A volume of 25 mL of digesta from the colon was used for pH analysis using a digital pH meter (Dostmann electronic, GmbH, Wertheim, Germany) following the manufacturer’s protocol [82 ]. The ammonia concentration was measured using an ammonia selective-ion electrode (Orion 95–12, Orion Research Inc., Franklin, MA, USA) attached to a selective ion meter (710A model, Orion Research Inc., Franklin, MA, USA). The calibration was performed using dilutions at 10, 100, and 1000 mg/L of a 0.1 M standard ammonium chloride solution. In addition, an ISA solution (Ionic Strength Adjuster, Orion Research Inc., Franklin, MA, USA) of 0.5 mL was mixed into every 25 mL sample to ensure a uniform background ionic strength [83 (link)] The final results were reported as µmol/g.
Short-chain fatty acids from the piglet colonic digesta samples were analysed in aqueous extracts by gas chromatography as described by Marin et al. [80 (link)]. Briefly, the aqueous extracts were injected into a gas chromatograph (Varian, 430-GC, Varian Inc., Palo Alto, CA, USA) after centrifugation. The GC instrument is fitted with an Elite-FFAP capillary column (inside diameter 320 mm) (Perkin Elmer, Waltham, MA, USA). Volatile fatty acids obtained from CRM46975, Supelco, Bellefonte, PA, USA, were used as standards. The final results were reported as µmol/g.
Short-chain fatty acids from the piglet colonic digesta samples were analysed in aqueous extracts by gas chromatography as described by Marin et al. [80 (link)]. Briefly, the aqueous extracts were injected into a gas chromatograph (Varian, 430-GC, Varian Inc., Palo Alto, CA, USA) after centrifugation. The GC instrument is fitted with an Elite-FFAP capillary column (inside diameter 320 mm) (Perkin Elmer, Waltham, MA, USA). Volatile fatty acids obtained from CRM46975, Supelco, Bellefonte, PA, USA, were used as standards. The final results were reported as µmol/g.
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