MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13712/m_MTBLS13712_GC-MS_positive__metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13712/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13712/a_MTBLS13712_GC-MS_positive__metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13712/s_MTBLS13712.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13712 CHC compounds were identified by the compound-specific retention indices and their detected diagnostic ions (Carlson et al. 1998). MetaboLightsPublicGas Chromatography MS - positiveWe analysed the CHC extracts of all samples with an Agilent 6890 gas chromatograph coupled with an Agilent 5975 Mass Selective Detector (GC-MS, Agilent, Waldbronn, Germany):  The GC (split/splitless injector in splitless mode for 1 min, injected volume 1 μl at 300°C) was equipped with a DB-5 Fused Silica capillary column (30 m x 0.25 mm ID, df = 0.25 µm; J&W Scientific, Folsom, USA). Helium served as carrier gas at a constant flow of F. The following temperature program was usedStart temperature 60 °C, temperature increase by 5 °C per min up to 300 °C, isotherm at 300 °C for 10 min.Coming out from the shadows: facultative slave-making ants reveal their chemical identity during colony development. 10.1007/s00265-026-03738-0.Tomasz WlodarczykUniversity of BialystokCuticlepure substancemass spectrometry assayAfter removal, ants were killed by freezing and stored at −22 ºC for subsequent chemical analyses. A total of 78 mature F. fusca ants, 65 mature and 32 callow F. sanguinea ants were used for CHC profiling. The F. sanguinea colonies also served as a source of pupae for the separation experiments in which pupae were separated from the parent colonies and reared in isolation. The separated individuals were sacrificed for CHC analysis at different time intervals after eclosion (1-3 days, 8-10 days, 17-20 days and 35-40 days). In the “dummy ant” experiment, separated ants were kept with glass beads coated with CHCs extracted from either F. sanguinea or F. fusca workers. Moreover, workers from 12 out of 19 free-living F. fusca colonies used as sources for slaves were also sampled for chemical analyses (five to eight ants per colony). Individual ants were placed in glass vials (2 ml) and extracted in 150 μl of hexane for 10 minutes. Subsequently, 15 μl of a hexane solution of docosane (12.5 μg/ml) was added to the extracts as an internal standard. Glass vials with the extracts were left open until the solvent evaporated, re-dissolved in 50 μl of hexane, transferred to 100 μl inserts, and stored at -22 ºC until analysis. reference compoundFormica fuscaFormica sanguineahttps://www.ebi.ac.uk/metabolights/MTBLS13712Thomas Schmitt.Tomasz Włodarczyk. University of Białystok. ul.Swierkowa 20B, 15-328 Bialystok. tomwlo@gmail.com.Chromatograms and mass spectra were recorded and quantified via integrated peak areas with the software HP Enhanced ChemStation G1701AA (version A.03.00; Hewlett Packard).Sanguinea proportionTreatmentExperimentAge statusColony IDSampling datetomwlo@gmail.comAnts were collected from laboratory colonies (colony development experiment). Part of the ants were separated on Petri dishes prior to collection (separation and dummy ant experiment). Sixteen ant colonies were established from F. sanguinea queens provided with 80 to 230 F. fusca pupae. Colonies were maintained in the plastic boxes (40 × 30 × 30 cm) with the floor covered with a thin layer of mineral soil and sawdust. Test tubes wrapped in aluminium foil and partially filled with water, and closed with a cotton plug, served nesting sites for ants. The inner surface of the nest box walls was coated with Fluon to prevent ants from escaping. Colonies were fed diluted honey, fresh apple pieces, as well as crickets (Acheta domesticus), greater wax moth (Galleria mellonella) caterpillars, and male honey bee larvae and pupae all killed by freezing. Each of the developing F. sanguinea colonies was subjected to at least one census followed by the removal of 3-6 slave-maker workers (if present) and a similar number of F. fusca slaves. The timing of censuses was chosen to cover various stages of colony development spanning a range of slave-to-slave-maker ratios. Some of the samples included callow slave-maker workers, which were identified by the brighter coloration due to the incomplete cuticle pigmentation. MetabolomicsMetabolomicsgas chromatography-mass spectrometryuntargeted analysisInsectaAgilent 5975 MSDchemical ecologyCuticleAgilent 6890 GCFormica fuscaFormica sanguineapure substancereference compound mixexperimental samplereference compoundcuticular hydrocarbonsMetabolomicsgas chromatography-mass spectrometryuntargeted analysisInsectaAgilent 5975 MSDchemical ecologyCuticleAgilent 6890 GCFormica fuscaFormica sanguineapure substancereference compound mixexperimental samplereference compoundcuticular hydrocarbonsThe electron ionisation mass spectra (EI-MS) were acquired at an ionisation voltage of 70 eV (source temperature: 230°C). n-pentacosanen-docosanefalseCuticular hydrocarbon profiles of Formica sanguinea and and their Formica fusca slaves over the colony developmentOne of the main challenges of the socially parasitic mode of life is bypassing the host's recognition ability, which ensures that altruistic behaviour is directed towards related individuals. Various chemical strategies have evolved to achieve this goal. The most widespread, used also by the obligate slave-making ants, is camouflage or mimicry of colony odour encoded in cuticular hydrocarbon (CHC) composition. However, recent studies have shown that facultative slave-makers employ a different strategy: they manipulate the slaves' recognition labels to make them resemble the parasite's CHC profile. We examined the limitations of this strategy by focusing on incipient F. sanguinea colonies, where slaves are the majority. Our study revealed that callow F. sanguinea ants initially suppress their species-specific odour profile, which develops gradually over time accompanied by an increase of CHC amount per surface area in slave-maker workers. This allows the slaves to familiarise themselves with the parasite's CHC. We found that callow ants produce lower amounts of CHC, and the relative abundance of certain compounds differs from what is observed in older ants. Additionally, preimaginal stages of F. sanguinea ants acquire CHC from the slaves, which are later incorporated into the imagines’ recognition labels. These findings support the proposition that the parasite's manipulation strategy is limited by the slaves' learning capacity, which is necessary to maintain colony cohesion. They also shed light on the selective pressures that might have led to the evolution of chemical mimicry in mature obligate slave-maker colonies.2026-05-192026-01-17MTBLS13712MTBLC32938MTBLC46050CHEBI:32938CHEBI:46050