ecotoxicology of antimicrobial ppcps in il rivers and streams · •low level ppcp release to...
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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
• 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
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
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
• 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)
Bacteria can develop
triclosan resistance
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
• 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?
Questions to be addressed by our project
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)
0
50000
100000
150000
200000
250000
300000
350000
400000
450000
3 4 5 6 7 8 9 10 11
Ba
cte
ria
l C
ells
/ G
Dry
Se
dim
en
t
Week
Stream 1
Stream 2
Triclosan Addition
Artificial Streams: Plate Counts
0.00%
0.50%
1.00%
1.50%
2.00%
2.50%
3.00%
3.50%
4.00%
8 9 10 11
Pe
rce
nt T
ric
los
an
Re
sis
tan
ce
Week
Stream 1
Stream 2
Artificial Streams: Triclosan Resistance
Triclosan Addition
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
– Non-impacted site
• Nippersink Creek, McHenry
County IL
– Heavily Impacted: Urban WWTP
• North Shore Channel
– Moderately Impacted: Suburban
WWTP
• West Branch Dupage River
• Assays completed
– Bacterial community size
– Triclosan resistance
• Assay in progress
– Microbial activity
– Respiration
– Denitrification
– Community
composition
– Triclosan
concentration
• Total cell counts 3.7 x 105
cfu / g dry sediment
• Triclosan resistance 0.66%
Field Survey: Nippersink Creek
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
0
25,000
50,000
75,000
100,000
125,000
150,000
175,000
200,000
Upstream Downstream
CF
U g
-1d
ry s
ed
ime
nt
0.0%
0.5%
1.0%
1.5%
2.0%
2.5%
3.0%
3.5%
Upstream DownstreamT
ric
los
an
Re
sis
tan
ce
Bacterial Counts Triclosan Resistance
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
0
50,000
100,000
150,000
200,000
250,000
Upstream Downstream
CF
U g
-1d
ry s
ed
ime
nt
0.0%
0.2%
0.4%
0.6%
0.8%
1.0%
1.2%
1.4%
1.6%
1.8%
2.0%
Upstream DownstreamT
ric
los
an
Re
sis
tan
ce
Bacterial Counts Triclosan Resistance
Field Survey • Sites sampled to date
– Non-impacted site
• Nippersink Creek, McHenry
County IL
– Heavily Impacted: Urban WWTP
• North Shore Channel
– Moderately Impacted: Suburban
WWTP
• West Branch Dupage River
• Assays completed
– Bacterial community size
– Triclosan resistance
• Assay in progress
– Microbial activity
– Respiration
– Denitrification
– Community
composition
– Triclosan
concentration
Field Survey: Next Steps
• Additional Field Sites
–Moderately Impacted: Suburban WWTP
–East Branch DuPage River
–Agricultural Sites
–Sites receiving biosolids
amendments
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