In this case study pharmaceuticals were analysed in the Mondego river (Portugal) and their environmental risk assessed by means of risk quotients based on an extensive retrieval of ecotoxicological data for freshwater and saltwater species. The Mondego river crosses Coimbra, the most populated city in the Portuguese Centro Region hosting a complex of regional hospitals. Environmentally relevant and prioritised pharmaceuticals were investigated in this study and their potential hazards were evaluated by conducting a separate risk assessment for the freshwater and estuary parts of the examined river section. A target analysis approach with method detection limits down to 0.01 ng L-1 was used to determine pharmaceuticals. Twenty-one prioritised target analytes out of seven therapeutical classes (antibiotics, iodinated X-ray contrast media (ICM), analgesics, lipid reducers, antiepileptics, anticonvulsants, beta-blockers) were investigated by applying ultra-high pressure liquid chromatography coupled to a triple quadrupole mass spectrometer equipped with an electrospray ionisation source. The relative pattern of pharmaceuticals along the middle to the lower Mondego showed a quite uniform picture while an approximately 40fold increase of absolute concentrations was observed downstream of the wastewater treatment plant (WWTP) discharge of Coimbra. The most frequently measured substance groups were the ICM, represented by the non-ionic ICM iopromide (βmin: 3.03 ng L-1 - βmax: 2,810 ng L-1). Environmentally more critical substances such as carbamazepine, diclofenac, and bezafibrate, with concentrations up to and 52.6 ng L-1, 59.8 ng L-1, and 10.2 ng L-1 respectively, may potentially affect aquatic wildlife. Carbamazepine revealed elevated risk quotients (RQs >1) along the middle and lower Mondego with a maximum RQ of 53 downstream of Coimbra. Especially for saltwater species, carbamazepine and clarithromycin pose high potential risks. Especially in periods of low water discharge of the Mondego river, other pharmaceuticals as diclofenac and bezafibrate may pose additional risks downstream of the WWTP.

Millions of lakes worldwide freeze, yet the fate of per- and polyfluoroalkyl substances (PFAS) in ice in freshwater systems is poorly understood. We quantified concentrations of 36 PFAS, dissolved organic carbon (DOC), and inorganic ions in ice and water in seven freshwater lakes to investigate the preferential exclusion of ions during freezing. PFAS concentrations in ice are typically lower than in the water column, demonstrating that these chemicals are excluded from ice as it freezes. However, there is preferential partitioning of both PFAS and DOC relative to cations with average sodium-normalized enrichment factors (EF) ranging from 2.74 for perfluorobutanoate (PFBA; a C4 perfluorocarboxylic acid) to 4.01 for perfluorooctanesulfonate (PFOS; a C8 perfluorosulfonic acid), with a similar EF value of 4.14 for DOC. Laboratory experiments and seasonal measurements of PFAS in the water column indicate that PFAS concentrations in ice are a function of aqueous PFAS concentrations, with lower EF values observed in waters with higher PFAS concentrations. Understanding PFAS behavior in freshwater ice is important for predicting contaminant fate during winter and spring periods, with implications for exposure to PFAS during the winter and release of PFAS when ice melts in freshwater systems.

BACKGROUND: Picocyanobacteria from the genera Prochlorococcus, Synechococcus, and Cyanobium are the most widespread photosynthetic organisms in aquatic ecosystems. However, their freshwater populations remain poorly explored, due to uneven and insufficient sampling across diverse inland waterbodies.
RESULTS: In this study, we present 170 high-quality genomes of freshwater picocyanobacteria from non-axenic cultures collected across Central Europe. In addition, we recovered 33 genomes of their potential symbiotic partners affiliated with four genera, Pseudomonas, Mesorhizobium, Acidovorax, and Hydrogenophaga. The genomic basis of symbiotic interactions involved heterotrophs benefiting from picocyanobacteria-derived nutrients while providing detoxification of ROS. The global abundance patterns of picocyanobacteria revealed ecologically significant ecotypes, associated with trophic status, temperature, and pH as key environmental factors. The adaptation of picocyanobacteria in (hyper-)eutrophic waterbodies could be attributed to their colonial lifestyles and CRISPR-Cas systems. The prevailing CRISPR-Cas subtypes in picocyanobacteria were I-G and I-E, which appear to have been acquired through horizontal gene transfer from other bacterial phyla.
CONCLUSIONS: Our findings provide novel insights into the population diversity, ecology, and evolutionary strategies of the most widespread photoautotrophs within freshwater ecosystems. Video Abstract.

The worldwide contamination of aquatic ecosystems by plastics is raising concern, including their potential impacts on the base of the food chain, which has been poorly documented. This study sought to examine, for the first time, the presence of nanoplastics (NPs) in biofilms from freshwater streams/rivers. They were collected at selected polluted sites, such as the industrial sector for plastic recycling and production, miscellaneous industries, agriculture, municipal wastewaters/effluents and road runoffs. In parallel, the functional properties of sampled biofilms were determined by proteins, lipids, esterase (lipase), viscosity and oxidative stress. The results revealed that biofilms collected at the plastic industries and road runoffs contained the highest NP levels based on size exclusion chromatography, fluorescence detection and a new nanogold sensor visualization method. Examination of the chromatographic elution profiles showed increased abundance and size of NPs in the 10-150 nm size range at the polluted sites. Biofilms from the plastic industry site had elevated levels of aldehydes (oxidative stress) and lipids compared to the other sites. Biofilms collected at the municipal sites had elevated levels of proteins and esterases/lipases, with a decrease in total lipids. Biofilms collected at agriculture sites had the lowest levels of NPs in this campaign, but more samples would be needed to confirm these trends. In conclusion, biofilms represent an important sink for plastics in freshwater environments and display signs of distress upon oxidative stress.

In a baseline study, we investigated microplastic contamination in fish from the Pinang and Kerian Rivers in Northwest Peninsular Malaysia. In recognition of the growing concern over microplastic pollution in aquatic environments, we aimed to assess the abundance and characteristics of microplastics ingested by various fish species. Fish samples were collected from local fishermen, followed by a digestion process using 10% potassium hydroxide (KOH). Microplastics were isolated and analyzed through visual examination and attenuated total reflectance Fourier transform infrared spectroscopy. The results revealed a high prevalence of microplastics, with Johnius borneensis and Oreochromis sp. exhibiting the highest abundance, averaging 48.6 and 42.8 microplastics/g, respectively. The predominant shapes were fibers (55.6%) and fragments (25.9%), with colors primarily transparent (48.19%) and black (30.12%). Our results indicate significant contamination levels in freshwater fish, emphasizing the need for further research and effective mitigation strategies. These findings provide crucial baseline data on microplastics in Malaysian freshwater ecosystems. Environ Toxicol Chem 2024;00:1-10. © 2024 SETAC.

An overview of freshwater fish variety worldwide and the variables influencing trends in variation between and within river basins are given in this review. Continental freshwater ecosystems are highly diverse and species-rich, housing nearly 18,000 species of fish (=50% of all fish species) in <0.5% of the total land area and providing a negligible (<0.01%) share of the planet\'s water supply. Large lowland tropical river basins such as the Amazon, Congo, and Mekong basins are home to the greatest freshwater fish diversity. Freshwater species of fish depth variation at the global magnitude is correlated with the total amount and variation of aquatic habitats and the environment\'s equilibrium overtime during the evolution of scales. The river continuum concept states that there is a predictable shift in fish species depth, diversity of species, and functional characteristics along gradients of environment from headwater to estuary. The ongoing trade of minerals and organic matter related to nearby floodplains is a strong factor in the number and variety of riverine fishes in most parts of the world (the flood pulse concept). Without coordinated conservation efforts, freshwater fishes will suffer significant losses in abundance and diversity due to the numerous threats they currently face worldwide. However, further development, adaptation, training, and guidance are needed. New technologies based on water conservation, suitable species, and local traditions are needed. Waste materials and local feed additives can also be used. Farmers should be provided with the necessary training and information.

The escalating production of synthetic plastics and inadequate waste management have led to pervasive microplastic (MP) contamination in aquatic ecosystems. MPs, typically defined as particles smaller than 5 mm, have become an emerging pollutant in freshwater environments. While significant concern about MPs has risen since 2014, research has predominantly concentrated on marine settings, there is an urgent need for a more in-depth critical review to systematically summarize the current global efforts, knowledge gaps, and research priorities for MP monitoring in freshwater systems. This review evaluates the current understanding of MP monitoring in freshwater environments by examining the distribution, characteristics, and sources of MPs, alongside the progression of analytical methods with quantitative evidence. Our findings suggest that MPs are widely distributed in global freshwater systems, with higher abundances found in areas with intense human economic activities, such as the United States, Europe, and China. MP abundance distributions vary across different water bodies (e.g., rivers, lakes, estuaries, and wetlands), with sampling methods and size range selections significantly influencing reported MP abundances. Despite great global efforts, there is still a lack of harmonized analyzing framework and understanding of MP pollution in specific regions and facilities. Future research should prioritize the development of standardized analysis protocols and open-source MP datasets to facilitate data comparison. Additionally, exploring the potential of state-of-the-art artificial intelligence for rapid, accurate, and large-scale modeling and characterization of MPs is crucial to inform effective strategies for managing MP pollution in freshwater ecosystems.

Properly managing aquatic organisms is crucial, including protecting endemic species and controlling invasive species. From a circular economy perspective, the sustainable use of aquatic species as a source of bioactive molecules is an area that is increasingly being explored. This includes the use of non-edible portions of seafood, which could pose considerable risks to the environment due to current methods of disposal. Therefore, it is of paramount importance to ensure that the exploitation of these resources does not result in the transfer of pollutants to the final product. This study analyzed two types of non-edible parts from the crayfish Procambarus clarkii: the abdominal portion of the exoskeleton (AbE) and the whole exoskeleton (WE), including the cephalothorax. These portions could potentially be utilized in the context of eradication activities regulated by local authorities. A screening analysis of four classes of pollutants, including pesticides, per- and polyfluoroalkyl substances (PFAS), phthalic acid esters (PAEs), and trace elements (TEs), was performed. The only analytes detected were TEs, and significant differences in the contamination profile were found between AbE and WE. Nevertheless, the levels recorded were comparable to or lower than those reported in the literature and below the maximum levels allowed in the current European legislation for food, suggesting that their potential use is legally permitted. In terms of scalability, the utilization of the entire non-edible P. clarkii portion would represent a sustainable solution for the reuse of waste products.

Improper disposal of household and industrial waste into water bodies has transformed them into de facto dumping grounds. Plastic debris, weathered on beaches degrades into micro-particles and releases chemical additives that enter the water. Microplastic contamination is documented globally in both marine and freshwater environments, posing a significant threat to aquatic ecosystems. The small size of these particles makes them susceptible to ingestion by low trophic fauna, a trend expected to escalate. Ingestion leads to adverse effects like intestinal blockages, alterations in lipid metabolism, histopathological changes in the intestine, contributing to the extinction of vulnerable species and disrupting ecosystem balance. Notably, microplastics (MPs) can act as carriers for pathogens, potentially causing impaired reproductive activity, decreased immunity, and cancer in various organisms. Studies have identified seven principal sources of MPs, including synthetic textiles (35%) and tire abrasion (28%), highlighting the significant human contribution to this pollution. This review covers various aspects of microplastic pollution, including sources, extraction methods, and its profound impact on ecosystems. Additionally, it explores preventive measures, aiming to guide researchers in selecting techniques and inspiring further investigation into the far-reaching impacts of microplastic pollution, fostering effective solutions for this environmental challenge.

Environmental DNA (eDNA) is widely used in biodiversity, conservation, and ecological studies but despite its successes, similar approaches have not yet been regularly applied to assist in wildlife crime investigations. The purpose of this paper is to review current eDNA methods and assess their potential forensic application in freshwater environments considering collection, transport and persistence, analysis, and interpretation, while identifying additional research required to present eDNA evidence in court. An extensive review of the literature suggests that commonly used collection methods can be easily adapted for forensic frameworks providing they address the appropriate investigative questions and take into consideration the uniqueness of the target species, its habitat, and the requirements of the end user. The use of eDNA methods to inform conservationists, monitor biodiversity and impacts of climate change, and detect invasive species and pathogens shows confidence within the scientific community, making the acceptance of these methods by the criminal justice system highly possible. To contextualise the potential application of eDNA on forensic investigations, two test cases are explored involving i) species detection and ii) species localisation. Recommendations for future work within the forensic eDNA discipline include development of suitable standardised collection methods, considered collection strategies, forensically validated assays and publication of procedures and empirical research studies to support implementation within the legal system.