Impact of Phlorotannin Extracts from Fucus vesiculosus on Human Gut Microbiota
Abstract
:1. Introduction
2. Results and Discussion
2.1. Stability, Bioaccessibility and Antioxidant Activity of F. vesiculosus Extracts throughout the Simulated GIT
2.2. Prebiotic Effect
2.3. Evolution of the Gut Microbiota Profile Groups
2.4. Organic Acids Profile and pH Variation
3. Materials and Methods
3.1. Chemicals
3.2. Extraction Procedure
3.3. Gastrointestinal Digestion Simulation
3.4. Determination of the Phlorotannin Content and Antioxidant Activities
3.5. Determination of the Phlorotannin Content and Antioxidant Activities
3.6. In Vitro Fermentation Assays
3.7. Gut Microbiota Evaluation
3.7.1. DNA Extraction
3.7.2. Real-Time PCR for Microbial Analysis of Stool
3.7.3. Determination of Organic Acids
3.8. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample | GIT Stage | TPhC (mg PGE/g ext) | (1) NO● (IC50 μg/mL) | (2) O2●– (IC50 μg/mL) |
---|---|---|---|---|
CRD | Undigested | 9.93 ± 1.48 a | 161 ± 8.8 a | 417 ± 164.5 a |
Mouth | 6.33 ± 2.96 b | 309 ± 105.2 b | 745 ± 88.2 b | |
Stomach | 8.52 ± 1.16 a,b | 171 ± 27.1 a | 378 ± 26.6 a | |
Intestine | 5.17 ± 0.70 b | 287 ± 27.2 a,b | 1105 ± 421.3 b | |
Retentate * | 4.60 ± 0.26 b | 141 ± 9.1 a | 294 ± 19.3 a | |
Permeate * | 1.40 ± 0.19 c | 2551 ± 30.7 c | 2580 ± 75.2 c | |
EtOAc | Undigested | 17.39 ± 1.77 a | 45 ± 2.5 a | 118 ± 17.6 a |
Mouth | 13.83 ± 0.74 b | 73 ± 11.0 a,b | 221 ± 1.1 a,b | |
Stomach | 5.67 ± 0.91 c | 109 ± 7.1 a,b | 244 ± 0.4 a,b | |
Intestine | 3.28 ± 0.55 c | 195 ± 38.5 b,c | 564 ± 19.9 c | |
Retentate * | 2.97 ± 0.62 c,d | 281 ±16.1 c | 383 ± 18.2 b,c | |
Permeate * | 0.37 ± 0.10 d | 1531 ± 52.2 d | 3074 ± 32.3 d | |
Standard compound | - | 36 ± 0.9 | 6 ± 0.5 |
Division (Genus) | Number of Copies (n = 5) a |
---|---|
Universal | 7.52 ± 0.38 |
Firmicutes | 4.76 ± 0.20 |
Clostridium leptum | 4.97 ± 0.26 |
Enterococcus spp. | 2.07 ± 0.63 |
Lactobacillus spp. | 3.27 ± 0.72 |
Bacteroidetes | 5.46 ± 0.63 |
Bacteroides spp. | 3.76 ± 0.55 |
Bifidobacterium spp. | 4.42 ± 0.45 |
F:B ratio | 0.97 ± 0.23 |
Organic Acids | Time (h) | Ctrl | FOS | CRD | EtOAc |
---|---|---|---|---|---|
Total | 0 | 2.38 ± 0.63 a;A | 2.38 ± 0.63 a;A | 2.38 ± 0.63 a;A | 2.38 ± 0.63 a;A |
12 | 5.24 ± 1.98 a;A | 10.89 ± 2.79 b;B | 7.43 ± 2.09 b;A | 7.76 ± 1.92 b;A,B | |
24 | 4.90 ± 1.59 a;A | 12.63 ± 2.37 b,c;B | 7.08 ± 2.45 b;A | 7.55 ± 1.75 b;A | |
48 | 4.10 ± 2.01 a;A | 14.78 ± 4.00 c;B | 5.04 ± 1.57 a,b;A | 6.38 ± 1.98 b;A | |
Succinic acid | 0 | 0.45 ± 0.20 a;A | 0.45 ± 0.20 a;A | 0.45 ± 0.20 a;A | 0.45 ± 0.20 a;A |
12 | 0.77 ± 0.75 a;A | 1.85 ± 0.92 b;B | 2.29 ± 1.39 c;B | 2.15 ± 1.15 b;B | |
24 | 1.12 ± 0.53 a;A | 1.97 ± 0.58 b;B | 2.02 ± 0.93 b,c;A,B | 1.35 ± 0.50 a,b;A,B | |
48 | 0.74 ± 0.71 a;A | 2.03 ± 0.85 b;B | 1.12 ± 0.20 a,b;A,B | 1.40 ± 0.89 a,b;A,B | |
Lactic acid | 0 | ND | ND | ND | ND |
12 | 1.21 ± 0.93 a;A | 3.91 ± 1.94 a;B | 0.87 ± 0.22 a;A | 0.87 ± 0.23 a;A | |
24 | 0.34 ± 0.14 a;A | 4.81 ± 0.75 a,b;B | 0.76 ± 0.58 a;A | 0.26 ± 0.16 a;A | |
48 | ND | 5.49 ± 2.14 b | ND | ND | |
Acetic acid | 0 | 0.16 ± 0.04 a;A | 0.16 ± 0.04 a;A | 0.16 ± 0.04 a;A | 0.16 ± 0.04 a;A |
12 | 0.81 ± 0.10 b;A | 1.36 ± 0.75 b;A | 1.03 ±0.09 b;A | 1.02 ± 0.19 b;A | |
24 | 0.82 ± 0.17 b;A | 1.65 ± 0.52 b;B | 0.92 ± 0.22 b;A | 0.96 ± 0.30 b;A | |
48 | 0.78 ± 0.20 b;A | 2.77 ± 1.21 c;B | 0.78 ± 0.20 b;A | 0.93 ± 0.28 b;A | |
Propionic acid | 0 | 0.34 ± 0.09 a;A | 0.34 ± 0.09 a;A | 0.34 ± 0.09 a;A | 0.34 ± 0.09 a;A |
12 | 0.53 ± 0.23 a;A | 1.48 ± 0.32 b;B | 1.14 ± 0.49 b;B | 1.43 ± 0.87 b;B | |
24 | 0.65 ± 0.35 a;A | 1.89 ± 0.75 b;C | 1.25 ± 0.58 b;B | 0.85 ± 0.27 a,b;A,B | |
48 | 0.50 ± 0.24 a;A | 1.64 ± 0.60 b;B | 0.77 ± 0.20 a,b;A | 0.90 ± 0.24 a,b;A | |
Butyric acid | 0 | 1.41 ± 0.25 a;A | 1.41 ± 0.25 a;A | 1.41 ± 0.25 a;A | 1.41 ± 0.25 a;A |
12 | 1.92 ± 0.69 a;A | 2.29 ± 0.99 a;A | 2.10 ± 0.79 a;A | 2.71 ± 0.94 a;A | |
24 | 2.24 ± 0.67 a;A | 2.23 ± 0.86 a;A | 2.54 ± 1.05 a;A | 4.12 ± 0.37 b;B | |
48 | 2.23 ± 1.35 a;A | 2.70 ± 1.43 a;A | 2.31 ± 0.85 a;A | 4.31 ± 0.62 b;B |
Target Group | Maximum Growth Rate (µmax.h−1) | |||
---|---|---|---|---|
Primer Sequence (5′–3′) | Genomic DNA Standard | PCR Product Size (bp) | AT (°C) | |
Universal | AAA CTC AAA GGA ATT GAC GG ACT TCA CGA GCT GAC | Bacteroides vulgatus ATCC 8482 (DSMZ 1447) | 180 | 45 |
Firmicutes | ATG TGG TTT AAT TCG AAG CA AGC TGA CGA CAA CCA TGC AC | Lactobacillus gasseri ATCC 33323 (DSMZ 20243) | 126 | 45 |
Enterococcus spp. | CCC TTA TTG TTA GTT GCC ATC ATT ACT CGT TGT ACT TCC CT TGT | Enterococcus gilvus ATCC BAA-350 (DSMZ 15689) | 144 | 45 |
Lactobacillus spp. | GAG GCA GCA GTA GGG AAT CTT C GGC CAG TTA CTA CCT CTA TCC TTC TTC | Lactobacillus gasseri ATCC 33323 (DSMZ 20243) | 126 | 55 |
Bacteroidetes | CAT GTG GTT TAA TTC GAT GAT AGC TGA CGA CAA CCA TGC AG | Bacteroides vulgatus ATCC 8482 (DSMZ 1447) | 126 | 45 |
Bacteroides spp. | ATA GCC TTT CGA AAG RAA GAT CCA GTA TCA ACT GCA ATT TTA | Bacteroides vulgatus ATCC 8482 (DSMZ 1447) | 495 | 45 |
Bifidobacterium spp. | CGC GTC TGG TGT GAA AG CCC CAC ATC CAG CAT CCA | Bifidobacterium longum subsp. infantis ATCC 15697 (DSMZ 20088) | 244 | 50 |
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Catarino, M.D.; Marçal, C.; Bonifácio-Lopes, T.; Campos, D.; Mateus, N.; Silva, A.M.S.; Pintado, M.M.; Cardoso, S.M. Impact of Phlorotannin Extracts from Fucus vesiculosus on Human Gut Microbiota. Mar. Drugs 2021, 19, 375. https://doi.org/10.3390/md19070375
Catarino MD, Marçal C, Bonifácio-Lopes T, Campos D, Mateus N, Silva AMS, Pintado MM, Cardoso SM. Impact of Phlorotannin Extracts from Fucus vesiculosus on Human Gut Microbiota. Marine Drugs. 2021; 19(7):375. https://doi.org/10.3390/md19070375
Chicago/Turabian StyleCatarino, Marcelo D., Catarina Marçal, Teresa Bonifácio-Lopes, Débora Campos, Nuno Mateus, Artur M. S. Silva, Maria Manuela Pintado, and Susana M. Cardoso. 2021. "Impact of Phlorotannin Extracts from Fucus vesiculosus on Human Gut Microbiota" Marine Drugs 19, no. 7: 375. https://doi.org/10.3390/md19070375