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713 Lecture 15

713 Lecture 15. Host metagenomics. Progression of techniques. Culture based Use phenotypes and genotypes to ID Non-culture based, focused on 16S rDNA Clone 16S PCR products into vector and sequence Non-culture based, no cloning, sequence 16s

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713 Lecture 15

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  1. 713 Lecture 15 Host metagenomics

  2. Progression of techniques • Culture based • Use phenotypes and genotypes to ID • Non-culture based, focused on 16S rDNA • Clone 16S PCR products into vector and sequence • Non-culture based, no cloning, sequence 16s • Future: non-culture based, no cloning, sequence all genes being used (RT-PCR of RNA into cDNA, sequence)

  3. Some wild bugs detected in plaque

  4. Metagenomics • Metagenome – Analysis of the genetic material within a community of organisms Extract DNA Sequence DNA Analyze the types found

  5. More on the mammalian gut microbiome • Adult monozygotic twins (and, to a lesser extent, dizygotic twins and siblings) have more similar gut microbes than to their marital partners • Animals inherit their gut flora from their mothers • Host genotype appears to affect relative abundance of community members, not composition

  6. Mouse vs Fish • 99% of mouse gut bacteria belong to Bacteroides and Firmicutes • Mostly Proteobacteria in Fish • Do gut bacteria differ because the residents of their typical environment differ? • Or does the host animal select the resident bacteria?

  7. Responses of zebrafish to colonization with individual cultured gut microbes

  8. The bacteria must be fed to be helpful

  9. The Intestinal Microbiome • The gut microbiome is largely shared among family members • Your intestinal flora is closer to your mom and dad’s than your friend’s • In a study of 154 individuals, there was no common shared species between all individuals • There are 150 times more genes in the intestinal microbiome than in the human genome • Many of these bacteria perform metabolic functions necessary for digestion

  10. Co-Metabolism • The intestinal flora provide us with enzymes that can break down foods that the human genome does not encode • Glycoside hydrolase breaks down polysaccharides (such as pectin) • Our intestinal flora co-metabolizes our food and provides us with nutrients • Vitamin K, short chain fatty acids, biotin, Tryptophan, other amino acids, lipids, and indole-melanin • In a study involving rats, antibiotic intervention drastically altered the intermediate metabolites for many nutrients • After antibiotic treatment the rats were observed every day for the restoration of the metabolites to levels before antibiotic treatment

  11. Co-Metabolism • Antibiotics don’t just alter the microbial community • Provides proof in mammals that antibiotic treatment has effects on digestion • Reduced microbial flora changes metabolism

  12. Intestinal Flora and Obesity • Obese individuals have reduced microbial diversity • The amount of bacteroidetes decreases and firmicutes increase (associated with high fat diets) • Although bacteroidetes decrease, a subgroup of this phyla, Prevotellaceae, increases (No dietary restrictions) • The energy that microbes harvest from foods is increased in obese individuals compared to lean individuals • They release more calories, which in turn are taken up by the body

  13. Diet Changes the Intestinal Flora • A study using humanized mice with transplanted microbial communities from healthy individuals found that diet alters the intestinal community • The original community was heritable from generation to generation • When switched to a high fat diet, bacteroidetes decreased, firmicutes increased, and the mice started to gain weight • This new, alter community became transmissible from mother to offspring • This finding indicates that an unhealthy community can set up offspring for a less efficient microbial community

  14. Diet Changes the Intestinal Flora • A human study found that restricting calories and fat can change the microbial flora • Obese subjects were monitored for one year and were found to have Firmicutes and Bacteroidetes compose 92% of the intestinal flora • This trend was stable for all subjects for one year

  15. Diet Changes the Intestinal Flora • The subjects were then randomly assigned to fat restricted or carbohydrate restricted diets and monitored for another year

  16. The Intestinal Flora and Autoimmune Diseases • Not many direct links or causes have been made yet • It is hypothesized that properly regulated microbes send metabolic signals to the innate immune system • These signals can either suppress or stimulate the innate immune response

  17. The Intestinal Flora and Autoimmune Diseases • More studies are being conducted to elucidate the possible mechanistic causes of this microbe-immune system interaction • A trial treatment for Crohn’s disease and ulcerative colitis is fecal bacteriotherapy • Fecal transplant from healthy individuals into affected individuals • Shows promising results so far but needs clinical trials • Many people are “grossed out” by the treatment

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