Supplementary material 6 from: Lapeva-Gjonova A, Gjonov I, Kostova R, Mifsud D (2025) Vibroacoustic signalling of the Maltese Messor ants (Hymenoptera, Formicidae), with geographical insights into M. ibericus. Journal of Hymenoptera Research 98: 1087-1106. https://doi.org/10.3897/jhr.98.170415

Supplementary material 6: Statistical analyses reports Table 1. Results of PERMANOVA test for di5erences in the acoustic characteristics between Messor species in Malta (significant di5erences are marked with asterisk - *). Source df SS MS Pseudo-F P(perm) Unique perms P(MC) Species 2 103.33 51.66 15.96 0.0001* 9949 0.0001* Res 33 106.81 3.24 Total 35 210.14 Table 2. Results of pairwise post-hoc test in PERMANOVA for di5erences in the acoustic characteristics between Messor species in Malta (significant di5erences are marked with asterisk - *). Groups t P(perm) Unique perms P(MC) Messor bouvieri, Messor capitatus 3.5179 0.0001* 8155 0.0001* Messor bouvieri, Messor ibericus 5.3423 0.0001* 9944 0.0001* Messor capitatus, Messor ibericus 3.0162 0.0006* 9947 0.0004* Table 3. Acoustic characteristics entered into the predictive discriminant model by SDA for Messor species in Maltese Islands (significant di5erences are marked with asterisk - *). Step Entered Wilks' Lambda Statistic df1 df2 df3 Exact F Statistic df1 df2 Sig. 1 Chirp duration (ms) 0.19 1 2 33.00 68.47 2 33 <0.001* 2 Amplitude forward/backward ratio 0.11 2 2 33.00 32.72 4 64 <0.001* 3 Backward chirp duraMon (ms) 0.07 3 2 33.00 28.32 6 62 <0.001 * 4 Chirp rate (chirp/s) 0.04 5 2 33.00 24.85 10 58 <0.001* Table 4. Cross-validated classification matrix for Messor species in Maltese Islands (number and percent classified individuals). Species Predicted Group Membership Total Messor bouvieri Messor capitatus Messor ibericus Count Messor bouvieri 9 0 0 9 Messor capitatus 0 9 0 9 Messor ibericus 0 0 18 18 % Messor bouvieri 100.00 0.00 0.00 100.0 Messor capitatus 0.00 100.00 0.00 100.0 Messor ibericus 0.00 0.00 100.00 100.0 Table 5. Results of PERMANOVA test for di5erences in the acoustic characteristics between Messor ibericus populations (significant di5erences are marked with asterisk - *). Source df SS MS Pseudo-F P(perm) Unique perms P(MC) Place 4 226.88 56.72 60.76 0.0001* 9935 0.0001* Res 40 37.34 0.93 Total 44 264.22 Table 6. Results of pairwise post-hoc test in PERMANOVA for di5erences in the acoustic characteristics between Messor ibericus populations (significant di5erences are marked with asterisk - *). Groups t P(perm) Unique perms P(MC) Sicily, Crete 2.5681 0.0025 8148 0.0028* Sicily, Malta 10.111 0.0003 8160 0.0001* Sicily, Gozo 8.8846 0.0001 8111 0.0001* Sicily, Bulgaria 11.204 0.0001 8168 0.0001* Crete, Malta 8.6473 0.0001 8208 0.0001* Crete, Gozo 8.5308 0.0001 8116 0.0001* Crete, Bulgaria 9.3738 0.0001 8224 0.0001* Malta, Gozo 6.4097 0.0001 8131 0.0001* Malta, Bulgaria 1.6688 0.057 8233 0.0633 Gozo, Bulgaria 6.3927 0.0001 8114 0.0001* Table 7. Acoustic caracteristics entered into the predictive discriminant model by SDA for the M. ibericus populations (significant di5erences are marked with asterisk - *). S t e p Entered Wilks' Lambda Statistic df 1 df 2 df 3 Exact F Approximate F Statistic df 1 df 2 Sig. Statistic df1 df2 Sig. 1 Chirp rate (chirp/s) 0.031 1 4 40 316.30 4 40 <0.001* 2 Backward freq. peak (Hz) 0.004 2 4 40 154.98 8 78 <0.001* 3 Chirp duration (ms) 0.002 3 4 40 84.00 12 100.83 <0.001* 4 Backward chirp duration (ms) 0.001 4 4 40 60.28 16 113.67 <0.001* 5 Amplitude forward/backw ard ratio 0.001 5 4 40 50.28 20 120.35 <0.001* 6 Forward chirp duraRon (ms) 0.000 6 4 40 41.63 24 123.31 <0.001* 7 Forward freq. peak (Hz) 0.000 7 4 40 36.24 28 124.01 <0.001*