Gut samples from Hi0 and Hi50 groups were collected at t0 and t1. Specifically, 9 different sturgeons per dietary group (n = 3) were collected and processed as previously described by Zarantoniello et al.38. The obtained cell pellets were covered with RNA later Stabilization Solution (Ambion, Foster City, CA, USA) and stored at − 80 °C until the extraction of total microbial RNA performed by Quick-RNA Miniprep kit (Zymo Research, CA, USA). The quantity and purity of the extracted RNA were checked using a Nanodrop ND 1000 (Thermo Fisher Scientific). Moreover, the absence of residual DNA contamination was checked by PCR as described by Garofalo et al.69. Each sample RNA (10 μL) was reverse-transcribed in cDNA using oligo (dT) and random hexamer primers from SensiFAST cDNA Synthesis Kit for RT-qPCR (Bioline, London, UK).

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