Trap Design and Bait Longevity Experiments for Managing Spotted Wing Drosophila

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Trap Design and Bait Longevity Experiments for Managing Spotted Wing Drosophila Chris Castro, Rebekah Jackson, Ashlynn Johnson, Randy Lee, and Mark Salser All are members of Snohomish County Fruit Society, a chapter of the Western Cascade Fruit Society ABSTRACT Our goal was to identify effective and practical trap design and bait practices for backyard fruit growers for use in monitoring and potentially mass trapping of spotted wing Drosophila (SWD). In the Trap Design experiment we collected and counted the SWD catch weekly in five types of traps placed in blueberries and strawberries over an eight week period. In the Bait Longevity experiment we used a single trap design and collected SWD catch according to five treatments (filtered weekly or changed every one, two, three, or four weeks) over an eight week period. In the Trap Design experiment, the water bottle with holes and the spice bottle with holes caught more SWD than other designs; the deli cup caught fewer SWD than other designs. In the Bait Longevity experiment, number of SWD caught per week dropped off progressively as the length between changing the bait increased. For backyard fruit growers, our study suggests a water bottle or spice bottle with holes, and with weekly changing of the bait, allows the most effective trapping of SWD. INTRODUCTION The spotted wing Drosophila (SWD), an invasive fly species with a rapidly expanding range, was first seen along the I- corridor of Washington in 009 and has quickly moved across the state since then. While most vinegar flies prefer rotting fruit that has fallen from the plant, SWD prefer ripening to overripe fruit that is still hanging on the plant. Infested fruit results in larvae feeding inside fruit which quickly begins to deteriorate and becomes a target for secondary pests as well as fungal and bacterial infections. According to Dave Pehling, Snohomish County Extension Agent who specializes in Community Horticulture Pest Management, SWD is endemic in Snohomish County. It has been shown that early intervention made possible by monitoring with traps reduces the overall damage by SWD. Use of traps as an effective control through mass trapping is currently used in China which has been dealing with SWD since the early 900's. Providing home orchardists with a SWD trapping option that is effective, affordable, durable, and easy to use could encourage SWD control. In the present study, we compared the effectiveness of different trap designs for SWD trapping. Trapping is challenging for home orchardists due to the time required to empty and rebait traps, which has typically been done weekly in prior studies. We assessed whether bait can be changed less than weekly without compromising trap effectiveness. METHODS Setting: This study was conducted over an 8-week period at Skipley Farms in Snohomish County, WA. All traps were placed among strawberries or blueberries (Figure ). The blueberries were all early to middle season varieties but fairly young plants so fruit was limited. The strawberries were ever-bearing and intercropped with grapes in alternating rows. For deploying traps, inch wooden stakes were driven 0- inches into the ground, with a screw sticking out at the top for hanging the traps. Blueberries Figure : Trap placement crop locations. Strawberries (and Grape Intercrop)

The study consisted of two separate experiments: the Trap Design experiment and the Bait Longevity experiment. Trap Design experiment: We compared five trap designs: a 6-ounce water bottle with holes, a 6-ounce spice bottle with holes (/6th inch diameter), a 6-ounce spice bottle with a in mesh window, a ounce plastic drinking cup with holes, and a -ounce deli container with holes (Figure ). A similar deli container trap design has been used in previous studies of SWD trapping. The experiment consisted of six traps of each design. Traps were grouped into blocks, where each block contained one of each trap type ( traps per block). Three blocks were placed in strawberries, and three blocks were placed in blueberries (Figure ). Each block was placed a minimum of 0 feet from all other blocks. Within each block, stakes for traps were spaced feet apart. At the time of trap placement, traps were filled with 0 ml of bait (apple cider vinegar mixed with liquid dishsoap, ml of soap per.78 L of vinegar). Each week, all bait and trapped insects were removed from each trap into an individually labeled whirlpack for insect identification and counting. Traps were re-filled with 0 ml of bait. The traps were then randomly moved to a different stake within the same block, to ensure that any spatial variation in SWD density within a block would not be correlated with trap design. Using a microscope, study personnel counted the number of male SWD and female SWD captured in each trap each week. They also counted the number of non-spotted wing Drosophila and then number of non-drosophila insects. All study personnel involved in counting SWD were trained in SWD identification by a professional entomologist and had access to a SWD dichotomous key developed by the Oregon Department of Agriculture. Any questionable identifications were verified by additional team members and/or Snohomish County Extenstion agents. 6 oz. Water Bottle w/ Holes 6 oz. Spice Bottle w/ 6 oz. Spice Bottle w/ Holes Mesh oz. Plastic Drink Cup w/ Holes oz. Deli Container w/ Mesh Figure : Trap designs. Trap Design: Blueberry Blocks Trap Design: Strawberry Blocks Figure : Trap Design block arrangements. =6 oz water bottle w/holes =6 oz. spice bottle w/ holes =6 oz. spice bottle w/ mesh = oz. plastic drink cup w/ holes = oz. deli container w mesh

Bait Longevity experiment: We used 6 pz. spice bottles with holes for all traps in the Bait Longevity experiment. We tested five bait changing options: changing the bait weekly changing the bait every two weeks, changing the bait every three weeks, changing the bait every four weeks, and filtering insects weekly but leaving the same bait in the trap. The experiment consisted of three traps of each bait option. The traps were grouped into three blocks, where each block contained one trap of each bait option (Figure ). The blocks were all placed in blueberries. As with the Trap Design experiment, within each block, stakes for traps were spaced five feet apart, and each block was a minimum of 0 feet from other blocks. At trap placement, traps were filled with 0 ml of bait. For the bait changing options, bait and trapped insects were removed as scheduled to an individually labeled whirlpack for insect identification and counting. Traps were then re-filled with 0 ml of bait, and traps were moved randomly to a different stake within the same block. For the weekly filtration traps, study personnel used a strainer to remove all insects from the trap and returned the bait to the trap. SWD were then counted as in the Trap Design experiment. During the experiment, Skipley Farms had low blueberry yield due to young blueberry plants and a mummyberry infestation. Because of this, during the last four weeks of the study we added additional traps, three of changing bait weekly and three of changing bait every four weeks. These traps were placed in three blocks (one trap of each type per block) among strawberries, with the same spacing as the other traps. This was done only to confirm our findings in the blueberries. =changed weekly =filtered weekly but not changed =changed ever weeks, no filtering =changed ever weeks, no filtering =changed ever weeks, no filtering Bait Longevity: Blueberry Blocks Bait Longevity: Strawberry Blocks Figure : Bait Longevity block arrangements. Analysis: SWD counts were calculated as SWD per trap per week. We first tested whether fruit type (strawberries or blueberries), block location in the field, trap location within each block, or collection week were associated with SWD per trap-week using analysis of variance (ANOVA). We then fit Poisson regression models for each experiment, with SWD per trap-week as the dependent variable. In the Trap Design experiment, we included each trap type as a predictor, and for the Bait Longevity experiment, we included each bait changing option as a predictor. In both analyses we included any variables found to be associated with SWD counts in the ANOVA models. RESULTS In total, the project caught,7 spotted wing Drosophila (SWD) over the eight weeks of the project, with a mean of 6. SWD (range, 0 to 8) caught per trap-week. The project also caught,89 Drosophila of other species (mean. per trap-week), and,07 non-fruit fly insects. The yield of 8 SWD in a single trapweek (a water bottle in strawberries in the Trap Design experiment) was an extreme outlier, as no other traps caught more than 0 SWD per week. All analyses presented below exclude this outlier. However, the conclusions are unchanged if this outlier is included.

Based on ANOVA analysis, significantly more SWD were caught by traps placed in strawberries (mean =. SWD per trap-week) than traps in blueberries (mean =. SWD per trap-week). SWD trapped also varied significantly with study week, with weeks,,, and 7 having the highest trap yield (Figure 6). Figure 6: SWD per trap per week, by collection week. In contrast, SWD per trap-week did not vary significantly between the block locations or between the trap locations within each block. We fit a Poisson regression model to the Trap Design experiment data, including trap type, collection week, and fruit type as predictors, using the spice bottle with holes as a reference group. The water bottle tended to catch % more SWD per trap-week than did the spice bottle with holes. This difference was statistically significant, meaning it was unlikely to be due to chance. The other trap designs were inferior to the spice bottle with holes, collecting % fewer SWD (deli cup), 7% fewer SWD (drink cup), or % fewer SWD (spice bottle with mesh). These differences were also unlikely to be due to chance. To put these differences in context, the table below shows the number of SWD we would expect each trap to catch in a high collection week (week among the strawberries) and a low collection week (week 6 among the blueberries) (Table ). Trap High collection week Spice bottle with holes 6 Water bottle 0 Deli cup 0 Drink cup 9 Spice bottle with mesh 8 Table : Expected number of SWD trapped per week Low collection week Finally, we fit a Poisson regression model to the Bait Longevity experiment data, including bait changing option, collection week, and fruit type as predictors, using weekly changing as the reference group. Compared with changing the bait weekly, changing the bait every two weeks resulted in 6% fewer SWD per trap-week. Compared with changing the bait weekly, changing the bait either every three weeks resulted in 7% fewer SWD per trap-week, while changing the bait every four weeks resulted in 8% fewer SWD per

trap-week. Filtering the bait every week resulted in 88% fewer SWD per trap-week. All these differences were unlikely to be due to chance. DISCUSSION A limitation of the study was the reduced crop in the blueberries which greatly reduced the numbers of attractants to the flies in that area. Though we did not test for this directly, the farm had implemented improved sanitation practices starting in week two of our study. They were suffering high losses in their strawberries and there was an apparent decrease in flies caught though the strawberries were still in the height of ripening. This might suggest that sanitation practices have the potential to impact SWD populations and fruit damage. There are many interesting questions that were raised during the course of this study and that we would suggest as future studies. In order to determine the potential of mass trapping, trap density should be evaluated. In addition, trap location particularly at what height the trap is hung and at what season during the year could greatly affect trap effectiveness. SWD trapping for home orchardists needs to be effective, affordable, and easy to use. In this experiment, we tested various trap designs and bait changing options that could be used in the home orchard context. Our first important finding is that changing the bait less than weekly resulted in dramatic reductions in trapping effectiveness. Changing the bait every two weeks cut trapping effectiveness in half compared with changing bait weekly, and longer interval were even less effective. Filtering the bait was also ineffective. Thus, home orchardists hoping to trap SWD for monitoring or control will need to plan on changing trap bait at least weekly. We also found that trapping effectiveness varied with trap design. The most effective design was the 0ounce water bottle with holes, while the plastic water cup and the spice bottle with mesh were least effective. We note that while the water bottle with holes was somewhat more effective than the spice bottle with holes, study personnel found the spice bottles to be much easier to work with. The spice bottles are more durable, easier to handle, and much less prone to leaking bait out the holes when removing bait and trapped insects. Thus, home orchardists may prefer using the spice bottles with holes to the water bottles with holes, even though the water bottles were somewhat more effective for trapping.

Appendices From left to right, top to bottom: -The research team meets to plan the study. -Taking an initial look under the microscope. -Deploying the traps by filling them with Apple Cider Vinegar with a drop of soap. Ready to deploy, July, 0. -Mark lays out the blocks of traps in the strawberries. 6-Our first samples! Along with our collection tubs and data sheets. 7-Labeled samples ready to count. 8-Practicing identification as a team before we begin our weekly counts. 9-Ready to count those flies!

Map of Skipley Farms Research Location Blueberries **Map is not to scale and block location is approximate** Strawberries / Grapes Skipley Farm 78 Skipley Road Snohomish, WA 9890 Gil Schieber, owner = Trap Design blocks = Bait Longevity blocks