Malondialdehyde concentrations in the intestine and gills of Vardar chub (Squalius vardarensis Karaman) as indicator of lipid peroxidation

Environmental Science and Pollution Research - A lipid peroxidation product, malondialdehyde (MDA), was studied in Vardar chub (Squalius vardarensis Karaman) as an indicator of oxidative stress,...     

The influence of the season and the biotic factors on the cytosolic metal concentrations in the gills of the European chub (Leuciscus cephalus L.)

The influence of the season and the biotic factors (age and gill mass) on metal and protein levels in the gill cytosol of the young chubs (2 and 3 years old) was studied in Sava River in autumn 2005 and spring 2006. The obtained results are the first reported cytosolic concentrations of five metals and proteins for the chub gills. The average levels in autumn and spring, respectively, for total cytosolic proteins were 11.2 and 19.9 mg ml(-1), for Zn 6.3 and 10.3 microg ml(-1), for Fe 3.9 and 9.6 microg ml(-1), for Cu 68.4 and 79.0 ng ml(-1), for Mn 55.0 and 63.5 ng ml(-1), and for Cd 2.9 and 3.6 ng ml(-1). The influence of the gill mass on both the protein and the metal levels was observed, but it was seasonally dependent. In autumn, positive correlations were obtained between the gill mass and four parameters (total proteins, Mn, Zn and Fe), and negative with Cu. Contrary, in spring, even negative correlations of total proteins and some metals with the gill mass were observed. The proposed explanation for the different dependence of metal levels on the gill mass in autumn and spring was the seasonal difference in feeding intensity and metabolic rate, with presumably faster metabolism and water filtration through gills in spring. This hypothesis was further supported by the statistically significantly higher concentrations of the total proteins, Zn and Fe, as well as the Fulton condition indices in the spring period.     

Assessment of low-level metal contamination using the Mediterranean mussel gills as the indicator tissue

Purpose  

The aim of this study was to compare the level of metal contamination in two bays in the middle part of the Eastern Adriatic coastal zone in Croatia using the gills of mussels Mytilus galloprovincialis as indicator tissue. Despite the existing sources of contamination, previous studies with caged mussels only indicated moderate metal contamination of the Kaštela Bay, contrary to the Trogir Bay in which marina and shipyard present a probable source of Cu- and Zn-contamination.     

Risk factors in urban road traffic accidents

INTRODUCTION: The urban road traffic accident (RTA) risks for the city of Zagreb, Croatia, from 1999 through 2000 were analyzed with the aim of reducing the increasing injury incidence. METHOD: Simple and bivariate analysis using chi(2), odds ratio, and confidence interval of 95% was used to determine risks in three outcome groups: killed, severely, and mildly injured. RESULTS: There were 528 RTA victims consisting of 260 severely, 213 mildly injured, and 55 killed at the scene of an accident and during transportation. More fatal accidents occurred during night hours (OR=3.78; 95% CI, 2.08-6.85), on urban road links (OR=2.33; 95% CI, 1.30-4.19), and at exceeding speed limit (OR=2.56; 95% CI, 1.43-4.61). More people were injured than killed on urban junctions (OR=5.27; 95% CI, 2.21-12.57). The highest combined risk of dying or being severely injured was found in males, driving at excessive speed, on urban links, and during bad visibility (OR=16.15; 95% CI, 3.901-66.881). CONCLUSION: These results will influence the urban traffic police enforcement measures, which will change inappropriate behavior of drivers and protect the least experienced road users.     

Metal and metallothionein level in the heat-treated cytosol of gills of transplanted mussels Mytilus galloprovincialis Lmk

A stock of mussels Mytilus galloprovincialis was transplanted over 1 year to four sites in a semi-enclosed bay in Croatia which is under the influence of various sources of pollution. The positive correlation of metal (Cd, Zn, Cu, Mn, Fe) and metallothionein (MT) tissue contents based on the analysis of heat-treated cytosol of gills with shell mass, as an indicator of mussel age, indicated to accumulation of metals, as well as the increase of MTs with mussel age. The principal component analysis (PCA) revealed that 74% of total variance of obtained results could be explained through two principal components. The first principal component was highly correlated with MT, Cd and Zn indices (metal or MT content/shell mass), and the second one with Cu, Mn, and Fe indices, as well as the gill index. High correlation of MTs with Zn and Cd is consistent with their affinity for binding to MTs.     

Assessment of general condition of fish inhabiting a moderately contaminated aquatic environment

The assessment of general condition of fish in the moderately contaminated aquatic environment was performed on the European chub (Squalius cephalus) caught in September 2009 in the Sutla River in Croatia. Although increases of the contaminants in this river (trace and macro elements, bacteria), as well as physico-chemical changes (decreased oxygen saturation, increased conductivity), were still within the environmentally acceptable limits, their concurrent presence in the river water possibly could have induced stress in aquatic organisms. Several biometric parameters, metallothionein (MT), and total cytosolic protein concentrations in chub liver and gills were determined as indicators of chub condition. Microbiological and parasitological analyses were performed with the aim to evaluate chub predisposition for bacterial bioconcentration and parasitic infections. At upstream river sections with decreased oxygen saturation (～50 %), decreased Fulton condition indices were observed (FCI: 0.94 g?cm^?3), whereas gonadosomatic (GSI: 2.4 %), hepatosomatic (HSI: 1.31 %), and gill indices (1.3 %) were increased compared to oxygen rich downstream river sections (dissolved oxygen ～90 %; FCI: 1.02 g?cm^?3; GSI: 0.6 %; HIS: ～1.08 %; gill index: 1.0 %). Slight increase of MT concentrations in both organs at upstream (gills: 1.67 mg?g^?1; liver: 1.63 mg?g^?1) compared to downstream sites (gills: 1.56 mg?g^?1; liver: 1.23 mg?g^?1), could not be explained by induction caused by increased metal levels in the river water, but presumably by physiological changes caused by general stress due to low oxygen saturation. In addition, at the sampling site characterized by inorganic and fecal contamination, increased incidence of bacterial bioconcentration in internal organs (liver, spleen, kidney) was observed, as well as decrease of intestinal parasitic infections, which is a common finding for metal-contaminated waters. Based on our results, it could be concluded that even moderate contamination of river water by multiple contaminants could result in unfavourable living conditions and cause detectable stress for aquatic organisms.     

The Influence of the Biometric Parameters on Metallothionein and Metal Level in the Heat-Treated Cytosol of the Whole Soft Tissue of Transplanted Mussels

The influence of the biometric parameters (shell mass, whole soft tissue mass, condition index) on MT and metal levels in the heat-treated cytosol of the whole soft tissue of transplanted mussels was studied over the period of one year. The positive correlation of three metals (Cd, Fe, Zn) with the shell mass indicated to time-dependent increase of their contents. Strong correlation of Mn content with the whole soft tissue mass (r = 0.74, p < 0.0001), and almost identical changes of these two parameters over the year make Mn a good indicator of mussel's condition. As opposed to the other metals, Cu content does not exhibit connection with biometry. On the other hand, MTs are highly influenced by biometry. As much as 65% of their variability could be explained by the changes of the shell mass and the whole soft tissue mass. Consequently, it is difficult to distinguish if the obtained positive correlation between Cd and MTs (r = 0.48, p < 0.05) reflects MT induction by Cd, or Cd accumulation as a result of age-dependent increase of MTs. Due to the strong influence of the biometry on MT level, the whole soft tissue is not considered as the best choice for measuring MTs as a biomarker. Better option would be to isolate a specific tissue that shows indisputable connection between MT induction and metal accumulation.     

Distribution of Co,Cu, Fe,Mn, Se,Zn, and Cd among cytosolic proteins of different molecular masses in gills of European chub (Squalius cephalus L.)

The distribution of essential elements Co, Cu, Fe, Mn, Se, and Zn, and nonessential element Cd among cytosolic proteins of different molecular masses in the gills of European chub (Squalius cephalus) sampled in the moderately contaminated Sutla River in September of 2009, was studied after the protein separation by size exclusion high-performance liquid chromatography (SEC-HPLC), and the metal determination in the obtained fractions by high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS). The aims of the study were to characterize the distribution profiles of metals within different protein categories in gills in the conditions of low metal exposure in the river water, and to compare them with the previously published hepatic profiles. The distribution profiles of analyzed metals were mainly characterized with several peaks. However, some observations could be emphasized: both Cu and Cd were eluted near metallothionein elution time; elution time of one of Co peaks could be associated with Co-containing compound cobalamin; increasing cytosolic Fe concentrations resulted in possible Fe binding to storage protein ferritin; both Mn and Zn had poorly resolved peaks covering wide ranges of molecular masses and indicating their binding to various proteins; both Zn and Se increased in protein fractions of molecular masses <5 kDa following their concentration increase in the gill cytosol; expected clear metallothionein peak was not observed for Zn. Comparison of gill profiles with previously published hepatic profiles revealed similar and in case of some elements (e.g., Co, Fe, Mn, and Se) almost identical distributions in both organs regarding elution times. On the contrary, heights of obtained peaks were different, indicating possible metal binding to the same proteins in the gills and liver, but in different proportions. The results obtained in this study can be used as a basis for comparison in monitoring studies, for identification of changes that would occur after exposure of chub to increased metal concentrations.     

Preliminary assessment of total dissolved trace metal concentrations in Sava River water

This study provides the preliminary data set for total dissolved trace metal concentrations in the surface water of the Sava River in Croatia and the assessment of Sava River water quality status. The highest levels of total dissolved metals were observed for Fe, Mn, and Zn (12.6 +/- 7.8 mirog L(-1), 3.44 +/- 3.95 mirog L(-1), and 2.27 +/- 1.53 mirog L(-1), respectively), the intermediate concentrations for Ni, Cu, and Cr (0.59 +/- 0.14 mirog L(-1), 0.54 +/- 0.14 mirog L(-1), and 0.32 +/- 0.06 mirog L(-1), respectively), and the lowest levels for Co, Pb, and Cd (0.064 +/- 0.022 mirog L(-1), 0.055 +/- 0.051 mirog L(-1), and 0.011 +/- 0.004 mirog L(-1), respectively). The results refer to the grab water samples taken at five sites in the period from March to June, 2006. For four trace metals (Mn, Pb, Zn, and Fe), the high temporal variability within one season was observed. It can present a problem for reliable evaluation of total dissolved concentrations of these metals in the river water, if the assessment is based on the occasional grab water sampling. The comparison of results obtained in this study with previously reported data for several unpolluted rivers indicated that Sava River water reflects a certain anthropogenic impact. However, according to the levels proposed by European regulations, it still can be classified as water containing total dissolved trace metals in concentrations not significantly above the natural level.     

Radiocaesium activity concentrations in potatoes in Croatia after the Chernobyl accident and dose assessment

Systematic investigations of (137)Cs and (134)Cs activity concentrations in potatoes (Solanum tuberosum) for the post-Chernobyl period (1986-2005) in the Republic of Croatia are summarized. The correlation between (137)Cs activity concentrations in fallout and potatoes, has been found to be very good, the correlation coefficient being r = 0.88 with P(t) < 0.001 for 18 degrees of freedom. As the radiocaesium levels in potatoes decreased exponentially, the mean residence time of (137)Cs in potatoes was estimated by fitting the measured activity concentrations to the exponential curve. The mean residence time was found to be 6.8 +/- 1.1 years, the standard deviation being estimated by the Monte Carlo simulations. The initial observed (134)Cs:(137)Cs activity ratio in potatoes has been found to be quite variable, but slightly lesser than the theoretically predicted value of 0.5, calculated by applying the known inventory of these radionuclides in the Chernobyl reactor to the equation for the differential radioactive decay. This can be explained by presence of the pre-Chernobyl (137)Cs in soil that originated from nuclear fallout. The annual effective doses received by (134)Cs and (137)Cs intake due to consumption of potatoes estimated for an adult member of the Croatian population were found to be very small, as the per caput Dose for the entire 1986-2005 period was calculated to be about 2.9 microSv, (134)Cs accounting approximately for 1/3 of the entire dose. Therefore, after the Chernobyl accident consumption of potatoes was not the critical pathway for human intake of radiocaesium from the environment in Croatia.