Blueberry Husks and Probiotics Attenuate Colorectal Inflammation and Oncogenesis, and Liver Injuries in Rats Exposed to Cycling DSS-Treatment.

Long-term colonic inflammation promotes carcinogenesis and histological abnormalities of the liver, and colorectal tumours frequently arise in a background of dysplasia, a precursor of adenomas. Altered colonic microbiota with an increased proportion of bacteria with pro-inflammatory characteristics, have been implicated in neoplastic progression. The composition of the microbiota can be modified by dietary components such as probiotics, polyphenols and dietary fibres. In the present study, the influence of probiotics in combination with blueberry husks on colorectal carcinogenesis and subsequent liver damage was evaluated.Colorectal tumours were induced in rats by cyclic treatment with dextran sulphate sodium (DSS). Blueberry husks and a mixture of three probiotic strains (Bifidobacterium infantis DSM 15159, Lactobacillus gasseri, DSM 16737 and Lactobacillus plantarum DSM 15313) supplemented a basic diet fortified with oats. The condition of the rats was monitored using a disease activity index (DAI). A qualitative and quantitative histological judgement was performed on segments of distal colon and rectum and the caudate lobe of the liver. The formation of short-chain fatty acids, bacterial translocation, the inflammatory reaction and viable count of lactobacilli and Enterobaceriaceae were addressed.Blueberry husks with or without probiotics significantly decreased DAI, and significantly reduced the number of colonic ulcers and dysplastic lesions. With a decreased proportion of blueberry husk in the diet, the probiotic supplement was needed to achieve a significant decrease in numbers of dysplastic lesions. Probiotics decreased faecal viable count of Enterobacteriaceae and increased that of lactobacilli. Blueberry husks with or without probiotics lowered the proportion of butyric acid in distal colon, and decreased the haptoglobin levels. Probiotics mitigated hepatic injuries by decreasing parenchymal infiltration and the incidence of stasis and translocation. The results demonstrate a dietary option for use of blueberry husks and probiotics to delay colonic carcinogenesis and hepatic injuries in the rat model

Colorectal Oncogenesis and Inflammation in a Rat Model Based on Chronic Inflammation due to Cycling DSS Treatments

Inflammation is known to be linked with development of colorectal cancer, and the aim was to assess the malignant potential and degree of inflammation in a dextran-sulphate-sodium-(DSS-) induced cyclic colonic tumour model (CTM) in rats and to compare it with the azoxymethane-(AOM-) induced CTM model. Tumours developed in both groups, although, in the DSS group, the colonic mucosa appeared edematous and the number of haemorrhagic erosions and quantity of dysplastic lesions were higher as well as the mucosal concentration of myeloperoxidase and faecal viable count of Enterobacteriaceae. The livers were affected as evaluated by steatosis, parenchymal loss, haemorrhage, and inflammatory infiltrations, and higher proportions of acetate and lower proportions of butyrate in colonic content were found. The DSS model seems to mimic the clinical situation and may be valuable for investigation of inflammation-related dysplasia and colon cancer, as well as for altered liver function by endogenous inflammatory mediators

Intestinal function, microflora and nutrient intake of children after administration of a fermented oat product containing Lactobacillus plantarum DSM 9843 (299v)

The objective of the study was to evaluate the intestinal tolerance of a newly developed food containing Lactobacillus plantarum 299v, in children. The effects of consumption of the product on the children's nutrient intake were also followed, in a separate study. The tolerance study was parallel, double-blind, and included 69 children between 6 months and 3 years of age, randomized into two groups. One of the groups consumed a fermented oat product (1×109 cfu L. plantarum 299v/g) for 3 weeks, while the other group consumed a placebo product. We analysed the subjects by intention to treat and also carried out an analysis of subjects who had a mean intake of >100 g study product per day. After consumption for 3 weeks, the content of L. plantarum 299v in faeces increased significantly in the test group compared with the placebo group (p<0.001) and mean log10 cfu/g was 8.7 (intention to treat analysis). Also, the total content of lactobacilli increased and was significantly higher compared with the placebo group (p<0.001). The same result was also obtained when the data for the subjects who had a higher consumption of the study products were analysed. There were no differences in reported gastrointestinal function between the groups. The effects of the fermented oat product on the children's nutrient intake were followed in 12 children aged 1-3 years. Inclusion of the fermented oat product that was enriched with iron and ascorbic acid led to a significantly higher intake of several nutrients like energy, carbohydrates (g, E%), dietary fibre, iron and zinc. In conclusion, the children tolerated the fermented oat product well, the faecal microflora was positively altered and the children's nutritional intake was improved. Keywords: children, microflora, Lactobacillus plantarum 299v, probiotic bacteria, intestinal tolerance, nutrient intake

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

Evaluation of liver injury (Incidence).

<p>Liver specimens were histologically evaluated for the incidence of steatosis, infiltrating inflammatory cells in steatotic and non-steatotic areas, vascular stasis and loss of liver parenchyma. The status of the livers of different groups is expressed as incidence of phenomena.</p>1<p>Cellinfiltration around CV within steatotic areas.</p>2<p>Cellinfiltration elsewhere in the parenchyma.</p><p>Incidence: * denotes P<0.05 compared with the C group.</p

Incidence of translocations and identified isolates from the livers.

<p>Thirty-four isolates were subjected to 16 S rDNA sequencing and the similarity levels for each isolate are shown in the table. Translocation is mentioned as incidence of phenomena/total number of animals. Isolates were received from Rogosa agar/Brain Heart Infusion agar (incubated anaerobically)/Brain Heart Infusion agar (incubated aerobically). Between brackets are species with the same sequence similarity as the before mentioned.</p><p>Incidence: * denotes P<0.05 compared with the C group.</p><p><i>Lactobacillus animalis</i> (<i>L. animalis</i>), <i>Lactobacillus apodemi</i> (<i>L. apodemi</i>), <i>Kocuria rhizophila</i> (<i>K. rhizophila</i>), <i>Lactobacillis frumenti</i> (<i>L. frumenti</i>), <i>Lactobacillus antri</i> (<i>L. antri</i>), <i>Lactobacillus gasseri</i> (<i>L. gasseri</i>), <i>Micrococcus luteus</i> (<i>M. luteus</i>), <i>Clostridium ramosum</i> (<i>C. ramosum</i>), <i>Staphylococcus warneri</i> (<i>S. warneri</i>), <i>Leifsonia xyli</i> subsp. <i>cynodontis</i> (<i>L. xyli subsp. cynodontis</i>), <i>Enterococcus casseliflavus</i> (<i>E. casseliflavus</i>), <i>Clostridium perfringens</i> (<i>C. perfringens</i>), <i>Paenibacillus lautus</i> (<i>P. lautus</i>), <i>Kocuria rosea</i> (<i>K. rosea</i>), <i>Corynebacterium lipophiloflavum</i> (<i>C. lipophiloflavum</i>), <i>Clostridium subterminale</i> (<i>C. subterminale</i>), <i>Bacillus siralis</i> (<i>B. siralis</i>).</p

Disease activity index.

<p>Disease activity index (DAI) during the 11 cycles of DSS administration. DAI scores are expressed as medians (25 and 75 percentiles). Significant differences are expressed versus the C group. Cycle 1: **P<0.01 for groups 2B, 2BP and B; Cycle 2: **P<0.01 for groups B and BP, *** P<0.001 for groups 2B and 2BP groups; Cycle 3: *P<0.05 for group P, **P<0.001 for group B group, ***P<0.001 for groups 2B, 2BP and BP; Cycle 4: *P<0.05 for group BP, ***P<0.001 for groups 2B and 2BP; Cycle 5: **P<0.01 for group B group, ***P<0.001 for groups 2B, 2BP, and BP; Cycle 6: *P<0.05 for group B, **P<0.01 for group BP,***P<0.001 for groups 2B and BP; Cycle 7: *P<0.05 for group B, **P<0.01 for groups 2B and B, ***P<0.001 for groups 2BP and BP; Cycle 8: *P<0.05 for group BP, **P<0.01 for groups 2B and B, ***P<0.001 for group 2BP; Cycle 9: *P<0.05 for groups 2B, 2BP, B, BP; Cycle 10, *P<0.05 for group B, **P<0.01 for groups 2B and 2BP; Cycle 11: *P<0.05 for groups 2B, 2BP, B, BP.</p

SCFAs in colon content and blood samples.

<p>Proportions (%) of acetic-, propionic- and butyric acids in the hindgut and blood of rats given diets supplemented with different amounts of blueberry husks (2B = 122 g blueberry/kg dwb and B = 61 g blueberry/kg dwb) and/or probiotics (P; 6⋅10⁹ CFU per day).</p><p>Probiotics: <i>Bifidobacterium infantis</i> (CURE21), <i>Lactobacillus gasseri</i> VPG44, and <i>Lactobacillus plantarum</i> (HEAL19).</p><p>Mean values for eight rats per group, B group (n = 6).</p><p>Mean values of C, 2B and B only, i.e. those without any added probiotics, with unlike superscript letters were significantly different (P<0.05).</p><p>Mean values were significantly different from those rats fed diets without bacteria: *P<0.05, ** P<0.01, *** P<0.001.</p>1<p>2B (n = 7), B and BP (n = 6).</p