Marija Sedak

Lead and cadmium in red deer and wild boar from different hunting grounds in Croatia.

The concentration and relations of Cd and Pb as environmental risk factors were studied by atomic absorption spectrophotometry in the liver, kidney and muscle of free ranging wild boar (n=94) and red deer (n=45) from hunting grounds in four counties of north-east Croatia. In all four counties, the levels of Cd found in the kidney of red deer ranged from 2.28 to 5.91 mg/kg, and in wild boar from 3.47 to 21.10 mg/kg. The mean renal concentration of Cd was significantly higher in wild boar than in red deer from all four study areas. The mean hepatic (0.11 to 0.49 mg/kg, respectively) and muscle (0.01 to 0.04 mg/kg, respectively) Cd concentrations were similar in both species. The mean renal Cd concentration in wild boar and red deer exceeded 1 mg/kg in all four counties, ranging from 67.0% to 91.4% and from 45.5% to 69.2%, respectively. Also, the hepatic Cd/renal Cd ratio was lower than 1 in all animals. In all four counties, renal Pb concentration ranged from 0.058 to 3.77 mg/kg in red deer and from 0.056 to 11.60 mg/kg in wild boar. Hepatic Pb concentration was similar in both species (0.061 to 0.202 mg/kg in wild boar and 0.077 to 0.108 mg/kg in red deer). Because of the high Cd level in the organs of wild boar and red deer, further research is needed to identify the source of contamination in order to preserve the health of animals and humans.

As, Cd, Hg and Pb in four edible shellfish species from breeding and harvesting areas along the eastern Adriatic Coast, Croatia.

Four species of shellfish (Mytilus galloprovincialis, Ostrea edulis, Chlamis varia and Venus verrucosa) were collected during the autumn 2011 and spring 2012 along the eastern Adriatic coast from six shellfish harvesting areas (all species) and 13 breeding sampling areas (mussels) to assess As, Cd, Hg and Pb levels and the human risks of shellfish consumption. The mean metal concentrations (wet weight) in the examined species ranged from 1.420 to 9.575mg/kg for As, 0.034 to 1.270 for Cd, 0.005 to 0.680 for Hg and 0.140 to 2.072 for Pb. Examination of the spatial distribution of As, Cd, Hg and Pb revealed statistically significant differences among the studied areas. Since the concentrations were below the maximum prescribed by the laws of the EU and Croatia (the concentrations slightly exceeded the upper limits for three samples; Pb, Cd and Hg) and the hazard index, (HI) for Cd, Hg and Pb were below 1 and the target cancer risk (TR) for As was lower than 1×10(-6), there is no human health risk of consumption of shellfish from Croatian waters.

Concentrations of Trace Elements in Tissues of Red Fox (Vulpes vulpes) and Stone Marten (Martes foina) from Suburban and Rural Areas in Croatia

Trace elements concentrations (As, Cd, Cu, Pb and Hg) were determined in the liver, kidney and muscle of 28 red fox (Vulpes vulpes) and 16 stone marten (Martes foina) from suburban and rural habitats from Croatia. Rural and suburban habitats affected Cd and Hg levels in the muscle, liver and kidney of red fox. Significant differences in metal concentrations in the muscle, liver and kidney were detected among species. Suburban stone marten accumulated the highest levels of trace elements (mg/kg w.w.): in muscle 0.019 for Hg; in liver 0.161 for Cd, 36.1 for Cu and 0.349 for Pb; in kidney 1.34 for Cd and 0.318 for Pb. Values observed were higher than those found in suburban red fox and therefore, may represent an important bioindicator for the accumulation of toxic metals in urbanized habitats.

Balance of glucosinolates content under Cd stress in two Brassica species

We designed a pot experiment to study the changes of growth parameters, accumulation of Cd in cabbage and kale, during a prolonged exposure period with two harvests (after 45 and 75 days of plant growth). Simultaneously, we monitored changes of total S and GSL contents in different plant organs. Both Brassica species showed tolerance to Cd, since no obvious symptoms of metal toxicity were displayed and no significantly reduced plant biomass were recorded. Moreover, a trend of root biomass enhancement was noticed. In general, the Cd concentration in roots was higher than in other organs. Data indicate that the transport system from roots to shoots was already saturated after the first growing period, while root continued to accumulate Cd during the second growing period. Regardless of differences in the GSL content between cabbage and kale, both Brassica species had a similar response to Cd exposure, indicating that the Brassicacea plant family could have unique features for the regulation of GSL content in order to balance the roles of GSLs as defence compounds and as S reservoirs. During the first growing period, GSLs in leaves and root could be more involved in ameliorating S deficiency rather than plant defence. After long-term Cd exposure, we found differences between GSL patterns in leaves and root. Furthermore, we found increased total GSL levels in the stem during whole experiment which indicates that stem could serve as a GSLs storage organ under Cd stress. This paper provides new insight into the possible roles of GSL in Cd stress.

Survey of arsenic, cadmium, copper, mercury and lead in kidney of cattle, horse, sheep and pigs from rural areas in Croatia

Trace element (As, Cd, Cu, Hg, Pb) concentrations were determined in the kidney of cattle, sheep, horses and pigs from rural areas of Croatia. Arsenic concentrations in kidney tissues ranged from 0.013 to 0.5 mg kg−1. No significant differences in As kidney levels were observed among species. The highest levels of Cd and Hg were found in horses and ranged 0.029–47.4 and 0.009–0.13 mg kg−1, respectively. The European Union maximum levels for Cd in kidney were exceeded by 92.3% of horses, 14% of cattle and 16% of sheep. The highest mean Cu levels were found in sheep and horse (8.53 and 8.45 mg kg−1). Mean Hg concentrations in kidney of cattle, sheep, horses and pigs were 0.051, 0.011, 0.034 and 0.094 mg kg−1, respectively, and the highest levels of Pb were found in cattle (1.71 mg kg−1). Significant differences in Cd, Pb and Hg concentrations between animal species were observed.

Determination of Macro- and Microelements in Cow, Goat, and Human Milk Using Inductively Coupled Plasma Optical Emission Spectrometry

The contents of macro- and microelements in cow, goat, and human milk from Croatia were measured and compared for the first time. The highest concentrations of element were measured in milk cow: calcium 1.4 g/kg, sodium 0.6 g/kg, zinc 4.0 mg/kg, strontium 0.4 mg/kg; goat: magnesium 0.2 g/kg, potassium 2.2 g/kg, iron 0.8 mg/kg, manganese 71 µg/kg, selenium 35 µg/kg, molybdenum 20 µg/kg, chromium 72 µg/kg, lithium 14 µg/kg; and human: copper 193 µg/kg. Equal content of molybdenum (20 µg/kg) was determined in cow and goat milk. Significant differences in element levels between the three species were determined. Concentrations of magnesium, manganese, selenium, chromium, and lithium in goat milk were significantly higher than in cow milk. Significantly lower levels of calcium, potassium, sodium, magnesium, zinc, selenium, and strontium were determined in human milk than in cow and goat milk. Chromium content was significantly higher in goat than in human milk. Copper content in human milk was significantly higher than in cow milk. Correlations, mostly moderate and positive, were found between elements in cow and goat milk.

Trace and macro elements in the femoral bone as indicators of long-term environmental exposure to toxic metals in European brown bear (Ursus arctos) from Croatia

We explored the long-term accumulation of aluminium, strontium, cadmium and lead in the compact and trabecular bone of the femoral epiphysis, metaphysis and diaphysis in 41 brown bears (Ursus arctos) from Croatia. Also, we assessed their influence on macro and trace elements (sodium, magnesium, phosphorus, potassium, calcium, manganese, iron, cobalt, copper, zinc and barium) in bears’ bone. There were no sex differences in element levels in general, while age was associated with bone length and levels of all elements, except for cadmium. Elements had different levels depending on the part of the bone sampled. More pronounced differences were observed between the compact and trabecular regions, with higher levels of majority of elements found in compact bone. Moderate to high associations (Spearman coefficient, rS = 0.59–0.97) were confirmed between calcium and potassium, magnesium, phosphorus, manganese, cobalt, zinc, strontium and lead. Lead levels in the bone were below those known to cause adverse health effects, but in 4 of 41 animals they exceeded baseline levels for domestic animals. The femoral bone of the brown bear reflected the accumulative nature of lead and strontium well, as it did the impairment of bone-forming essential element levels associated with these two elements. However, the distribution pattern of elements along the bone was not uniform, so additional care should be taken when choosing on the part of the bone sampled.

Essential and toxic element concentrations in monofloral honeys from southern Croatia

The concentrations of 24 elements in seven honey types (multifloral, heather, common heather, bearberry, sage, mandarin orange-blossom and honeydew) collected in southern Mediterranean regions of Croatia were determined using ICP-MS. Significant differences were found in the concentrations of Ag, As, Ba, Cu, Co, Fe, K, Mg, Mn, Mo, Na, Ni, Se, Sb, U and Th (p<0.05, all) among honeys. The highest element concentrations were determined in honeydew honeys, with the exception of multifloral (Ca, Cr, Mo, Se), common heather (Mg, Na), bearberry (Ba, Fe, Pb) and sage (Ag) honeys. Among the floral honeys, the highest concentrations were found in multifloral honey (Al, As, Be, Ca, Cr, Mn, Mo, Ni, Se, Th and U), common heather (Co, K, Mg, Na, V), sage (Ag, Cd, Cu), and bearberry (Ba, Fe, Pb, Sb, Zn). The results contribute to the evidence supporting the role of botanical origin on the elemental composition of honey.

Validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sulfonamides, trimethoprim and dapsone in muscle, egg, milk and honey

ABSTRACT A quantitative multi-residue method that includes 13 sulfonamides, trimethoprim and dapsone was developed and validated according to Commission Decision 2002/657/EC for muscle, milk egg and honey samples. For all matrices, the same extraction procedure was used. Samples were extracted with an acetone/dichloromethane mixture and cleaned up on aromatic sulfonic acid (SO3H) SPE cartridges. After elution and concentration steps, analytes were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Data were acquired according to the multiple reaction-monitoring approach (MRM) and analytes were quantified both by the isotope dilution and the matrix-matched approaches calculating the response factors for the scanned product ions. The developed method shows good linearity, specificity, precision (repeatability and within-laboratory reproducibility), and trueness. Estimated CCβ for sulfonamides ranged between 5.6 and 8.2 µg kg−1 for eggs, between 11.1 and 69.9 µg kg−1 for milk, between 64.7 and 87.9 µg kg−1 for muscle, and between 2.7 and 5.3 µg kg−1 for honey. CCβ values for dapsone were 3.1, 0.6, 0.7 and 1.5 µg kg−1 and for trimethoprim were 3.1, 6.7, 81.7 and 3.0 µg kg−1 calculated for eggs, milk, muscle and honey, respectively. Recovery for all matrices was in the range from 89.1% and 109.7%. In matrix effect testing, no significant deviations were found between different samples of muscle and milk; however, a matrix effect was observed when testing different types of honey. The validation results demonstrate that the method is suitable for routine veterinary drug analysis and confirmation of suspect samples.

Concentration of mercury and selenium concentrations in the tissues of five cetacean species from Croatian coastal waters

Mercury (Hg) and selenium (Se) concentrations were measured in muscle, liver, kidney, spleen and lung tissues of five cetacean species, three dolphin (Stenella coeruleoalba, Tursiops truncatus and Grampus griseus) and two whale species (Balaenoptera physalus and Ziphius cavirostris), stranded along the Croatian coast during the period 1999-2002. Statistically significant differences in Hg concentrations in muscle, spleen and lung, and Se in liver and lung of the different dolphin species were observed. Mercury levels in liver and spleen and Se levels in liver differed between young and adult T. truncatus species. A significant positive correlation between different tissue types for Hg and Se concentrations was observed. In all tissues tested, the lowest Hg and Se concentrations were found in B. physalus. Mercury concentrations were positively correlated with Se in all tissues. The results present one of few studies related to lung and spleen tissues in these mammals, particularly in the Adriatic Sea. Since very little data are available, this research provides new data on concentrations of Hg and Se in five cetacean species from the Adriatic Sea basin.