Probiotics and berry-associated polyphenols: catabolism and antioxidative effects

Oxidative stress can cause damage to DNA, proteins and lipids and is associated with inflammation and various human diseases as cancer, atherosclerosis, and autoimmune diseases. Polyphenol-rich diet, such as fruits and berries, may act as antioxidants and prevent oxidative stress and, thereby, associated diseases. Administration of lactic acid bacteria (LAB) can affect the microflora in the gastrointestinal (GI) tract and may increase the capability of the bacterial flora to digest polyphenols. Some strains of Lactobacillus may break down phenolic acids and hydrolyzable tannins into phenolic metabolites that are more easily absorbed in the body and may enhance antioxidative effects. The aim of this thesis was to clarify the protective effects of polyphenol-rich fruits and berries alone or in combination with different strains of LAB on oxidative stress in mice. Furthermore, transformation of polyphenols in a bilberry beverage by LAB was examined. Supplementation with rosehips of the rose species Rosa pimpinellifolia or an LAB mixture decreased lipid peroxidation and oxidative stress in colon of mice after ischemia-reperfusion (I/R) injury. Adding an LAB supplement to the rosehips increased the concentrations of phenolic compounds, antioxidative capacity and total phenolic content in cecum. Rosehips of R. piminellifolia are a rich source of cyanidin-3-O-glucoside and this compound and its degradation product, protocatechuic acid, were detected in the cecum content. Administration of bilberry, either alone or together with Lactobacillus plantarum HEAL19, decreased lipid peroxidation and oxidative stress in colon of mice after I/R injury. A chokeberry-supplement showed no antioxidative effect. Bilberry was found to have a more complex anthocyanin profile than chokeberry. Higher concentrations and a more varied composition of anthocyanins were seen in colon than in cecum. More phenolic metabolites were found in the intestines of bilberry-fed mice than in the chokeberry-fed ones. Chokeberry or bilberry alone decreased the number of LAB on the colonic mucosa but addition of L. plantarum HEAL19 prevented this reduction. In a more extensive ischemia-reperfusion injury, diet supplemented with bilberry, but without addition of different LAB strains, reduced lipid peroxidation and protected the small intestine against oxidative stress. The highest concentration and recovery of anthocyanins was seen in the ileal content followed by that of colon and finally cecum. Anthocyanin arabinosides, and especially malvidin-3-O-arabinoside, were accumulated in the colon content. Glucosides and galactosides of malvidin, peonidin and petunidin seemed to be digested by the microflora in the cecum. Supplementation of bilberry to the diet influenced the composition of cecum microflora. Anthocyanins in bilberry beverages inoculated with different LAB strains, alone or in combination with wine yeast, decreased during 3 weeks incubation at 30 degrees Celsius. Arabinosides of malvidin and petunidin showed the greatest decrease. Addition of yeast improved the stability of the anthocyanins. In contrast to anthocyanins, quercetin, quercetin-3-glucoside and detected phenolic acids were relatively stable. Antioxidative capacity and total phenolic content decreased in all samples. In conclusion, dietary supplementation of rosehips from Rosa pimpinellifolia or bilberry suppressed oxidative stress in colonic tissue of mice. Protective effects may be due to the high anthocyanin content, presence of phenolic metabolites and changed microflora. Addition of LAB improved status of the colonic but not the ileal tissue

Transfer of gut microbiota from lean and obese mice to antibiotic-treated mice

Transferring gut microbiota from one individual to another may enable researchers to “humanize” the gut of animal models and transfer phenotypes between species. To date, most studies of gut microbiota transfer are performed in germ-free mice. In the studies presented, it was tested whether an antibiotic treatment approach could be used instead. C57BL/6 mice were treated with ampicillin prior to inoculation at weaning or eight weeks of age with gut microbiota from lean or obese donors. The gut microbiota and clinical parameters of the recipients was characterized one and six weeks after inoculation. The results demonstrate, that the donor gut microbiota was introduced, established, and changed the gut microbiota of the recipients. Six weeks after inoculation, the differences persisted, however alteration of the gut microbiota occurred with time within the groups. The clinical parameters of the donor phenotype were partly transmissible from obese to lean mice, in particularly β cell hyperactivity in the obese recipients. Thus, a successful inoculation of gut microbiota was not age dependent in order for the microbes to colonize, and transferring different microbial compositions to conventional antibiotic-treated mice was possible at least for a time period during which the microbiota may permanently modulate important host functions

Treatment with a Monoclonal Anti-IL-12p40 Antibody Induces Substantial Gut Microbiota Changes in an Experimental Colitis Model

Background and Aim. Crohn’s disease is associated with gut microbiota (GM) dysbiosis. Treatment with the anti-IL-12p40 monoclonal antibody (12p40-mAb) has therapeutic effect in Crohn’s disease patients. This study addresses whether a 12p40-mAb treatment influences gut microbiota (GM) composition in mice with adoptive transfer colitis (AdTr-colitis). Methods. AdTr-colitis mice were treated with 12p40-mAb or rat-IgG2a or NaCl from days 21 to 47. Disease was monitored by changes in body weight, stool, endoscopic and histopathology scores, immunohistochemistry, and colonic cytokine/chemokine profiles. GM was characterized through DGGE and 16S rRNA gene-amplicon high-throughput sequencing. Results. Following 12p40-mAb treatment, most clinical and pathological parameters associated with colitis were either reduced or absent. GM was shifted towards a higher Firmicutes-to-Bacteroidetes ratio compared to rat-IgG2a treated mice. Significant correlations between 17 bacterial genera and biological markers were found. The relative abundances of the RF32 order (Alphaproteobacteria) and Akkermansia muciniphila were positively correlated with damaged histopathology and colonic inflammation. Conclusions. Shifts in GM distribution were observed with clinical response to 12p40-mAb treatment, whereas specific GM members correlated with colitis symptoms. Our study implicates that specific changes in GM may be connected with positive clinical outcomes and suggests preventing or correcting GM dysbiosis as a treatment goal in inflammatory bowel disease

Antioxidative protection of dietary bilberry, chokeberry and Lactobacillus plantarum HEAL19 in mice subjected to intestinal oxidative stress by ischemia-reperfusion

<p>Abstract</p> <p>Background</p> <p>Ischemia-reperfusion (I/R) in the intestines is an inflammatory condition which activates leukocytes and reactive oxygen species (ROS) and leads to lipid peroxidation and DNA damage. Bilberry and chokeberry fruits are rich sources of polyphenols which may act as antioxidants and prevent lipid peroxidation. Lactic acid bacteria (LAB) may improve microbial status in the intestines and increase the metabolic activity towards polyphenolic degradation. The aim of the study was to clarify antioxidative effects of bilberry and chokeberry fruits alone and with addition of a LAB-strain, <it>Lactobacillus plantarum </it>HEAL19, in an I/R-model in mice.</p> <p>Methods</p> <p>Male BALB/cJ mice were fed the experimental diets for 10 days. Diets consisted of standard chow supplemented with either bilberry (<it>Vaccinium myrtillus</it>) or chokeberry (<it>Aronia × prunifolia</it>) powder alone or in combination with the LAB-strain <it>Lactobacillus plantarum </it>HEAL19. I/R-injury was induced by holding superior mesenteric artery clamped for 30 minutes followed by reperfusion for 240 minutes. Thereafter, colonic and caecal tissues and contents were collected. Malondialdehyde (MDA) was used as indicator of lipid peroxidation and was measured by a calorimetric assay, lactobacilli were cultured on Rogosa agar plates and <it>Enterobacteriaceae </it>on VRBG agar plates, anthocyanins and phenolic acids were analysed by HPLC-DAD-ESI-MSn.</p> <p>Results</p> <p>MDA was significantly decreased in the colon of groups fed bilberry alone (p = 0.030) and in combination with <it>L. plantarum </it>HEAL19 (p = 0.021) compared to the IR-control but not in chokeberry-fed groups. Supplementation with bilberry or chokeberry alone reduced the total number of lactobacilli on the mucosa. Higher concentrations of anthocyanins were found in the colon than in the caecum content of mice. A more varied composition of different anthocyanins was also observed in the colon content compared to the caecum of bilberry-fed mice. Phenolic acids formed by microbial degradation of the dietary polyphenols in the gut could be detected. More phenolic metabolites were found in the intestines of bilberry-fed mice than in the chokeberry-fed ones.</p> <p>Conclusions</p> <p>Bilberry alone and in combination with <it>L. plantarum </it>HEAL19 exerts a better protection against lipid peroxidation than chokeberry. These dietary supplements may be used to prevent or suppress oxidative stress.</p

Effects of Bilberry (Vaccinium myrtillus) in Combination with Lactic Acid Bacteria on Intestinal Oxidative Stress Induced by Ischemia-Reperfusion in Mouse.

Intestinal ischemia-reperfusion (I/R) results in oxidative stress, inflammation, and tissue injuries. The present study investigates the antioxidative and anti-inflammatory effects of a dietary supplement of bilberry, either alone or in combination with Lactobacillus plantarum RESO56, L. plantarum HEAL19, or Pediococcus acidilactici JAM046, in an I/R-induced model for oxidative stress in mice. A bilberry diet without addition of bacteria significantly decreased both lipid peroxidation (p = 0.001) and mucosal injury in the ileum. Of 14 anthocyanins identified in bilberry, anthocyanin arabinosides were the most resistant to absorption and microbial degradation in the intestines. Cyanidin-3-glucoside and delphinidin-3-glucoside seemed to be mostly absorbed in the stomach and upper part of the small intestine, while malvidin-3-galactoside, peonidin-3-glucoside, peonidin-3-galactoside, and petunidin-3-galactoside seemed to be digested by the microbiota in the cecum. Bilberry strongly influenced the composition of the cecal microbiota. In conclusion, a food supplement of bilberry protected small intestine against oxidative stress and inflammation induced by ischemia-reperfusion

Effects on weight gain and gut microbiota in rats given bacterial supplements and a high-energy-dense diet from fetal life through to 6 months of age

The aim of this study was to assess the long-term effects of a high-energy dense diet, supplemented with Lactobacillus plantarum (Lp) or Escherichia coli (Ec) on weight gain, fattening and the gut microbiota in rats. Since the mother’s dietary habits can influence offspring physiology, the dietary regimes started with the dams at pregnancy and through lactation, and continued with the offspring for six months. The weight gain of group Lp was lower than for groups C (control) and Ec (P=0•086). More retroperitoneal adipose tissue (P=0•030) and higher plasma leptin (P=0•035) were seen in group Ec compared to group Lp. The viable count of Enterobacteriaceae was higher in group Ec than in group Lp (P=0•019) and when all animals were compared, Enterobacteriaceae correlated positively with body weight (r=0•428, P=0•029). Bacterial diversity was lower in group Ec than in groups C (P=<0•05) and Lp (P=<0•05). Firmicutes, Bacteroidetes and Verrucomicrobia dominated in all groups, but Bacteroidetes were more prevalent in group C than in groups Lp (P=0•036) and Ec (P=0•056). The same five bacterial families dominated the microbiota of groups Ec and C, and four of these were also present in group Lp. The other five families dominating in group Lp were not found in any of the other groups. Multivariate data analysis pointed in the same directions as the univariate statistics. Our results suggest that supplementation of L. plantarum or E. coli can have long-term effects on the composition of the intestinal microbiota, as well as on weight gain and fattening