Environmentally relevant concentrations of tramadol and citalopram alter behaviour of an aquatic invertebrate

2. Environmentally relevant concentrations of
tramadol and citalopram alter
behaviour of an aquatic invertebrate
M. Buřič⁎, K. Grabicová, J. Kubec, A. Kouba, I. Kuklina, P. Kozák, R.
Grabic, T Randák
Present by : Sumaiah Alghamdi
Submitted to :
Dr. Proomi

3. Content
• Introduction
• Goal
• Material and methods
• Results
• Discussion
• Conclusions

4. Introduction
Pharmaceutically active compounds (PhAC) are an important
group of pollutants that represent a serious threat to aquatic
ecosystems worldwide .
• Freshwater ecosystems are exposed to mixtures of PhACs and their
residues originating from wastewater from which they are removed
only partially or not at all by sewage treatment.
• Pharmaceutically active compounds are designed to be effective at
low concentrations and their residues accumulation in the
environments can therefore affect non-target organisms .

5. Introduction…cont
• Psychoactive pharmaceuticals are designed to impact signal
pathways in the brain, so they may be expected to affect
organism behavior.
• Exposed aquatic animals often exhibit alterations that affect
predation, social interactions, reproduction, and migration.
• Assessment of the effects of PhACs requires a method simple
enough to be repeated in precisely-defined conditions as well as a
suitable model species such as crayfish.

6. Introduction…cont
• Crayfish are susceptible to behavioral changes induced by
extraneous substances in water.
• Crayfish show a complex morphology, development, and behavior,
including elaborate social interactions. They are considered
keystone species in freshwater ecosystems and are strong
ecosystem engineers. Hence, pollution impact on native
crayfish stock can result in ecosystem instability.

7. Crayfish

8. Introduction…cont
• The marbled crayfish (Procambarus virginalis ) is an ideal
model organism for many research because They are
1. Easily cultured with suitable size
2. Exhibit individuality, tolerance to handling.
3. High fertility and relatively short generation time
4. Adaptability to a wide spectrum of environmental and nutritional
conditions.
5. Marbled crayfish are genetically identical (due to reproduction by
obligatory apomictic parthenogenesis )

9. The goal of the study
• To compare the behavior of size matched, genetically
uniform, marbled crayfish exposed to environmentally
relevant concentrations of an opioid painkiller Tramadol
and the antidepressant drug Citalopram with un-exposed
controls.

10. Material and methods
• Chemicals
Tramadol hydrochloride and citalopram hydrobromide. Individual stock
solutions were prepared in ultra-pure water at a concentration of 10 mg L1
and stored at 4 °C. The exposure solutions of 1 μg L−1 were prepared by
dilution of the stock solution with aged tap water.
• Experimental animals
Marbled crayfish were cultured in the laboratory where all experimental work
was conducted. Young mature marbled crayfish specimens were randomly
selected from the culture tanks.

11. Material and methods…cont
Experimental design
Crayfish were exposed to either tramadol (7 d) or citalopram (21 d) at the
concentration of ∼1 μg L−1 of pure compound .a level considered
environmentally relevant
Experimental set-up and data acquisition
• Crayfish were placed individually in 280mm diameter plastic tanks containing
2 L aged tap water and 200 mL fine sand (< 1 mm). For each tested
compound, two groups each comprising 20 crayfish, one group without
shelter and the other with shelter Each group was further separated into 10
exposed and 10 controls..
• The crayfish were video-recorded for 4 h using a digital video to detect
patterns of crayfish movement. Distance moved (cm), activity and
velocity were evaluated. When shelters were included, the software was
programmed to detect the percentage of time spent outside the shelter.

12. Experimental set-up and data acquisition

13. Results:
With shelterwithout shelter
distance moved did not differ
significantly between exposed and
control
moved significantly shorter
distances than control animalsDistances
no significant differences in
velocity
showed significantly lower
velocity than controlsVelocity
showed no significant differences ,
both group spent less time outside
the shelter
slightly greater than controlActivity
Tramadol

14. Results…cont:
With shelterwithout shelter
the distance moved did not differ
significantly
moved significantly shorter
distances than control animalsDistances
The difference in velocity of
treated and control animals
was not significant,
showed significantly lower velocity
than controlVelocity
did not differ , Exposed and
control animals spent similar
proportion of time outside the
shelter
Activity of treated and control
animals did not differ
Activity
Citalopram

15. Discussion
• Research into the effects of PhACs on non-target aquatic organisms has
recently expanded from investigations of the most and the less commonly
occurring compounds . At least some have been shown to induce behavioral
changes in aquatic organisms at low and environmental concentrations
• Studies have confirmed that some PhACs induce behavioral alterations
(Backhaus, 2014; Fong and Ford, 2014). For example, changes in activity and
feeding rate are reported as effects of antidepressants or psychiatric drugs
(Backhaus, 2014; Brodin et al., 2014).

16. Discussion…cont
• Citalopram is described as reducing aggressive and impulsive behavior
(Armenteros and Lewis, 2002; Reist et al., 2003)
• Similar results showed that Citalpram effect the Movement of experimental
animals was significantly slower, resulting in shorter distances moved in the
absence of shelter.
• Several fish species respond to very low concentrations of Citalopram,
increased locomotor activity was observed in three spined stickleback
Gasterosteus aculeatus at 1.5 μg L−1 (Kellner et al.,2016).
• Tramadol is reported to cause decreased alertness, drowsiness, dizziness, loss
of appetite, and impaired sensitivity to environmental cues (Langley et al.,
2010).

17. Discussion…cont
• The effects of environmentally relevant concentrations of tested
compounds are clear. Concentrations of ≤ 1 μg L1 of these widely used
PhACs have shown impact on basic behavioral patterns of freshwater
biota, here represented by the crayfish.
• The tested PhACs can significantly alter behavior of large aquatic
invertebrates at dilute concentrations similar to those detected in environmental
conditions (Grabic et al., 2012; Grabicova et al., 2015;
Grabicova et al., 2017).

18. Conclusions
This research highlight the ecological importance of PhACs in aquatic
ecosystems, and call for new test protocols to examine the full environmental
impact of PhAC residues. Also presents a relatively simple method for
assessing effects on organisms of compounds commonly present in surface
waters.
Aquatic organisms living in contaminated environments are exposed
not only to single contaminants but to combinations of PhACs that could lead
to antagonistic, additive, or magnified effect. Hence, research is needed to
assess effects of particular pharmaceuticals along with studies of effects of
PhAC combinations to evaluate the real ecological impact on affected
ecosystems.