continuous improvement and evolution of michigan’s water ......continuous improvement and...
TRANSCRIPT
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Continuous Improvement and Evolution of Michigan’s
Water Quality Testing
Presented By Alexandra Strohm
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Outline
•Background
•History of Water Quality Testing
•Current Testing Methods
•Applications
•Future Methods
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Background
Alexandra Strohm
• Junior Michigan State University
• Biochemistry and Molecular Biology, Pharmacology and Toxicology
• MDEQ Student Assistant Water Toxic Unit
• Lyman Briggs Molecular Biology PCR Lab MSU
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“Short History of Water Pollution Abatement in Michigan”
• Francis Frost wrote a short a summary of Michigan’s history of water pollution
• Frost was one of 3 member Michigan Water Resources Commission staff
• Documented the struggle of making Michigan’s water quality a priority
1923 1965 1968 1929 1935 1948 1952 1972 1990 2000 1997 1988 1995
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“Short History of Water Pollution Abatement in Michigan”
• From 1923 to 1929 pollution control personnel were divided between several divisions with separate budgets Problem
• Act 245 Public Acts of 1929 created The Stream Control Commission Act • Towns that were ordered to build sewage plants tried to abolish commission
until about 1946
1923 1965 1968 1929 1935 1948 1952 1972 1990 2000 1997 1988 1995
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“Short History of Water Pollution Abatement in Michigan”
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1923 1965 1968 1929 1935 1948 1952 1972 1990 2000 1997 1988 1995
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1972 Clean Water Act
• NPDES (National Pollutant Discharge Elimination System) demanded permits for discharge and waste
• Limits were set to meet standards to make water fishable and swimmable
• Grants awarded for wastewater treatment
1923 1965 1968 1929 1935 1948 1952 1972 1990 2000 1997 1988 1995
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MDEQ BEACH Act
• 1995 MDEQ was created by Governor John Engler
• In 2000, BEACH (Beaches Environmental Assessment and Coastal Health) Act passed
• EPA provides grants for beach monitoring and notification programs
1923 1965 1968 1929 1935 1948 1952 1972 1990 2000 1997 1988 1995
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Water Quality Standards
Michigan Public Health Code and Rule 323.1062(1) of the Part 4. Water Quality Standards (Promulgated pursuant to Part 31 of the Natural
Resources and Environmental Protection Act, 1997 PA 451, as amended)
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MDEQ E. coli Testing
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Idexx Colilert-18 Quanti Tray Method
• MPN determines E. coli density
• ‘Defined Substrate Technology’ (MUG and ONPG nutrient indicators)
• Synthesizes fluorescent E. Coli
• ONPG is substrate for B-galactosidase (enzyme) which turns the coliform yellow
• MUG is substrate for B-glucuronidase (enzyme) which causes E. coli to fluoresce
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What is PCR?
Polymerase Chain Reaction
Amplify target segments of DNA Billions of copies!!!
Test amount or presence/absence of specific DNA markers
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PCR: DNA Synthesis in Vitro
Cocktail ingredients:
Template DNA
Oligonucleotide Primers
dNTPs
Taq Polymerase
PCR Buffer
Mg2+
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What is PCR?
• Denaturation (95 degrees Celsius)
• Annealing (~68 degrees Celsius)
• Elongation (72 egress Celsius)
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The Problem
“In the United States alone, it is estimated that 39.2% of all rivers, lakes, and streams are unsafe for recreational use, with fecal pathogens as the number one cause of impairment”
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How is qPCR useful in Water Testing?
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The Problem
“The results showed…Enterococcus measured by QPCR can predict GI illness after swimming in fecally contaminated fresh water…Incorporation of rapid measurements such as these into a regulatory framework has the potential to improve beach management decisions and protect swimmers’ health.”
“Ultimately, the use of faster indicators of recreational water quality will result in the ability to make decisions about recreational water quality on the day of sample collection. This, in turn, could lower GI illnesses in communities, especially in those dependent on beach-related tourism.”
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1
10
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1000
10000
Singing Bridge
MPN_Singing GEC_Singing
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y = 0.1505x + 20.386 R² = 0.1283
1
10
100
1000
10000
1 10 100 1000 10000
IDEX
X
EPA Method
Holland
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y = 0.4688x + 30.961 R² = 0.5267
1
10
100
1000
10000
1 10 100 1000 10000
IDEX
X
EPA Method
Singing Bridge
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Source Tracking: How do fix the problem?
• Identify the sources of E. coli present in a water sample
•Genetic markers in organisms
•Potential path to successful remediation
•qPCR
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Chrysler
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0
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2009 2010 2011 2012 2013 2014 2015 2016
E.co
li C
olo
ny
Form
ing
Un
its
pe
r 1
00
mL
Wat
er
Year
Chrysler Park: Individual Sample E.coli Concentrations (2009-2016)
Individual Sample of E.coliConcentration
TBC Maximum
Linear (Individual Sample ofE.coli Concentration)
Linear (TBC Maximum)
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0
200
400
600
800
1000
1200
1400
2009 2010 2011 2012 2013 2014 2015 2016
E. c
oli
CFU
in p
er
10
0 m
L W
ate
r
Year
Chrysler Park: Individual Sample 50th and 95th Percentile E. coli Concentration (2009-2016)
50th Percentile
95th Percentile
Linear (50th Percentile)
Linear (95th Percentile)
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Brimley State Park
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Brimley State Park
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0
500
1000
1500
2000
2500
2010 2011 2012 2013 2014 2015 2016
E. c
oli
Co
nce
ntr
atio
n in
MP
N/1
00
mL
Wat
er
Year
Brimley State Park: Individual Sample E. coli Concentrations (2010-2016)
Individual Sample E. coliConcentrations
TBC WQS
Linear (Individual Sample E. coliConcentrations)
Linear (TBC WQS)
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0
500
1000
1500
2000
2500
2010 2011 2012 2013 2014 2016
E. c
oli
Co
nce
ntr
atio
n in
MP
N/1
00
mL
Wat
er
Year
Brimley State Park: Individual Sample 50th and 95th Percentile E. coli Concentrations (2010-2016)
50th Percentile
95th Percentile
Linear (50th Percentile)
Linear (95th Percentile)
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Challenges with Source Tracking
•Fails near LLOQ
•Genetic markers must be highly specific
•Primers could bind to multiple organism DNA (Dogs and Human cross)
•Best markers and criteria still being developed
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Source tracking: Human Vs. Dogs • “Study estimates that 39.1 percent of human pathogens also infect
domestic animals”
• Used genome fragment enrichment (GFE) to identify nonribosomal microbial genetic markers for PCR-based testing of dog fecal contamination
• DG3, DG37, DG72 markers
• 244 Fecal samples below LOD for noncanines (humans)
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Digital PCR • Eliminates the need for a standard curve
• Plasson statistics/ corrections
• More sensitive smaller differences detected
• Less reagents needed (no standards, less wells used)
• No cross reactions
• Partitions sample into droplets or chambers
• Flow cytometer checks droplets
• Measures positive or negative/ accepted or not accepted
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Thank You!