Hematological and Hepatic Effects of Ulexite in Zebrafish

https://doi.org/10.1016/j.etap.2020.103496Get rights and content

Highlights

  • Low doses of UX did not cause any oxidative stress.

  • Low doses of UX promoted all enzyme activities in liver of zebrafish.

  • UX exhibits beneficial roles on blood and liver tissues of zebrafish at lower doses.

  • This applications did not alter the MN rates in zebrafish erythrocytes.

Abstract

The ulexite (UX), a borate mineral, is used as boron source and commonly used in various industrial processes. The hematological and hepatic effects of UX were investigated by exposing adult zebrafish to UX (5, 10, 20 and 40 mg/L) over 96 hours. The blood and liver tissues were taken at the end of the trial period then micronucleus (MN) rates, oxidative DNA damage (8-OHdG), apoptosis (Caspase-3), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), myeloperoxidase (MPO), paraoxonase (PON), arylesterase (AR) and lipid peroxidation (MDA) levels were determined. Genotoxic damage by UX occurred only at 40 mg/L in the blood MN assay. Oxidative stress, oxidative DNA damage and apoptosis in liver also occurred at this dose. Moreover, 5-20 mg/L doses led to decreases of DNA damage and apoptosis levels via promoting antioxidant system in liver tissues. UX exhibits beneficial roles on blood and liver tissues of zebrafish at relatively lower doses, which may be relevant to nutritional and medicinal industries.

Introduction

Boron (B) compounds, primarily boric acid and borax, are widely used in glass products, agriculture, cosmetics and pharmaceuticals (Ince et al., 2010). Previous studies indicate that boron compounds have positive effects on both human and animal health via improving the quality of life and supporting the immune system (Turkez et al., 2007; Geyıkoglu and Turkez, 2008; Ince et al., 2010; Alak et al., 2018a; 2019a; 2019b; 2019c; Basaran et al., 2019). Although borax (BX) is a common boron mineral in nature, colemanite (COL) and ulexite (UX) are also other important borate minerals (Helvacı and Alonso, 2000; Kızılca and Copur, 2015). UX, is sodium-calcium-hydroborate composition, has been localized in rare places in the earth's crust. The use of borates such as BX, COL and UX in animal and human nutrition can have key effects on various metabolic and physiological systems of the organism. However, the biochemical mechanism of boron (B) and B-containing compounds is not yet fully known. There are several assumptions about the effect of B on hormones, trans-membrane signaling, and its role in cell membrane functions and to act as a metabolic regulator in many enzymatic systems (Ince et al., 2010; Alak et al., 2018; 2019a; 2019b; 2019c, 2020).

There is little research on the oxidative and genotoxic effects of different boron compounds in experimental animal models and humans. Therefore, we have evaluated UX as a potential regulatory agent which to date has not been used in seafood. For this purpose, the study was conducted in the blood and liver tissues of the zebrafish. The Micronucleus (MN) test is the most popular cytogenetic damage index among environmental genotoxicity testing strategies and 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay were performed on blood tissue samples of the experimental fish for measuring genotoxicity potential by UX. In addition to this, malondialdehyde (MDA), 8-OHdG and apoptosis (Caspase-3) levels, protein oxidation status, activity of antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were determined in liver. In addition to these parameters, oxidative stress-related markers such as the paraoxonase enzyme (PON) include aryl esterase (ARE) and myeloperoxidase (MPO) enzyme levels were evaluated.

Section snippets

Trial plan and application of the chemical

Ulexite (UX; NaCa[B5O6(OH)6]·5H2O, Cas No. 1319-33-1, 99.8% purity) was provided from Eti Mine Works (Ankara, Turkey). The concentrations were organized according to Rowe et al. (1998). The experiment was designed in aquariums with four replications on the basis of static testing with adult zebrafish whose acclimation was completed in Atatürk University, Faculty of Fisheries, Zebrafish Research and Toxicology Trial Unit (Erzurum, Turkey). Control (0 mg/L) and UX application groups (5, 10, 20

Micronucleus test in zebrafish erythrocytes

The results of 5 mg/L, 10 mg/L, 20 mg/L, and 40 mg/L of UX applied groups as compared to the untreated control fish were presented in Fig. 1 and Fig. 2. There was no remarkable genotoxic action by the 5, 10 and 20 mg/L of UX treated groups in comparison to control fish. On the contrary the MN frequency was found to be higher only in the 40 mg/L of UX treated group.

Enzyme activities and lipid peroxidation level

Statistically significant (p < 0.05) alterations in enzyme activities and lipid peroxidation levels in liver tissues were observed

Discussion

Fish erythrocytes have been used since the 1950’s as sensitive indicators of chemical effects (Alak et al., 2019a; Katalay and Parlak, 2002). Micronucleus (MN) test is an accepted cytogenetic damage index in environmental geno-toxicity assessment. Indeed, MN detection allows the evaluation of the health of a particular species as well as water quality and the possible risks that it may have after consumption (Ozkan et al., 2011). Since MN can be applied to intact chromosomes, it can also detect

Conclusion

We determined that low doses of UX (5, 10 and 20 mg/L), did not cause oxidative stress or genotoxicity for zebrafish at the end of acute administration and that this agent showed some beneficial protective effects within this dosage range. Outside of this range, UX effects are likely to be progressively less beneficial and at 40 mg/L genotoxicity and oxidative stress becomes apparent in zebrafish.

CRediT authorship contribution statement

Gonca Alak: Conceptualization, Formal analysis, Investigation, Resources, Writing - original draft, Writing - review & editing. Fatma Betül Özgeriş: Formal analysis, Investigation. Aslı Çilingir Yeltekin: Formal analysis, Investigation. Veysel Parlak: Formal analysis, Resources. Arzu Ucar: Formal analysis, Resources. Ozge Caglar: Formal analysis, Investigation. Hasan Turkez: Conceptualization, Writing - review & editing. Muhammed Atamanalp: Conceptualization, Writing - original draft, Writing -

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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