Transcript Escherichia coli - Illinois Sustainable Technology Center

Ecological Impacts of Pharmaceutical
and Personal Care Products in Illinois
Rivers and Streams
John Kelly
Loyola University Chicago
Department of Biology
Emma Rosi-Marshall
Cary Institute for Ecosystem Studies
Teresa Chow / Monte Wilcoxon
Illinois Sustainable Technology Center
Pharmaceutical and Personal Care
Products (PPCPs)
• Prescription and non-prescription drugs:
•
•
•
•
•
•
Antibiotics
Analgesics
Antihistamines
Antacids
Hormones
Caffeine
• Household Products
• Antibacterials / Antimicrobials
Release of PPCPs to Environment
• Domestic wastewater
– PPCPs have been detected in
domestic wastewater
• Wastewater treatment plants
– Remove most PPCPs well
• Removal of PPCPs is generally
not 100%
• Low level PPCP release to
surface waters via effluent
– Some PPCPs partition to biosolids
• Land application of biosolids can
release PPCPs due to leaching
PPCPs have been detected in U.S.
rivers and streams
• Kolpin et al. 2002 (USGS)
– Suite of 95 contaminants
– Surveyed 139 streams in 30 states
• Biased toward streams susceptible
to contamination
• Analyzed water only
– Found 82 of the 95 contaminants
– Contaminants in 80% of streams
– Concentrations were generally low
• 95% of the concentrations
measured were below 1 mg/L
Ecological effects of most PPCPs
are unknown
• PPCPs are biologically active
• PPCPs are likely to have
ecological effects:
• Classes of organisms
• Acute vs. Chronic effects
• Community composition
• Ecosystem processes
Triclosan
•
•
•
•
5-chloro-2-(2,4-dichlorophenoxy) phenol
Potent antibacterial compound
Discovered in 1960s
Mode of action
– Blocks synthesis of lipids in bacteria
– Inhibits enoyl-acyl carrier protein reductase
• Used primarily in hospitals as an antiseptic and
disinfectant
Triclosan
• Between 1992 and 1999 700 products containing
triclosan entered the consumer market in the US
• Currently found in:
– Soap
– Detergents
– Cleaning Products
– Toothpaste / Mouthwash
– Paint
• Has been embedded in
– Plastics
– Textiles
Triclosan has been detected in U.S.
rivers and streams
• Kolpin et al. 2002 (USGS)
– The most frequently detected compounds
were:
• Diethyltoluamide (insect repellant)
• Caffeine
• Triclosan (58% of streams)
• Trichloroethylphosphate (fire retardant)
– Concentrations in water were generally low
• Triclosan high 2.3 ug/L
• Triclosan median 0.14 ug/L
Why should we be concerned about
triclosan in streams ?
Triclosan
• Low solubility in water, lipophilic
• Has been detected in lake and estuarine sediments
(Singer et al. 2002, Miller et al. 2008).
• Has been shown to persist in the environment,
especially under anaerobic conditions (Ying et al. 2007)
• Broad spectrum antibacterial compound
Bacteria are significant contributors to
stream ecosystems
• Organic matter decomposition
– Nutrient release
• Nutrient Cycling
– Nitrogen
• Biomass production
– Base of stream food webs
• Denitrification
– Ameliorates nitrogen pollution
Bacteria can develop
triclosan resistance
• Growth of triclosan sensitive
E. coli for 200 generations in
presence of low levels of
triclosan produced triclosan
resistant mutants (George
and Levy, 1983)
• Molecular analysis of mutants
revealed mutation in gene
fabI (which encodes enoyl
acyl carrier protein
reductase) (McMurry,
Oethinger, and Levy 1998)
Escherichia coli
Possible links between triclosan
resistance and antibiotic resistance
• Triclosan exposure has been
shown to increase resistance to
antibiotics in
– Pseudomonas aeruginosa
• (Chuanchuen et al., 2001)
– Escherichia coli
• (Braoudaki and Hilton, 2004).
Pseudomonas
aeruginosa
Questions to be addressed by our project
• Are there detectable levels of
triclosan in waters and
sediments of Illinois rivers and
streams?
• Are the levels of triclosan in
Illinois rivers and streams
impacting the resident bacterial
communities?
• How does triclosan impact the
composition and function of
sediment bacterial
communities?
Experimental Design
Field Survey
Artificial Streams
Artificial Streams: Pilot Study
• Two streams received:
• Sand and pea gravel (4:1)
• 200g of shredded leaves
• 100g sediment from
Nippersink Creek
• Monitored weekly:
• Bacterial community size
• Heterotrophic plate counts
• Triclosan resistance
• Growth on triclosan
amended plates
• Triclosan added week 8 (16ug/L)
Artificial Streams: Plate Counts
Triclosan Addition
Bacterial Cells / G Dry Sediment
450000
Stream 1
400000
Stream 2
350000
300000
250000
200000
150000
100000
50000
0
3
4
5
6
7
Week
8
9
10
11
Artificial Streams: Triclosan Resistance
Triclosan Addition
4.00%
Stream 1
Percent Triclosan Resistance
3.50%
Stream 2
3.00%
2.50%
2.00%
1.50%
1.00%
0.50%
0.00%
8
9
10
Week
11
Artificial Streams: Next Steps
• Replicated experiment
– 5 streams per treatment
• Three treatments
– No triclosan
– Low triclosan
– High triclosan
• We will monitor
– Bacterial community size
– Triclosan resistance
– Activity
• Respiration, denitrification
– Community composition
Field Survey
• Sites sampled to date
• Assay in progress
– Non-impacted site
– Microbial activity
• Nippersink Creek, McHenry
– Respiration
County IL
– Denitrification
– Heavily Impacted: Urban WWTP
– Community
• North Shore Channel
composition
– Moderately Impacted: Suburban
– Triclosan
WWTP
concentration
• West Branch Dupage River
• Assays completed
– Bacterial community size
– Triclosan resistance
Field Survey: Nippersink Creek
• Total cell counts 3.7 x 105
cfu / g dry sediment
• Triclosan resistance 0.66%
Field Survey: North Shore Channel
• Receives effluent from the North
Side WWTP
• Serves over 1.3 million people
• Residing in a 141 square mile
area including the City of
Chicago north of Fullerton
Avenue and the northern
Cook County suburbs.
• The Plant has a design
capacity of 333 million gallons
per day
Field Survey: North Shore Channel
Triclosan Resistance
200,000
3.5%
175,000
3.0%
Triclosan Resistance
CFU g-1 dry sediment
Bacterial Counts
150,000
125,000
100,000
75,000
50,000
25,000
2.5%
2.0%
1.5%
1.0%
0.5%
0.0%
0
Upstream
Downstream
Upstream
Downstream
Field Survey: West Branch DuPage River
• Receives effluent from the West
Chicago WWTP
• The Plant has a design capacity
of 20 million gallons per day
Field Survey: West Branch DuPage River
Bacterial Counts
Triclosan Resistance
Triclosan Resistance
CFU g-1 dry sediment
250,000
200,000
150,000
100,000
50,000
0
Upstream
Downstream
2.0%
1.8%
1.6%
1.4%
1.2%
1.0%
0.8%
0.6%
0.4%
0.2%
0.0%
Upstream
Downstream
Field Survey
• Sites sampled to date
• Assay in progress
– Non-impacted site
– Microbial activity
• Nippersink Creek, McHenry
– Respiration
County IL
– Denitrification
– Heavily Impacted: Urban WWTP
– Community
• North Shore Channel
composition
– Moderately Impacted: Suburban
– Triclosan
WWTP
concentration
• West Branch Dupage River
• Assays completed
– Bacterial community size
– Triclosan resistance
Field Survey: Next Steps
• Additional Field Sites
– Moderately Impacted: Suburban WWTP
– East Branch DuPage River
– Agricultural Sites
– Sites receiving biosolids
amendments
Acknowledgements
• Collaborators
• Emma Rosi-Marshall
• Teresa Chow
• Monte Wilcoxon
• Students
• Brad Drury
• Diana Deavila
• Funding
• Illinois Sustainable Technology Center