Multielement analysis in the fish hepatic cytosol as a screening tool in the monitoring of natural waters

Tepić²,

et al. 2016

Environ Sci Pollut Res

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The study of metal bioaccumulation in the gills of European chub (Squalius cephalus) was conducted in September 2009 at the medium-sized rural river Sutla, characterized by agricultural and municipal type of water contamination. The concentration ranges were established for the first time in the soluble, metabolically available fractions of chub gills for 12 metals, which are environmentally extremely relevant and yet only seldom studied, as follows in a decreasing order: K, 225-895 mg L(-1); Na, 78-366 mg L(-1); Ca, 19-62 mg L(-1); Mg, 13-47 mg L(-1); Rb, 164-1762 μg L(-1); Sr, 24-81 μg L(-1); Ba, 13-67 μg L(-1); Mo, 1.3-16 μg L(-1); Co, 0.7-2.7 μg L(-1); Li, 0.4-2.2 μg L(-1); Cs, 0.2-1.9 μg L(-1); and V, 0.1-1.8 μg L(-1). The concentrations of Fe (1.6-6.4 mg L(-1)) and Mn (16-69 μg L(-1)) were also determined and were in agreement with previous reports. By application of general linear modelling, the influence of different abiotic (metal exposure level) and biotic parameters (fish sex, age, size and condition) on metal bioaccumulation was tested. It was established that bioaccumulation of many metals in fish depended on various physiological conditions, wherein Ba could be singled out as metal exhibiting the strongest association with one of biotic parameters, being significantly higher in smaller fish. However, it was also undoubtedly demonstrated that the concentrations of three metals can be applied as reliable indicators of metal exposure even in the conditions of low or moderate water contamination, such as observed in the Sutla River, and those were nonessential elements Li and Cs and essential element Fe. The results of our study present an important contribution to maintenance of high ecological status of European freshwaters, through enrichment of knowledge on the bioaccumulation of various metals in gills of European chub as frequently applied bioindicator species in monitoring of water pollution.

Section: Determination Of Metal Concentrations In the Soluble Gill Frmentioning

confidence: 99%

Accumulation of metals relevant for agricultural contamination in gills of European chub (Squalius cephalus)

Tepić²,

et al. 2016

Environ Sci Pollut Res

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“…Soluble cytosolic hepatic fractions, i.e. supernatants obtained after centrifugation of tissue homogenates at 50,000×g, contained cytosolic biomolecules, lysosomes and microsomes, and excluded cell membranes, nuclei, mitochondria and granules (Bonneris et al, 2005;Dragun et al, 2013a;.…”

Section: Tissue Homogenization and Isolation Of Soluble Cytosolic Tismentioning

confidence: 99%

“…During liver homogenization, an aliquot of each homogenate was set aside for subsequent digestion. Digestion procedures used in this study were modified from previously described procedures (Dragun et al, 2013a;Filipović Marijić et al, 2013). Hepatic homogenates were digested by addition of oxidation mixture (v/v 1:3), which contained concentrated HNO3 (Rotipuran  Supra 69%, Carl Roth GmbH + Co. KG, Germany) and 30% H2O2 (Suprapur  , Merck, Germany) (v/v 3:1).…”

Section: Preparation Of Hepatic Homogenates and Cytosolic Fractions Fmentioning

confidence: 99%

“…Cytosolic fractions from the first two sampling campaigns (April and September 2015) were only diluted with Milli-Q water and acidified with HNO3 (suprapur, Merck, Germany; final acid concentration in the samples 0.65%) prior to measurement. Dilution factor was 100 for Na, K, and Mg, and ten for the remaining elements (Dragun et al, 2013a). Cytosolic fractions from third and fourth sampling campaign (October 2015 and May 2016) were digested in duplicate by addition of oxidation mixture (v/v 1:1), which contained concentrated HNO3 (Rotipuran  Supra 69%, Carl Roth GmbH + Co. KG, Germany) and 30% H2O2 (Suprapur  , Merck, Germany) (v/v 3:1).…”

Section: Preparation Of Hepatic Homogenates and Cytosolic Fractions Fmentioning

confidence: 99%

“…Since the relationship between metal/metalloid concentrations and several intrinsic factors of the fish can present a confounding factor when using aquatic animals as biomonitors of metal pollution (Linde et al, 1998), we have also tested the seasonal changes of metal/metalloid concentrations in S. trutta liver, as well as their association with S. trutta sex and size. Additionally, with the aim to assess metabolically available and potentially toxic fractions of metals/metalloids in S. trutta liver, we have calculated the proportions of each metal/metalloid present in the cytosolic hepatic fractions, which contain heat-stable and heatsensitive biomolecules, lysosomes and microsomes (Bonneris et al, 2005;Dragun et al, 2013a). Finally, our overall aim was to evaluate, based on the all gathered information within this study, the current quality status of the Krka River, and the potential threat for the aquatic organisms inhabiting its water.…”

Section: Introductionmentioning

confidence: 99%

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Total and cytosolic concentrations of twenty metals/metalloids in the liver of brown trout Salmo trutta (Linnaeus, 1758) from the karstic Croatian river Krka

Marijić

Ecotoxicology and Environmental Safety

et al. 2018

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Total and cytosolic concentrations of twenty metals/metalloids in the liver of brown trout Salmo trutta (Linnaeus, 1758) were studied in the period from April 2015 to May 2016 at two sampling sites on Croatian river Krka, to establish if river water contamination with metals/metalloids downstream of Knin town has influenced metal bioaccumulation in S. trutta liver. Differences were observed between two sites, with higher concentrations of several elements (Ag, As, Ca, Co, Na, Se, Sr, V) found downstream of Knin town, whereas few others (Cd, Cs, Mo, Tl) were, unexpectedly, increased at the Krka River spring. However, total metal/metalloid concentrations in the liver of S. trutta from both sites of the Krka River were still mainly below previously reported levels for pristine freshwaters worldwide. The analysis of seasonal changes of metal/metalloid concentrations in S. trutta liver and their association with fish sex and size mostly indicated their independence of fish physiology, making them good indicators of water contamination and exposure level. Metal/metalloid concentrations in the metabolically available hepatic cytosolic fractions reported in this study are the first data of that kind for S. trutta liver, and the majority of analyzed elements were present in the cytosol in the quantity higher than 50% of their total concentrations, thus indicating their possible availability for toxic effects. However, the special attention should be directed to As, Cd, Cs, and Tl, which under the conditions of increased exposure tended to accumulate more within the cytosol. Although metal/metalloid concentrations in S. trutta liver were still rather low, monitoring of the Krka River water quality and of the health status of its biota is essential due to a trend of higher metal/metalloid bioaccumulation downstream of Knin town, especially taking into consideration the proximity of National Park Krka and the need for its conservation.

Distribution of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements among protein fractions from hepatic cytosol of European chub (Squalius cephalus L.)

Erk

et al. 2012

Environ Sci Pollut Res

Association of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements with cytosolic proteins of different molecular masses was described for the liver of European chub (Squalius cephalus) from weakly contaminated Sutla River in Croatia. The principal aim was to establish basic trace element distributions among protein fractions characteristic for the fish living in the conditions of low metal exposure in the water. The fractionation of chub hepatic cytosols was carried out by size exclusion high performance liquid chromatography (SE-HPLC; Superdex™ 200 10/300 GL column), and measurements were performed by high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Elution profiles of essential elements were mostly characterized by broad peaks covering wide range of molecular masses, as a sign of incorporation of essential elements in various proteins within hepatic cytosol. Exceptions were Cu and Fe, with elution profiles characterized by sharp, narrow peaks indicating their probable association with specific proteins, metallothionein (MT), and ferritin, respectively. The main feature of the elution profile of nonessential metal Cd was also single sharp, narrow peak, coinciding with MT elution time, and indicating almost complete Cd detoxification by MT under the conditions of weak metal exposure in the water (dissolved Cd concentration ≤0.3 μg L(-1)). Contrary, nonessential metal Pb was observed to bind to wide spectrum of proteins, mostly of medium molecular masses (30-100 kDa), after exposure to dissolved Pb concentration of ~1 μg L(-1). The obtained information within this study presents the starting point for identification and characterization of specific metal/metalloid-binding proteins in chub hepatic cytosol, which could be further used as markers of metal/metalloid exposure or effect on fish.