Marine ecosystems face increasing threats from chemical pollutants, particularly metals, which persist in the environment, bioaccumulate, and pose risks to marine organisms and human health. In Victoria, Australia, Burrunan dolphins (Tursiops australis), a critically endangered and endemic species, are particularly vulnerable due to their strong site fidelity to coastal regions like Port Phillip Bay and Gippsland Lakes—areas subject to significant urban, agricultural, and industrial runoff. These regions are among the most mercury (Hg)-polluted globally, with previous studies reporting elevated Hg levels in deceased dolphins. Despite this, the biological effects of metal exposure in live individuals remain poorly understood. Investigating molecular responses—especially gene expression—offers valuable insight into physiological stress and potential health impacts. This study aimed to assess metal accumulation in Burrunan dolphins and its potential effects on molecular pathways associated with detoxification, antioxidant defenses, immune function, and oxidative stress. Skin biopsies were collected from free-ranging dolphins using minimally invasive methods. Samples were preserved in RNAlater and stored at –80 °C for RNA extraction and subsequent quantitative real-time PCR (qPCR) analysis. Simultaneously, metal concentrations in the same samples were quantified using inductively coupled plasma mass spectrometry (ICP-MS). Results revealed significant associations between the expression of stress-related genes and specific metal levels, highlighting the potential use of these genes as molecular biomarkers for biomonitoring efforts. These findings emphasize the urgent need to reduce pollutant exposure in critical habitats and demonstrate the utility of molecular tools in marine conservation. By linking environmental contamination to sub-lethal molecular effects, this research supports targeted conservation strategies for Tursiops australis and provides a framework for assessing pollution impacts in marine mammals.

Spray adjuvants are products used with pesticides to improve their application or effectiveness. While exempt from federal pesticide registration by the US Environmental Protection Agency (US EPA), California law requires their registration as pesticides before sale or use. The California Department of Pesticide Regulation evaluated the suitability and acceptability of the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Mixtures Equation for predicting the acute oral toxicity of spray adjuvants as an alternative to requiring in vivo animal toxicity testing. The GHS Mixtures Equation estimates the acute oral toxicity (LD₅₀) of a mixture by combining individual LD₅₀ values of component ingredients. This study compared the results of the acute oral toxicity categories derived from the GHS Mixtures Equation with toxicity categories determined from in vivo oral studies for 51 mixtures. The results showed that the availability of LD₅₀ values for the relevant ingredients in the mixture limited the equation’s predictivity. The equation predicted the LD₅₀ values for 67% (N = 34) of the dataset. When these datasets were further compared against the US EPA toxicity classification system, the GHS Mixtures Equation results achieved 71% concordance, with toxicity either over-classified (6%) or under-classified (22%) by one category. Most discordant results were for spray adjuvant mixtures with LD₅₀ > 2000 mg/kg (limit test), particularly when 20% or more of the components had unknown LD₅₀s (N = 5). Finally, when compared against the US Occupational Safety and Health Administration’s GHS classification system, our results showed 82% concordance with 15% over-classified and 3% under-classified by one toxicity category (N = 34). Overall, the findings indicate that the GHS Mixtures Equation can reliably predict acute oral toxicity in low-toxicity mixtures, provided that all active ingredients have known LD₅₀s and that fewer than 20% of the components have unknown LD₅₀s.

The pervasive presence of pesticide residues in aquatic environments poses significant risks to both ecosystems and human health. In response to the growing need for environmentally sustainable analytical techniques, we present a novel in-syringe dispersive liquid–liquid microextraction (IS-DLLME) method utilizing a natural deep eutectic solvent (DES) for the extraction of selected pesticides from water samples.

In this study, the DES was synthesized from choline chloride and a hydrogen bond donor, offering a biodegradable and non-volatile alternative to traditional organic solvents. The microextraction was carried out inside a standard plastic syringe, eliminating the need for additional equipment and minimizing solvent consumption. Following extraction, analytes were separated and quantified using gas chromatography–mass spectrometry (GC-MS).

The method demonstrated excellent analytical performance, with extraction recoveries ranging from 86% to 97%, relative standard deviations below 4.5%, and limits of detection in the low µg/L range. The linearity was confirmed over a broad concentration range, and matrix effects were negligible. Application to environmental water samples confirmed the method's robustness and applicability to real-world analysis.

This work highlights the potential of combining IS-DLLME with DESs for rapid and sustainable monitoring of trace contaminants in water. The integration of green chemistry principles into sample preparation not only enhances analytical performance but also supports safer and more eco-conscious laboratory practices.

The progressive decline in pollinator populations, particularly bees, has become a pressing concern in recent years. One of the key contributors to this phenomenon is the infestation by parasitic mites such as Varroa destructor, which compromises bee health and compels apiculturists and agricultural producers to intensify the use of acaricides. As a result, residues of the previous mites can be transferred into the food supply via edible bee-derived products and agricultural commodities. Ensuring consumer safety in relation to said toxic products requires the implementation of stricter pesticide regulation and improved apicultural management, which in turn depends on robust analytical monitoring of hive products.

In this context, a gas chromatography mass spectrometry-based method was developed and validated for the simultaneous determination of seven commonly applied pesticides in Spain, atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate, specifically in royal jelly. The extraction procedure employed a mixture of hexane and isopropanol, followed by a clean-up using primary and secondary amine (PSA) as a sorbent. Chromatographic separation was achieved on a DB-5MS capillary column using a temperature gradient program. The method was validated according to current EU guidelines, evaluating parameters such as selectivity, detection and quantification limits, linearity, matrix effects, accuracy, and precision. Analysis of nine royal jelly samples revealed the presence of chlorfenvinphos in eight samples and α-endosulfan in one, with six samples exceeding the maximum residue limits set by European legislation.

Acknowledgements:

This research was supported by the National Plan for Scientific and Technical Research and Innovation 2013–2016, National Institute for Agricultural and Food Research and Technology-INIA–FEDER (Spain), grant number RTA2017-00004-C02-02. Ana Jano thanks the Spanish Ministry of Education and Vocational Training for her collaboration grant, and Adrián Fuente-Ballesteros to the University of Valladolid for his PhD contract.

Introduction: Copper (Cu) is a toxic and one of the most common heavy metals. Even at low concentrations, this pollutant negatively impacts the aquatic environment. This study aimed to characterize the mechanisms underlying Cu toxicity in the freshwater microalga Raphidocelis subcapitata at environmentally relevant concentrations.

Methods: Algal cultures were exposed to Cu (33 or 53 µg L⁻¹) in OECD medium up to 72 hours. Growth, photosynthetic pigments content, photosynthetic efficiency (by PAM fluorometry), intracellular ROS levels, antioxidant enzyme activity (SOD and CAT), lipid peroxidation (MDA levels), and membrane integrity were assessed over time.

Results: Both Cu concentrations significantly inhibited growth, with cell density reductions of 53% and 93%, respectively. At 33 µg L⁻¹, chlorophyll a level decreased transiently, with partial recovery by 72 hours. In contrast, exposure to 53 µg L⁻¹ led to sustained chlorophyll loss and photoinhibition, as indicated by a decrease in F_v/F_m. ROS and MDA levels increased significantly for both concentrations, with membrane damage for 53 µg L⁻¹ (5% of cells showed compromised membrane integrity). Enhanced catalase (CAT) activity indicates activation of antioxidant defences.

Conclusions: Exposure to environmentally relevant Cu concentrations caused a severe physiological disruption in R. subcapitata, as translated by the modification of different biomarkers of toxicity. This work provides insights into the algal response to Cu contamination and presents a global proposal of the toxicity pathway of Cu on R. subcapitata at environmental concentrations.

Tea stands as one of the most widely enjoyed beverages globally. Given the escalating environmental pollution with toxic substances such as pesticides, metals, and radioactivity, it is crucial to conduct monitoring studies on the presence of radionuclides in the environment, especially when the EU Council Regulation 2016/52/Euratom systematized the emergency limits on radionuclides in foods, including ²¹⁰Po, the most critical radionuclides from the ingestion pathway. This study widely presents the content of radiotoxic radionuclides, the alpha emitter ²¹⁰Po and the beta emitter ²¹⁰Pb, in 141 different tea brands collected from 18 countries worldwide. The concentrations of ²¹⁰Po of analyzed teas ranged from 1.64±0.09 Bq∙kg^-1 in green tea from Columbia (Valle de Cauca), up to 59.5±1.60 Bq∙kg^-1 also in green tea but from China (Zhejiang). Similarly to ²¹⁰Po, the highest concentration of ²¹⁰Pb activity was observed in green tea from China (Jiangxi), at 72.2±2.74 Bq∙kg^-1, while the lowest in the leaves of the rooibos plant (Aspalathus linearis) from South Africa (Cedarberg), which ranged 0.46±0.04 Bq∙kg^-1. All ²¹⁰Po and ²¹⁰Pb activity concentration values in examined teas are presented. The effective annual radiation doses and cancer risk resulting from consuming tea infusions were also calculated based on the measured activity concentrations. The research findings and outcomes highlight that consuming the investigated teas is safe from a radiological perspective.

The contamination, accumulation, spatial distribution, and potential health risks of cesium ¹³⁷Cs, polonium ²¹⁰Po, radiolead ²¹⁰Pb, and potassium ⁴⁰K in chanterelles collected across Poland were examined using validated methodology and gamma-ray and alpha-particle spectrometric measurements. Values of anthropogenic ¹³⁷Cs activity concentration in mushrooms were between 118 and 1647 Bq∙kg^-1 dry weight (dw), while for natural ⁴⁰K, they were from 1316 to 1895 Bq∙kg^-1 dw. The activity concentrations of ²¹⁰Po in chanterelles were between 2.23 and 8.57 Bq∙kg^-1 dw, and in forest topsoil, between 11.4 and 83.0 Bq∙kg^-1 dw, while corresponding values for ²¹⁰Pb were 1.50-6.14 and 7.74-46.1 Bq∙kg^-1 dw, respectively. The obtained results varied significantly: the highest values of activity concentrations were determined for ⁴⁰K, while the lowest were for ²¹⁰Po and ²¹⁰Pb. Studies have shown that lamellae mushrooms, such as chanterelles, may contain higher activity concentrations of polonium ²¹⁰Po and radiolead ²¹⁰Pb than tubular mushrooms, but a broader study is recommended. ¹³⁷Cs is an anthropogenic contaminant present in the environment as a result of global atmospheric fallout. An assessment of the values of annual radiation doses and cancer risk related to ¹³⁷Cs, ⁴⁰K, ²¹⁰Po, and ²¹⁰Pb consumed with chanterelle showed that ¹³⁷Cs and ²¹⁰Po induce a similar risk, but 2-3 orders of magnitude higher than that of ⁴⁰K and ²¹⁰Pb.

Microplastics, or plastic fragments smaller than 5 mm in diameter, are widespread water contaminants globally, raising international alarm due to their persistence, mobility, and toxicity. Microplastics enter into aquatic systems from primary (microbeads and plastic pellets) and secondary (fragmentation of larger plastics) sources through runoff, wastewater effluent, and atmospheric transport. Having invaded the environment, they are conveyed by flows and lodged in sediments or in food webs. The present review integrates recent findings concerning their mode of behavior within the environment, their biological consequences, and health implications for mankind. Microplastics are transportation media for unwanted chemicals—persistent organic pollutants and heavy metals—thatexaggerate their effects in the environment. Microplastics are widely ingested by aquatic life forms, precipitating obstructive disorder in the gut, dwarfing growth, inflammation, and trophic transfer. Human exposure is increasingly documented through seafood consumption, drinking water, and inhalation, though the long-term effects are largely uncharacterized. There is no international standardization of existing monitoring techniques, and thus comparative analysis and regulation are problematic. This review delineates the essential research requirements and calls for interdisciplinary collaboration, improved analytical tools, and policy changes. Mitigation of microplastic pollution is no longer an option—it is a requirement to safeguard ecosystem resilience and public health against a silent but increasing environmental hazard.

Organophosphates (OPs), commonly used pesticides to increase agricultural production with minimal labor efforts, unknowingly affect non-target animals. Amongst OPs, malathion (MAL) is a neuro- and immunotoxic and widely used. This investigation aimed to understand the multiple physiological responses in common carp exposed to MAL. In this study, acute toxicity test showed 6.75 mg/L as LC50 in 96 h, following which, a separate sublethal toxicity test was conducted (28 d), having environmentally realistic (2.618 μg/L) and higher concentrations (0.675 & 1.35 mg/L). In higher exposures, selected transcripts such as acetylcholinesterase (Ache) and component complement-3 (c3) levels significantly decreased. While, hematological parameters [(total erythrocytes (TEC), hemoglobin (Hb), hematocrit (Ht), Mean Corpuscular Hemoglobin (MCH), and Mean Cell Hemoglobin Concentration (MCHC)] reduced with an increase in MAL levels except for total leucocytes (TEC), which increased significantly (P<0.05). Further, histomorphology of the brain revealed disparaging alterations such as degenerated neurons, pyknosis, vacuolization, increased fibrous components, and focal necrosis with MAL exposure. Whereas gills showed hyperplasia, telangiectasia, and fused lamellae, and kidney revealed damaged glomeruli, increased lumen, and exudates. While the liver exhibited a shrunken central vein, thickened bile duct, and dilated portal vein, with exposure. Collectively, the study revealed that MAL is neuro- and immuno-toxic and damages the microarchitecture of the tissues. The non-judicious application of MAL leads to harmful effects, hence, usage should be monitored regularly.

The excessive use of plastic has raised concerns about exposure to nanoplastics (NPs) and bisphenol A (BPA) and their impacts on human health. This study aimed to assess a new method of detecting the in vitro effects of NPs and BPA that is sensitive yet simpler and faster compared to cytogenetic techniques such as cytokinesis-blocked micronucleus (CBMN) assay.

Individual and combined exposure of BPA and polystyrene nanoparticles (PS-NPs) was evaluated using the GP202 gastric cell line. The culture media, designated as 0h after exposure and 24h after the exposure, and cells 24h after exposure, were analyzed by Fourier transform infrared (FTIR) spectroscopy. CBMN did not reveal statistically significant differences in exposure to NPs and BPA compared with controls, though a trend towards an increase in MN, nucleoplasmic bridges (NBPs), and nuclear buds (NBUDs) was observed with combined exposure to PS-NPs and BPA. A univariate analysis of several FTIR spectral bands was performed after various spectral preprocessing methods, including baseline correction, baseline correction with normalization, second derivative, and second derivative with normalization. Several statistically different bands were observed, with significance at 1%, 5%, and 10%, both in multiple comparisons between exposure conditions and pairwise comparisons between exposure conditions. These changes were noted in spectra obtained from culture media and directly from cells. The FTIR spectrum represents the molecular profile of the culture media and cells, reflecting the metabolic state of the cells. Thus, FTIR spectroscopy proves to be a highly sensitive, rapid, and straightforward technique for detecting metabolic changes in gastric cell lines when exposed to NPs and BPA, even in conditions where no genotoxic changes are detected, as observed by CBMN.