Assessment of parent and alkyl -PAHs in surface sediments of Iranian mangroves on the northern coast of the Persian Gulf: Spatial accumulation distribution, influence factors, and ecotoxicological risks.

Spatial patterns, potential origins, and ecotoxicological risk of alkylated (APAH) -and parent -(PPAH) polycyclic aromatic hydrocarbons (PAHs) were studied in mangrove surface sediments along the northern coasts of the Persian Gulf, Iran. The mean total concentrations (ngg-1dw) ∑32PAH, ∑PPAHs and ∑APAHs in sediments were 3482 (1689-61228), 2642 (1109-4849), and 840 (478-1273), respectively. The spatial variability was similar among these PAH groups, with the highest levels occurring in Nayband National Marine Park (NNMP). Physicochemical environmental factors, such as sediment grain size, and total organic carbon (TOC) contents, are significant factors of PAH distribution. These findings suggest that PAH pollution level is moderate-to-high, supporting the current view that mangrove ecosystems are under intensive anthropogenic impacts, such as petrochemical, oil and gas loads, port activities, and urbanization. Non-parametric multidimensional scaling (NPMDS) ordination demonstrated that NNMP mangrove is the critical site exhibiting high loading of PAH pollutants. Here, for the first time in this region, Soil quality guidelines (SQGs), Toxic equivalency quotient (TEQ), Mutagenic equivalency quotient (MEQ), and composition indices comprising Mean maximum permissible concentration quotient (m-MPC-Q), and Mean effect range median quotient (m-ERM-Q) methods were used to have a comprehensive risk assessment for PAH compounds and confirmed medium-to-high ecological risks of PAHs in the study area, particularly in the western part of the Gulf, highlighting the industrial impacts on the environment.

Chemical Activity-Based Loading of Artificial Sediments with Organic Pollutants for Bioassays: A Proof of Concept.

Persistent organic pollutants (POPs) pose a risk in aquatic environments. In sediment, this risk is frequently evaluated using total or organic carbon-normalized concentrations. However, complex physicochemical sediment characteristics affect POP bioavailability in sediment, making its prediction a challenging task. This task can be addressed using chemical activity, which describes a compound's environmentally effective concentration and can generally be approximated by the degree of saturation for each POP in its matrix. We present a proof of concept to load artificial sediments with POPs to reach a target chemical activity. This approach is envisioned to make laboratory ecotoxicological bioassays more reproducible and reduce the impact of sediment characteristics on the risk assessment. The approach uses a constantly replenished, saturated, aqueous POP solution to equilibrate the organic carbon fraction (e.g., peat) of an artificial sediment, which can be further adjusted to target chemical activities by mixing with clean peat. We demonstrate the applicability of this approach using four polycyclic aromatic hydrocarbons (acenaphthene, fluorene, phenanthrene, and fluoranthene). Within 5 to 17 weeks, the peat slurry reached a chemical equilibrium with the saturated loading solution. We used two different peat batches (subsamples from the same source) to evaluate the approach. Variations in loading kinetics and eventual equilibrium concentrations were evident between the batches, which highlights the impact of even minor disparities in organic carbon properties within two samples of peat originating from the same source. This finding underlines the importance of moving away from sediment risk assessments based on total concentrations. The value of the chemical activity-based loading approach lies in its ability to anticipate similar environmental impacts, even with varying contaminant concentrations. Environ Toxicol Chem 2024;43:279-287. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Does pre-exposure to polluted sediment affect sub-cellular to population-level responses to contaminant exposure in a sentinel species?

Understanding how key-species respond to anthropogenic stress such as chemical pollution is critical for predicting ecosystem changes. Little is however known about the intra-specific variability in the physiological and biochemical traits involved in contaminant exposure responses. Here, we explored this idea by exposing the Baltic amphipod Monoporeia affinis from two sites, one moderately polluted and one more pristine, to a sediment spiked with PAHs and PCBs. We evaluated the amphipods responses related to feeding, growth, a stress biomarker (acetylcholinesterase [AChE] inhibition) and stable isotope (δ13C and δ15N) composition including isotope niche analyses. More adverse responses were expected in animals from the low-pollution site than those from the high-pollution site due to tolerance development in the latter. Amphipods from both populations showed a ∼30% AChE inhibition when exposed to the contaminant spiked sediment. However, both controls and exposed amphipods from the high-pollution site had higher survival, nutrient uptake and condition status than the amphipods from the low-pollution site, which did not feed on the added diatoms as indicated by their isotope values. We found no signs of population-specific responses in physiological adjustments to contaminants with regard to classic ecotoxicological biomarkers such as AChE inhibition and growth status. Instead, isotope niche analyses proved useful in assessing contaminant stress responses at the population level.

Cardiotoxic and neurobehavioral effects of sucralose and acesulfame in Daphnia: Toward understanding ecological impacts of artificial sweeteners.

Artificial sweeteners are widely used in food and pharmaceuticals, but their stability and persistence raise concerns about their impact on aquatic life. Although standard toxicity tests do not reveal lethal effects, recent studies suggest a potential neurotoxic mode of action. Using environmentally relevant concentrations, we assessed the effects of sucralose and acesulfame, common sugar substitutes, on Daphnia magna focusing on biochemical (acetylcholinesterase activity; AChE), physiological (heart rate), and behavioural (swimming) endpoints. We found dose-dependent increases in AChE and inhibitory effects on heart rate and behaviour for both substances. Moreover, acesulfame induced a biphasic response in AChE activity, inhibiting it at lower concentrations and stimulating at higher ones. For all endpoints, the EC50 values were lower for acesulfame than for sucralose. Additionally, the relationship between acetylcholinesterase and heart rate differed depending on the substance, suggesting possible differences in the mode of action between sucralose and acesulfame. All observed EC50 values were at µg/l levels, i.e., within the levels reported for wastewater, with adverse effects observed at as low as 0.1 µg acesulfame /l. Our findings emphasise the need to re-evaluate risk assessment thresholds for artificial sweeteners and provide evidence for the neurotoxic effects of artificial sweeteners in the environment, informing international regulatory standards.

Aggregation in experimental studies with microparticles: Experimental settings change particle size distribution during exposure.

The ubiquitous occurrence of microplastics is raising broad concerns and motivating effect studies. In these studies, however, particle behaviour in the water and aggregation are rarely considered leading to contradictory results reported by different studies. Using an environmentally relevant experimental setup with Daphnia magna as a test organism, we investigated how experimental conditions affect particle aggregation and the aggregate heterogeneity in terms of the particle size distribution. The experimental factors considered were (1) exposure duration (48 h vs 120 h), (2) the total mass of suspended solids (0-10 mg/l) composed of natural mineral particles (kaolin) and microplastics, (3) the proportion of the microplastics in the particle suspension (0-10% by mass), (4) dissolved organic matter (DOM; 0 vs 20 mg agarose/l), and (5) presence of the test organism (0 and 5 daphnids/vial). We found that particle aggregation occurs within the first 48 h of incubation in all treatments, no substantial change in the aggregate heterogeneity is observed afterwards. The median aggregate size was ∼2-fold higher than the nominal average particle size of clay and microplastics in the stock suspensions used to prepare the experimental mixtures. The strongest positive driver of the aggregate size and heterogeneity was DOM, followed by the presence of daphnids and the concentration of the suspended solids in the system. Also, microplastics were found to facilitate aggregation, albeit they were the weakest contributor. Moreover, besides directly increasing the aggregation, DOM relaxed the effects of the total solids and daphnids on the aggregate size. Thus, the particle size distribution was established early during the exposure and shaped by all experimental factors and their interactions. These findings improve our understanding of the processes occurring in the exposure systems when conducting effect studies with microplastics and other particulates and demonstrate the necessity to access the particle size distribution to characterise the exposure. Aslo, relevant experimental designs with microplastics must include relevant natural particulates and DOM to ensure environmentally realistic particle behaviour and adequate particle-biota interactions.

Simultaneous RNA and DNA Adductomics Using Single Data-Independent Acquisition Mass Spectrometry Analysis.

Adductomics studies are used for the detection and characterization of various chemical modifications (adducts) of nucleic acids and proteins. The advancements in liquid chromatography coupled with high-resolution tandem mass spectrometry (HRMS/MS) have resulted in efficient methods for qualitative and quantitative adductomics. We developed an HRMS-based method for the simultaneous analysis of RNA and DNA adducts in a single run and demonstrated its application using Baltic amphipods, useful sentinels of environmental disturbances, as test organisms. The novelty of this method is screening for RNA and DNA adducts by a single injection on an Orbitrap HRMS instrument using full scan and data-independent acquisition. The MS raw files were processed with an open-source program, nLossFinder, to identify and distinguish RNA and DNA adducts based on the characteristic neutral loss of ribonucleosides and 2'-deoxyribonucleosides, respectively. In the amphipods, in addition to the nearly 150 putative DNA adducts characterized earlier, we detected 60 putative RNA adducts. For the structural identification of the detected RNA adducts, the MODOMICS database was used. The identified RNA adducts included simple mono- and dimethylation and other larger functional groups on different ribonucleosides and deaminated product inosine. However, 54 of these RNA adducts are not yet structurally identified, and further work on their characterization may uncover new layers of information related to the transcriptome and help understand their biological significance. Considering the susceptibility of nucleic acids to environmental factors, including pollutants, the developed multi-adductomics methodology with further advancement has the potential to provide biomarkers for diagnostics of pollution effects in biota.

DNA Adductomics for the Biological Effect Assessment of Contaminant Exposure in Marine Sediments.

Exposure to chemical pollution can induce genetic and epigenetic alterations, developmental changes, and reproductive disorders, leading to population declines in polluted environments. These effects are triggered by chemical modifications of DNA nucleobases (DNA adducts) and epigenetic dysregulation. However, linking DNA adducts to the pollution load in situ remains challenging, and the lack of evidence-based DNA adductome response to pollution hampers the development and application of DNA adducts as biomarkers for environmental health assessment. Here, we provide the first evidence for pollution effects on the DNA modifications in wild populations of Baltic sentinel species, the amphipod Monoporeia affinis. A workflow based on high-resolution mass spectrometry to screen and characterize genomic DNA modifications was developed, and its applicability was demonstrated by profiling DNA modifications in the amphipods collected in areas with varying pollution loads. Then, the correlations between adducts and the contaminants level (polycyclic aromatic hydrocarbons (PAHs), trace metals, and pollution indices) in the sediments at the collection sites were evaluated. A total of 119 putative adducts were detected, and some (5-me-dC, N6-me-dA, 8-oxo-dG, and dI) were structurally characterized. The DNA adductome profiles, including epigenetic modifications, differed between the animals collected in areas with high and low contaminant levels. Furthermore, the correlations between the adducts and PAHs were similar across the congeners, indicating possible additive effects. Also, high-mass adducts had significantly more positive correlations with PAHs than low-mass adducts. By contrast, correlations between the DNA adducts and trace metals were stronger and more variable than for PAHs, indicating metal-specific effects. These associations between DNA adducts and environmental contaminants provide a new venue for characterizing genome-wide exposure effects in wild populations and apply DNA modifications in the effect-based assessment of chemical pollution.

High capacity for a dietary specialist consumer population to cope with increasing cyanobacterial blooms.

We present a common-garden experiment to examine the amphipod Monoporeia affinis, a key deposit-feeder in the Baltic Sea, a low diversity system offering a good model for studying local adaptations. In the northern part of this system, the seasonal development of phytoplankton is characterized by a single diatom bloom (high nutritional quality), whereas in the south, the diatom bloom is followed by a cyanobacteria bloom (low nutritional quality) during summer. Therefore, the nutrient input to the benthic system differs between the sea basins. Accordingly, the amphipod populations were expected to be dietary specialists in the north and generalists in the south. We tested this hypothesis using a combination of stable isotope tracers, trophic niche analyses, and various endpoints of growth and health status. We found that when mixed with diatomes, the toxin-producing cyanobacteria, were efficiently incorporated and used for growth by both populations. However, contrary to expectations, the feeding plasticity was more pronounced in the northern population, indicating genetically-based divergence and suggesting that these animals can develop ecological adaptations to the climate-induced northward cyanobacteria expansion in this system. These findings improve our understanding regarding possible adaptations of the deposit-feeders to increasing cyanobacteria under global warming world in both limnic and marine ecosystems. It is possible that the observed effects apply to other consumers facing altered food quality due to environmental changes.

Interspecific Interactions Drive Nonribosomal Peptide Production in Nodularia spumigena.

Nodularia spumigena is a bloom-forming cyanobacterium that produces several classes of nonribosomal peptides (NRPs) that are biologically active; however, the ecological roles of specific NRPs remain largely unknown. Here, we explored the involvement of NRPs produced by N. spumigena in interspecific interactions by coculturing the cyanobacterium and its algal competitors, the diatom Phaeodactylum tricornutum and the cryptomonad Rhodomonas salina, and measuring NRP levels and growth responses in all three species. Contrary to the expected growth suppression in the algae, it was N. spumigena that was adversely affected by the diatom, while the cryptomonad had no effect. Reciprocal effects of N. spumigena on the algae were manifested as the prolonged lag phase in R. salina and growth stimulation in P. tricornutum; however, these responses were largely attributed to elevated pH and not to specific NRPs. Nevertheless, the NRP levels in the cocultures were significantly higher than in the monocultures, with an up to 5-fold upregulation of cell-bound nodularins and exudation of nodularin and anabaenopeptin. Thus, chemically mediated interspecific interactions can promote NRP production and release by cyanobacteria, resulting in increased input of these compounds into the water. IMPORTANCE NRPs were involved in growth responses of both cyanobacteria and algae; however, the primary driver of the growth trajectories was high pH induced by N. spumigena. Thus, the pH-mediated inhibition of eukaryotic phytoplankton may be involved in the bloom formation of N. spumigena. We also report, for the first time, the reciprocal growth inhibition of N. spumigena by diatoms resistant to alkaline conditions. As all species in this study can co-occur in the Baltic Sea during summer, these findings are highly relevant for understanding ecological interactions in planktonic communities in this and other systems experiencing regular cyanobacteria blooms.

Microbiota-Dependent and -Independent Production of l-Dopa in the Gut of Daphnia magna.

Host-microbiome interactions are essential for the physiological and ecological performance of the host, yet these interactions are challenging to identify. Neurotransmitters are commonly implicated in these interactions, but we know very little about the mechanisms of their involvement, especially in invertebrates. Here, we report a peripheral catecholamine (CA) pathway involving the gut microbiome of the model species Daphnia magna. We demonstrate the following: (i) tyrosine hydroxylase and Dopa (3,4-dihydroxyphenylalanine) decarboxylase enzymes are present in the gut wall; (ii) Dopa decarboxylase gene is expressed in the gut by the host, and its expression follows the molt cycle peaking after ecdysis; (iii) biologically active l-Dopa, but not dopamine, is present in the gut lumen; (iv) gut bacteria produce l-Dopa in a concentration-dependent manner when provided l-tyrosine as a substrate. Impinging on gut bacteria involvement in host physiology and ecologically relevant traits, we suggest l-Dopa as a communication agent in the host-microbiome interactions in daphnids and, possibly, other crustaceans. IMPORTANCE Neurotransmitters are commonly implicated in host-microbiome communication, yet the molecular mechanisms of this communication remain largely elusive. We present novel evidence linking the gut microbiome to host development and growth via neurotransmitter l-Dopa in Daphnia, the established model species in ecology and evolution. We found that both Daphnia and its gut microbiome contribute to the synthesis of the l-Dopa in the gut. We also identified a peripheral pathway in the gut wall, with a molt stage-dependent dopamine synthesis, linking the gut microbiome to the daphnid development and growth. These findings suggest a central role of l-Dopa in the bidirectional communication between the animal host and its gut bacteria and translating into the ecologically important host traits suitable for subsequent testing of causality by experimental studies.

Understanding Biofilm Formation in Ecotoxicological Assays With Natural and Anthropogenic Particulates.

Fossil-made polymers harbor unique bacterial assemblages, and concerns have been raised that ingested microplastic may affect the consumer gut microbiota and spread pathogens in animal populations. We hypothesized that in an ecotoxicity assay with a mixture of polystyrene (PS) and clay: (1) microbiome of the test animals inoculates the system with bacteria; (2) relative contribution of PS and the total amount of suspended solids (SS) select for specific bacterial communities; and (3) particle aggregation is affected by biofilm community composition, with concomitant effects on the animal survival. Mixtures of PS and clay at different concentrations of SS (10, 100, and 1000 mg/L) with a varying microplastics contribution (%PS; 0-80%) were incubated with Daphnia magna, whose microbiome served as an inoculum for the biofilms during the exposure. After 4-days of exposure, we examined the biofilm communities by 16S rRNA gene sequencing, particle size distribution, and animal survival. The biofilm communities were significantly different from the Daphnia microbiota used to inoculate the system, with an overrepresentation of predatory, rare, and potentially pathogenic taxa in the biofilms. The biofilm diversity was stimulated by %PS and decreased by predatory bacteria. Particle aggregate size and the biofilm composition were the primary drivers of animal survival, with small particles and predatory bacteria associated with a higher death rate. Thus, in effect studies with solid waste materials, ecological interactions in the biofilm can affect particle aggregation and support potentially harmful microorganisms with concomitant effects on the test animals.

Competitive interactions as a mechanism for chemical diversity maintenance in Nodularia spumigena.

Nodularia spumigena is a bloom-forming diazotrophic cyanobacterium inhabiting brackish waters worldwide. This species produces non-ribosomal peptides (NRPs), including the hepatotoxin nodularin, often referred to as cyanotoxin. Several known classes of NRPs have various biological activities, although their modes of action are poorly understood. In the Baltic N. spumigena, there is a high NRP chemodiversity among strains, allowing their grouping in specific chemotypes and subgroups. Therefore, it is relevant to ask whether the NRP production is affected by intraspecific interactions between the co-existing strains. Using a novel approach that combines culture technique and liquid chromatography-tandem mass spectrometry for the NRP analysis, we examined N. spumigena strains under mono- and co-culture conditions. The test strains were selected to represent N. spumigena belonging to the same or different chemotype subgroups. In this setup, we observed physiological and metabolic responses in the test strains grown without cell contact. The changes in NRP levels to co-culture conditions were conserved within a chemotype subgroup but different between the subgroups. Our results suggest that intraspecific interactions may promote a chemical diversity in N. spumigena population, with higher NRP production compared to a single-strain population. Studying allelochemical signalling in this cyanobacterium is crucial for understanding toxicity mechanisms and plankton community interactions in the Baltic Sea and other aquatic systems experiencing regular blooms.

Ultraviolet Luminescence of ZnO Whiskers, Nanowalls, Multipods, and Ceramics as Potential Materials for Fast Scintillators.

The presented work is dedicated to the study and comparison of scintillating properties of zinc oxide samples prepared in different morphologies: whiskers, nanowalls, multipods, and ceramics. It was shown that total transmittance, photo- and radioluminescence spectra, and radioluminescence kinetics can vary significantly depending on sample structure and preparation conditions. The highest total transmittance was registered for ZnO ceramics (>50% at 0.5 mm thickness). Differences in the transmittance of whiskers, nanowalls, and multipods can be attributed to their shape and thickness which affects the amount of light refraction and scattering. The study of radioluminescence demonstrated that all samples, except undoped ceramics and air annealed whiskers, have predominantly fast luminescence with a decay time <1 ns. High transmittance of ceramics opens the way for their use in the registration of high energy X-ray and gamma radiation, where a large volume of scintillators is required. In cases, where large scintillator thickness is not a necessity, one may prefer to use other ZnO structures, such as ensembles of whiskers and nanowalls. Studies of near-band-edge luminescence components at low temperatures showed that the structure is quite similar in all samples except Ga doped ceramics.

Ecotoxicological assessment of suspended solids: The importance of biofilm and particle aggregation.

Assessment of microplastic impacts in biota is challenging due to the complex behavior of the test particles and their interactions with other particulates, including microorganisms, in the environment. To disentangle responses to microplastic exposure from those to other suspended solids, both microplastic and natural particles must be present in the test system. We evaluated how microplastic, non-plastic particles, and biofilms interacted in their effects on survivorship using acute toxicity assay with Daphnia magna. The animals were exposed to microplastic and kaolin at different concentrations of suspended solids (SS; 10, 100, and 1000 mg/L) with a varying microplastic contribution (%MP; 0-80%) and biofilm (presence/absence) associated with the solids. Also, we examined how these exposure parameters (SS, %MP, and Biofilm) affected aggregate formation that was analyzed using particle size distribution data. Under the exposure conditions, Daphnia mortality was primarily driven by SS concentration but ameliorated by both microplastic and biofilm. The ameliorating effects were related to increased particle aggregation in the presence of biofilm and high %MP. In addition, a weak yet significant positive effect of the biofilm on the survivorship was observed, presumably, due to microbial food supply to the daphniids in the exposure system; the bacteria were utilized at the absence of other food. Therefore, the effects of both natural and anthropogenic particulates depend on the particle behavior and aggregation in the water governed by microbial communities and physicochemical properties of the particles, which must be taken into account in the hazard assessment of plastic litter.

Antioxidant Responses in Copepods Are Driven Primarily by Food Intake, Not by Toxin-Producing Cyanobacteria in the Diet.

The association between oxidative processes and physiological responses has received much attention in ecotoxicity assessment. In the Baltic Sea, bloom-forming cyanobacterium Nodularia spumigena is a significant producer of various bioactive compounds, and both positive and adverse effects on grazers feeding in cyanobacteria blooms are reported. To elucidate the effect mechanisms and species sensitivity to the cyanobacteria-dominating diet, we exposed two Baltic copepods, Acartia bifilosa and Eurytemora affinis, to a diet consisting of toxin-producing cyanobacteria N. spumigena and a high-quality food Rhodomonas salina at 0-300 µg C L-1; the control food was R. salina provided as a monodiet at the same food levels. The subcellular responses to food type and availability were assayed using a suite of biomarkers - antioxidant enzymes [superoxide dismutases (SOD), catalase (CAT), and glutathione S-transferases (GST)] and acetylcholinesterase (AChE). In parallel, we measured feeding activity using gut content (GC) assayed by real-time PCR analysis that quantified amounts of the prey DNA in copepod stomachs. As growth and reproduction endpoints, individual RNA content (a proxy for protein synthesis capacity), egg production rate (EPR), and egg viability (EV%) were used. In both toxic and nontoxic foods, copepod GC, RNA content, and EPR increased with food availability. Antioxidant enzyme activities increased with food availability regardless of the diet type. Moreover, CAT (both copepods), SOD, and GST (A. bifilosa) were upregulated in the copepods receiving cyanobacteria; the response was detectable when adjusted for the feeding and/or growth responses. By contrast, the diet effects were not significant when food concentration was used as a co-variable. A bimodal response in AChE was observed in A. bifilosa feeding on cyanobacteria, with up to 52% increase at the lower levels (5-25 µg C L-1) and 32% inhibition at the highest food concentrations. These findings contribute to the refinement of biomarker use for assessing environmental stress and mechanistic understanding of cyanobacteria effects in grazers. They also suggest that antioxidant and AChE responses to feeding activity and diet should be accounted for when using biomarker profiles in field-collected animals in the Baltic Sea and, perhaps other systems, where toxic cyanobacteria are common.

Nitrogen isotope composition of amino acids reveals trophic partitioning in two sympatric amphipods.

According to ecological theory, two species cannot occupy the same niche. Using nitrogen isotope analyses (δ15N) of amino acids, we tested the extent to which two sympatric deposit-feeding amphipods, Monoporeia affinis and Pontoporeia femorata, partition their trophic resources. We found that trophic position (TP) and resynthesis index (∑V; a proxy for degradation status of ingested material prior to assimilation by the consumer) differ between species. The surface-feeding M. affinis had higher TP and intermediate ∑V, both pointing to a large contribution of metazoans in its diet. P. femorata, which feeds in the subsurface layers, had lower TP and a bimodal distribution of the ∑V values, supporting previous experimental evidence of a larger feeding niche. We also evaluated whether TP and ∑V values have consequences for amphipod fecundity and embryo viability and found that embryo viability in M. affinis was negatively linked to TP. Our results indicate that the amino acid-δ15N data paired with information about reproductive status are useful for detecting differences in the trophic ecology of sympatric amphipods.

Association between Aquatic Micropollutant Dissipation and River Sediment Bacterial Communities.

Assessment of micropollutant biodegradation is essential to determine the persistence of potentially hazardous chemicals in aquatic ecosystems. We studied the dissipation half-lives of 10 micropollutants in sediment-water incubations (based on the OECD 308 standard) with sediment from two European rivers sampled upstream and downstream of wastewater treatment plant (WWTP) discharge. Dissipation half-lives (DT50s) were highly variable between the tested compounds, ranging from 1.5 to 772 days. Sediment from one river sampled downstream from the WWTP showed the fastest dissipation of all micropollutants after sediment RNA normalization. By characterizing sediment bacteria using 16S rRNA sequences, bacterial community composition of a sediment was associated with its capacity for dissipating micropollutants. Bacterial amplicon sequence variants of the genera Ralstonia, Pseudomonas, Hyphomicrobium, and Novosphingobium, which are known degraders of contaminants, were significantly more abundant in the sediment incubations where fast dissipation was observed. Our study illuminates the limitations of the OECD 308 standard to account for variation of dissipation rates of micropollutants due to differences in bacterial community composition. This limitation is problematic particularly for those compounds with DT50s close to regulatory persistence criteria. Thus, it is essential to consider bacterial community composition as a source of variability in regulatory biodegradation and persistence assessments.

Calmodulin inhibition as a mode of action of antifungal imidazole pharmaceuticals in non-target organisms.

To improve assessment of risks associated with pharmaceutical contamination of the environment, it is crucial to understand effects and mode of action of drugs in non-target species. The evidence is accumulating that species with well-conserved drug targets are prone to be at risk when exposed to pharmaceuticals. An interesting group of pharmaceuticals released into the environment is imidazoles, antifungal agents with inhibition of ergosterol synthesis as a primary mode of action in fungi. However, imidazoles have also been identified as competitive antagonists of calmodulin (CaM), a calcium-binding protein with phylogenetically conserved structure and function. Therefore, imidazoles would act as CaM inhibitors in various organisms, including those with limited capacity to synthesize sterols, such as arthropods. We hypothesized that effects observed in crustaceans exposed to imidazoles are related to the CaM inhibition and CaM-dependent nitric oxide (NO) synthesis. To test this hypothesis, we measured (i) CaM levels and its gene expression, (ii) NO accumulation and (iii) gene expression of NO synthase (NOS1 and NOS2), in the cladoceran Daphnia magna exposed to miconazole, a model imidazole drug. Whereas significantly increased CaM gene expression and its cellular allocation were observed, supporting the hypothesized mode of action, no changes occurred in either NO synthase expression or NO levels in the exposed animals. These findings suggest that CaM inhibition by miconazole leads to protein overexpression that compensates for the loss in the protein activity, with no measurable downstream effects on NO pathways. The inhibition of CaM in D. magna may have implications for effect assessment of exposure to mixtures of imidazoles in aquatic non-target species.

Rapid Physicochemical Changes in Microplastic Induced by Biofilm Formation.

Risk assessment of microplastic (MP) pollution requires understanding biodegradation processes and related changes in polymer properties. In the environment, there are two-way interactions between the MP properties and biofilm communities: (i) microorganisms may prefer some surfaces, and (ii) MP surface properties change during the colonization and weathering. In a 2-week experiment, we studied these interactions using three model plastic beads (polyethylene [PE], polypropylene [PP], and polystyrene [PS]) exposed to ambient bacterioplankton assemblage from the Baltic Sea; the control beads were exposed to bacteria-free water. For each polymer, the physicochemical properties (compression, crystallinity, surface chemistry, hydrophobicity, and surface topography) were compared before and after exposure under controlled laboratory conditions. Furthermore, we characterized the bacterial communities on the MP surfaces using 16S rRNA gene sequencing and correlated community diversity to the physicochemical properties of the MP. Significant changes in PE crystallinity, PP stiffness, and PS maximum compression were observed as a result of exposure to bacteria. Moreover, there were significant correlations between bacterial diversity and some physicochemical characteristics (crystallinity, stiffness, and surface roughness). These changes coincided with variation in the relative abundance of unique OTUs, mostly related to the PE samples having significantly higher contribution of Sphingobium, Novosphingobium, and uncultured Planctomycetaceae compared to the other test materials, whereas PP and PS samples had significantly higher abundance of Sphingobacteriales and Alphaproteobacteria, indicating possible involvement of these taxa in the initial biodegradation steps. Our findings demonstrate measurable signs of MP weathering under short-term exposure to environmentally relevant microbial communities at conditions resembling those in the water column. A systematic approach for the characterization of the biodegrading capacity in different systems will improve the risk assessment of plastic litter in aquatic environments.

Mercury-methylating bacteria are associated with copepods: A proof-of-principle survey in the Baltic Sea.

Methylmercury (MeHg) is a potent neurotoxin that biomagnifies in marine food webs. Inorganic mercury (Hg) methylation is conducted by heterotrophic bacteria inhabiting sediment or settling detritus, but endogenous methylation by the gut microbiome of animals in the lower food webs is another possible source. We examined the occurrence of the bacterial gene (hgcA), required for Hg methylation, in the guts of dominant zooplankters in the Northern Baltic Sea. A qPCR assay targeting the hgcA sequence in three main clades (Deltaproteobacteria, Firmicutes and Archaea) was used in the field-collected specimens of copepods (Acartia bifilosa, Eurytemora affinis, Pseudocalanus acuspes and Limnocalanus macrurus) and cladocerans (Bosmina coregoni maritima and Cercopagis pengoi). All copepods were found to carry hgcA genes in their gut microbiome, whereas no amplification was recorded in the cladocerans. In the copepods, hgcA genes belonging to only Deltaproteobacteria and Firmicutes were detected. These findings suggest a possibility that endogenous Hg methylation occurs in zooplankton and may contribute to seasonal, spatial and vertical MeHg variability in the water column and food webs. Additional molecular and metagenomics studies are needed to identify bacteria carrying hgcA genes and improve their quantification in microbiota.