1. • Forty-eight percent of people reported taking at least one
prescription drug in 2007-2008 (Zhong et al. 2013).
• Pharmaceuticals could affect non-target organisms after
evidence of similar drug target composition to humans
(Fick et al. 2010).
• Pharmaceuticals enter the environment from human
excrement or from disposal of unused/expired drugs in the
toilet (Corcoran et al. 2010) (figure 1).
• Wastewater treatment facilities have limited ability to
remove pharmaceuticals from sewage (Lajeunesse et al.
2011).
• Citalopram is the most efficiently removed of selective
serotonin reuptake Inhibitors (SSRIs) at 98% (Styrishave et
al. 2011).
• SSRIs are a major class of pharmaceutical detected in
wastewater effluent. Serotonin reuptake transporter
(SERT) is the main target receptor for this drug class
(Kreke and Dietrich, 2008).
• SSRIs work by blocking SERT, thereby increasing the
amount of serotonin in the synapse (figure 2). Serotonin
plays many physiological purposes including controlling
appetite and modulating motor output (Brodin et al. 2014)..
• Citalopram exists as a racemic mixture with the S
enantiomer responsible for the antidepressant effects
(figure 3). A racemic mixture is used in this study.
• Citalopram is highly selective towards SERT while showing
the lowest acute toxicity of SSRIs. Citalopram is one of the
least studied of the SSRI antidepressants (Silva et al. 2015).
• Citalopram exposed three-spine stickleback showed a decrease
in feeding behavior by 38% (Kellner at al. 2014).
The purpose of our research:
• To determine if citalopram causes an effect on hybrid
striped bass ability to catch prey
We hypothesize that:
• Citalopram will cause a decrease in feeding behavior
of hybrid striped bass
Effects of citalopram on hybrid striped bass predatory behavior
Lauren Stoczynski1, Stephen Klaine1, Joseph Bisesi2
Institute of Environmental Toxicology, Clemson University [1]
Center for Environmental and Human Toxicology, University of Florida [2]
Methods
Discussion/Future WorkIntroduction
References
Results
Discussion
• Time to consume prey data only includes bass which ate prey
during the exposure period.
• Preliminary data suggests six days of exposure to citalopram
causes an increase in time to consume minnows between high,
medium, and low treatments on day 6, for the first prey.
• At a high enough concentration citalopram could bind to enough
SERT causing the sublethal effect of decreased feeding.
• The bass chosen for the exposure all consumed at least three
minnows within the first 10 minutes of the 25 minute feeding period.
• During the exposure bass in the all treatment groups showed a
significant decrease in number of minnows consumed compared to
the control (figure 6).
• The decrease or elimination of hunger could suggest that at a high
enough concentration citalopram causes hunger suppression in
bass.
• From an ecological standpoint, reduction or elimination of hunger
could cause exposed bass to be less ecologically fit for competition
compared to unexposed bass.
• Rainbow trout and guppy fish both showed a decrease in
aggressive behavior and change in sexual behavior, respectively
(Holmberg et al. 2011).
• Food consumption for trout decreased compared to controls when
exposed to citalopram. The only trout that showed a decrease in
feeding however were those that experienced dominate behavior
vs. subordinate (Lepage et al. 2005).
Future work
• Full Exposures ending on the 6th, and 3rd day still need to be
completed
• Exposures ending on the 9th day need to be completed
• HPLC work determining SSRI water concentrations
• Sample preparation and LC-MS/MS of plasma and brain samples
Brodin, T., et al., Ecological effects of pharmaceuticals in aquatic systems-impacts through
behavioral alterations. Philosophical Transactions of the Royal Society, 2014. 369.
Corcoran, J., M. Winter, and C. Tyler, Pharmaceuticals in the aquatic environment: a critical
review of the evidence for health effects in fish. Critical Reviews in Toxicology, 2010.
40(4): p. 287-304.
Kreke, N. and D.R. Dietrich, Physiological endpoints for potential SSRI interactions in fish.
Critical Reviews in Toxicology, 2008. 37: p. 215-247.
Fick, J., et al., Therapeutic levels of levonorgestrel detected in blood plasma of fish: results from
screening rainbow trout exposed to treated sewage effluents. Environmental Science
and Technology, 2010. 44: p. 2661-2666.
Holmberg, A., et al., Does waterborne citalopram affect the aggressive and sexual behavior of
rainbow trough and guppy? Journal of Hazardous Materials, 2011. 187: p. 596-599.
Kellner, M., et al., Environmentally relevant concentrations of citalopram partially inhibit feeding
in the three-spine stickleback (Gasterosteus aculeatus). Aquatic Toxicology, 2015. 158:
p. 165-170
Lajeunesse, A., et al., Distribution of antidepressants and their metabolites in brook trout
exposed to municipal wastewaters before and after ozone treatment- evidence of
biological effects. Chemosphere, 2011. 83: p. 564-571.
Lepage, O., et al., Serotonin, but not melatonin, plays a role in shaping dominant-subordinate
relationships and aggression in rainbow trout. Hormones and Behavior, 2005. 48: p.
233-242
Silva, L., et al., Reviewing the serotonin reuptake inhibitors (SSRIs) footprint in the aquatic biota:
uptake, bioaccumulation and ecotoxicology. Environmental Pollution, 2015. 197: p.
127-143.
Styrishave, B., B. Halling-Sorensen, and F. Ingerslev, Environmental risk assessment of three
selective serotonin reuptake inhibitors in the aquatic environment: a case study
including a cocktail scenario. Environmental Toxicology and Chemistry, 2011. 30(1): p.
254-261.
Zhong, W., et al., Age and Sex Patterns of Drug Prescribing in a Defined American Population.
Mayo Clinic Proceedings, 2013. 88(7): p. 697-707.
Acknowledgements
I would like to thank my mentor Steve Klaine for all he has done for me
during my two years at Clemson. I would like to thank my committee
members Joe Bisesi and Tom Schwedler for their support and ability to
answer my questions. Additionally I would like to thank all of the Klaine
lab.
Figure 2: mode of action for SSRI
class pharmaceuticals
Figure 3: Average time taken for bass to eat the first prey. *
represents the high concentration being statistically difference
from the medium and low treatment groups. Statistics were
done using JMP ANOVA and t test for comparing all mean
combinations.
Figure 4: Average time taken for bass to eat the second prey.
Statistics were done using JMP t test for comparing all mean
combinations.
Figure 5: Average time taken for bass to eat the third prey.
Statistics were done using JMP t test for comparing all mean
combinations.
http://www.the-scientist.com/?articles.view/articleNo/4
3615/title/Drugging-the-Environment/
Figure 3: enantiomers of
citalopram
http://www.buyzembrin.biz/chemistry/
2.
Bass are group trained (1.). Enough prey
are added to ensure that all bass have had
a chance to eat. Training completed three
times over three days
Bass are transferred to individual 80L tanks
with three sides of aquarium covered (2.).
Allowed to acclimate for three days before
being fed four minnows on day 0, 3, and 6.
Time to eat each prey was recorded.
On third individual training day bass who
adequately eat minnows are exposed to low,
medium, high or control treatment groups
(50, 100, and 150 µg/L). Water turned off.
Bass are fed on day 0, 3, and 6 then water
is turned back on and bass are fed on days
9 and 12 for a recovery period
Analytical techniques:
• Solid phase extraction for concentrating
SSRI in tank
• HPLC for determining SSRI
concentrations
• LC-MS/MS for determining serotonin and
SSRI concentrations in plasma and brain
samples
1.
0
50
100
150
200
250
300
0 3 6 9 12
averagetimetoeatminnow
(seconds)
Day
Control
low
medium
high
0
50
100
150
200
250
300
350
400
450
0 3 6 9 12
Averagetimetoeatminnow(seconds)
Day
control
low
medium
high
0
200
400
600
800
1000
1200
1400
1600
0 3 6 9 12
Averagetimetoeatminnow(seconds)
Day
control
low
medium
high
Figure 6: average minnow consumption per day by bass exposed to citalopram. * represent
low, medium, and high treatment group days which are significantly different from the control
associated with that treatment. ** represents a p value of 0.0503, which is just over the
significance threshold but could still suggest a decrease in feeding. Statistics were done
using student t test.
Figure 1: graphic for pharmaceutical
entrance into the environment
0
0.5
1
1.5
2
2.5
3
3.5
4
0 3 6 9 12
average#minnowsconsumed
Day
control
low
medium
high
* *
*
*
*
*
*
*
*
*
**
*