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Determination of microbiome Microbial DNA was extracted from fecal samples using stool transport and recovery (STAR) buffer (Roche, Basel, Switzerland) and SpheroLyse solution (Hain Lifescience, Nehren, Germany)

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The cell pellet was thawed, homogenized, and lysed in a buffer containing 20 mM HEPES pH 7.5, 50 mM NaCl, 2 mM MgCl 2 , and complete protease inhibitor cocktail tablet (Roche), and the membranes were collected by centrifugation at 40,000 g