| 1. |
- Generó, Magalí Martí, 1983-, et al.
(författare)
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A protocol for characterization of extremely preterm infant gut microbiota in double-blind clinical trials
- 2021
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Ingår i: STAR Protocols. - Cambridge, MA, United States : Cell press. - 2666-1667. ; 2:3
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Tidskriftsartikel (refereegranskat)abstract
- 16S rRNA gene sequencing enables microbial community profiling, but recovering fecal DNA from extremely premature infants is challenging. Here, we describe an optimized protocol for fecal DNA isolation, library preparation for 16S rRNA gene sequencing, taxonomy assignation, and statistical analyses. The protocol is complemented with a quantitative PCR for probiotic L. reuteri identification. This protocol describes how to characterize preterm infant gut microbiota and relate it to probiotic supplementation and clinical outcomes. It is customizable for other clinical trials. For complete details on the use and execution of this protocol, please refer to Martí et al. (2021) and Spreckels et al. (2021).
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| 2. |
- Generó, Magalí Martí, 1983-, et al.
(författare)
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Nitrogen and methanogen community composition within and among three Sphagnum dominated peatlands in Scandinavia
- 2015
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Ingår i: Soil Biology and Biochemistry. - : Elsevier BV. - 0038-0717 .- 1879-3428. ; 81, s. 204-211
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Tidskriftsartikel (refereegranskat)abstract
- Abstract Ombrotrophic raised bogs are nutrient poor acidic peatlands accumulating organic matter. They are widely spread on northern latitudes and are substantial sources of methane emissions to the atmosphere being of great concern from a climate change perspective. We investigated the methanogen community composition along microtopographic gradients within three bogs in Scandinavia, receiving different amounts of nitrogen precipitation. Methanogenic community analyses by terminal restriction fragment length polymorphism of the mcrA gene showed different profiles among the three sites, while no influence of the microtopographic gradients was observed. Peat temperature and dissolved organic carbon were the major edaphic variables explaining 38% of the variation of the methanogenic community diversity among the bogs. The family Methanoregulaceae (hydrogenotrophic methanogens) showed the largest relative proportion and highest activity in all three sites. Quantitative PCR of the mcrA gene and transcripts showed that the most northern site, receiving the lowest atmospheric nitrogen load, had significantly lower abundance and activity of methanogens (4.7 × 106 and 2.4 × 104 mcrA copies per gram of soil, respectively), compared to the most southern site (8.2 × 107 and 4.6 × 105 mcrA copies per gram of soil, respectively), receiving the highest nitrogen load. No patterns of the mcrA gene and transcript abundances were observed along the microtopography. The results indicated that the difference in occurrence of methanogens is mainly due to geoclimatological conditions rather than site intrinsic microtopographic variation. The study further suggests that environmental changes on the site intrinsic topography will not affect the methanogenic activity, while increasing average temperatures in Scandinavian ombrotrophic raised bogs might contribute to an increase of the methanogenic archaeal activity resulting in an increase of methane production.
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| 3. |
- Hellysaz, Arash, 1982-, et al.
(författare)
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Microbiota do not restrict rotavirus infection of colon
- 2023
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Ingår i: Journal of Virology. - : American Society for Microbiology. - 0022-538X .- 1098-5514. ; 97:11
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Tidskriftsartikel (refereegranskat)abstract
- Rotavirus is associated with extensive infection of the small intestine, whereas colon is considered to be uninfected. Considering that almost all bacteria in the gut colonize the colon, we hypothesized that the microbiota may act as a physical barrier preventing rotavirus infection in the colon in vivo. To address this hypothesis, we used human and mice colonoids, and biopsies of different intestinal segments of untreated and antibiotic-treated adult and infant mice. Rotavirus quantification was performed by qPCR and volumetric 3D imaging of intestinal segments. By 3D imaging, we observed infection in all the small intestinal segments, most extensively in the ileum, with most limited number of infected cells in colon. Broad-spectrum antibiotic treatment yielded no significant change in infection in either ileum or colon of adults and mice pups, although there is a substantial decrease in microbial load. We also show that rotavirus can successfully infect and replicate in colonoids from both mice and humans. Collectively, our data, including novel 3D imaging of the gut, mouse, and human colonoids, conclude that microbiota does not affect rotavirus infection in colon.IMPORTANCEAlterations of the gut microbiome can have significant effects on gastrointestinal homeostasis leading to various diseases and symptoms. Increased understanding of rotavirus infection in relation to the microbiota can provide better understanding on how microbiota can be used for clinical prevention as well as treatment strategies. Our volumetric 3D imaging data show that antibiotic treatment and its consequent reduction of the microbial load does not alter the extent of rotavirus infection of enterocytes in the small intestine and that restriction factors other than bacteria limit the infection of colonocytes.
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| 4. |
- Spreckels, Johanne, et al.
(författare)
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Lactobacillus reuteri Colonisation of Extremely Preterm Infants in a Randomised Placebo-Controlled Trial
- 2021
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Ingår i: Microorganisms. - : MDPI AG. - 2076-2607. ; 9:5
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Tidskriftsartikel (refereegranskat)abstract
- Lactobacillus reuteri DSM 17938 supplementation reduces morbidities in very low birth weight infants (<1500 g), while the effect on extremely low birth weight infants (ELBW, <1000 g) is still questioned. In a randomised placebo-controlled trial (ClinicalTrials.gov ID NCT01603368), head growth, but not feeding tolerance or morbidities, improved in L. reuteri-supplemented preterm ELBW infants. Here, we investigate colonisation with the probiotic strain in preterm ELBW infants who received L. reuteri DSM 17938 or a placebo from birth to postmenstrual week (PMW) 36. Quantitative PCR was used on 582 faecal DNA samples collected from 132 ELBW infants at one, two, three, and four weeks, at PMW 36, and at two years of age. Human milk oligosaccharides were measured in 31 milk samples at two weeks postpartum. At least 86% of the ELBW infants in the L. reuteri group were colonised with the probiotic strain during the neonatal period, despite low gestational age, high antibiotic pressure, and independent of infant feeding mode. Higher concentrations of lacto-N-tetraose, sialyl-lacto-N-neotetraose c, and 6 '-sialyllactose in mother's milk weakly correlated with lower L. reuteri abundance. Within the L. reuteri group, higher L. reuteri abundance weakly correlated with a shorter time to reach full enteral feeding. Female sex and L. reuteri colonisation improved head growth from birth to four weeks of age. In conclusion, L. reuteri DSM 17938 supplementation leads to successful colonisation in ELBW infants.
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| 5. |
- Wejryd, Erik, et al.
(författare)
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Low Diversity of Human Milk Oligosaccharides is Associated with Necrotising Enterocolitis in Extremely Low Birth Weight Infants
- 2018
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Ingår i: Nutrients. - : MDPI. - 2072-6643. ; 10:10
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Tidskriftsartikel (refereegranskat)abstract
- Difference in human milk oligosaccharides (HMO) composition in breast milk may be one explanation why some preterm infants develop necrotizing enterocolitis (NEC) despite being fed exclusively with breast milk. The aim of this study was to measure the concentration of 15 dominant HMOs in breast milk during the neonatal period and investigate how their levels correlated to NEC, sepsis, and growth in extremely low birth weight (ELBW; amp;lt;1000 g) infants who were exclusively fed with breast milk. Milk was collected from 91 mothers to 106 infants at 14 and 28 days and at postmenstrual week 36. The HMOs were analysed with high-performance anion-exchange chromatography with pulsed amperometric detection. The HMOs diversity and the levels of Lacto-N-difucohexaose I were lower in samples from mothers to NEC cases, as compared to non-NEC cases at all sampling time points. Lacto-N-difucohexaose I is only produced by secretor and Lewis positive mothers. There were also significant but inconsistent associations between 3-sialyllactose and 6-sialyllactose and culture-proven sepsis and significant, but weak correlations between several HMOs and growth rate. Our results suggest that the variation in HMO composition in breast milk may be an important factor explaining why exclusively breast milk fed ELBW infants develop NEC.
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