27 research outputs found

    The Gut Bacterial Flora - Focus on Early Life and Physiological Traits

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    The gastrointestinal tract of the foetus is considered sterile but during vaginal birth the neonate comes into contact with bacteria from the maternal vaginal and intestinal microbiota. The main focus of this doctoral thesis was to elucidate the initial bacterial ecosystem in newborns and to relate microbial perturbations to physiological traits. The bacterial flora was mainly studied with molecular-genetic methods. When the microbiota was assessed in one-week-old infants, reduced faecal bacterial diversity was found in infants developing atopic eczema at the age of 18 months compared to those infants not developing eczema. To further elucidate the pioneer microbiota, stool samples from healthy full-term vaginally-born neonates was studied. Lactobacillus was found in all newborns within 48 hours after birth. Species commonly found in the vaginal microbiota were to some extent detected among the babies. Other bacterial groups were found in varying prevalence. Interestingly, a subgroup of neonates born large for gestational age had significantly more Proteobacteria compared to neonates born appropriate for gestational age. Maternal microbiota and dietary habits are known to impact offspring physiology. Results presented in this thesis show impaired physiology in suckling rat pups to dams of the outbred Sprague-Dawley stock when these were treated with high-energy dense diet during the gestation and lactation period. Offspring body weight, adiposity, gut permeability and systemic inflammation were further accentuated if the Gram-negative Escherichia coli was given to the dams in combination with the high-energy dense feed. In a similar experimental design, the pups were monitored in a longitudinal study. E. coli exposure from foetal life until six months of age decreased the diversity of the caecal microbiota along with enhanced adiposity. In contrast, when the Gram-positive Lactobacillus plantarum was given instead of E. coli, body weight gain and fat accumulation were lower in addition to a more favourable gut microbiota, implying the prospect of effect on health homeostasis by bacterial consumption. This thesis suggests that a high load of E. coli should be avoided in pregnant mothers and in children, and treatment with L. plantarum may be a therapeutic option. However, more extensive research is needed to establish the relationship between inflammation, obesity and the bacterial flora of the gut. A general conclusion concerning the microbiota is that the intestinal ecosystem should be studied at least at the hierarchical level of genus or family, but preferably on the species level since looking at the phylum level can give superficial information

    Buserelin treatment to rats causes enteric neurodegeneration with moderate effects on CRF-immunoreactive neurons and Enterobacteriaceae in colon, and in acetylcholine-mediated permeability in ileum

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    The gonadotropin-releasing hormone (GnRH) analog buserelin causes enteric neuronal loss. Acute stress or injection of corticotropin-releasing factor (CRF) affects motility, secretion, and barrier function of the gastrointestinal tract. The aim of the study was to characterize the CRF immunoreactivity in enteric neurons after buserelin treatment, and to evaluate possible effects of enteric neuropathy on gut microbiota, intestinal permeability, and stress response behavior

    Pre-treatment with antibiotics and Escherichia coli to equalize the gut microbiota in conventional mice.

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    The composition of the gut microbiota can vary widely between individual mice of the same batch and thereby affect the resulting outcome in experimental studies. Therefore, an efficient method is needed to equalize the gut microbiota prior to the start of critical experiments. In order to minimize variations in gut microbiota between animals and provide the animals with a Gram-negative flora exposing lipopolysaccharides in the cell-walls, C57BL/6 mice were given a mixture of ampicillin, metronidazole and clindamycin in the drinking water for 3 days and then Escherichia coli for two additional days. Treatment with antibiotics alone or with antibiotics in combination with E. coli was well tolerated by all animals. Body weight and liver weight were not affected, although higher hepatic fat content was found in treated animals (p < 0.05). The diversity of the gut microbiota was strongly reduced in animals treated with antibiotics and antibiotics in combination with E. coli (p < 0.01), without affecting the total amount of bacteria. Cloned and sequenced 16S rRNA genes showed high presence of Enterobacteriaceae and Porphymonadaceae in the treated animals. Analysis with Principal Component Analysis gave a clear separation of the composition in microbiota between different treatment groups. The described treatment efficiently equalized the gut microbiota and provided the animals with a strong abundance of Enterobacteriaceae without changing the total load of bacteria. This is a straightforward, lenient and efficient method of pre-treatment to equalize the gut microbiota of mice as a starting procedure of animal studies

    Probiotic therapy to men with incipient arteriosclerosis initiates increased bacterial diversity in colon: A randomized controlled trial.

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    OBJECTIVE: This study aimed to clarify the microbial change in the intestinal microbiota in patients, with cardiovascular disease, consuming a drink with high numbers of live Lactobacillus plantarum. METHODS: Sixteen males, with atherosclerotic plaque on the carotid wall, were randomly selected from a larger cohort and included in this double blind, placebo controlled study. Colonic biopsies, taken before and after four weeks of probiotic treatment, were analysed with Terminal Restriction Fragment Length Polymorphism, including digestion with MspI and HaeIII. Microbial diversity was calculated, short-chain fatty acids in faeces, and blood markers were analysed. RESULTS: Consumption of one probiotic strain of L. plantarum (DSM 9843) increased intestinal microbial diversity. The probiotic group had an increased diversity after consumption of the probiotic drink compared to the change in the placebo group when Shannon and Weaner diversity index (MspI and HaeIII, p=0.026) and Simpson index of diversity (MspI, p=0.044 and HaeIII, p=0.026) were calculated. The fermentation pattern of short-chain fatty acids in faeces were unaffected for most acids, but the probiotic group had decreased concentration of isovaleric acid (p=0.006) and valeric acid (p=0.029). Viable count of lactobacilli increased in the probiotic group (p=0.001), but no significant changes in blood markers were observed. CONCLUSION: Administration of a single-strain probiotic increases the bacterial diversity in the gut, and affects the concentration of some short-chain fatty acids. Consumption of the single strain L. plantarum DSM 9843 might be a strategy to favour a diverse intestinal microbiota, which is beneficial for the host

    New Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus strains : well tolerated and improve infant microbiota

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    Background: Different microorganisms from the environment will begin to colonise the infant during and immediately after the delivery. It could be advantageous to influence the microbiome early on by giving infants probiotic bacteria. The aim of the study was to investigate the tolerance of two probiotic lactobacilli in infants. The effect on the microbiota was also followed. Methods: Thirty-six healthy infants, aged 4–83 days at the start of the study, were given a daily supplementation of probiotics (Lactiplantibacillus plantarum HEAL9 and Lacticaseibacillus rhamnosus 271, 109 CFU (colony-forming units)) or placebo for 8 weeks. Adverse events, growth parameters, the faecal microbiome and intestinal performance were followed. Results: No differences between the groups in growth parameters, adverse events and intestinal performance were observed. The faecal levels of L. plantarum, L. rhamnosus and lactobacilli increased after the intake of probiotics and were significantly higher compared with the placebo group after 4 and 8 weeks of intake. The faecal microbial diversity was similar in the two groups at the end of the study. Conclusions: The intervention with the probiotic formulation was well tolerated and increased the level of lactobacilli in the intestine. The developed probiotic formulation will be further evaluated for clinical efficacy in infants. Impact: New data for the development of the gut function and the microbiome in breastfed and/or formula-fed young infants over time and the effect of adding two probiotic strains are presented.Lactiplantibacillusplantarum is a species that seldom has been analysed in infants, but it could be detected in 25% of the subjects before administration (mean age 41 days).Lactiplantibacillusplantarum and L. rhamnosus establish well in the intestine of infants and are well tolerated.The microbiota was positively affected by the intake of probiotics

    Dietary green-plant thylakoids decrease gastric emptying and gut transit, promote changes in the gut microbial flora, but does not cause steatorrhea

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    Green-plant thylakoids increase satiety by affecting appetite hormones such as ghrelin, cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1). The objective of this study was to investigate if thylakoids also affect gastrointestinal (GI) passage and microbial composition. To analyse the effects on GI passage, 16 rats were gavage-fed a control or thylakoid-supplemented high-fat diet (HFD) 30 min before receiving Evans blue. Another 16 rats were fed a control HFD or thylakoid HFD for two weeks prior to the intragastric challenge with Evans blue. The amount of Evans blue in the stomach and the distance of migration in the intestines after 30 min were used as a measurement of gastric emptying and intestinal transit. These were reduced by thylakoid supplementation in the acute study, and however not significantly also after the two-week diet study. The second aim of the study was to investigate if thylakoid-supplementation affects the gut microbiota and amount of faecal fat in healthy human volunteers (n = 34) receiving thylakoid or placebo treatments for three months. Microbiota was analysed using 16S rRNA gene sequencing and qPCR, and faecal fat was extracted by dichloromethane. The total bacteria, and specifically the Bacteriodes fragilis group, were increased by thylakoid treatment versus placebo, while thylakoids did not cause steatorrhea. Dietary supplementation with thylakoids thus affects satiety both via appetite hormones and GI fullness, and affects the microbial composition without causing GI adverse effects such as steatorrhea. This suggests thylakoids as a novel agent in prevention and treatment of obesity

    Anthropometric and metabolic improvements in human type 2 diabetes after introduction of an Okinawan-based Nordic diet are not associated with changes in microbial diversity or SCFA concentrations

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    The Okinawan-based Nordic (O-BN) diet improves anthropometry and metabolism in type 2 diabetes mellitus (T2DM) patients. The aim of this study was to study mechanisms behind improvements by examining Enterobacteriaceae abundance, microbial diversity, and concentrations of short-chain fatty acids (SCFAs). A secondary aim was exploring if metformin treatment affects microbiota or SCFAs. Thirty T2DM patients received the O-BN diet for 12 weeks. Faecal and blood samples were collected at baseline, 12 and 28 weeks. Although patients experienced weight loss and improved metabolic parameters, there were no significant changes in Enterobacteriaceae abundance or microbial diversity. Patients on metformin displayed higher Enterobacteriaceae abundance throughout the study (p = .008, p = .038, and p = .001, respectively). Isovaleric acid was decreased after 12 weeks (p = .018). Butyric acid was decreased at follow-up (p = .007). Improved anthropometry and metabolism in T2DM after introduction of the O-BN diet is not associated with changes in Enterobacteriaceae abundance, microbial diversity or SCFA concentrations

    Abundance of Enterobacteriaceae in the colon mucosa in diverticular disease

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    AIM:To compare gut bacterial diversity and amount of Enterobacteriaceae in colonic mucosa between patients with and without diverticular disease (DD).METHODS:Patients in a stable clinical condition with planned elective colonoscopy were included. Blood samples and colon mucosa biopsies were collected at the colonoscopy. Study questionnaires including questions about gastrointestinal symptoms were completed by the patients and physicians. DNA from mucosa samples was isolated and the amount of Enterobacteriaceae was estimated using PCR assay. Terminal restriction fragment length polymorphism was applied to assess microbial diversity. Diversity was estimated by calculations of richness (number of terminal restriction fragments) and Shannon-Wiener and Simpson's indices.RESULTS:A total of 51 patients were included, 16 patients with DD [68 (62-76) years] and 35 controls [62 (40-74) years] without any diverticula. Patients with DD had significantly higher levels of Enterobacteriaceae than those without DD (P = 0.043), and there was an inverse relationship between the amount of Enterobacteriaceae and the Simpson's index (rs = -0.361, P = 0.033) and the Shannon-Wiener index (rs = -0.299, P = 0.081). The Simpson's index (P = 0.383), Shannon-Wiener index (P = 0.401) or number of restrictions fragments (P = 0.776) did not differ between DD and controls. The majority of patients experienced gastrointestinal symptoms, and 22 patients (43.1%) fulfilled the criteria for irritable bowel syndrome, with no difference between the groups (P = 0.212). Demography, socioeconomic status, lifestyle habits, inflammatory biomarkers, or symptoms were not related to the amount of Enterobacteriaceae or bacterial diversity.CONCLUSION:Patients with DD had higher amount of Enterobacteriaceae in the colon mucosa compared to patients without diverticula
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