Toxicity of naproxen sodium and its mixture with tramadol hydrochloride on fish early life stages
Introduction
The occurence and fate of pharmaceuticals in water bodies have received considerable attention in recent years (Hernando et al., 2006, Verlicchi et al., 2012, Duran-Alvarez et al., 2015). These active compounds occur in surface water due to their incomplete removal during waste water treatment processes.
Naproxen and its water soluble sodium salt are members of the a-arylpropionic acids group of non-steroidal anti-inflammatory drugs (NSAIDs), known for the anti-inflammatory, analgesic and antipyretic properties (Isidori et al., 2005). NSAIDs are weak acids acting by reversible or irreversible inhibition of one or both isoforms of the cyclooxygenase enzymes, cyclooxygenase-1 (COX-1) and COX-2, involved in the synthesis of different prostaglandins from arachidonic acid (Vane and Botting, 1998). A cyclooxygenase enzyme similar to human COX-2 has been found in fish thereby making them a potential target for aquatic contamination (Zou et al., 1999). Naproxen is one of the frequently detected pharmaceuticals in water bodies. The residues of naproxen were found in Canadian sewage treatment plant (STP effluents) at levels up to 7.6 μg/L, whereas it's residues in surface waters were upto 65 ng/L (Verenitch et al., 2006). Similar levels of naproxen residues were also found by Hernando et al. (2006) in river water in Germany (70 ng/L). The levels of naproxen residues in STP influents in Sweden were upto 3.7 μg/L and in effluents upto 2.5 μg/L (Bendz et al., 2005). Considerable values of naproxen residues (up to 11 μg/L) were found by Verlicchi et al. (2012) in hospital wastewaters in Italy.
Tramadol hydrochloride and naproxen sodium are used in both human and veterinary medicine for the relief of acute and chronic pain (Vazzana et al., 2015). However, its mode of action is different from NSAIDs. Tramadol hydrochloride is a synthetic, centrally acting analgetic drug related to codeine and morphin, showing opioid and non-opioid properties (De Leo et al., 2009). It possesses a weak agonist action to mu opioid receptors and inhibits the reuptake of serotonin and norepinephrine (Raffa et al., 1992). Its occurence in water bodies has been described by various authors. Kasprzyk-Hordern et al. (2009) found average concentrations of tramadol in raw sewage in United Kingdom to be > 30 μg/L. In contrast, Wick et al. (2009) found tramadol at a concentration of 0.24 μg/L in STP influent and 0.23 μg/L in efluent in Germany.
As we reported in our previous studies (Zivna et al., 2013, Zivna et al., 2015, Stancova et al., 2014, Chromcova et al., 2015), the critical development of tissues and organs in embryos and larvae can be easily disrupted by exposure to toxic compounds. However, there is still a lack of information concerning the subchronic effects of widely used drugs on non-target aquatic vertebrates, as well as their possible mixture toxicity. The aim of this study was to investigate the effect of naproxen sodium (NS) on the early life stages of fish and to assess its mixture toxicity with tramadol hydrochloride (TH), as chemical mixtures occurring in surface bodies can be more toxic than single substances because of chemical interactions (Celander, 2011). Deleterious effects on populations are often difficult to detect in feral organisms since many of these effects tend to manifest only after longer periods of time (van der Oost et al., 2003). When the effect finally becomes clear, the destructive process may have gone beyond the point where it can be reversed. This is why, besides assessing morphometric and condition characteristics, histology, morphological anomalies and mortality, biomarkers of oxidative stress have also been assessed in this study as an early-warning signal in this study.
Section snippets
Experimental protocol
Two embryo-larval toxicity tests with common carp (C. carpio) were performed according to OECD guideline 210 (Fish, Early-life Stage Toxicity Test) (OECD, 1992). Eggs were obtained from Rybnikarstvi Pohorelice a.s. (Czech Republic), where they were produced according to standard methods of artificial reproduction (Kocour et al., 2005). The eggs were used for exposures just after fertilization and the experiment was terminated after 32 days.
In the first test, four groups were exposed to NS at
Hatching
Hatching began on day 3 post fertilization (pf) and was completed by day 6 pf in all the groups in both early-life stage tests (ELST).
In the case of the NS toxicity test the hatching was retarded on day 3 pf (p < 0.01) in all the exposed groups compared to the control. Hatching retardation was also observed on day 4 pf at concentrations of 10; 50 and 200 μg/L (p < 0.01) and at concentration of 100 μg/L of NS (p < 0.05).
No significant differences in hatching rate were observed after subchronic
Discussion
The occurence of pharmaceuticals in water bodies and their effects on aquatic vertebrates have received considerable attention in recent years (Bartoskova et al., 2013). Despite the fact that various authors have studied the effects of many NSAIDs on aquatic vertebrates (David and Pancharatna, 2009, Pamplona et al., 2011, Stepanova et al., 2013, Zivna et al., 2013, Zivna et al., 2015, Stancova et al., 2014), the effect of the frequently used analgesic NS on fish early-life stages (ELS) is not
Conclusion
The subchronic exposure to the NS and NSTH mixture had a strong effects on the ELS of common carp (C. carpio). Hatching, developmental rate, morphology, histopathology and in the case of NSTH also mortality were influenced. The bioindicators of oxidative stress were also influenced. The LOEC was determined at 10 μg/L for both NS and NSTH. Our study demonstrates that NS and NSTH exposure can adversely affect ELS of fish in terms of mortality, hatching, development, morphology and cause oxidative
Acknowledgement
This research was supported by project IGA VFU Brno 226/2015/FVHE. The authors would like to thank to Mr. Charles du Parc for English correction.
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