Heavy metals uptake by the global economic crop (Pisum sativum L.) grown in contaminated soils and its associated health risks.

The aim of the present investigation was to determine the concentration of heavy metals in the different organs of Pisum sativum L. (garden pea) grown in contaminated soils in comparison to nonpolluted soils in the South Cairo and Giza provinces, Egypt, and their effect on consumers' health. To collect soil and plant samples from two nonpolluted and two polluted farms, five quadrats, each of 1 m2, were collected per each farm and used for growth measurement and chemical analysis. The daily intake of metals (DIM) and its associated health risks (health risk index (HRI) were also assessed. The investigated heavy metals were cadmium (Cd), arsenic (As), chromium (Cr), copper (Cu), nickel (Ni), iron (Fe), manganese (Mn), zinc (Zn), silver (Ag), cobalt (Co) and vanadium (V). Significant differences in soil heavy metals, except As, between nonpolluted and polluted sites were recorded. Fresh and dry phytomass, photosynthetic pigments, fruit production, and organic and inorganic nutrients were reduced in the polluted sites, where there was a high concentration of heavy metals in the fruit. The bioaccumulation factor for all studied heavy metals exceeded 1 in the polluted sites and only Pb, Cu and Mn exceeded 1 in the nonpolluted sites. Except for Fe, the DIM of the studied heavy metals in both sites did not exceed 1 in either children or adults. However, the HRI of Pb, Cd, Fe, and Mn in the polluted plants and Pb in the nonpolluted ones exceeded 1, indicating significant potential health risks to consumers. The authors recommend not to eat garden peas grown in the polluted sites, and farmers should carefully grow heavy metals non-accumulating food crops or non-edible plants for other purposes such as animal forages.

Effects of insecticides on mortality, growth and bioaccumulation in black soldier fly (Hermetia illucens) larvae.

Residues of persistent insecticides may be present in the substrates on which insects are reared for food and feed, which may affect insect growth or survival. In addition, insecticidal substances may bio-accumulate in reared insects. The objective of this study was to assess potential effects of selected insecticides on the growth and survival of black soldier fly larvae (BSFL, Hermetia illucens) and on their safety when used as animal feed. Six insecticides (chlorpyrifos, propoxur, cypermethrin, imidacloprid, spinosad, tebufenozide) with different modes of action were tested in two sequential experiments. Cypermethrin was also tested with the synergist piperonyl butoxide (PBO). Standard BSFL substrate was spiked to the respective maximum residue level (MRL) of each insecticide allowed by the European Union to occur in feed; and BSFL were reared on these substrates. Depending on the observed effects in the first experiment, spiked concentrations tested in the second experiment were increased or reduced. At the concentrations applied (1 and 10 times MRL), three of the six tested substances (chlorpyrifos, propoxur, tebufenozide) did not affect the survival or biomass growth of BSFL, compared to the control (non-spiked) treatments. At MRL, imidacloprid stimulated the growth of BSFL compared to the controls. Spinosad and cypermethrin at the MRL level negatively affected growth and survival. The effects of cypermethrin appeared to be augmented by addition of PBO. A mean bio-accumulation factor of ≤0.01 was found in both experiments for all substances-except for cypermethrin, which was comparatively high, but still below 1 (0.79 at 0.1 mg/kg). The lack of accumulation of insecticides in the larvae suggests that there is no risk of larval products being uncompliant with feed MRLs. However, we conclude that insecticides present in substrates may affect growth and survival of BSFL. More research on a larger variety of substances and insect species is recommended.

Toxic effects of dietary copper and EGCG on bioaccumulation, antioxidant enzyme and immune response of Korean bullhead, Pseudobagrus fulvidraco.

There are few reports of dietary Cu (copper) toxicity to Korean bullhead, Pseudobagrus fulvidraco, and little is known about recovery from dietary Cu exposure. In this study, P. fulvidraco (mean length 16.9 ± 1.38 cm, and mean weight 53.2 ± 1.22 g) were exposed for 4 weeks to dietary Cu concentration of 0 (control), 700, 900, and 1100 mg Cu kg-1 dry feed to establish maximum tolerable levels of dietary Cu. All fish were then fed the dietary EGCG (Epigallocatechin gallate) concentration of 100 and 500 mg EGCG kg-1 dry feed for a further 2 weeks to assess recovery. We were measured bioaccumulation (in the intestine, liver, and gill tissue), antioxidant enzymes (SOD and CAT) and immune responses (lysozyme and phagocytosis). The Cu exposure induced a significant accumulation in the intestine, liver, and gill tissues and the highest accumulation was observed in intestinal tissues (17-34 fold), but dietary EGCG exposure decreased (about 0.8-fold) Cu concentration in each tissue (ANOVA, P < 0.05). In antioxidant enzymes, SOD and CAT significantly increased by approximately 1.6-fold by dietary Cu exposure in the liver and gill tissue, respectively, but dietary EGCG exposure decreased SOD and CAT by about 1.1-fold, respectively (ANOVA, P < 0.05). For immune responses, lysozyme and phagocytosis in the blood significantly were decreased by approximately 1.5-fold, respectively, by dietary Cu exposure, but dietary EGCG exposure increased lysozyme and phagocytosis by about 1.1-fold, respectively (ANOVA, P < 0.05). During recovery period, bioaccumulation, antioxidant enzymes (SOD and CAT activity), and immune response (lysozyme and phagocytosis activity) tended to alleviate the significant changes by Cu exposure, and the tendency to return normal state was observed in high level of EGCG. The result of this study indicate that Cu exposure to P. fulvidraco affects bioaccumulation, antioxidant enzymes, and immune responses, and high level of EGCG were effective to alleviate the toxic effects of Cu exposure.

Rhizospheric pore-water content predicts the biochar-attenuated accumulation, translocation, and toxicity of cadmium to lettuce.

Metal bioavailability controls its behaviors in soil-plant system, especially involved in biochar amendment. This study compared a rhizospheric pore-water extraction against a BCR sequential extraction method to understand cadmium (Cd) bioavailability in two typical Chinese soils. Soils were spiked with five levels of Cd (CdCl2) and remediated with 3% corn-straw derived biochar. After 60 days of lettuce growth, Cd accumulation and enzyme activities in tissues were analyzed. Results showed that biochar increased soil properties (pH, CEC and SOM) compared to un-amended soils, but decreased contents of bioavailable Cd in soil pore-water (Cdpore-water) and BCR extracted Cd (CdFi+Fii). Contents of Cdpore-water were lower in yellow-brown soils than that in red soils. Pearson analysis showed that bioavailable Cd is negatively correlated with soil pH and CEC (p < 0.05). Cd accumulation in lettuce roots and leaves both were decreased by biochar addition, and the established linear equations proved that soil Cdpore-water is the best predictor for Cd accumulation in lettuce roots (r2 = 0.964) and in leaves (r2 = 0.953), followed by CdFi+Fii. Transfer factor (TF) values of Cd from roots to leaves were lower than 1, and slightly better correlated with soil Cdpore-water (r = -0.674, p < 0.01) than CdFi+Fii (r = -0.615, p < 0.01). Aggregated boosted tree (ABT) analyses indicated that soil properties together with Cdpore-water contribute more than 50% to root enzyme activities. Collectively, soil Cdpore-water is a promising predictor of Cd bioavailability, accumulation and toxicity in soil-plant system with biochar addition.

Phytotoxicity and accumulation of Cu in mature and young leaves of submerged macrophyte Hydrilla verticillata (L.f.) Royle.

Phytotoxicity and accumulation of Cu in mature and young leaves of submerged macrophyte Hydrilla verticillata (L.f.) Royle were investigated by analyzing the chlorophyll contents, chloroplast ultrastructure and leaf surface structure under different Cu treatments (0, 0.01, 0.05 and 0.1 f mg L-1). The results showed that 0.05 and 0.1 mg L-1 Cu treatment decreased the contents of Chl a and Chl b, and caused damage on leaf surface structure and chloroplast ultrastructure compared with control (0 mg L-1 Cu treatment). Higher concentration of Cu induced Chlorophyll decreases and the damages on the leaf surface structure and chloroplast ultrastructure were more pronounced in mature than in young leaves. It was observed that leaf Cu concentration increased almost linearly with exposure time and majority of the Cu accumulated in the cell walls. Among different cell wall fractions, the majority of Cu accumulated in cell walls was bound to the hemicellulose 1 and cellulose, followed by the pectin, hemicellulose 2. Mature leaves had significantly higher the concentrations of total Cu and bound-Cu in cell walls due to higher uronic acid content in their cell wall fractions (pectin, hemicellulose 1 and cellulose) than young leaves. Distinct cell wall composition might partially contribute to the different Cu toxicity and accumulation between mature and young leaves of submerged macrophyte H. verticillata. Our results show that mature leaves are more efficient in the uptake and accumulation of Cu than young leaves, which might explain why mature leaves sustain more severe damage.

Cadmium, chromium, nickel and nitrate accumulation in wheat (Triticum aestivum L.) using wastewater irrigation and health risks assessment.

The wastewater utilization for irrigation purposes is common practice in peri-urban areas located in vicinity of developed cities. This water contains elements like chromium (Cr), nickel (Ni), cadmium (Cd) and nitrate (NO3-N) that poses health risk when exposed to human. In this study effect of wastewater irrigation from Chakara wastewater plant, Faisalabad on growth of wheat and health risks was assessed. Pot experiment was conducted at Institute of Soil and Environmental, University of Agriculture, Faisalabad using different concentration of wastewater as treatment 100% tap water, 25% wastewater + 75% tap water, 50% wastewater + 50% tap water, 75% wastewater + 25% tap water, 100% wastewater. The results indicated that the wastewater irrigation negatively effects the plant growth and physiological parameters. The minimum plant height, grain weight, spike length, osmotic potential and SPAD values were recorded 50.33 cm, 1.47 g plant-1, 7.00 cm, 423 and 38.91 respectively in 100% wastewater irrigation. The risk quotient (RQ TEs) for each toxic element and cumulative risk index (RI TEs) values were calculated. The cadmium risk quotient (Cd RQ) for adults was on margin and value was >1 for in 75% wastewater + 25% tap water and 100% wastewater irrigation, while the RQ for Ni and Cr was <1. Maximum RI TEs values calculated in 100% wastewater irrigation 0.424 and 0.294 for children and adults respectively. Hence it was concluded that wastewater irrigation significantly increased the accumulation rate of metals and nitrate in wheat and cause potential health risks for children and adults.

Joint effects of naphthalene and microcystin-LR on physiological responses and toxin bioaccumulation of Landoltia punctata.

The co-contamination of naphthalene (NAP) and microcystin-LR (MC-LR) commonly occurs in eutrophic waters. However, the joint effects of NAP and MC-LR on plants in aquatic environments remain unknown. Landoltia punctata is characterized by high starch yields and high biomass in polluted waters and has been proven to be a bioenergy crop and phytoremediation plant. In this study, L. punctata was cultured in a nutrient medium with environmentally relevant NAP (0.1, 1, 3, 5, and 10 µg/L) and MC-LR (5, 10, 25, 50, and 100 µg/L) to determine individual and joint toxic effects. The effects of NAP and MC-LR on physiological responses of L. punctata, including growth, starch accumulation, and antioxidant responses, were studied. Bioaccumulation of MC-LR in L. punctata, with or without NAP, was also examined. The results showed that growth and chlorophyll-a contents of L. punctata were reduced at high concentrations of MC-LR (≥ 25 µg/L), NAP (≥ 10 µg/L) and their mixture (≥ 10 + 1 µg/L) after exposure for 7 d. Starch accumulation in L. punctata did not decrease when exposed to NAP and MC-LR, and higher starch content of 29.8 % ± 2.7 % DW could be due to the destruction of starch-degrading enzymes. The antioxidant responses of L. punctata were stronger after exposure to MC-LR + NAP than when exposed to a single pollutant, although not enough to avoid oxidative damage. NAP enhanced the bioaccumulation of MC-LR in L. punctata when NAP concentration was higher than 5 µg/L, suggesting that higher potentials of MC-LR phytoremediation with L. punctata may be observed in NAP and MC-LR co-concomitant waters. This study provides theoretical support for the application of duckweed in eutrophic waters containing organic chemical pollutants.

Proteomic analysis and biochemical alterations in marine mussel gills after exposure to the organophosphate flame retardant TDCPP.

Organophosphate flame retardants (OPFRs) are (re-)emergent environmental pollutants increasingly being used because of the restriction of other flame retardants. The chlorinated OPFR, tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is among those of highest environmental concern, but its potential effects in the marine environment have rarely been investigated. We exposed a widely used sentinel marine mussel species, Mytilus galloprovincialis, to 10 µg L-1 of TDCPP during 28 days and studied: (i) the kinetics of bioaccumulation and elimination of the compound, (ii) the effect on two molecular biomarkers, glutathione S-transferase (GST) and acetylcholinesterase (AChE) activities, and (iii) proteomic alterations in the gills, following an isobaric labeling quantitative shotgun proteomic approach, at two exposure times (7 and 28 days). Uptake and elimination of TDCPP by mussels were very fast, and the bioconcentration factor of this compound in mussels was 147 L kgww-1, confirming that this compound is not very bioaccumulative, as predicted by its chemical properties. GST activity was not affected by TDCPP exposure, but AChE activity was inhibited by TDCPP at both 7 and 28 days of exposure. Proteomic analysis revealed subtle effects of TDCPP in mussel gills, since few proteins (less than 2 % of the analysed proteome) were significantly affected by TDCPP, and effect sizes were low. The most relevant effects detected were the up-regulation of epimerase family protein SDR39U1, an enzyme that could be involved in detoxification processes, at both exposure times, and the down-regulation of receptor-type tyrosine-protein phosphatase N2-like (PTPRN2) after 7 days of exposure, which is involved in neurotransmitter secretion and might be related to the neurotoxicity described for this compound. Exposure time rather than TDCPP exposure was the most important driver of protein abundance changes, with 33 % of the proteome being affected by this factor, suggesting that stress caused by laboratory conditions could be an important confounding factor that needs to be controlled in similar ecotoxicology studies. Proteomic data are available via ProteomeXchange with identifier PXD019720.

Accumulation of paralytic shellfish toxins by Southern Rock lobster Jasus edwardsii causes minimal impact on lobster health.

Recurrent dinoflagellate blooms of Alexandrium catenella expose the economically and ecologically important Southern Rock Lobster in Tasmania to paralytic shellfish toxins (PST), and it is unknown if PST accumulation adversely affects lobster performance, health and catchability. In a controlled aquaculture setting, lobsters were fed highly contaminated mussels to accumulate toxin levels in the hepatopancreas (mean of 6.65 mg STX.2HCl equiv. kg-1), comparable to those observed in nature. Physiological impact of PST accumulation was comprehensively assessed by a range of behavioural (vitality score, righting ability and reflex impairment score), health (haemocyte count, bacteriology, gill necrosis and parasite load), nutritional (hepatopancreas index and haemolymph refractive index) and haemolymph biochemical (21 parameters including electrolytes, metabolites, and enzymes) parameters during a 63 day period of uptake and depuration of toxins. Exposure to PST did not result in mortality nor significant changes in the behavioural, health, or nutritional measures suggesting limited gross impact on lobster performance. Furthermore, most haemolymph biochemical parameters measured exhibited no significant difference between control and exposed animals. However, the concentration of potassium in the haemolymph increased with PST, whilst the concentration of lactate and the sodium:potassium ratio decreased with PST. In addition, exposed lobsters showed a hyperglycaemic response to PST exposure, indicative of stress. These findings suggest that PST accumulation results in some measurable indicators of stress for lobsters. However, these changes are likely within the adaptive range for Jasus edwardsii and do not result in a significant impairment of gross performance. Our findings support previous conclusions that crustaceans are relatively tolerant to PST and the implications for the lobster fishery are discussed.

Influence of environmentally relevant concentrations of Zn, Cd and Ni and their binary mixtures on metal uptake, bioaccumulation and development in larvae of the purple sea urchin Strongylocentrotus purpuratus.

Metal accumulation, disturbance of Ca2+ homeostasis, and occurrence of abnormalities are well-established consequences of single metal exposure during early development stages of sea urchins. However, the effects caused by low concentrations of metals and metal mixtures need to be better understood in marine invertebrates. Therefore, the present study investigated the effects of environmentally relevant concentrations of Zn (9 µg/L), Cd (30 µg/L) and Ni (5 µg/L) in single and binary exposures (Zn + Cd, Cd + Ni and Ni + Zn) to the early life stages of the purple sea urchin Strongylocentrotus purpuratus. Endpoints checked in all treatments after 48-h exposure were unidirectional metal influx rates, bioaccumulation, and Ca2+ influx rates. Additionally, the presence of abnormal larvae and developmental delay was evaluated at 24 h, 48 h and 72 h of exposure. Unidirectional influx rates of all three metals were significantly higher than control background rates in all single exposures and binary mixtures, and were generally not different between them. Net accumulation (body burden) of both Zn and Cd increased significantly as a result of their respective single exposures, while Ni accumulation decreased considerably. When Zn or Cd were presented in binary exposures with other metals, the net accumulations of Zn or Cd were reduced relative to single exposures to these metals, whereas this did not occur for Ni accumulation. Thus, bioaccumulation proved to be a better metric than influx rate measurements to analyze metal competition at a whole organism level at these low metal concentrations. Short-term Ca2+ influx also did not appear to be a sensitive metric, as the measured rates did not vary among all single and binary exposures, with the exception of a lower rate in Ni + Zn binary exposure. A critical aspect observed was the relationship between bioaccumulation versus influx measurements, which proved positive for Cd, but negative for Zn and Ni, demonstrating possible capacities for both Zn and Ni regulation by sea urchin larvae. Increases in larval abnormalities relative to controls occurred only after binary mixtures, starting at 48 h exposure and maintained until 72 h. However, delay of the sea urchin development by the presence of gastrula stage at 72 h exposure occurred in Zn and Ni single exposures and all metal mixtures, with very high abnormal development when Ni was present.

Alleviation of cadmium-induced phytotoxicity and growth improvement by exogenous melatonin pretreatment in mallow (Malva parviflora) plants.

The present work was aimed to study the effect of melatonin pretreatments on growth, oxidative stress modulation, cadmium (Cd) accumulation, and tolerance in mallow (Malva parviflora, Malvaceae) plants under the hydroponic system. Application of substances that can modulate the harmful effects of Cd on plant yield and reduce its accumulation in the edible parts is of particular importance. Therefore, the mallow plants pretreated with 15, 50, and 100 µM of melatonin were exposed to 50 µM Cd. Our results showed that melatonin, especially at 15 and 50 µM, led to positive effects on Cd tolerance, including a significant increase in growth, photosynthetic pigments, and soluble protein content. Exogenous melatonin could improve relative water content (RWC) and stomatal conductance in the plants treated with Cd, probably through an increase in proline. Further, lower concentrations of melatonin led to a decrease in Cd translocation to the shoots. Based on the results, melatonin considerably increased catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (GPX) activities as well as the production of phenols. The increased activity of antioxidant enzymes led to a decrease in electrolyte leakage (EL), lipid peroxidation, and H2O2 content in the plants exposed to Cd stress. Under Cd stress, the increased phenols content in melatonin-pretreated plants could be due to the induction of phenylalanine ammonia-lyase (PAL) activity and an increase in shoot soluble carbohydrates. The results showed that the use of melatonin could reduce oxidative stress and improve biomass in the plants exposed to Cd. At least in our experimental conditions, this information appears to be useful for healthy food production.

Amelioration of hexavalent chromium-induced bioaccumulation, oxidative stress, tight junction proteins and immune-related signaling factors by Allium mongolicum Regel flavonoids in Ctenopharyngodon idella.

Chromium (Cr) is the most common heavy metal and that becomes toxic when present at higher concentrations in aquatic environments. Allium mongolicum Regel flavonoids (AMRF) has been documented to possess detoxification, antioxidant and anti-inflammatory properties. The aim of this study was to explore the potential of dietary AMRF and Cr exposure on bioaccumulation, oxidative stress, and immune response in Ctenopharyngodon idella. After acclimation, 360 fish were randomly distributed into six groups. The fish were fed with diets supplemented with Cr and/or AMRF for 4 weeks (28 days), the Cr concentrations were 0, 120, and 240 mg/kg and the concentrations of AMRF were 0 or 40 mg/kg, respectively. The results shown that Cr accumulation in the kidney, liver, spleen, intestine and gill were significantly increased following Cr exposure, dietary AMRF supplementation attenuated the increased in Cr accumulation. Dietary AMRF supplementation significantly reduced the levels of malondialdehyde (MDA) and protein carbonyl (PC) in liver, spleen and gill compared with the same Cr dose groups. When fish were supplemented with AMRF significantly increased lysozyme activity (LZM), complement 3 (C3) in kidney and intestine compared with the same Cr dose groups. Serum glutamic oxalate transaminase (GOT) and glutamic pyruvate transaminase (GPT) were significantly increased following exposure to Cr. Dietary AMRF supplementation significantly decreased GOT and GPT activity in the serum. In addition, AMRF supplementation can decrease the expression of inflammatory (NF-κB p65, TNF-α and IL-1ß) and increased the expression of tight junction proteins (occludin and ZO-1) following Cr exposure in C. idella. These results indicate that AMRF has the potential to alleviate the effects of Cr toxicity in C. idella.

Effect on intestinal microbiota, bioaccumulation, and oxidative stress of Carassius auratus gibelio under waterborne cadmium exposure.

Cadmium (Cd) is a hazardous pollutant known to exert various toxic effects and other sublethal effects on aquatic organisms, and induce a variety of adverse effects on human health, and can be commonly found in environment. The aim of this study was to explore the effects of waterborne Cd exposure on the intestinal microbiota, and Cd accumulation and oxidative response in Carassius auratus gibelio (C. gibelio). The fish were exposed to waterborne Cd at 0, 1, 2, and 4 mg/L for 30 days. Waterborne Cd exposure resulted marked alterations in the composition of microbiota. At the genus level, Bacteroides, Aeromonas, Akkermansia, Acinetobacter, Chryseobacterium, Shinella, Cetobacterium and Bacillus were significantly changed in Cd groups. The results obtained indicate that Cd exposure significantly increased the bioaccumulation level of Cd and profoundly affected antioxidant enzyme including superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), total antioxidant (T-AOC), malonaldehyde (MDA), and glutathione-S-transferase (GST). The present findings may provide a new framework for the role of gut microbiota in the response to environmental chemical contamination and in the pathogenesis of body disorders.

In Vitro Biotransformation Assays Using Liver S9 Fractions and Hepatocytes from Rainbow Trout (Oncorhynchus mykiss): Overcoming Challenges with Difficult to Test Fragrance Chemicals.

In vitro metabolic stability assays using rainbow trout (Oncorhynchus mykiss) isolated hepatocytes (RT-HEP) or hepatic S9 fractions (RT-S9) were introduced to provide biotransformation rate data for the assessment of chemical bioaccumulation in fish. The present study explored the suitability of the RT-HEP and RT-S9 assays for difficult test chemicals, and the in vitro-based predictions were compared to in silico-based predictions and in vivo-measured bioconcentration factors (BCFs). The results show that volatile or reactive chemicals can be tested with minor modifications of the in vitro protocols. For hydrophobic chemicals, a passive dosing technique was developed. Finally, a design-of-experiment approach was used to identify optimal in vitro assay conditions. The modified assay protocols were applied to 10 fragrances with diverse physicochemical properties. The in vitro intrinsic clearance rates were higher in the S9 than in the hepatocyte assay, but the in vitro-in vivo (IVIV) predictions were comparable between the 2 assays. The IVIV predictions classified the test chemicals as nonbioaccumulative (BCF < 2000), which was in agreement with the in vivo data but in contrast to the in silico-based predictions. The findings from the present study provide strong evidence that the RT-HEP and RT-S9 assays can provide reliable estimates of in vivo biotransformation rates for test chemicals with difficult physicochemical properties. Environ Toxicol Chem 2020;39:2396-2408. © 2020 SETAC.

Bioconcentration of 2,4,6-tribromophenol (TBP) and thyroid endocrine disruption in zebrafish larvae.

2,4,6-tribromophenol (TBP) is generally used as a brominated flame retardant but is produced in the degradation of tetrabromobisphenol-A. Although TBP is frequently detected in the environment and in various biota, including fish species, we still know little about its toxicity and environmental health risk. Here we investigated the bioconcentration and effects of TBP on the thyroid endocrine system by using zebrafish as a model. Zebrafish embryos (2 h post-fertilization, hpf) were exposed to five concentrations of TBP (0, 0.3, 1, 10, and 100 µg/L) until 144 hpf. According to our chemical analysis, TBP underwent bioconcentration in zebrafish larvae. However, acute exposure to TBP did not affect the hatching of embryos or their risk of malformation, nor the growth and survival of larvae, indicating low developmental toxicity of TBP. The whole-body thyroxine (T4) contents were significantly increased in zebrafish larvae after exposure to TBP, indicating thyroid endocrine disruption occurred. Gene transcription levels in the hypothalamic-pituitary-thyroid (HPT) axis were also examined in larvae; these results revealed that the transcription of corticotrophin-releasing hormone (crh), thyrotropin-releasing hormone (trh), and thyroid-stimulating hormone (tshß) were all significantly downregulated by exposure to TBP. Likewise, genes encoding thyronine deiodinases (dio1, dio2, and dio3a/b) and thyroid hormone receptors (trα and trß) also had their transcription downregulated in zebrafish. Further, the gene transcription and protein expression of binding and transport protein transthyretin (TTR) were significantly increased after TBP exposure. Taken together, our results suggest the bioavailability of and potential thyroid endocrine disruption by TBP in fish.

Salicylic acid mediated reduction in grain cadmium accumulation and amelioration of toxicity in Oryza sativa L. cv Bandana.

Contamination of agricultural fields with Cadmium (Cd) due to several agricultural practices is increasing worldwide. The rice plants can easily take up Cd and accumulate it into different parts, including the grains, posing a threat to human health even at low concentration exposure. Several phytohormones, including Salicylic acid (SA) have been investigated since long for its alleviating properties under various biotic and abiotic stress conditions. In the present study, 100 µM SA application to ameliorate 25 µM Cd stress was studied for 72 h in hydroponics in Oryza sativa cv. Bandana seedlings. Pot experiments were done with same treatment condition and plants were grown till maturity. SA application to Cd exposed rice seedlings alleviated the stress condition, which was established by several physiological, biochemical, histochemical and gene expression analysis. SA treatment to Cd stressed seedlings showed elevated photosynthetic pigment content, on-protein thiol content and relieved the Cd induced growth inhibition considerably. It lowered the accumulation of ROS like, O2- and H2O2 with a regulated antioxidative enzymatic activity. SA application in Cd exposed rice seedlings had upregulated expression of OsHMA3 and OsPCS1 whereasOsNRAMP2 gene was downregulated. Co-application of SA and Cd led to higher yield and improved agronomic traits in comparison to only Cd exposed plants under pot experimentation. Daily intake of Cd and Carcinogenic risk were also reduced by 99.75% and 99.99% respectively in the SA treated Cd stressed plants. SA positively affected the growth and tolerance of rice seedlings to Cd stress. Hence, SA addition to Cd contaminated soil can ensure rice cultivation without posing health risk to consumers.

Analysis of the effect of cadmium stress on root exudates of Sedum plumbizincicola based on metabolomics.

Root exudates are the most direct manifestation of the response of plants changes in the external environment. Therefore, based on non-targeted gas chromatography-time-of-flight mass spectrometry and metabolomics, the response of Sedum plumbizincicola root exudates to Cd stress was used to reveal the possible mechanism of resistance to or accumulation of Cd. The results showed that Cd significantly changed the composition and contents of S. plumbizincicola root exudates. A total of 155 metabolites were identified in S. plumbizincicola root exudates, among which 33 showed significant differences under Cd stress, including organic acids, amino acids, lipids, and polyols. Cd stress suppressed organic acid metabolism and lipid metabolism in S. plumbizincicola and significantly affected amino acid metabolism. There were 16 metabolic pathways related to Cd stress, among which arginine and proline metabolism, valine, leucine, and isoleucine biosynthesis, glycine, serine, and threonine metabolism, glutathione metabolism, and purine metabolism were the key pathways with the highest correlation, and were closely related to the stress resistance of S. plumbizincicola.

Do environmentally relevant concentrations of fluoxetine and citalopram impair stress-related behavior in zebrafish (Danio rerio) embryos?

Selective serotonin reuptake inhibitors (SSRIs) have been shown to interfere with various physiological functions of aquatic organisms, yet the neuroactive potential of low concentrations of SSRIs in the aquatic environment is unclear. The current study investigated the effects of fluoxetine and citalopram on the visual motor response (VMR) of 107 h old zebrafish (Danio rerio) embryos. Results document a reduction in stress-related swimming activity of zebrafish embryos at environmentally relevant concentration levels, with fluoxetine being more effective than citalopram. Further experiments were designed to elucidate (1) if the lower neuroactive potential of citalopram is due to differences in uptake kinetics, (2) if the metabolite of fluoxetine, norfluoxetine, contributes to the neuroactive potential of fluoxetine, (3) and how SSRIs and their metabolites interact in equimolar mixtures. At the stage of 120 h, zebrafish embryos accumulate citalopram at significantly lower rates (up to 127 times) than fluoxetine. Moreover, it was demonstrated that norfluoxetine reduces the embryonic VMR similarly to fluoxetine resulting in additive effects of these substances on stress-related behavior in zebrafish embryos. In contrast, the interaction of fluoxetine, norfluoxetine and citalopram varied with test concentrations of the equimolar mixtures. Findings provide evidence that environmentally relevant concentrations of fluoxetine reduce stress-related behavior of zebrafish embryos, while these effects may be enhanced by the interaction of multiple SSRIs and their metabolites in environmental exposure scenarios.

Deciphering lead tolerance mechanisms in a population of the plant species Biscutella auriculata L. from a mining area: Accumulation strategies and antioxidant defenses.

The uptake and distribution of Pb and the mechanisms involved in the metal tolerance have been investigated in a mine population of Biscutella auriculata. Seedlings were exposed to 125 µM Pb(NO3)2 for 15 days under semihydroponic conditions. The results showed an increase in the size of Pb-treated seedlings and symptoms of toxicity were not observed. ICP-OES analyses showed that Pb accumulation was restricted to root tissue. Imaging of Pb accumulation by dithizone histochemistry revealed the presence of the metal in vacuoles and cell wall in root cells. The accumulation of Pb in vacuoles could be stimulated by an increase in phytochelatin PC2 content. Pb did not promote oxidative damage and this is probably due the increase of antioxidative defenses. In the leaves, Pb produced a significant increase in superoxide dismutase activity, while in roots an increase in catalase and components of the Foyer- Halliwell-Asada cycle were observed. The results indicated that Biscutella auriculata has a high capacity to tolerate Pb and this is mainly due to a very efficient mechanism to sequester the metal in roots and a capacity to avoid oxidative stress. This species could therefore be very useful for phytostabilization and repopulation of areas contaminated with Pb.

Effects of lead, cadmium and zinc on protein changes in Silene vulgaris shoots cultured in vitro.

In the present research, Silene vulgaris as a representative species growing on both unpolluted and heavy metal (HM) polluted terrains were used to identify ecotype-specific responses to metallic stress. Growth, cell ultrastructure and element accumulations were compared between non-metallicolous (NM), calamine (CAL) and serpentine (SER) specimens untreated with HMs and treated with Pb, Cd and Zn ions under in vitro conditions. Moreover, proteins' modifications related to their level, carbonylation and degradations via vacuolar proteases were verified and linked with potential mechanisms to cope with ions toxicity. Our experiment revealed diversified strategy of HM uptake in NM and both metallicolous ecotypes, in which antagonistic relationship of Zn and Pb/Cd ions provided survival benefits for the whole organism. Despite this similarity, growth rate and metabolic pathways induced in CAL and SER shoots varied significantly. Exposition to HMs in CAL culture led to drop in protein level by approximately 16% compared to the control. This parameter nearly correlated with the enhanced activity of proteases at pH 5.2 as well as possible glutamate changes to proline and reduced glutathione, resulting in intensified growth and first signs of cell senescence. In turn, SER shoots were characterized by growth retardation (to 53% of the control), although protein level and carbonylation were not modified, while a deeper insight into protein network showed its remodeling towards production of polyamines and 2-oxoglutarate delivered to the Krebs cycle. Contrary, an uncontrolled HM influx in NM shoots contributed to morpho-structural disorders accompanied by an increase activity of proteases involved in the degradation of oxidized proteins, what pointed to metal-induced autophagy. Taken together, S. vulgaris ecotypes respond to stress by triggering various mechanisms engaged their survival and/or death under HM treatment.