62, P < 0.001), but no interaction between treatments and
colonies was verified (F4, 81 = 0.82, P = 0.52); the three colonies exhibited the same pattern of encapsulation rate variation ( Fig. 1). The encapsulation rates of workers whose actinobacteria were removed by streptomycin or a combination of streptomycin + penicillin were reduced in comparison with control workers, brush-treated or penicillin-treated workers (Fig. 2). Ten days after treatment, we could verify that the treatment had a highly significant effect (F5, 72 = 8.92, P < 0.001). We compared the survival proportion of the ants undergoing the bacteria removal treatments against that observed in the control groups. The hypothesis tested was H0: P control = P treatment
vs. H1: P control > P treatment (one-sided test). The TGF-beta Smad signaling p-value is computed based on the t-value for the following comparisons: buy VX-809 Control vs. Dry brush, P = 0.0042; Control vs. Wet brush, P = 0.0001; Control vs. Pen. G, P = 0.0021; Control vs. Strep., P = 0.0021; Control vs. Pen. G + Strep., P = 0.0002. As all treatments provoked mortality in treated ants, including the Dry brush, it appears that ant mortality is due to the stress of the ant removal from the nest and its manipulation. It is possible that the treatments to eliminate actinobacteria cause selective survival; therefore, we would be sampling the encapsulation response of a subset of the ants. However, we have no evidence of differential mortality associated
with the level of encapsulation response because similar mortality occurred in groups with higher encapsulation response (Wet brush) and Vildagliptin in groups with lower encapsulation response (Pen. G + Strep.), as verified in Fig. 2. The individual metabolic rate of the workers, measured in terms of CO2 production, showed a pattern of increase as workers lost their bacterial coating and switched to external activities (Fig. 3; Kruskal–Wallis, H (2 n = 42) = 6.94, P = 0.03). Individuals living inside the nest, with or without a whitish coat of bacteria, had significantly lower respiration rates compared with individuals performing external activities. Hydrocarbon quantities on the thorax did not vary among the three groups: 119.8 ± 27.7 ng per ant (mean ± SE) for EXT, 81.1 ± 11.0 for INØ and 132.3 ± 32.8 for INB (Kruskal–Wallis H (2, n = 53) = 1.67, P = 0.43) (See Fig. S1). The hydrocarbon profile was simple (24 peaks, see Fig. S2). The hydrocarbons observed were mainly methyls (11- + 13- + 15-MeC29, more than 30%, see Table S1; 11- + 13-MeC31–10%) and the corresponding dimethyls (respectively 11,15- + 13,17-DiMeC29, 5% and 11,15- + 13,17-DiMeC31, 6%), and the hydrocarbon profile was not changed according to the ant group. In the dendrogram, the samples were mixed in arbitrary groups (see Fig. S3). We found some of the ant hydrocarbons in the bacteria and also in the gelose (see Table S1), but in very small quantities (4.5 and 9.7 ng, respectively).