Microplastics (MPs) are increasingly recognized as widespread contaminants in aquatic environments, including remote freshwater systems, where their presence is linked to waste generation and transport pathways. This study examined the occurrence, spatial-temporal distribution, polymer composition, and ecological risks of MPs in surface sediments from Black Lake and Devil Lake, two high-altitude glacial lakes in Durmitor National Park, Montenegro. Sediment samples were collected across three seasons and analyzed using standardized methods, including density separation, visual identification, and polymer characterization. MPs abundance averaged 5.1 ± 1.4 items per 100 g of dry sediment in Black Lake and 3.8 ± 0.5 items per 100 g in Devil Lake. Fibers and fragments were dominant morphotypes, with particles sized 1-3 mm prevailing. Blue particles were most frequent. Five polymer types were identified, with polyethylene as the dominant polymer. Pollution load index values indicated moderate contamination, while polymer hazard and ecological risk indices suggested high to very high environmental risk. The presence of MPs in protected, high-altitude glacial lakes highlights their vulnerability to diffuse pollution sources, including tourism and long-range transport. The findings provide baseline data for alpine freshwater environments and underline the importance of integrating MPs pollution into waste management and environmental protection strategies.

Microplastic (MP) pollution in aquatic ecosystems has become a significant environmental concern worldwide. This study investigates the presence of MP in the main tributaries of the Morača river (Sitnica, Ribnica and Cijevna), the largest river flowing through the capital of Montenegro, aiming to enhance understanding of the presence, distribution, sources, and transport of MP in the Morača river basin. The present study will be covering the entire Morača river basin, yielding crucial data on MP contamination. The MP concentration in the studied rivers varied between mean values of 28.3 ± 12.2 MP/100 g dry sediment for the Sitnica, 24.9 ± 8.1 MP/100 g dry sediment for the Ribnica, and 27.3 ± 14.1 MP/100 g dry sediment for the Cijevna. The identified MPs were mainly fragments and fibers of blue, clear and red color, 0.5–1 mm in size and mainly composed of PE and PP. The results of the pollution load index indicate that the ecological status of the Morača river basin is subject to slight MP contamination, whereas the polymer hazard index results reveal a pronounced potential for adverse ecological effects. The main contribution of this study is a new insight into MP concentration in rivers and its tributaries, where the tributaries were identified as a potential important source of MP on the Morača river. This study represents a significant step towards a comprehensive understanding of the presence, distribution, sources and transport of MP pollution in the entire Morača river basin in Montenegro. The findings of this study will contribute to the growing body of knowledge about MP pollution in freshwater ecosystems, informing future research and the development of effective mitigation strategies to protect the ecological health and biodiversity of the basins.

This paper positions microbiomes as pivotal to advancing the One Health framework, emphasizing their role in linking human, animal, and environmental health. Despite widespread support, implementation faces hurdles due to insufficient metrics, standardized indicators, and limited cross-sector collaboration. An interdisciplinary team investigates how climate change, pollution, and microbial contamination influence microbiome dynamics. Microbiome indicators like stress, resilience, and stability are proposed as proxies for health to guide evidence-based interventions. Key domains for microbiome assessment include the human and animal gut, soil, groundwater, and built environments, with food safety and water quality data serving as vital connectors. The paper introduces the nanostructurome to uncover microbial communication pathways and enable precision microbiome modulation. Integration of microbiological databases, microbiome literacy, citizen science, and social science perspectives is advocated to foster coordinated action. Ultimately, the microbiome is proposed as a strategic nexus for conceptual alignment, health indicator development, and targeted interventions.

Electronic Nicotine Delivery Systems (ENDS) liquids contain compounds deemed safe for ingestion (GRAS), yet lack inhalation safety data, posing risks, especially to adolescents. This study analyzed chemical discrepancies in ENDS via gas chromatography-mass spectrometry and surveyed Slovenian secondary students on nicotine use. One-third used nicotine, two-thirds daily, mainly for stress relief. Surprisingly, 85% of physically active users also consumed nicotine. Most focused only on nicotine levels, ignoring other ingredients. Analysis revealed cytotoxic substances when inhaled and unlisted solvents, exposing a mismatch between labeled and actual content. Findings stress the need for toxicological evaluation, clearer labeling, stricter regulation, and targeted youth education.

UV filters, commonly found in personal care products, undergo chlorination in swimming pools, forming disinfection by-products (DBPs) with potential toxicological and health impacts. This study aimed to evaluate the cytotoxic, genotoxic, and biochemical effects of three UV filters - 2-hydroxy-4-methoxybenzo-phenone (benzophenone-3 or BP3), diethylamino hydroxybenzoyl hexyl benzoate (DHHB), and 4-tert-butyl-4'-methoxydibenzoylmethane (avobenzone) - and their chlorinated by-products in human lung epithelial (A549) cells and investigate their interaction with lipid membranes under visible and UV light conditions. Parent UV filters exhibited moderate cytotoxicity at higher concentrations (BP-3: 167 μM; DHHB and avobenzone: 16.7 μM), whereas their chlorinated derivatives demonstrated enhanced cytotoxic effects, particularly 3,5-dichloro BP3 (BP3-Cl: 16.7 μM; BP3-diCl: 33.3 μM) and 2-chloro avobenzone (16.67 μM). Significant increases in glutathion-S-transferase (GST) activity were observed for 3,5-dichloro BP3 at concentrations of 74.5 μM and 166.67 μM, indicating possible oxidative stress responses, but no genotoxic effects were observed suggesting other mechanism of toxicity. Membrane interaction studies showed that 3,5-dichloro BP3 caused vesicle deformation under UVA exposure (385-400 nm), highlighting the potential of chlorinated by-products to alter membrane stability. Chlorination enhances the cytotoxic potential of UV filters, likely through oxidative stress and membrane disruption. These findings emphasize the need for further research on the environmental and health implications of UV filter DBPs, particularly in contexts of prolonged UV exposure such as swimming pool environments.

Assessing the biotechnological potential of pre-peloids begins with identifying beneficial metabolites derived from microorganisms or higher plant precursors, whose presence depends on environmental conditions during peloid formation. Therefore, this study examines the impact of distinct depositional settings on lipid biomarker distribution in the organic matter (OM) of pre-peloids from freshwater continental (Vrujci, VRU), saline continental (Rusanda, RUS; Ovca, OVC), and marine (Secovlje, SEC) environments using gas chromatography-mass spectrometry (GC-MS). The OM of the VRU sample is rich in higher-plant-derived odd long-chain n-alkane members, esters of tetracosanoic and octacosanoic acids, C29 regular sterane, retene, and aromatic triterpenoids with ursane or oleanane carbon skeleton. Mixed microalgae/bacteria and higher plant biomass comprise the OM of pre-peloids deposited under saline continental environment settings. A greater participation of microalgae/ bacteria in the OM of OVC sample is inferred from the prevalence of short-chain n-C17 alkane and the highest content of fatty acid esters, while bimodal distribution of n-alkanes, with prevalence of long-chain homologs nC27-n-C33, short-chain C21 and C22 steranes, and C27 regular sterane in the OM of RUS sample, are pointing to mixed microbial/higher plant biomass. The accumulation of short-chain even n-alkane homologs, C28 ster-2-ene and C29 ster-2-ene sterenes, along with C27 hopene and diploptene, possessing (cyano)bacterial origin, dominate the OM of the SEC sample. All the above-mentioned compounds, recognized for possessing a broad spectrum of biological activities, may contribute to the final peloid product quality and applicability.

ObjectivesComposites with copper-doped mesoporous bioactive nanospheres (Cu-MBGN) were developed to prevent secondary caries by imparting antimicrobial and ion-releasing/remineralizing properties.MethodsSeven experimental composites containing 1, 5 or 10 wt% Cu-MBGN, the corresponding inert controls (silica) and bioactive controls (bioactive glass 45S5) were prepared. The temperature rise during light curing, cross-linking density by ethanol softening test, monomer elution and their potential adverse effects on the early development of zebrafish Danio rerio was investigated.ResultsMaterials combining Cu-MBGN and silica showed the highest resistance to ethanol softening, as did the bioactive controls. Cu-MBGN composites showed significant temperature rise and reached maximum temperature in the shortest time. Bisphenol A was not detected, while bis-GMA was found only in the control materials and TEGDMA in the eluates of all materials. There was no increase in zebrafish mortality and abnormality rates during exposure to the eluates of any of the materials.ConclusionsThe composite with 5 wt% Cu-MBGN combined with nanosilica fillers showed the lowest ethanol softening, indicating the polymer's highest durability and cross-linking density. Despite the TEGDMA released from all tested materials, no embryotoxic effect was observed.

In healthcare facilities, infections caused by Staphylococcus aureus (S. aureus) from textile materials are a cause for concern, and nanomaterials are one of the solutions; however, their impact on safety and biocompatibility with the human body must not be neglected. This study aimed to develop a novel multilayer coating with poly(allylamine hydrochloride) (PAH) and immobilized ZnO nanoparticles (ZnO NPs) to make efficient antibacterial and biocompatible cotton, polyester, and nylon textiles. For this purpose, the coated textiles were characterized with profilometry, contact angles, and electrokinetic analyzer measurements. The ZnO NPs on the textiles were analyzed by scanning electron microscopy and inductively coupled plasma mass spectrometry. The antibacterial tests were conducted with S. aureus and biocompatibility with immortalized human keratinocyte cells. The results demonstrated successful PAH/ZnO coating formation on the textiles, demonstrating weak hydrophobic properties. Furthermore, PAH multilayers caused complete ZnO NP immobilization on the coated textiles. All coated textiles showed strong growth inhibition (2–3-log reduction) in planktonic and adhered S. aureus cells. The bacterial viability was reduced by more than 99%. Cotton, due to its better ZnO NP adherence, demonstrated a slightly higher antibacterial performance than polyester and nylon. The coating procedure enables the binding of ZnO NPs in an amount (<30 µg cm−2) that, after complete dissolution, is significantly below the concentration causing cytotoxicity (10 µg mL−1).

Bisphenol S (BPS) is a common, persistent, and mobile chemical found in everyday products such as thermal paper. BPS can easily enter the body by migrating from the paper to the fingers, disrupting the endocrine system by mimicking the oestrogen hormone, thus negatively influencing human health. Assessing BPS levels in daily life is of great importance. This study introduces a rapid and reliable approach for detecting BPS in thermal paper and tap water by developing an electrochemical analytical method. This method allows for in-situ, real-time measurements. We present a simple, low-cost electrochemical sensor for detecting BPS using screen-printed electrodes based on carbon (SPEC) and single-wall carbon-nanotube (SPE-SWCNT) working electrodes. BPS was detected over a wide linear range from 1 to 400 μM. The detection limits were 0.73 μM and 0.87 μM for the SPE-C and SPE-SWCNT electrodes, respectively. Good repeatability was observed for both sensors when using one electrode 16 times, which demonstrates its potential for real-time environmental monitoring. Additionally, traditional chromatographic methods, high-performance liquid chromatography with a diode-array detector (HPLC-DAD), and liquid chromatography-mass spectrometry triple quadrupole (LCMS), were incorporated to enhance analytical capabilities. HPLC-DAD achieved a detection limit of 3 nM after solid-phase extraction preconcentration, while LCMS triple quadrupole demonstrated a detection limit of 10 pM without preconcentration. Electrochemical screen-printed electrodes can be employed for on-site analysis and health-risk assessments in everyday settings, such as shops. However, for detecting very low concentrations where time is not a constraint, LCMS quadrupole remains the preferred technique.

State-of-the-art technologies based on advanced oxidation processes (AOPs) are currently being developed for the removal of environmental pollutants such as pharmaceuticals from water matrices. Such studies are very important and should include the impact of inorganic and organic matrix on the degradation process of pollutants. One of the AOPs that is in focus of investigations nowadays, is ionizing radiation. Doxazosin (DOX) is a widely used pharmaceutical for the treatment of hypertension, which can be potentially harmful due to its occurrence in the aquatic environment. This study reports the radiation-induced degradation of DOX in aqueous solution. Removal of 10 mg L-1 DOX reached almost 100 % at irradiation doses of 200 Gy regardless of the dose rate used. The effect of saturated solutions with N2, N2O, air and the addition of radical scavengers such as 2PrOH and thiourea on DOX degradation was investigated. The efficiency of degradation increased in the order: thiourea < 2-PrOH < N2 < air < N2O. The effects of pH, various inorganic ions and water matrix on DOX degradation were also studied. DOX degradation was lower in underground water than in ultrapure water. Under prolonged irradiation, mineralization of about 60 % of DOX solutions was observed based on the dissolved organic carbon (DOC) evaluation. Toxicity tests with V. fischeri luminescent bacteria showed higher toxicity of samples irradiated at 500 Gy. The main degradation products were identified using LC-HRMS and degradation pathways were proposed. Overall, irradiation technology could be a promising technique for the removal of micropollutants, such as pharmaceuticals in real water matrices.

In Secovlje Salina Nature Park, the therapeutic mud matured in the natural sedimentary environmental site. This work aimed to determine the influence of the peloid maturation process on the hydrocarbon and elemental distributions, as well as changes in morphology. For this purpose, the sample before and after maturation was examined using various methods. n-Alkanes were the most abundant among saturated hydrocarbons in both immature and mature peloid samples. The results showed that the maturation mainly influenced the change in distribution and concentration (from 378 to 1958 ppm) of n-alkanes. The organic matter (OM) of the immature peloid sample was characterized by a slight prevalence of long-chain and odd carbon-numbered n-alkanes, maximizing at n-C27. However, mature peloid's OM showed a similar share of short-, mid- and long-chain n-alkanes with a slight dominance of short-chain members, maximizing at n-C16. The origin of short-chain and even carbon-numbered n-alkanes was attributed to microbial precursors (e.g., Leptolyngbyaceae). Hopanes were considerably more dominant compared to steranes in both peloids. The hopane series of immature peloid was characterized by the dominance of 22,29,30-trinor-hop-5(6)-ene (C27 hopene), as well as the presence of C30-hop-22(29)-ene (diploptene), which are widespread in cyanobacterial species. The aromatic fraction of immature peloid pointed to the predominance of polycyclic aromatic hydrocarbons (PAHs). As peloid aging progressed, the sample was richer in methyl-branched alkanes, carboxylic acids, their methyl esters, and thermodynamically more stable hopanes and steranes.

Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× g contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm–2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× g. The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.

Peptides released on frogs’ skin in a stress situation represent their only weapon against micro-organisms and predators. Every species and even population of frog possesses its own peptidome being appropriate for their habitat. Skin peptides are considered potential pharmaceuticals, while the whole peptidome may be treated as a taxonomic characteristic of each particular population. Continuing the studies on frog peptides, here we report the peptidome composition of the Central Slovenian agile frog Rana dalmatina population. The detection and top-down de novo sequencing of the corresponding peptides was conducted exclusively by tandem mass spectrometry without using any chemical derivatization procedures. Collision-induced dissociation (CID), higher energy collision-induced dissociation (HCD), electron transfer dissociation (ETD) and combined MS3 method EThcD with stepwise increase of HCD energy were used for that purpose. MS/MS revealed the whole sequence of the detected peptides including differentiation between isomeric Leu/Ile, and the sequence portion hidden in the disulfide cycle. The array of the discovered peptide families (brevinins 1 and 2, melittin-related peptides (MRPs), temporins and bradykinin-related peptides (BRPs)) is quite similar to that of R. temporaria. Since the genome of this frog remains unknown, the obtained results were compared with the recently published transcriptome of R. dalmatina.

Sample preparation is the key step in determining low concentrations of pollutants from food, biological, plant, industrial, and environmental matrices [...]

This work summarizes our research on synthesis, characterization and toxicity of selected UV-A filters and their antioxidant shield in commercial formulation - resveratrol. Benzophenone type of UV filters react under disinfection conditions with chlorine and form different mono- and dichlorinated products, while dibenzoylmethane derivatives, such as avobenzone, react with chlorine and form two main bridge chlorinated products followed by numerous chlorinated species at the advanced stages of the process. Resveratrol showed three main susceptible centers to chlorination, starting from the electrophilic addition to the double bond and continuing with the chlorination of the phenolic moieties. Several experiments conducted under different disinfection conditions (pool/sea water, addition of salts, irradiation) showed basically similar chlorination patters with some variations in terms of product formation. The results of toxicity assessment using different test organisms (Vibrio fischeri, microalgae, daphnids) have shown different sensitivity of testing organisms to the parent UV filters in comparison with chlorinated products as well as different toxicity for specific UV filter in comparison to the others. As the closing loop of all experiments in the laboratory, an up-scaling to the real human skin is presented.

Benzotriazole and its derivatives (BTAs) are commonly present in wastewater due to their extensive use in industrial processes, yet their removal is still unexplored. Here, we test the removal of these pollutants using two functionalised biochars, synthesised from wild plum (WpOH) and apricot (AsPhA) kernels. The aim of this work was to optimise the adsorption process against various BTAs (i.e., benzotriazole (BTZ), 4-hydroxy-1H-benzotriazole (OHBZ), 4-methyl-1H-benzotriazole (4 MBZ), 5-methyl-1H-benzotriazole (5 MBZ), 5-chloro-1H-benzotriazole (ClBZ), 5,6-dimethyl-1H-benzotriazole (DMBZ)), and determine the adsorption mechanisms at play, using real wastewater matrices. Batch studies showed that the optimal adsorption pH ranged between 4 and 6 for WpOH and AsPhA, respectively, and equilibrium was reached after 240 min. The kinetic models that best described the adsorption process were in the following order: Elovich model > pseudo-second order model > pseudo-first order model. The equilibrium data showed the highest correlation with the Freundlich isotherm, indicating multilayer adsorption. The maximum adsorption capacity obtained in mixtures was 379 mg/g on WpOH and 526 mg/g on AsPhA. The mechanistic work revealed that the BTAs became bound to the biochar primarily through H-bonding, n-pi and pi-pi EDA interactions. In wastewater, obtained before and after conventional treatment, the concentration of OHBZ and BTZ was reduced by >40%, while the concentration of the other compounds studied fell below the detection limit (similar to 2.0-90 ng/L). Finally, using a Vibrio fischeri assay, we showed that adsorption onto AsPhA significantly reduced the relative toxicity of both raw and treated wastewater.

The marine environment has a significant impact on life on Earth. Organisms residing in it are vital for the ecosystem but also serve as an inexhaustible source of biologically active compounds. Herein, the biodiversity of two brown seaweeds, Dictyota dichotoma and Dictyota fasciola from the Adriatic Sea, was evaluated. The aim of the study was the determination of differences in compound composition while comparing their activities, including antioxidant, antimicrobial, and enzyme inhibition, in connection to human digestion, dermatology, and neurological disorders. Chemical analysis revealed several terpenoids and steroids as dominant molecules, while fucoxanthin was the main identified pigment in both algae. D. dichotoma had higher protein, carbohydrate, and pigment content. Omega-6 and omega-3 fatty acids were identified, with the highest amount of dihomo-γ-linolenic acid and α-linolenic acid in D. dichotoma. Antimicrobial testing revealed a dose-dependent inhibitory activity of methanolic fraction against Escherichia coli and Staphylococcus aureus. Moderate antioxidant activity was observed for both algae fractions, while the dietary potential was high, especially for the D. fasciola dichloromethane fraction, with inhibition percentages of around 92% for α-amylase and 57% for pancreatic lipase at 0.25 mg/mL. These results suggest that Dictyota species might be a potent source of naturally derived agents for obesity and diabetes.

Limonene (1-methyl-4-(1-methylethenyl)-cyclohexene) is one of the most widespread monocyclic terpenes, being both a natural and industrial compound. It is widely present in the environment, including in water supplies. Therefore, it may be subjected to aqueous chlorination at water treatment stations during drinking water preparation. Besides, being a component of numerous body care and cosmetic products, it may present at high levels in swimming pool waters and could also be subjected to aqueous chlorination. Laboratory experiments with aqueous chlorination of D-limonene demonstrated the prevalence of the conjugated electrophilic addition of HOCl molecule to the double bonds of the parent molecule as the primary reaction. The reaction obeys the Markovnikov rule, as the levels of the corresponding products were higher than those of the alternative ones. Fragmentation pattern in conditions of electron ionization enabled the assigning of the structures for four primary products. The major products of the chlorination are formed by the addition of two HOCl molecules to limonene. The reactions of electrophilic addition are usually accompanied by the reactions of elimination. Thus, the loss of water molecules from the products of various generations results in the reproduction of the double bond, which immediately reacts further. Thus, a cascade of addition-elimination reactions brings the most various isomeric polychlorinated species. At a ratio of limonene/active chlorine higher than 1:10, the final products of aqueous chlorination (haloforms) start forming, while brominated haloforms represent a notable portion of these products due to the presence of bromine impurities in the used NaOCl. It is worth mentioning that the bulk products of aqueous chlorination are less toxic in the bioluminescence test on V. fischeri than the parent limonene.

Benzophenone (BP) type UV filters are common environmental contaminants that are posing a growing health concern due to their increasing presence in water. Different studies have evidenced the presence of benzophenones (BP, BP-1, BP-2, BP-3, BP-4, BP-9, HPB) in several environmental matrices, indicating that conventional technologies of water treatment are not able to remove them. It has also been reported that these compounds could be associated with endocrine-disrupting activities, genotoxicity, and reproductive toxicity. This review focuses on the degradation kinetics and mechanisms of benzophenone-type UV filters and their degradation products (DPs) under UV and solar irradiation and in UV-based advanced oxidation processes (AOPs) such as UV/H2O2, UV/persulfate, and the Fenton process. The effects of various operating parameters, such as UV irradiation including initial concentrations of H2O2, persulfate, and Fe2+, on the degradation of tested benzophenones from aqueous matrices, and conditions that allow higher degradation rates to be achieved are presented. Application of nanoparticles such as TiO2, PbO/TiO2, and Sb2O3/TiO2 for the photocatalytic degradation of benzophenone-type UV filters was included in this review.

Skin secretion of amphibians often represents the only weapon of these species against pathogens and predators. Peptides constitute the major portion of active molecules of that weapon and may be treated as potential pharmaceuticals for future generations. The first step of their efficient use involves establishing of their primary structure, i.e., sequencing. De novo sequencing by means of mass spectrometry was applied to Rana arvalis species, collected in the spring 2021 in Central Slovenia (vicinity of Ljubljana). HPLC-ESI-HRMS/MS with Orbitrap instruments was used to establish the skin peptidome of these species and compare it with the earlier identified skin peptidome of the Moscow population of Rana arvalis. Application of CID, HCD, ETD, and EThcD enabled detecting and sequencing 18 peptides; five of them were novel and may be treated as possible biomarkers of the Ljubljana population of Rana arvalis. Interestingly, representatives of two peptide families (temporins and brevinins 2) were not found in the Moscow population. MS3 modes, first of all EThcD, demonstrated their great potential in the de novo sequencing, including extraction of the sequence information from the intact peptides with disulfide cycle (rana box) in their structure and differentiation of isomeric Leu/Ile residues. Thus, all six isomeric residues were reliably distinguished in the novel melittin-related peptide AK-23-1. In addition, another post-translational modification dealing with carbonylation of the N-terminal Gly of novel temporin AVa was established using the MS3 mode. The obtained results demonstrate the efficiency of the use of MS3 tools in proteomics/peptidomics.