1. (A) Schematic of experimental timeline; details are as follows.
(A) Schematic of experimental timeline; details are as follows. Sow treatment groups comprised control (CON) (n = 9) and fecal microbiota transplantation procedure (FMTP) (n = 9) groups; the FMTP is described in Materials and Methods. “Pre-Ab” is preantibiotic, and “Post-Ab” is 4 days postantibiotic. On days 70 and 100, 200-ml fecal inocula were given to sows via gastric intubation. Post-first FMT fecal sampling was done 30 days after the first FMT, i.e., on day 100 of gestation. Post-second FMT fecal sampling was done 12 days after the second FMT, i.e., on day 112 of gestation. FMT1 treatment comprised a single 8-ml fecal inoculum given orally to offspring at birth, and FMT4 treatment comprised 8-ml fecal inocula given orally to offspring at birth and at 3, 7, and 28 days of age. (B) Schematic of fecal inoculum preparation; details are as follows. Pooled fecal samples were homogenized in maximum recovery diluent (MRD; Oxoid Ltd.) by stirring at 1,000 rpm for 10 min using a magnetic stirrer and then strained through a sieve to remove large particles, which were then washed through the sieve with another part of MRD. The homogenate was centrifuged for 30 min at 4,000 × g, and the supernatant discarded. Aliquots of the pellets in MRD + glycerol were stored at 5°C to allow microbial adjustment to low temperatures and then stored for no longer than 3 months at −80°C. (C) Schematic of study design and sample analysis.
Ursula M. McCormack et al. mSystems 2018; doi: /mSystems