22  Metagenomics and the Microbiome

22.1 Studying Communities of Microbes

22.1.1 What Is Metagenomics?

Metagenomics = Studying genetic material from environmental samples containing MANY different species

Meta = beyond or among Genomics = studying genomes

So: Studying the genomes of entire communities at once!

Think of it like:

  • Analyzing a whole forest instead of one tree

  • Studying a city instead of one person

  • Reading all books in a library at once

22.1.2 The Microbiome

Microbiome = All the microorganisms living in a particular environment

Examples:

  • Human gut microbiome: Trillions of bacteria in your intestines

  • Soil microbiome: Microbes in dirt

  • Ocean microbiome: Microbes in seawater

  • Skin microbiome: Bacteria on your skin

Key fact: There are more bacterial cells in/on you than human cells!

  • ~30 trillion human cells

  • ~40 trillion bacterial cells

  • You’re more microbe than human (by cell count)!

22.2 Why Study Microbiomes?

22.2.1 They’re Everywhere and Important!

In your body:

  • Help digest food

  • Make vitamins (K, B12)

  • Train immune system

  • Protect from pathogens

  • Affect mood and behavior!

In the environment:

  • Decompose dead material

  • Fix nitrogen (make fertilizer)

  • Clean up pollution

  • Cycle nutrients

  • Produce oxygen (ocean microbes!)

Without microbes: Life on Earth would collapse!

22.3 The Challenge: Unculturable Microbes

22.3.1 Most Microbes Can’t Be Grown in Lab

The problem:

  • Traditional microbiology: Grow bacteria on plates

  • But 99% of environmental microbes won’t grow in lab!

  • Called “unculturable”

Why they won’t grow:

  • Need specific conditions

  • Grow too slowly

  • Depend on other microbes

  • Unknown nutritional needs

Result: We were missing most microbial diversity!

22.3.2 The Solution: Culture-Independent Methods

Metagenomics breakthrough:

  • Don’t need to grow them!

  • Extract DNA directly from environment

  • Sequence all DNA together

  • Identify all microbes present

It’s like:

  • Instead of inviting each person to interview

  • Reading everyone’s ID cards at once

  • Getting census data without asking questions

22.4 How Metagenomics Works

22.4.1 The Workflow

1. Sample Collection

  • Collect environmental sample

  • Soil, water, feces, skin swab, etc.

  • Contains billions of microbes

2. DNA Extraction

  • Break open all cells

  • Extract all DNA

  • Mix of DNA from thousands of species!

3. Sequencing

  • Sequence all DNA fragments

  • Millions of reads

  • Don’t know which species each fragment comes from (yet!)

4. Analysis

  • Identify which microbes are present

  • How many of each?

  • What genes do they have?

  • What can they do?

22.4.2 Two Main Approaches

16S rRNA Sequencing (targeted):

  • Sequence only 16S rRNA gene

  • Present in all bacteria

  • Like a bacterial barcode

  • Tells you WHO is there

  • Cheap and fast!

Shotgun Metagenomics (whole genome):

  • Sequence ALL DNA

  • More expensive

  • Tells you WHO is there AND what they can do

  • Functional information

22.5 The Human Microbiome

22.5.1 We’re Walking Ecosystems!

Where microbes live on/in you:

  • Gut (most abundant): 100 trillion bacteria

  • Skin: Different microbes on different body parts

  • Mouth: Hundreds of species

  • Nose: Unique communities

  • Vagina: Dominated by Lactobacillus (usually)

It’s personal: Everyone has unique microbiome!

  • Like a fingerprint

  • Affected by diet, genetics, environment, medications

22.5.2 The Gut Microbiome

Most studied and important

Major bacterial groups (phyla):

  • Firmicutes (40-60%)

  • Bacteroidetes (20-40%)

  • Actinobacteria (smaller %)

  • Proteobacteria (smaller %)

  • Plus archaea, fungi, viruses!

What they do:

1. Digest Food:

  • Break down complex carbohydrates we can’t digest

  • Fiber, resistant starch

  • Produce short-chain fatty acids (healthy!)

2. Make Vitamins:

  • Vitamin K (blood clotting)

  • B vitamins (B12, biotin, folate)

  • We couldn’t make these without microbes!

3. Train Immune System:

  • Teach immune cells friend vs. foe

  • Without microbes: Allergies, autoimmune diseases

  • “Old friends” hypothesis

4. Protect from Pathogens:

  • Occupy space (no room for bad guys)

  • Produce antimicrobial compounds

  • Compete for resources

5. Produce Neurotransmitters:

  • Serotonin (mood)

  • GABA (calming)

  • Dopamine

  • “Gut-brain axis”!

22.5.3 Dysbiosis: When Microbiome Goes Wrong

Dysbiosis = Imbalanced microbiome

Associated with:

  • Obesity

  • Type 2 diabetes

  • Inflammatory bowel disease (IBD)

  • Allergies

  • Autism (controversial)

  • Depression and anxiety

  • Autoimmune diseases

Causes of dysbiosis:

  • Antibiotics (kill good bacteria too!)

  • Poor diet (low fiber)

  • Stress

  • Lack of sleep

  • C-section birth (miss mom’s microbes)

  • Formula feeding (vs. breastfeeding)

22.6 Environmental Metagenomics

22.6.1 Soil Microbiome

Incredibly diverse:

  • Teaspoon of soil: 1 billion bacteria

  • Thousands of different species

  • More diversity than gut!

What they do:

  • Decompose organic matter

  • Cycle nutrients (nitrogen, phosphorus)

  • Help plants grow

  • Carbon storage (climate change!)

Applications:

  • Agriculture (healthy soil = healthy crops)

  • Bioremediation (clean up pollution)

  • Carbon sequestration

22.6.2 Ocean Microbiome

Craig Venter’s Global Ocean Sampling:

  • Sailed around world

  • Sampled ocean microbes

  • Discovered MILLIONS of new genes!

Cyanobacteria (like Prochlorococcus):

  • Most abundant photosynthesizer on Earth!

  • Produce 20% of Earth’s oxygen

  • Tiny but critically important

Marine microbes:

  • Base of food chain

  • Carbon cycling

  • Climate regulation

  • Chemical diversity (drug discovery!)

22.7 Metagenomics Applications

22.7.1 Medicine

Fecal Microbiota Transplant (FMT):

  • Transfer healthy microbiome to patient

  • Cures C. difficile infection (90% success!)

  • Being tested for other diseases

Personalized Medicine:

  • Microbiome affects drug metabolism

  • Could predict treatment response

  • Tailor treatments to individual microbiome

Diagnostics:

  • Microbiome signatures for diseases

  • Early detection

  • Non-invasive testing

22.7.2 Agriculture

Improve Crop Yield:

  • Select beneficial soil microbes

  • Reduce fertilizer needs

  • Increase drought tolerance

  • Probiotics for plants!

Animal Health:

  • Livestock microbiome optimization

  • Reduce antibiotics use

  • Improve nutrition

22.7.3 Biotechnology

Bioprospecting:

  • Find new enzymes from microbes

  • Industrial applications

  • Laundry detergents

  • Biofuels

Waste Treatment:

  • Sewage processing

  • Decompose pollutants

  • Generate biogas

22.7.4 Environmental

Bioremediation:

  • Clean up oil spills

  • Remove heavy metals

  • Degrade plastics (potentially!)

Climate Change:

  • Understanding carbon cycle

  • Methane production/consumption

  • Ocean acidification impacts

22.8 The Human Microbiome Project

22.8.1 Mapping Our Microbial Partners

HMP (2007-2016):

  • NIH-funded project

  • Characterize microbiome of healthy humans

  • 300 healthy people

  • Multiple body sites

Key findings:

  • Huge diversity (10,000+ species)

  • Everyone has unique microbiome

  • Core functions shared (even if species different)

  • Variation is normal

Impact:

  • Reference dataset

  • Enables disease studies

  • Launched microbiome research boom

22.9 Microbiome Manipulation

22.9.1 Can We Change Our Microbiome?

Methods:

1. Diet:

  • Fiber feeds good bacteria

  • Fermented foods (yogurt, kimchi, sauerkraut)

  • Prebiotics (food for probiotics)

  • Mediterranean diet linked to healthy microbiome

2. Probiotics:

  • Live beneficial bacteria

  • Yogurt, supplements

  • Evidence mixed (not all are helpful!)

  • Strain-specific effects

3. Antibiotics (when necessary):

  • Kill harmful bacteria

  • But also good ones

  • Use wisely!

4. Fecal Microbiota Transplant:

  • Extreme intervention

  • Very effective for some conditions

  • Being researched for other diseases

5. Exercise:

  • Associated with healthier microbiome

  • Increases diversity

  • Mechanisms unclear

22.10 Challenges in Metagenomics

22.10.1 Technical Difficulties

1. Complexity:

  • Thousands of species

  • Complex interactions

  • Hard to figure out who’s doing what

2. Computational:

  • Massive datasets

  • Need powerful computers

  • Analysis challenging

3. Causation vs. Correlation:

  • Microbiome difference ≠ cause of disease

  • Could be consequence

  • Hard to prove causation

4. Individual Variation:

  • Everyone’s different

  • Hard to define “healthy” microbiome

  • Personalized approaches needed

5. Functional Redundancy:

  • Different microbes can do same job

  • Focus on function, not just species

22.11 Virome: The Viral Microbiome

22.11.1 Viruses Everywhere!

Bacteriophages (viruses that infect bacteria):

  • 10x more abundant than bacteria!

  • Control bacterial populations

  • Transfer genes between bacteria

  • Largely unknown

Human viruses:

  • Many present without causing disease

  • Part of normal microbiome

  • Potential roles in health/disease

Phage therapy:

  • Use viruses to kill harmful bacteria

  • Alternative to antibiotics

  • Old idea, new interest!

22.12 Mycobiome and Beyond

22.12.1 Not Just Bacteria!

Mycobiome (fungi):

  • Present in gut, skin, etc.

  • Less studied than bacteria

  • Candida and others

  • Role in health unclear

Archaeome (archaea):

  • Methane-producing archaea in gut

  • Less abundant than bacteria

  • Unique metabolism

Parasites and protists:

  • Also part of microbiome

  • Complex interactions

  • Often pathogenic but not always

22.13 The Future of Microbiome Research

22.13.1 What’s Coming

1. Precision Microbiome Medicine:

  • Analyze your microbiome

  • Personalized probiotic prescriptions

  • Targeted microbiome manipulation

2. Microbiome Editing:

  • CRISPR for microbiome

  • Edit bacterial genes in situ

  • Remove harmful functions

3. Synthetic Microbial Communities:

  • Design optimal microbiome

  • Engineer bacteria for specific functions

  • Living therapeutics

4. Biomarkers:

  • Microbiome signatures for diseases

  • Diagnostic tests

  • Track treatment response

5. Environmental Applications:

  • Climate change mitigation

  • Pollution cleanup

  • Sustainable agriculture

22.14 Key Takeaways

  • Metagenomics studies genetic material from mixed communities

  • Microbiome = all microorganisms in an environment

  • More bacterial cells than human cells in your body!

  • Most microbes can’t be cultured (99%)

  • Two approaches: 16S rRNA (who’s there) vs. shotgun (what they do)

  • Gut microbiome: Digests food, makes vitamins, trains immunity, affects mood

  • Dysbiosis = imbalanced microbiome, linked to diseases

  • Human Microbiome Project mapped our microbial partners

  • Diet, probiotics, FMT can modify microbiome

  • Applications: Medicine, agriculture, environmental cleanup

  • Everyone has unique microbiome

  • Microbiome research is exploding field with huge potential


Sources: Information adapted from Human Microbiome Project, microbiome research literature, and metagenomics studies.