Fungi are well associated with the degradation of hydrocarbons by the production of different enzymes, among which catalases (CBH), laccases (LCC) and peroxidases (LiP and MnP) are of immense importance. In this study, crude oil tolerance and enzyme secretions were demonstrated by rhizospheric fungal strains. Four most abundant strains were isolated from the rhizosphere of grasses growing in aged oil spill sites and identified through morphological characterization and molecular PCR-amplification of 5.8–28S ribosomal rRNA using ITS1 and ITS4 primers. These strains were subjected to crude oil tolerance test at 0–20% concentrations. Presence and transcriptase responses of putative genes lig (1–6), mnp, cbh (1.1, 1.1 and 11), and lcc encoding lignin peroxidase, manganese peroxidase, catalase, and laccase enzymes respectively were also studied in these strains using RT-PCR. In addition, activities of secreted enzymes by each strain were studied in aliquots. The strains were identified as Aspergillus niger asemoA (KY473958), Talaromyces purpurogenus asemoF (KY488463), Trichoderma harzianum asemoJ (KY488466), and Aspergillus flavus asemoM (KY488467) through sequencing and comparing the sequences’ data at NCBI BLAST search software. All the isolated strains showed tolerance to crude oil at 20% concentration, but the growth rate reduced with increasing in oil concentrations. All the isolated strains possess the tested genes and lig 1–6 gene was overexpressed in A. niger and T. harzianum while lcc and mnp genes were moderately expressed in all the four strains. Almost 145 U.mL⁻¹ of lignin and manganese peroxidase, 87 U.mL⁻¹ of catalase, and 180 U.mL⁻¹ of laccase enzymes were produced by these strains and it was also observed that these strain mostly produced studied enzymes in response to increasing crude oil concentrations. Considering the robust nature and diverse production of these catalytic enzymes by these strains, they can be exploited for various bioremediation technologies as well as other biotechnological applications.

Copper nanoparticles (nCu) are widely used in industry and in daily life, due to their unique physical, chemical, and biological properties. Few studies have focused on nCu phytotoxicity, especially with regard to toxicity mechanisms in crop plants. The present study examined the effect of 15.6 μM nCu exposure on the root morphology, physiology, and gene transcription levels of wheat (Triticum aestivum L.), a major crop cultivated worldwide. The results obtained were compared with the effects of exposing wheat to an equivalent molar concentration of ionic Cu (Cu²⁺ released from CuSO₄) and to control plants. The relative growth rate of roots decreased to approximately 60% and the formation of lateral roots was stimulated under nCu exposure, possibly due to the enhancement of nitrogen uptake and accumulation of auxin in lateral roots. The expression of four of the genes involved in the positive regulation of cell proliferation and negative regulation of programmed cell death decreased to 50% in the Cu²⁺ treatment compared to that of the control, while only one gene was down-regulated to about half of the control in nCu treatment. This explained the decreased root cell proliferation and higher extent of induced cell death in Cu²⁺- than in nCu-exposed plants. The increased methane dicarboxylic aldehyde accumulation (2.17-fold increase compared with the control) and decreased antioxidant enzyme activities (more than 50% decrease compared with the control) observed in the Cu²⁺ treatment in relation to the nCu treatment indicated higher oxidative stress in Cu²⁺- than in nCu-exposed plants. Antioxidant (e.g., proline) synthesis was pronouncedly induced by nCu to scavenge excess reactive oxygen species, alleviating phytotoxicity to wheat exposed to this form of Cu. Overall, oxidative stress and root growth inhibition were the main causes of nCu toxicity.

Glyphosate (GLY) is one of the most used herbicide worldwide. Considering that information concerning the impact of GLY on bivalves is scarce, in this study we evaluated for the first time the effects of environmentally realistic concentrations of GLY (10, 100 and 1000 μg/L) to the mussel Mytilus galloprovincialis. Mussels were exposed for 7, 14 and 21 days and several biomarkers were measured in haemocytes/haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (antioxidant enzyme and acetylcholinesterase activities). The concentrations of GLY and its main metabolite aminomethylphosphonic acid in the experimental tanks were also measured. The MANOVA analysis demonstrated that the experimental variables considered (exposure concentration, exposure duration, and their interaction) affected significantly biomarker responses. In addition, the two-way ANOVA analysis indicated that GLY was able to affect most of the cellular parameters measured, whereas antioxidant enzyme activities resulted to be influenced moderately. Interestingly, exposure to GLY reduced significantly acetylcholinesterase activity in gills. Although preliminary, the results of this study demonstrated that GLY can affect both cellular and biochemical parameters in mussels, highlighting a potential risk for aquatic invertebrates.

The gulf of Annaba, the most important touristic and economic coastal zone located in Northeast Algeria, is contaminated by several pollutants from urban, agricultural, harbor and industrial activities. Elevated levels of heavy metals were detected in a locally prevalent edible mollusk Donax trunculus (Bivalvia, Donacidae) widely used as a sentinel species for the assessment of marine pollution. The present work aims to measure the difference between two localities, one being full of different pollutants (Sidi Salem) and the other being relatively clean (El Battah) and to evaluate the ability of D. trunculus to overcome the environmental stress during a transplantation experiment by a determination of fatty acid profile, the enzymes activities and the level of metallothioneins (MTs), a biomarker of metallic contamination. Adults of D. trunculus were collected at Sidi Salem (contaminated site) and transplanted into El Battah (reference site) for 21 days in cages (60 × 60 × 60 cm with a 2 mm mesh). Biochemical analyzes were conducted at different times (0, 7, 14 and 21 days). At 0-day experiment: the rate of the fatty acids, the enzymes activities and MT levels at the site of Sidi Salem (polluted site) were significantly different from those of El Battah. During the transplantation a gradual restoration of fatty acids rates, enzymes activities and MT levels was observed. At the end of the period of transplantation, the values are comparable to those of El Battah. A two-way ANOVA (time, site) on data revealed significant effects of time and site. Overally, D. trunculus is able to induce its detoxification system and to restore relatively rapidly the status of individuals from the reference site (El Battah).

Herein, we utilize sequential extraction and high-throughput sequencing to investigate the effects of nanomaterial additives on As volatilization from flooded soils. We reveal that maximum volatilization is achieved in the fourth week and is followed by stabilization. The extent of volatilization decreased in the order of control > nano-zerovalent iron >40-nm hydroxyapatite > nano-Fe₃O₄ > 20-nm hydroxyapatite > multilayer graphene oxide > high-quality graphene oxide. The most abundant forms of As in soil corresponded to As-Fe and Al oxides. In soil with low levels of As pollution, the contents of these species increased after treatment with graphene oxides but decreased after treatment with other nanomaterials, with an opposite trend observed for soil with high levels of As pollution. The addition of nanomaterials influenced the activity of soil enzymes, e.g., hydroxyapatites affected the activities of urease and alkaline phosphatase, whereas graphene oxides significantly impacted that of peroxidase (P < 0.05). The addition of nanomaterials (which can potentially inhibit microbial growth) affected As levels by influencing the amount of As volatilized from polluted soil. Moreover, As volatilization, enzyme activity, and As speciation were observed to be mutually correlated (e.g., volatilization was negatively correlated to peroxidase activity and the contents of amorphous crystalline hydrous oxides of As-Fe and Al).

The occurrence and transportation of antibiotics in biofilms from natural and engineered sources have attracted increasing interests. Nevertheless, the effects of extracellular polymeric substances (EPS) on the responses of biofilms to the exposure to antibiotics are not clear. In this study, the effects of EPS on the sorption and biological responses to one representative antibiotic, sulfamethizole (STZ), in model biofilms were investigated. Proteins dominated the interactions between the EPS and the STZ and the EPS from a moving bed biofilm reactor exhibited the strongest interaction with the STZ. The EPS served as important reservoirs for the STZ and the tested biofilms all showed reduced sorption capacities for the STZ after the EPS were extracted. The respiratory rates and typical enzymatic activities were reduced after the EPS were extracted. High-throughput 16S rRNA gene sequencing results confirmed that the bacterial community in the biofilm without the EPS was more vulnerable to antibiotic shock as indicated by the community diversity and richness indices. A greater increase in the abundance of susceptible species was observed in the natural biofilm. The results comprehensively suggested that the EPS played important role in biosorption of STZ and alleviated the direct damage of the antibiotic to the cells; in addition the extent of the bacterial community response was associated with the origins of the biofilms. Our study provided details on the responses of multi-species biofilms to the exposure to an antibiotic and highlighted the role of the EPS in interacting with the antibiotic, thereby providing a deeper understanding of the bioremediation of antibiotics in real-life natural and engineered biofilm systems.

Heavy metal tolerance of microorganisms is the basis of heavy metal removal by growing cells. In this study, a cross-protection effect generated by salt stress significantly enhanced the cadmium tolerance of multi-stress-tolerant Pichia kudriavzevii. Comparative transcriptome analysis using RNA-Seq linked with physiological and biochemical observation was used to elucidate the underlying mechanisms of the improved cadmium tolerance. The expression of cadmium transport related genes (GSTY2, GLR1, GLO2, YCF1 and YOR1), GSH content and GST activity were elevated by salt stress, suggesting enhanced cadmium conjugation and detoxification in yeast cells. The inhibited cadmium uptake by ZRT1 and enhanced cadmium efflux by YOR1 contributed to the decrease in the intracellular cadmium concentration. The improved expression of antioxidant enzyme genes (SOD1, SOD2, SOD6, CAT1 and PRXIID), along with the enhanced activities of antioxidant enzymes (SOD, CAT and POD) resulted in a decrease in cadmium-induced ROS production, protein carbonylation, lipid peroxidation and cell death. The abundant expression of heat shock protein genes (HSP12, HSP10 and SSC1) and genes related to trehalose synthesis (TPS1 and TSL1) induced by salt stress protected yeast cells against complex stress conditions, contributing to the improved cadmium tolerance. These findings will be useful to develop cadmium-tolerant yeasts for cadmium removal by growing cells.

Microcystins (MCs) are naturally occurring algal toxins in the aquatic environment and pose a serious threat to the ecosystem. In general, aquatic populations are structured by organisms of different ages, with varying degrees of biochemical and physiological responses. In this study, juvenile and adult marine mysids (Neomysis awatschensis) were exposed to MC-Leucine Arginine (MC-LR) (0.1, 1, and 10 μg L⁻¹) for 7 days, and the bioconcentration dynamics and responses of antioxidant defense system were measured during the exposure and additional depuration periods (7 days). MC-LR bioconcentrated in a dose-dependent manner, from a threshold concentration of 1 μg L⁻¹ in both stages, and the levels reduced gradually during the depuration phase. Bioconcentration patterns of MC-LR were highly age-specific, as juvenile mysids showed peaks during the exposure period, whereas adults exhibited a peak on the first day of depuration. After exposure to 10 μg L⁻¹ concentration, elevated levels of malondialdehyde (MDA) and glutathione (GSH) were observed during the late (days 5 and 7) exposure and early (days 1 and 3) depuration periods in juvenile mysids, while adult mysids showed a peak on day 7 of the exposure period. Age-specific responses were also observed in the enzymatic activities of glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR). Juvenile mysids showed a significant elevation in all enzymatic activities during the exposure and/or depuration phase upon exposure to 10 μg L⁻¹ MC-LR, but only CAT and SOD enzymes showed significant changes during the exposure and/or depuration periods in adults. Overall, our results indicate the bioconcentration potential of MC-LR and its threshold in the marine mysid, in addition to age-specific MC-LR dynamics and subsequent biochemical responses.

A mobile pilot plant was set up in a wastewater treatment plant (WWTP) in southwest Spain to address potential adverse effects of effluents as a whole contaminant, which are discharging into marine environments. Ruditapes philippinarum specimens were exposed to different effluent concentrations (50%, 25%, 12.5%, 6.25%, and 3.15%) during seven days. After effluent exposure, lysosomal membrane stability alterations (LMS), changes in the energy status storage (total lipids content (TLP) and in the mitochondrial electron transport (MET), inhibition of inflammatory mechanisms (cyclooxygenase activity (COX)), and neurotoxic effects (acetylcholinesterase (AChE) were determined in exposed organisms. Furthermore, potential toxic reduction in the effluent was analysed by the application of an additional microalgae tertiary treatment called photobiotreatment (PhtBio). Results after PhtBio confirmed the toxic effect reduction in exposed organisms. Neuroendocrine effects, alterations in energy budget and in lipid storage revealed alterations in clam's health status causing stress conditions after effluent exposure.

Soil algal bioassays have been limited by their inability to evaluate several toxic endpoints because it is difficult to collect pure soil algae growing on and beneath the soil surface. This study describes the extension of a previously developed paper-disc method for analyzing soil toxicity to algae. The method can be used in conjunction with flow cytometric analysis and facilitates the assessment of previously proposed toxicity endpoints, such as the growth zone, biomass, and photosynthetic activity. We assessed the applicability of this paper-disc soil method using the green algae Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata exposed to nickel-contaminated soil; examined cell sizes, cell granularity, enzyme activity, and oxidative stress as new toxicity endpoints using flow cytometry; and identified morphological changes in green algae assayed. The results showed that, used in conjunction with flow cytometry, the extended paper-disc soil method is sufficiently sensitive to detect decreases in cell granularity in C. reinhardtii and esterase activity in P. subcapitata. The method also revealed decreases in growth zone, biomass, and electron transfer from the reaction center to the quinone pool. Collectively, the results of this study indicate that soil algal bioassays using nonspecific algae can be used to assess soil quality, to derive several toxicity endpoints for individual cells, and to evaluate previously established flow cytometric toxicity endpoints.