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Probiotics should bring ‘balance’ to the intestinal microbiota by stimulating beneficial bacteria, whilst mitigating adverse ones. Balance can also be interpreted as high alpha-diversity. Contrary, Escherichia coli is often regarded as an adverse component of the resident intestinal microbiota. The aim of the present study was to implement a mouse model for in vivo screening of Lactobacillus-strains for ability to increase gut-microbiota diversity and to mitigate E. coli. Mice were divided into six groups, two dietary control-groups and four groups administered strains of Lactobacillus fermentum and/or Lactobacillus plantarum. All animals were pre-treated with antibiotics, and E. coli in order to equalise the microbiota from the start. After 7 weeks of Lactobacillus administration, the animals were sacrificed: DNA was extracted from caecum tissue, and the microbiota composition was analysed with terminal restriction fragment length polymorphism (T-RFLP) and 16S rRNA gene sequencing. The diversity of the caecal microbiota decreased when the dietary carbohydrate source was limited to corn starch. Conversely, the diversity was restored by Lactobacillus-supplements. The tested combinations of two Lactobacillus strains exerted different influences, not only on the taxonomic level, but also on the inferred microbiome functions. The mixture of L. fermentum GOS47 and L. fermentum GOS1 showed potential for anti-inflammatory activity and short chain fatty acid production. On the other hand, co-administration of L. fermentum GOS57 and L. plantarum GOS42 significantly decreased the viable count of Enterobacteriaceae. These results warrant further investigation of the tested strains as candidates for probiotics. Furthermore, the findings demonstrated that the current experimental animal model is suitable for in vivo studies of the effect of bacterial supplements on the gut-microbiota.

Background Oral administration of health-promoting bacteria is increasingly used in clinical practise. These bacteria have anti-inflammatory characteristics and modulate the immune system without major reported side effects. The mechanisms of action are not yet fully defined. Our aim was to study systemic effects of probiotics by measurements of leukocytes as well as local effects on rectal mucosal biopsies after adding a standardized inflammatory stimulus in vitro. Methods Fourteen healthy subjects were randomized to receive 1010 colony forming units/day orally of the probiotic strain Lactiplantibacillus plantarum 299 (Lp299), n = 7, or Bifidobacterium infantis CURE21 (CURE21), n = 7, for six weeks. Rectal biopsies were taken before and after ingestion of either probiotic strain product, for stimulation in vitro with tumour necrosis factor alpha (TNF-α) at 10 and 100 ng/ml respectively up to 8 h. Blood tests were sampled before and after treatment. Lactate dehydrogenase (LDH) confirmed viable tissue. Results Composition of the intestinal microbiota was not changed. Systemic leukocytes decreased after administration of CURE21 (PP Conclusions Administration of these probiotic strains to healthy humans show both a systemic and local reduction of inflammatory response by lowering leukocyte counts, and for Lp299 IL-6 levels in rectal mucosa. Probiotics may play an important role in the reduction of inflammatory responses expected after trauma during surgery or after pelvic irradiation. Trial registration Clinical Trials, registration number NCT01534572, retrospectively registered (http://www.clinicaltrials.gov).

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.

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.

It is common practice in India to consume the dairy drink buttermilk as a way of mitigating occupational heat strain. This paper explores the thermoregulatory and hydration benefits of drinking buttermilk but also the impacts of work in a hot environment on the gut microbiota, renal and cognitive function. Twelve healthy participants were subjected to a 3-h period of medium load physical intermittent work in a climatic chamber (34°C, 60% RH). The subjects were given water, buttermilk (700 ml) or no rehydration at random. Mean body temperatures when no rehydration was given were significantly higher (p≤0.001). When subjects drank water or buttermilk they had a lower sweat rate than with no rehydration (p≤0.05) and the perception of feeling hot, uncomfortable, thirsty and physically exerted was significantly reduced (p≤0.05). A hormonal stress response at the end of the exposure was seen when not drinking (p≤0.05). No differences in cognitive abilities and gut microbiota were found. The exposure lowered the renal blood flow suggesting an acute impact of short term heat exposure. It was also found that buttermilk has a protective effect on this impact. Our results demonstrated that keeping hydrated by water/buttermilk consumption mitigates heat strain in well-nourished subjects.

The virtual version of the Trier Social Stress Test (V-TSST) is an effective and standardized tool for social stress induction. This study aimed to examine gut permeability and physiological and inflammatory markers of reactivity to acute psychosocial stress. Forty young men were classified as high-stressed (HIGHS) or low-stressed (LOWS) according to the Shirom-Melamed Burnout Questionnaire. Cardiovascular reactivity and gut dysfunction were studied along with cortisol, zonulin and cytokines. Gut permeability was shown to be affected within one hour after the psychosocial stress induction, and shown to be dependent on age. Interleukin-6 increased with time, most pronounced at the end of the one-hour recovery after V-TSST, and was positively correlated to age. HIGHS experienced more abdominal dysfunction compared to LOWS. In conclusion, this study is the first to show fluctuations in gut permeability after psychosocial stress induction. This was partly associated with changes in inflammatory markers.

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.

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.

Barley husks, rye bran, and a fiber residue from oat milk production were processed by heat pretreatment, various separation steps, and treatment with an endoxylanase in order to improve the prebiotic potential of these cereal byproducts. Metabolic functions were intended to improve along with improved microbial activity. The products obtained were included in a high-fat mouse diet so that all diets contained 5% dietary fiber. In addition, high-fat and low-fat controls as well as partially hydrolyzed guar gum were included in the study. The soluble fiber product obtained from rye bran caused a significant increase in the bifidobacteria (log copies of 16S rRNA genes; median (25-75 percentile): 6.38 (6.04-6.66) and 7.47 (7.30-7.74), respectively; p < 0.001) in parallel with a tendency of increased production of propionic acid and indications of improved metabolic function compared with high-fat fed control mice. The oat-derived product caused an increase in the pool of cecal propionic (from 0.62 ± 0.12 to 0.94 ± 0.08) and butyric acid (from 0.38 ± 0.04 to 0.60 ± 0.04) compared with the high-fat control, and it caused a significant increase in lactobacilli (log copies of 16S rRNA genes; median (25-75 percentile): 6.83 (6.65-7.53) and 8.04 (7.86-8.33), respectively; p < 0.01) in the cecal mucosa. However, no changes in measured metabolic parameters were observed by either oat or barley products.

Antibiotics are frequently administered orally to treat bacterial infections not necessarily related to the gastrointestinal system. This has adverse effects on the commensal gut microbial community, as it disrupts the intricate balance between specific bacterial groups within this ecosystem, potentially leading to dysbiosis. We hypothesized that modulation of community composition and function induced by antibiotics affects intestinal integrity depending on the antibiotic administered. To address this a total of 60 Wistar rats (housed in pairs with 6 cages per group) were dosed by oral gavage with either amoxicillin (AMX), cefotaxime (CTX), vancomycin (VAN), metronidazole (MTZ), or water (CON) daily for 10-11 days. Bacterial composition, alpha diversity and caecum short chain fatty acid levels were significantly affected by AMX, CTX and VAN, and varied among antibiotic treatments. A general decrease in diversity and an increase in the relative abundance of Proteobacteria was observed for all three antibiotics. Additionally, the relative abundance of Bifidobacteriaceae was increased in the CTX group and both Lactobacillaceae and Verrucomicrobiaceae were increased in the VAN group compared to the CON group. No changes in microbiota composition or function were observed following MTZ treatment. Intestinal permeability to 4 kDa FITC-dextran decreased after CTX and VAN treatment and increased following MTZ treatment. Plasma haptoglobin levels were increased by both AMX and CTX but no changes in expression of host tight junction genes were found in any treatment group. A strong correlation between the level of caecal succinate, the relative abundance of Clostridiaceae 1 family in the caecum, and the level of acute phase protein haptoglobin in blood plasma was observed. In conclusion, antibiotic-induced changes in microbiota may be linked to alterations in intestinal permeability, although the specific interactions remain to be elucidated as changes in permeability did not always result from major changes in microbiota and vice versa.

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

Background 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. Results Sixty rats were given buserelin (20 μg) or saline subcutaneously for 5 days, repeated four times with 3 weeks in-between. At the study end, enteric neuronal density, enteric expression of CRF, gut microbial composition, and plasma levels of adrenocorticotropic hormone (ACTH) and CRF were analyzed. Intestinal permeability was examined in Ussing chambers and the reaction to stressful events was measured by behavior tests. Buserelin treatment reduced the number of neurons along the entire gastrointestinal tract, with increased relative numbers of CRF-immunoreactive submucosal and myenteric neurons in colon (p

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.