Influence of Human Milk on Very Preterms’ Gut Microbiota and Alkaline Phosphatase Activity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design and Participants
2.2. Enteral Nutrition Protocol
2.3. Clinical Data Collection
2.4. Fecal Sample Collection and DNA Extraction
2.5. Quantitative Analysis of Fecal Microbiota by RT-PCR
2.6. Microbial 16S rRNA Sequence Analysis
2.7. Alkaline Phosphatase Activity Assay
2.8. Statistical Analysis
3. Results
3.1. Clinical Characterization of the Preterm Neonates
3.2. Impact of Feeding on Intestinal Bacterial Establishment in Premature Neonates
3.3. Gut Microbiota Profile at the 26th Postnatal Day According to Feeding Types
3.4. Influence of Infant Feeding on Fecal Alkaline Phosphatase Activity
3.5. Impact of MOM in Different Proportions
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target Group | Primer Sequence (5′-3′) | Genomic DNA Standard | AT | Ref. |
---|---|---|---|---|
Total bacteria | AAA CTC AAA KGA ATT GAC GG CTC ACR RCA CGA GCT GAC | Bacteroides vulgatus ATCC 8482 | 62 °C | [22] |
Bacteroidetes | CAT GTG GTT TAA TTC GAT GAT AGC TGA CGA CAA CCA TGC AG | Bacteroides vulgatus ATCC 8482 | 60 °C | [16] |
Firmicutes | ATG TGG TTT AAT TCG AAG CA AGC TGA CGA CAA CCA TGC AC | Lactobacillus gasseri ATCC 33323 | 60 °C | [16] |
γ-Proteobacteria | TCGTCAGCTCGTGTYGTGA CGTAAGGGCCATGATG | E. coli ATCC 25922 | 61 °C | [22] |
Lactobacillus | GAG GCA GCA GTA GGG AAT CTT C GGC CAG TTA CTA CCT CTA TCC TTC TTC | Lactobacillus gasseri ATCC 33323 | 60 °C | [16] |
Bifidobacterium | CGC GTC YGG TGT GAA AG CCC CAC ATC CAG CAT CCA | Bifidobacterium longum ATCC 15697 | 60 °C | [16] |
Bacteroides | ATA GCC TTT CGA AAG RAA GAT CCA GTA TCA ACT GCA ATT TTA | Bacteroides vulgatus ATCC 33563 | 60 °C | [16] |
Enterococcus | CCC TTA TTG TTA GTT GCC ATC ATT ACT CGT TGT ACT TCC CT TGT | Enterococcus gilvus ATCC BAA-350 | 61 °C | [16] |
Escherichia coli | GTA AGT TAC ACT ATA AAA GCA CCG TCG TCT GTG TGG ATG GTA ATA AAT TTT TG | Escherichia coli ATCC 25922 | 60 °C | [23] |
MOM (n = 75) | DHM (n = 20) | Formula (n = 13) | p-Value (MOM vs. DHM) | p-Value (MOM vs. Formula) | p-Value (DHM vs. Formula) | |
---|---|---|---|---|---|---|
Extremely/very preterm n | 28/47 | 5/15 | 0/13 | 0.205 | 0.004 | 0.065 |
Gestational age (mean ± SD) | 28.3 ± 2.1 | 28.6 ± 1.8 | 29.6 ±1.1 | 0.103 | 0.003 | 0.17 |
Vaginal delivery/C-section, n | 34/41 | 7/13 | 1/12 | 0.407 | 0.010 | 0.074 |
Somatometry at birth (mean ± SD) | ||||||
Weight, g | 1123 ± 345 | 1173 ± 284 | 1416 ± 219 | 0.438 | 0.002 | 0.011 |
Length, cm | 36.2 ± 3.3 | 36.8 ± 3.2 | 39.1 ± 2.0 | 0.428 | 0.003 | 0.027 |
Cephalic perimeter, cm | 25.4 ± 2.4 | 25.3 ± 1.8 | 27.2 ± 1.5 | 0.996 | 0.007 | 0.008 |
Z-score | ||||||
Weight at birth | −0.27 ± 0.82 | 0.04 ± 1.45 | 0.15 ± 0.70 | 0.977 | 0.057 | 0.789 |
Length at birth | −0.44 ± 1.09 | −0.25 ± 1.61 | 0.04 ±0.65 | 0.777 | 0.051 | 0.543 |
Cephalic perimeter at birth | −0.46 ± 0.90 | −0.68 ± 1.79 | −0.77 ± 1.28 | 0.731 | 0.147 | 0.331 |
Weight at 26th | −0.08 ± 1.85 | 0.85 ± 1.62 | 1.65 ± 1.13 | 0.257 | 0.001 | 0.132 |
Δ weight until 26th day, g | 200 ± 190 | 200 ± 170 | 420 ± 160 | 0.671 | <0.001 | 0.001 |
Postnatal day of full enteral nutrition (mean ± SD) | 16.5 ± 8.5 | 16.4 ± 5.6 | 9.5 ± 4.2 | 0.401 | 0.002 | <0.001 |
Days of antibiotherapy (mean ± SD) | 12 ± 11 | 13 ± 8 | 5 ± 3 | 0.178 | 0.010 | <0.001 |
Days of hospitalization (mean ± SD) | 58 ± 24 | 59 ± 21 | 42 ± 12 | 0.874 | 0.026 | 0.018 |
MOM | DHM | Formula | |||
---|---|---|---|---|---|
95% CI | p-Value | 95% CI | p-Value | ||
Total Bacteria | |||||
Gestational age-adjusted | 1 (referent) | −0.556 (−1.093 to −0.079) | 0.023 | −0.889 (−1.341 to −0.438) | <0.001 |
Mode of delivery-adjusted | 1 (referent) | −0.565 (−1.036 to −0.095) | 0.018 | −0.552 (−1.047 to −0.057) | 0.029 |
Multivariable-adjusted a | 1 (referent) | −0.659 (–1.082 to −0.236) | 0.002 | −0.721 (−1.158 to −0.284) | 0.001 |
Firmicutes | |||||
Gestational age-adjusted | 1 (referent) | −0.585 (−1.183 to 0.013) | 0.055 | −0.789 (−1.322 to −0.256) | 0.004 |
Mode of delivery-adjusted | 1 (referent) | −0.587 (−1.161 to −0.013) | 0.045 | −0.456 (−1.060 to 0.162) | 0.139 |
Multivariable-adjusted a | 1 (referent) | −0.610 (–1.114 to −0.107) | 0.017 | −0.759 (−1.279 to −0.238) | 0.004 |
Bifidobacterium | |||||
Gestational age-adjusted | 1 (referent) | −2.002 (−3.315 to −0.689) | 0.003 | −2.092 (−3.262 to −0.922) | <0.001 |
Mode of delivery-adjusted | 1 (referent) | −0.291 (−1.016 to 0.434) | 0.431 | −0.684 (−1.447 to 0.079) | 0.079 |
Multivariable-adjusted a | 1 (referent) | −0.656 (−1.388 to 0.075) | 0.079 | −0.975 (−1.716 to −0.233) | 0.010 |
* | <90% to ≥80% | <80% to ≥70% | <70% to ≥50% | <50% to ≥35% | <35% | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|
95% CI | p | 95% CI | p | 95% CI | p | 95% CI | p | 95% CI | p | ||
Chao1 index | |||||||||||
Multivariable -adjusted a | Ref. | 2.559 (–0.707 to 5.824) | 0.125 | −2.384 (–5.749 to 0.980) | 0.165 | 1.603 (–2.467 to 5.673) | 0.440 | −9.451 (−15.660 to−3.242) | 0.003 | −2.820 (–6.273 to 0.634) | 0.110 |
Total Bacteria | |||||||||||
Multivariable -adjusted a | Ref. | −0.173 (–0.697 to 0.351) | 0.518 | −0.179 (–0.684 to 0.326) | 0.487 | 0.104 (–0.355 to 0.562) | 0.658 | −0.611 (–1.235 to 0.12) | 0.055 | −0.771 (–1.311 to −0.231) | 0.005 |
Firmicutes | |||||||||||
Multivariable -adjusted a | Ref. | −0.551 (–1.165 to 0.063) | 0.079 | −0.502 (–1.094 to 0.090) | 0.097 | −0.401 (–0.939 to 0.137) | 0.144 | −1.297 (–2.028 to −0.566) | 0.001 | −1.115 (–1.749 to −0.482) | 0.001 |
Bifidobacterium | |||||||||||
Multivariable -adjusted a | Ref. | 0.605 (−0.495 to 1.704) | 0.281 | −0.039 (−1.018 to 1.096) | 0.943 | −0.237 (−1.196 to 0.723) | 0.628 | 0.207 (−1.078 to 1.492) | 0.753 | −0.424 (−1.543 to 0.696) | 0.458 |
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Morais, J.; Marques, C.; Faria, A.; Teixeira, D.; Barreiros-Mota, I.; Durão, C.; Araújo, J.; Ismael, S.; Brito, S.; Cardoso, M.; et al. Influence of Human Milk on Very Preterms’ Gut Microbiota and Alkaline Phosphatase Activity. Nutrients 2021, 13, 1564. https://doi.org/10.3390/nu13051564
Morais J, Marques C, Faria A, Teixeira D, Barreiros-Mota I, Durão C, Araújo J, Ismael S, Brito S, Cardoso M, et al. Influence of Human Milk on Very Preterms’ Gut Microbiota and Alkaline Phosphatase Activity. Nutrients. 2021; 13(5):1564. https://doi.org/10.3390/nu13051564
Chicago/Turabian StyleMorais, Juliana, Cláudia Marques, Ana Faria, Diana Teixeira, Inês Barreiros-Mota, Catarina Durão, João Araújo, Shámila Ismael, Sara Brito, Manuela Cardoso, and et al. 2021. "Influence of Human Milk on Very Preterms’ Gut Microbiota and Alkaline Phosphatase Activity" Nutrients 13, no. 5: 1564. https://doi.org/10.3390/nu13051564