Impact of functional flours from pineapple by-products on human intestinal microbiota

Impact of Phlorotannin Extracts from Fucus vesiculosus on Human Gut Microbiota

“…The numbers were in agreement with those found in healthy volunteers’ feces, with Clostridium , Bacteroides and Bifidobacterium comprising the dominant genera while Lactobacillus spp. and Enterococcus appeared as the subdominant genera [ 31 , 32 , 33 ]. Figure 2 depicts the relative differences (in %) between the microbiota groups of the tested samples and control feces, along 12, 24 and 48 h of fermentation.…”

“…The simulation of the gastrointestinal digestion of the F. vesiculosus sample extracts was performed according to the method described by Campos et al [ 33 ]. Oral digestion was started by suspending 1 g of dried sample (CRD or EtOAc) in 20 mL of distilled water followed by the adjustment of the pH between 5.6 and 6.9 with NaHCO 3 prior to the addition of 0.6 mL/min of α-amylase at 100 U/mL.…”

“…Moreover, the donors confirmed that they were not taking any probiotic or prebiotic supplements, as well as any form of antibiotics for the previous 3 months. The basal medium was prepared as described previously [ 33 ], consisting of a nutrient base medium containing 5.0 g/L trypticase soya broth (TSB) without dextrose (BBL, Cockeysville, Maryland, MD, USA), 5.0 g/L bactopeptone (Amersham, Buckinghamshire, UK), 0.5 g/L L-cysteine-HCl (Merck, Germany), 1.0% ( v / v ) of salt solution A (100.0 g/L NH 4 Cl, 10.0 g/L MgCl 2 6H 2 O, 10.0 g/L CaCl 2 2H 2 O), 0.2% ( v / v ) of salt solution B (200.0 g/L K 2 HPO 4 3H 2 O) and 0.2% ( v / v ) of 0.5 g/L resazurin solution, prepared in distilled water and with pH adjustment at 6.8. The basal medium was dispensed into airtight glass anaerobic bottles, sealed with aluminum caps before sterilization by autoclave.…”

“…Genomic DNA was extracted and purified from stool samples as previously described [ 33 ] using NZY Tissue gDNA Isolation Kit (Nzytech, Lisbon, Portugal) with some modifications. Samples were centrifuged at 11,000 g during 10 min to separate the supernatant from the pellet.…”

“…Real-time PCR was performed as described before in [ 33 ] in sealed 96-well microplates using a LightCycler FastStart DNA Master SYBR Green kit and a LightCycler instrument (Roche Applied Science, Indianapolis, ID, USA). PCR reaction mixtures (total of 10 μL) contained 5 μL of 2 × Faststart SYBRGreen (Roche Diagnostics Ltd., Burgess Hill, UK), 0.2 μL of each primer (final concentration of 0.2 μM), 3.6 μL of water and 1 μL of DNA (equilibrated to 20 mg).…”