Impacts of the Brown Marmorated Stink Bug on California Agriculture. Ryan M. Fernandez. Department of Environmental Studies

Transcription

1 Impacts of the Brown Marmorated Stink Bug on California Agriculture Ryan M. Fernandez Department of Environmental Studies Environmental Policy Thesis California State University, Sacramento December 17 th, 2015

2 Acknowledgments This thesis was conducted in collaboration with California Department of Food and Agriculture, and supported by a grant received by Dr. Charles H. Pickett. I d like to thank the following people for their assistance they had with the study: Dr. Charlie H. Pickett, CDFA Pest Detection Emergency Projects/Biological Control Marypat Stadtherr, CDFA Pest Detection Emergency Projects/Biological Control Viola Popescu, CDFA Pest Detection Emergency Projects Joan Scheiman, CDFA Pest Detection Emergency Projects

3 Abstract: Brown marmorated stink bugs are an invasive urban and agricultural pest that are new to California. Since its introduction to California, the brown marmorated stink bug has been able to establish reproducing populations in six different counties and have been intercepted in over twenty counties state wide. On top of being an urban nuisance these insects have the potential to cause millions of dollars of damage to agriculture based on reports in the Mid-Atlantic States. Currently this insect has not spread to any agricultural areas in California and no major damage has been reported. The goal of this project is to determine if olives, one of California s many specialty crops, are at risk for damage of the brown marmorated stink bug. Commercially grown olives in the United States are only grown in California and it is unknown if they will be affected once the brown marmorated stink bug spreads. Four, two week trials with varying treatments, were conducted to determine if stink bugs are able to survive and reproduce on olives. Insects were able to survive and reproduce on mature olives better than younger olives. However, the insects had higher survival and reproduction rate when a water source was present regardless of the maturity level of olives. Based on the results of this experiment, olives are one of the many specialty crops that are at high risk of what have been damaged by brown marmorated stink bugs in other states. Pest management and pesticide application strategies will have to be developed for olive growers to prevent negative economic consequences to their growing industry from these insects.

4 Table of Contents Page Introduction...1 -California Agriculture California Olive Industry...4 -Project Objectives...5 Methods...6 -Treatment Descriptions...7 -Trial Descriptions Weight Comparison and Damage guidelines Behavior and Feeding Habit guidelines Reproduction, Relative Humidity, and Temperature guidelines Results Control #1: Olives...12 Weight Comparison...12 Relative Humidity/Temperature...12 Olive Observations...13 Survivorship Control #2: Olives + Water...14 Weight Comparison...14 Relative Humidity/Temperature Olive Observations Survivorship Olives + Insects Weight Comparison Egg masses, Egg, Relative Humidity/Temperature Olive Observations Survivorship...17 Behavior and Feeding Habits Olives + Insects + Water...18 Weight Comparison Egg masses, Egg, Relative Humidity/Temperature

5 Olive Observations...19 Survivorship.20 Behavior and Feeding Habits. 20 -Insects + Water Weight Comparison.22 Egg masses, Eggs, Relative Humidity/Temperature Olive Observations Survivorship Insects + Olive Leaves...23 Weight Comparison Egg Masses, Eggs, Relative Humidity/Temperature Olive Observations. 23 Survivorship Insects + Olive Leaves + Olives. 24 Weight Comparison Egg masses, Egg, Relative Humidity/Temperature Olive Observations. 24 Survivorship...25 Evaporation Rate and Feeding Damage.25 Survivorship Reproduction..26 Discussion Survivorship..28 -Olive Observations and damage 29 -Relative Humidity and Temperature.30 -Reproduction.31 -Behavior and Feeding Habits 31 - For Olive growers...32 Table 1. Olive treatment descriptions 33 Figure 1. CDFA distribution map of brown marmorated stink bug Figure 2. (A, B, C, D). Images of Olive damage... 35

6 Figure 3. Brown marmorated stink bug survivorship (Insects + Olives) Figure 4. Brown marmorated stink bug survivorship (Olives + Insects + Water) Figure 5. Brown marmorated stink bug survivorship (Insects + Water). 38 Figure 6. Brown marmorated stink bug survivorship (Insects + Olive Leaves)..39 Figure 7. Start, end, and difference in olive weight for trial #1..40 Figure 8. Start, end, and difference in olive weight for trial #2..41 Figure 9. Start, end, and difference in olive weight for trial # Figure 10. Start, end, and difference in olive weight for trial # Figure 11. Number of egg masses and total amount of eggs (Olive + Insects).. 44 Figure 12. Number of egg masses and total amount of eggs (Olive+ Insects+Water...45 Figure 13. Number of egg masses and total amount of eggs (Insects + Water)...46 Figure 14. Number of egg masses and total amount of eggs (Insects+ Olive Leaves)..47 Figure 15. Insect contacts with olives in trial # Figure 16. Insect contacts with olives in trial # Figure 17. Insect contacts with olives in trial # Figure 18. Insect contacts with olives in trial # Figure 19. Olive Damage from BMSB 52 Work Cited/References.. 53

7 Introduction: The brown marmorated stink bug, Halyomorpha halys (Pentatomidae), is an invasive insect in the United States that originated in East Asian countries such as Japan, China, Korea, and Taiwan (Welty et al., 2008). Brown marmorated stink bugs were first introduced to the Eastern United States in the late 1990 s. This species of stink bugs were first reported in Allentown, Pennsylvania in 1996 (Zhu et al., 2012). In addition to being excellent flyers, stink bugs are sometimes known as hitch hikers, meaning they can be transported by human activity, which allows them to spread quickly compared to other insects. As of 2015, they have been reported in 42 of the 50 states and are expected to spread to the remaining states (Figure 1). The insect was found during mid-2000 in Los Angeles County and, in 2013, was detected in Sacramento. According to the California Department of Food and Agriculture (2015) these insects have reproducing populations in Butte, Los Angeles, Sacramento, San Joaquin, Santa Clara, and Sutter Counties (Figure 1). They have been intercepted in over 20 counties in California and are expected to continue to spread. Besides being an urban nuisance, stink bugs are an agricultural pest that have caused millions of dollars of agricultural damage on the East Coast by feeding on crops such as stone fruits, field tomatoes, peppers, hazel nuts, and apples (Welty et al., 2008). Once this insect branches out from urban to agricultural areas in California, the results could be worse than the East Coast. Unlike California, the East Coast has a much more severe climate that limits what crops can be grown. Some of the favorite hosts of these insects are fruits, vegetables, and ornamental trees that are common in California and grow where stink bugs have currently established themselves. Several distinct features stand out about the brown marmorated stink bug compared to other species of stink bugs. BMSB can be distinguished by the white bands on their charcoal colored antenna, a smooth anterior pronotal margin in the shoulder region, and the banding pattern on the abdominal margin (Medal et al., 2013). The underside of the abdomen can be 1

8 brightly colored or have speckled pigment (Medal et al., 2013). Generally, the female adults are larger than male adults, ranging from millimeters in length with the males ranging from 7-10 millimeters in length (Medal et al., 2013). Depending on the stage of adulthood, BMSB can be white, grey, or tan. Each of the five instars of their life cycle, from nymphs to adults has the potential to cause damage. This is part of the reason why these insects are such a threat (Medal et al., 2013). Each stage of instar, expect the last or 5 th instar, last about one week before evolving to the next. The 5 th instar, or final stage to adulthood, lasts about two weeks (Medal et al., 2013). Once adults, females on live on average 9 to 16 weeks and the males survive on approximately 8 to 18 weeks in a controlled environment. Typically, the male to female ratio of emerged brown marmorated stink bugs is 0.9:1 (Medal et al., 2013). Females have the potential to lay over 400 eggs in their lifetime and usually lay around 28 eggs per egg mass. BMSB damage fruit, vegetables, pods, buds, and stems of plants by feeding and sucking fluid out of them. They pierce the skin of what they are feeding on with their proboscis and inject saliva enzymes that break down the inside of the fruit or vegetable and the stink bugs are able to suck out the remaining fluid (Kuhar and Kamminga 2012). The brown marmorated stink bug secretes two different types saliva that consist of a watery and sheath saliva (Peiffer and Felton 2014). The sheath saliva is used when feeding and causes the areas that have been pierced by the proboscis to remain open on the fruits and vegetables for continued feeding. This often results in bleeding or leaking of fluid and, as a result, fruits and vegetables are more susceptible for pathogenic bacteria to enter them (Peiffer and Felton 2014) (Figure 2A). The damage caused by these insects usually appears as corking, discoloration, or collapsing (Kuhar and Kamminga 2012) (Figure 2 B, C, and D). Crops are attacked the most during July through September, as insects aggregate in early fall and then reside in homes and buildings during the winter months 2

9 (Kuhar and Kamminga 2012). It is not until the spring that brown marmorated stink bugs return to the outdoors and start reproducing and damaging crops. The brown marmorated stink bug poses more of a threat than other species. They have limited natural enemies to control the population as they are not native to the United States. Trissolcus mitsukurii and Trissolcus japonicas are parasitoids native to Asia that are currently being studied to be released to help control these insects; this has not been done yet on the West Coast (Durham and O Brien 2013). With fewer natural enemies, it gives stink bugs the capability of having a greater number of individuals produced each generation. They have been known to have one to five generations a year depending on the temperature, photoperiod, and climate. With a larger number of generations per year, it gives brown marmorated stink bugs the opportunity to feed on more crops for longer periods of time (Zhu et al., 2012). Lastly, they are resistant to commonly used pesticides, which makes it difficult to control the population of these insects (Bergmann and Raupp, 2014). Spraying intervals will have to be more frequently applied to crops to combat these stink bugs while still seeing high crop damage, which could lead to more problems for California agriculture (Bergmann and Raupp, 2014). California Agriculture: California is one of the top agriculture producing states in the United States and is made up of over 80,000 farms (Medina 2014). It is the nation s largest exporter of agricultural commodities due to the unique environment types, dependable water supply, soil, and moderate Mediterranean climate. These factors allow a wide variety of crops to be grown year round. In 2013, over 13.7 billion dollars worth of agricultural exports were shipped around the world (Medina, 2014). Many of these exports are made up of California s massive specialty crops and bring in billions of dollars annually. Crops include almonds, avocados, cherries, grapes, olives, 3

10 pistachios, and many different types of citrus (Tolomeo et al., 2015). The top agriculture producing counties in California are Fresno, Kern, Merced, Monterey, and Tulare (Medina, 2014). Since the brown marmorated stink bug is relatively new to California, it has not established reproducing populations in these major agricultural areas; rather is considered a nuisance pest because of their growing population and ability to reside over winter in homes (O Brien, 2009). The unique agricultural industries of California could be at risk from stink bugs as well as the already known crops on the East coast that are vulnerable to brown marmorated stink bug damage. California has the potential to exceed the economic damage that was done in the Mid- Atlantic States (Nik Wiman et al. 2015). Many scientist from University of California Agriculture and Natural Resources (UC Davis), UC Riverside Center for Invasive Species Research, University of California Berkeley, the California Department of Food and Agriculture (CDFA), and United States Department of Agriculture (USDA) are working together to see how these insects will affect western agriculture specialty crops. This report focuses on the brown marmorated stink bug and determines whether they will have the potential to impact the expanding California olive industry. California Olive Industry: Olives were first introduced to California in the late 1700 s and were farmed commercially by the late 1800 s throughout the central valley (Vossen, 2007). Most olives are used as table olives while a small percentage are used for olive oil, although over the last ten years California olive oil has become more popular. The demand for California olive oil has doubled in the last 15 years with over 400,000 gallons of olive oil produced yearly (Vossen, 2007). Currently, olives are one of the smaller specialty crops grown in California, but are 4

11 gaining popularity and have the potential to grow as the wine industry did (Vossen, 2007). According to the 2002 Census of Agriculture, California had 40,000 acres of olives grown on over 1500 farms and leads the nation in olive production by producing 70-80% of the ripe olives consumed in the United States (Tolomeo et al., 2015). There are five main varieties of olives grown in California: Mansanillo, Sevillano, Mission, Ascolano, and Barouni, all of which may be affected by brown marmorated stink bugs (Vossen, 2007). The leading olive producing counties are Butte, Glenn, Tehama, and Tulare (Tolomeo et al., 2015). Since none of these counties have been exposed to brown marmorated stink bugs, the consequences of BMSB feeding on olives are currently unknown (Figure 1). The olive industry in California already faces problems caused by the invasive olive fruit fly and could possibly be threatened with a new insect (Figure 2A). The brown marmorated stink bug has the potential to cause devastating damage to olives that could result in economic damage to olive growers once they spread. Objectives The goal of this project is to determine if BMSB will be able to survive, reproduce, and damage olives. It will also determine at what time of the day these insects feed and it will be determined if the insects will receive the correct amount of nourishment from olives to be able to survive and reproduce. Brown marmorated stink bug feeding habits will be captured using digital imaging. If the insects are able to survive, the amount of damage they cause to the olives will be evaluated to determine if olives would be acceptable to processors once harvested. The hypothesis to be tested is: BMSB will be able to survive and reproduce on olives and cause enough damage to have a negative economic impacts on olive growers. 5

12 The insects are new to the West Coast and California is the only state that grows table olives, making this one of the first experiments done regarding stink bugs and olives. This study will benefit olive growers by expanding knowledge and awareness of how this new invasive pest can potentially be a threat to the industry. It will also be beneficial to professionals and the public because it will educate them on how this insect has the potential to damage different varieties of agriculture throughout California. This information will alert the olive industry to the potential impact this new pest can bring to their industry. Methods: The experiment took place in the California Department of Food and Agriculture, biological Control Quarantine building and lasted from June 29 th, 2015 to October 7 th, Four, two week trials, were set up, observed, and documented. The trials consisted of four to seven treatments designed to measure the ability of brown marmorated stink bugs to survive and produce eggs using combinations of olive fruit and water. Various combinations of stinkbugs, olives, and plant material were used to address the question of what is necessary for brown marmorated stink bugs to survive on olives (Table 1). Each treatment was replicated four times for every trial. The test arena for each replicate was a plastic petri dish with a 3 cm diameter screen covered hole cut into its top half to allow airflow. Additional time was added between trials to allow olives to mature on trees. Female brown marmorated stink bugs were used throughout the length of the experiment. There were two sources of brown marmorated stink bugs: either coming from the lab culture (trials 3 and 4), from a local community garden (trial 2), or a mix of both (trial 1). Each replicate contained four olives in every treatment and trial. The olives were collected from a randomly chosen tree near a bike trail in Davis, California. The insects and olives that were used were all collected within 24 to 48 hours of starting each trial. 6

13 Two to five insects were used depending on the specific treatment. Cotton swabs were added to the treatments with ml of water in each. Water was added to the cotton swabs as needed throughout the duration of each trial to remain wet. Every trial was observed Monday, Wednesday, and Friday for a two-week period. Observations were taken on olive damage (bleeding and piercing), color (purple and green), texture (smooth or shriveled), number of egg masses, and number of dead and live insects. The start and end weight was recorded for every trial. During the last two trials the weight of the olives was recorded half way through the trial for more accuracy. The egg masses were counted then removed each time the experiment was checked. Temperature and relative humidity were recorded using Watch Dog Model 250 and Hobo Model (UX100-03) data loggers for trial three and four. A Wingscape camera was set up to take pictures every five to ten minutes to capture the insect s behavior throughout different times of the day for the majority of each trial. In the entire experiment there was a total of seven different treatments; Olives, olive leaves, olive stems, water soaked cotton, and stink bugs were added to each petri dish using various combinations and sealed with parafilm (Table 1.). 1. Control #1: Olives Used to determine if olive tissue broke down in the absence of any impact from brown marmorated stink bugs. 2. Control # 2: Olives + Water Water was added to olives to control for moisture added to other treatments listed below. 3. Insects + Water Used to determine brown marmorated stink bug survivorship on water alone. 4. Insects + Olives Used to determine how long brown marmorated stink bugs could survive on just olives as a food source. 7

14 5. Insects + Olives + Water Used to determine if survivorship increased in the presence of both, water and olives as a food source. 6. Olive Leaves + Insects Used to determine if brown marmorated stink bugs could use olive stems or leaves as a water source, in addition to olives. 7. Olive Leaves + Insects + Olives Determine if survivorship increased using leaves plus water plus olives as a food source. Trial # 1 Trial # 1 was set up June 26, 2015 and run two weeks until July 10, Observations were recorded on June 29 th, July 1 st, 3 rd, 6 th, 8 th, 10 th. Three treatments were used in this trial: Control #1 Olives, Olives + Insects, and Olives + Insects + Water. The camera was set up on July 6 th and was left on until the end of trial taking pictures every five minutes. The start and end weights of the olives were recorded on the first day, June 26 th, and the last day, July 10 th. Trial #2 Trail # 2 was set up on July 17, 2015 and run for two weeks, ending July 31, Observations were recorded on July 20 th, 22 nd, 24 th, 27 th, 29 th, and 31 st. Due to the lack of brown marmorated stink bugs at the time this trial was set up, only four insects were placed in each replicate. There was a total of five different treatments used in this trial: Control #1 Olives, Control # 2 Olives + Water, Olives + Insects, Insects + Water, and Olives + Insects + Water. The Camera was set up on July 17 th and was left on until July 28 th taking pictures every five minutes. The start and end weights of the olives were recorded on the first day, July 17 th, and the last day, July 31 th. 8

15 Trial #3 Trial #3 was set up on September 7, 2015 and run for two weeks, ending September 21, Observations were recorded on September 9 th, 11 th, 14 th, 16 th, 18 th, and 21 st. There were six treatments used in this trial: Control #1 Olives, Control # 2 Olives + Water, Olives + Insects, Insects + Water, Olives + Insects + Water, and Olive Leaves + Insects. The Camera was set up on September 7 th and was left on until September 21 st, taking pictures every five minutes. The starting weight of the olives was recorded on September 7 th, the half way weight was recorded on September 16 th, and the end weight was recorded on September 21 st. Relative humidity and temperature were recorded in the different treatments throughout the trial with Watch Dog Model 250 and Hobo Model UX data loggers. Trial #4 Trial #4 was set up on September 23, 2015 and run for two weeks, ending October 7, Observations were recorded on September 25 th, 28 th, 30 th, October 2 nd, 5 th, and 7 th. There were seven treatments used in this trial: Control #1 Olives, Control # 2 Olives + Water, Olives + Insects, Insects + Water, Olives + Insects + Water, Olive Leaves + Insects, and Olive Leaves + Olives + Insects. The camera was set up on September 23 rd and was left on until October 7 th, taking pictures every five minutes. The starting weight of the olives was recorded on September 23 rd and the end weight was recorded on October 7 th. Relative humidity and temperature were recorded in the different treatments throughout the trial using Watch Dog Model 250 and Hobo Model UX data loggers. 9

16 Weight Comparison and Damage The two controls, Olives, and Olives + Water were used to compare with Olives + Insects and Olives + Insects + Water to determine weight differences and observable damage at the end of the trials to determine whether brown marmorated stink bugs caused damage to the olives. The olive weights at the end of each trial were measured to determine if the insects were feeding on the olives. It was assumed that if olives were losing weight, they were losing it due to evaporation and the insects feeding on them. The average evaporation rate was determined by subtracting the average weight differences from start to end of all four replicates of Olives from Olives + Water. The amount of weight lost due to the insects feeding on olives was determined by subtracting the average difference in olive weights from start to end for all four replicates at the end of the trial of Olives + Insects + Water from Olives + Water. At the end of the trial, olives were taken from each of the treatments and dissected and photographed under a microscope using Wingscape software (Figure 2 C and D). All the olives had been previously exposed to the invasive olive fruit fly, therefore other olives that were collected at the same time and not exposed to the stink bugs were dissected to compare the damage the olive fruit fly can have on olives to the damage caused by brown marmorated stink bugs (Figure 2A). Behavior and Feeding Habits Digital imaging was used throughout the duration of each trial to record behaviors and feeding habits of the brown marmorated stink bugs. Pictures were taken every ten minutes for most of each trial. It was assumed that if the insects were on the olives, they were probing and feeding on them. Feeding on olives was measured by counting the number of frames showing brown marmorated stink bugs making contact with olives. Four time blocks of one hour intervals per day were randomly chosen to determine the amount of insects that came in contact with 10

17 olives in each of the replicates. For every trial this information was recorded and averaged for these time frames. The number of frames was divided by the number of images used (in this case 6 images per hour) to adjust the differences of times the petri dish was imaged each hour. For example, if five images are shot each hour versus ten, the bugs are twice as likely to be seen feeding. Also, the number of frames was divided by the number of insects seen making contact with olives in each frame. This was recorded in the beginning of the trial to simplify the amount of survivorship taking place in each replicate. Example: if in one hour, six images were taken and all four replicates were viewable the sample size would be 24 (6 pictures per hour x 4 replicates = 24 replicates). If 15 of the replicates all had one bug feeding on the olives and the other 9 replicates had zero bugs feeding on the olives the equation would look like this: = 15/24(adjusted sample size) = bug contacts per hour. Reproduction, Relative Humidity, and Temperature The experiment was checked every day and the number of egg masses laid in each treatment and replicate were recorded. The specific number of eggs laid per egg mass was counted under a microscope for accuracy. The egg masses were removed and placed in the freezer to kill and dispose of at a later date. The Watch Dog Model 250 and Hobo Model UX data loggers were moved randomly to get a recording of the humidity and temperature for each of the treatments in trial #3 and #4. It was started in trial #3 due to a larger number of insects dying in the beginning of the trial in the treatments containing water. The 11

18 devices documented the data every one, five, and fifteen minutes depending on the device. At the end of the trial, the start and end times were recorded and each entry was averaged over the period of time it was in each individual treatment. Results: Control #1- Olives: Weight Comparison of Trials: The average start weight for olives in the treatment Control #1 Olives in trial #1 was 4.98 grams, the average end weight was 3 grams, and the average difference was 1.9 grams. In trial #2 the average start weight was 4.91 grams, the average end weight was 2.87 grams, and the average difference was 2.04 grams. In trial #3 the average start weight was 6.52 grams, the average end weight was 5.04 grams, and the average difference was 1.48 grams. In trial #4 the average start weight was 5.4, the average end weight was 4.12 grams, and the average difference was 1.22 grams. (Figures 7, 8, 9, and 10). Relative Humidity and Temperature: Relative humidity and temperature were not recorded for trials 1 and 2. For trial 3, relative humidity was not recorded in the Control # 1 Olives treatment. For trial 4, relative humidity and temperature were recorded from 9/25/15 at 9:06 A.M. until 10/2/15 at 8:35 AM. The average temperature over that period of time was degrees Celsius and the average RH was

19 Olive Observations: Trial #1: The olives started out being green with a smooth texture. Throughout the trial they were green with developing black spots and had more of a shriveled texture. As the trial continued, the olives got more black spots and eventually by the end of the trial they were completely black and were very shriveled. Trial #2: The olives started out being green with a smooth texture. Black spots did not develop on the olives until the last day of the trial. Olives continually shriveled throughout the course of the trial. Trial #3: The olives started out mostly being purple, and a few were still green and all had a smooth texture. As the trial progressed, the olives went from being green/purple to completely purple. The bigger olives also were not as shriveled as the smaller olives at the end of the trial. Trial #4: The olives starting out being green/purple and a couple of completely green olives. By the end of the trial, all the olives were completely purple, had a shriveled texture, and the bigger olives did not have as much shriveling on them as the smaller olives. Survivorship: No insects were used in this treatment. 13

20 Control #2- Olives + Water: Weight Comparison of Trials: The average start weight for olives in the treatment Control #2 Olives + Water in trial #1 was not recorded. In trial #2 the average start weight was 4.51 grams, the average end weight was 3.91 grams, and the average difference was 0.61 grams. In trial #3 the average start weight was 6.63 grams, the average end weight was 5.96 grams, and the average difference was 0.67 grams. In trial #4 the average start weight was 5.42, the average end weight was 5.21 grams, and the average difference was 0.21 grams. (Figures 7, 8, 9, 10) Relative Humidity and Temperature: Relative humidity and temperature were not recorded for trials 1 and 2. For trial #3 the relative humidity and temperature were recorded from 9/18/2015 at 9:00 A.M. to 9/21/2015 at 8:00 A.M. The average relative humidity was and the average temperature was degrees Celsius. For trial 4, relative humidity and temperature were recorded from 10/2/15 at 8:36 A.M. until 10/7/15 at 1:00P.M. The average temperature over that period of time was degrees Celsius and the average RH was Olive Observations: Trial #1: Not Recorded Trial #2: Olives were completely green and had a smooth texture throughout the entire trial. Towards the end of the trial, the olives were just starting to shrivel. Trial #3: Olives were mostly green with very little purple with a smooth texture for the beginning of the trial. Throughout the trial they slowly turned purple and by the end of the trial 14

21 were completely purple with very little shriveling. The bigger olives had no shriveling on them and the smaller ones did. Trial #4: Olives were purple and green and had a smooth texture in the beginning of the trial. By the end of the trial they were all completely purple and still remained smooth. Some of the olives had mold on them. Survivorship: No insects were used in this treatment. Olives + Insects: Weight Comparison of Trials: The average start weight for olives in the treatment Olives + Insects in trial #1 was 4.9 grams, the average end weight was 4.5 grams, and the average difference was 0.36 grams. In trial #2, the average start weight was 5.08 grams, the average end weight was 2.98 grams, and the average difference was 2.11 grams. In trial #3, the average start weight was 6.22 grams, the average end weight was 4.37 grams, and the average difference was 1.85 grams. In trial #4, the average start weight was 5.65, the average end weight was 4.0 grams, and the average difference was 1.65 grams. (Figures 7, 8, 9, and 10) Egg Masses, Eggs, Relative Humidity, and Temperature: In trial #1, there were two egg masses found with a total of six eggs. In trial #2, there were zero egg masses found and a total of zero eggs. In trial #3, there were five egg masses 15

22 found with a total of 123 eggs. In trial #4, there were zero egg masses found and a total of zero eggs. (Figure 11). The temperature and relative humidity were not recorded for trials #1 and #2. For trial #3, the relative humidity and temperature were recorded from 9/14/2015 at 8:39 A.M. to 9/18/2015 at 8:53 A.M. The average relative humidity was and the average temperature was degrees Celsius. For trial #4, the relative humidity and temperature were recorded from 9/29/2015 at 8:00 A.M. to 10/7/2015 at 1:00 PM. The average relative humidity was and the average temperature was degrees Celsius. Olive Observations: Trial #1: Olives were completely green in the start of the trial. Throughout the trial the olives became shriveled and started to have black spots on them. Some of the olives clearly had more black spots on them than others and the spots increased as the trial continued. At the end of the trial, olives had big black spots on them, some were completely black, and they were all very shriveled. Trial #2: Olives were all green and smooth in the start of the trial. They developed tiny black spots and continued to shrivel as the trial progressed. Trial #3: Olives were mostly green with a little purple on them and had a smooth texture at the start of the trial. By the end of the trial olives were purple, and shriveled, but not as shriveled as the previous trials. Trial #4: There was a mix of green and purple olives and all were smooth in the beginning of the trial. By the end, all olives were purple and just starting to shrivel. Two maggots were found in the one of the replicates that burrowed out of the olives. 16

23 Survivorship: In trial #1 of Olives + Bugs, the mean of surviving insects was 0.46 with an average standard deviation of 0.38 and a standard error of In trial #2, the mean of surviving insects was 1.25 with an average standard deviation of 0.87 and a standard error of In trial #3, the mean of surviving insects was 2.42 with a standard deviation of 0.4 and a standard error of 0.2. In trial #4, the mean of surviving insects was 2.86 insects with a standard deviation of 0.53 and a standard error of (Figure 3) Behavior and Feeding Habits: Bug Contact average per hour (Standard Deviation) Trial #1: Not recorded, all insects were dead by the time the camera was set up. Trial #2 (Figure 16): 6:00 AM: 0.79 bug contacts/hour (0.51) 10:00 AM: 0.66 bug contacts/hour (0.86) 2:00 PM: 0.25 bug contacts/hour (0.44) 6:00 PM: 0.29 bug contacts/hour (0.46) Trial #3 (Figure 17): 6:00 AM: 1.5 bug contacts/hour (0.79) 10:00 AM: 1.25 bug contacts/hour (1.21) 2:00 PM: 3.08 bug contacts/hour (2.02) 17

24 6:00 PM: 1.75 bug contacts/hour (0.75) Trial #4 (Figure 18): 6:00 AM: 0.08 bug contacts/hour (0.28) 10:00 AM: 0.33 bug contacts/hour (0.56) 2:00 PM: 1.45 bug contacts/hour (1.17) 6:00 PM: 1.29 bug contacts/hour (0.91) The average was taken for each of these times, the highest period of feeding activity was at 2:00 PM, with 1.59 bug contacts per hour. Next was at 6:00 PM with 1.11 bug contacts per hour. The two afternoon times had more feeding activity on them than the morning hours. At 7:00 AM the average bug contact throughout the trials was 0.77 bug contacts per hour. The least amount of bug activity was at 10:00 AM with 0.74 bug contacts per hour. Olives + Insects + Water: Weight Comparison of Trials: The average start weight for olives in the treatment Olives + Insects + Water in trial #1 was 5.5 grams, the average end weight was 4.5 grams, and the average difference was 0.91 grams. In trial #2, the average start weight was 4.51 grams, the average end weight was 3.68 grams, and the average difference was 0.82 grams. In trial #3, the average start weight was 5.95 grams, the average end weight was 5.39 grams, and the average difference was 0.56 grams. In trial #4 the average start weight was 6.04, the average end weight was 5.33 grams, and the average difference was 0.71 grams. (Figure 7, 8, 9, and 10) 18

25 Egg Masses, Eggs, Relative Humidity, and Temperature: In trial #1, there were 14 egg masses found with a total of 308 eggs. In trial #2, there were 11 egg masses found and a total of 201 eggs. In trial #3, there were 12 egg masses found with a total of 283 eggs. In trial #4, there were two egg masses found and a total of 48 eggs. (Figure 12). The temperature and relative humidity were not recorded for trials #1 and #2. For trial #3 the relative humidity and temperature were recorded from 9/9/2015 at 9:00 A.M. to 9/14/2015 at 8:00 A.M. The average relative humidity was and the average temperature was degrees Celsius. For trial #4 the relative humidity and temperature were recorded from 9/29/2015 at 8:00 A.M. to 10/7/2015 at 1:00 PM. The average relative humidity was and the average temperature was degrees Celsius. Olive Observations: Trial #1: Olives started out being green and smooth for the beginning of the trial. By the forth observation day, they had black spots on them and were starting to shrivel. At the end of the trial the olives were still green, had black spots on them, and were all just starting to shrivel. Trial #2: Olives were green, had a smooth texture, and all contained small black spots on them for the entire trial. Trial #3: Olives started out being green with a little purple, smooth texture, and no observable damage on them. By the end of the trial the olives were all purple but still had a smooth texture 19

26 to them. Some of the olives had some discoloration on them and developed mold, which was not observed in previous trials. Trial #4: Olives where mostly green with a small amount of purple on them in the start of the trial. By the end, olives were completely purple and had a smooth texture to them. Survivorship: In trial #1 of Olives + Insects + Water, the mean of surviving insects was 3.83 with an average standard deviation of 0.33 and a standard error of In trial #2, the mean of surviving insects was 2.67 with an average standard deviation of 0.77 and a standard error of In trial #3, the mean of surviving insects was 2.96 with a standard deviation of 0.75 and a standard error of In trial #4, the mean of surviving insects was 4.21 insects with a standard deviation of 0.80 and a standard error of (Figure 2). Behavior and Feeding Habits: Bug Contact average per hour (Standard Deviation) Trial #1 (Figure 15): 6:00 AM: 0.75 Bug contacts/hour (0.61) 10:00 AM: 0.29 Bug contacts/hour (0.47) 2:00 PM: 0.41 Bug contacts/hour (0.65) 6:00 PM: 0.91 Bug contacts/hour (0.65) Trial #2 (Figure 16): 6:00 AM: 0.5 Bug contacts/hour (0.51) 20

27 10:00 AM: 0.63 Bug contacts/hour (0.49) 2:00 PM: 0.91 Bug contacts/hour (0.58) 6:00 PM: 0.79 Bug contacts/hour (0.72) Trial #3 (Figure 17): 6:00 AM: 1.1 Bug contacts/hour (0.87) 10:00 AM: 1.1 Bug contacts/hour (0.63) 2:00 PM: 1.7 Bug contacts/hour (0.57) 6:00 PM: 0.7 Bug contacts/hour (0.54) Trial #4 (Figure 18): 6:00 AM: 1.08 Bug contacts/hour (0.92) 10:00 AM: 0.08 Bug contacts/hour (0.28) 2:00 PM: 1.41 Bug contacts/hour (1.1) 6:00 PM: 2.1 Bug contacts/hour (0.94) The average was taken for each of these times, the highest period of feeding activity was at 6:00 PM, with 1.13 bug contacts per hour. Next was at 2:00 PM with 1.11 bug contacts per hour. The two afternoon times had more feeding activity on them than the morning hours. At 7:00 AM the average bug contact throughout the trials was 1.03 bug contacts per hour. The least amount of bug activity was at 10:00 AM with 0.53 bug contacts per hour. 21

28 Insects + Water: Weight Comparison of Trials: The average start weight for olives in the treatment Insects + Water in trial #1, #2, #3, and #4 were not recorded since there was no olives used in this treatment. Egg Masses, Eggs, Relative Humidity, and Temperature: In trial #1, this treatment was not conducted and there is no results for egg masses and number of eggs. In trial #2, there were 9 egg masses found and a total of 41 eggs. In trial #3, there were 7 egg masses found with a total of 180 eggs. In trial #4, there were 2 egg masses found and a total of 53 eggs. The temperature and relative humidity were not recorded for trials #1 and #2 (Figure 13). For trial #3, the relative humidity and temperature were not recorded for trial #3. For trial #4, the relative humidity and temperature were recorded every one minute from 9/29/2015 at 8:00 A.M. to 10/7/2015 at 1:00 PM. The average relative humidity was and the average temperature was degrees Celsius. Olive Observations: No olives were used for this treatment. Survivorship: In trial #1 of Insects + Water was not conducted and there are no results. In trial #2, the mean of surviving insects was 1.5 with an average standard deviation of 0.75 and a standard error of In trial #3, the mean of surviving insects was 2.0 with a standard deviation of

29 and a standard error of In trial #4, the mean of surviving insects was 3.92 insects with a standard deviation of 0.57 and a standard error of (Figure 5). Insects + Olive Leaves: Weight Comparison of Trials: There were no olives used in this treatment. Egg Masses, Eggs, Relative Humidity, and Temperature: In trial #1 and #2 this treatment was not conducted and there is no results for egg masses and number of eggs. In trial #3 there were four egg masses found and a total of 114 eggs. In trial #4 there were 4 egg masses found with a total of 81 eggs. In trial #4 there were 2 egg masses found and a total of 53 eggs. (Figure 14). The temperature and relative humidity were not recorded for trials #1 and #2. For trial #3, the relative humidity and temperature were recorded from 9/15/2015 at 8:39 A.M. to 9/21/2015 at 8:00 A.M. The average relative humidity was and the average temperature was degrees Celsius. For trial #4, the relative humidity and temperature were not recorded. Olive Observations: No olives were used in this treatment. Survivorship: In trial #1 of Insects + Water was not conducted and there are no results. In trial #2, the mean of surviving insects was 1.5 with an average standard deviation of 0.75 and a standard 23

30 error of In trial #3, the mean of surviving insects was 2.0 with a standard deviation of 0.73 and a standard error of In trial #4, the mean of surviving insects was 3.92 insects with a standard deviation of 0.57 and a standard error of (Figure 6). Insects + Olive Leaves + Olives: Weight Comparison of Trials: The olives used in this trial were attached to the stem and were not weighed. Egg Masses, Eggs, Relative Humidity, and Temperature: In trial #1, #2, and #3 this treatment was not conducted and there is no results for egg masses and number of eggs. In trial #4, there were 1 egg masses found and a total of 28 eggs. Relative humidity and temperature were not recorded for trial number 3. For trial #4, the relative humidity and temperature were recorded from 9/25/2015 at 8:00 A.M. to 10/7/2015 at 1:00 PM. The average relative humidity was and the average temperature was degrees Celsius. Olive Observations: Not recorded for trials #1, #2, and #3 Trial #4: Olives started out as green and starting to turn purple with a smooth texture. By the end of the trial the olives were all purple and starting to shrivel. 24

31 Survivorship: In trial #1 of Insects + Water was not conducted and there are no results. In trial #2, the mean of surviving insects was 1.5 with an average standard deviation of 0.75 and a standard error of In trial #3, the mean of surviving insects was 2.0 with a standard deviation of 0.73 and a standard error of In trial #4, the mean of surviving insects was 3.92 insects with a standard deviation of 0.57 and a standard error of Behavior and Feeding Habits: This information was not recorded for this treatment. Evaporation Rate and Feeding Damage from Brown marmorated stink bugs: The average evaporation rates for each two week trial are listed: Trial #1: Not recorded Trial #2: 1.43 grams Trial #3: 0.81 grams Trial #4: 1.05 grams As each trial was run, the stink bugs fed more on the olives causing an overall trend of increasing damage. In trial #1, the Olives + Insects treatment lost 0.1 grams more when compared to the Control #1: Olives treatment. The Olives + Insect + Water treatment was unable to be compared to any control since the Control #2 treatment was not used in trial #1 (Figure 7). In trial #2, the Olives + Insects treatment lost 0.05 grams more when compared to the Control #1: Olives treatment. The Olives + Insect + Water treatment lost 0.21 grams more when compared to the Control #2: Olives + Water treatment (Figure 8). In trial #3, the Olives + Insects 25

32 treatment lost 0.37 grams more when compared to the Control #1: Olives treatment. The Olives + Insect + Water treatment lost 0.11 grams more when compared to the Control #2: Olives + Water treatment (Figure 9). In trial #4, the Olives + Insects treatment lost 0.43 grams more when compared to the Control #1: Olives treatment. The Olives + Insect + Water treatment lost 0.5 grams more when compared to the Control #2: Olives + Water treatment (Figure 8). Survivorship: The survivorship trend throughout the four, two week trials varied for each of the trials. In the treatment, Bugs + Olives, survivorship increased with each continuing trial. In trial #1, the survivorship rate at the end of the trial was 0.45 insects and at the end of trial # 4, the survivorship rate was 2.85 (Figure 3). In the treatment, Insects + Olives + Water, survivorship remained around the same at the end of all trials but increased slightly. At the end of trial #1, survivorship was 3.8 insects and at the end of trial #4, survivorship was 4.2 insects (Figure 4). Survivorship for the treatment, Insects + Water, survivorship increased over the course of the four trials. In trial #2 (not conducted in trial #1), survivorship was 1.5 insects and at the end of trial #4, survivorship was 3.91 insects (Figure 5). The survivorship in the treatment, Insects + Olive Leaves, survivorship decreased. In trial #3 (not conduced in trial #1 and #2), survivorship was 1.2 insects and at the end of trial #4, the survivorship rate was 0.88 insects (Figure 6). Reproduction in each treatment: In the treatment, Olives + Insects, egg masses were only produced in trial #3. The Olives + Insects + Water treatment produced egg masses for each trial, but in trial #4, egg mass production decreased significantly compared to the other trials. Egg mass production also declined from trial to trial in the Insects + Water treatment. The egg mass production in the 26

33 Olive Leaves + Insects treatment gradually increased from trial # 3 to trial #4. (Figures11, 12, 13, and 14). Discussion: The results from the four, two week trials indicated that brown marmorated stink bugs are able to survive on olives. Since these insects were able to survive, they were also able to reproduce and cause internal and external damage to the olive fruit. Also, with digital imaging, it was determined that brown marmrorated stink bugs were more active and fed more on the olives in the afternoon than the morning hours. Based on these findings it was observed that: 1.) Brown marmorated stink bugs are able to survive in some treatments better than others because the water and more mature olives provided enough nourishment; 2.) Brown marmorated stink bugs are able to survive better on more mature olives since they have more nutrients present; 3.) An alternative water source to olives alone increases the survival rate of brown marmorated stink bugs regardless of maturity of olives; 4.) Brown marmorated stink bugs caused damage to the inside and outside of olives; 5.) Brown marmorated stink bugs caused weight loss to olives; 6.) Relative humidity and temperature did not change the survivorship on brown marmorated stink bugs; 7.) Brown marmorated stink bugs were able to reproduce better in some treatments than others because of the amount of nourishment and daylight; 27

34 8.) Insects were most active and fed more in the afternoon compared to the morning hours. Survivorship in treatments: Four treatments were used that involved the insects: 1.) Olives + Insects 2.) Olives + Insects + Water 3.) Insects + Water 4.) Insects + Olive Leaves. In the treatment Olives + Insects, the survivorship rate in trial #1 was low, and as each trial was conducted the survivorship rate increased. The insects were not able to survive on the younger olives in the early trials. Once the olives had time to mature, the insects were able to receive the correct amount of nourishment to survive. In the treatment Olive + Insects + Water, the insects had a high survival rate throughout all four trials. The only difference was in the later trails, with more mature olives, the insects had a more consistent survival rate day to day than in the earlier trials. This could have been a result from the insects gaining better nourishment from more mature olives than in the earlier trials. The addition of a water source helped the insects survive better than without. In the treatment, Insects + Water, the insect s survival rate steadily increased from trial to trial. This could have been a result of the later generations of insects, that are born closer to the over wintering months, do not need as much nourishment to survive. The last treatment, Olive Leaves + Insects was only done in two trials, making it difficult to see a trend. In the two trials it was done, survivorship decreased and the insects were unable to get the correct amount of nourishment from the leaves and stems of the olives. Based on the results, it is evident that brown marmorated stink bugs are 28

35 able to survive on mature olives and can also survive on younger olives with an alternate water source. Feeding Damage to Olives: It was anticipated that the brown marmorated stink bugs would be able to cause damage to olives by feeding on them. In trials #1, 2, 3, and 4, the treatment, Olives + Insects always lost more weight than the treatment, Control #1: Olives (Figures 7, 8, 9, and 10). In trials # 2 and 4, the treatment, Olives + Insects + Water always lost more weight than the treatment, Control: Olives + Water (Figure 8 and 9). In trial #3, it was the opposite but was not much of a difference (Figure 9). Based on the results of the olive weights, it is clear that brown marmorated stink bugs do cause damage to olives. The insects were able to suck out enough fluid from the olives to cause noticeable weight differences between treatments. As each trial was carried out, brown marmorated stink bugs fed more on the olives causing a greater amount of fluid to be sucked out as each trial continued. From trial #1 to trial #4, the olives were more mature and the insects were more open to feeding on them which is a common characteristics of feeding behaviors of these agricultural pest (Venugopal et al., 2014). Brown marmorated stink bugs have been known to move between crops depending on availability, and more damage has been done to more mature fruits and vegetables (Venugopal et al., 2014). In addition to the olives losing weight, observable damage was also done to the olives. The insects were constantly feeding on the olives for the duration of the trials, which resulted in exterior and interior damage (2012, EPA). From feeding on the olives with their proboscis, a result was small black spots on the olive, later to be identified as fruit bleeding. More mature 29

36 olives that were fed on, resulted in discoloration, mold, and collapsing (Figure 19). Once each trial was over, olives were dissected under a microscope to observe any internal damage. Olives that were exposed to brown marmorated stink bug were discolored and rotten inside (Figure 19). Inside of olives also contained brown spots from where the proboscis pierced the fruit and as a result caused small amounts or corking which is common in stink bug damage (2012, EPA). Relative Humidity and Temperature: The relative humidity and temperature began to be recorded in trial #3 due to a high number of insects dying in the start of the trial in the treatments with the cotton swabs. When this data was compared to the data recorded in trial #4, it became clear that the humidity and temperature was not the cause of the quick deaths in trial #3. The relative humidity and temperature data were the same for both trials in corresponding treatments, but the insects in trial #4 did not die off as quick. The insects in trial #3 could have been older or been handled more roughly causing the mortality rate to increase. Overall, temperatures were the same throughout all of the treatments. The only change in temperatures were throughout the day and night. At night the temperatures would drop, but since the experiment was done in a controlled environment, the changes were never significant. The relative humidity shifted from treatment to treatment. The treatments with the highest relative humidity were the ones containing the saturated cotton swabs which were: Olives + Water + Insects, Control #2: Olives + Water, and Insects + Water. 30

37 Reproduction: Brown marmorated stink bugs were able to produce more egg masses and total number of eggs in some treatments than others. Olives + Insects + Water, produced the most consistent and large number of egg masses throughout all four trials. The Insects + Water treatment produced the next largest amount of egg masses. The fewest egg masses were produced in the treatment, Olives + Bugs. From this information we can conclude that with a water source, the insects are able to reproduce with olives or not. In the Olive + Bug treatment, in trial #2, no egg masses were produced for the entire trial when younger olives were used. However in trial #3, they were able to reproduce and make egg masses with the more mature olives. Egg mass production declined in all the treatments for trial #4, which was expected based on the amount of daylight these insects need to produce egg mass, which dropped below 14 hours a day (Smith, 1966, p ). (Figures 11, 12, 13, and 14). Feeding Behaviors: Based on digital imaging, it was found that these insects are more active and feed more in the afternoon hours of 2:00 PM 6:00 PM than they are in the morning from 7:00 AM 11:00 AM. This pattern can be explained by the temperature change in the room. In the morning it was colder, therefore the insects were not as active and did not feed as much (Wiman et al., 2014). Then in the afternoon with warmer temperatures, it caused the insects to be more active and that is when they were most likely to feed (Wiman et al. 2014).Probing of brown marmorated stink bugs in controlled environmental conditions has been done, and the results proved that if temperatures are too hot or cold, that is when feeding is at its minimum (Wiman et al. 2014). The ideal temperate when the most feeding takes place is between 16 and 17 degrees Celsius (Wiman et al. 2014). The treatment Olives + Insects had a higher feeding contact average than Olives + 31

38 Insects + Water. This was most likely because the Insects were relying more on the olives for food and water since there was no alternate source of water. (Figures 15, 16, 17, and 18). For the olive growers: For olive growers, they will have to develop more effective pest management strategies for combating the brown marmorated stink bug in the near future. Growers will have to apply insecticides early in the growing season since it has been found that brown marmorated stink bugs have higher death rates after the over-wintering period than the later generations (Leskey et al., 2013). The highest number of these insects in fields that have been infested, have been on the edge of the field (Venugopal et al., 2014)). It would be the most cost and environmentally beneficial to apply pesticides to the outer edge of crops (Venugopal et al., 2014). It also has been found that commonly used pesticides to combat brown marmorated stink bug are most effective during the 2 nd instar of these insects life cycle (Parker et al., 2015). Regular inspections of olive crops will have to be made if fields become infested with these stink bugs to apply pesticides in a timely manner for highest mortality rate possible. Within 12 days of pesticide applications during the 2 nd instar, there has been a 95% - 100% mortality rate (Parker et al., 2015). There are other methods that can be done that have been found to be more environmentally friendly. Essential oils have been tested on crops and traps to try and repel brown marmorated stink bugs (Zhang et al., 2013). This method was found to be highly effective and resulted in less crop damage and less insects found in stink bug traps (Zhang et al., 2013). However, this method is not a permanent fix since it is not killing or eradicating stink bug populations. More research is being done to help detect and manage these invasive stink bugs on a daily basis to help develop better pest management strategies for farmers would wide. 32

39 Table 1.Treatments and materials used in each of the trials. Treatments Trials Used Water Olives Insects Leaves Control #1: Olives 1, 2, 3, 4 X Control #2: Olives + Water 2, 3, 4 X X Insects + Water 2, 2, 4 X X Insects + Olives 1, 2, 3, 4 X X Insects + Olives + Water 1, 2, 3, 4 X X X Olive Leaves + Insects 3, 4 X X Olive Leaves + Insects + Olives 4 X X X 33

40 Figure 1. Distribution of brown marmorated stink bugs throughout California. 34

41 Figure 2 (A, B, C, D). A) Damage done by the Olive Fruit Fly that looks similar to brown marmorated stink bug damage. B) Brown marmorated stink bug damage on an avocado that is similar to the damage they did to olives. C) and D) Damage caused by brown marmorated stink bugs. 35

42 Number of BMSB alive 6 BMSB Surviorship Rate With Bugs + Olives Olives + Bugs Trial #1 Olives + Bugs Trial #2 Olives + Bugs Trial #3 Olives + Bugs Trial #4 Figure 3. Average survivorship rate of BMSB over increasing time (left to right) in the Bugs + Olives treatment for all trials. 36

43 Number of BMSB Alive 6 BMSB Surviorship Rate with Olives + Bugs + Water Olives + Bugs + Water Trial #1 Olives + Bugs + Water Trial #2 Olives + Bugs + Water Trial #3 Olives + Bugs + Water Trial #4 Figure 4.Average survivorship of BMSB over time (left to right) in the Olives + Bugs + Water treatment. 37

44 Number of BMSB alive 6 BMSB Surviorship Rate with Bugs + Water Bugs + Water Trial #2 Bugs + Water Trial #3 Bugs + Water Trial #4 Figure 5. Average survivorship rate of BMSB over increasing time (left to right) in the Bugs + Water treatment for all trials. 38

45 Number of BMSB alive 2.5 BMSB Surviorship Rate with Bugs + Olive Leaves Bugs + Leaves Trial #3 Bugs + Leaves Trail #4 Figure 6. Average survivorship rate of BMSB over increasing time (left to right) in the Bugs + Olive Leave treatment for all trials. 39

46 WEIGHT OF OLIVES (GRAMS) Average Weight Difference of Treatments in Trial # Olives Olives + Bugs Olives + Bugs+ Water 0 Olives + Water Figure 7. Compares the difference in weights (in grams of the olives used in each of the treatments in trial #1. 40

47 Average Weight Difference of Treatments in Trial # Olives Olives + Bugs Olives + Bugs+ Water Olives + Water Figure 8. Compares the difference in weights (in grams) of the olives used in each of the treatments in trial #2 41

48 Average Weight Difference of Treatments in Trial # Olives Olives + Bugs Olives + Bugs+ Water 0.67 Olives + Water Figure 9. Compares the difference in weights (in grams) of the olives used in each of the treatments in trial #3. 42

49 Average Weight Difference of Treatments in Trial # Olives Olives + Bugs Olives + Bugs+ Water 0.21 Olives + Water Figure 10. Compares the difference in weights (in grams) of the olives used in each of the treatments in trial #4. 43

50 NUMBER OF EGG MASSES (BLUE) AND NUMBER OF EGGS (ORANGE) Numbers of Egg Masses and Eggs in Olives + Bug Treatments Olives + Bugs Trial #1 Olives + Bugs Trial #2 Olives + Bugs Trial #3 Olives + Bugs Trial #4 0 Egg Masses Number of Eggs Figure 11. Comparison of the number of egg masses and eggs produced for each trial in the Olive + Insect treatments. 44

51 NUMBER OF EGG MASSES (BLUE) AND NUMBER OF EGGS (ORANGE) Number of Egg Masses and Eggs in Olives + Bugs + Water Treatments Olives + Bugs + WaterTrial #1 Olives + Bugs + Water Trial #2 Olives + Bugs + Water Trial # Olives + Bugs + Water Trial #4 Egg Masses Number of Eggs Figure 12. Comparison of the number of egg masses and eggs produced for each trial in the Olive + Insect + Water treatments. 45

52 NUMBER OF EGG MASSES (BLUE) AND NUMBER OF EGGS (ORANGE) Number of EggMasses and Eggs in Bugs + Water Treatments Bugs + WaterTrial #1 Bugs + Water Trial #2 Bugs + Water Trial #3 Bugs + Water Trial #4 53 Egg Masses Number of Eggs Figure 13. Comparison of the number of egg masses and eggs produced for each trial in the Insect + Water treatments 46

53 NUMBER OF EGG MASSES (BLUE) AND NUMBER OF EGGS (ORANGE) Number of Egg Masses and Eggs in Olive Leaves + Bugs Treatments Bugs + WaterTrial #1 Bugs + Water Trial #2 Bugs + Water Trial #3 Bugs + Water Trial #4 Egg Masses Number of Eggs Figure 14. Comparison of the number of egg masses and eggs produced for each trial in the Olives Leaves + Insects treatments. 47

54 INSECT CONTACTS PER HOUR Insect Contacts with Olives Per Hour: Trial # :00 AM 10:00 AM 2:00 PM 6:00 PM 6:00 AM 10:00 AM 2:00 PM 6:00 PM Trial #1 Insects + Olives + Water Trial #1 Olives + Insects (All Dead) Figure 15. Comparison of one hour time frames that brown marmorated stink bugs made contact with olives between two treatments (Insects + Olives + Water and Insects + Olives) in trial #1. 48

55 INSECT CONTACTS PER HOUR Insect Contacts with Olives Per Hour: Trial # :00 AM 10:00 AM 2:00 PM 6:00 PM 6:00 AM 10:00 AM 2:00 PM 6:00 PM Trial #2 Insects + Olives + Water Trial #2 Olives + Insects Figure 16. Comparison of one hour time frames that brown marmorated stink bugs made contact with olives between two treatments (Insects + Olives + Water and Insects + Olives) in trial #2. 49

56 INSECT CONTACTS PER HOUR Insect Contacts with Olives Per Hour: Trial # :00 AM 10:00 AM 2:00 PM 6:00 PM 6:00 AM 10:00 AM 2:00 PM 6:00 PM Trial #3 Insects + Olives + Water Trial #3 Olives + Insects Figure 17. Comparison of one hour time frames that brown marmorated stink bugs made contact with olives between two treatments (Insects + Olives + Water and Insects + Olives) in trial #3. 50

57 INSECT CONTACTS PER HOUR Insect Contacts with Olives Per Hour: Trial # :00 AM 10:00 AM 2:00 PM 6:00 PM 6:00 AM 10:00 AM 2:00 PM 6:00 PM Trial #4 Insects + Olives + Water Trial #4 Olives + Insects Figure 18. Comparison of one hour time frames that brown marmorated stink bugs made contact with olives between two treatments (Insects + Olives + Water and Insects + Olives) in trial #4. 51

58 Figure 19. Pitting, discoloration, and fruit bleeding damage to olive from brown marmorated stink bugs. 52