Due to the large-scale outbreak of Corona Virus Disease (2019), amounts of disinfecting agents was regularly used in public environments and their potential toxicity towards organisms needed to be appreciated. Thus, one mostly used cationic disinfectant, benzalkonium chlorides (BAC(C12)), was selected to assess its potential toxicity one common cyanobacteria Microcystis aeruginosa (M. aeruginosa) in this study. The aims were to explore the toxic effect and mechanism of BAC (C12) on M. aeruginosa growth within 96 h via morphological, physiological, and the relative and absolute quantification (iTRAQ)-based quantitative proteomics variations. The results found that BAC(C12) significantly inhibited cell density of M. aeruginosa at concentrations from 1 mg/L to 10 mg/L, and the 96-h EC₅₀ value was identified to be 3.61 mg/L. Under EC₅₀ concentration, BAC(C12) depressed the photosynthesis activities of M. aeruginosa exhibited by 36% decline of the maximum quantum yield for primary photochemistry (Fv/Fm) value and denaturation of photosynthetic organelle, caused oxidative stress response displayed by the increase of three indexes including superoxide dismutase (SOD), malondialdehyde (MDA), and the intracellular reactive oxygen species (ROS), and destroyed the integrity of cell membranes demonstrated by TEM images and the increase of ex-cellular substances. Then, the iTRAQ-based proteomic analysis demonstrated that BAC(C12) depressed photosynthesis activities through inhibiting the expressions of photosynthetic protein and photosynthetic electron transport related proteins. The suppression of electron transport also led to the increase of superoxide radicals and then posed oxidative stress on cell. Meantime, the 63.63% ascent of extracellular microcystin production of M. aeruginosa was observed, attributing to the high expression of microcystin synthesis proteins and the damage of cell membrane. In sum, BAC(C12) exposure inhibited the growth of M. aeruginosa mainly by depressing photosynthesis, inducing oxidative stress, and breaking the cell membrane. And, it enhanced the release of microcystin from the cyanobacterial cells via up-regulating the microcystin synthesis proteins and inducing the membrane damage, which could enlarge its toxicity to aquatic species.

Systemic studies on the bioremediation of co-contaminated PAHs and heavy metals are lacking, and this paper provides an in-depth review on the topic. The released sources and transport of co-contaminated PAHs and heavy metals, including their co-occurrence through formation of cation-π interactions and their adsorption in soil are examined. Moreover, it is investigated that co-contamination of PAHs and heavy metals can drive a synergistic positive influence on bioremediation through enhanced secretion of extracellular polymeric substances (EPSs), production of biosynthetic genes, organic acid and enzymatic proliferation. However, PAHs molecular structure, PAHs-heavy metals bioavailability and their interactive cytotoxic effects on microorganisms can exert a challenging influence on the bioremediation under co-contaminated conditions. The fluctuations in bioavailability for microorganisms are associated with soil properties, chemical coordinative interactions, and biological activities under the co-contaminated PAHs-heavy metals conditions. The interactive cytotoxicity caused by the emergence of co-contaminants includes microbial cell disruption, denaturation of DNA and protein structure, and deregulation of antioxidant biological molecules. Finally, this paper presents the emerging strategies to overcome the bioavailability problems and recommends the use of biostimulation and bioaugmentation along with the microbial immobilization for enhanced bioremediation of PAHs-heavy metals co-contaminated sites. Better knowledge of the bioremediation potential is imperative to improve the use of these approaches for the sustainable and cost-effective remediation of PAHs and heavy metals co-contamination in the near future.

In this study, the contents of microcystin-LR (MC-LR) of Microcystis aeruginosa cultures in the laboratory and natural water samples from the Huangpu River in different seasons were detected through enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC), respectively. Excellent correlation between the two methods was obtained (R² > 0.99). ELISA was a reliable and simple method with high reproducibility (coefficient of variation < 25%) and satisfactory recovery for the monitoring of low levels of MC-LR. MC-LR concentrations in Huangpu River varied with the seasonal variation, which peaked in August with the temperature over 30 °C and then gradually declined with the decreasing temperature after August. The highest MC-LR concentration in the Huangpu River was below the WHO drinking water quality standard (1 µg/L). These results indicated that warm temperature accelerated the MC-LR synthesis and release, and it is necessary to regularly monitor the MC-LR levels, especially during the high algae period in summer. ELISA can be applied to detect the low levels of MC-LR in the field without complex treatment, avoiding the samples from denaturation and degradation during the transportation. Hence, ELISA is a better alternative of HPLC when HPLC is unavailable, especially when rapid testing is required in routine MC-LR analysis.

Collagen is a promising candidate for food and pharmaceutical applications due to its excellent biocompatibility, low antigenicity, and controlled biodegradability; however, its heavy price restricts its utilization. Fish scales generated during the processing are generally regarded as waste material and an environmental pollutant, though they are a promising source of collagen. In the present study, Cirrhinus mrigala scales were demineralized and extracted for acid-soluble collagen (ASC) using acetic acid, with a collagen yield of 2.7%. UV–Vis spectra, SDS-PAGE, FTIR analyses, and amino acid composition confirmed the type I nature of the collagen extracted. The denaturation temperature of the collagen was found to be 30.09 °C using differential scanning calorimetry (DSC). The collagen was highly soluble at acidic pH and lower NaCl concentrations while its solubility was lowered in alkaline conditions and NaCl concentrations above 0.5 M. The collagen exhibited good emulsifying potential with an emulsion activity index (EAI) and emulsion stability index (ESI) of 21.49 ± 0.22 m² g⁻¹ and 15.67 ± 0.13 min, respectively. Owing to the good physicochemical characteristics of the extracted collagen, collagen-chitosan-neem extract (CCN) films were prepared subsequently which showed good antimicrobial activity against Bacillus subtilis NCIM 2635, Staphylococcus aureus NCIM 2654, Escherichia coli NCIM 2832, and Pseudomonas aeruginosa NCIM 5032, suggesting the potential of collagen in the development of antimicrobial films. These results demonstrate that the collagen from fish waste could be valorized and used effectively along with chitosan and neem extract for the synthesis of novel biodegradable films with antimicrobial efficacy.

The collagens (COL2, COL4, and COL5) were extracted from chrome-tanned leather shavings via three distinctive routes of acid method. The dechroming degree of COL2 extracted with the easiest operation was the highest (95.6% ± 1.2%) and the yield exceeded 90%; however, the total amount of acid was the most and the cost was the highest. In the second route, although the three-step dechroming process brought cumbersome operation, the dechroming degree and yield of COL4 were 90.5% ± 0.8% and 92.2% ± 0.6%, respectively, and the acid amount was less than that in the first route. For COL5, the dechroming degree and yield was the lowest; nevertheless, this route had the advantages of lowest cost and simpler operation. Electrophoretic patterns showed that all the collagens contained α1, α2, and β chains without low molecular weight components and were close to those of type I collagen. Compared with native collagen extracted from fresh calf skin, the regenerated collagens also maintained unique triple helix conformation determined via ultraviolet, infrared spectra and X-ray diffraction, confirmed by the similar values of AIII/A₁₄₅₅ and Δν. Additionally, the collagens existed in the form of fibrils with D-period pattern of ~ 67 nm. Furthermore, the denaturation temperatures of COL2, COL4, and COL5 were 71.2, 79.1, and 85.4 °C, respectively, which were relevant to the tighter arrangement of fibrils with the increased chromium content.

As an alternative to bisphenol A, bisphenol S (BPS) is widely used in industrial production and daily life, which is then discharged into sewage treatment plants and accumulates in sludge. In this research, impact and interaction mechanism of BPS on lipids hydrolysis in sludge is studied from the respect of soluble organic matter and volatile organic fatty acids (VFAs). Multi-spectra, thermodynamics, molecule docking, and enzyme activity assay are applied to elucidate the effect mechanism of BPS on lipids hydrolysis. Results show that lipids hydrolysis is restrained due to the denaturation of lipase with BPS exposure. The interaction mechanism is involved in hydrophobic bond and hydrogen bond interaction in the activity region of lipase. This interaction not only results in an unfolding skeleton structure of lipase and a less hydrophobic microenvironment of tyrosine and tryptophan residues but also leads to fluorophore static quenching with the formation of lipase-BPS complex. The experimental results and the combined research methods not only contribute to the development of novel technique for sludge treatment containing micropollutant but also profit to clarify the interaction mechanism between other micropollutant and enzymes.

The aim of this study was to adapt the Comet assay in spermatozoa of the marine prawn Palaemon serratus to use it as a marker of sperm quality. Indeed, due to the characteristics of their spermatozoa, the measurement of DNA integrity is one of the few markers which can be transferred to crustaceans to assess the quality of their semen. In the first step, the methods of collecting and maintaining spermatozoa were optimized. Cell survival was estimated during kinetics of preservation (i.e. 1, 2, 4 and 8 h) in various suspension media to define artificial seawater (ASW) as optimal. Several methods in the releasing of spermatozoa from the spermatophore of prawns were estimated with regard to their incidence both on the efficiency of extraction and the survival of cells. Pipetting up and down turned out to be the most successful and the least invasive technique. Secondly, the transfer of Comet assay was optimized by studying various times in both cell lysis (i.e. 1, 6, 18 h) and DNA denaturation (i.e. 15, 30 and 45 min), after in vitro exposure of spermatozoa to an H2O2 gradient as model genotoxicant. Results revealed that a minimum of 1 h in cell lysis and 15 min of DNA denaturation were sufficient to obtain valuable results, linked with a low compaction of DNA in spermatozoa of Palaemon sp. Finally, the sensitivity of P. serratus spermatozoa was assessed after in vitro exposures to model genotoxicants displaying various modes of interaction with DNA (i.e. UV-C, 13.3–79.5 J m⁻²; H₂O₂, 5–10 μM and MMS, 0.5–5 mM) and some environmental contaminants known or suspected to be genotoxic (i.e. cadmium and diuron, 0.015–1.5 μg L⁻¹; carbamazepine, 0.1–10 μg L⁻¹) for invertebrates. The low variability of the baseline level of DNA strand breaks recorded in controls highlighted the robustness of the method. P. serratus spermatozoa displayed significant DNA damage from the lowest doses tested for all model genotoxicants, but conversely, no genotoxic effect of tested environmental contaminants was observed. These results, which are discussed according to the protocol tested in the present study and the comparison with literature data, could suggest a difference in the response or sensitivity of spermatozoa to environmental genotoxicity between invertebrate species, and therefore the interest of Palaemonidae prawns in ecogenotoxicology. In conclusion, the present study underlines the potential of the Comet assay as a marker to assess the contamination impact on the sperm quality in Palaemonidae prawns in view to a potential application for in situ biomonitoring surveys.

Increases in global wildfires and fire severity are expected to result from global warming. Severe wildfires not only burn surface vegetation but also affect forest soil. Humic substances play key roles in the transport of nutrients and the carbon cycle in terrestrial ecosystems. In this study, we evaluated the effects of forest fires on the chemical properties of fulvic acid (FA) and humic acid (HA) extracted from non-burned and burned forest soils in Gunma, Japan. The differential thermal analysis of FA indicated that the intensity of exothermic reaction peak at 400 °C was 2-fold higher than that from non-burned soil. Based on pyrolysis-gas chromatography-mass spectrometry analysis with tetramethyl ammonium hydroxide, the amount of pyrolysate compounds in FA from burnt soil was significantly lower than that in FA from non-burnt soil. Therefore, we can conclude that the forest fire caused the significant change in the properties of FA such as increasing the aromaticity and refractory. In addition, the concentration of dissolved organic carbon with low molecular weight in surface soil increased after forest fire. This study suggests that the denaturation of soil organic matter by wildfire can affect the carbon cycle in terrestrial ecosystems.

An increasing amount of heavy metals (e.g., Cu²⁺) is being discharged into sewage treatment plants and is accumulating in sludge, which is toxic to the enzyme in sludge or soil when the sludge is used as fertilizer, resulting in unfavorable effect on the biological treatment of sludge and the circulation and conversion of materials in soil. In this research, effect of Cu²⁺ on sludge hydrolysis by α-amylase is studied from the respect of concentration and components of soluble organic matter in sludge, using three-dimensional fluorescence spectra. Results show that Cu²⁺ exposure not only inhibits the hydrolysis of sludge due to the denaturation of α-amylase but also affects the components of soluble organic matter in sludge. In order to illuminate the interaction mechanism between Cu²⁺ and α-amylase (a model of hydrolase in sludge), multi-spectra and isothermal titration microcalorimetry techniques are applied. Results show that the secondary structure of α-amylase is changed as that the α-helical content increases and the structure loosens. The microenvironment of amino acid residue in α-amylase is changed that the hydrophobicity decreases and the polarity increases with Cu²⁺ exposure. Isothermal titration calorimetry results show that Van der Waals force and hydrogen bond exist in the interaction between Cu²⁺ and α-amylase. Results from this research would favor the development of advanced process for the biological treatment of sludge containing heavy metals.

To better understand the mechanisms of TiO₂ nanoparticle (NP) uptake and toxicity in aquatic organisms, we investigated the interaction of NPs with the proteins found in gill mucus from blue mussels. Mucus is secreted by many aquatic organisms and is often their first line of defense against pathogens, xenobiotics, and other sources of environmental stress. Here, five TiO₂ NPs and one SiO₂ NP were incubated with gill mucus and run out on a one-dimensional polyacrylamide gel for a comparative qualitative analysis of the free proteins in the mucosal solution and the proteins bound to NPs. We then used nanoscale liquid chromatography coupled with tandem mass spectrometry to identify proteins of interest. Our data demonstrated dissimilar protein profiles between the crude mucosal solution and proteins adsorbed on NPs. In particular, extrapallial protein (EP), one of the most abundant mucus proteins, was absent from the adsorbed proteins. After thermal denaturation experiments, this absence was attributed to the EP content in aromatic amino acids that prevents protein unfolding and thus adsorption on the NP. Moreover, although the majority of the protein corona was qualitatively similar across the NPs tested here (SiO₂ and TiO₂), a few proteins in the corona showed a specific recruitment pattern according to the NP oxide (TiO₂ vs SiO₂) or crystal structure (anatase TiO₂ vs rutile TiO₂). Therefore, protein adsorption may vary with the type of NP. Graphical abstract Proteins with adsorption selectivity as identified from isolated bands