TY - JOUR
AU1 - Elmquist, Dane C
AU2 - Landolt, Peter J
AU3 - Cooper, William Rodney
AU4 - Reed, Hal
AU5 - Foutz, Jillian
AU6 - Clepper, Timothy
AU7 - Kacprzyk, Bryon
AU8 - Teig, Donald
AU9 - Zack, Richard S
AB - Abstract Polistes paper wasps in the Fuscopolistes subgenus (Hymenoptera: Vespidae) can be serious pests when they swarm at tall man-made structures. Chemical attractants may be useful to trap such paper wasps when they achieve pest status. Polistes venom has been shown to elicit a variety of behavioral responses in congeneric wasps, making it a source for potential chemical attractants. The compound N-(3-methylbutyl)acetamide is a principal volatile component in the venom of many female vespid wasps, including numerous Polistes species. We report the presence of N-(3-methylbutyl)acetamide in autumn gynes of Polistes metricus Say, Polistes bellicosus Cresson, and Polistes dorsalis (F.), as well as workers of Polistes aurifer (Saussure), P. bellicosus, P. metricus, and P. dorsalis. In field tests conducted in Florida, Georgia, South Carolina, and Washington, N-(3-methylbutyl)acetamide attracted male and female P. aurifer and P. metricus, as well as male P. dorsalis and P. bellicosus. Thus, N-(3-methylbutyl)acetamide may be a useful lure for trapping these paper wasps in pest situations. Paper wasps, Polistes, venom, attractant, N-(3-methylbutyl)acetamide Polistes paper wasps (Hymenoptera: Vespidae) are beneficial predators of pest arthropods (Gould and Jeanne 1984), but several species of Polistes are pestiferous in the southern United States when they swarm at tall man-made structures (Reed and Landolt 1991). These wasps are primarily Polistes fuscatus (F.), Polistes metricus Say, Polistes bellicosus Cresson, and Polistes dorsalis (F.), all within the subgenus Fuscopolistes of Richards (1978). The autumn swarming of new reproductive females (gynes) and males of paper wasps at towers and tall buildings appears to involve gynes seeking sheltered overwintering sites and males attempting to intercept those gynes as potential mates (Reed and Landolt 1991). These activities of paper wasps at swarming and overwintering sites suggest several pheromone-mediated behaviors that might be manipulated to mitigate the wasp’s pest status. More recently, similar behavior of Polistes males and females has been observed at air traffic control towers on Air Force Bases in South Carolina, Georgia, and Florida (unpublished observations by U.S.A.F. personnel and P.J. Landolt). Studying the chemical ecology and swarming behavior of reproductive Polistes wasps could yield attractants such as sex and aggregation pheromones (Post and Jeanne 1983, 1984; Reed and Landolt 1990; MacKenzie et al. 2008; Elmquist et al. 2018) that might be useful as trap lures or baits to mitigate pest populations. Post and Jeanne (1983, 1984), using laboratory assays, first showed attraction of P. fuscatus males to females and to extracts of tagmata and glands of females. Results of those assays indicated that female venom glands are the source of an attractant for males of both P. fuscatus and Polistes exclamans Vierick. Venom of female Polistes is a chemically complex, multifunctional blend of peptides and proteins as well as several relatively volatile compounds (Turillazzi 2006). Vespid wasp venom contains allomones that are inducers of pain, inflammation, tissue damage, and other physiological effects (Habermann 1971). Vespid wasp venom is also the source of semiochemicals that have defensive and communicative functions, such as alarm pheromones demonstrated in multiple genera (Bruschini et al. 2006a; Cheng et al. 2017; Heath and Landolt 1988; Jeanne 1981, 1982; Landolt and Heath 1987; Thiery et al. 2018; Veith et al. 1984). Also, venom of vespid social parasites may include dominance allomones (Jeanne 1977, Landolt and Akre 1979, Bruschini and Cervo 2011, Reed and Akre 1982). Although vespid wasp venom is well known as delivered through the sting directly into animal tissue, venom is also dispensed by wasps with a spraying action (Vespula germanica [F.], Maschwitz 1964; Dolichovespula arenaria [F.], Greene et al. 1976), or the emission of small amounts of venom with extrusion of the stinger during courting or mating (West-Eberhard 1969, Romani et al. 2005, Post and Jeanne 1983, Reed and Landolt 1990, Turillazzi 2006). Grooming motions of the legs spreads droplets of venom from the tip of the sting onto the wasp’s cuticle (Post and Jeanne 1984, Baracchi et al. 2011). Such grooming actions could be allowing volatile chemicals within the venom to evaporate from a larger surface area and function as pheromones or allomones, or other semiochemicals. The chemical N-(3-methylbutyl)acetamide (MBA) has been reported as a principal volatile component of the venom of a number of vespine social wasps; V. germanica (Aldiss 1983; Weston et al. 1997 reported as N-isopentylacetamide), Vespula squamosa (Drury) (Heath and Landolt 1988), Vespula maculifrons (Buysson) (Landolt et al. 1995), and Dolichovespula maculata (L.) (Jimenez et al. 2016). The compound was shown to have an alarm pheromone function in eliciting recruitment and attack behavior by V. squamosa (Heath and Landolt 1988) and V. maculifrons at nest sites (Landolt et al. 1995). MBA has also been found in the venom of Polistes dominula Christ, Polistes gallicus (L.), Polistes nimpha Christ, Polistes sulcifer Zimmerman, and Polistes olivaceus DeGeer (Bruschini et al. 2006b) and the epiponines (Hymenoptera: Vespidae) Polybia occidentalis and Polybia sericea (Olivier) (Dani et al. 2000). Given the presence of MBA in the venom of numerous vespids, the attraction of male Polistes to the venom of both conspecific and heterospecific females (Post and Jeanne 1983, 1984), and the multispecies nature of Fuscopolistes mating swarms (Reed and Landolt 1991), we hypothesize that this chemical may have multiple functions, depending on species, caste, sex, and context, including as an attractant that plays a role in forming and maintaining mating swarms observed at towers. For this reason, we sought to determine the presence and amount of MBA in the sting apparatus of both worker and fall gynes of Polistes paper wasps found in swarms at towers in the southeastern United States (Reed and Landolt 1991). We also field-tested MBA to determine its attractiveness to Fuscopolistes gynes, both at swarm sites at U.S. Air Force air traffic control towers in the southern United States and at P. aurifer (Fuscopolistes) male patrol routes in a rural area of Yakima County, Washington. Studying the chemical basis of male and female Polistes behavior during mating aggregations will improve our understanding of paper wasp mating strategies and suggest semiochemical-based techniques for pest management. Methods and Materials Collection and Handling of Wasps Polistes bellicosus and P. metricus workers were collected in July, and gynes were collected in October 2018 from Pearl River County, Mississippi. Polistes dorsalis workers were collected in July, and gynes were collected in October/November 2018 from Alachua County, Florida. Polistes aurifer workers were collected at the USDA-ARS experimental farm east of Moxee, Yakima Co., Washington in July 2018. Southeastern U.S. paper wasp species identifications were confirmed using Krispyn and Hermann (1977) and Buck et al. (2008). Voucher specimens of all species described in these studies are deposited in the Maurice T. James Entomological Collection at Washington State University, Pullman, WA. Wasps caught in the field were placed in plastic snap cap vials and kept in a cooler to reduce activity. Wasps caught in Mississippi and Florida were then stored in a laboratory refrigerator for 1–3 d before 1-d shipping to the USDA-ARS Temperate Tree Fruit and Vegetable Research Laboratory near Wapato, Yakima County, WA (Yakima laboratory). Polistes aurifer field-collected in Washington were handled as described above, and transported within several hours to the laboratory. All wasp species and castes were housed separately in aluminum frame and stainless-steel screen cages (21 × 21 × 21 cm) or BugDorm plastic screen cages (30.5 × 29 × 31 cm) (BioQuip Products Inc., Rancho Dominguez, CA). Cages of wasps were further contained in insect rearing tents (60 × 60 × 60 cm) (BioQuip). Cages of wasps were placed in a room with a controlled environment at 25.5 ± 2.0°C under a natural light regimen (14L:10D) with 35–40% RH. Wasps were provided water and a sugar water solution (17 % wt/vol; Signature Kitchens fine granulated cane sugar, Albertsons Co., Boise, ID) on six to eight 2-cm diameter cotton balls placed in the bottom of each 8.5 cm diameter plastic Petri dish. Water and sugar water were replenished daily. Deceased wasps were removed from the cages. Individuals that were on the sides or tops of the screen cages were selected to dissect for MBA quantification. MBA Quantification Five P. metricus workers were placed in a freezer (−8°C for 15 min) before the sting apparatuses were dissected using stainless-steel #4 dissecting forceps. We considered the sting apparatus as the stinger (consisting of a dorsal sheath and two ventral lancets), the sting bulb, the venom reservoir, and the acid and alkaline glands (venom and Dufour’s glands) (Duncan 1939, Landolt and Akre 1979). After dissection, the sting apparatus from a single individual was immediately placed in 100 μl of dichloromethane (DCM) in a 2.5 ml glass vial with a conical bottom and ground with a glass stir stick. After grinding, the sting apparatus remained in the vial in DCM for 30 min. After this extraction period, the sting apparatus extract solution was pipetted into a 4 ml glass vial and reduced to 10 μl under a flow of nitrogen gas. Two microliters of the 10 μl of extract were manually injected into a gas chromatograph with flame ionization detection, and a coupled gas chromatograph mass spectrometer (GC-MS) (see below). This process was replicated five times to provide quantitation of MBA in the venom of five wasps for each species and caste of wasp studied. Since workers of Polistes species are capable of mating we determined the mating status of the wasps we used for venom analyses as the presence or absence of sperm in the spermathecae. Only workers that were unmated (lacked sperm in spermathecae) were used for worker venom MBA quantification. After the sting apparatus was dissected, the spermatheca of each wasp was removed and dry mounted to a microscope slide and stained with Geimsa stain (Procedure NO. GS-10, Sigma Aldrich, St. Louis, MO). Stained spermathecae were analyzed under 200× magnification using a Zeiss Axiolab A.1 phase-contrast compound microscope (Zeiss United States, San Diego, CA). Stained spermathecae from dissected wasps were compared to a spermatheca containing visible sperm from a mated gyne as a positive control. Quantification of MBA in gynes was limited to female wasps in which we could clearly assess the presence of sperm in the spermatheca. Venom extracts and chemical standards were analyzed in splitless mode using an HP 6890 GC with flame ionization detection, fitted with a DB-5 capillary column (30 m × 0.25 mm ID, 0.25 µm film thickness) (Agilent, Santa Clara, CA). Helium was the carrier gas at 1.3 ml/min constant flow. The GC oven temperature was programmed for 5 min at 40°C, increased 15°C/min to 140°C and then held for 1 min, then increased 20°C/min to 250°C, and then held for 5 min. Injector and detector temperatures were set at 250°C. Identification of MBA in the sting apparatus extract was based on comparison of the retention times to an authentic MBA synthetic standard (Oakwood Chemical Co., Estill, SC). The sting apparatus extracts analyzed contained 25 ng/μl of undecane as an internal standard. Extract injections of 2 μl were made manually using 10 μl syringes (Agilent). GC-MS was also used to confirm and quantify the MBA present in the sting apparatus extract. GC-MS analysis of extracts was conducted utilizing a HP 5973 MS coupled with a HP 6890 GC equipped with a Rxi-5MS capillary column (30 m × 0.25 mm ID, 0.25 μm film thickness) (Restek Corporation, Bellefonte, PA). Helium was the carrier gas at 1.3 ml/min constant flow. The GC oven temperature was programmed for 5 min at 40°C, 15°C/min increase to 150°C, 25°C/min increase to 300°C, and then held for 5 min. Injector temperature was set at 250°C. The MS transfer-line was set at 280°C, the MS source at 230°C, and the MS quad at 150°C. Mass spectra were taken in EI mode at 70.0eV in the range from 40 m/z to 260 m/z with a scanning rate of 6.10 scan/s and 6 min solvent delay. GC-MS data were processed with the MSD-Chemstation software ver. E.02.02.1431 (Agilent). Extract injections of 2 μl were made manually in splitless mode. Identification of MBA in the sting apparatus and gland extracts was based on comparison of retention times and mass spectrum with those of an authentic MBA synthetic standard. One microliter of a 25 ng/μl solution of synthetic MBA in DCM was injected prior to the sting apparatus analysis series for each species and caste to ensure that the retention time was consistent in each run for the entire analysis series. The amount (ng) of MBA per sting apparatus was quantified using a five-point standard calibration curve prepared with standard solutions of MBA at 0, 25, 50, 100, and 250 ng/μl MBA (r2 = 0.99). MBA in each sample was characterized using one target quantifier ion and qualifier ions to establish a method to automate quantification of the MBA peak area in each individual extract sample relative to the standard curve using MSD-Chemstation (Agilent). Amounts of MBA in workers and gynes were compared between species where both castes were analyzed using a two-sample t-test in R version 1.0.143 (R Core Team 2017). Field Test of Attractiveness of an MBA Lure Determining paper wasp attraction to MBA in the field involved deploying pairs of sticky traps (TrapStik for wasps, mud daubers, and carpenter bees, Sterling International Inc., Spokane, WA); replicates consisted of one trap baited with an MBA lure paired with a control trap at multiple sites in the states of WA, OK, FL, GA, and SC. Traps were maintained at each location in each state in October, November, or December of 2018 (Table 1). A total of 91 replicate pairs were placed across seven different locations. Trap locations were selected due to their potential as male swarming/patrolling locations or female aggregation/overwintering locations for Polistes (Fuscopolistes) species. Replicate pairs of traps were placed at least 30 m apart (Landolt et al. 1999) at all sites with one exception. Due to space limitations at the trapping location in OK, replicate pairs were placed 15 m apart. Traps and lures were replaced every 6 or 7 d. Traps were removed to a laboratory and placed in a freezer until paper wasps caught on each trap were identified and quantified by species and sex. Table 1. Locations and dates of field bioassays with MBA-baited and control traps throughout the United States in autumn 2018 Location . Trapping dates . Ahtanum State Forest, Yakima Co., WA 4–11 Oct., 11–17 Oct. USDA-ARS Farm, Yakima Co., WA 24–30 Sept., 18–25 Oct. Central WA Agricultural Museum, Yakima Co., WA 3–10 Oct., 11–17 Oct., 18–25 Oct. Eglin AFB, Okaloosa Co., FL 20–27 Nov. Shaw AFB, Sumter Co., SC 28 Nov. –5 Dec. Moody AFB, Lowndes Co., GA 29 Nov.–4 Dec. Oral Roberts University, Tulsa Co., OK 11–18 Oct., 23–30 Oct. Location . Trapping dates . Ahtanum State Forest, Yakima Co., WA 4–11 Oct., 11–17 Oct. USDA-ARS Farm, Yakima Co., WA 24–30 Sept., 18–25 Oct. Central WA Agricultural Museum, Yakima Co., WA 3–10 Oct., 11–17 Oct., 18–25 Oct. Eglin AFB, Okaloosa Co., FL 20–27 Nov. Shaw AFB, Sumter Co., SC 28 Nov. –5 Dec. Moody AFB, Lowndes Co., GA 29 Nov.–4 Dec. Oral Roberts University, Tulsa Co., OK 11–18 Oct., 23–30 Oct. Open in new tab Table 1. Locations and dates of field bioassays with MBA-baited and control traps throughout the United States in autumn 2018 Location . Trapping dates . Ahtanum State Forest, Yakima Co., WA 4–11 Oct., 11–17 Oct. USDA-ARS Farm, Yakima Co., WA 24–30 Sept., 18–25 Oct. Central WA Agricultural Museum, Yakima Co., WA 3–10 Oct., 11–17 Oct., 18–25 Oct. Eglin AFB, Okaloosa Co., FL 20–27 Nov. Shaw AFB, Sumter Co., SC 28 Nov. –5 Dec. Moody AFB, Lowndes Co., GA 29 Nov.–4 Dec. Oral Roberts University, Tulsa Co., OK 11–18 Oct., 23–30 Oct. Location . Trapping dates . Ahtanum State Forest, Yakima Co., WA 4–11 Oct., 11–17 Oct. USDA-ARS Farm, Yakima Co., WA 24–30 Sept., 18–25 Oct. Central WA Agricultural Museum, Yakima Co., WA 3–10 Oct., 11–17 Oct., 18–25 Oct. Eglin AFB, Okaloosa Co., FL 20–27 Nov. Shaw AFB, Sumter Co., SC 28 Nov. –5 Dec. Moody AFB, Lowndes Co., GA 29 Nov.–4 Dec. Oral Roberts University, Tulsa Co., OK 11–18 Oct., 23–30 Oct. Open in new tab Traps were paper-card cylinders (9.7 × 11.2 × 29.2 cm) colored with green and yellow hexagons and coated with an external adhesive. Lures consisted of 200 μl of a stock solution of 50 mg MBA/20 ml in isopropanol loaded onto a 2.5 cm cotton wick inside of a 4 ml polypropylene vial for a dose of 500 µg MBA. A 3 mm hole was drilled in the cap of the vial to release volatilized MBA. After loading, lures were placed in a laboratory fume hood for 24 h prior to placement in the field. Two hundred microliters of isopropanol on a cotton wick in a 4 ml polypropylene vial served as a control. MBA lures were placed directly on the adhesive surface near the top portion of the trap. The vial cap was positioned with the hole in the lid directed upwards. Vials were further secured to the traps using 26-gauge wire which was wrapped around the vial and trap. Control dispensers were attached to their respective traps in the same way. Ten replicate trap pairs were placed in three locations in Yakima Co., WA and maintained for 2–3 wk depending on the site. In Yakima Co, traps were hung on shepherd hooks (Lowes Company Inc., Mooresville, NC) 65 cm above the ground. Ten replicate trap pairs were hung on the roof of a dormitory at Oral Roberts University in Tulsa, OK and maintained over 2 wk. Four replicate trap pairs were tested in Okaloosa Co., FL and in Lowndes Co., GA, respectively and maintained for 1 wk. Three replicate trap pairs were tested in Sumter Co., SC and maintained for 1 wk. All traps at sites in FL, GA, and SC were placed near the top of towers at U.S. Air Force bases (AFB). Numbers of P. aurifer males and females caught on treated and control traps at the three locations in Yakima County, Washington were quantified. Lure response data (number of wasps caught on respective traps) did not conform to assumptions of normality and equal variance. We used negative binomial regression with a log link to analyze numbers of wasps trapped. The model was fit by maximum likelihood with Laplace approximation (Bolker et al. 2009). Lure type (MBA or control) was modeled as a fixed effect with replicate trap pairs nested in location as a random effect. For tower sites in FL, GA, and SC, numbers of P. metricus males and females, P. bellicosus males, and P. dorsalis males caught on baited and control traps were quantified and data from all these sites were pooled and subject to the same analyses as above. Numbers of P. bellicosus and P. dorsalis females trapped were too few (<10 per site) for statistical analyses. Numbers of male and female P. fuscatus and male P. metricus wasps caught on traps in OK were not pooled with the rest of the tower data due to the unavoidable deviation in distance between replicate pairs. Numbers of female P. metricus were too few (<10) at this site for statistical analyses. Wasps caught on each MBA-baited and control trap at this single location were quantified and compared using a two-sample t-test. Finally, we pooled all of the wasps trapped across all locations where replicate pairs were at least 30 m apart (81 replicate pairs across 6 locations) to compare total numbers caught on each treatment across four different states (WA, FL, GA, and SC). The total wasp numbers captured on baited versus control traps were compared as described above. All statistical tests were conducted using R version 1.0.143 (R Core Team 2017) using the lme4 package in R (Bates et al. 2015). Results MBA Analysis MBA was present in the sting apparatus extract of every wasp from each species and caste sampled (Table 2). Differences between workers and fall gynes were evaluated for P. metricus, P. bellicosus, and P. dorsalis. In P. metricus, amounts of MBA present in workers compared to fall gynes were similar (t8 = 0.60, P = 0.56). The sting apparatus of P. dorsalis workers contained significantly more MBA compared to P. dorsalis fall gynes (t8 = 3.41, P = 0.009). Extracts of the sting apparatus of P. bellicosus workers contained MBA in amounts that were numerically greater on average but were not statistically different compared to amounts in fall gynes (t8 = 1.89, P = 0.09). Table 2. MBA (ng/wasp sting apparatus) in Polistes Wasp . Caste . Mean ± SEM . P. aurifer Worker 32.3 ± 7.3 P. bellicosus Worker 115.0 ± 29.8 Fall gyne 54.3 ± 12.0 P. dorsalis* Worker 174.0 ± 28.4 Fall gyne 67.0 ± 13.3 P. metricus Worker 64.8 ± 14.4 Fall gyne 75.1 ± 9.4 Wasp . Caste . Mean ± SEM . P. aurifer Worker 32.3 ± 7.3 P. bellicosus Worker 115.0 ± 29.8 Fall gyne 54.3 ± 12.0 P. dorsalis* Worker 174.0 ± 28.4 Fall gyne 67.0 ± 13.3 P. metricus Worker 64.8 ± 14.4 Fall gyne 75.1 ± 9.4 * indicates a significant difference in MBA ng/wasp between conspecific castes at P < 0.05. Open in new tab Table 2. MBA (ng/wasp sting apparatus) in Polistes Wasp . Caste . Mean ± SEM . P. aurifer Worker 32.3 ± 7.3 P. bellicosus Worker 115.0 ± 29.8 Fall gyne 54.3 ± 12.0 P. dorsalis* Worker 174.0 ± 28.4 Fall gyne 67.0 ± 13.3 P. metricus Worker 64.8 ± 14.4 Fall gyne 75.1 ± 9.4 Wasp . Caste . Mean ± SEM . P. aurifer Worker 32.3 ± 7.3 P. bellicosus Worker 115.0 ± 29.8 Fall gyne 54.3 ± 12.0 P. dorsalis* Worker 174.0 ± 28.4 Fall gyne 67.0 ± 13.3 P. metricus Worker 64.8 ± 14.4 Fall gyne 75.1 ± 9.4 * indicates a significant difference in MBA ng/wasp between conspecific castes at P < 0.05. Open in new tab Field Test of Attractiveness of an MBA Lure Male and female P. aurifer were caught at all Washington trapping locations (Table 3). Trap catch results indicated that male and female P. aurifer were more likely to be caught on traps baited with MBA compared to control traps (male χ 2 = 23.15, P < 0.001; female χ 2 = 8.31, P = 0.004). Three Polistes (Fuscopolistes) species were caught at all tower locations in the southeastern United States (P. metricus, P. dorsalis, P. bellicosus; Table 3). Across all tower locations in GA, SC, and FL, male P. metricus were more likely to be caught on MBA-baited traps than control traps (Wald χ 2 = 24.34, P < 0.001). Female P. metricus were also caught in higher numbers on MBA-baited traps than controls (Wald χ 2 = 9.31, P = 0.002). The same trend was true for P. bellicosus males (Wald χ 2 = 28.72, P < 0.001) and P. dorsalis males (Wald χ 2 = 20.69, P < 0.001) on towers in GA, FL, and SC. Polistes metricus males and P. fuscatus males and females were trapped on top of a university dormitory in OK and although the numbers trapped with MBA were not significantly different compared to the control, they are reported in Table 3. The MBA lure had a significant impact on numbers of combined Fuscopolistes species trapped across all sites in WA, FL, GA, and SC, where replicate pairs were at least 30 m apart. For these sites, all wasps were more likely to be caught on MBA-baited traps (mean ± SEM: 6.73 ± 1.55) compared to unbaited control traps (1.95 ± 0.59) (χ 2 = 29.17, P < 0.001; Fig. 1). Table 3. MBA lure field test Wasp species and location . MBA . Control . Yakima Co.  P. aurifer 1.76 ± 0.28* 0.80 ± 0.20  P. aurifer (female) 0.57 ± 0.12* 0.26 ± 0.08 OK Towers  P. metricus 7.60 ± 2.67 3.30 ± 1.23  P. fuscatus 19.90 ± 5.81 14.70 ± 3.09  P. fuscatus (female) 7.60 ± 2.02 6.90 ± 1.32 FL, GA, SC Towers  P. metricus 15.82 ± 4.21* 2.91 ± 1.66  P. metricus (female) 12.00 ± 4.70* 3.81 ± 3.11  P. dorsalis 4.18 ± 1.27* 0.73 ± 0.49  P. bellicosus 5.27 ± 2.26* 0.36 ± 0.24 Wasp species and location . MBA . Control . Yakima Co.  P. aurifer 1.76 ± 0.28* 0.80 ± 0.20  P. aurifer (female) 0.57 ± 0.12* 0.26 ± 0.08 OK Towers  P. metricus 7.60 ± 2.67 3.30 ± 1.23  P. fuscatus 19.90 ± 5.81 14.70 ± 3.09  P. fuscatus (female) 7.60 ± 2.02 6.90 ± 1.32 FL, GA, SC Towers  P. metricus 15.82 ± 4.21* 2.91 ± 1.66  P. metricus (female) 12.00 ± 4.70* 3.81 ± 3.11  P. dorsalis 4.18 ± 1.27* 0.73 ± 0.49  P. bellicosus 5.27 ± 2.26* 0.36 ± 0.24 All wasps are males unless indicated otherwise. Mean ± SEM of wasps caught/trap during field tests in each location. * indicates a significant difference between numbers of wasps caught on traps with MBA lures compared to controls at P < 0.05. Open in new tab Table 3. MBA lure field test Wasp species and location . MBA . Control . Yakima Co.  P. aurifer 1.76 ± 0.28* 0.80 ± 0.20  P. aurifer (female) 0.57 ± 0.12* 0.26 ± 0.08 OK Towers  P. metricus 7.60 ± 2.67 3.30 ± 1.23  P. fuscatus 19.90 ± 5.81 14.70 ± 3.09  P. fuscatus (female) 7.60 ± 2.02 6.90 ± 1.32 FL, GA, SC Towers  P. metricus 15.82 ± 4.21* 2.91 ± 1.66  P. metricus (female) 12.00 ± 4.70* 3.81 ± 3.11  P. dorsalis 4.18 ± 1.27* 0.73 ± 0.49  P. bellicosus 5.27 ± 2.26* 0.36 ± 0.24 Wasp species and location . MBA . Control . Yakima Co.  P. aurifer 1.76 ± 0.28* 0.80 ± 0.20  P. aurifer (female) 0.57 ± 0.12* 0.26 ± 0.08 OK Towers  P. metricus 7.60 ± 2.67 3.30 ± 1.23  P. fuscatus 19.90 ± 5.81 14.70 ± 3.09  P. fuscatus (female) 7.60 ± 2.02 6.90 ± 1.32 FL, GA, SC Towers  P. metricus 15.82 ± 4.21* 2.91 ± 1.66  P. metricus (female) 12.00 ± 4.70* 3.81 ± 3.11  P. dorsalis 4.18 ± 1.27* 0.73 ± 0.49  P. bellicosus 5.27 ± 2.26* 0.36 ± 0.24 All wasps are males unless indicated otherwise. Mean ± SEM of wasps caught/trap during field tests in each location. * indicates a significant difference between numbers of wasps caught on traps with MBA lures compared to controls at P < 0.05. Open in new tab Fig. 1. Open in new tabDownload slide (A) An example replicate pair of traps from a FL air traffic control tower location. Species on the trap include P. metricus, P. dorsalis, and P. bellicosus. (B) Average Fuscopolistes trapped with MBA lures (6.73 ± 1.55) and control lures (1.95 ± 0.60) in WA, GA, SC, and FL (n = 81 replicate pairs). Values are significantly different (P < 0.001). Fig. 1. Open in new tabDownload slide (A) An example replicate pair of traps from a FL air traffic control tower location. Species on the trap include P. metricus, P. dorsalis, and P. bellicosus. (B) Average Fuscopolistes trapped with MBA lures (6.73 ± 1.55) and control lures (1.95 ± 0.60) in WA, GA, SC, and FL (n = 81 replicate pairs). Values are significantly different (P < 0.001). Discussion An objective of recent studies of paper wasp chemical ecology has been to identify and develop attractant semiochemicals to mitigate pest wasp populations. The venom volatiles of multiple Polistes species have been characterized (Bruschini 2006b, Weston et al. 1997), and N-(3-methylbutyl)acetamide (also reported as N-isopentylacetamide) has now been found to be a dominant volatile in the venom of nine species of Polistes and is present in both workers and gynes. We selected MBA as a hypothetical attractant to test in the field because of its consistent presence reported herein in the venom of all of the principal species of Polistes found in swarms and aggregations at towers in the southeastern United States (Reed and Landolt 1991), because of its presence in the venom of other Polistes species, and the attractiveness of venom glands to males in laboratory assays (Post and Jeanne 1983, 1984). The attractive nature of MBA to male paper wasps in the field was demonstrated for P. aurifer, P. metricus, P. bellicosus, and P. dorsalis. Significant attraction of male P. metricus and other Fuscopolistes males to this compound in field tests is consistent with results of laboratory studies indicating the venom as the source of an attractant and copulatory stimulant for males (Post and Jeanne 1983, MacKenzie et al. 2008, Reed and Landolt 1990). The attraction of female P. aurifer and P. metricus to MBA in the field was unanticipated, but could play a role in the formation and maintenance of heterospecific female aggregations observed on towers (Reed and Landolt 1991) (although it cannot be ruled out that the females were responding to other signals from males captured on traps). Female Polistes typically overwinter solitarily or in small, conspecific clumps (West-Eberhard 1969), and the large heterospecific aggregations and overwintering communities at towers in the southeastern United States where males are also active is exceptional (Reed and Landolt 1991). Chemically mediated gregarious behavior that assembles sexes for mating is well documented in the Insecta (Ali and Morgan 1990). Moreover, Polistes are known to mark regularly used overwintering sites with venom (Turillazzi et al. 2006). Venom volatiles, including MBA as demonstrated here, may mediate aggregation behavior of Polistes fall gynes by recruiting females to suitable overwintering sites once discovered. Nonetheless, the attraction of multiple Polistes species and sexes to MBA in the field aids in understanding the observations of heterospecific paper wasp aggregations present at towers and other conspicuous landscape features. Further testing of MBA as a trap lure is needed to better determine the potential and extent of its effectiveness for different Polistes species, sexes, and castes. Optimization of other operational parameters is needed to determine the best release rate for MBA lures with effective field lifetimes, and a superior dispenser for trap lures. Trap catch ratios in WA and southeastern/OK field tests may have varied due to the density of wasps at trapping sites (i.e., dense tower swarms vs dispersed males on patrol routes). Better information is also needed regarding optimum trapping sites and trapping times during the reproductive season as these contextual factors have important impacts on attraction. Trap design is another important element that needs further investigation. The adhesive traps we used in the field were developed and are sold as wasp traps, which could have affected wasp visual responses to the traps, perhaps decreasing differences between treatments and controls. Furthermore, Polistes species may possess multicomponent sex pheromones (Ayasse 2001), and field testing of other venom compounds in conjunction with MBA may lead to stronger and more species-selective lures. Despite these further research needs, our results indicate a clear potential for MBA as a lure in traps for the practical use of reducing numbers of these wasps when and where they are pestiferous at towers and tall buildings during autumn (Reed and Landolt 1991). The presence of MBA in venom is much wider taxonomically within the Polistes genus, and it is also found in other social wasp genera within the family Vespidae, suggesting it may be involved in the chemical ecology of a wide range of social wasp species (Aldiss 1983, Heath and Landolt 1988, Bruschini et al. 2010, Jimenez et al. 2016). Volatiles of the venom of Polistes species have been shown to be involved in alarm communication (Turillazzi 2006) in P. dominula and P. gallicus (Bruschini et al. 2006a, Bruschini et al. 2008a), and P. canadensis (Jeanne 1982). Thus, functions of volatile components of the venom of female Polistes, including MBA, comprise at least attractant and alarm functions. The same chemical(s) might be involved in additional functions depending on context, caste, blend components, and concentration (Bruschini et al. 2008b). Accordingly, our study revealed differences in amount of MBA in the sting apparatus between workers and fall gynes in P. dorsalis. Quantitative and qualitative differences in venom components between castes suggest that venom volatiles like MBA may serve multiple roles regulating different aspects of paper wasp behavior. The complex and diverse mating behavior of Polistes is prime for the involvement of chemical signals. As shown here, MBA is a strong candidate for a chemical compound that mediates the aggregation of Fuscopolistes gynes and males for mating. Evidence from our field tests suggest that MBA is a promising attractant for male and female P. aurifer, male and female P. metricus, and males of other Fuscopolistes that are common pests on man-made towers. Further work should seek to clarify the relative communicative value of other compounds in the venom that elicit attraction. This should lend insight into reproductive isolation and species recognition factors. Identification and field testing of additional venom compounds across Polistes may reveal synergists which enhance wasp responses to MBA; improving attractiveness and species selectivity, and augmenting trapping efforts to mitigate paper wasps under pestiferous circumstances. Acknowledgments We thank J. Brumley and A. Kenny-Chapman for technical assistance. We thank Sterling International Inc. of Spokane Valley, WA for contributing traps for field trials. Thanks to Dr. Rob Meagher at the USDA-ARS CMAVE in Gainesville, FL for providing physical facilities and help with the collection of wasps. We thank Megan Asche for collecting wasps in Florida and for support with dissections. We appreciate the efforts of Jillian Blake at Oral Roberts University for her help in running the Tulsa, OK experiments. Thanks to Dr. John Adamczyk and Dr. Chris Werle at the USDA-ARS Southern Horticultural Research Station for sharing lab space and aiding in wasp collection and logistical support. Thanks to Jerry Warner of the Central Washington Agricultural Museum for letting us collect wasps on museum property. We appreciate Robert A. Crowe of the Mississippi Army National Guard for letting us collect wasps at Camp Shelby. We greatly appreciate the thoughtful and constructive comments from Dr. J.G. Millar and Dr. Q.H. 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This work is written by (a) US Government employee(s) and is in the public domain in the US. Published by Oxford University Press on behalf of Entomological Society of America 2020.
TI - The Venom Compound N-(3-methylbutyl)acetamide Attracts Several Polistes (Fuscopolistes) Species (Hymenoptera: Vespidae)
JF - Journal of Economic Entomology
DO - 10.1093/jee/toaa065
DA - 2020-06-06
UR - https://www.deepdyve.com/lp/oxford-university-press/the-venom-compound-n-3-methylbutyl-acetamide-attracts-several-polistes-IwZgrFHC1f
SP - 1073
EP - 1079
VL - 113
IS - 3
DP - DeepDyve
ER -