Metals and metalloids are natural components of the biosphere, which are not produced per se by human beings, but whose form and distribution can be affected by human activities. Like all substances, they are a contaminant if present in excess compared to background levels and/or in a form that would not normally occur in the environment. Samples of liver, gills, gonads and muscle from European chub, Squalius cephalus, were analyzed for Al, As, B, Ba, Cr, Cu, Fe, Hg, Mn, Mo, Sr and Zn using inductively coupled plasma optical emission spectrometry (ICP-OES) to highlight the importance of tissue selection in monitoring research. The comet assay or single cell gel electrophoresis (SCGE) was selected as an in vivo genotoxicity assay, a rapid and sensitive method for measuring genotoxic effects in blood, liver and gills of the European chub. Microscopic images of comets were scored using Comet IV Computer Software (Perceptive Instruments, UK).The objective of our study was to investigate two reservoirs, Zlatar and Garasi, and one river, Pestan by: (i) determining and comparing metal and metalloid concentrations in sediment, water and tissues of European chub: liver, gills, muscle and gonads (ii) comparing these findings with genotoxicity of water expressed through DNA damage of fish tissues.A clear link between the level of metals in water, sediment and tissues and between metal and genotoxicity levels at examined sites was not found. This suggests that other xenobiotics (possibly the organic compounds), contribute to DNA damage.

Arsenic exposure contributed to the development of human diseases. Arsenic exerted multiple organ toxicities mainly by triggering oxidative stress. However, the signaling pathway underlying oxidative stress is unclear. We previously found that the expression of SFPQ, a splicing factor, was positively associated with urinary arsenic concentration in an arsenic-exposed population, suggesting an oxidative stress regulatory role for SFPQ. To test this hypothesis, we established cell models of oxidative stress in human hepatocyte cells (L02) treated with NaAsO₂. Reactive oxygen species (ROS) synthesis displayed a time- and dose-dependent increase with NaAsO₂ treatment. SFPQ suppression resulted in a 36%–53% decrease in ROS generation, leading to enhanced cellular damage determined by 8-OHdG, comet tail moment, and micronucleus analysis. Particularly, SFPQ deficiency attenuated expression of the oxidase genes DUOX1, DUOX2, NCF2, and NOX2. A fluorescent-based RNA electrophoretic mobility shift assay (FREMSA) and dual-luciferase reporter system revealed that miR-92b-5p targeted DUOX2 mRNA degradation. An RNA immunoprecipitation assay showed an interaction between SFPQ and miR-92b-5p of the miRNA-induced silencing complex (miRISC). Notably, NaAsO₂ treatment diminished the interaction between SFPQ and miR92b-5p, accompanied by decreased binding between miR-92b-5p and 3′-UTR of DUOX2. However, SFPQ deficiency suppressed the dissociation of miR-92b-5p from 3′-UTR of DUOX2, indicating that miR-92b-5p regulated the SFPQ-dependent DUOX2 expression. Taken together, we reveal that SFPQ responds to arsenic-induced oxidative stress by interacting with the miRISC. These findings offer new insight into the potential role of SFPQ in regulating cellular stress response.

A heterotrophic arsenite [As(III)]-oxidizing bacterium Agrobacterium tumefaciens GW4 isolated from As(III)-rich groundwater sediment showed high As(III) resistance and could oxidize As(III) to As(V). The As(III) oxidation could generate energy and enhance growth, and AioR was the regulator for As(III) oxidase. To determine the related metabolic pathways mediated by As(III) oxidation and whether AioR regulated other cellular responses to As(III), isobaric tags for relative and absolute quantitation (iTRAQ) was performed in four treatments, GW4 (+AsIII)/GW4 (-AsIII), GW4-ΔaioR (+AsIII)/GW4-ΔaioR (-AsIII), GW4-ΔaioR (-AsIII)/GW4 (-AsIII) and GW4-ΔaioR (+AsIII)/GW4 (+AsIII). A total of 41, 71, 82 and 168 differentially expressed proteins were identified, respectively. Using electrophoretic mobility shift assay (EMSA) and qRT-PCR, 12 genes/operons were found to interact with AioR. These results indicate that As(III) oxidation alters several cellular processes related to arsenite, such as As resistance (ars operon), phosphate (Pi) metabolism (pst/pho system), TCA cycle, cell wall/membrane, amino acid metabolism and motility/chemotaxis. In the wild type with As(III), TCA cycle flow is perturbed, and As(III) oxidation and fermentation are the main energy resources. However, when strain GW4-ΔaioR lost the ability of As(III) oxidation, the TCA cycle is the main way to generate energy. A regulatory cellular network controlled by AioR is constructed and shows that AioR is the main regulator for As(III) oxidation, besides, several other functions related to As(III) are regulated by AioR in parallel.

This study aims to evaluate toxic effects of exposure to chlortetracycline (CTC) in soil on reproductive endpoints (juvenile counts and cocoon counts), biochemical responses, and genotoxic potentials of the earthworm Eisenia fetida. Results showed that juvenile counts and cocoon counts of the tested earthworms were reduced after exposure to CTC. The effective concentrations (EC₅₀ values) for juvenile and cocoon counts were 96.1 and 120.3 mg/kg, respectively. Treatment of earthworms with CTC significantly changed the activity of catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST). An increase in malondialdehyde (MDA) indicated that CTC could cause cellular lipid peroxidation in the tested earthworms. The percentage of DNA in the tail of single-cell gel electrophoresis of coelomocytes as an indication of DNA damage increased after treatment with different doses of CTC, and a dose-dependent DNA damage of coelomocytes was found. In conclusion, CTC induces physiological responses and genotoxicity on earthworms.

Bioaugmentation, Biostimulation and Horizontal gene transfer (HGT) of catabolic genes have been proven for their role in bioremediation of hydrocarbons. It also has been proved that selection of either biostimulation or bioremediation varies for every contaminated site. The reliability of HGT compared to biostimulation and bioremediation was not tested. The present study focuses on reliability of biostimulatiion, bioaugmentation and HGT during biodegradation of Diesel oil and Non aqueous phase liquids (NAPL). Pseudomonas aeruginosa (AEBBITS1) having alkB and NDO genes was used for bioaugmentation and the experiment was conducted using seawater as medium. Based on Gas chromatography results diesel was found to be degraded to 100% in both presence and absence of AEBBITS1. Denturing gradient gel electrophoresis result showed same pattern in presence and absence of AEBBITS1 indicating no HGT. NAPL degradation was found to be more by Biostimulated Bioaugmentation compared to biostimulation and bioaugmentation alone. This proves that biostimulated bioaugmentation is better strategy for oil contamination (tarabll) in Velsao beach, Goa.

Vitellogenin (Vtg) is a well-established biomarker of estrogenic exposure in aquatic animals. In vertebrates, Vtg gene transcription is controlled by the estrogen receptors (ERs). Although an ER ortholog is present in molluscs, its role as a transcriptional regulator remains elusive. Here, we tested the hypothesis that in the Sydney rock oyster, Saccostrea glomerata, the ER ortholog activates Vtg gene transcription through specific interaction with its promoter. Luciferase reporter assays indicated that sgER activated both a minimal promoter containing the consensus estrogen-responsive elements (EREs) and the sgVtg promoter in an estrogen-independent manner. The sgVtg promoter-luciferase activation was significantly reduced when any of three putative ERE half sites (½EREs) in the promoter were mutated. Electrophoretic mobility shift assay (EMSA) confirmed that sgER binds specifically to a 68-bp promoter sequence where these ½EREs reside. Overall, the results suggest that sgER is a constitutively active transcription factor that binds and activates the sgVtg promoter.

Genotoxicity of 5-fluorouracil (5-FU), etoposide (ET) and cadmium chloride (CdCl₂) was evaluated in Limnodrilus udekemianus, cosmopolitan tubificid species, by alkaline single-cell gel electrophoresis (comet assay). Groups of 50 individuals were exposed in vivo in water-only short-term (96 h) tests to 5-FU (0.004, 0.04, 0.4, 4 and 40 μM), ET (0.004. 0.04, 0.4 and 4 μM) and CdCl₂ (0.004, 0.04, 0.4, 4 and 40 μM). Mortality of worms was observed only for CdCl₂ (4 and 40 μM). Cell viability lower than 70 % was detected for 5-FU (0.4, 4 and 40 μM), ET (4 μM) and CdCl₂ (0.4 and 4 μM). All tested substances induced significant increase of DNA damage except 0.004 μM of ET. L. udekemianus being sensitive to all tested substances indicates that it can be used in ecogenotoxicology studies. Concern should be raised to cytostatics, especially to 5-FU, since concentration of 0.004 μM induced DNA damage is similar to ones detected in wastewaters.

The gill tissue is the main site of metabolic enzymes or compensation, with the kidney tissue playing a supporting role. At the gill tissue, carbonic anhydrase enzymes (CAs) catalyze the hydration of CO₂ to HCO₃⁻ and H⁺ for production to the H₂O. In this work, the CA enzyme was purified from horse mackerel (Trachurus trachurus) gill with a specific activity of 21,381.42 EU/mg, purification fold of 150.61, total activity of 2347.68 EU/mL, and a yield of 16.13% using sepharose 4B-L-tyrosine-sulfanilamide affinity gel chromatography. For recording the enzyme purity, gel electrophoresis was performed, and single band was seen. The molecular weight of this enzyme was found approximately 35 kDa. Also, the inhibitory effects of different pesticides such as thiram, clofentezine, propineb, deltamethrin, azoxystrobin, and thiophanate and heavy metal ions such as Fe²⁺, Cu²⁺, Co²⁺, Pb²⁺ Hg²⁺, and As³⁺ on horse mackerel gill tissue CA enzyme activities were investigated. Our results indicated that these pesticides and metal ions showed inhibitory effects at low nanomolar and millimolar concentrations for fish gill CA enzymes, respectively.

The effects of irrigation with meat processing factory effluent (MPE) in combination with additions of paunch to three arable sites and one pasture site on soil chemical and microbial properties were investigated in fields surrounding a beef meat processing factory. A pasture site that had only received MPE was also sampled along with adjoining arable and pasture control fields that had never received MPE or paunch. Additions of MPE/paunch caused increases in electrical conductivity, exchangeable Na and K, exchangeable sodium percentage (ESP), extractable P, organic C, total N, microbial biomass C, and metabolic quotient and decreases in exchangeable Ca and Mg, pH, and the proportion of organic C present as microbial biomass. The structure and diversity of bacterial and fungal communities was measured by polymerase chain reaction–denaturing gradient gel electrophoresis of 16S rDNA and internal transcribed spacer-RNA amplicons respectively and catabolic diversity by analysis of catabolic response profiles to 25 substrates. Principal component analysis of catabolic response profiles clearly separated control from MPE/paunch-treated sites, and this was associated with greater catabolic responses to the carboxylic acids α-ketoglutaric, α-ketobutyric, L-ascorbic, and citric acid in the control. At the arable sites, application of MPE and paunch caused increases in bacterial, fungal, and catabolic diversity. Canonical correspondence analysis of the relationship between catabolic, bacterial, and fungal fingerprints and soil properties indicated that the main soil variables separating MPE/paunch treatments from controls were the higher organic C, ESP, and extractable P and a lower pH, exchangeable Ca, and Mg. It was concluded that, although long-term MPE/paunch additions induce soil salinity, sodicity, and acidity, in general, they cause an increase in the size, activity, and structural and functional diversity of in the soil microbial community.

Imazalil (IMZ), a fungicide containing imidazole group, is extensively used for the prevention and treatment of fungal diseases in plants. Current study was performed to examine cyto-genotoxic potential of IMZ on Allium cepa roots by following Allium ana-telophase and single cell gel electrophoresis (comet) assays. The concentration which reduced the growth of the root tips of IMZ by 50% compared to the negative control group (EC₅₀) was found to be 1 μg/mL by Allium root growth inhibition test. 0.5, 1, and 2 μg/mL concentrations of IMZ were exposed to Allium roots for intervals of 24, 48, 72, and 96 h. 10 μg/mL of methyl methane sulfonate (MMS) and distilled water were used as control groups, both positive and negative. Statistical analysis was performed by using one-way ANOVA with Duncan’s multiple comparison tests at p ≤ 0.05 and Pearson correlation test at p = 0.01. IMZ showed cytotoxic effect by statistically decreasing root growth and mitotic index (MI) and also genotoxic effect by statistically increasing chromosomal aberrations (CAs) and DNA damage compared to the negative control group. With these cyto-genotoxic effects, it should be used carefully and further cyto-genotoxic mechanisms should be investigated along with other toxicity tests.