Impact of Novel Sorghum Bran Diets on DSS-Induced Colitis.

We have demonstrated that polyphenol-rich sorghum bran diets alter fecal microbiota; however, little is known regarding their effect on colon inflammation. Our aim was to characterize the effect of sorghum bran diets on intestinal homeostasis during dextran sodium sulfate (DSS)-induced colitis. Male Sprague-Dawley rats (N = 20/diet) were provided diets containing 6% fiber from cellulose, or Black (3-deoxyanthocyanins), Sumac (condensed tannins) or Hi Tannin Black (both) sorghum bran. Colitis was induced (N = 10/diet) with three separate 48-h exposures to 3% DSS, and feces were collected. On Day 82, animals were euthanized and the colon resected. Only discrete mucosal lesions, with no diarrhea or bloody stools, were observed in DSS rats. Only bran diets upregulated proliferation and Tff3, Tgfß and short chain fatty acids (SCFA) transporter expression after a DSS challenge. DSS did not significantly affect fecal SCFA concentrations. Bran diets alone upregulated repair mechanisms and SCFA transporter expression, which suggests these polyphenol-rich sorghum brans may suppress some consequences of colitis.

Space Environmental Factor Impacts upon Murine Colon Microbiota and Mucosal Homeostasis.

Astronaut intestinal health may be impacted by microgravity, radiation, and diet. The aim of this study was to characterize how high and low linear energy transfer (LET) radiation, microgravity, and elevated dietary iron affect colon microbiota (determined by 16S rDNA pyrosequencing) and colon function. Three independent experiments were conducted to achieve these goals: 1) fractionated low LET γ radiation (137Cs, 3 Gy, RAD), high Fe diet (IRON) (650 mg/kg diet), and a combination of low LET γ radiation and high Fe diet (IRON+RAD) in male Sprague-Dawley rats; 2) high LET 38Si particle exposure (0.050 Gy), 1/6 G partial weight bearing (PWB), and a combination of high LET38Si particle exposure and PWB in female BalbC/ByJ mice; and 3) 13 d spaceflight in female C57BL/6 mice. Low LET radiation, IRON and spaceflight increased Bacteroidetes and decreased Firmicutes. RAD and IRON+RAD increased Lactobacillales and lowered Clostridiales compared to the control (CON) and IRON treatments. Low LET radiation, IRON, and spaceflight did not significantly affect diversity or richness, or elevate pathogenic genera. Spaceflight increased Clostridiales and decreased Lactobacillales, and similar trends were observed in the experiment using a ground-based model of microgravity, suggesting altered gravity may affect colonic microbiota. Although we noted no differences in colon epithelial injury or inflammation, spaceflight elevated TGFß gene expression. Microbiota and mucosal characterization in these models is a first step in understanding the impact of the space environment on intestinal health.

Polyphenol-rich sorghum brans alter colon microbiota and impact species diversity and species richness after multiple bouts of dextran sodium sulfate-induced colitis.

The microbiota affects host health, and dysbiosis is involved in colitis. Sorghum bran influences butyrate concentrations during dextran sodium sulfate (DSS) colitis, suggesting microbiota changes. We aimed to characterize the microbiota during colitis, and ascertain if polyphenol-rich sorghum bran diets mitigate these effects. Rats (n = 80) were fed diets containing 6% fiber from cellulose, or Black (3-deoxyanthocyanins), Sumac (condensed tannins), or Hi Tannin black (both) sorghum bran. Inflammation was induced three times using 3% DSS for 48 h (40 rats, 2 week separation), and the microbiota characterized by pyrosequencing. The Firmicutes/Bacteroidetes ratio was higher in Cellulose DSS rats. Colonic injury negatively correlated with Firmicutes, Actinobacteria, Lactobacillales and Lactobacillus, and positively correlated with Unknown/Unclassified. Post DSS#2, richness was significantly lower in Sumac and Hi Tannin black. Post DSS#3 Bacteroidales, Bacteroides, Clostridiales, Lactobacillales and Lactobacillus were reduced, with no Clostridium identified. Diet significantly affected Bacteroidales, Bacteroides, Clostridiales and Lactobacillus post DSS#2 and #3. Post DSS#3 diet significantly affected all genus, including Bacteroides and Lactobacillus, and diversity and richness increased. Sumac and Hi Tannin black DSS had significantly higher richness compared to controls. Thus, these sorghum brans may protect against alterations observed during colitis including reduced microbial diversity and richness, and dysbiosis of Firmicutes/Bacteroidetes.

Increased dietary iron and radiation in rats promote oxidative stress, induce localized and systemic immune system responses, and alter colon mucosal environment.

Astronauts are exposed to increased body iron stores and radiation, both of which can cause oxidative damage leading to negative health effects. The purpose of this study was to investigate combined effects of high dietary iron (650 mg/kg diet) and radiation exposure (0.375 Gy cesium-137 every other day for 16 d) on markers of oxidative stress, immune system function, and colon mucosal environment in male Sprague-Dawley rats (n=8/group). Control rats consumed adequate iron (45 mg/kg diet) and were not irradiated. Combined treatments increased liver glutathione peroxidase, serum catalase, and colon myeloperoxidase while decreasing total fecal short-chain fatty acid concentrations. The high-iron diet alone increased leukocyte count. Radiation decreased the T-cell CD4:CD8 ratio. Plasma iron was negatively correlated with cytokine production in activated monocytes. Genes involved in colon microbial signaling, immune response, and injury repair were altered by radiation. Genes involved with injury repair and pathogen recognition changed with dietary iron. These data demonstrate that dietary iron and radiation, alone and combined, contribute to oxidative stress that is related to immune system alterations in circulation and the colon. The model presented may help us better understand the changes to these systems that have been identified among astronauts.

Characterization of fecal microbiota in cats using universal 16S rRNA gene and group-specific primers for Lactobacillus and Bifidobacterium spp.

The diversity of the feline intestinal microbiota has not been well elucidated. The aim of this study was to characterize fecal microbiota of cats by comparative sequence analysis with universal bacterial 16S rRNA gene and group-specific primers for Bifidobacterium and Lactobacillus spp. Using universal bacterial primers, a total of 133 non-redundant 16S rRNA gene sequences were identified in fecal samples obtained from 15 healthy pet cats. The majority of these sequences were assigned to the phylum Firmicutes, followed by Proteobacteria and Bacteroidetes. Further classification showed that Firmicutes were predominantly affiliated with Clostridium clusters XI, XIVa, and I. Using group-specific primers for Bifidobacterium and Lactobacillus spp., 364 clones were analyzed in fecal samples obtained from 12 additional cats and these bacterial genera were observed in 100% and 92% of cats, respectively. These detection rates differed from those obtained using universal bacterial primers, where Bifidobacterium and Lactobacillus spp. were each detected in 2 cats (13.3%). Overall, 23 different Lactobacillus-like and 11 Bifidobacterium-like sequences were identified. We observed marked differences in the prevalence of the various lactic acid bacteria in individual cats. In conclusion, the use of a combination of universal and group-specific primers allows a more detailed characterization of lactic acid bacteria in the feline intestine. While Bifidobacterium and Lactobacillus spp. are prevalent in feline fecal samples, individual animals show a unique species distribution.

Assessment of microbial diversity along the feline intestinal tract using 16S rRNA gene analysis.

The aim of this study was to describe the microbial communities along the gastrointestinal tract in healthy cats based on analysis of the 16S rRNA gene. Gastrointestinal content (i.e. content from the stomach, duodenum, jejunum, ileum, and colon) was collected from four healthy conventionally raised colony cats and one healthy specific pathogen-free (SPF) cat. Bacterial 16S rRNA genes were amplified using universal bacterial primers and analyzed by comparative sequence analysis. A total of 1008 clones were analyzed and 109 nonredundant 16S rRNA gene sequences were identified. In the four conventionally raised cats, five different bacterial phyla were observed, with sequences predominantly classified in the phylum Firmicutes (68%), followed by Proteobacteria (14%), Bacteroidetes (10%), Fusobacteria (5%), and Actinobacteria (4%). The majority of clones fell within the order Clostridiales (54%), followed by Lactobacillales, Bacteroidales, Campylobacterales, and Fusobacteriales (14%, 11%, 10%, and 6%, respectively). Clostridiales were predominantly affiliated with Clostridium clusters I (58%) and XIVa (27%). The intestinal microbiota of the SPF cat displayed a reduced bacterial diversity, with 98% of all clones classified in the phylum Firmicutes. Further classification showed that the Firmicutes clones belonged exclusively to the class Clostridiales and were predominantly affiliated with Clostridium cluster I.