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In the present study, essential and nonessential metal content and biomarker responses were investigated in the intestine of fish collected from the areas polluted by mining. Our objective was to determine metal and biomarker levels in tissue responsible for dietary intake, which is rarely studied in water pollution research. The study was conducted in the Bregalnica River, reference location, and in the Zletovska and Kriva Rivers (the Republic of North Macedonia), which are directly influenced by the active mines Zletovo and Toranica, respectively. Biological responses were analyzed in Vardar chub (Squalius vardarensis; Karaman, 1928), using for the first time intestinal cytosol as a potentially toxic cell fraction, since metal sensitivity is mostly associated with cytosol. Cytosolic metal levels were higher in fish under the influence of mining (Tl, Li, Cs, Mo, Sr, Cd, Rb, and Cu in the Zletovska River and Cr, Pb, and Se in the Kriva River compared to the Bregalnica River in both seasons). The same trend was evident for total proteins, biomarkers of general stress, and metallothioneins, biomarkers of metal exposure, indicating cellular disturbances in the intestine, the primary site of dietary metal uptake. The association of cytosolic Cu and Cd at all locations pointed to similar pathways and homeostasis of these metallothionein-binding metals. Comparison with other indicator tissues showed that metal concentrations were higher in the intestine of fish from mining-affected areas than in the liver and gills. In general, these results indicated the importance of dietary metal pathways, and cytosolic metal fraction in assessing pollution impacts in freshwater ecosystems. [ABSTRACT FROM AUTHOR]

The present field study, conducted during the spawning period (April/May) of European chub ( Squalius cephalus L.) from the Sava River in Croatia, indicates that seasonal changes of fish physiological state might cause variability in gastrointestinal metal (Cd, Cu, Fe, Mn and Zn), total cytosolic protein and metallothionein (MT) levels. During the period of fish spawning and increased metabolic activity, a significant relationship with chub hepatosomatic index was evident for Fe and Mn in gastrointestinal tissue ( r = 0.35 and 0.26, respectively) and in cytosolic fraction ( r = 0.32 and 0.41, respectively) and for Zn and Fe in the gut content ( r = 0.36 and 0.31, respectively). Total cytosolic protein and MT concentrations followed the same spatial distribution as Fe and Mn in all gastrointestinal fractions and as Zn in the sub-cellular fractions, with higher levels at upstream locations. Due to the role of essential metals in metabolic processes and gonad development, increased feeding and spawning activity in April/May resulted in higher gastrointestinal essential metal (Fe, Mn and Zn) and MT concentrations, which probably follow an increase in Zn concentrations, known as the primary MT inducer. Therefore, biotic factors should be considered as important confounding factors in metal exposure assessment, while their influence on gastrointestinal metal and protein levels should be interpreted depending on the season studied. [ABSTRACT FROM AUTHOR]

Abstract: The study presented here reports for the first time cytosolic metal and protein levels in the gastrointestinal tissue of field-collected European chub (Squalius cephalus), living in low metal-contaminated river section. In two sub-cellular fractions (untreated cytosol and heat-treated cytosol), isolated from the whole gastrointestinal tract of indigenous chubs collected during spawning (April/May 2006) and post-spawning period (September 2006), seasonal or gender related differences of trace metals and proteins were determined. In both fractions, cytosol and heat-treated cytosol, metal levels decrease as follows: Zn>Fe>Cu>Mn>Cd and are significantly higher during the spawning period. Besides that, the level of heat-sensitive proteins (metalloenzymes) in cytosolic fractions is also significantly higher during the spawning period in both male and female specimens. Higher condition indices of chubs in the spawning period imply higher feeding activity, energy reserves and recent growth of indigenous chubs. Metallothionein levels, determined in the heat-treated cytosol, are comparable in gastrointestinal tract of feral chubs collected in both seasons and confine the background metallothionein levels, on average 3mgg− 1 wet tissue. Chub spawning has been identified as a confounding factor, having an impact on cytosolic metal and protein levels, presumably due to enhanced food supply (higher condition indices) and fish spawning (higher gonadosomatic indices in some male specimens). [Copyright &y& Elsevier]