Acetylcholinesterase activity in muscle tissue of Norway lobster Nephrops norvegicus: Importance of body size, season, sex and naturally occurring metals.

The aim of the present study was to determine the levels of acetylcholinesterase (AChE) activity in the tail muscle tissue of wild populations of Nephrops norvegicus from the Northern Adriatic, and correlate it to body size, seasons, sex and the content of mercury, arsenic, cadmium, lead and copper. The animals of both sexes were collected in spring and autumn from two relatively distant fishing grounds. A marked variability of muscle AChE activity was found (0.49 to 11.22 nmol/min/mg prot.), displaying the opposite seasonal trend between two sampling sites. Small, but significant negative correlation has been found between AChE activity and carapace length (rs = - 0.35, p < 0.05). Data reported here provide an essential baseline for future studies of neurotoxicity in crustaceans. The study highlights the necessity for continuous monitoring of potentially toxic metals in edible marine species to avoid possible repercussions of seafood consumption on human health.

Arsenic (As), copper (Cu), zinc (Zn) and selenium (Se) in northwest Croatian seafood: A health risks assessment.

This study focuses on the health risk assessment of arsenic (As), copper (Cu), zinc (Zn), and selenium (Se) concentrations in seafood species commonly consumed in the northwestern region of Croatia. By measuring the concentrations of these elements coupled with data on seafood consumption, the health risks were evaluated using Target Hazard Quotients and Hazard Indexes. The results indicate a slightly increased health risks linked to seafood consumption for As, Cu and Zn in some of the tested seafood species. The findings of this study highlight the critical need for stronger food quality control measures, especially targeting certain types of seafood.

Assessment of health risks associated with heavy metal concentration in seafood from North-Western Croatia.

The following study aims at assessing the health risks associated with the consumption of the most commonly consumed seafood in the north-western part of Croatia due to the presence of heavy metals. Samples of seafood were collected and analysed for lead (Pb), cadmium (Cd), and mercury (Hg) content. Analyses of Cd and Pb were carried out by inductively coupled plasma mass spectrometry (ICP-MS) whereas Hg content was measured using atomic absorption spectrometry (AAS). Metal concentrations were in the following order Hg > Pb > Cd for the gilthead seabream, European hake, sardines, and tuna fish whereas in the Patagonian squid cadmium (Cd) was the heavy metal with the highest concentration, with the order of other metals being Cd > Hg > Pb. The heavy metal concentrations have been used to address the health risks using the Estimated Weekly Intake (EWI), Target Hazard Quotients (THQ), and Hazard Indices (HI). The findings revealed that the concentrations of the tested heavy metals, expressed on a per wet weight basis, did not exceed the Maximum Residue Levels (MRL) for those compounds mandated by national Croatian legislation. However, the HI for Hg was above 1, indicating a risk of adverse health effects due to the presence of this heavy metal in the consumed seafood. We conclude that the consumption of certain type of seafood such as the tuna fish should be limited when sensitive segments of the population such as children, elderly and pregnant women are concerned. Our results strongly advocate for a more stringent seafood quality control in the region.

Those That Remain: Sorption/Desorption Behaviour and Kinetics of the Neonicotinoids Still in Use.

In January 2023, the derogation loophole was closed on "emergency authorisations" for the use of three out of five neonicotinoids in all EU states. In this study, we analysed the sorption/desorption behaviour and kinetic parameters of acetamiprid and thiacloprid, the two neonicotinoids that are still approved for use, either regularly or under emergency authorisations in the EU, and widely used worldwide. Sorption and desorption curves in four soils with different organic matter content were analysed using four kinetic models, namely, Lagergren's pseudo first-order model, two-site model (TSM), Weber-Morris intraparticle diffusion model and Elovich's model. Kinetic parameters were correlated to soil physico-chemical characteristics. To determine the mutual influence of soil characteristics and sorption/desorption parameters in the analysed soils, a factor analysis based on principal component analysis (PCA) was performed. Even though the two insecticides are very similar in size and chemical structure, the results showed different sorption/desorption kinetics. The model that best fits the experimental data was TSM. Thiacloprid showed a more rapid sorption compared to acetamiprid, and, in all soils, a higher proportion sorbed at equilibrium. Intra-particle diffusion seemed to be a relevant process in acetamiprid sorption, but not for thiacloprid. Desorption results showed that acetamiprid is more easily and more thoroughly desorbed than thiacloprid, in all soils. The kinetic behaviour differences stem from variations in molecular structure, causing disparate water solubility, lipophilicity, and acid-base properties.

Tissue-specific calcium and magnesium allocation to explain differences in bulk concentration in leaves of one-year-old seedlings of two olive (Olea europaea L.) cultivars.

Olive tree (Olea europaea L.) leaves have recently been recognised as a valuable source in cosmetic and pharmaceutical industry as well as in preparation of health-supporting beverages. Little is known about the element composition of olive leaves and almost nothing about tissue-specific allocation of elements. Element composition and tissue-specific distribution were determined in leaves of two olive cultivars, Leccino and Istarska bjelica using micro-particle induced X-ray emission (micro-PIXE). In leaves of the Istarska bjelica cultivar larger bulk concentrations of potassium, sodium, molybdenum and boron, but smaller concentrations of calcium and magnesium were found than in leaves of the Leccino cultivar. Tissue-specific investigation revealed that larger concentration of calcium in epidermis and in leaf blade tissues (secondary veins, palisade and spongy mesophyll) contributed to the larger leaf bulk calcium concentration in the Leccino cultivar. For magnesium, all leaf tissues, except the bundle sheath cells and consequently the main vascular bundle, contributed to the larger bulk concentration in the Leccino cultivar. Potassium was not predominant in any of the leaf tissues examined, while sodium and molybdenum were below the limit of detection, and boron not detectable by micro-PIXE. The results indicate that sinks for calcium and magnesium are stronger in specific leaf tissues of the Leccino than of the Istarska bjelica cultivar. The new understanding of tissue-specific allocation of elements in leaves of olive will serve as a basis for detailed studies into the effects of foliar and/or soil fertilisers in olive.

Deposition of heavy metals in biological tissues of workers in metal workshops.

Welding and cutting of metals produce large amounts of particulate matter (PM), which poses a significant health risk to exposed workers. Appropriate biological markers to estimate exposure are of great interest for occupational health and safety. Here, hair and nail samples from metal workers were analyzed, which appear to be more suitable than blood or urine samples for assessing long-term exposure. Four workshops working with steel components were included in the study. The hair and nail samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) to measure the concentrations of 12 elements. At the workplaces, the concentrations of 15 elements in particulate matter were determined using X-ray fluorescence (XRF) and particle-induced X-ray emission (PIXE) techniques. The hair and nail samples of the workers contained significantly higher metal concentrations than the analytical results of a nonexposed control group. The most significant difference between the groups was found for Ti, Mn, Fe, and Co.

Biophenolic Profile Modulations in Olive Tissues as Affected by Manganese Nutrition.

Manganese (Mn) is an essential element that intervenes in several plant metabolic processes. The olive tree, and its fruits and leaves, are known as a source of nutraceuticals since they are rich in biophenols. However, there is still a serious lack of data about biophenolic distribution in olive stems and roots under Mn fertilisation. In this context, our study aimed to examine the effects of Mn fertilisation on the biophenolic profile in the leaves, stems, and roots of the 'Istarska bjelica' olive cultivar. The experiment was set up in a greenhouse, during a period of five months, as a random block design consisting of three treatments with varying Mn concentrations in full-strength Hoagland's nutrient solution (0.2 µM Mn, 12 µM Mn, and 24 µM Mn). The obtained results indicate that the amount of Mn in the examined olive plant tissues was significantly higher under 12 µM Mn and 24 µM Mn treatments compared to that of the 0.2 µM Mn treatment. While the concentration of biophenols varied in roots depending on the compound in question, a strong positive impact of the increased Mn concentration in nutrient solution (12 µM Mn and 24 µM Mn) on the concentrations of the main biophenolic compounds was observed in stems. The concentration of oleuropein in leaves almost doubled at 24 µM Mn, with the highest Mn concentration, as compared to the 0.2 µM Mn treatment. The obtained results led to the conclusion that the supply of Mn could enhance the concentration of some biologically active compounds in olives grown hydroponically, implying a critical need for further investigation of Mn fertilisation practices in the conventional olive farming system.

Characterization of airborne particulate fractions from the port city of Rijeka, Croatia.

The aim of this work was characterization of airborne particulates in the port city of Rijeka in order to evaluate impact of ship emissions on air quality. Samples of airborne particulates were collected with a ten stages cascade impactor during two campaigns: autumn and spring. A total of 16 weekly samples were analyzed on mass concentration, ions, metals and carbonaceous species (EC, OC, WSOC). Distribution of airborne fractions showed a bimodal distribution, with two maxima: one in coarse, and other in fine fraction. Source apportionment using PMF receptor model identified six sources of airborne particulates in Rijeka: crustal, biomass burning, sea salt, traffic/metal industry, combustion/SIA and HFO burning, i.e., ship emission (contribution 3%). The contribution of ship traffic to primary emission of particulate matter, using vanadium as tracer, indicated a twofold increase for PM10 and PM2.5 relative to 2012-14. An unusual desert dust event was registered in autumn campaign.

Determination of the Variability of Biophenols and Mineral Nutrients in Olive Leaves with Respect to Cultivar, Collection Period and Geographical Location for Their Targeted and Well-Timed Exploitation.

The interactive effects of cultivar, collecting period, and geographical location on the content and composition of biophenols and macro and micronutrients in olive (Olea europaea L.) leaf were investigated. Leaves of six cultivars were collected at three periods in two locations in Croatia. The leaves of Istarska bjelica cultivar had the greatest biophenol (oleuropein) potential, especially those sampled in January and in March at the location of Pag. All the cultivars yielded leaves with the highest concentration of biophenols in March, which coincided with the pruning period. Except for high oleuropein concentration in Istarska bjelica, flavonoids were found to be most useful for differentiating olive leaves according to cultivar. Verbascoside turned out to be the most potent differentiator of collecting periods, while phosphorus and zinc turned out to be most useful for differentiating locations. Despite different agroecological conditions at the two locations, cultivar exhibited a significant effect on olive leaf nutrient composition, which was certainly causally related to that of the biophenols. The results obtained showed that it is possible to plan more well-timed and efficient exploitation of biophenols from olive leaf based on the knowledge about the interactive effects of the three studied factors.

Temporal Variation of Phenolic and Mineral Composition in Olive Leaves Is Cultivar Dependent.

In order to investigate the potential of various olive cultivars and leaf sampling times for phytochemical farming practice in Croatia, phenolic and mineral composition was determined in olive leaves of four Croatian cultivars and Italian cultivar Leccino collected at three occasions, in October 2017, January 2018, and March 2018. Istarska bjelica turned out to have the largest phytochemical potential among the investigated cultivars due to steady high oleuropein concentrations found in its leaves. The concentration of main phenolic components in Istarska bjelica leaves changed only slightly during the sampling period, suggesting the possibility of its higher capability for low air temperatures stress resistance and different metabolic response compared to the other studied cultivars. Low air temperatures increased the oleuropein level and antioxidant activity in leaves of Leccino, Oblica, Levantinka, and Drobnica cultivars, which may be of crucial phytochemical farming interest. Each of the investigated olive cultivars was characterized by a specific leaf mineral nutrient composition, which could have had a specific role in their interplay with phenols.

Growth inhibition and recovery patterns of common duckweed Lemna minor L. after repeated exposure to isoproturon.

Aquatic non-targeted organisms are more likely to be exposed to herbicides in multiple pulse events then long continuous exposure. The potential of an organism to recover between exposures has an important role in the overall effects of the toxicant. Common duckweeds show high potential for recovery after a single exposure to isoproturon. To evaluate the growth patterns and recovery potential between multiple exposures, L. minor plants were exposed to isoproturon in three repetitive 7-day treatment cycles in three time-variable exposure scenarios with equivalent time-weighted average concentrations. The growth was significantly inhibited during each exposure phase with significant cumulative effects in every subsequent treatment cycle resulting in a cumulative decrease in biomass production. However, inhibitory effects were reversible upon transferring plants to a herbicide-free nutrient solution. These results indicate that L. minor plants have a high recovery potential even after multiple exposures to isoproturon. Observed cumulative decrease in biomass production, as well as the potential for fast and efficient recovery from repeated herbicide exposure, might affect the competitiveness of L. minor in surface water communities. The observations made during each exposure period, recovery patterns, and the resulting cumulative effects over time may contribute to further development, calibration and validation of mechanistic toxicokinetic/toxicodynamic models for simulating the effects of pesticides on aquatic plants populations in the laboratory and environmental conditions.

Occurence of Ochratoxin A and Biogenic Amines in Croatian Commercial Red Wines.

Food safety is one of the main concerns in the world and in wine it depends mostly on metabolites of microbial origin. The aim of this study was to investigate the occurrence of natural contaminants, ochratoxin A and biogenic amines (cadaverine, histamine, putrescine and tyramine), in Croatian commercial red wines originating from different Croatian wine-making regions. Ochratoxin A was detected in 92.8% of samples, however its concentrations in all samples were more than 10-fold lower than the limit set by the European Union (2 µg/kg), marking these wines as safe for consumption. The frequency of occurrence and measured concentrations of ochratoxin A were higher in wines produced in southern regions with highest values obtained in wines from southern Dalmatian islands. All samples were contaminated with cadaverine and putrescine, while 88.2% and 82.7% were contaminated with histamine and tyramine, respectively. Histamine concentrations ranged from below the limit of detection to 8.5 mg/L. Higher histamine concentrations were measured in wines with higher pH values which coincided with southern geographic origin. These results reinforce the need for routine detection and quantification of biogenic amines in Croatian wines to achieve better control of vinification and minimize their formation.

Croatian Wines from Native Grape Varieties Have Higher Distinct Phenolic (Nutraceutic) Profiles than Wines from Non-Native Varieties with the Same Geographic Origin.

Croatian wines made from native (Plavac mali and Teran) and non-native grape varieties (Cabernet Sauvignon and Merlot), all grown in Croatian coastal regions, were investigated. Analyses included measurements of antioxidant activities, total phenolic contents and concentrations of non-colored phenolic compounds, chosen based on their known nutraceutical properties. Plavac mali wines were distinguished by higher antioxidant activity, total phenolic content and catechin concentrations but lower flavonol concentrations. Teran wines had higher hydroxytyrosol, myricetin and resveratrol concentrations. Merlot and Cabernet Sauvignon wines had higher flavonol concentrations (except myricetin). Canonical analysis was successful in discriminating Plavac mali from Teran wines, and both varieties were separated from non-native varieties. The results indicate distinct genetic potentials of studied varieties and enable wine authentication based on the investigated bioactive compounds.

Phytotoxicity assessment of isoproturon on growth and physiology of non-targeted aquatic plant Lemna minor L. - A comparison of continuous and pulsed exposure with equivalent time-averaged concentrations.

Phenylurea herbicides are often present in the aquatic ecosystems and may be accumulated by the non-targeted organisms and impose a negative effect on the organism and the community. This study aims to investigate and compare the effects of two different isoproturon (IPU) pulse exposure scenarios on the non-targeted aquatic plant Lemna minor with effects observed in the standard test with continuous exposure. The obtained results showed that continuous IPU treatment causes significant reduction of photosynthetic pigment concentration and proteins as well as inhibition of L. minor growth. The activities of CAT, G-POX, and APX were significantly induced to diminish the accumulation of ROS under IPU treatment, but the induction of antioxidant enzymes was not sufficient to protect the plants from herbicide-induced oxidative stress. The growth of L. minor under pulse exposure to IPU recovers fast, but pulse treatment results in significant physiological changes in treated plants. The accumulation of H2O2 and lipid peroxidation products, alongside the reduced concentration of proteins and photosynthetic pigments in pulse treatment after a recovery period, indicates that IPU causes prolonged oxidative stress in L. minor plants. The recovery potential of L. minor plants after treatment with herbicides may have an important role in maintaining the population of essential primary producers in aquatic ecosystems, but IPU-induced physiological changes could potentially have a significant role in modulating the response of the plants to the next exposure event.

A baseline study of the metallothioneins content in digestive gland of the Norway lobster Nephrops norvegicus from Northern Adriatic Sea: Body size, season, gender and metal specific variability.

Metallothioneins content was investigated in digestive gland of two wild-caught Norway lobster Nephrops norvegicus populations from the Northern Adriatic Sea, in relation to body size, season and gender. Concomitant accumulation of cadmium, mercury, arsenic, lead, chromium and manganese, reactive oxygen species concentration and energy reserves in digestive gland were also assessed. While differences between genders were not recorded, metallothioneins content seasonal trends were affected by body size. Most of parameters displayed inconsistent trends across sampling sites. Significant correlation between metallothioneins content and cadmium, arsenic and mercury concentrations was recorded only for larger lobsters. A negative correlation of reactive oxygen species concentration and metallothioneins content was observed for small, but not large lobsters. Energy reserves, in particular lipids, could considerably influence biochemical and chemical parameters variations. The present results constitute the essential baseline for future studies aimed at evaluating the N. norvegicus health in relation to metal contamination of coastal sediments.

Evaluation of the Antioxidant Capacity, Antimicrobial and Antiproliferative Potential of Fir (Abies alba Mill.) Honeydew Honey Collected from Gorski kotar (Croatia).

The paper examines the antiproliferative, antimicrobial and antioxidative effects of fir (Abies alba Mill.) honeydew honey from mountain region of Croatia (Gorski kotar) as a potential replacement for standard antibiotics and chemotherapeutic agents. Cell viability, annexin V assay and flow cytometry analysis served to analyse the antiproliferative effect on, apoptosis induction in and cell death of cancer cell lines: HeLa, MCF-7, SW620, CFPAC-1, MIA PaCa-2 and normal diploid human fibroblasts (BJ). Antimicrobial activity was tested against Staphylococcus and Acinetobacter strains by agar well diffusion and microdilution assays. The DPPH˙ assay determined the radical scavenging activity, while mathematical models helped to evaluate the kinetic data of DPPH˙ inhibition. Antiproliferative effect on all tested cell lines and the prominent effect on normal diploid human fibroblasts (BJ), colorectal adenocarcinoma (SW620, metastatic) and breast epithelial adenocarcinoma (MCF-7, metastatic) was observable. The mechanisms of antiproliferative effect included accumulation of cells in the sub-G1 phase in all tested cells and induction of apoptosis in SW620 and MCF-7 cells predominantly. The antibacterial assays showed that antibiotic-resistant strains of both bacteria, including multi-resistant strain A. baumannii ATCC® BAA-1605™, were sensitive to all tested honey samples. Radical scavenging assay suggests that antioxidants present in the honey possess different radical suppressing abilities and that they react at different rates with radicals, thereby causing two steps of reaction. The results of the study indicate that Croatian fir honeydew honey has a therapeutic potential due to the strong biological activity and can serve to protect human health.

Variations of biomarkers response in mussels Mytilus galloprovincialis to low, moderate and high concentrations of organic chemicals and metals.

The changes of acetylcholinesterase activity (AChE), metallothioneins content (MTs), catalase activity (CAT) and lipid peroxidation (LPO) were assessed after 4 days exposure of mussels Mytilus galloprovincialis to a wide range of sublethal concentrations of chlorpyrifos (CHP, 0.03-100 µg/L), benzo(a)pyrene (B(a)P, 0.01-100 µg/L), cadmium (Cd, 0.2-200 µg/L) and copper (Cu, 0.2-100 µg/L). The activity of AChE in the gills decreased after exposure to CHP and Cu, whereas no change of activity was detected after exposure to B(a)P and Cd. Both induction and decrease of MTs content in digestive gland occurred after exposure to CHP and B(a)P, while a marked increase was evident at highest exposure concentrations of Cd. The content of MTs progressively decreased of MTs with increasing concentration of Cu. CAT activity and LPO in the gills did not change after exposure to any of the chemicals. The results demonstrate different response profile in relation to the type of chemical compound, and highlight the potential implications for evaluation of biological effect of contaminants in marine environment. Furthermore, the AChE activity in the gills and MTs content in the digestive gland could be modulated by CHP and Cu at environmentally relevant concentrations indicating the potential risks of short-term transient mussels exposure that may occur due to run-off from land or accidental releases.

70-kDa heat shock cognate protein expression as a putative biomarker of heavy-metal burden in Mytilus galloprovincialis.

BACKGROUND: Heavy metals naturally occur in the marine environment and ecosystems. Due to anthropogenic influence they became common waters and coastal regions pollutants in particular where their concentrations remain hazardously high. We therefore tested a protocol for combined analysis of 6 heavy metal (Pb, Cd, Cr, Zn, Fe and Hg) concentrations in mussels Mytilus galloprovincialis collected from a coastal industrial zone (shipyard locality) and mariculture facilities in combination with expression analysis of multi xenobiotic resistance related genes and stress-related gene (HSC-70). FINDINGS: In this paper we tested a protocol for heavy-metal levels assessment by use of a highly sensitive analytical method, ICP-OES, combined with expression analysis of multi xenobiotic resistance related genes, including the stress-related gene encoding 70-kDa heat shock cognate protein on mussels (Mytillus Galloprovincialis). Mussels from the shipyard locality had higher heavy metal concentrations, except Fe. Higher metal concentrations did not influence expression of multi xenobiotic resistance related genes with exception of stress-related gene (HSC-70) encoding 70-kDa heat shock cognate protein. CONCLUSIONS: Our results indicate that mussels sampled in the industrial area have increased metal concentrations in comparison with the aquaculture locality, that are accompanied by increased transcript levels of HSC-70.