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The natural gas industry bears a certain contamination risk to human and biota due to, among others, mercury, arsenic, and naturally occurring radioactive material content in gas. We tracked multiple stabile metal(loid)s and radionuclides within the natural gas treatment plant Molve, Croatia, ecosystem during the last decade through a comprehensive monitoring of soil, earthworms, moss, livestock (blood, milk, hair, urine, and feces from cows), and wildlife animals (brain, muscle, liver, and kidney of European hare and pheasant). The level of mercury and other stable metal(loid)s has shown temporal variation, but without an obvious trend. The found spatial differences in soil and earthworms were based on the differing soil characteristics of the sampled locations and exceeded the maximal allowable concentration of arsenic and zinc for agricultural soil. The status of essential copper, selenium, and zinc in cows, hares, and pheasants inclined towards deficiency. The measured stable metal(loid) levels in soil and animal samples were generally in the same range of values reported in earlier decades from the same area or non-polluted areas across Europe. The consumption of local cow and game products (e.g., milk, meat) can be considered safe for human health, although game offal is advised to be avoided as a food item due to the low risk of lead and cadmium's adverse effects. Although the activity concentrations of some radionuclides in moss were higher than reported for pristine areas, transfer from soil to moss was assessed as average (except for lead-210). Radiological risk to human and biota around the gas treatment plant Molve was estimated as negligible., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

In the last two decades (1990-2012), as part of a mercury monitoring programme, earthworms and soils have been collected from four locations in the vicinity of a natural gas production and treatment plant near the village of Molve, Croatia. The aim of this study was to determine the concentration of mercury in the collected samples, monitor its changes over a longer period of time and determine the bioaccumulation of total mercury in earthworms (Eisenia fetida) from the soil. Total mercury concentrations in earthworms from the surroundings of four boreholes (Molve 9-12) ranged within 0.195-1.050, 0.129-1.0, 0.229-1.236 and 0.223-0.799 μg g -1 dry weight, while total mercury concentrations in different soil types at the same locations within 0.055-0.350, 0.035-0.250, 0.031-0.240 and 0.071-0.475 μg Hg g -1 of soil. The calculated mercury bioaccumulation factor ranged between 0.9 and 17.5. Mercury levels in soil and earthworms, as a tool for soil pollution assessment, suggested low mercury exposure and risks for human health in the monitored area.

Environmental impact of natural gas facility near Molve (Podravina, Croatia) was assessed using hares (Lepus europaeus Pallas) as biomonitors. Elevated levels of heavy metals in the environment lead to their accumulation in different tissues of hares. The authors have tested accumulation and distribution of several metals in hare liver, kidney, and muscle tissues. The accumulation of copper in hare liver and kidneys with concomitant decrease of zinc was observed in animals from Podravina region as opposed to the control group of animals (Island Krk, Croatia). Secondly, the expression of metallothioneins was assessed because of their crucial role in metal homeostasis. Observed elevation of metallothionein expression in tested organs emphasizes the possible prolonged negative effects of heavy metals in the surroundings as well as a state of oxidative stress in animals. Further monitoring of the area is necessary for better control of hydrocarbon processing to diminish the possible negative environmental effects.

Cadmium occurs naturally in the environment and as an anthropogenic pollutant. Exposure to low concentrations of cadmium is inevitable and may produce toxic effects. Another important aspect of cadmium toxicity is its interaction, often antagonistic, with essential elements such as selenium. The aim of this study was to highlight the risks of long-term exposure to low cadmium concentrations, using a scientific and chemical approach and hares (Lepus europaeus Pallas) as model organisms in a field study. Two study areas were monitored. Levels of cadmium and selenium were quantified in the organs of hares, the expression of metallothioneins I + II and the products of lipid peroxidation were determined. The median cadmium concentrations (wet weight) in the muscle, liver, kidney and brain of hares from an exposed group ranged from 0.033 to 0.037, 0.763 to 1.054, 3.090 to 16.594 and 0.016 to 0.087 µg g(-1), respectively; whereas, the median selenium concentrations (wet weight) ranged from 0.100 to 0.108, 0.153 to 0.332, 0.677 to 0.701 and 0.078 to 0.116 µg g(-1), respectively. Expression of the metallothioneins I + II proteins was observed in tissues. Lipid peroxidation (LPO) levels, measured as malondialdehyde (MDA) equivalents, increased with the cadmium concentration. Further research on long-term exposure to low concentrations of cadmium in the environment is needed., (Copyright © 2013 John Wiley & Sons, Ltd.)

The aim of this research was to assess the degree of heavy metal (cadmium, lead and mercury) contamination of wild boars from central Croatia and thereby conduct further tests to connect the contamination to oxidative stress occurrence. The highest cadmium concentration was noticed in the kidneys of older boars, the lead concentration was approximately the same in the liver and kidneys, but it was much higher than the concentration in the muscles, while the highest mercury concentration was measured in the kidneys of all wild boars. The correlation between the malondialdehyde (MDA) and the heavy metal concentrations was higher in tissue samples taken from younger animals (1 to 3 years of age) than in the tissue samples taken from older animals (5 to 6 years of age). In the kidneys of all wild boars there was a statistically significant connection between the MDA and cadmium concentration; in the kidneys of younger wild boars there was a statistically significant connection between the MDA and lead concentration. Further research on this topic should focus on MDA potential to find its use, not only as a contamination biomarker in the area of ecotoxicology, but also in the evaluation of the hygienic acceptability of animal products.

The aim of this study was to analyze and evaluate risks of long-term exposure to mercury in hares (Lepus europaeus Pallas), with a chemical-analytical approach evaluating median mass fraction of toxic mercury in the hares organs (liver, kidney, muscle and brain). To obtain better insight into possible effects of mercury, the study included screening of the oxidative status after long term exposure to low concentrations of mercury. Hares organs were analyzed for total mercury concentration by AAS. Glutathione and glutathione-dependent enzymes status was also investigated. The median mercury concentrations (wet weight) in the liver, kidney, muscle and brain of the hares ranged from 0.058-0.189, 0.138-0.406, 0.013-0.046 and 0.022-0.102 μg/g respectively. Concentration of the glutathione (GSH), glutathione-peroxidase (GPx), and glutathione-reductase (GR) activity increased with the mercury concentration. However, glutathione S-transferase (GST) and superoxide-dismutase (SOD) activity decreased with the mercury concentration. The results of this study show the impact of environmentally absorbed mercury on the antioxidant status of the examined hares. Further research on long-term exposure to low concentrations of mercury is needed.

Since 1995 the concentrations of total mercury were analyzed in the pheasant organs (liver, kidney, brain and muscle) that live freely in an area that was exposed to elevated levels of mercury as a result of industrial activity (mercury contaminated natural gas production and processing which began in 1980). The range of the median mercury concentration values (wet weight) in pheasant organs in Podravina, Croatia were 0.0004-0.024, 0.0001-0.034, 0.0002-0.016 and 0.0001-0.025 μg/g for liver, kidney, brain, and muscle, respectively. Results of these studies confirms that in the study area in the observed period there was no significant mercury contamination which can be linked to anthropogenic activities (hydrocarbon production)‥ On the contrary, systematic measurements demonstrate a small but constant (barely noticeable) decline in mercury concentrations in pheasant's organs. Primarily, this is the result of the significant improvement in the technological process which involves the removal of mercury from the natural gas that was designed and implemented in 1993. However, due to the need for reliable control of mercury contaminated natural gas production and the importance for better understanding of long-term exposure to low levels of mercury, and assessment of their possible harmful effects on the environment, human and animal health, research will be continued.

Polychlorinated biphenyl (PCB) and organochlorine pesticide (OCP) levels in fat tissue of red and fallow deer (Cervus elaphus L. and Dama dama L.) from two inland and an Adriatic area were established. Of 17 analysed PCBs, PCB-28, PCB-138, PCB-153, PCB-180 and PCB-118 were found in all samples, whilst PCB-101 and PCB-170 were found in more than 50% of samples. They ranged between 0.03 and 5.98 ng g(-1) fat weight. Of seven analysed OCPs, HCB, γ-HCH and 1,1-dichloro-2,2-di(4-chlorophenyl)ethylene (DDE) were found in all samples, whilst β-HCH was found in more than 50% of samples. They ranged between 0.17 and 22.14 ng g(-1) fat weight. The dominating compounds were DDE, PCB-138, PCB153, PCB-118 and PCB-180. According to the Duncan multistage test, the levels of PCB-138, PCB-153, PCB 170 and PCB-180 were significantly higher in perirenal fat samples of specimens taken from the Adriatic area. DDE was significantly higher in the inland deer samples. Some species differences were determined and were mostly related to higher PCB and β-HCH levels in fallow deer samples and higher DDE levels in red deer samples. No sex difference was established. As for age, significantly higher levels of PCB-118 and PCB-52 were found in fawns.

Concentrations of cadmium, lead and mercury were determined in muscle, liver and kidney tissue from three different age groups of red deer (young animals aged 6 or 7 months, middle-aged animals aged 3 to 5 years, old animals aged over 8 years) from the Baranja region of Croatia. Median cadmium concentrations were low in the muscles of all investigated age groups (0.0002; 0.0009 and 0.0020 μg/g), higher in the liver (0.0279; 0.0656 and 0.1463 μg/g) and highest in the kidneys (0.4792; 2.8531 and 6.1657 μg/g). A positive correlation was established between cadmium concentration and age. In all analyzed tissues the median lead concentration was higher in young (muscle 0.0024; liver 0.0364 and kidney 0.0618 μg/g), compared to middle-aged animals (muscle 0.0001; liver 0.0184 and kidney 0.0160 μg/g). In contrast to cadmium, mercury had a negative correlation to age but median concentrations were very low (in muscle: 0.0011; 0.0001 and 0.0006 μg/g; in liver: 0.0051; 0.0037 and 0.0022 μg/g and in kidney: 0.0145; 0.0183 and 0.0106 μg/g) especially compared to cadmium. From the hygienic point of view all the examined tissues are edible since concentrations of the analyzed metals do not exceed values proposed by the official regulations, with the exception of a few kidney samples, which contained cadmium above the recommended value. Furthermore, mercury concentrations are no longer (since 2008) a matter of legislation.

In the last two decades (1990-2008), as part of a comprehensive monitoring of the eco-system, hares were collected in the vicinity of the mercury contaminated natural gas production and treatment plant Molve, Croatia. Their organs (muscle, liver, kidney and brain) were analyzed for total mercury concentration by cold vapor AAS. The range of the median mercury concentration values (wet weight) in hares organs were 0.001-0.005, 0.007-0.045, 0.022-0.126 and 0.0006-0.015 μg/g for muscle, liver, kidney and brain, respectively. The results of mercury measurements in hares organs during the period of last twenty years demonstrate a small but constant decline in concentration values. Comparing the results obtained in this study with results published in available data and literature on mercury concentration in hare's tissue it can be concluded that area investigated in this research belongs to low mercury contaminated region. Nevertheless, further eco-monitoring and mercury measurements in various hares organs are valuable and necessary and will be continued.