This study investigates the metabolism and mode of action of galaxolide (HHCB) in the European sea bass -Dicentrarchus labrax- following a single intraperitoneal injection of 50 mg HHCB/kg body weight. In addition, a group of fish was injected with 50 mg/kg of ketoconazole (KCZ), a fungicide that is known to interfere with different Cyp isoenzymes. HHCB was actively metabolised by sea bass and acted as a weak inhibitor of the synthesis of oxyandrogens in gonads of male fish. Both, HHCB and a hydroxylated metabolite were detected in bile. The fungicide ketoconazole was a strong inhibitor of Cyp11β and Cyp3a-catalyzed activities. The work contributes to the better understanding of the impact of synthetic musks on fish and proposes the determination of HHCB and/or its hydroxylated metabolite in bile as a tool to assess environmental exposure in wild fish.

Effects of canopy ozone (O3) exposure and signatures of genetic structure using isozyme markers associated with O3 tolerance were analyzed in 20-, 80-, and >200-yr-old ponderosa (Pinus ponderosa Dougl. ex Laws.) and Jeffrey pine (Pinus jeffreyi Grev. & Balf.) in Sequoia National Park, California. For both species, the number of alleles and genotypes per loci was higher in parental trees relative to saplings. In ponderosa pine, the heterozygosity value increased, and the fixation index indicated reduction of homozygosity with increasing tree age class. The opposite tendencies were observed for Jeffrey pine. Utilizing canopy attributes known to be responsive to O3 exposure, ponderosa pine was more symptomatic than Jeffrey pine, and saplings were more symptomatic than old growth trees. We suggest that these trends are related to differing sensitivity of the two species to O3 exposure, and to higher O3 exposures and drought stress that younger trees may have experienced during germination and establishment. Genetic variation in isozyme markers associated with ozone tolerance differed between parental trees and their progeny in two closely related species of yellow pine.

Rice is a staple crop, and food chain contamination of arsenic in rice grain possesses a serious health risk to billions of population. Arsenic stress negatively affects the rice growth, yield and quality of the grains. Nitric oxide (NO) is a major signaling molecule that may trigger various cellular responses in plants. The protective role of NO during arsenite (AsIII) stress and its relationship with plant physiological and metabolic responses is not explored in detail. Exogenous NO, supplemented through the roots in the form of sodium nitroprusside, has been shown to provide protection vis-à-vis AsIII toxicity. The NO-mediated variation in physiological traits such as stomatal density, size, chlorophyll content and photosynthetic rate maintained the growth of the rice plant during AsIII stress. Besides, NO exposure also enhanced the lignin content in the root, decreased total arsenic content and maintained the activities of antioxidant isoenzymes to reduce the ROS level essential for protecting from AsIII mediated oxidative damage in rice plants. Further, NO supplementation enhanced the GSH/GSSG ratio and PC/As molar ratio by modulating PC content to reduce arsenic toxicity. Further, NO-mediated modulation of the level of GA, IAA, SA, JA, amino acids and phenolic metabolites during AsIII stress appears to play a central role to cope up with AsIII toxicity. The study highlighted the role of NO in AsIII stress tolerance involving modulation of metalloid detoxification and physiological pathways in rice plants.

Cadmium (Cd) pollution in mangrove wetlands has received increasing attention as urbanization expands rapidly. As a dominant mangrove species, Kandelia obovata is highly tolerant to Cd toxicity. Plant hormones and superoxide dismutase (SODs) play critical roles in the response to heavy metal stress in K. obovata roots. Although theirs important influence have been reported, the regulation mechanism between SODs and plant hormones in Cd detoxification by K. obovata roots remains limited. Here, we investigated relationships among SOD, plant hormones, and Cd tolerance in K. obovata roots exposed to Cd. We found that Cd was retained in the epidermis and exodermis of roots, and the epidermis and exodermis had highest hydrogen peroxide (H2O2) content and SOD activity. Similarly, SOD isozymes also exhibited distinct activity in the different parts of root. Overexpressed KoCSD3 and KoFSD2 individually in Nicotiana benthamiana revealed that different SOD members contributed to H2O2 content regulation by promote the activity of downstream antioxidant enzymes under Cd treatment. In addition, assays on the effects of hormones showed that increased endogenous indole-3-acetic acid (IAA) was observed in the cortex and stele, whereas the abscisic acid (ABA) content was enhanced in the epidermis and exodermis in roots during Cd treatment. The results of exogenous hormones treatment indicated that KoFSD2 upregulated under ABA and IAA treatment, but KoCSD3 only induced by ABA stimulation. Taken together, our results reveal the relationship between SODs and plant hormones, which expands the knowledge base regarding KoSODs response to plant hormones and mediating H2O2 concentration under Cd stress.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants and could produce oxidative toxicity to plants. Our previous study has shown that miR398 is involved in response to phenanthrene treatment by targeting CSD1 and CSD2. However, it is not clear which is essential for CSD1 and CSD2 and how miR398 changes. In this study, we performed discontinuous PAGE to separate superoxide dismutase (SOD) isozymes and found that two bands of the cytosolic Cu/Zn-SOD are induced by phenanthrene at day 5 and 7. Low expression of pri-miR398 and high expression of pre-miR398 indicate that the conversion process from pri-miR398 to pre-miR398 is impeded, which causes decrease in mature miR398. The relative expression of CSD1 is entirely up-regulated, further confirming the important role of CSD1 in response to phenanthrene exposure. Besides, the overexpression of WRKY implies its potential function in answering the call from phenanthrene stress. Therefore, it is concluded that the gene silencing of CSD1 recedes due to the biosynthesis inhibition of miR398, causing the increase of SOD activity in response to phenanthrene exposure in wheat roots. Our results are useful not only for better understanding miRNAs regulation in detoxication of reactive oxygen species, but also for alleviating the toxicity to crops caused by PAHs.

Acetylcholinesterase (AChE) is a reliable biomarker of pesticide exposure although in clams this activity is often very low or undetectable. Carboxylesterases (CEs) exhort several physiological roles, but also respond to pesticides. Searching for an AChE alternative, baseline CE activities were characterised in Ruditapes decussatus gills and digestive glands using five substrates suggestive of different isozymes. The long chain p-nitrophenyl butyrate and 1-naphthyl butyrate were the most sensitive. In the digestive gland, their kinetic parameters (Vmax and Km) and in vitro sensitivity to the organophosphorus metabolite chlorpyrifos oxon (CPX) were calculated. IC50 values, in the pM–nM range, suggest a high protection efficiency of CE-related enzymes towards CPX neurotoxicity. Other targeted enzymes were: activities of glutathione reductase, glutathione peroxidase, catalase, glutathione S-transferases (GSTs) and lactate dehydrogenase in gills and digestive glands. The high GSTs activity and CE/AChE ratio suggests that R. decussatus has a great capacity for enduring pesticide exposure.

Aquaculture activities are often established in the vicinity of highly populated, potentially contaminated areas. Animals cultured at such locations, namely bivalves, are frequently used as test organisms in ecotoxicological testing. In this case, a period of depuration is required to allow the normalization of physiological processes, which are likely to be altered after exposure to a multiplicity of waterborne contaminants occurring in the wild. One of the most important species in modern marine aquaculture is the oyster species Crassostrea gigas. The aim of this study was to assess if the current depuration time frame of 24 h (adopted by most aquaculture facilities), is long enough to permit oysters to revert potential toxic effects exerted by environmental contaminants, allowing their use in laboratory-based ecotoxicological studies. The selected approach involved the monitoring of biochemical (antioxidant defence, oxidative damage, phase II metabolism, and neurological homeostasis) and physiological (condition index) parameters, along a period of 42 days. The obtained results showed that a period of 24 h does not revert any of the potential toxic effects caused by environmental contaminants to which animals may have been previously subjected; even a period of 42 days was not long enough for the oysters to completely normalize the levels of their antioxidant defences, namely total GPx activity, which increased over time. Lipid peroxidation was also increased during the depuration period, and the activity of the metabolic isoenzymes GSTs was significantly decreased. Furthermore, AChE activity measured in the adductor muscle of oysters was increased over time. These assumptions suggest that a period of depuration longer than 24 h is mandatory to obtain adequate test organisms of this oyster species, to be used for ecotoxicological testing purposes.

Benzo[a]pyrene (B[a]P) is an environmental toxicant and endocrine disruptor. Therefore, the aim of the present study was to investigate the toxicity of B[a]P in testis of rats and also to study the role of silymarin and thymoquinone (TQ) as natural antioxidants in the alleviation of such toxicity. Data of the present study showed that levels of testosterone, estrogen and progesterone were significantly decreased after treatment of rats with B[a]P. In addition, B[a]P caused downregulation of the expressions of steroidogenic enzymes including CYP17A1 and CP19A1, and decreased the activity of 17-β hydroxysteroid dehydrogenase (17β-HSD). Moreover, B[a]P decreased the activities of antioxidant enzymes including catalase (CAT), glutathione peroxidase (GPX) and superoxide dismutase (SOD), and significantly increased free radicals levels in testis of male rats. However, pretreatment of rats with silymarin prior to administration of B[a]P was found to restore the level of free radicals, antioxidant status, and activities of steroidogenic enzymes to their normal levels in testicular tissues. Moreover, histopathological finding showed that silymarin recovered the abnormalities occurred in tubules caused by B[a] P in testis of rats. On the other hand, TQ showed pro-oxidant effects and did not ameliorate the toxic effects of B[a] P on the testicular tissue since it decreased antioxidant enzymes activities and inhibited the protein expression of CYP11A1 and CYP21A2 compared to control rats. Moreover, TQ decreased the levels of testosterone, estrogen, and progesterone either in the presence or absence of B[a]P. It is concluded that B[a]P decreased testosterone levels, inhibited antioxidant enzymes activities, caused downregulation of CYP isozymes involved in steroidogenesis, and increased free radical levels in testis. Moreover, silymarin was more effective than TQ in restoring organism health and alleviating the deleterious effects caused by B[a]P in the testis of rats. Due to its negative impact, it is highly recommended to limit the use of TQ as a dietary supplement since millions of people in the Middle East are using it to improve their health.

Hospital wastewater (HWW) contains different hazardous substances resulting from a combination of medical and non-medical activities of hospitals, including pharmaceutical residues. These substances may represent a threat to the aquatic environment if they do not follow specific treatment processes. Therefore, we aimed to investigate the effects of the untreated effluent collected from a general hospital in Mahdia City (Tunisia) on neonatal stages of the freshwater crustacean Daphnia magna. Test organisms were exposed to three proportions (3.12%, 6.25%, and 12.5% v/v) of HWW. After 48 h of exposure, a battery of biomarkers was measured, including the quantification of antioxidant enzymes [catalase (CAT) and total and selenium-dependent glutathione peroxidase (total GPx; Se-GPx)], phase II biotransformation isoenzymes glutathione-S-transferases (GSTs), cyclooxygenases (COX) involved in the regulation of the inflammatory process, and total cholinesterases (ChEs) activities. Lipid peroxidation (LPO) was measured to estimate oxidative damage. The here-obtained results showed significant decreases of CAT and GSTs activities and also on LPO content in daphnids, whereas Se-GPx activity was significantly increased in a dose-dependent manner. Impairment of cholinesterasic and COX activities were also observed, with a significant decrease of ChEs and an increase of COX enzymatic activities. Considering these findings, HWW was capable of inducing an imbalance of the antioxidant defense system, but without resulting in oxidative damage in test organisms, suggesting that peroxidases and alternative detoxifying pathways were able to prevent the oxidant potential of several drugs, which were found in the tested effluents. In general, this study demonstrated the toxicity of hospital effluents, measured in terms of the potential impairment of key pathways, namely neurotransmission, antioxidant defense, and inflammatory homeostasis of crustaceans.

This paper examined the gut digestive enzymes, trypsin and leucine aminopeptidase (LAP), in gypsy moth (Lymantria dispar, L.) larvae exposed to cadmium. We analyzed the 3-day acute effect, chronic effects from hatching until sacrifice, and recovery from long-term dietary treatment with cadmium concentrations of 10 and 30 μg Cd/g dry food. The activities of both examined enzymes declined at the higher level of cadmium after both acute and chronic treatments and did not recover within 3 days of feeding a diet with no added cadmium. Leucine aminopeptidase was more sensitive because its activity was inhibited after both short-term treatments. Three trypsin and one leucine aminopeptidase isoform were detected by electrophoresis. Egg hatches (full-sib families) differed in enzyme activities, index of phenotypic plasticity, and isozyme expression after different treatments. Statistically significant positive correlations between these enzymes pointed to common genetic regulation. Moreover, variances were higher for the control group than for cadmium treatment groups implying that these proteolytic enzymes did not participate directly in detoxification. These results suggest that, with additional research to discover the mechanisms of enzyme inhibition, trypsin and leucine aminopeptidase might be used as biomarkers to indicate the severity of gastrointestinal disease due to cadmium intake.