Silver nanomaterials (AgNMs) are broadly used and among the most studied nanomaterials. The underlying molecular mechanisms (e.g. protein and metabolite response) that precede phenotypical effects have been assessed to a much lesser extent. In this paper, we assess differentially expressed proteins (DEPs) and metabolites (DEMs) by high-throughput (HTP) techniques (HPLC-MS/MS with tandem mass tags, reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC) with mass spectrometric detection). In a time series (0, 7, 14 days), the standard soil model Enchytraeus crypticus was exposed to AgNM300K and AgNO₃ at the reproduction EC20 and EC50. The impact on proteins/metabolites was clearly larger after 14 days. NM300K caused more upregulated DEPs/DEMs, more so at the EC20, whereas AgNO₃ caused a dose response increase of DEPs/DEMs. Similar pathways were activated, although often via opposite regulation (up vs down) of DEPs, hence, dissimilar mechanisms underlie the apical observed impact. Affected pathways included e.g. energy and lipid metabolism and oxidative stress. Uniquely affected by AgNO₃ was catalase, malate dehydrogenase and ATP-citrate synthase, and heat shock proteins (HSP70) and ferritin were affected by AgNM300K. The gene expression-based data in Adverse Outcome Pathway was confirmed and additional key events added, e.g. regulation of catalase and heat shock proteins were confirmed to be included. Finally, we observed (as we have seen before) that lower concentration of the NM caused higher biological impact. Data was deposited to ProteomeXchange, identifier PXD024444.

The present study investigated the effects of dietary iron (Fe) exposure on physiological performance and homeostatic regulation of trace metals during development (5–28 days post-fertilization; dpf) in zebrafish (Danio rerio). The results demonstrated that whole body Fe content was increased in 14 dpf larvae fed a high Fe diet. Cumulative mortality was also significantly elevated during exposure to the high Fe diet. Using droplet digital PCR, we observed that high Fe-exposed larvae exhibited an increase in mRNA levels of the Fe-storage protein ferritin, which appeared to be associated with the elevated level of whole body Fe content. Further, the results indicated that dietary Fe exposure induced transient changes in the mRNA expression levels of various metal transporters, including the iron transporter dmt1, and the zinc transporters zip8 and zip14. The expression of the epithelial Ca²⁺ channels (i.e., ecac) was also found to increase by high dietary Fe. Overall, our findings suggest that larval fish during the early nutritional transition period are sensitive to the effects of dietary Fe.

Intersex, the appearance of female characteristics in male gonads, has been identified in several aquatic species. It is a widespread phenomenon in populations of the bivalve, Scrobicularia plana, from the southwest coast of the U.K. Genes previously identified as differentially expressed (ferritin, testicular haploid expressed gene, THEG, proliferating cell nuclear antigen, PCNA; receptor activated protein kinase C, RACK; cytochrome B, CYB; and cytochrome c oxidase 1, COX1) in intersex clams relative to normal male clams, were selected for characterisation and an environmental survey of the Channel region. Transcripts were significantly differentially expressed at sites with varying intersex incidence and contaminant burdens. Significant correlations between specific gene expressions, key contaminants and sampling locations have been identified, though no single gene was associated with intersex incidence. The results highlight the difficulty in understanding the intersex phenomenon in molluscs where there is still a lack of knowledge on the control of normal reproduction.

Hepatic gene expression and liver histology were examined in sand flathead (Platycephalus bassensis) from six locations in Port Phillip Bay, Victoria, Australia. Four sets of genes including thyroid-related genes (D1, D2, TTR, TRα and TRβ), metal metabolism-related genes (MT, MTF1, TF, Ferritin and FPN1), apoptosis-related genes (Diablo/SMAC1, Diablo/SMAC2 and CYP1A) and an endoplasmic reticulum stress biomarker gene (GRP78) were examined in female flathead using qRT-PCR. TRβ and Diablo/SMAC1 gene expression was significantly up-regulated in fish from all polluted sites compared to those from a reference site. The transcripts of TRα and FPN1 were significantly higher in flathead from Corio Bay, while the hepatic mRNA of TTR and GRP78 were significantly lower in those fish. Positive correlations were observed between Diablo/SMAC1 and CYP1A, D2 and TRβ, TRα and TRβ. This study demonstrates that application of pathway-based biomarker genes and histopathology can provide comprehensive information on the impact of environmental pollutants on fish.

The metal pollution has posed great risk on the coastal organisms along the Jiulongjiang Estuary in South China. In this work, two-dimensional electrophoresis-based proteomics was applied to the oysters Crassostrea hongkongensis from metal pollution sites to characterize the proteomic responses to metal pollution. Metal accumulation and proteomic responses indicated that the oysters from BJ site were more severely contaminated than those from FG site. Compared with those oyster samples from the clean site (JZ), metal pollution induced cellular injuries, oxidative and immune stresses in oyster heapatopancreas from both BJ and FG sites via differential metabolic pathways. In addition, metal pollution in BJ site induced disturbance in energy and lipid metabolisms in oysters. Results indicated that cathepsin L and ferritin GF1 might be the biomarkers of As and Fe in oyster C. hongkongensis, respectively. This study demonstrates that proteomics is a useful tool for investigating biological effects induced by metal pollution.

In this study, the adsorption kinetic of cesium, strontium, and rubidium radionuclides was investigated using ferritin magnetic molecules. Kinetic investigation of synthetic and natural wastes was carried out and the results were compared. Pseudo first-order, pseudo second-order, Elovich, double-exponential, and intraparticle diffusion models were the kinetic models used in the fitting of experimental data. The kinetic study of synthetic waste revealed that the double-exponential model demonstrated excellent fitting. Coefficient of determination resulting from fitting of cesium, strontium, and rubidium radionuclide’s adsorption results via the double-exponential model are 0.9938, 0.9905, and 0.9863, respectively. In the experiments conducted on natural wastes, too, all of the five kinetic models were investigated. Results indicated that the double-exponential model matched greatly with the experimental data, and cesium, strontium, and rubidium radionuclide’s coefficients of determination were 0.9742, 0.9613, and 0.9442, respectively. Comparison of the results of natural and synthetic wastes showed that matching with the model and recovery of target elements were more prominent in experiments with synthetic waste (unicomponent) rather than natural waste (multicomponent).

Calcareous soil, high pH, and low organic matter are the major factors that limit iron (Fe) availability to rice crop. The present study was planned with the aim to biofortified rice grain with Fe, by integrated use of chemical and organic amendments in pH-manipulated calcareous soil. The soil pH was reduced (pHL₂) by using elemental sulfur (S) at the rate of 0.25 % (w/w). The organic amendments, biochar (BC) and poultry manure (PM) [1 % (w/w)], along with ferrous sulfate at the rate of 7.5 mg kg⁻¹ soil were used. The incorporation of Fe with BC in soil at pHL₂ significantly improved plant biomass, photosynthetic rate, and paddy yield up to 99, 97, and 36 %, respectively, compared to control. A significant increase in grain Fe (190 %), protein (58 %), and ferritin (400 %) contents was observed while anti-nutrients, i.e., polyphenols (37 %) and phytate (21 %) were significantly decreased by the addition of Fe and BC in soil at pHL₂ relative to control. Among the organic amendments, PM significantly increased Cd, Pb, Ni, and Cr concentrations in rice grain relative to control but their concentration values were below as compared to the toxic limits of hazard quotients and hazard index (HQ and HI). Hence, this study implies that Fe applied with BC in the soil at pHL₂ can be considered as an effective strategy to augment Fe bioavailability and to reduce non-essential heavy metal accumulation in rice grain.

Krill oil (KO) is a recent supplement which is rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). These fatty acids are found in both krill oil and fish oil. In krill oil, they esterified to phospholipids, but in fish oil, they are esterified to triacylglycerols. The target of this study was to investigate whether KO could help against iron overload–induced toxicity in liver and spleen. Rats were randomly assigned into 3 categories: control rats, rats received iron in a drinking water for 8 weeks followed by either vehicle or KO (40 mg/kg) treatment for an extra 8 weeks. Extent of hepatic and splenic injury was assessed via biochemical, histopathological and immunohistochemical evaluations. KO effectively improved the microscopic features of liver and spleen. Moreover, it decreased the increased levels of serum transaminases, ALP, LDH, iron, and ferritin and increased albumin serum level as well. In addition, it restored the balance between oxidants and antioxidants in the hepatic and splenic tissues. Furthermore, it decreased HO-1 levels, upregulated the production of Nrf2, and limited the expression of MMP9. These findings altogether suggest that KO might be a new candidate for treatment of iron overload-induced toxicity. Graphical abstract Graphical abstract

Rice (Oryza sativa L.) is one of the main staple food crops which is inherently low in micronutrients, especially iron (Fe), and can lead to severe Fe deficiency in populations having higher consumption of rice. Soils polluted with nickel (Ni) can cause toxicity to rice and decreased Fe uptake by rice plants. We investigated the potential role of biochar (BC) and gravel sludge (GS), alone and in combination, for in situ immobilization of Ni in an industrially Ni-contaminated soil at original and sulfur-amended altered soil pH. Our further aim was to increase Fe bioavailability to rice plants by the exogenous application of ferrous sulfate to the Ni-immobilized soil. Application of the mixture of both amendments reduced grain Ni concentration, phytate, Phytate/Fe, Phyt/Zn molar ratios, and soil DTPA-extractable Ni. In addition, the amendment mixture increased 70 % Fe and 229 % ferritin concentrations in rice grains grown in the soil at original pH. The Fe and ferritin concentrations in S-treated soil was increased up to 113 and 383 % relative to control respectively. This enhanced Fe concentration and corresponding ferritin in rice grains can be attributed to Ni/Fe antagonism where Ni has been immobilized by GS and BC mixture. This proposed technique can be used to enhance growth, yield, and Fe biofortification in rice by reducing soil pH while in parallel in situ immobilizing Ni in polluted soil.

Association of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements with cytosolic proteins of different molecular masses was described for the liver of European chub (Squalius cephalus) from weakly contaminated Sutla River in Croatia. The principal aim was to establish basic trace element distributions among protein fractions characteristic for the fish living in the conditions of low metal exposure in the water. The fractionation of chub hepatic cytosols was carried out by size exclusion high performance liquid chromatography (SE-HPLC; Superdex™ 200 10/300 GL column), and measurements were performed by high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Elution profiles of essential elements were mostly characterized by broad peaks covering wide range of molecular masses, as a sign of incorporation of essential elements in various proteins within hepatic cytosol. Exceptions were Cu and Fe, with elution profiles characterized by sharp, narrow peaks indicating their probable association with specific proteins, metallothionein (MT), and ferritin, respectively. The main feature of the elution profile of nonessential metal Cd was also single sharp, narrow peak, coinciding with MT elution time, and indicating almost complete Cd detoxification by MT under the conditions of weak metal exposure in the water (dissolved Cd concentration ≤0.3 μg L⁻¹). Contrary, nonessential metal Pb was observed to bind to wide spectrum of proteins, mostly of medium molecular masses (30–100 kDa), after exposure to dissolved Pb concentration of ~1 μg L⁻¹. The obtained information within this study presents the starting point for identification and characterization of specific metal/metalloid-binding proteins in chub hepatic cytosol, which could be further used as markers of metal/metalloid exposure or effect on fish.