Topic 7 - Diet, the gut microbiome, & human health (PART 1) Flashcards
Define microbiome
the collective genomes of the micro-organisms in a particular environment
The first meaning is an inclusive one that refers to an entire habitat where microorganisms dwell, and encompasses the microbes, their genomes, and the surrounding environment.
The second, narrower, meaning is “the collection of genes and genomes of members of a microbiota.”
Define microbiota
the community of micro-organisms themselves
Define microbiota diversity
a measure of how many different species, and dependent on the diversity indices, how evenly distributed they are in the community
lower diversity is considered a marker of dysbiosis (microbial imbalance) in the gut and has been found in autoimmune diseases & obesity and cardiometabolic conditions, as well as in elderly people
Define colonocytes
epithelial cells of the colon
Define germ-free animals
animals that have no micro-organisms living in or on them
Define short chain fatty acids
fatty acids with 2 or 6 carbon atoms that are produced by bacterial fermentation of dietary fibres
How were microbiomes studied in the ‘olden’ days?
- microscopes
- culture dependent methods (pure culture)
problem is that not everything can be cultured
What are the main challenges of new sequencing techniques used to study microbiomes?
- hard to distinguish them via morphology
- hard to grow
or is this culture-dependent technique challenges??? I think it might be
DNA to understand the taxonomy of microbes
- conserved regions are agreed upon to use as a barcode
- must be conserved for all microbes but different enough to tell species apart
- established protocols to extract DNA, amplify DNA, and determine barcode
What is the V4 region of ribosomes?
- the region in purple is the area that is amplified by DNA primers
- it is agreed upon as a way to taxonomically distinguish bacterial species
Why can’t the V4 region be used to classify fungi?
Because one of the differences b/w bacteria & eukaryotes (like fungi) is that we have different ribosomes & thus diff ribosomal subunits
a different subunit would have to be amplified for the fungi
How does sequencing occur?
- targeted regions are amplified (bioinformatic treatment)
- merge 2 together
- then we cluster them together based on similarity (OTU)
- when we put that through a data base to assign taxonomy, we can determine which species each sample belongs to
- the abundance can also be determined
What is alpha diversity?
a measure of species richness (number of taxa) within a single microbial environment
- how many different microbial species could be detected in a specific sample
What is beta diversity?
a measure of diversity in microbial community b/w different environments (difference in taxonomic abundance profiles from diff samples)
- how different is the microbial composition in one environment compared to another
How can we work out what the gut microbes are doing?
- diff techniques to amplify the DNA and assemble into WHOLE genomes (shotgun sequencing)
- gives the whole genome, and can link this to the microbiome
- can understand a pathway which is being expressed when RNA is used
“metagenome”
What are the 3 basic functions of microbes?
1) digest food components
2) produce vitamins
3) stimulate the immune system
If we transplant a stool sample from an obese mouse to a normal mouse, . . .
the normal mouse becomes obese
Low FODMAP diet
- increased actinobacteria
- high FODMAP diet decreased abundance of bacteria involved in gas consumption
- reduced IBS
Shown in human observational studies + human interventional studies
Fibre & prebiotics
- increased microbiota diversity & SCFA production
- reduced type 2 diabetes & cardiovascular disease
- human obs studies + human interventional studies
How is the microbiome distributed throughout the body of the baby?
newborn babies have similar microbiomes in all sections of their bodies
- vaginal delivery have vaginal-like microbiomes
- caesarian delivery have skin-like microbiomes
The developing gut microbiome - early gut colonization has 4 phases
- Phase 1: Sterile gut
- Phase 2: initial acquisition - vagina, feces, hospital
- Phase 3: breast-feeding or bottle-feeding (different)
- Phase 4: Start of solids; move to adult flora
(Bifidobacteria remain key flora into adutlhood)
What is the difference in microbiomes b/w breast-fed vs bottle-fed babies?
- Breast fed more bifidobacteria (up to 90% of flora)
- Bottle fed more diverse; more Bacteroides, and Clostridial species
What are HMOs good for?
- they help bifidobacteria to grow & proliferate
- it competitively excludes pathogens from binding
- decoy for other microbes to attach to so they can be removed
- macronutrient digestion
- development of mucus layer
- HMOs affect bifidobacteria and promote growth of SCFAs
What is the effect of SCFA?
- help the growth of enterocytes and use them for energy harvest
- also involved with anti-inflammatory cytokines
- involved in immune modulation
Adult microbiome
- both genetic and environmental factors
- increasing diversity of flora as awe age
- in some newer PCR studies, up to 92% of the flora in adults were “novel” species
- serial stool collections show remarkable stability by an individual
Who first discovered microorganisms in the body?
Leeuwenhoek
- made a simple microscope
- found them in dental plaque and a stool sample
What challenge did Robert Koch find during the “golden age of bacteriology”?
- He required a solid medium that would support bacterial growth (to obtain pure cultures)
- his associate’s wife eventually suggested agar
- he impact on medical microbiology was immediate
What was the great plate count anomaly?
most of the microscopically observable microorganisms in an environmental sample could not be cultured in the lab
Usually, between 1.0% and 0.1% of the total bacteria could be accounted for by the standard plating method.
What amazing discovery did Woese make?
- he used a molecular phylogenetic approach for tracing human history
- he couldn’t use fossils bc most microbes can’t be fossilized
- he took cellular
ribosomes and undertook a comparative analysis of the sequences of one component: the
small subunit ribosomal RNAs or SSU rRNAs. - Woese reasoned that the
similarities and differences between these sequences (i.e., the order of
the four chemical bases—adenine, uracil (or thymine in DNA), cytosine,
and guanine) would reflect the phylogenetic relationships of the organisms from which they were obtained.
Based on the SSU rRNAs, what did Woese find?
he found that there are 3 domains of life, 2 prokaryotic (bacteria, archae), and 1 eukaryotic
- eukaryotic cells have 18S rRNas
- prokaryotic cells have 16S rRNAs
The SSU rRNA sequences not only contained unique short sequences that defined the three domains but also contained unique sequences that permitted assignment of cells to specific phyla
Which 2 primary inventions expended the range of applications for SSu rRNA analysis?
- Sanger sequencing in 1977
- polymerase chain reaction (1980)
permitted the cloning and sequencing of SSU
rRNA genes from DNA samples extracted directly from environmental
samples
What is metagenomics?
sequencing of entire genomes
What happens in whole-genome shotgun sequencing?
DNA sequences are randomly broken up (shot-gunned) into smaller DNA fragments; computer programs reassemble the complete sequence by taking these fragments and looking for regions of overlap
What do metagenome sequences provide info about?
they provide info about which bacteria exist in a microbial population and at least a partial prediction of what functions are encoded by its genes
Proportion of microbes in / on the body
- 1% to 3% of body mass
- bacteria vastly outnumber the other kinds of microorganisms
- ratio of bacterial cells to human cells is about 1.3 to 1 (NOT 10 to 1)
What was the goal of the Human Microbiome Project?
to catalog the bacterial taxa associated with different body habitats on the healthy human
What were the main findings of the Human Microbiome Project?
- there is substantial variation in microbial community composition & diversity at diff body habitats
- oral & intestinal communities were very diverse, vaginal communities less so
- no taxa were universally present in all body
habitats and individuals - there is a high degree of stability in microbiome variety within an individual