Waterborne Zinc Exposure Research Articles

Induction of sex ratio change and intersex in Tegillarca granosa (Bivalvia: Arcidae) by waterborne zinc exposure

This study aims to investigate the effect of zinc on the sex ratio and intersexuality of Tegillarca granosa, a sequential hermaphroditic bivalve that undergoes sex change during the gonadal inactive stage. Groups of T. granosa in the gonadal inactive stage were first exposed to zinc under indoor aquarium conditions for 5 weeks, and then under wild conditions for 3 months. The proportion of males and females tended to increase and decrease, respectively as the zinc concentration increased. Intersexuality increased with an increase in zinc concentration, and occurred more in female than male groups in all zinc exposed groups. The intersex of T. granosa exposed to zinc could be divided into interfollicular and intrafollicular types according to the histological pattern. The results of this study suggest that zinc exposure higher than a certain concentration may induce male sexuality during the gonadal inactive stage in the same population of T. granosa, and could result in unequal sex ratios as well as intersex.

Regulation of metal homeostasis and zinc transporters in early-life stage zebrafish following sublethal waterborne zinc exposure

In the present research, the effects of exposure to a sublethal concentration of zinc (Zn) on metal and ion homeostasis, and the regulation and the localization of various Zn transporters (i.e., the Zrt-Irt Like Protein (ZIP) family of Zn transporters), were investigated in zebrafish (Danio rerio) during early development. Exposure to an elevated level of Zn [4 μM (high) vs. 0.25 μM (control)] from 0 day post-fertilization (dpf) resulted in a significant increase in the whole body content of Zn at 5 dpf. A transient decrease in the whole body calcium (Ca) level was observed in 3 dpf larvae exposed to high Zn. Similarly, whole body nickel (Ni) and copper (Cu) contents were also reduced in 3 dpf larvae exposed to high Zn. Importantly, the magnitude of reduction in whole body Ni and Cu contents following Zn exposure was markedly higher than that in Ca content, suggesting that internal Ni and Cu balance were likely more sensitive to Zn exposure in developing zebrafish. Exposure to high Zn altered the mRNA expression levels of specific zip transporters, with an increase in zip1 (at 3 dpf) and zip8 (at 5 dpf), and a decrease in zip4 (at 5 dpf). The expression levels of most zip transporters tended to decrease from 3 dpf to 5 dpf with the exception of zip4 and zip8. Results from in situ hybridization revealed that several zip transporters exhibited distinct spatial distribution (e.g., zip8 in the intestinal tract, zip14 in the pronephric tubules). Overall, our findings suggested that exposure to sublethal concentrations of Zn disrupts the homeostasis of essential metals during early development and that different ZIP transporters may play unique roles in regulating Zn homeostasis in various organs in developing zebrafish.

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.

Distribution and speciation of zinc in the gills of rainbow trout (Oncorhynchus mykiss) during acute waterborne zinc exposure: Interactions with cadmium or copper

We utilized micro X-ray fluorescence imaging (μ-XFI) and micro X-ray absorption near-edge spectroscopy (μ-XANES), which are both synchrotron-based techniques to investigate Zn distribution profile, its co-localization patterns with Ca, S, and Fe and speciation in the gills of rainbow trout (RBT). Fish (~100 g) were exposed to acutely toxic levels of waterborne Zn alone and in combination with waterborne Cd or Cu for 24 h (each at 1 × 96 h LC50). Gill sections were prepared and analyzed at the VESPERS beamline of the Canadian Light Source. The primary lamellae of the fish gill were found to be the primary area of Zn accumulation. These regions also correspond to the zones of mitochondria rich cells localization in fish gills, supporting the putative roles of these cells in metal uptake. Zn was also found to predominantly co-localize with Ca and S, but not with Fe, indicating the roles of Ca and S in intracellular Zn handling. Zn distribution in the gill was markedly reduced during co-exposure to Cd, but not to Cu, suggesting a competitive interaction between Zn and Cd for uptake. The speciation of Zn in the gill was dominated by Zn-phosphate, Zn-histidine and Zn-cysteine species; however, the interactions of Zn with Cd or Cu resulted in the loss of Zn-cysteine. Overall, our findings provide important novel insights into the interactions of Zn, Cd and Cu in the fish gill, which may ultimately help to explain the mechanisms underlying the acute toxicity of these metals in binary mixture to fish.

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.

Zinc Distribution in the Organs of Adult Fundulus heteroclitus After Waterborne Zinc Exposure in Freshwater and Saltwater

Zinc (Zn) is an essential micronutrient to aquatic organisms, but increased concentrations may result in accumulation and toxic effects. Water chemistry is known to influence the uptake of Zn in aquatic biota; therefore, organisms inhabiting environments with variable salinities may exhibit different patterns of Zn accumulation. Likewise, metal uptake can vary in fish as a consequence of ionoregulatory status (acclimated to freshwater or saltwater). The euryhaline fish, Fundulus heteroclitus, was exposed to a control and two increased Zn concentrations (15 and 75 μg/L in moderately hard freshwater and 100 and 1,000 μg/L in 35 g/L saltwater) for 7 days. The ionic Zn concentrations were equivalent in the 75 μg/L Zn treatment in freshwater and the 100 μg/L Zn treatment in saltwater. Throughout the 7-day experiments, fish were dissected, and organ Zn distribution was quantified in the gill, intestine, liver, gall bladder, heart, and carcass. Different patterns of Zn accumulation were observed in F. heteroclitus dependent on exposure medium. Despite lower exposure concentrations, F. heteroclitus accumulated more Zn in freshwater than in saltwater in all of the organs analyzed with the exception of the carcass. In addition, there were correlations between Zn distribution and known physiological mechanisms related to osmoregulation in F. heteroclitus. Furthermore, this research suggests that F. heteroclitus are more susceptible to Zn accumulation in freshwater environments.

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.

Interaction between dietary calcium supplementation and chronic waterborne zinc exposure in juvenile rainbow trout, Oncorhynchus mykiss

This study investigated the effects of dietary Ca 2+ on branchial Ca 2+ and Zn 2+ uptake, new and total zinc accumulation in target tissues (gill, liver and kidney), calcium and zinc homeostasis, and acute tolerance to waterborne zinc in fish chronically exposed to waterborne zinc. Juvenile rainbow trout ( Oncorhynchus mykiss) were maintained on a calcium-enriched diet [41.2 mg vs. 21.2 mg (control) calcium/g dry wt. of food] and chronic waterborne zinc exposure (2.3 μmol/L), both separately and in combination, for 28 days. Calcium-supplemented diet in the absence of waterborne zinc significantly reduced branchial Ca 2+ and Zn 2+ influx rates, and new and total zinc accumulations in target tissues relative to control. However it did not protect against the acute zinc challenge. In contrast, waterborne zinc exposure significantly increased branchial Ca 2+ and Zn 2+ influx rates, new and total zinc concentrations in target tissues, and acute zinc tolerance relative to control. Interestingly, no such changes in any of these parameters were recorded in fish treated simultaneously with elevated dietary Ca 2+ and waterborne zinc, except acute zinc tolerance which was highest among all the treatments. Thus, we conclude that the interactions between elevated dietary Ca 2+ and waterborne zinc can protect freshwater fish against waterborne zinc toxicity.

Costs of chronic waterborne zinc exposure and the consequences of zinc acclimation on the gill/zinc interactions of rainbow trout in hard and soft water

AbstractJuvenile rainbow trout were exposed to zinc in both moderately hard water (hardness = 120 mg CaCO3/L, pH = 8.0, Zn = 150 μg/L or 450 μg/L) and soft water (hardness = 20 mg CaCO3/L, pH = 7.2, Zn = 50 μg/L or 120 μg/L) for 30 d. Only the 450 μg/L zinc—exposed fish experienced significant mortality (24% in the first 2 d). Zinc exposure caused no effect on growth rate, but growth affected tissue zinc levels. Whole body zinc levels were elevated, but gills and liver showed no consistent increases relative to controls over the 30 d. Therefore, tissue zinc residues were not a good indicator of chronic zinc exposure. After the 30‐d exposure, physiological function tests were performed. Zinc was 5.4 times more toxic in soft water (control 96 h LC50s in hard and soft water were 869 μg/L and 162 μg/L, respectively). All zinc‐exposed trout had acclimated to the metal, as seen by an increase in the LC50 of 2.2 to 3.9 times over that seen in control fish. Physiological costs related to acclimation appeared to be few. Zinc exposure had no effect on whole body Ca2+ or Na+ levels, on resting or routine metabolic rates, or on fixed velocity sprint performance. However, critical swimming speed (UCrit) was significantly reduced in zinc‐exposed fish, an effect that persisted in zinc‐free water. Using radioisotopic techniques to distinguish new zinc incorporation, the gills were found to possess two zinc pools: a fast turnover pool (T1/2 = 3–4 h) and a slow turnover pool (T1/2 = days to months). The fast pool was much larger in soft water than in hard water, but at most it accounted for <3.5% of the zinc content of the gills. The size of the slow pool was unknown, but its loading rate was faster in soft water. Chronic zinc exposure was found to increase the size of the fast pool and to increase the loading rate of the slow pool.