Effects of copper on an omnivorous (Astyanax altiparanae) and a carnivorous fish (Hoplias malabaricus): A comparative approach
Graphical abstract
Introduction
Copper is one of the most abundant metals in nature and as it can be released naturally from its sources through geochemical processes, it is commonly found in aquatic environments (USEPA, 2007). However, anthropogenic sources such as mining, domestic sewage, and industrial sources (e.g., production of fertilizers, fungicides, electronic equipment, antifouling paints, among others) contribute significantly to this metal contaminating aquatic ecosystems (Simonato et al., 2016). Copper is essential for the life of organisms, acting in several cellular functions, being able to act as a cofactor of antioxidant enzymes, be part of the biosynthesis of neurotransmitters, and act in cellular respiration (Grosell, 2012). However, when in high concentrations in water, it can lead to adverse effects on aquatic organisms.
In freshwater fish, acute exposure to an excess of copper ions appears to target the gills, mainly causing osmoregulatory disorders and even histological damage and/or animal death (Mazon et al., 2002). Copper can lead to a decrease in Na+ uptake both by competing for input channels and by triggering an increase in the efflux of this ion in the gills and, consequently, loss of Cl− ions (Grosell, 2012). In addition to competing with the supply of Na+ in the gills, copper can cause inactivation of ATP-dependent enzymes, such as Na+/K+-ATPase, H+-ATPase, and Ca2+-ATPase (Chowdhury et al., 2016) further contributing to the ionic imbalance in freshwater fish. On the other hand, although hyponatremia is a widely reported effect in fish exposed to copper (reviewed by Grosell, 2012), hypocalcemia and the absence of ionic disturbances in relation to Na+ have also been observed in Danio rerio exposed to this metal for 96 h (Alsop and Wood, 2011).
As it has a high oxidative potential, copper can lead to the formation of reactive oxygen species (ROS) and alter cellular homeostasis (Lushchak, 2016). Consequently, after acute exposure to this metal, fish may demonstrate an increase in the production of molecules with antioxidant capacity, such as metallothioneins (Simonato et al., 2016) and glutathione (Eyckmans et al., 2011). Antioxidant enzymes have already been reported to be activated both in gills and liver, such as catalase, glutathione S-transferase (Nunes et al., 2015), and superoxide dismutase (Simonato et al., 2016) after Cu exposure. However, the reduction in the activity of these enzymes has also been observed after fish exposure to copper (Gopi et al., 2019). When antioxidant defenses are not sufficient to ensure redox balance in cells, copper can induce oxidative stress in fish, causing enzymatic inactivation, DNA damage, lipoperoxidation, and protein carbonylation (Braz-Mota et al., 2016; Simonato et al., 2016).
The response mechanisms to copper toxicity seem to be species specific in fish and are mainly associated with the ability to deal with oxidative stress, metallothionein homeostasis, and the ability to maintain ionic balance (Braz-Mota et al., 2018; Shekh et al., 2019, 2020). Freshwater fish from Neotropical regions present great diversity in ecological, morphological, and physiological attributes and their sensitivity to metals is little explored when compared to widely studied fish from temperate environments, such as Oncorhynchus mykiss and Pimephales promelas (Giacomin et al., 2018). Thus, characterizing species-specific sensitivity using biomarkers commonly used to evaluate the toxicity of metals such as copper, is extremely important for the knowledge on the toxicity of these compounds for Neotropical fauna.
Considering that fish play an important ecological role in food chains, since they transfer energy from lower basal trophic levels to higher levels, we chose to study the effects of copper on two native species, separately; one omnivorous fish, Astyanax altiparanae, and one carnivorous fish, Hoplias malabaricus. The Neotropical fish A. altiparanae (lambari) is an opportunistic species (Bennemann and Shibatta, 2002) that actively forages at different trophic levels and can thus come into contact with various contaminants. This species has been shown to be sensitive to several substances, constituting an excellent biological model for ecotoxicological studies (Vieira et al., 2014). A. altiparanae plays an essential role in the food chain of aquatic ecosystems, serving as food for carnivorous fish such as H. malabaricus, which on the other hand, is essentially a piscivorous predator with an ambush strategy (sit-wait) (Bennemann and Shibatta, 2002). H. malabaricus is mainly used in ecotoxicological studies on trophic exposure, due to its voracious behavior, high capacity to adapt to experimental conditions, and as a top chain predator (Monteiro et al., 2013).
Thus, bearing in mind the specificity of response to copper in freshwater teleosts, A. altiparanae and H. malabaricus were exposed to the same concentrations of this metal, in order to compare the response mechanisms of each species, emphasizing the effects on osmoregulation and metabolism, as well as biomarkers of oxidative stress and neurotoxicity. We hypothesized that A. altiparanae would be more sensitive to copper, since in nature, in general, mortality in the first levels of the trophic chain is higher than in top predators.
Section snippets
Acclimation
Adult females of A. altiparanae (n = 64; 11.27 ± 0.12 cm; 19.43 ± 0.61 g; mean ± SE) and juveniles of H. malabaricus (n = 64; 18.78 ± 0.64 cm; 97.80 ± 10.26 g; mean ± SE) were purchased from fish farms in the region (Paraná, Brazil). The animals were acclimated for 10 days in 500 L tanks, containing dechlorinated water, with constant aeration (> 75% saturation), 12 h light:12 h dark photoperiod, pH (~ 7.2), and controlled temperature (~ 25 °C). During the acclimation period, specimens of A.
Copper and Na+ in the water
The concentrations of total copper in the water deviated from 3.1% to 22.9% from the nominal values, with the largest variations found for the Cu 20 μg L−1 groups. Despite this, in experiments with both species, there was a concentration gradient between experimental groups (Cu 20 > Cu 10 > Cu 5 > CTR) (Table 1). The values for total copper and dissolved copper were close (Table 1), with the concentrations of dissolved copper corresponding to approximately 82% of the total copper. In addition,
Discussion
In the present study, A. altiparanae was more sensitive to copper than H. malabaricus, confirming our hypothesis. In H. malabaricus, only osmoionic disorders and increased glycemia were observed, with no increase in the concentration of copper in the tissues or oxidative stress. On the other hand, A. altiparanae seems to have lost its osmoregulatory capacity and in addition to physiological alterations, the copper concentration increased in different organs, antioxidant defenses seem not to
CRediT authorship contribution statement
Angélica Alves de Paula: Methodology, Validation, Formal analysis, Investigation, Data curtion, Writing – original draft. Wagner Ezequiel Risso: Methodology, Validation, Formal analysis. Claudia Bueno dos Reis Martinez: Methodology, Validation, Resources, Writing – review & editing, Supervision, Funding acquisition.
Declaration of Competing Interest
None.
Acknowledgements
This work is part of the PhD thesis of A. A. de Paula and was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001, in association with the Araucária Foundation of Support for Scientific and Technological Development of Paraná. Claudia B. R. Martinez is a research fellow of Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Process 307146/2019–7).
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