Lesson 11 Slide

Environmental Biotechnology

🌍 1. What Are Micropollutants?

Micropollutants are organic or inorganic substances present in trace amounts (µg/L to ng/L) in water. They include 💊 pharmaceuticals, 💅 personal care products, 🌾 pesticides, and ⚙️ industrial chemicals. Even at low concentrations, they can disrupt ecosystems and harm organisms.

💡 Why they matter:

Modern wastewater treatment plants (WWTPs) aren’t designed for these trace pollutants. The goal of future treatment is higher effluent water quality and stable operation, with resource recovery (energy, nutrients, materials).


🚰 2. Sludge and Wastewater Overview

Each year, Denmark produces about 365,000 tonnes of surplus sludge (5–30% dry matter). It’s used in:

  • Agriculture (40–60%) 🌾
  • Biogas (10–20%) 🔋
  • Thermal treatment (5–15%) 🔥
  • Export & recycling (<5%)

Sludge contains energy (15–30 MJ/kg dry matter) and valuable resources, but also micropollutants — making treatment complex.


💊 3. Pharmaceuticals in WWTPs

Studies (Falås et al., 2012) show that pharmaceuticals enter and leave WWTPs largely intact — conventional processes remove only part of them. Each plant varies in efficiency depending on design, operation, and hydraulic retention time.

Micropollutants often combine in cocktails, causing additive, antagonistic, or synergistic effects on organisms. ☠️


🔄 4. How Can We Remove Micropollutants?

Main routes of removal:

  1. Evaporation
  2. Photooxidation
  3. Adsorption
  4. Chemical oxidation
  5. Biodegradation (most sustainable 💚)

Typical treatment technologies:

  • Physical: Membrane processes (nanofiltration, reverse osmosis)
  • Biological: Transformation/degradation by microbes
  • Oxidation: Ozone treatment
  • Adsorption: Activated carbon
  • Combinations: e.g., Ozone + GAC (Granular Activated Carbon)

⚗️ 5. Example – Triclosan (Antibacterial agent)

A mass balance shows:

  • 5% remains in effluent
  • 30% bound to sludge
  • 10% forms bound residues

  • 10% becomes methylated derivative
  • 40% is unknown

🧩 Bound residues = chemicals attached to solids (plastics, fibers, etc.), often hidden from standard tests.


⚖️ 6. Pros & Cons of Treatment Methods

MethodAdvantagesDisadvantages
Activated CarbonTreats many MPs; low costPoor removal of some (BAM/DMS); maintenance; disinfection needed
Chemical Oxidation (Ozone)Effective on many MPsToxic byproducts; costly
Filtration (NF/RO)High efficiency; no byproductsVery expensive; high maintenance; water loss; MP upconcentration

🏭 7. Factors Influencing Removal

Different WWTPs (Aalborg East, West, Aabybro, Hirtshals) show varying rates due to:

  1. Pollutant loading
  2. Retention time
  3. Plant design
  4. Operational control

🔬 8. Studying Degradation Methods

When pathways are unknown, scientists combine molecular and ecological tools:

  • FISH & Amplicon sequencing → identify and quantify bacteria.
  • MAR-FISH → tracks radioactive substrate uptake at the single-cell level.
  • SIP (Stable Isotope Probing) → identifies degraders and resistant species via ^13C labeling.

Example: Only 0.1‰ of biomass degraded EE2 (a hormone pollutant). Tiny but significant!


🧫 9. Measuring Activity

Biometer flasks detect degradation by measuring radioactive CO₂ production. GC-MS/MS and toxicology assays confirm compound disappearance and transformation. Some pollutants (like Prozac) can alter animal behavior — e.g., make fish aggressive! 🐟💢


🧱 10. Biofilm vs. Suspended Biomass

Adding biofilm carriers (MBBR systems) improves removal of slowly degradable compounds 8–10× compared to conventional activated sludge. Biofilms provide:

  • More surfaces 🧩
  • Higher bacterial retention
  • Longer residence time

🌿 11. Bioaugmentation & Biostimulation

Introducing specialized microbes or nutrients can enhance degradation. However, exogenous strains often fail due to:

  • Competition from native microbes 🦠
  • Predation and stress
  • Delivery challenges

Effectiveness is usually short-lived. Still, it’s useful for targeted removal or resilience boosting.


🧬 12. Genomics & Metagenomics

Workflow:

  1. DNA extraction
  2. Sequencing (reads → contigs → assembly)
  3. Classification:
    • Who is there? (Phylogenetic)
    • What can they do? (Functional)

Metagenome-Assembled Genomes (MAGs) reconstruct genomes from complex communities — sometimes representing <0.1% of the total population.


🧠 13. From Genes to Function

Challenges:

  • Gene annotation bias
  • Multiple protein functions
  • No direct link to activity
  • High microdiversity
  • “Transitive catastrophe”: one wrong annotation propagates across databases

👉 COMBREX helps bridge computational predictions with experimental validation.


🧩 14. Pathway Discovery & “Omics” Integration

Genome mining can identify degradation-related genes, but expression (epigenetics) matters too. Tools like EAWAG-BBD Pathway Prediction System (PPS) simulate microbial degradation routes based on known reactions.

By combining:

  • Metabolomics 🧪
  • Genomics 🧬
  • Proteomics 🧫

→ Researchers can reveal complete degradation pathways.


🧪 15. Example: Gemfibrozil Degradation

Gemfibrozil (a lipid-lowering drug) undergoes:

  • “Unspecific” degradation by Cytochrome P450 enzymes
  • Upregulation of transport proteins (TonB, OmpW) and detox systems
  • Iron-mediated dechlorination and antioxidant defense (via Glutathione reductase)

Each response helps the cell survive and transform the compound.


💥 16. Example: EE2 and Ibuprofen

Exposure to EE2 (synthetic estrogen) changes expression of many molecular functions (membrane transport, oxidoreductases, etc.). Ibuprofen degradation involves enzymes:

  • AMP-dependent synthases
  • Dioxygenases (Rieske Fe-S domain)
  • Enoyl-CoA hydratases

🧬 Together, they break down ibuprofen through aromatic ring cleavage and oxidation.


🧫 17. Applying Genomic Knowledge

By identifying marker genes and pathways, we can:

  • Predict microbial community potential 🧮
  • Engineer bioreactors for enhanced xenobiotic degradation
  • Design stable, efficient microbial ecosystems for wastewater treatment

Goal: Move from describepredictcontrol microbial behavior.


⚙️ 18. Final Concept – Microbial Ecosystem Engineering

Steps for sustainable bioremediation:

  1. Describe the community
  2. Understand its function
  3. Explain how species interact
  4. Predict their roles
  5. Control the system to maintain performance

→ Outcome: a bioreactor with predictable, stable, and effective pollutant degradation.

Quiz

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