006). The emergence of insecticide resistance along with the ongoing OP phase-out mandated by the Food Quality Protection Act of 1996 has created a need for new methods of pest management (Brunner et al. 2002). One method to control CM and OFM that has been shown to effectively reduce damage to fruit in many parts of the world is mating disruption (MD) using sex pheromones (Vickers et al. 1997; Gut and Brunner 1998; Brown and Il ichev 2000; Il ichev et al. 2002; Angeli 2007; Hull et al. 2009). The concurrent presence of CM and OFM in pome fruits in several fruit growing areas around the world has resulted in the development of MD products that contain both CM and OFM pheromone in the same dispenser.

80 63 Several studies have found increased MD dispenser density to increase the effectiveness of CM (Epstein et al. 2006) and OFM (Stelinski et al. 2005b) control. However, hand applied dispensers are labor intensive to apply and therefore, placing more dispensers per unit area is more expensive to fruit growers. Suterra LLC (Bend, OR) recommended the CheckMate Duel dispenser for control of CM and OFM at a density range of /ha in 2007 and /ha in The goal of this experiment was to determine the lowest CheckMate Duel dispenser density that effectively disrupts mating and controls both CM and OFM. This information will allow Pennsylvania growers to use the most cost effective density of CheckMate Duels to manage CM and OFM in their orchards. MATERIAL AND METHODS Mating Disruption Technology The CheckMate Duel MD technology (Suterra LLC, Bend, OR) is a hand applied reservoir dispenser for mating disruption of both CM and OFM. Each dispenser had two different pockets, one for the pheromone blend of each species. The pheromone blend for CM is (E,E)-8,10 Dodecadien-1-ol [17.54%] totaling 270 mg active ingredients per dispenser and other ingredients [82.46%], and the OFM blend is (Z)-8-Dodecen-1-yl acetate [11.93%], (E)-8-Dodecen-1-yl acetate [0.80%], (Z)-8-Dodecen-1-ol [0.15%] and other ingredients [87.12%] totaling 250 mg active ingredient per dispenser.

81 64 The other MD product used in the experiments described below was the Isomate CM/OFM TT dispenser (CBC America, Commack NY). Each Isomate CM/OFM TT dispenser contains pheromone for CM and OFM. The pheromone blend for CM is (E,E)- 8,10 Dodecadien-1-ol [58.40%], Dodecanol [9.23%], tetradecanol [1.87%] totaling mg active ingredients per dispenser, and the blend for OFM is (Z)-8-Dodecen-1-yl acetate [21.25%], (E)-8-Dodecen-1-yl acetate [1.36%], (Z)-8-Dodecen-1-ol [0.23%] totaling mg active ingredients per dispenser and other ingredients [7.66%]. Release Rate Analysis Twenty trees in a commercial apple orchard were selected to determine pheromone release rate curves in Eight CheckMate Duel dispensers were placed in ten trees and eight Isomate CM/OFM TT dispensers were placed in ten other trees. Dispensers were placed in locations that received similar amounts of sunlight and rain. All of the CheckMate Duel and Isomate CM/OFM TT dispensers on a single tree were removed every two weeks for the entire season beginning on 6 May. Four each of the CheckMate Duel and Isomate CM/OFM TT dispensers were then shipped to CBC America, Commack, NY and the other half were shipped to Suterra LLC, Bend, OR for chemical analysis to determine the amount of pheromone remaining in each dispenser.

82 65 Experiment 1 Site Selection A commercial apple farm near Aspers, PA, was selected to compare the efficacy of the CheckMate Duel MD technology at three dispenser densities - 250/ha (67.5 g CM a.i./ha; 62.5 g OFM a.i./ha), 375/ha (101.3 g CM a.i./ha; g OFM a.i./ha), and 500/ha (135 g CM a.i./ha; 125 g OFM a.i./ha) compared to the Isomate CM/OFM TT at 500 dispensers/ha (159.4 g CM a.i./ha; 52.4 g OFM a.i./ha). All dispensers were applied to the upper 25% of the tree canopy and were placed in the orchard between 5 and 10 May in 2007 and In 2007 the farm was divided into three replicates based on the close proximity of various orchard blocks within each replicate. In 2008, the farm was divided into three replicates based on similar tree height. Each replicate was further divided into four smaller blocks of apples between 1.1 ha and 2.9 ha in size. Each smaller block within each replicate randomly received one of the four MD treatments (randomized complete block design) for CM and OFM (see Appendix A-2). All replicate apple blocks were part of a larger area-wide mating disruption (AWMD) program and all treatment blocks were placed adjacent to each other. The insecticide only treatments were not blocked because the MD treatments were included in a larger AWMD program. Tree height ranged from about 3 to 5 m depending on the replicate block. In 2007 and 2008, two 4 ha blocks that were located about 300 m from the MD blocks and treated with insecticides only were used for as no MD controls. Apple cultivars were not standardized across the entire farm and treatment blocks because the orchards had a

83 66 mixed cultivar planting. Cultivars present across this farm were Delicious, Empire, Gala, and Golden Delicious. Experiment 2 Site Selection In 2008, three commercial apple farms in Adams County, Pa, each near the towns of Peach Glen (farm 1), Gardners (farm 2), and Bendersville (farm 3), were selected to evaluate the same MD treatments as in Experiment 1 and an additional insecticide only control treatment. All dispensers were applied to the upper 25% of the tree canopy and were placed in the orchard between 7 and 17 May in Each farm was treated as a single replicate and all treatments were present within each replicate (A-4, 5, 6). Each treatment block (replicate) was between 1.6 ha and 2 ha in size. Tree height ranged from about 4 to 5 m depending on the replicate. No MD technology other than the above treatments was present anywhere on any of the farms. All treatments were assigned using a randomized complete block design. Apple cultivars were not standardized across the farms because the orchards consisted of mixed cultivar plantings. Cultivars present across the farms were Delicious, Empire, Gala, Golden Delicious, Granny Smith, Greening, Rome, and York Imperial.

84 67 Experiments 1 and 2 Insecticide Use All insecticides were chosen and applied by the grower, except that sprayable pheromone formulations were not used in any of the treatment blocks. The majority of insecticides were applied using the alternate row middle (ARM) method of application, but some insecticide sprays were applied as complete sprays. Insecticide schedules for each grower during 2007 and 2008 can be found in Appendix (B-3, 4, 5, 6, 7). Trapping and Monitoring Within each of the treatment blocks, three sex pheromone monitoring traps baited with OFM lures (Trece Long life ) were placed in two replicates along with two traps each of Pherocon CM DA Combo (DAC) and CM 1X (Trece Long life ) lures (Trece, Inc, Adair, OK). In the third replicate, only two monitoring traps containing OFM lures were placed along with the above combination of CM traps because the treatment blocks were small (< 1.8 ha). CM traps were placed in each block in such a way that the lure pattern alternated DAC - 1X DAC- 1X (A-7). All CM traps were attached to 2 m long bamboo poles and the trap was placed in the upper 30% of the canopy. OFM traps were placed at 1.8 m in height. All traps were checked weekly and the moths were counted and removed. CM adults captured in traps baited with the CM DAC lures in 2007 were also sexed by looking for the presence of claspers if male, or an oviposition pad if female. In 2008, CM males were determined by looking for the presence of a black rectangle

85 68 marking on the underside of the forewing (Fernandez et al. 2007). OFM lures were changed on 19 July, 2007, and 17 July, 2008, and CM 1X lures were changed on 12 July, 2007 and 17 July, 2008 and CM DAC lures were changed on 28 June and 9 August in 2007 and 26 June, and 1 August in All trap bottoms were changed monthly. Fruit Injury Evaluations Estimates of fruit injury (i.e., apples with frass ) caused by CM and OFM were taken three times during the 2007 season in Experiment 1 and twice (once at mid-season and harvest) during the 2008 season in both Experiments 1 and 2. These estimates were based on in situ evaluations of 100 fruit on 20 randomly selected trees in two separate rows within each treatment block. Samples were taken on 10 trees per row. Nonadjacent rows in the center of the block were chosen for sampling to minimize the effect from insects emigrating from adjacent blocks. At harvest, sampling was performed on trees of the Golden Delicious cultivar where possible. If this cultivar was not present another cultivar was selected. Larval Identification Any apples which had visible frass during injury evaluations were collected and returned to the laboratory and examined for the presence of live larvae. Larvae were preserved in a vial filled with KAAD (Bioquip, Gardena, CA) until they could be

86 69 identified. CM larvae were identified by the lack of an anal comb and OFM larvae were identified by the presence of an anal comb. Experiment 3 Site Selection One 40 ha apple block near York Springs, PA was chosen to test the same treatments as Experiment 2 and additionally a density of 425 dispensers/ha (114.8 g CM a.i./ha; g OFM a.i./ha) of the CheckMate Duel in The block was divided up into five replicates. Each treatment replicate consisted of one small square (9 trees long X 5 rows wide) plot of apple trees, approximately 0.1 ha in size ( 45 trees). Row and tree spacing in this orchard block was approximately 6.7 m X 4.3 m. Each small plot was separated by a buffer of 70 m of apple trees managed under the insecticide program that was applied to the entire orchard. All treatments were assigned using a randomized complete block design. The orchard planting consisted of Delicious, Golden Delicious, Rome, and York Imperial cultivars. Insecticides All insecticides were chosen and applied by the grower. The grower primarily applied complete sprays, but occasionally used ARM applications of insecticides. The insecticide schedule for 2008 is referenced in Appendix (B-10).

87 70 Mating Disruption Technology Placement Both the CheckMate Duel and Isomate CM/OFM TT dispensers were applied to the top 25% of the tree canopy and were placed in the orchard between 30 May and 2 June 2008 about 258 and 292 degree days after biofix (i.e. first sustained capture of CM adults in a pheromone monitoring trap). Trapping and Monitoring Monitoring was conducted as described in the previous experiments except that within each replicate 0.1 ha plot, one trap containing a CM DA Combo lure was placed in the center tree. Additionally, one trap containing an OFM lure was placed one tree to the west from the center tree at 1.8 m in height. Moth capture in all traps was zeroed (all moths captured previously were counted and removed) on 2 June to coincide with dispenser placement in all replicates. OFM lures were changed once 10 weeks after deployment and the CM DA Combo lures were changed 17 July, and 13 August. Trap bottoms were changed monthly. Statistical Analysis Experiment 1 Cumulative CM capture at the termination of moth flight, and cumulative female CM capture data was transformed log (x+1) and subjected to a two-way analysis of

88 71 variance (ANOVA). Cumulative OFM capture and fruit injury data was subjected to a two-way ANOVA. All data collected in the insecticides only treatment was not statistically analyzed because it was not part of the randomized complete block design. Means were separated using Tukey s HSD (Minitab 2003). All tests were performed at an α = 0.05 significance level. Experiment 2 Cumulative adult CM capture, and cumulative female CM capture in all treatments was transformed log (x+1) and subjected to a two way ANOVA. All cumulative OFM capture was subjected to a two-way ANOVA. Means were separated using Tukey s HSD (Minitab 2003). All tests were performed at an α = 0.05 significance level. Percent trap shutdown is a measure of the rate of moth capture in sex pheromone monitoring traps in orchards treated with sex pheromone compared to orchards that are not treated with pheromone (i.e., insecticides only). Percent trap shutdown [(1-(no. moths captured in pheromone treated plot/no. moths captured in untreated plot)) X 100] can be used to measure the efficacy of sex pheromone mating disruption treatments (Rothschild 1975). Mean cumulative percent trap shutdown for CM and OFM was calculated by averaging the percent trap shutdown for each MD treatment.

89 72 Experiment 3 Cumulative CM capture data was subjected to a two-way ANOVA. Cumulative female CM and OFM captures were transformed log (x+1) and then subjected to a twoway ANOVA. Mean separation was tested using Tukey s HSD (Minitab 2003). All tests were performed at an α = 0.05 significance level. Mean percent trap shutdown for male capture was calculated by averaging the percent shutdown for each treatment across all replicates. RESULTS Experiment 1 In 2007, total cumulative CM capture was lower in the blocks that received the Isomate CM/OFM TT treatment than in the blocks that received the CheckMate Duel dispenser densities of 375 and 500/ha (DF = 3, 41; F = 4.65; P = 0.007) (Fig. 4-1). In 2008 after randomization of the treatment replicates based on tree height, total CM capture was lower in the Isomate CM/OFM TT and the CheckMate Duel density of 500/ha treatment than in the CheckMate Duel density treatment of 250/ha (DF = 3, 40; F = 5.26; P = 0.004) (Fig. 4-2). CM capture in the intermediate CheckMate Duel density of 375/ha was not different from any of the other treatments. There was no difference in CM adult female moth capture in traps baited with CM DA Combo lures in any of the treatments in either 2007 (DF = 3, 18; F = 2.50; P = 0.092) or 2008 (DF = 3, 17; F = 2.45; P = 0.099) - data not shown. Cumulative adult female CM capture was low across all

90 73 treatments and ranged from moths per trap in 2007 and moths per trap in Adult OFM capture was similar in all MD treatments in both 2007 (DF = 3, 26; F = 0.94; P = 0.437) (Fig. 4-3) and 2008 (DF = 3, 26; F = 1.01; P = 0.405) (Fig. 4-4) while OFM were captured in the insecticides only treatments during both years. All MD treatments were effective at reducing OFM captures to near zero for the entire monitoring period during both seasons. Fruit injury caused by CM and OFM was not different between any of the MD treatments during the first mid-season evaluation (DF = 3, 6; F = 1.00; P = 0.455), the second mid-season evaluation (DF = 3, 6; F = 0.76; P = 0.555) or harvest evaluations (DF = 3, 6; F = 1.16; P = 0.398) in 2007 (Table 4-1). During 2008, there were again no differences in fruit injury levels between any of the MD treatments during the mid-season (DF = 3, 6; F = 1.00; P = 0.455) and harvest (DF = 3, 6; F = 1.13; P = 0.408) evaluations (Table 4-2). In 2007, seven live larvae were found during the harvest fruit injury evaluation in the plots treated with the CheckMate Duel densities of 375 and 500/ha, while few live larvae were found in any of the other treatments during any of the fruit injury assessments (Table 4-3). In 2008, again very few live larvae were found in any of the treatments during either of the fruit injury evaluations (Table 4-4). All larvae recovered during both seasons were identified as CM.

91 74 Experiment 2 Cumulative total adult CM capture was lower in all MD treatments than in the insecticides only treatment (DF = 4, 49; F = 12.35; P = 0.000) in 2008 (Fig. 4-5). Monitoring traps in the CheckMate Duel dispenser densities of 250/ha and 375/ha captured more CM than traps in the Isomate CM/OFM TT treatment. There was no difference in CM capture between the CheckMate Duel density of 500/ha and any of the other MD treatments. Adult CM female capture was not different across any of the treatments (DF = 4, 23; F = 2.11; P = 0.113) data not shown. Cumulative female CM capture was low across all treatments and ranged from per trap. Mean percent CM trap shutdown was moderate to high in the Isomate CM/OFM TT (82.3 ± 5.5%; Mean ± SEM), and CheckMate Duel dispenser density treatments of 375/ha (63.0 ± 4.63%) and 500/ha (73.0 ± 7.2%). However in plots treated with the lowest CheckMate Duel dispenser density (250/ha), only a low (15.8 ± 31.5%) rate of shutdown was achieved. Cumulative adult OFM capture was highest in non-md treated plots (DF = 4, 8; F = 14.89; P = 0.001) (Fig. 4-6). There was no difference in OFM capture between MD treatments. However, it should be noted that on one farm where a high population of OFM was present, OFM capture increased substantially during the last eight weeks of the season in the plot treated with the lowest CheckMate Duel density - 250/ha. Percent OFM trap shutdown was highest in the CheckMate Duel densities of 375/ha (99.9 ± 0.08), 500/ha (98.9 ± 0.07), and the Isomate CM/OFM TT treatment (98.0

92 75 ± 1.13). The CheckMate Duel density of 250/ha had the lowest (94.29 ± 3.23) rate of shutdown. Fruit injury was highly variable across farms (Table 4-5, 6, 7). The highest numbers of CM and OFM live larvae at harvest were found in the CheckMate Duel densities of 250 and 375/ha, and the insecticides only treatments (Table 4-8). Experiment 3 Total CM capture was significantly lower in plots treated with the Isomate CM/OFM TT technology and the CheckMate Duel densities of 425 and 500/ha (DF = 5, 20; F = 5.02; P = 0.004) than in plots treated with insecticides only (Fig. 4-7). The CheckMate Duel dispenser densities of 250 and 375/ha did not reduce total CM capture below that observed in the insecticides only treatment. There were no differences in the number of female CM captured in plots treated with either MD or insecticides only (DF = 5, 20; F = 1.06; P = 0.411) data not shown. Cumulative adult female CM capture averaged from females per trap for the season across all treatments. Also, no differences were observed in OFM capture in plots treated with MD or insecticides only (DF = 5, 20; F = 2.29; P = 0.085). Cumulative OFM capture in all treatments averaged less than two OFM per trap for the entire period of monitoring trap deployment, which is an extremely low population. The Isomate treatment achieved a moderate (63.2 ± 11.9%) percent CM trap shutdown rate. The CM shutdown rates for the various CheckMate Duel density

93 76 treatments were low to moderate and were as follows: 250/ha (39.1 ± 13.2%), 375/ha (42.3 ± 11.4%), 425/ha (48.4 ± 8.2%), and 500/ha (41.2 ± 15.1%). Release Rate Analysis Release rates during 2008 for CM pheromone from the CheckMate Duel and Isomate CM/OFM TT were similar and the dispensers continued to release pheromone for 140+ days (Appendix C-1). The CheckMate Duel released OFM pheromone effectively for about 60 days (Appendix C-2). After the first 60 days about 20% of the OFM pheromone was still present; after this period the release rate of the OFM pheromone from the CheckMate Duel slowed considerably. Based on the analysis from both companies, the CheckMate Duel only released between 3% and 10% of the OFM pheromone that was initially loaded in the dispenser over the last 80 days of the season. The OFM pheromone release rate from the Isomate CM/OFM TT dispenser was more constant than the rate of release from the CheckMate Duel dispenser for about days. DISCUSSION In Experiment 1 during 2007, the CheckMate Duel dispenser density of 250/ha was the only treatment that did not allow increased CM capture when compared to the Pennsylvania industry standard MD product - Isomate CM/OFM TT. These results contradict previous studies where higher dispenser densities were found to more

94 77 80 Mean Cumulative CM +/- SEM BC BC* AC A No MD Isomate CM/OFM TT CheckMate Duel 500/ha CheckMate Duel 375/ha* CheckMate Duel 250/ha 5/5 5/26 6/16 7/7 7/28 8/18 9/8 Figure 4-1 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in pheromone monitoring traps baited with CM 1X and DA Combo lures in 2007 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Letters followed by an asterisk correspond to treatment followed by an asterisk. Tukey s HSD α = Mean Cumulative CM +/- SEM B AB A* No MD Isomate CM/OFM TT* CheckMate Duel 500/ha* CheckMate Duel 375/ha CheckMate Duel 250/ha 5/2 5/23 6/13 7/4 7/25 8/15 9/5 9/26 Figure 4-2 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in pheromone monitoring traps baited with CM 1X and DA Combo lures in 2008 Experiment 1. No MD is insecticides only. Treatments followed by the same letter are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = 0.05.

95 78 45 Mean Cumulative OFM +/- SEM B B BB No MD Isomate CM/OFM TT Checkmate Duel 500/ha Checkmate Duel 375/ha Checkmate Duel 250/ha 5/5 5/26 6/16 7/7 7/28 8/18 9/8 Figure 4-3 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2007 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = Mean Cumulative OFM +/- SEM B* No MD Isomate CM/OFM TT* CheckMate Duel 500/ha* CheckMate Duel 375/ha* CheckMate Duel 250/ha* 4/11 5/2 5/23 6/13 7/4 7/25 8/15 9/5 9/26 Figure 4-4 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2008 Experiment 1. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = 0.05.

96 79 Table 4-1 Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2007 Experiment 1. Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/5/07 7/31/07 Harvest Isomate CM/OFM TT 0.00 (0.00) a 0.05 (0.04) a 0.18 (0.16) a CheckMate Duel 500/ha 0.03 (0.03) a 0.13 (0.09) a 0.72 (0.53) a CheckMate Duel 375/ha 0.13 (0.13) a 0.03 (0.03) a 0.73 (0.53) a CheckMate Duel 250/ha 0.00 (0.00) a 0.02 (0.01) a 0.03 (0.03) a Insecticides only 0.00 (0.00) 0.00 (0.00) 0.08 (0.08) Means followed by the same letter are not significant. Table 4-2 Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 Experiment 1. Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/13/08 Harvest Isomate CM/OFM TT 0.00 (0.00) a 0.02 (0.02) a CheckMate Duel 500/ha 0.00 (0.00) a 0.03 (0.03) a CheckMate Duel 375/ha 0.02 (0.02) a 0.05 (0.03) a CheckMate Duel 250/ha 0.02 (0.02) a 0.10 (0.05) a Insecticide only 0.00 (0.00) 0.05 (0.05) Means followed by the same letter are not significant. Table 4-3 Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in Experiment /5/07 7/31/07 Harvest Treatment CM OFM CM OFM CM OFM Isomate CM/OFM TT CheckMate Duel 500/ha CheckMate Duel 375/ha CheckMate Duel 250/ha Insecticide only

97 80 Table 4-4 Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in Experiment /13/08 Harvest Treatment CM OFM CM OFM Isomate CM/OFM TT CheckMate Duel 500/ha CheckMate Duel 375/ha CheckMate Duel 250/ha Insecticide only Mean Cumulative Adult CM +/- SEM /8 5/22 6/5 6/19 7/3 7/177/318/148/289/119/25 C B B AB A No MD Isomate CM/OFM TT CheckMate Duel 500/ha CheckMate Duel 375/ha CheckMate Duel 250/ha Figure 4-5 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult CM in monitoring traps baited with CM 1X and DA Combo lures in Experiment 2. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Tukey s HSD α = 0.05.

98 81 Mean Cumulative OFM +/- SEM /18 5/8 5/28 6/17 7/7 7/27 8/16 9/5 A B* Isomate CM/OFM TT* CheckMate Duel 500/ha* CheckMate Duel 375/ha* CheckMate Duel 250/ha* No MD Figure 4-6 The effect of Isomate CM/OFM TT and three CheckMate Duel dispenser densities on the cumulative capture of adult OFM in monitoring traps baited with OFM lures in 2008 Experiment 2. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = Table 4-5 Percent CM/OFM injured fruit in all treatments in the dispenser density study during farm 1, Experiment 2. Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/13/08 Harvest Isomate CM/OFM TT 0.10 (0.07) 0.00 (0.00) CheckMate Duel 500/ha 0.10 (0.10) 0.00 (0.00) CheckMate Duel 375/ha 0.00 (0.00) 0.00 (0.00) CheckMate Duel 250/ha 0.00 (0.00) 0.00 (0.00) Insecticides only 0.05 (0.05) 0.90 (0.24)

99 82 Table 4-6 Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 farm 2, Experiment 2 Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/13/08 Harvest Isomate CM/OFM TT 0.40 (0.22) 0.70 (0.21) CheckMate Duel 500/ha 0.15 (0.08) 2.15 (0.45) CheckMate Duel 375/ha 0.45 (0.22) 6.10 (1.30) CheckMate Duel 250/ha 0.35 (0.17) 8.65 (0.78) Conventional insecticide only 0.50 (0.17) 4.10 (0.58) Table 4-7 Percent CM/OFM injured fruit in all treatments in the dispenser density study during 2008 farm 3, Experiment 2 Mean number of CM/OFM injured apples per 100 apples (± SEM) Treatment 7/13/08 Harvest Isomate CM/OFM TT 0.60 (0.17) 0.90 (0.25) CheckMate Duel 500/ha 0.05 (0.05) 0.00 (0.00) CheckMate Duel 375/ha 0.05 (0.05) 0.00 (0.00) CheckMate Duel 250/ha 0.00 (0.00) 0.05 (0.05) Insecticides only 0.20 (0.09) 0.50 (0.12) Table 4-8 Total number of CM and OFM live larvae recovered during fruit injury evaluations in all treatment blocks in Experiment /13/08 Harvest Treatment CM OFM CM OFM Isomate CM/OFM TT CheckMate Duel 500/ha CheckMate Duel 375/ha CheckMate Duel 250/ha Insecticides only

100 83 Cumulative Mean Number of CM +/- SEM /2 6/23 7/14 8/4 8/25 9/15 10/6 A AB AB B* B No MD CheckMate Duel 500/ha* CheckMate Duel 425/ha* CheckMate Duel 375/ha CheckMate Duel 250/ha Isomate CM/OFM TT Figure 4-7 The effect of Isomate CM/OFM TT and four CheckMate Duel dispenser densities on the cumulative capture of adult CM in monitoring traps baited with CM DA Combo lures in 2008 Experiment 3. No MD is insecticides only. Treatments followed by the same letters are not significantly different. Treatments followed by an asterisk are followed by the same letter. Tukey s HSD α = 0.05.

101 84 effectively control CM (Epstein et al. 2006) and OFM ( Charlton and Carde 1981; Stelinski et al. 2005b). A possible explanation for these results is that tree size was not standardized within the replicate blocks. In orchards planted with smaller trees, it is likely that growers achieve more thorough coverage with airblast sprayers because the spray cloud does not have to penetrate thick dense canopies, or reach the uppermost fruit near the top of the canopy. The improved spray coverage in smaller tree canopies most likely provides more effective control of CM larvae. Standardizing tree size in large orchard blocks is more likely to balance the effects of both insecticide treatments and mating disruption effects. Also, pre-existing unknown CM populations across different blocks in the orchard possibly played a role in the increased CM capture in the higher dispenser density treatments in this study. It is highly probable that more CM will be caught in blocks where the population was higher the previous year. During 2008, the highest dispenser density (500/ha) of the CheckMate Duel was consistently the most effective treatment at reducing CM capture when compared to the insecticides only and Isomate CM/OFM TT treatments. My findings are consistent with those of Epstein et al. (2006) who showed increased disruption of CM as dispenser density increased using Isomate C Plus and wax drop dispensers. The 2008 results are also consistent with the findings of Charlton and Carde (1981) and Stelinski et al. (2005) who also showed increased disruption of the related OFM as dispenser density increased. In Experiment 2 all MD dispenser densities were effective at reducing CM capture in pheromone traps when compared to insecticides only. This is not surprising given that Epstein et al. (2006) found that even low rates of dispensers substantially

102 85 reduced CM captures in pheromone traps and higher densities reduced CM capture more than lower densities. However, the reduced CM capture in the 250/ha and 375/ha densities of the CheckMate Duel in Experiment 2 may not be sufficient to prevent unacceptable levels of fruit injury at harvest as seen on farm 2 (Table 4-6). Also, the CheckMate Duel dispenser density of 250/ha in Experiment 2 did not achieve a satisfactory rate ( 16%) of CM trap shutdown. The low level of shutdown indicates that the 250/ha density was likely not effective at preventing CM from mating. In the small plot study (Experiment 3), the Isomate CM/OFM TT and CheckMate Duel densities of 425/ha and 500/ha were the only treatments that reduced CM capture below the level observed in the plots treated with insecticides only. The CheckMate Duel densities of 250/ha and 375/ha were not effective at reducing CM capture when compared to the number of CM captured in the insecticides only treatment. The amount of CM pheromone present in the orchard could be affecting the efficacy of the lower dispenser densities of the CheckMate Duel. The initial amount of CM pheromone in the plots treated with the 250/ha density is less than 70 g active ingredient per hectare, which is 42% and 50% of the total amount of pheromone present at 500 dispensers/ha in the Isomate CM/OFM TT and CheckMate Duel plots, respectively. At the 375/ha and 425/ha density rates of the CheckMate Duel, the orchard plots were initially loaded with 75% and 85%, respectively, of the CM active ingredient as the 500/ha density rate. Isomate CM/OFM TT at a density of 500 dispensers/ha contains 36% and 28% more CM pheromone than the 375/ha and 425/ha densities, respectively, of the CheckMate Duel.

103 86 In Experiment 2, the number of live CM larvae found at harvest was high in the CheckMate Duel density of 250/ha and also in the insecticides only treatment (Table 4-8). In Experiment 1 where the CM population was much lower, few larvae were found in all of the treatments. The number of larvae is an indirect measure of MD effectiveness since the MD does not actually kill the larvae. However, since MD is thought to generally work by delaying mating (Vickers 1997), which reduces the fecundity of females (Vickers 1997; Fraser and Trimble 2001; Jones and Aihara-Sasaki 2001; Jones et al. 2008), the number of larvae present in treatments with higher dispenser densities should be lower since the females, if they mate, are probably more likely to mate later in their life than in orchards treated with lower dispenser densities. In Experiments 1 and 2, all dispenser densities of the CheckMate Duel and the Isomate CM/OFM TT treatments plus insecticides effectively reduced OFM capture in pheromone monitoring traps when compared to the insecticides only treatment. In Experiment 1, OFM capture was near zero for the entire season in all MD treatments after the application of dispensers in 2007 and These results are consistent with several other studies where MD was highly effective for disruption of OFM (Atanassov et al. 2002, Il ichev et al. 2002, Roberston and Hull 2002, De Lame et al. 2007). In Experiment 2, OFM capture was again near zero in all treatments except for the CheckMate Duel density of 250/ha where trap shutdown was about 94%. The percent trap shutdown rate for OFM in this treatment was lower when compared to the other treatments where shutdown was 99%. These results are also consistent with the findings of Charlton and Carde (1981) and Stelinksi et al. (2005b) where OFM disruption increased as dispenser density increased. When using effective MD technologies for

104 87 control of OFM, trap shutdown levels are generally very high (> 97%) (De Lame and Gut 2006). My results are also consistent with previous experiments by Stelinski et al. (2007a), Stelinski et al. (2007b), Joshi et al. (2008), Hull et al. (2009) that have shown superior disruption of OFM when compared to that of CM. The lower rate of shutdown in the 250/ha density treatment was due to increased OFM capture in the second half of the season. The 94% shutdown rate would indicate that under high OFM pressure the amount of pheromone being released late in the season by the low dispenser density was not enough to effectively disrupt OFM. The small size of the OFM populations in the other experiments was the likely reason that no differences between MD treatments were found. In Experiment 3 for instance, the OFM population was extremely low (< 2 OFM per trap for the entire season). Release rate analysis of the CheckMate Duel and Isomate CM/OFM TT shows that the CheckMate Duel effectively releases OFM pheromone for about 60 days (Appendix C-3, 4). After 60 days, the release rate stays relatively low for the CheckMate Duel. Whereas, for Isomate CM/OFM TT, the pheromone release rate is more constant for about days. This low release rate from the CheckMate Duel after 60 days can partially explain the failure of the 250/ha treatment in the presence of a high OFM population in Experiment 2. There may not have been enough pheromone being released from the dispensers at the lowest dispenser density after 60 days to disrupt OFM in the presence of high populations. Interestingly, the amount of active ingredient of OFM pheromone within the orchard in the Isomate CM/OFM TT treatment at a density of 500/ha is about 10 g less than in the CheckMate Duel density of 250/ha. However, the Isomate CM/OFM TT

105 88 technology was still effective while the CheckMate Duel density of 250/ha was not as effective in Experiment 2. The initial amount of active ingredient in the CheckMate Duel versus the Isomate CM/OFM TT coupled with the OFM release rate data (Appendix C-3, 4) would indicate that the release rate of OFM pheromone is the more likely reason for the failure of the CheckMate Duel density of 250/ha in Experiment 2. The number of OFM larvae found at harvest was highest in the CheckMate Duel dispenser densities of 250/ha and 375/ha in Experiment 2 (Table 4-8). No OFM larvae were recovered in Experiment 1 during either year. The orchards in Experiment 1 did not have high OFM populations, while the OFM populations in Experiment 2 were high. In Experiment 2, the CheckMate Duel density of 500/ha was the most effective density of this technology at reducing the number of live OFM larvae at harvest. A possible reason that the number of larvae detected at harvest was high is that there was probably not enough pheromone present in the orchard to effectively disrupt OFM. There were no statistical differences in fruit injury during any of the evaluations in Experiment 1. In Experiment 2, fruit injury was highly variable across growers, indicating that MD does not always reduce fruit injury, especially during the first year of use. Fruit injury is again an indirect measure of MD effectiveness as delayed mating should decrease the number of larvae that hatch and cause injury; however, it is likely that the insecticides that were applied had a greater direct effect on fruit injury than the MD since many insecticide products are designed to immediately kill the larvae. Also, the efficacy of certain insecticides and the possibility of resistance need to be accounted for by growers in order to most effectively prevent fruit injury in conjunction with the use of MD.

106 89 The mechanism(s) underlying the increased disruption by higher densities of MD dispensers in these experiments was not clearly discernable due to the nature of the experiments that I performed. In order to properly discern the underlying mechanisms involved, other experiments would be needed to increase the number of point sources releasing pheromone while holding constant the initial amount of pheromone active ingredient across all point source densities (Epstein et al. 2006). Also, flight tunnel studies (Stelinski et al. 2005a) to understand how moths react to the CheckMate Duel dispenser should be performed in conjunction with these other experiments to effectively determine the mechanism(s) behind the disruption of CM and OFM. In conclusion, higher densities of the CheckMate Duel were more effective at reducing adult capture of CM and OFM. Based on the small plot study (Experiment 3), the CheckMate Duel dispenser density of 425/ha in conjunction with supplemental insecticides will likely provide effective control of CM when compared to insecticides only. In orchards where pest populations are high, the CheckMate Duel density of 500/ha should consistently provide the best control of CM and OFM. In orchards where only OFM populations are high, a density of at least 375 dispensers/ha is necessary to shutdown OFM capture in monitoring traps, while the lower density of 250/ha is not effective at reducing capture of OFM under high pest populations. Future research should include a comparison between increasing dispenser density while maintaining the same amount of CM pheromone in the orchard (pheromone loading in individual dispensers decreases as point source density increases) and an adjacent experiment where pheromone loading is increased while keeping the number of dispenser point sources constant. This would allow for a better understanding of whether the increase in the

107 90 amount of pheromone present in the orchard, or the increase in the number of point sources is more important for disruption of CM when using the CheckMate Duel technology.

108 91 Chapter 5 CONCLUSION Mating disruption (MD) is an effective tactic that can be used for the management of codling moth (CM) and oriental fruit moth (OFM) in Pennsylvania. There are many factors that must be understood in order to achieve effective control of these two pests through the use of MD including monitoring trap placement in relation to a dispenser, choice of MD technology, dispenser density, and the duration of time that MD will be used as a management tactic. The placement height of a monitoring trap in relation to MD dispenser placement is very important; the most effective trap placement position for successfully monitoring adult CM and OFM is high in the tree canopy. It is important that the MD dispenser for CM and OFM be placed as high in the tree canopy as possible in order to provide the most effective disruption because the amount of pheromone in the headspace above a dispenser decreases with distance from a dispenser. Placing MD dispensers high will ensure that they are placed above the monitoring traps. Also, the MD dispenser should be placed on a limb that will not bend or break during the season in order to prevent the trap-dispenser relationship from changing throughout the season. If the dispenser location does change over the season, the monitoring trap will not provide an accurate assessment of MD effectiveness because MD does not effectively disrupt the male moth s ability to find monitoring traps that are placed above the dispenser. Future research should address whether there is a difference in moth capture at different heights in relation to a dispenser in tall (i.e., 5 m) apple trees versus shorter

109 92 (i.e., 2-3 m) trees. Canopy structures may affect the pheromone distribution within the tree and the orchard differently in smaller trees in high density plantings where the tree canopies are more open than in taller trees with large canopy volumes in low density plantings. Also, the relationship of trap placement height to multiple dispensers placed at different heights would allow for a better understanding of the most effective placement of dispensers in MD treated orchards. When dispensers are placed at multiple heights within a tree, the pheromone released from the dispensers could possibly be more uniformly distributed throughout the entire canopy and be more likely to effectively shutdown adult captures in monitoring traps placed at any height within the tree canopy. The MD technologies - Isomate CM/OFM TT, Cidetrak OFM, and OFM Disrupt Micro-Flakes were all effective at reducing adult OFM capture in apple orchards. However, the CM Disrupt Micro-Flake treatment was not as effective as the TRE #9940 (an experimental formulation containing both CM pheromone and the kairomone - pear ester) and Isomate CM/OFM TT treatments at reducing adult capture of CM, but the CM Disrupt Micro-Flake treatment plus insecticides was more effective at reducing CM adults captured in monitoring traps than just insecticides alone. All three CM MD treatments reduced CM capture between 74-80% in traps baited with CM DA Combo lures and 61-81% in traps baited with CM 1X lures. Whereas the insecticides only treatment reduced CM capture by only 53% in traps baited with CM DA Combo and 28% in traps baited with CM 1X lures from one year to the next. The increased efficacy of the MD treatments from year to year will likely reduce the amount of insecticides used over time as insect populations are lowered.

110 93 Future research endeavors should focus on concentrating the Micro-Flakes to create pockets that act similar to larger point sources, which will place more pheromone in localized areas over a longer period of time. In addition, an analysis of the release rate of CM pheromone from the Disrupt Micro-Flakes should be conducted to better understand and reveal any possible problems with performance. Also, flight tunnel studies to understand the mechanisms through which the Disrupt Micro-Flakes disrupt CM mating should be conducted. Understanding the mechanisms would help to explain the ineffectiveness of the Disrupt Micro-Flakes at shutting down CM capture in monitoring traps. While both the Cidetrak OFM and TRE #9940 formulations were effective at reducing capture of OFM and CM, respectively, the development of a single Cidetrak dispenser technology for CM and OFM should also be considered. Currently, two different Cidetrak dispensers are necessary to simultaneously disrupt both of these species and the labor to apply two different dispensers at two different densities is time consuming and costly. Higher densities of the CheckMate Duel, 425 and 500 dispensers/ha, were more effective at reducing adult capture of CM and OFM than lower densities. Based on the small plot study, the CheckMate Duel dispenser density of 425/ha in conjunction with insecticides should effectively reduce CM capture in monitoring traps when compared to an insecticides only treatment. In orchards where pest populations are high, the CheckMate Duel density of 500/ha should consistently provide the best control of CM and OFM. In orchards where only OFM populations are high, a density of at least 375 dispensers/ha is likely necessary to shutdown OFM capture in monitoring traps since lower densities were not effective. Isomate CM/OFM TT at 500 dispensers/ha along with

111 94 a supplemental insecticide program was effective at reducing adult CM and OFM captures in monitoring traps under all conditions. Future research should include a comparison between increasing dispenser density while maintaining the same amount of CM pheromone in the orchard (i.e., pheromone loading in individual dispensers decreases as point source density increases) and an adjacent experiment where pheromone loading rate is increased while keeping the number of dispenser point sources constant. This would allow for a better understanding of whether the increase in the amount of pheromone present in the orchard, or the increase in the number of point sources is more important for disruption of CM when using the CheckMate Duel technology.

112 95 LITERATURE CITED Ahmad, T.R. and Al-Gharbawi, Z.A Effects of pheromone trap design and placement on catches of codling moth males. J. Appl. Entomol. 102: Angeli, G., Anfora, G., Baldessari, M., Germinara, G.S., Rama, F., De Cristofaro, A., Ioriatti, C Mating disruption of codling moth Cydia pomonella with high densities of Ecodian sex pheromone dispensers. J. Appl. Entomol. 131: Atanassov, A., Shearer, P.W., Hamilton, G., Polk, D Development and implementation of a reduced risk peach arthropod management program in New Jersey. J. Econ. Entomol. 95: Atterholt, C.A Controlled release of insect sex pheromones from sprayable, biodegradable materials for mating disruption. PhD dissertation, University of California at Davis, Davis, CA. Baker, T.C., Fadamiro, H.Y., Cosse, A.A Moth uses fine tuning for odor resolution. Nature. 393: 530. Baker, T.C., and Heath J.J Pheromones: Function and use in insect control. PP In: Gilbert, L.I, Iatrou, K. and Gill, S.S. (eds.), A Comprehensive Molecular Insect Science 1 st ed. Vol. 6. Elsevier, Boston. Barnes, M.M Tortricids in Pome and Stone Fruits. pp In: L. P. S. Van Der Geest and H. H. Evenhuis (eds.), Tortricid Pests. Elsevier, Amsterdam. Barrett, B.A Male codling moth (Lepidoptera: Tortricidae) in pheromone and virgin female moth-baited traps at different tree heights in small orchard blocks. Environ. Entomol. 24: Bartell, R.J Mechanisms of communication disruption by pheromone in the control of lepidoptera: a review. Phys. Entomol. 7: Berger, R.G., Drawert, F., Schrafstetter, B Natural occurrence of Octenoic-, Decenoic-, and Decadienoic acid ethyl esters in red delicious apples. Z Lebensm Unters Forsch178: Beroza, M., Gentry, C.R., Blythe, J.L., and Muschik, G.M Isomer content and other factors influencing captures of oriental fruit moth by synthetic pheromone traps. J. Econ. Entomol. 66: Borden, A.B Some field observations on codling moth behavior. J. Econ. Entomol. 22:

113 96 Brown, D.J., and Il ichev, A.L The potential for the removal of organo-phosphate insecticides from the stone-fruit production in the Goulburn Valley, Australia. Acta Hort. 525: Brunner, J., Welter, S., Calkins, C, Hilton, R., Beers, E., Dunley, J., Unruh, T., Knight, A., Van Steenwyk, R., and Buskirk, P. Van Mating disruption of codling moth: a perspective from the western United States. IOBC wprs Bulletin. 25. Calkins, C.O Review of the codling moth areawide suppression program in the western United States. J. Agric. Entomol. 15: Calkins, C.O. and Faust, R.J Overview of areawide Programs and the program for suppression of codling moth in the western USA directed by the United States Department of Agriculture Agricultural Research Service. Pest Manag. Sci. 59: Carde, A.M., Baker, T.C., and Carde, R.T Identification of a four-compenent sex pheromone of the female oriental fruit moth, Grapholitha molesta (Lepidoptera: Tortricidae). J. Chem. Ecol. 5: Carde, R.T., Baker, T.C., and Castrovillo, P.J Disruption of sexual communication in Laspeyresia pomonella (codling moth), Grapholitha molesta (oriental fruit moth) and G. prunivora (lesser appleworm) with hollow fiber attractant sources. Entomol. Exp. Appl. 22: Carde, R.T., and Haynes, K.F Structure of the pheromone communication channel in moths. PP In: Carde, R.T, and Millar, J.G. (eds.), Advances in Insect Chemical Ecology, Cambridge University Press, Boston. Caro, J.H., Glotfelty, D.E. and Freeman, H.P (Z)-9-Tetradecen-1-ol formate distribution and Dissipation in the Air within a corn crop after emission from a controlled-release formulation. J. Chem. Ecol. 6: Charlton, R.E., and Carde, R.T Comparing the effectiveness of sexual communication disruption in the oriental fruit moth (Grapholitha molesta) 1 using different combinations and dosages of its pheromone blend 2. J. Chem. Ecol. 3: De Lame, F.M., and Gut, L.J Effect of monitoring trap and mating disruption dispenser application heights on captures of male Grapholita molesta (Busck; Lepidoptera: Tortricidae) in pheromone and virgin female-baited traps. Environ. Entomol. 35:

114 97 De Lame, F., Miller, J.R., Atterholt, C.A., and Gut, L.J Development and evaluation of an emulsified paraffin wax dispenser for season-long mating disruption of Grapholita molesta in commercial peach orchards. J. Econ. Entomol. 100: Epstein, D., McGhee, P., and Gut, L Making codling moth mating disruption work in Michigan: Adopting an area-wide approach to managing codling moth in Michigan apple production. Michigan State University Fruit CAT Newsletter. 20. Epstein, D.L., Stelinski L.L., Reed, T.P., Miller, J.R., and Gut, L.J Higher densities of distributed pheromone sources provide disruption of codling moth (Lepidoptera: Tortricidae) superior to that of lower densities of clumped sources. J. Econ. Entomol. 99: Evenden, M.L. and McLaughlin, J.R Factors influencing the effectiveness of an attracticide formulation against the oriental fruit moth, Grapholita molesta. Entomol. Exp. Appl. 112: Evenden, M.L. and McLaughlin, J.R Male oriental fruit moth response to a combined pheromone-based attracticide formulation targeting both oriental fruit moth and codling moth (Lepidoptera: Tortricidae). J. Econ. Entomol. 98: Fernandez, D., Bosch, D., Cichon, L., Avilla, J A secondary sexual character for sex determination of Cydia pomonella (L.) (Lepidoptera: Tortricidae) adults, trapped with kairomone lures. IOBC wprs Bulletin. 30. Fraser, H.W. and Trimble, R.M Effect of delayed mating on reproductive biology of the oriental fruit moth (Lepidoptera: Tortricidae). Can. Entomol. 133: Gentry, C.R., Beroza, M., Blythe, J.L. and Bierl, B.A Efficacy trials with the pheromone of the oriental fruit moth and data on the lesser appleworm. J. Econ. Entomol. 67: Gut, L.J., and Brunner, J.F Pheromone-based management of codling moth (Lepidoptera: Tortricidae) in Washington apple orchards. J. Agric. Entomol. 15: Hern, A., and Dorn, S Sexual dimorphism in the olfactory orientation of adult Cydia pomonella in response to α-farnesene. Entomol. Exp. Appl. 92: Howell, J.F., Schmidt, R.S., Horton, D.R., Khattak, S.U.K., and White, L.D Codling moth: male moth activity in response to pheromone lures and pheromone-baited traps at different elevations within and between trees. Environ. Entomol. 19:

115 98 Hull, L.A., Krawczyk, G., Bohnenblust, E., and Biddinger, D Expansion of an area-wide pheromone mating disruption approach to control two major fruit pests in Pennsylvania orchards. Penn Fruit News 87: Hull, L., Krawczyk, G., Bohnenblust, E., Biddinger, D Expansion of an area-wide pheromone mating disruption approach to control two major fruit pests in Pennsylvania orchards year 2. Penn Fruit News. 87: Hull, L., Krawczyk, G., Bohnenblust, E., Zaman, F., and Biddinger, D Expansion of an area-wide pheromone mating disruption approach to control two major fruit pests in Pennsylvania orchards year 3. Penn Fruit News 89: Hull, L.A., Krawczyk, G., Ellis, N Management tactics for the oriental fruit moth (Grapholita molesta) in Pennsylvania apple orchards. Penn Fruit News. 81: Il ichev, A.L., Williams, D.G., and Gut, L.J Dual pheromone dispenser for combined control of codling moth Cydia pomonella (L.) and oriental fruit moth Grapholita molesta (Busck) (Lep., Tortricidae) in pears. J. Appl. Entomol. 131: Il ichev, A.L., Gut, L.J., Williams, D.G., Hossain, M.S., and Jerie, P.H Areawide approach for improved control of oriental fruit moth Grapholita molesta (Busck) (Lepidoptera: Tortricidae) by mating disruption. Gen. Appl. Entomol. 31: Il ichev, A.L., Williams, D.G. and Milner, A.D Mating disruption barriers in pome fruit for improved control of oriental fruit moth Grapholita molesta Busck (Lep., Tortricidae) in stone fruit under mating disruption. J. Appl. Entomol. 128: Jones, V.P. and Aihara-Sasaki, M Demographics analysis of delayed mating in mating disruption: a case study with Cryptophelbia illepida (Lepidoptera: Tortricidae). J. Econ. Entomol. 94: Jones, V.P., Wiman, N.G., and Brunner, J.F Comparison of delayed female mating on reproductive biology of codling moth and obliquebanded leafroller. Environ. Entomol. 37: Joshi, N.K. and Hull, L.A Efficiency of different lure types for monitoring codling moth adults in apple orchards a research update. Penn Fruit News 89: Joshi, N.K, Hull, L.A., Krawczyk, G., Rajotte, E.G Field results of mating disruption technologies for the control of codling moth, Cydia pomonella (L.),

116 99 and oriental fruit moth, Grapholita molesta (Busck) in Pennsylvania apple orchards. Aspects Appl. Biology. 84: Karg, G. and Sauer, A.E Spatial distribution of pheromone in vineyards treated for mating disruption of the grape vine moth Lobesia botrana measured with electroantennograms. J. Chem. Ecol. 21: Knight, A.L The impact of codling moth (Lepidoptera: Tortricidae) mating disruption on apple pest management in Yakima Valley, Washington. J. Entomol. Soc. Brit. Columbia. 92: Knight, A.L Monitoring codling moth (Lepidoptera:Tortricidae) with passive interception traps in sex pheromone-treated apple orchards. J. Econ. Entomol. 93: Knight, A.L Using the kairomone lure DA2313 to monitor codling moth in apple and pear. Proceedings, Western Orchard Pest and Disease Management Conference, 9-11 January Portland, OR. Knight, A.L Assessing the mating status of female codling moth (Lepidoptera: Tortricidae) in orchards treated with sex pheromone traps baited with Ethyl (E,Z)- 2,4-decadienoate. Environ. Entomol. 35: Knight, A.L Influence of within-orchard trap placement on catch of codling moth (Lepidoptera: Tortricidae) in sex pheromone-treated orchards. Environ. Entomol. 36: Knight, A.L., Croft, B.A., and Bloem, K.A Effect of mating disruption dispenser placement on trap performance for monitoring codling moth (Lepidoptera: Tortricidae). J. Entomol. Soc. Brit. Columbia. 96: Knight, A.L., Hilton, R., Light, D.M Monitoring codling moth (Lepidoptera: Tortricidae) in apple with blends of ethyl (E,Z)-2,4-decadienoate and codlemone. Environ. Entomol. 34: Knight, A.L., and Larsen, T.E Improved deposition and performance of a microencapsulated sex pheromone formulation for codling moth (Lepidoptera: Tortricidae). J. Entomol. Soc. Brit. Columbia. 101: Knight, A.L., and Light, D.M Use of (E,Z)-2-4 decadienoic acid in codling moth management. 2. Stimulation of oviposition. J. Entomol. Soc. Brit. Columbia. 101:

117 100 Knight, A.L., and Light, D.M. 2005a. Seasonal flight patterns of codling moth (Lepidoptera: Tortricidae) monitored with pear ester and codlemone-baited traps in sex pheromone-treated apple orchards. Environ. Entomol. 34: Knight, A.L. and Light, D.M. 2005b. Developing action thresholds for codling moth (Lepidoptera: Tortricidae) with pear ester- and codlemone-baited traps in apple orchards treated with sex pheromone mating disruption. Can. Entomol. 137: Koch U.T. and Witzgall, P Pheromones for insect control in orchards and vineyards. IOBC\WPRS Bulletin. 24: Krawczyk, G. and Hull, L.A Factors in monitoring and controlling codling moth populations in PA apple orchards. Penn Fruit News. 85: Krawczyk, G Monitoring insecticide resistance in field populations of oriental fruit moth and codling moth collected from Pennsylvania fruit orchards. Penn Fruit News. 86: Light, D.M., Knight, A.L., Henrick, C.A., Rajapaska, D., Lingren, B., Dickens, J.C., Reynolds, K.M., Buttery, R.G., Merrill, G., Roitman, J., Campbell, B.C A pear derived kairomone with pheromonal potency that attracts male and female codling moth, Cydia pomonella (L.). Naturwissenschaften. 88: Lo, P.L., and Cole, L.M Impacts of pheromone mating disruption and pesticides on oriental fruit moth (Grapholita molesta) on peaches. New Zealand Plant Prot. 60: Madsen, H.F., and Vakenti, J.M Codling moths: female baited and synthetic pheromone traps as population indicators. Environ. Entomol. 1: Mattheis, J.P., Fellman, J.K., Chen, P.M., Patterson, M.E Changes in the headspace volatiles during physiological development of Bisbee delicious apple fruit. J. Agric. Food Chem. 39: McNally, P.S. and Barnes, M.M Effects of codling moth pheromone trap placement, orientation and density on trap catches. Environ. Entomol. 10: Miller, J.R., Gut, L.J., de Lame, F.M., Stelinski, L.L. 2006a. Differentiation of competitive vs. non-competitive mechanisms mediating disruption of moth sexual communication by point sources of sex pheromone (Part 1): Theory. J. Chem. Ecol. 32: Miller, J.R., Gut, L.J., de Lame, F.M., Stelinski, L.L. 2006b. Differentiation of competitive vs. non-competitive mechanisms mediating disruption of moth sexual

118 101 communication by point sources of sex pheromone (Part 2): case studies. J. Chem. Ecol. 32: Milli, R., Koch, U.T. and de Kramer, J.J EAG measurements of pheromone distribution in apple orchards treated for mating disruption of Cydia pomonella. Entomol. Exp. Appl. 82: Minitab Inc Statistical Software Version 14. Myers, C.T., Hull, L., Krawczyk, G Comparative survival rates of oriental fruit moth (Lepidoptera:tortricidae) larvae on shoots and fruit of apple and peach. J. of Econ. Entomol. 99: Pfeiffer, D.G, Kaakeh, W., Killian, J.C., Lachance, M.W., and Kirsch, P Mating disruption for control of damage by codling moth in Virginia apple orchards. Entomol. Exp. Appl. 67: Riedl, H The importance of pheromone trap density and trap maintenance for the development of standardized monitoring procedures for the codling moth (Lepidoptera: Tortricidae). Can. Entomol. 112: Riedl, H., Croft, B.A., and Howitt, A.J Forecasting codling moth phenology based on pheromone trap catches and physiological-time models. Can. Entomol. 108: Riedl, H., Howing, S.A., Barnett, W.W., and DeTar, J.E Relationship of withintree placement of the pheromone trap to codling moth catches. Environ. Entomol. 8: Robertson, S., Hull, L Areawide mating disruption of the oriental fruit moth, Grapholita molesta, in Pennsylvania apples and peaches Penn Fruit News. 82: Roelofs, W Sex pheromone of the oriental fruit moth. Nature. 224: 723. Roelofs, W., Comeau, A., Hill, A., Milicevic, G Sex attractant of the codling moth: characterization with electroantennogram technique. Science. 174: Rothschild, G.H.L Control of oriental fruit moth (Cydia molesta (Busck) (Lepidoptera, Tortricidae)) with synthetic female pheromone. Bull. Entomol. Res. 65:

119 102 Rothschild G.H.L., and Minks, A.K Some factors influencing the performance of pheromone traps for the oriental fruit moth in Australia. Entomol. Exp. Appl. 22: Rothschild, G.H.L., and Vickers, R.A Biology, ecology and control of the oriental fruit moth. PP In: L. P. S. Van Der Geest and H. H. Evenhuis (eds.), Tortricid Pests. Elsevier, Amsterdam Stelinski, L.L., Gut, L.J., Ketner, K.C., and Miller, J.R. 2005a. Orientational disruption of codling moth, Cydia pomonella (L.) (Lep., Tortricidae), by concentrated formulations of microencapsulated pheromone in flight tunnel assays. J. Appl. Entomol. 129: Stelinski, L.L., Gut, L.J., Mallinger, R.E., Epstein, D., Reed, T.P., and Miller, J.R. 2005b. Small plot trials documenting effective mating disruption of oriental fruit moth by using high densities of wax-drop pheromone dispensers. J. Econ. Entomol. 98: Stelinski, L.L., Gut, L.J., Haas, M., McGhee, P., Epstein, D. 2007a. Evaluation of aerosol devices for simultaneous disruption of sex pheromone communication in Cydia pomonella and Grapholita molesta (Lepidoptera: Tortricidae). J. Pest Sci. 80: Stelinski, L.L., McGhee, P., Haas, M., Il ichev, A.L., Gut, L.J. 2007b. Sprayable microencapsulated sex pheromone formulations for mating disruption of four tortricid species: effects of application height, rate, frequency and sticker adjuvant. J. Econ. Entomol. 100: Stelinski, L.L., Il Ichev, A.L., and Gut, L.J Efficacy and release rate of reservoir pheromone dispensers for simultaneous mating disruption of codling moth and oriental fruit moth (Lepidoptera: Tortricidae). J. Econ. Entomol. 102: Stelinski, L.L., McGhee, P., Grieshop, M., Brunner, J., and Gut, L.J Efficacy and mode of action of female-equivalent dispensers of pheromone mating disruption of codling moth. Agric. and Forest Entomol. 10: Stoeckli, A., Mody, K., and Dorn, S Influence of canopy aspect and height on codling moth (Lepidoptera: Tortricidae) larval infestation in apple, and relationship between infestation and fruit size. J. Econ. Entomol. 101: Suckling, D.M., Green, S.R., Gibb, A.R., and Karg, G. 1999a. Predicting atmospheric concentration of pheromone in treated apple orchards. J. of Chem. Ecol. 25:

120 103 Suckling, D.M., Green, S.R., Gibb, A.R., Karg, G. 1999b. Atmospheric concentrations affect the behavior of lightbrown apple moth (Lepidoptera: Tortricidae) in the orchard. J. Chem. Ecol. 25: Sutherland, O.R.W., Wearing, C.H., and Hutchins, R.F.N Production of α- farnesene, an attractant and oviposition stimulant for codling moth by developing fruit of ten varieties of apple. J. Chem. Ecol. 3: Tasin, M., Demaria, D., Ryne, C., Cesano, A., Galliano, A., Anfora, G., Ioratti, C., and Alma, A Effect of anti-hail nets on Cydia pomonella behavior in apple orchards. Entomol. Exp. Appl. 129: Thwaite, W.G., and Madsen, H.F The influence of trap density, trap height, outside traps and trap design on Cydia pomonella (L.) captures with sex pheromone traps in New South Wales apple orchards. J. Aust. Entomol. Soc. 22: Tomaszewska, E., Herbert, V.R., Brunner, J.F., Jones, V.P., Doerr, M., and Hilton, R Evaluation of pheromone release from commericial mating disruption dispensers. J. Agric. Food Chem. 53: Trimble, R.M., Pree, D.J., Vickers, P.M. and Ker, K.W Potential of mating disruption using sex pheromone for controlling the grape berry moth, Endopizea viteana (Clemens) (Lepidoptera: Tortricicdae, in Niagara peninsula, Ontario vineyards. Can. Entomol. 123: Vickers, N.J Winging it: Moth flight behavior and responses of olfactory neurons are shaped by pheromone plume dynamics. Chem. Senses 31: Vickers, R.A., and Rothschild, G.H.L Use of sex pheromones for control of codling moth. PP In: L. P. S. Van Der Geest and H. H. Evenhuis (eds.), Tortricid Pests. Elsevier, Amsterdam. Vickers, R.A., Thwaite, W.G., Williams, D.G., and Nicholas, A.H Control of codling moth in small plots by mating disruption: along and with limited insecticide. Entomol. Exp. Appl. 86: Vickers, R.A Effect of delayed mating on oviposition pattern, fecundity and fertility in codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae). Aust. J. of Entomol. 36: Weissling, T.J. and Knight, A.L Vertical distribution of codling moth adults in pheromone-treated and untreated plots. Entomol. Exp. Appl. 77:

121 Witzgall, P., Stelinski, L., Gut, L. Thomson, D Codling moth management and chemical ecology. Annu. Rev. Entomol. 53:

122 105 Appendix A Maps of Treatment and Trap Locations Figure A-1 Map of treatment layout for MD technology comparison study in 2007 and The second insecticide only block for 2008 is not shown. Treatments are: Insecticides only (solid black line), Disrupt Micro-Flakes (blue fill), TRE # 9940 and Cidetrak OFM (yellow long dash), Isomate CM/OFM TT (red short dash). Numbers correspond to replicates.

123 106 Figure A-2 Map of treatment layout in the CheckMate Duel dispenser study in Insecticides only blocks are not shown. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2).

124 107 Figure A-3 Map of treatment layout in the CheckMate Duel dispenser study in Insecticide only blocks are not shown. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2).

125 108 Figure A-4 Map of treatment layout in the CheckMate Duel dispenser study on the farm near Bendersville, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) and Insecticides only (CV).

126 109 Figure A-5 Map of treatment layout in the CheckMate Duel dispenser study on the farm near Gardners, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3), CheckMate Duel 500/ha (T2) and insecticides only (CV).

127 110 Figure A-6 Map of treatment layout in the CheckMate Duel dispenser study on the farm near Peach Glen, PA in Experiment 2. Treatments are: Isomate CM/OFM TT (T1), CheckMate Duel 250 dispensers/ha (T4), CheckMate Duel 375/ha (T3),CheckMate Duel 500/ha (T2) and insecticides only (CV).

128 111 Figure A-7 Example of trap locations within each treatment block for the MD technology comparison and CheckMate Duel dispenser density study Experiments 1 and 2 for both 2007 and 2008.