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Trivers R. Turrillazi S. Venkataraman A. Visscher P. Wcislo W. West-Eberhard M. Wheeler D. Wheeler W. Wilson E. Yamane S. Personalised recommendations. A total of 50 assays were performed across the nine species Table 1. Following Smith et al. Two-sample t-tests were performed between pairs of social complexity and nest type categories.
To standardise measurements across different species, concentrations of wasp equivalents were converted to concentrations of equivalent surface area. Adapting methods previously applied to bees  , mean surface area for each species was estimated by generating elliptical cylinders using measurements from up to ten individuals per species. Amplification of the 28S gene fragments was performed using primers previously used in the construction of microgastrid wasp phylogenies  and COI gene fragments amplifications were performed using generic invertebrate primers for that region .
Following PCR amplicons were purified using ExoSap-IT USB according to the manufacturer instructions and purified products were sequenced using their corresponding forward primers with dye terminator reactions on a x1 Genetic Analyser Applied Biosystems. Phylogenetic reconstruction of nine wasp species plus Apis mellifera as an outgroup was performed using a bp sequence generated by concatenating the two gene fragments: 28S nrDNA bp and COI mtDNA bp.
Generated sequences Table 2 plus corresponding sequences from A. Length polymorphisms in sequence data were removed following alignment and prior to concatenation of gene fragments.
Phylogenies were created using both distance-based neighbour-joining and Bayesian methods maximum clade credibility. Neighbour-joining phylogenetic reconstruction was performed in MEGA v4. Both transition and transversion substitutions were included, assuming homogeneous patterns among lineages and uniform rates among sites. Gaps were treated as complete deletions. Bootstrap values were obtained using replicates.
Trees were generated using a single MCMC chain of 20 million steps sampling every steps. The molecular clock rate was fixed to 1. All other parameters were left in their default state as generated by BEAUti.
Integrity of generated data was checked using Tracer v1. TreeAnnotator v1. IC50 values could be calculated for 44 of the 50 assays performed Table 1. Two of the five assays performed using extract obtained from Austroscolia sp. Similarly these species were not included when calculating differences in mean IC50 by social complexity.