Online Resource 3Identification of the putative aggregation pheromone components emitted by the European earwig, Forficula auricularia

Stephen R. Quarrell1· Noel W. Davies2 · Paul W. Walker1· Stephen R. Corkrey1· Jason A. Smith3· Geoff R. Allen1

1School of Land and Food/ Tasmanian Institute of Agriculture, University of Tasmania, Private Bag 98, Hobart 7001, Australia e-mail: 2Central Science Laboratory, University of Tasmania, Private Bag 74, Hobart 7001, Australia; 3School of Physical Sciences, University of Tasmania, Private Bag 75, Hobart 7001, Australia

Online Resource 3aMean (±SEM) male earwigs per trap per tree and mean (±SEM) treatment effect (treatment – hexane control) to hydrocarbons after a 12 hour period in field based experiments. Positive numbers indicate attraction. Negative numbers indicate repellency. Compounds were tested within apple and cherry trees (n = 20) in a paired design against hexane controls. Bold type indicates significant difference Wilcoxon sign rank < 0.05.

Field experiment date
Compound / Concentration / 6th Jan 2012 / 19th Jan / 28th Jan / 3rd Feb / 23rd Feb / 2nd Dec / 7th Dec / 22nd Dec / 6th Feb 2013
Mean males/trap / 0.34 (0.07) / 0.23 (0.06) / 0.67 (0.11) / 2.60 (0.33) / 1.13 (0.14) / 0.00 (0.00) / 0.74 (0.13)
Male wash / 10 IE# / -0.25 (0.23) / 0.05 (0.15)
Female wash / 10 IE# / -0.15 (0.21)
n-alkane blenda / 0.1mg / 0.40 (0.15)
HC blend 2b / 0.1mg / 0.00 (0.18)
(Z)-9-C23 / 0.1mg / 0.30 (0.26)
(Z)-7-C23 / 0.1mg / -0.10 (0.22)
(Z)-9-C25 / 0.1mg / 0.10 (0.29)
(Z)-7-C25 / 0.1mg / -0.20 (0.32)
alkene blend 3c / 0.05mg / -0.15 (0.38) / 0.00 (0.00) / -0.05 (0.20) / -0.05 (0.05) / -0.05 (0.20)
alkene blend 3c / 0.1mg / 0.15 (0.17) / 0.3 (0.21) / 0.35 (0.38) / -0.05 (0.48) / 0.25 (0.43) / 0.00 (0.00) / 0.25 (0.44)
alkene blend 3c / 0.2mg / 0.1 (0.12) / -0.30 (0.44) / -0.50 (0.39) / -0.01 (0.07) / -0.50 (0.39)
alkene blend 4d / 0.05mg / 0.35 (0.41) / 0.00 (0.00)
alkene blend 4d / 0.1mg / 1.55 (0.85) / 0.15 (0.32) / -0.50 (0.22) / 0.05 (0.05) / -0.5 (0.22)
alkene blend 4d / 0.2mg / -0.15 (0.35)
alkene blend 5e / 0.1mg / 0.65 (0.37)
alkene blend 5e / 0.2mg / 0.20 (0.28)

IE = Insect Equivalents, * 1 hour hexane extraction, ^ 3 x 100 µL cuticular hexane wash

an-alkane blend; n-C21 : n-C23 : n-C25; Blendratio: 40:85:70

b Seven component blend n- C21 ; (Z)-9-C23 : (Z)-7-C23 : n- C23 : (Z)-9-C25 : (Z)-7-C25 : n- C25; Blend ratio: 40: 70:20:85:80:15:70

c Four component blend (Z)-9-C23 : (Z)-7-C23 : (Z)-9-C25 : (Z)-7-C25; Blend ratio: 60:15:100:25

d Two component blend (Z)-9-C23 : (Z)-9-C25;Blendratio: 30:70

e Two component blend (Z)-7-C23 : (Z)-7-C25;Blendratio: 30:70

Online Resource 3bMean (±SEM) female earwigs per trap per tree and mean (±SEM) treatment effect (treatment – hexane control) to hydrocarbons after a 12 hour period in field based experiments. Positive numbers indicate attraction. Negative numbers indicate repellency. Compounds were tested within apple and cherry trees (n = 20) in a paired design against hexane controls. Bold type indicates significant difference Wilcoxon sign rank < 0.05.

Field experiment date
Compound / Concentration / 6th Jan 2012 / 19th Jan / 28th Jan / 3rd Feb / 23rd Feb / 2nd Dec / 7th Dec / 22nd Dec / 6th Feb 2013
Mean females/trap / 0.53 (0.09) / 0.36 (0.07) / 1.08 (0.16) / 0.53 (0.12) / 1.65 (0.19) / 0.03 (0.03) / 1.13 (0.18)
Male wash / 10 IE# / -0.35 (0.20) / 0.10 (0.19)
Female wash / 10 IE# / -0.20 (0.17)
n-alkane blenda / 0.1mg / -0.20 (0.16)
HC blend 2b / 0.1mg / 0.35 (0.32)
(Z)-9-C23 / 0.1mg / 0.00 (0.39)
(Z)-7-C23 / 0.1mg / -0.09 (0.55)
(Z)-9-C25 / 0.1mg / 0.35 (0.53)
(Z)-7-C25 / 0.1mg / -0.50 (0.48)
alkene blend 3c / 0.05mg / 0.60 (0.38) / 0.00 (0.00) / 0.15 (0.23) / 0.00 (0.00) / 0.15 (0.23)
alkene blend 3c / 0.1mg / 0.50 (0.30) / -0.10 (0.24) / 0.90 (0.52) / 0.55 (0.78) / -0.65 (0.44) / 0.05 (0.05) / -0.65 (0.43)
alkene blend 3c / 0.2mg / 0.05 (0.17) / 0.25 (1.12) / -0.15 (0.64) / 0.15 (0.08) / -0.15 (0.64)
alkene blend 4d / 0.05mg / 0.70 (0.44) / 0.01 (0.07)
alkene blend 4d / 0.1mg / 0.55 (0.60) / -0.15 (0.50) / 0.15 (0.45) / 0.05 (0.05) / 0.15 (0.43)
alkene blend 4d / 0.2mg / 0.55 (0.65)
alkene blend 5e / 0.1mg / 1.05 (0.41)
alkene blend 5e / 0.2mg / 0.00 (0.29)

#IE = Insect Equivalents, * 1 hour hexane extraction, ^ 3 x 100 µL cuticular hexane wash

an-alkane blend; n-C21 : n-C23 : n-C25; Blendratio: 40:85:70

b Seven component blend n- C21 ; (Z)-9-C23 : (Z)-7-C23 : n- C23 : (Z)-9-C25 : (Z)-7-C25 : n- C25; Blend ratio: 40: 70:20:85:80:15:70

c Four component blend (Z)-9-C23 : (Z)-7-C23 : (Z)-9-C25 : (Z)-7-C25; Blend ratio: 60:15:100:25

d Two component blend (Z)-9-C23 : (Z)-9-C25;Blendratio: 30:70

e Two component blend (Z)-7-C23 : (Z)-7-C25;Blendratio: 30:70

Online Resource 3cMean (±SEM) juvenile earwigs (2nd, 3rd and 4th instars) per trap per tree and mean (±SEM) treatment effect (treatment – hexane control) to hydrocarbons after a 12 hour period in field based experiments. Positive numbers indicate attraction. Negative numbers indicate repellency. Compounds were tested within apple and cherry trees (n = 20) in a paired design against hexane controls. Bold type indicates significant difference Wilcoxon sign rank < 0.05.

Field experiment date
Compound / Concentration / 6th Jan 2012 / 19th Jan / 28th Jan / 3rd Feb / 23rd Feb / 2nd Dec / 7th Dec / 22nd Dec / 6th Feb 2013
Mean juveniles/trap / 0.52 (0.10) / 0.35 (0.09) / 0.76 (0.14) / 2.53 (0.57) / 0.04 (0.02) / 8.91 (1.48) / 0.49 (0.10)
Male wash / 10 IE# / 0.23 (0.33) / 0.25 (0.33)
Female wash / 10 IE# / -0.10 (0.19)
n-alkane blenda / 0.1mg / -0.03 (0.36)
HC blend 2b / 0.1mg / -0.05 (0.42)
(Z)-9-C23 / 0.1mg / -2.40 (0.12)
(Z)-7-C23 / 0.1mg / -3.20 (0.30)
(Z)-9-C25 / 0.1mg / -2.90 (0.15)
(Z)-7-C25 / 0.1mg / -3.80 (0.20)
alkene blend 3c / 0.05mg / 0.15 (0.16) / 5.63 (3.98) / -0.23 (0.28) / -1.18 (2.76) / -0.23 (0.28)
alkene blend 3c / 0.1mg / 0.30 (0.41) / 0.20 (0.30) / -3.55 (0.18) / 0.23 (0.26) / 0.18 (0.26) / -1.93 (3.34) / 0.18 (0.26)
alkene blend 3c / 0.2mg / 0.48 (0.33) / -2.45 (0.12) / -0.03 (0.45) / 0.90 (3.65) / -0.03 (0.45)
alkene blend 4d / 0.05mg / 0.03 (0.05) / 1.38 (1.73)
alkene blend 4d / 0.1mg / -0.08 (0.27) / -0.03 (0.14) / 0.15 (0.29) / 1.40 (3.45) / 0.15 (0.29)
alkene blend 4d / 0.2mg / 0.00 (0.00)
alkene blend 5e / 0.1mg / 0.00 (0.10)
alkene blend 5e / 0.2mg / 0.05 (0.10)

#IE = Insect Equivalents, * 1 hour hexane extraction, ^ 3 x 100 µL cuticular hexane wash

an-alkane blend; n-C21 : n-C23 : n-C25; Blendratio: 40:85:70

b Seven component blend n- C21 ; (Z)-9-C23 : (Z)-7-C23 : n- C23 : (Z)-9-C25 : (Z)-7-C25 : n- C25; Blend ratio: 40: 70:20:85:80:15:70

c Four component blend (Z)-9-C23 : (Z)-7-C23 : (Z)-9-C25 : (Z)-7-C25; Blend ratio: 60:15:100:25

d Two component blend (Z)-9-C23 : (Z)-9-C25;Blendratio: 30:70

e Two component blend (Z)-7-C23 : (Z)-7-C25;Blendratio: 30:70