Extended Data Fig. 5: No significant changes in maternal or offspring microbiome.

(a & b) Bray–curtis dissimilarities showing no change in the composition of the gut microbiome in (a) F1 offspring sired by nABX fathers or (b) in mothers mated with nABX males, indicating transmitted F1 effects are independent of influencing the F1 or maternal microbiota per se. (c) Microbiota beta diversity confirming no consistent alteration in F1 microbiota as a function of the fathers microbial status. (d) Rarefaction analysis in fathers, mothers and offspring stratified by treatment of the father. (e) Spearman analysis between offspring taxa abundance and offspring phenotype shows no correlation (left), whereas spearman analysis between paternal microbiota taxa abundance and offspring phenotype shows associations of specific taxa in fathers with positive or negative correlations to F1 phenotype. Grey dot indicates mean correlation. (f) Analysis of microbiota richness of mothers and fathers prior to - and following − 4d cohousing and mating with either a control or nABX male. Pre- and post-mating faecal samples were collected from both mating pairs (CON = 6; nABX = 6). The results indicate there is no significant change in mothers following 4 days exposure to nABX males. Bar represents median and whiskers indicate 1.5x IQR. (g) Foldchange in specific taxa in mothers during pre- to post- mating with either a control or nABX male, revealing high variance and no significant effects linked with nABX males. Faecal samples collected pre- and post-mating (CON = 6; nABX = 6, females). Error bars indicate 95% C.I. (h) Upper; qPCR analysis showing unaltered microbiota abundance in mothers post-mating and after 4d exposure to nABX males. Also shown is failure to detect a seminal fluid microbiome ruling out this modality of information transfer (CON = 8; nABX = 7, seminal fluid samples). Error indicates S.D. Lower: Bray–curtis dissimilarities in fathers and mothers from pre- and post- mating periods.