What’s living in your water?™
DNA-Based
Microbial Analysis
Microbe Detectives
Trevor Ghylin, P. E.
Global Water Center, Milwaukee, WI
trevor.ghylin@microbedetectives.com
414-217-7784
www.microbedetectives.com
Company Introduction
• My Background
o
o
o
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Professional Engineer – CH2M Hill
WEF Canham Graduate Studies Fellowship
NIH Biotechnology Training Program Fellow
PhD Candidate – Biotechnology
• Founded in 2012 to improve access to DNA sequencing
services
o Global Freshwater Seed Accelerator Winner
o Rising Star Award - Early Stage Symposium
• Advisory Board - Current and former IWA presidents
• Key Developers - Biomolecular Engineer, Bioinformaticist
• Clients – CH2M Hill, Milwaukee Water, Consulting Firms
Current paradigm
Problem: >99% of microbes can’t be cultured or
identified under a microscope
Solution
DNA-Based Microbial
Analysis
Collect
Sterilize, Preserve, Ship
Environmental
Sample (50mL sterile
bottle)
Add ethanol to 70% concentration to sterilize.
Dilute with water to 25% for preservation and
shipping (Shipping via Fedex: 5
days/$85USD/51GBP
Concentrate
Centrifuge or Filtration
Extract DNA/Purify
DNA Sequencing
PCR Amplification/DNA Sequencing
ATGCATT…...
Assign Taxonomy (Proprietary) Nitrosomonas
Bioinformatics/DNA Processing/Taxonomic Assignment
Interpretation of Results
(Proprietary)
Healthy AOB population
Example DNA Data
>594682
TACGGAGGATCCAAGCGTTATCCGGATTTATTGGGTTTAAAGGGTACGTAGG
CGGACCCG
TCAGTCAGTGGTGAAAGTTTGCAGCTTAACTGTAAAATTGCCATTGAAACT
ACGGGTCTT
GAGTGTAAATGAGGTAGGCGGAATGTGTTGTGTAGCGGTGAAATGCTTAGA
TATAACACA
GAACACCAATTGCGAAGGCAGCTTACTGGGATACAACTGACGCTGAGGCA
CGAAAGCGTG
GGGATCAAACAGG
>594511
TACGGAAGGTCCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAACGCAG
GCTGGAGAT
TAAGCGTGCTGTGAAATGTACCGGCTCAACCGGTGACGTGCAGCGCGAAC
TGGTTTCCTT
GAGTGAGTACGACGTCAGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGA
TATCACGAA
Database of Wastewater Microbes (Proprietary)
Solution Cont’d
• Benefits
Solution Cont’d
o Wastewater Treatment
• Solve treatment problems
• Reduce energy and chemical consumption
• Maximise digester energy production
• Maximise effluent quality
o Membrane Treatment
• Solve Biofouling issues
• Reduce chemical consumption
• Reduce operational and maintenance labor
• Increase membrane life
o Drinking Water
• Solve taste/odor issues
• Solve coliform issues
• Ensure quality and safety
• Identify and fix distribution system issues
Case Studies
Membrane Biofouling
Membrane Biofouling Cont’d
Wastewater Treatment
• Municipal SBR – Filamentous Bulking in
Spring
• SVI~300
• Poor settling, poor effluent quality
• Potential to exceed permit TSS limit
Microscopy Results
Drinking Water Distribution
System Case Study
Costs
• Project specific
• Membrane Biofouling
o $2,000USD (1,200 GBP) analysis (save >$10,000
USD/6,000GBP in chemical/operational)
• Wastewater Treatment
o $600USD (350 GBP) for 1 sample ($1000USD/600GBP) with
microscopy)
• Drinking Water
o Provide distribution system insurance for pennies per resident
• Business Model
o Mail-in service business with additional consulting fee
($140/hr USD/85 GBP)
o Return on investment can be extremely high
Status and Path Forward
• Currently serving clients in drinking water and
wastewater
• Pilot studies
o Wastewater Treatment
• Year-long pilot; weekly or monthly samples
o Drinking Water Distribution System
• Single sampling event; collect samples from various
locations in the distribution system
• Multiple cities to generate more data
o Membrane Biofouling
Status and Path Forward Cont’d
• Plans
o Pilot studies
o Continue to serve clients and build our knowledge,
refine our databases
o Milwaukee distribution system project
• Support Needed
o Wastewater Treatment Demonstration
o Membrane Biofouling Demonstration
o Drinking Water Distribution System Demonstration
Summary and next steps
• DNA analysis is economical
o Superior to existing petri dish and microscopy methods
• Need partners to do more extensive pilot testing work
and demonstrate results
o Reduced energy and chemical consumption
o Improved drinking water quality
o Improved membrane performance
Thank You
What’s living in your water?
Global Water Center, Milwaukee, WI
trevor.ghylin@microbedetectives.com
414-217-7784
www.microbedetectives.com
LinkedIn: Trevor Ghylin