1. Please state project objectives and what work was done this quarter to address them: With many of the stated objectives completed for this project, we have continued to work on Objectives 1a, 1b, 1e, and 2c. 1a: Develop monitoring methods to time management actionsWe have been working with the Stelinski lab to identify plant-derived odors that are attractive to lebbeck mealybug crawlers. In the previous research update, we listed the mechanically induced volatiles of citrus Table 1. So, we conducted behavioral response bioassay to describe the response of hibiscus mealybugs to two doses of the induced citrus volatiles using a Y-tube olfactometer (Fig. 1).We recorded the response of 2nd to 3rd instar immatures of hibiscus mealybugs to two dilution levels (0.1 µg/µL and 0.01 µg/µL) of the stock solution (1µg/µL) of each volatile.Results of behavioral response showed that hibiscus mealybugs were significantly attracted to scent of synthetic ß-Ocimene, .-Terpinene, Farnesene, and Citronellal in the olfactometer at both dilutions. While the mealybugs were significantly repelled by scents of a-Humulene, ß-Elemene, and ß-Caryophyllene at both dilution levels (Fig. 2 and 3). At 0.1 µg/µL, Limonene significantly repelled hibiscus mealybugs but mealybug repulsion was not significant at 0.01 µg/µL (Fig. 2 and 3). Greater repulsion was recorded in mealybugs exposed to ß-Pinene and d-3-Carene at 0.1 µg/µL, but significant attraction was recorded when the mealybugs were exposed to both volatiles at a lower concentration i.e., 0.01 µg/µL (Fig. 2 and 3). Similar contradictory results were also recorded in mealybugs exposed to Citronellyl acetate (Fig. 2 and 3). Our results suggest that ß-Ocimene, .-Terpinene, Farnesene, and Citronellal can potentially be used as attractants in lures for monitoring or mass trapping of hibiscus mealybugs in the field. While a-Humulene, ß-Elemene, and ß-Caryophyllene, are potential repellent volatiles of hibiscus mealybug. 1b: Expand laboratory insecticide and adjuvant screeningWe have obtained several chemistries used for management of other mealybug in other cropping systems and are currently evaluating their efficacy against lebbeck mealybug in lab assays. Any that look promising will be taken to the grove for testing this fall. These studies are currently underway therefore there are currently no data to report. 1e: Evaluate management options for IPCsWe have tested several options for cleaning IPCs for reuse. To date, the most effective clean up tool is 1-5% dish soap (we used Dawn) in water. IPCs should be fully immersed, agitated, and left to soak for approximately 24 hours. After soaking, a thorough rinsing should be performed to remove residues. This method should not be used in groves as dish soap can cause phytotoxicity to leaves and fruit. 2c. Determine what insecticide chemistries inhibit feedingWhile this objective has been waylaid as we await the wire necessary to fully document feeding in order to dig into feeding inhibition, we have made some progress troubleshooting our challenges as described below. In our preliminary EPG recordings, we observed two main issues:1) Noise problem: preventing us from distinguishing E1 (phloem salivation) from E2 (phloem ingestion). Initiative to fix the issue: We installed two voltage regulators (Furman®, Model: P-1800 AR) on the two EPG monitors and observed an improvement in the signal quality.1) Behavior problem: Most of the recordings are spent in the pathway phase (test probing) and little time is spent on phloem or xylem ingestion (<15%). Yet, mealybugs are phloem feeders and should spend more time feeding on the sap elements (xylem and/or phloem). Initiative to fix the issue: We hypothesize that the gold wire is too thick (25µm diameter) compared to the mealybug, preventing it from expressing its natural feeding behavior. We ordered some platinum Wollaston (2.5µm diameter) via the company Sigmund Cohn Inc. in March. The wire should arrive by mid-July or beginning of August. We also received a thinner gold wire (13µm diameter) from a collaborator. Within the next months we are planning to test the two wires and see if we observe a change in probing behavior. We believe that these wires will be more flexible and allow better movement of the mealybug.We are also trying to build a waveform library to better interpreter the biological meaning of the waveforms observed. Previously, we calculated the inherent resistance (Ra) of the mealybug and found Ra=1012. We then selected three input resistance (Ri) that bracket the Ra of N. viridis:- 109 Ri with substrate voltage at 250mV (some R component)- 1010 Ri with substrate voltage at 100mV- 1013 Ri with substrate voltage at 0mV (pure emf signal)We will continue to record at these three different Ri and compare the waveform's appearance according to the settings. Based on our results, we will make recommendation about the best monitor setting and the best tethering method for future studies with mealybugs. You will find a summary of the waveforms identified in previous recordings and their characteristics in Table 1. 2. Please state what work is anticipated for next quarter: We will continue work on the objectives listed above. 3. Please state budget status (underspend or overspend, and why): We reduced staff assigned to this project for objective for Obj. 2c as we had to order the new wire to complete the work. The wire should be here by the end of July and work will resume once it is here. Staffing assignments for the upcoming quarter reflects this expectation. 4. Please show all potential commercialization products resulting from this research, and the status of each:If we are able to develop an attractive lure, that would be a product that can be commercialized but that end point will be outside the timeline of the funding of this project.