Waterborne Zn Exposure Research Articles

Meeting Zn Needs during Medaka Eye Development: Nanoscale Visualization of Retina by Expansion Microscopy.

Fish eyes require high Zn levels to support their early development. Although numerous studies have been conducted on the nutritional and toxic effects of Zn on the eye, the Zn requirement for retinal cell development is still debatable. Moreover, due to the complexity of the retinal structure, it is difficult to clearly visualize each retinal layer and accurately separate cell morphology in vivo by conventional methods. In the present study, we for the first time have achieved nanoscale imaging of retinal anatomy affected by dietary and waterborne Zn exposure by novel expansion microscopy. We demonstrated that the fish retina showed different developmental strategies in response to dietary and aqueous Zn exposures. Excess dietary Zn produced toxicity to retinal photoreceptor cells, resulting in a reduction in cell number and cell area, and this toxicity became severe with biological development. In contrast, waterborne Zn in the natural environment probably failed to meet the Zn requirements of retinal development. Overall, our results indicated that during early development, the Zn requirement of the fish eyes was sensitive, and oversupplementation led to impaired photoreceptor cell development. Our study has provided new perspectives using the powerful and novel expansion microscopy technique in toxicity assessment, enabling ultra-clear visualization of small but complex organ development.

Waterborne zinc bioaccumulation influences glucose metabolism in orange-spotted grouper embryos

Fish embryos, as an endogenous system, strictly regulate an energy metabolism that is particularly sensitive to environmental pressure. This study used orange-spotted grouper embryos and stable isotope 67Zn to test the hypothesis that waterborne Zn exposure had a significant effect on energy metabolism in embryos. The fish embryos were exposed to a gradient level of waterborne 67Zn, and then sampled to quantify 67Zn bioaccumulation and mRNA expressions of key genes involved glucose metabolism. The results indicated that the bioaccumulated 67Zn generally increased with increasing waterborne 67Zn concentrations, while it tended to be saturated at waterborne 67Zn > 0.7 mg L−1. As we hypothesized, the expression of PK and PFK gene involved glycolysis pathway was significantly up-regulated under waterborne 67Zn exposure >4 mg L−1. Waterborne 67Zn exposure >2 mg L−1 significantly suppressed PCK and G6PC gene expression involved gluconeogenesis pathway, and also inhibited the AKT2, GSK-3beta and GLUT4 genes involved Akt signaling pathway. Our findings first characterized developmental stage-dependent Zn uptake and genotoxicity in fish embryos. We suggest fish embryos, as a small-scale modeling biosystem, have a large potential and wide applicability for determining cytotoxicity/genotoxicity of waterborne metal in aquatic ecosystem.

Transcriptome analysis reveals differential gene expression in Lateolabrax maculatus following waterborne Zn exposure

The spotted sea bass, Lateolabrax maculatus, is an economically important farmed fish species in the world. To gain a better understanding of the immune response, antioxidant enzymes and stress (heat shock protein 70, HSP70) in L. japonicas, we analyzed its transcriptome following waterborne Zn exposure (35 mg/L Zn) at 24 h in the liver. Following assembly and annotation, 104,787 unigenes with an average length of 2230 bp were identified. A total of 2882 differentially expressed genes (DEGs) in the L. japonicas were observed at 24 h post waterborne zinc treatment, including 1289 up-regulated genes and 1593 down-regulated genes. Clusters of Orthologous Groups of proteins (KOG/COG) annotation demonstrated that a total of 32,866 unigenes classified into 26 categories. Gene ontology (GO) analysis revealed 20 biological process subcategories, 4 cellular component sub-categories and 17 molecular function subcategories. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis identified immune, antioxidant and stress pathways. Quantitative reverse transcription polymerase chain reaction measured the expression of 10 genes involved in the immune, antioxidant and stress. HSP70 was significantly up-regulated, whereas g-type lysozyme (LYZ-G), CU/ZN superoxide dismutase (CU/ZN-SOD), glutathione peroxidase 7 (GPX7), glutathione reductase (GR), glutathione-S-transferase (GST), nuclear factor erythroid 2-related factor 2 (Nrf2), Kelch-like ECH-associated protein-1 (Keap1) were significantly down-regulated. This study enriches the L. japonicas transcriptome database and provides insight into the immune response, antioxidants and stress of L. japonicas against heavy metal pollution.

Effects of acute and chronic zinc exposure on steroid hormone receptor expression in the lined seahorse, Hippocampus erectus

The element zinc (Zn) can elicit a variety of physiological responses in animals, particularly through its effects on steroid hormone receptor expression. Here, we evaluated the effects of Zn on the expression profiles of steroid hormone receptors in the lined seahorse, Hippocampus erectus, following experimental acute and chronic Zn exposure. Complex response patterns were observed; however, steroid hormone receptor expression in liver and kidney following 12‐day Zn injection (acute exposure) differed significantly from the control treatments. Apparent effects on hepatic lipid metabolism and osmotic regulation in the kidney need detailed investigation. Furthermore, 49‐day (chronic) waterborne Zn exposure produced a significant decrease in both estrogen receptor (ER) expression levels and estradiol concentrations. In additional, chronic exposure led to a substantial reduction of hepatic adipose cells, implying that Zn may regulate hepatic lipid metabolism via ER and estradiol. Our study demonstrated the effects of both acute and chronic Zn exposure on expression patterns of steroid hormone receptors, which will contribute to understanding the mechanism of Zn‐induced hepatic lipid metabolism and other physiological responses in seahorses.

Zn Stimulates the Phospholipids Biosynthesis via the Pathways of Oxidative and Endoplasmic Reticulum Stress in the Intestine of Freshwater Teleost Yellow Catfish.

The hypothesis of our study was that waterborne Zn exposure evoked phospholipids (PL) biosynthesis to compensate for the loss of membrane integrity, and the pathways of oxidative stress and endoplasmic reticulum (ER) stress mediated the Zn-evoked changes of PL biosynthesis. Thus, we conducted RNA sequencing to analyze the differences in the intestinal transcriptomes between the control and Zn-treated P. fulvidraco. The 56-day Zn exposure increased the intestinal Zn accumulation, and mRNA levels of 816 genes were markedly up-regulated, while that of 263 genes were down-regulated. Many differentially expressed genes in the pathways of PL biosynthesis and protein processing in ER were identified. Their expression profiles indicated that waterborne Zn exposure injured protein metabolism, induced PL biosynthesis caused oxidative stress and ER stress, and activated the unfolded protein response. Then, using the primary enterocytes, we identified the mechanism of oxidative and ER stress mediating Zn-induced PL biosynthesis, and indicated that the activation of these pathways constituted adaptive mechanisms to reduce Zn toxicity. Our study demonstrated that Zn exposure via the water increased Zn accumulation and PL biosynthesis, and that oxidative stress and ER stress were interdependent and mediated the Zn-induced PL biosynthesis of the intestine in the freshwater teleost.

Characterization of twelve autophagy-related genes from yellow catfish Pelteobagrus fulvidraco and their transcriptional responses to waterborne zinc exposure

Autophagy acts as important cytoprotective mechanism in response to adverse environment conditions. The hypothesis of the present study is that autophagy acts as protective responses to waterborne Zn exposure. To this end, the full-length cDNA sequences of 12 key genes related to autophagy in yellow catfish Pelteobagrus fulvidraco were cloned, and their mRNA expression profiles and transcriptional responses to waterborne Zn exposure were explored. The 12 genes (SQSTM1, Beclin1, ULK1A, ULK1B, ATG13-1, ATG13-2, ATG101, ATG9A, ATG9B, ATG3, ATG5 and ATG7) mediated the core autophagy machinery, including autophagosome membrane initiation, nucleation, expansion, closure and maturation. All of these members shared similar domain structure to their orthologous genes of other vertebrates. Their mRNAs were widely expressed in various tissues, but at different levels. Zn exposure increased the amount of hepatic autophagic vacuoles, and elevated the mRNA levels of SQSTM1, ULK1A, ULK1B, ATG13, ATG101, ATG9A, ATG9B, ATG3 and ATG7 in a dose- and time-dependent manner, indicating autophagy activation. These results indicated that autophagy acted as an adaptive response to protect from Zn toxicity and confirmed our hypothesis. Moreover, those up-regulated genes may play crucial roles in autophagy response to Zn exposure. For the first time, we characterized the full-length cDNA sequences of twelve autophagy related genes from fish, and determined their transcriptional responses to waterborne Zn exposure, which would contribute to our understanding of the molecular basis of autophagy and Zn toxicity, and also shed new insights on the potential role of autophagy as an adaptive response against metal toxicity in vertebrates.

Identification of apoptosis-related genes Bcl2 and Bax from yellow catfish Pelteobagrus fulvidraco and their transcriptional responses to waterborne and dietborne zinc exposure

Apoptosis plays a key role in the physiology of multicellular organisms, and has been well studied in mammals, but not in teleosts. Zinc (Zn) has been shown to be an important regulator of apoptosis and apoptosis involves in the regulation of lipid metabolism. Moreover, our recent study indicated that waterborne and dietborne Zn exposure differently influenced lipid metabolism in Pelteobagrus fulvidraco, but further mechanism remained unknown. The hypothesis of the present study is that apoptosis mediated the Zn-induced changes of lipid metabolism of P. fulvidraco subjected to different exposure pathways. To this end, we cloned full-length cDNA sequences of Bcl2 and three Bax subtypes involved in apoptosis in P. fulvidraco, explored their mRNA expressions in responses to different Zn exposure pathways. Bcl2 and three Bax subtypes shared similar domain structure as typical pro- and anti-apoptotic Bcl2 family members. Their mRNAs were widely expressed among various tissues, but at variable levels. Waterborne Zn exposure down-regulated mRNA levels of Baxg and ratios of Baxa/Bcl2, and Baxg/Bcl2, but showed no significant effects on mRNA abundances of Bcl2, Baxa and Baxb, and the ratio of Baxb/Bcl2. In contrast, dietborne Zn exposure up-regulated mRNA levels of Bcl2, Baxa, Baxb and Baxg, but reduced the ratios of Baxa/Bcl2, Baxb/Bcl2, and Baxg/Bcl2. Considering their important roles of these genes in apoptosis induced by Zn, apoptosis may mediate the Zn-induced changes of hepatic lipid metabolism of Pelteobagrus fulvidraco under different Zn exposure pathways. For the first time, we characterized the full-length cDNA sequences of Bcl2 and three Bax subtypes, determined their expression profiles and transcriptional responses to different Zn exposure pathways, which would contribute to our understanding of the molecular basis of apoptosis, and also provide new insights into physiological responses to different Zn exposure pathways.

Endoplasmic reticulum (ER) stress and cAMP/PKA pathway mediated Zn-induced hepatic lipolysis

The present study was performed to determine the effect of Zn exposure influencing endoplasmic reticulum (ER) stress, explore the underlying molecular mechanism of Zn-induced hepatic lipolysis in a fish species of significance for aquaculture, yellow catfish Pelteobagrus fulvidraco. We found that waterborne Zn exposure evoked ER stress and unfolded protein response (UPR), and activated cAMP/PKA pathway, and up-regulated hepatic lipolysis. The increase in ER stress and lipolysis were associated with activation of cAMP/PKA signaling pathway. Zn also induced an increase in intracellular Ca2+ level, which could be partially prevented by dantrolene (RyR receptor inhibitor) and 2-APB (IP3 receptor inhibitor), demonstrating that the disturbed Ca2+ homeostasis in ER contributed to ER stress and dysregulation of lipolysis. Inhibition of ER stress by PBA attenuated UPR, inhibited the activation of cAMP/PKA pathway and resulted in down-regulation of lipolysis. Inhibition of protein kinase RNA-activated-like ER kinase (PERK) by GSK2656157 and inositol-requiring enzyme (IRE) by STF-083010 differentially influenced Zn-induced changes of lipid metabolism, indicating that PERK and IRE pathways played different regulatory roles in Zn-induced lipolysis. Inhibition of PKA by H89 blocked the Zn-induced activation of cAMP/PKA pathway with a concomitant inhibition of ER stress-mediated lipolysis. Taken together, our findings highlight the importance of the ER stress–cAMP/PKA axis in Zn-induced lipolysis, which provides new insights into Zn toxicology in fish and probably in other vertebrates.

Role and mechanism of the AMPK pathway in waterborne Zn exposure influencing the hepatic energy metabolism of Synechogobius hasta.

Previous studies have investigated the physiological responses in the liver of Synechogobius hasta exposed to waterborne zinc (Zn). However, at present, very little is known about the underlying molecular mechanisms of these responses. In this study, RNA sequencing (RNA-seq) was performed to analyse the differences in the hepatic transcriptomes between control and Zn-exposed S. hasta. A total of 36,339 unigenes and 1,615 bp of unigene N50 were detected. These genes were further annotated to the Nonredundant protein (NR), Nonredundant nucleotide (Nt), Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups (COG) and Gene Ontology (GO) databases. After 60 days of Zn exposure, 708 and 237 genes were significantly up- and down-regulated, respectively. Many differentially expressed genes (DEGs) involved in energy metabolic pathways were identified, and their expression profiles suggested increased catabolic processes and reduced biosynthetic processes. These changes indicated that waterborne Zn exposure increased the energy production and requirement, which was related to the activation of the AMPK signalling pathway. Furthermore, using the primary hepatocytes of S. hasta, we identified the role of the AMPK signalling pathway in Zn-influenced energy metabolism.

Effects of salinity on short-term waterborne zinc uptake, accumulation and sub-lethal toxicity in the green shore crab (Carcinus maenas)

Waterborne zinc (Zn) is known to cause toxicity to freshwater animals primarily by disrupting calcium (Ca) homeostasis during acute exposure, but its effects in marine and estuarine animals are not well characterized. The present study investigated the effects of salinity on short-term Zn accumulation and sub-lethal toxicity in the euryhaline green shore crab, Carcinus maenas. The kinetic and pharmacological properties of short-term branchial Zn uptake were also examined. Green crabs (n=10) were exposed to control (no added Zn) and 50μM (3.25mgL−1) of waterborne Zn (∼25% of 96h LC50 in 100 seawater) for 96h at 3 different salinity regimes (100%, 60% and 20% seawater). Exposure to waterborne Zn increased tissue-specific Zn accumulation across different salinities. However, the maximum accumulation occurred in 20% seawater and no difference was recorded between 60% and 100% seawater. Gills appeared to be the primary site of Zn accumulation, since the accumulation was significantly higher in the gills relative to the hepatopancreas, haemolymph and muscle. Waterborne Zn exposure induced a slight increase in haemolymph osmolality and chloride levels irrespective of salinity. In contrast, Zn exposure elicited marked increases in both haemolymph and gill Ca levels, and these changes were more pronounced in 20% seawater relative to that in 60% or 100% seawater. An in vitro gill perfusion technique was used to examine the characteristics of short-term (1–4h) branchial Zn uptake over an exposure concentration range of 3–12μM (200–800μgL−1). The rate of short-term branchial Zn uptake did not change significantly after 2h, and no difference was recorded in the rate of uptake between the anterior (respiratory) and posterior (ion transporting) gills. The in vitro branchial Zn uptake occurred in a concentration-dependent manner across different salinities. However, the rate of uptake was consistently higher in 20% seawater relative to 60% or 100% seawater – similar to the trend observed with tissue Zn accumulation during in vivo exposure. The short-term branchial Zn uptake was found to be inhibited by lanthanum (a blocker of voltage-independent Ca channels), suggesting that branchial Zn uptake occurs via the Ca transporting pathways, at least in part. Overall, our findings indicate that acute exposure to waterborne Zn leads to the disruption of Zn and Ca homeostasis in green crab, and these effects are exacerbated at the lower salinity.

Effect of waterborne zinc exposure on lipid deposition and metabolism in hepatopancreas and muscle of grass carp Ctenopharyngodon idella.

The aim of the present study was to explore the effect of waterborne zinc (control, 0.85, 2.20, 3.10mg/l, respectively) exposure on lipid deposition and metabolism in the hepatopancreas and muscle of grass carp Ctenopharyngodon idella. The lipid content, Zn accumulation, and the activities and expression levels of several enzymes involved in lipid metabolism were determined in hepatopancreas and muscle. Waterborne Zn exposure reduced growth performance and increased Zn accumulation in both tested tissues. In hepatopancreas, Zn exposure increased lipid content, the activities of lipogenic enzymes, such as 6PGD, G6PD, ME, ICDH and FAS, as well as the mRNA expression level of G6PD, 6PGD, ICDH, FAS and SREBP-1. But the activity of CPT I and the mRNA expression of HSL, CPT Iα1a, CPT Iα2a and PPARα were down-regulated by Zn exposure. In contrast, in muscle, waterborne Zn exposure decreased lipid deposition, activities of 6GPD, ICDH and ME, as well as the mRNA expression level of G6PD, ICDH, ME, FAS and SREBP-1. However, the activity of CPT I as well as the mRNA expression level of PPARα, HSL, CPT Iα2a, CPT Iα1b and CPT Iβ were up-regulated by Zn exposure. Our results indicate that waterborne Zn increases lipid content by up-regulating lipogenesis and down-regulating lipolysis in hepatopancreas. But, in muscle, waterborne Zn reduces lipid accumulation by up-regulating lipolysis and down-regulating lipogenesis. Differential patterns of lipid deposition, enzymatic activities and genes' expression indicate the tissue-specific regulatory mechanism in fish.

Effect and mechanism of waterborne prolonged Zn exposure influencing hepatic lipid metabolism in javelin goby Synechogobius hasta.

The present study was conducted to determine the effect and mechanism of waterborne Zn exposure influencing hepatic lipid deposition and metabolism in javelin goby Synechogobius hasta. S. hasta were exposed to four waterborne Zn concentrations (Zn 0.005 [control], 0.18, 0.36 and 0.55 mg l(-1) , respectively) for 60 days. Sampling occurred at days 20, 40 and 60, respectively. Zn exposure increased Zn content, declined hepatic lipid content and reduced viscerosomatic and hepatosomatic indices and lipogenic enzyme activities, including 6-phosphogluconate dehydrogenase (6PGD), glucose-6-phosphate dehydrogenase (G6PD), malic enzyme (ME) and fatty acid synthase (FAS). At days 20 and 60, Zn exposure decreased hepatic mRNA levels of 6PGD, G6PD, ME, FAS, acetyl-CoA carboxylase (ACC)α, ACCβ, hormone-sensitive lipase (HSL)a, HSLb, sterol-regulator element-binding protein (SREBP)-1, peroxisome proliferators-activated receptor (PPAR)α and PPARγ. However, the mRNA levels of CPT 1 and adipose triglyceride lipase increased following Zn exposure. On day 40, Zn exposure reduced hepatic mRNA expression of 6PGD, G6PD, ME, FAS, ACCα, ACCβ, HSLa, HSLb, SREBP-1 and PPARγ but increased mRNA expression of CPT 1, adipose triglyceride lipase and PPARα. General speaking, Zn exposure reduced hepatic lipid content by inhibiting lipogenesis and stimulating lipolysis. For the first time, the present study provided evidence that chronic Zn exposure differentially influenced mRNA expression and activities of genes and enzymes involved in lipogenic and lipolytic metabolism in a duration-dependent manner, and provided new insight into the relationship between metal elements and lipid metabolism. Copyright © 2015 John Wiley & Sons, Ltd.

Dietary fenofibrate reduces hepatic lipid deposition by regulating lipid metabolism in yellow catfish Pelteobagrus fulvidraco exposed to waterborne Zn.

Fenofibrate is known to possess lipid-lowering effects by regulation of gene transcription involved in lipid metabolism. Waterborne Zn exposure induces lipid deposition in yellow catfish Pelteobagrus fulvidraco. Thus, the present working hypothesis is that dietary fenofibrate addition will reduce hepatic lipids in yellow catfish exposed to waterborne Zn. To this end, juvenile yellow catfish were exposed to 0.04 (control), 0.35 mg/L waterborne Zn, 0.15% dietary fenofibrate, and 0.35 mg Zn/l + 0.15% dietary fenofibrate for 8 weeks. Growth performance, lipid deposition and metabolism in the liver were determined. Dietary fenofibrate promoted growth performance and reduced hepatic lipid content of yellow catfish exposed to waterborne Zn. However, these effects did not appear in fish in normal water. The lipid-lowering effect of fenofibrate on fish exposed to waterborne Zn was associated with increased lipolysis, as indicated by increased CPT I activities and expression of lipolytic genes PPARα, CPT IA, ATGL and HSL, and with reduced lipogenesis as indicated by reduced activities of G6PD, 6PGD, ME and ICDH. Dietary fenofibrate significantly increased mRNA levels of FAS, LPL and ACCα, but reduced mRNA levels of ACCβ and PPARγ in fish exposed to waterborne Zn. Pearson correlations between transcriptional factors expression, and activities and expression of several enzymes were observed, indicating that changes at the molecular and enzymatic levels may underlie the patterns of lipid metabolism and accordingly affect hepatic fat storage. Taken together, our results suggest that the lipid-lowering effect of fenofibrate was attributed, in part, to the down-regulation of lipogenesis and up-regulation of fatty acid oxidation.

Different effect of dietborne and waterborne Zn exposure on lipid deposition and metabolism in juvenile yellow catfish Pelteobagrus fulvidraco

Juvenile yellow catfish Pelteobagrus fulvidraco were exposed to 0.04 or 0.35mgl−1 waterborne Zn, 27.25 or 213.84mgkg−1 dietary Zn, singly or in combination for 42 days. Growth and lipid metabolism in juvenile yellow catfish were investigated. Growth and survival were significantly inhibited by single waterborne Zn exposure but not by dietary Zn exposure. Dietary Zn addition reduced but waterborne Zn exposure increased hepatic lipid content. In contrast, muscle lipid content was reduced by waterborne Zn exposure but not by dietborne Zn exposure. The single exposure also affected several lipogenic enzymatic activities and expression of genes (in this article gene expression is taken synonymous to mRNA expression) related to lipogenesis and lipolysis. Pearson correlations among lipid content, enzymatic activities and mRNA expression levels were also observed, suggesting that changes at molecular and enzymatic levels may underlie the patterns of lipid metabolism and accordingly affect lipid deposition. For the first time, our study demonstrates the differential effect of different Zn exposure pathways on lipid metabolism at the molecular level in fish, indicating that the exposure route is critical to lipid deposition and metabolism.

Differential effects of acute and chronic zinc exposure on lipid metabolism in three extrahepatic tissues of juvenile yellow catfish Pelteobagrus fulvidraco

The aim of this study was to determine the potential mechanisms of exposure to waterborne zinc (Zn) on lipid metabolism in three extrahepatic tissues (ovary, muscle and mesenteric adipose tissue) of female yellow catfish Pelteobagrus fulvidraco. Female yellow catfish were chronically exposed to Zn (0.05, 0.35 or 0.86 mg Zn/l; duration of treatment 8 weeks) or acutely exposed to a high level of Zn (4.71 mg Zn/l for 96 h). Following the respective treatment, lipid deposition and mRNA levels of 11 genes (CPT IA, CPT IB, PPARα, PPARγ, SREBP-1, G6PD, 6PGD, FAS, ACCa, ACCb and LPL) involved in lipid metabolism were determined. Waterborne Zn exposure significantly reduced growth performance and lipid content in muscle but had no significant effect on lipid content in ovary and mesenteric adipose tissue. The change in the levels of the mRNA genes under study was Zn concentration-dependent and tissue-dependent. Pearson correlations between the mRNA levels of three transcriptional factors and enzymes in these tissues revealed that variations in gene expression as a result of the different Zn treatments underlay the patterns of lipid metabolism, which in turn affected fat storage and mobilization. To our knowledge, this is the first study to demonstrate the effect of waterborne Zn exposure on lipid metabolism in extrahepatic tissues at the molecular level. These results therefore contribute to our understanding of Zn-induced toxicity in fish.

Differential effects of the chronic and acute zinc exposure on carnitine composition, kinetics of carnitine palmitoyltransferases I (CPT I) and mRNA levels of CPT I isoforms in yellow catfish Pelteobagrus fulvidraco

The present study is conducted to determine the effect of acute and chronic zinc (Zn) exposure on carnitine concentration, carnitine palmitoyltransferases I (CPT I) kinetics, and expression levels of CPT I isoforms in liver, muscle and heart of yellow catfish Pelteobagrus fulvidraco. To this end, yellow catfish are subjected to chronic waterborne Zn exposure (0.05mgZnL−1, 0.35mgZnL−1 and 0.86mgZnL−1, respectively) for 8weeks and acute Zn exposure (0.05mgZnL−1 and 4.71mgL−1Zn, respectively) for 96h, respectively. Reduced Michaelis–Menten constants (Km) and maximal reaction rates (Vmax) values in liver and muscle are observed in fish exposed to chronic Zn concentration. In contrast, Vmax and Km values in heart increase with increasing Zn concentration. Chronic Zn exposure also significantly influences the contents of free carnitine (FC), total carnitine (TC) and acylcarnitine (AC) in liver and heart, but not in muscle. The acute Zn exposure significantly increases FC, AC, TC contents in liver and muscle, but reduces their contents in heart. The chronic and acute Zn exposure influences the mRNA levels of four CPT I isoforms (CPT Iα1b, CPT Iβ, CPT Iα2a and CPT Iα1a) in liver, muscle and heart. Furthermore, correlations are observed in the mRNA levels between CPT I isoforms and Km, and between isoforms expression and activity of CPT I. Thus, chronic and acute Zn exposure shows differential effects on carnitine content, CPT I kinetics and mRNA levels of four CPT I isoforms in yellow catfish, which provides new mechanism for Zn exposure on lipid metabolism and also novel insights into Zn toxicity in fish.

Differential effects of acute and chronic zinc (Zn) exposure on hepatic lipid deposition and metabolism in yellow catfish Pelteobagrus fulvidraco

The present study is conducted to determine the potential mechanisms of Zn on hepatic lipid deposition and metabolism for yellow catfish Pelteobagrus fulvidraco with 8-week chronic exposure to low Zn levels (Zn levels: 0.05, 0.35 and 0.86mg/l Zn, respectively) and 96-h acute exposure to a high Zn level (Zn level: 4.71mg/l Zn, respectively). For that purpose, hepatic lipid deposition and Zn accumulation, hepatic carnitine palmitoyltransferase I (CPT I) and lipoprotein lipase (LPL) activities, and the hepatic mRNA expression of ten genes involved in lipid metabolism are determined. Chronic (8 weeks) exposure to low Zn levels apparently increases hepatic lipid content, hepatosomatic index (HSI) (P<0.05) and LPL activity, and reduces hepatic CPT I activity. In contrast, the acute (96h) exposure to high Zn level reduces hepatic lipid content, HSI and LPL activity, and increases CPT I activity. The change of mRNA levels of genes related to lipid metabolism is Zn concentration-dependent. Pearson correlations among mRNA expression levels, lipid content, CPT I and LPL activities in liver are also observed in yellow catfish with the 8-week chronic Zn exposure. For the first time, our study demonstrates the effect of waterborne Zn exposure on lipid metabolism at the molecular levels in fish, which may contribute to understanding the mechanism of Zn-induced hepatic toxicity in fish.

Waterborne versus Dietary Zinc Accumulation and Toxicity in Daphnia magna: a Synchrotron Radiation Based X-ray Fluorescence Imaging Approach

Recent studies have suggested that exposure of the freshwater invertebrate Daphnia magna to dietary Zn may selectively affect reproduction without an associated increase of whole body bioaccumulation of Zn. The aim of the current research was therefore to investigate the hypothesis that dietary Zn toxicity is the result of selective accumulation in tissues that are directly involved in reproduction. Since under field conditions simultaneous exposure to both waterborne and dietary Zn is likely to occur, it was also tested if accumulation and toxicity under combined waterborne and dietary Zn exposure is the result of interactive effects. To this purpose, D. magna was exposed during a 16-day reproduction assay to Zn following a 5 × 2 factorial design, comprising five waterborne concentrations (12, 65, 137, 207, and 281 μg Zn/L) and two dietary Zn levels (49.6 and 495.9 μg Zn/g dry wt.). Tissue-specific Zn distribution was quantified by synchrotron radiation based confocal X-ray fluorescence (XRF). It was observed that the occurrence of reproductive inhibition due to increasing waterborne Zn exposure (from 65 μg/L to 281 μg/L) was accompanied by a relative increase of the Zn burdens which was similar in all tissues considered (i.e., the carapax, eggs, thoracic appendages with gills and the cluster comprising gut epithelium, storage cells and ovaries). In contrast, the impairment of reproduction during dietary Zn exposure was accompanied by a clearly discernible Zn accumulation in the eggs only (at 65 μg/L of waterborne Zn). During simultaneous exposure, bioaccumulation and toxicity were the result of interaction, which implies that the tissue-specific bioaccumulation and toxicity following dietary Zn exposure are dependent on the Zn concentration in the water. Our findings emphasize that (i) effects of dietary Zn exposure should preferably not be investigated in isolation from waterborne Zn exposure, and that (ii) XRF enabled us to provide possible links between tissue-specific bioaccumulation and reproductive effects of Zn.

Effect of waterborne zinc exposure on metal accumulation, enzymatic activities and histology of Synechogobius hasta

The present study was conducted to determine the metal accumulation, antioxidant enzymatic response, hepatic intermediary metabolism and histological changes in Synechogobius hasta exposed to 0.35 (control), 9.7 and 19.2 mg/L Zn, respectively, on the 0, 4th, 8th and 12th day. Waterborne Zn exposure significantly reduced hepatosomatic index, hepatic lipid contents and fatty liver occurrence rate, increased Zn, Fe and Mn contents and reduced the contents of Cu and Ca in liver, and increased muscle Zn content. Waterborne Zn exposure also significantly influenced enzymatic activities involved in antioxidant responses (superoxide dismutase, catalase, glutathione-S-transferase, malondialdehyde) in liver and spleen, and changed hepatic intermediary enzymatic activities (succinate dehydrogenase, malic dehydrogenase, lactate dehydrogenase, lipoprotein lipase, hepatic lipase), impaired the histological structure of the gill and spleen, and reduced vacuolated hepatocytes. Thus, our study demonstrated for the first time that waterborne Zn exposure could reduce fatty liver syndrome in S. hasta.

Probabilistic risk assessment of abalone Haliotis diversicolor supertexta exposed to waterborne zinc

This paper describes a risk assessment approach that integrates predicted tissue concentrations of zinc (Zn) with a concentration–response relationship and leads to predictions of survival risk for pond abalone Haliotis diversicolor supertexta as well as to the uncertainties associated with these predictions. The models implemented include a probabilistic bioaccumulation model, which linking biokinetic and consumer-resource models, accounts for Zn exposure profile and a modified Hill model for reconstructing a dose–response profile for abalone exposed to waterborne Zn. The growth risk is assessed by hazard quotients characterized by measured water level and chronic no-observed effect concentration. Our risk analyses for H. diversicolor supertexta reared near Toucheng, Kouhu, and Anping, respectively, in north, central, and south Taiwan region indicate a relatively low likelihood that survival is being affected by waterborne Zn. Expected risks of mortality for abalone were estimated as 0.46 (Toucheng), 0.36 (Kouhu), and 0.29 (Anping). The predicted 90th-percentiles of hazard quotient for potential growth risk were estimated as 1.94 (Toucheng), 0.47 (Kouhu), and 0.51 (Anping). These findings indicate that waterborne Zn exposure poses no significant risk to pond abalone in Kouhu and Anping, yet a relative high growth risk in Toucheng is alarming. Because of a scarcity of toxicity and exposure data, the probabilistic risk assessment was based on very conservative assumptions.