SPLAT Verb: A New Player in the Field of Semiochemical Control for Bark Beetles

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1 SPLAT Verb: A New Player in the Field of Semiochemical Control for Bark Beetles Christopher J. Fettig 1, Leif A. Mortenson 1, Robert A. Progar 1, A. Steven Munson 2, Beverly M. Bulaon 2, Daniel R. Cluck 2, Monica L. Gaylord 2, Sandra Kegley 2, Cynthia L. Snyder 2, Lia Spiegel 2, Brytten E. Steed 2, and Agenor Mafra-Neto 3 1 Research and Development, USDA Forest Service 2 Forest Health Protection, USDA Forest Service 3 ISCA Technologies Inc. California Forest Pest Council McClellan, CA, 2015

2 Specialized Pheromone & Lure Application Technology A matrix-type diffusion controlled-release device. As such, the release of active ingredients is determined by Fick s First Law of Diffusion, which states that molecules move from regions of high concentration to regions of low concentration at a rate that is directly proportional to its concentration gradient. Biodegradable. Inerts have been determined by USEPA to be suitable for food use. SPLAT products are certified for organic use.

3 C. Fettig, USDA Forest Service Verbenone (4,6,6-trimethylbicyclo[3.1.1]hept-3-en-2-one) Principle antiaggregant of several bark beetles. Produced in small amounts by autoxidation of the monoterpene α-pinene, but primarily through metabolic conversion by bark beetles of inhaled and ingested α-pinene to cis- and trans-verbenol, which are metabolized by yeasts in the alimentary system and beetle galleries to verbenone (Progar et al. 2014). Inhibits additional beetles from infesting the target tree, presumably limiting the number of beetles to a density that increases the likelihood of brood survival. A 5-g pouch was first registered by the U.S. Environmental Protection Agency (EPA) in 1999 for southern pine beetle.

4

5 Mountain Pine Beetle (MPB) Throughout BC, AB, most of the western U.S., and northern Mexico. Primary hosts are pines, most notably lodgepole, ponderosa, sugar, and whitebark. Recent outbreaks have been severe, long-lasting and well-documented. >27 million ha affected; >11 smallest U.S. states ( SC) combined. C. Fettig, USDA Forest Service

6 C. Fettig, USDA Forest Service Prototype 1 Individual Tree (Lodgepole, WY, 2011) 21 PICO treated with SPLAT formulated with ( )-verbenone by applying four equally-sized dollops to the tree bole at ~3 m in height with a pneumatic system. 15 PICO (26.2 ± 1.3 cm) received 32 g of ( )-verbenone tree 1 (533 g of SPLAT Verb tree 1 ) 6 PICO (26.4 ± 2.3 cm) received 39.3 g of ( )-verbenone tree 1 (655 g of SPLAT Verb tree 1 ). 30 PICO (32.0 ± 0.8 cm) served as untreated controls. SPLAT is biologically inert and biodegradable. All trees baited with exo-brevicomin and trans-verbenol.

7 A. Mafra-Neto, ISCA Technologies Inc.

8 Prototype 1 Individual Tree (Lodgepole, MT, 2011) B. Steed, USDA Forest Service

9 A. Mafra-Neto, ISCA Technologies Inc. Prototype 2 Individual Tree (Lodgepole, WY, 2012) 30 PICO (27.1 ± 0.8 cm) treated with SPLAT Verb applied at 7.0 g of ( )-verbenone/tree (70 g of SPLAT Verb/tree) as 4 ~17.5-g dollops (~5.5 cm diam. X 1.2 cm ht.) to the tree bole (at cardinal directions) with a caulking gun. 30 PICO (32.3 ± 1.2 cm) served as untreated controls.

10 Prototype 2 Individual Tree (Lodgepole, MT, 2012) Fettig, C.J. et al J Econ Entomol 108:

11 SPLAT Verb Individual Tree (Lodgepole, MT, 2013) Fettig, C.J. et al Crop Protection, in review. C. Fettig, USDA Forest Service

12 Prototype 2 Area Wide (Lodgepole, ID, 2012) SPLAT Verb was applied at g of ( )-verbenone/plot to tree boles (both host and nonhosts) at ~2 m in height on the northern aspect in appropriate dollop sizes (~17.5 g) to achieve adequate (~5 6 m, depending on tree distributions) coverage. 7-g ( )-verbenone pouches stapled to the tree bole at ~2 m in height on the northern aspect along a ~9.1 by 9.1-m grid (125 U/ha; g of ( )- verbenone/plot) Untreated control. C. Fettig, USDA Forest Service

13 Prototype 2 Area Wide (Lodgepole, ID, 2012) Fettig, C.J. et al J Econ Entomol 108:

14 Fettig, C.J. et al J Econ Entomol 108:

15 C. Fettig, USDA Forest Service SPLAT Verb Zone of Inhibition (Lodgepole, UT, 2013) Seventy g of SPLAT Verb [7 g of ( )- verbenone] were applied as four ~17.5-g dollops at ~2.5 m in height to the bole of one PICO (mean dbh ± SEM = 36.7 ± 1.2 cm) at the center of each of eight blocks. One 16-unit multiple-funnel trap baited with a commercially-available trap lure was placed at 1, 2, 4, 8 and 16 m from the SPLAT Verb-treated tree at a randomly-selected bearing of 0, 72, 144, 216 or 288º.

16 There was no significant distance X gender interaction (P = 0.46), and therefore results relate equally to both male and female responses. This is of practical importance as females initiate host selection and therefore inhibition of females is critical to preventing colonization and tree mortality. No differences were observed among captures at 1, 2 or 4 m from the point of release of SPLAT Verb, but significantly fewer MPB were collected at 1 and 2 m compared to 8 m. Fettig, C.J. et al J Econ Entomol 108:

17 Fettig, C.J. et al J Econ Entomol 108: Individual Tree (Lodgepole, WY, 2012) Significantly fewer PICO (percentage of trees) were killed on ha circular plots surrounding PICO treated with SPLAT Verb compared to the untreated control, suggesting attraction was disrupted at levels sufficient to cause tree protection within 11 m of the point of release of SPLAT Verb. B. Steed, USDA Forest Service

18 SPLAT Verb - Chemical Composition 17.5 g dollops were analyzed by gas chromatography (GC) to estimate release rates, and GC and gas chromatography mass spectrometry (GC-MS) to determine chemical identity and composition. Average release during a 30-d period was 28.6 mg/d. Assuming release plateaus at ~15 mg/d (i.e., mean release was 14.7 mg/d for days 10 30) and continues at this rate the reservoir of verbenone would be sufficient to cover most of the flight activity period. Only after dollops were aged for >12 mo were trace amounts of chrysanthenone found. Fettig, C.J. et al J Econ Entomol 108: C. Fettig, USDA Forest Service

19 SPLAT Verb Individual Tree (Lodgepole, MT, 2014) Fettig, C.J. et al. unpubl.

20 SPLAT Verb Area Wide (Lodgepole, MT, 2014) B. Steed, USDA Forest Service

21 SPLAT Verb Diffuse/Chemical Composition (7.5 g, 12.5 g, 17.5 g, 35 three elevations, 2015) Gas chromatography to estimate release rates, and GC-MS to determine chemical identity and composition R. Progar, USDA Forest Service

22 SPLAT Verb Individual Tree (Sugar pine, CA, 2014) Fettig, C.J. et al Crop Protection, in review.

23 Rim Fire: treated 86 trees (121.9 ± 4.3 cm dbh, max = cm) w/ 10.0 g a.i.

24 SPLAT Verb Five-needle Pines (Whitebark, CA, OR and MT, 2015) (SWWP, AZ, 2015) R. Progar, USDA Forest Service R. Progar, USDA Forest Service C. Fettig, USDA Forest Service

25 SPLAT Verb Plus (Ponderosa, CA and OR, 2014) Fettig, C.J. et al. unpubl.

26 Christopher J. Fettig Fettig, C.J., A.S. Munson, M. Reinke, and A. Mafra-Neto A novel semiochemical tool for protecting lodgepole pine from mortality attributed to mountain pine beetle (Coleoptera: Curculionidae). J Econ Entomol 108: Fettig, C.J., B.E. Steed, B.M. Bulaon, R.A. Progar, A.S. Munson, and A. Mafra-Neto The efficacy of SPLAT Verb for protecting individual Pinus contorta, Pinus ponderosa, and Pinus lambertiana from colonization by Dendroctonus ponderosae. Crop Protection, in review. Mafra-Neto, A., F.M. de Lame, C.J. Fettig et al Manipulation of insect behavior with Specialized Pheromone & Lure Application Technology (SPLAT ). Natural Products for Pest Management. ACS Publications, Washington, p Mafra-Neto, A., C.J. Fettig, A.S. Munson, and L.L. Stelinski The use of repellents formulated in Specialized Pheromone and Lure Application Technology (SPLAT ) for effective insect pest management. Insect Repellents Handbook, Second Edition. Taylor & Francis, Boca Raton, p C. Fettig, USDA Forest Service Mafra-Neto, A., C.J. Fettig, A.S. Munson, et al Development of specialized pheromone and lure application technologies (SPLAT ) for management of coleopteran pests in agricultural and forest systems. Biopesticides: State of the Art and Future Opportunities. ACS Publications, Washington, p

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