Parasitic wasps have long been thought to be unable to synthesize fatty acids de novo, but recent 13 C-labelling studies have challenged this view. It remained unclear, however, whether the reported biosynthesis rates are of biological relevance. Here, we show in Nasonia vitripennis that ageing females with partly depleted lipid reserves produce biologically relevant amounts of fatty acids de novo. Females with varying oviposition history (0–48 h) prior to feeding 20% 13 C-labelled glucose solution showed 13 C-incorporation rates of (mean ± SEM) 30 ± 2%, 50 ± 2%, 49 ± 3% and 21 ± 2% in palmitic, stearic, oleic and linoleic acid, respectively. The absolute amounts of fatty acids synthesized de novo across treatments corresponded to 28 ± 3 egg lipid equivalents. Females incorporated de novo synthesized fatty acids into their eggs, and glucose-fed females laid more eggs than water-fed control females. The number of eggs laid prior to glucose feeding did not correlate with the degree of lipogenesis, but the amounts of de novo synthesized fatty acids correlated with constitutive (not synthesized de novo) fatty acids. Hence, glucose feeding has a twofold effect on the fatty acid status of N. vitripennis females by decelerating the catabolism of existing fat reserves and partially replenishing ebbing fat reserves by lipogenesis.