DOI: 10.5061/dryad.hqbzkh1j6
title: Data from: Targeted treatment of injured nestmates with antimicrobial compounds in an ant society

description: Infected wounds pose a major mortality risk in animals. Injuries are
 common in the ant Megaponera analis, which raids pugnacious prey. Here we
 show that M. analis can determine when wounds are infected and treat them
 accordingly. By applying a variety of antimicrobial compounds and proteins
 secreted from the metapleural gland to infected wounds, workers reduce the
 mortality of infected individuals by 90%. Chemical analyses showed that
 wound infection is associated with specific changes in the cuticular
 hydrocarbon profile, thereby likely allowing nestmates to diagnose the
 infection state of injured individuals and apply the appropriate
 antimicrobial treatment. This study demonstrates that M. analis ant
 societies use antimicrobial compounds produced in the metapleural glands
 to treat infected wounds and reduce nestmate mortality.
description: Chemical Ecology: Cuticular Hydrocarbon
 analysis (GC-MS data) Metapleuralgland secretions
 (TD-GC-MS data)
description: # Targeted treatment of injured nestmates with antimicrobial compounds in
 an ant society Data
 includes the 1) finished data table for the Cuticular hydrocarbon (CHC)
 analyses; 2) the raw data created by the GC-MS for the CHC analyses and 3)
 the raw data created from the TD-GC-MS for the Metapleural gland
 secretions (6890 gas chromatograph (GC) coupled to a 5975 mass selective
 detector (MS) by Agilent Technologies (Waldbronn, Germany). The folders
 for each data point contain all the information which is readable by the
 Softwares Chemstation or Masshunter Qualitative Analysis (Agilent
 Technologies). ## Description of the data and file structure The finished
 data table (1) contains the cleaned up datatable of the raw CHC data (2).
 In the datatable we identified the Hydrocarbon Compound (Compound), the
 type of compound (type), the Retention time the compound appears on the
 Gas Chromatograph in minutes (Ret Time) and the standardised Retention
 Index (Ret Index). The Cuticular Hydrocarbon (CHC) Sample IDs are composed
 of my initials (EF: Erik Frank), the year (20: 2020) and their individual
 ID (1 to 60). The numbers received different treatments: 1-6 CHC profile
 extracted at timepoint 0 of infected ants. 7-18 CHC profile extracted
 after 2hours of infected ants. 19-30 CHC profile extracted after 11 hours
 of infected ants. 31-36: CHC profile extracted at timepoint 0 of sterile
 ants. 37-48 CHC profile extracted after 2hours of sterile ants. 49-61 CHC
 profile extracted after 11 hours of sterile ants. Infected ants refer to
 ants whose leg injury was exposed to Pseudomonas aeruginosa, while for
 sterile ants the leg injury was exposed to a sterile PBS solution. In
 addition we ran conditioning blanks to assure the purity of the GC-MS
 machine and the quality of the hexane and remove any potential
 contaminants. These are in the sub-folder "conditioning blanks"
 and named Hexane_1 to 6. Between the runs we also ran additional Hexane
 samples to assure the quality was maintained throughout the run. These are
 named "Hexane_1" and Hexane_Blank_2 to 4. The Metapleuralgland
 samples are named after the species *Megaponera analis* from which they
 were taken from (M_analis), followed by the gland abbreviation (mpg) and
 sample number (1 to 3). Each sample contains 6 pooled metapleural glands
 taken from 6 individual worker ants. The sample "test after mpg"
 was a TD-GC-MS run containing an empty glass tube to identify contaminants
 or potential remnants from the sample runs. To quantify differences in CHC
 profiles between infected and sterile ant workers, we placed the ants in
 hexane for 10 minutes to extract the CHC profile. The cuticular
 hydrocarbon extracts were then evaporated to a volume of approximately 100
 μL and 1 μL was analyzed by using a 6890 gas chromatograph (GC) coupled to
 a 5975 mass selective detector (MS) by Agilent Technologies (Waldbronn,
 Germany). The GC was equipped with a DB-5 capillary column (0.25 mm ID ×
 30 m; film thickness 0.25 μm, J & W Scientific, Folsom, Ca, USA).
 Helium was used as a carrier gas with a constant flow of 1 mL/min. A
 temperature program from 60 °C to 300 °C with 5 °C/min and finally 10 min
 at 300 °C was employed. Mass spectra were recorded in the EI mode with an
 ionization voltage of 70 eV and a source temperature of 230 °C. The
 software ChemStation v. F.01.03.2357 (Agilent Technologies, Waldbronn,
 Germany) for windows was used for data acquisition. Identification of the
 components was accomplished by comparison of library data (NIST 17) with
 mass spectral data of commercially purchased standards and diagnostic
 ions. The Metapleuralgland samples were placed in a glass-wool-packed
 thermodesorption tube and placed in the thermodesorber unit (TDU;
 TD100-xr, Markes, Offenbach am Main, Germany). The thermodesorption tube
 was heated up to 260°C for 10 min. The desorbed components were
 transferred to the cold trap (5 °C) to focus the analytes using N2 flow in
 splitless mode. The cold trap was rapidly heated up to 310 °C at a rate of
 60 °C per minute, held for 5 min and connected to the GC-MS (Agilent 7890B
 GC and 5977 MS, Agilent Technologies, Palo Alto, USA) via a heated
 transfer line (300 °C). The GC was equipped with an HP-5MS UI capillary
 column (0.25 mm ID × 30 m; film thickness 0.25 μm, J & W Scientific,
 Folsom, Ca, USA). Helium was the carrier gas using 1.2874 ml/min flow. The
 initial GC oven temperature was 40 °C for 1 min, then raised at a rate of
 5°C per min until reaching 300 °C, where it was held for 3 minutes. The
 transfer line temperature between GC and MS was 300 °C. The mass
 spectrometer was operated in electron impact (EI) ionization mode,
 scanning m/z from 40 to 650, at 2.4 scans per second. Chemical compounds
 were identified using the same protocol as for the CHCs. ## Code/Software
 All Data was quantified and prepared using MassHunter Qualitative Analysis
 (Agilent Technologies) and Excel for Mac (Version 16.79.1).