Metallothionein Concentrations Research Articles

Multi-model inference analysis of toxicological responses and levels of heavy metals in soft tissue of land snail Cornu aspersum caged in proximity to an industrial setting

The use of indicator species has become a standard monitoring approach in environmental risk assessment; however, pollutant bioavailability and its geographical distribution as well as species-sensitiveness could be affected by different factors. The present study investigated the potential use of the land snail Cornu aspersum as bioindicator of industrial air pollution by using cytological and biochemical responses and trace element levels in individuals caged in proximity to an industrial setting. Eleven sites were selected based on wind direction and distance. Snails were placed in holed plastic cages, transplanted to each site and maintained for 30 days fed at libitum and under constant humidity. Several oxidative stress responses including catalase (CAT) and glutathione-s-transferase (GST) activities, levels of malondialdehyde (MDA) and total metallothioneins (MTs) content were determined in snail hepatopancreas while lysosomal membrane stability (LMS) and loss of DNA integrity through micronuclei frequency (MN) were assessed in haemocytes. Trace elements were also analysed in snail whole soft tissues. Multi-model inference used to predict snail’s biological responses in relation to trace elements levels and distance from the industrial setting was able to disentangle the relative contributions of different influencing predictors and could be successfully applied in environmental risk assessment by using land snail as bioindicator species.

Biomarker responses in African sharptooth catfish, Clarias gariepinus (Burchell, 1822), as indicators of potential metal and organic pollution along the Vaal River system, South Africa

The Vaal River is of considerable economic and environmental importance. It flows through five provinces within South Africa and it is subject to numerous recreational and industrial activities, which often prove detrimental to the health of the system. To determine the extent of the impact that these anthropogenic stressors have on the river system, physiological effects in Clarias gariepinus were assessed during a 2016/2017 sample survey using a suite of biomarkers (exposure and effect). Inhibition of acetylcholinesterase and increased metallothionein concentrations were recorded in response to the increase in organic pollutants and metals. Oxidative stress biomarkers, such as superoxide dismutase, showed significant increases in response to possible recent pesticide exposures, whereas lipid peroxidation and protein damage were evident with increased malondialdehyde and protein carbonyl concentrations respectively. The data provide a novel account of the use of biomarkers to assess negative anthropogenic effects observed along the entire length of the Vaal River system.

Detection of Metallothionein Proteins by Enzyme-Linked Immunosorbent Assay (ELISA).

Metallothioneins (MTs) are low-molecular-weight, cysteine-rich proteins that bind to heavy metals. MTs play a key role in the homeostasis of metal ions, maintaining intracellular redox equilibria and free radical scavenging. In several studies, under different conditions such as cancer development, drug therapy and heavy metal stress, the unique structural changes and functional effects of MT were studied. Although several assays are available to monitor the content and type of Metallothionein (MT) from environmental samples or in biomedical assays, Enzyme-Linked Immunosorbent Assays (ELISA) became the preferred method of MT detection. ELISA is low in cost, specific, simple, and efficient. This review evaluates the advantages and disadvantages of using different types of ELISA in the detection of metallothioneins from environmental or clinical samples as well as ways of its validation and cross-validation.

Cigarette Smoking during Pregnancy: Effects on Antioxidant Enzymes, Metallothionein and Trace Elements in Mother-Newborn Pairs.

The effect of maternal smoking as a source of exposure to toxic metals Cd and Pb on superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity, metallothionein (MT), Cd, Pb, Cu, Fe, Mn, Se and Zn concentrations were assessed in maternal and umbilical cord blood and placenta in 74 healthy mother-newborn pairs after term delivery. Sparse discriminant analysis (SDA) was used to identify elements with the strongest impact on the SOD, GPx and MT in the measured compartments, which was then quantified by multiple regression analysis. SOD activity was lower in maternal and cord plasma, and higher in the placenta of smokers compared to non-smokers, whereas GPx activity and MT concentration did not differ between the groups. Although active smoking during pregnancy contributed to higher maternal Cd and Pb concentrations, its contribution to the variability of SOD, GPx or MT after control for other elements identified by SDA was not significant. However, an impaired balance in the antioxidant defence observed in the conditions of relatively low-to-moderate exposure levels to Cd and Pb could contribute to an increased susceptibility of offspring to oxidative stress and risk of disease development later in life. Further study on a larger number of subjects will help to better understand complex interactions between exposure to toxic elements and oxidative stress related to maternal cigarette smoking.

Sensitive and selective sensing system of metallothioneins based on carbon quantum dots and gold nanoparticles

In this study, we developed a sensitive and selective sensing system for the detection of metallothioneins (MTs). The system is based on the fluorescence resonance energy transfer (FRET) between carbon quantum dots (CQDs) and gold nanoparticles (AuNPs). In this method, the fluorescence emission of CQDs was quenched by AuNPs due to FRET. When MTs were added to the CQD–AuNP system, the strong combination between thiol group and Au made the CQDs release from AuNPs, and the fluorescence of CQDs was recovered. The CQD–AuNP system can detect the MTs in aqueous solution (pH 3.0, citrate–HCl buffer) selectively and sensitively with a short response time (15 min). Results show that the fluorescence recovery efficiency has a good linear relationship with the MTs concentration in the range of 12–210 nmol L−1, and the limit of detection was 5.25 × 10−9 mol L−1. Furthermore, the sensing system was utilized to determine MTs in human urine samples with satisfactory results. The proposed system exhibits the advantages of high sensitivity, high selectivity, easy operation and most importantly, low cost and non-cytotoxicity to detect protein MTs.

Mercury Exposure Assessment in Mother-Infant Pairs from Continental and Coastal Croatia.

The main source of mercury (Hg) exposure in the general population is fish. Another possible source is dental amalgam. Here, we compare the levels of Hg and selenium (Se) in samples of maternal and fetal origin collected shortly after childbirth of healthy postpartum women in the coastal (n = 96) and continental (n = 185) areas of Croatia related to maternal seafood/fish consumption. We also evaluated Hg concentrations and maternal serum metallothionein (MT2) concentrations in relation to the number of dental amalgam fillings, and MT2A-5A/G (rs28366003) polymorphism. The levels of Hg and Se in maternal hair and blood/serum, placenta and cord blood/serum increased in relation to increasing fish consumption with the highest values in subjects from the coast. The concentrations of each element and between elements correlated across the matrices. Increasing amalgam number correlated linearly with increased Hg levels in maternal and cord serum and was not associated with serum MT2. No association of MT2A-5A/G polymorphism and Hg or Se levels were found. The results confirmed higher fish consumption in coastal vs. continental Croatia and increases of both Hg and Se related to fish consumption in all analyzed samples. Increased blood Hg reflected the predominant MeHg share from seafood, while increased serum Hg matched exposure from dental amalgams.

Cytosolic and Metallothionein-Bound Hepatic Metals and Detoxification in a Sentinel Teleost, Dules auriga, from Southern Rio de Janeiro, Brazil.

Dules auriga, a native Brazilian teleost, was applied as a sentinel species regarding metal contamination at Ilha Grande Bay, previously considered a reference site in Southeastern Brazil. Cytosolic (S50) and metallothionein-bound (HTS50) hepatic iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), cadmium (Cd), and silver (Ag) were determined by inductively coupled plasma optical emission spectrometry (ICP-OES), while metallothionein (MT) concentrations were determined by polarography. Ag concentrations in both cytosolic fractions were below the limit of detection. All other HTS50 metal contents were significantly lower than S50 contents. No significant associations were found for MT. Fe and Mn S50 were positively and moderately correlated to total length, as well as HTS50 Mn, while total weight was correlated to both Mn fractions, suggesting that environmental Mn and Fe concentrations may influence fish growth. A moderate correlation between the condition factor and the S50 Cu fraction was observed, also indicating that Cu may affect fish growth. Inter-element correlations were observed, including between Cd, a toxic element, and Mn and Zn, both essential elements. Calculated molar ratios indicate that both Mn and Zn are in molar excesses compared with Cd, corroborating literature assessments regarding protective Mn and Zn effects against Cd. Lack of MT correlations suggests that metal concentrations may not be high enough to reach an MT induction threshold and that MT variability is probably linked to environmental metal concentrations. Therefore,the increased environmental contaminant levels observed in the study area indicate the need for biomonitoring efforts aiming at the application of efficient mitigation measures.

Biochemical Responses of the Bivalve Mollusk Unio tumidus Inhabiting a Small Power Plant Reservoir on the Dniester River Basin, Ukraine

Hydropower plants (HPPs) can affect the hydrological regime. However, biochemical responses of aquatic animals for the evaluation of this disturbing are not applied yet. The specimens of Unio tumidus were sampled in a reservoir (R) of a small HPP as well as downstream from the dam (DS). Biochemical indexes in the digestive gland and alkali labile phosphates (ALP) in the gonads were examined. The R-mollusks showed low cholinesterase, catalase and caspase-3 activities, and metallothionein concentration, but elevated levels of zinc and copper, oxidized glutathione and protein carbonyls. Concentrations of lactate, pyruvate and ALP, activity of superoxide dismutase and glutathione S-transferase, and lipid peroxidation level were similar in both groups. Integrated biomarker response (IBR/n) index (n = 13) was 2.17 and 0.29 in the R- and DS-groups correspondingly. We suggest that using integrative biological response based on the biochemical markers of bivalve mollusks can be a valid early warning step in assessing 'environmental flow' impact.

Microalgal Metallothioneins and Phytochelatins and Their Potential Use in Bioremediation.

The persistence of heavy metals (HMs) in the environment causes adverse effects to all living organisms; HMs accumulate along the food chain affecting different levels of biological organizations, from cells to tissues. HMs enter cells through transporter proteins and can bind to enzymes and nucleic acids interfering with their functioning. Strategies used by microalgae to minimize HM toxicity include the biosynthesis of metal-binding peptides that chelate metal cations inhibiting their activity. Metal-binding peptides include genetically encoded metallothioneins (MTs) and enzymatically produced phytochelatins (PCs). A number of techniques, including genetic engineering, focus on increasing the biosynthesis of MTs and PCs in microalgae. The present review reports the current knowledge on microalgal MTs and PCs and describes the state of art of their use for HM bioremediation and other putative biotechnological applications, also emphasizing on techniques aimed at increasing the cellular concentrations of MTs and PCs. In spite of the broad metabolic and chemical diversity of microalgae that are currently receiving increasing attention by biotechnological research, knowledge on MTs and PCs from these organisms is still limited to date.

Possible Mechanisms of Liver Injury Induced by Cadmium Sulfide Nanoparticles in Rat.

Cadmium is primarily utilized in the construction of particles known as quantum dots. Hepatotoxicity caused by microparticles of cadmium is very well known; however, toxicity of nanoparticles of cadmium is not well understood. The present study describes the toxicity of cadmium sulfide nanoparticles (CdSNPs) in the liver of rat. Adult Wistar rats were administered CdSNPs (10mg/kg) on alternate days for 45days. Serum enzymes (ALT, AST, ALP), biomarkers of lipid peroxidation (MDA, H2O2, and NO), and metallothionein concentration were determined. Histopathological and TEM observations were also made to record morphological changes. CdSNPs (10mg/kg) induced significant changes in the structure and function of liver. Values of serum enzymes and reactive species increased significantly in rats treated with CdSNPs in comparison to CdS-treated rats. Histopathological observations showed extensive parenchymal degeneration. Ultrastructural studies exhibited proliferation of endoplasmic reticulum, microsomes, and lysosomes. It is concluded that NP-membrane interaction leads to the generation of reactive species that alter membrane integrity and induce oxidative stress. These events may activate cell death pathways in hepatocytes.

Banded tetra (Astyanax aeneus) as bioindicator of trace metals in aquatic ecosystems of the Yucatan Peninsula, Mexico: Experimental biomarkers validation and wild populations biomonitoring.

Bioindicator organisms are important tools in environmental monitoring studies. Understanding this, the overall goal of the present study was to evaluate the sensitivity and viability of the native fish species Banded tetra, (Astyanax aeneus; Günther, 1860), widely spread in the aquatic ecosystems of the Yucatan Peninsula in Mexico, as a bioindicator organism. In order to do this, we performed a bioassay at sublethal concentrations using copper (CuSO4) to experimentally evaluate and validate the relationship between the trace metals and oxidative stress biomarkers response [(catalase (CAT), lipoperoxidation content (LPO)], detoxification [(glutathione S-transferase (GST), metallothionein content (MT)] and neurotoxicity (AChE) in muscle of A. aeneus. Results showed changes in biomarkers after 96h: Catalase activity (CAT) was significantly higher above 1.5 and 2mg/L (154.35 and 172.50% increase, respectively); lipid peroxidation contents (TBARS), GST activity, and MT content were very similar to CAT activity at 1.5 and 2mg/L of Cu. In terms of neurotoxicity, AChE activity was significantly inhibited at 0.1mg/L (64%; p<0.001) and 0.5mg/L (44%; p<0.001) of Cu. Based on the bioassay results, we performed a trace metal monitoring campaign in muscle of A. aeneus caught in 15 sites with different anthropogenic activities, during the summer of 2017, to establish a baseline of trace metals pollution in the state of Campeche. A. aeneus showed the highest trace metal accumulation in the following order: Al>Fe>Mn>Zn>Cu>Hg>Cr>Pb>Cd>V>As, while sites were arrange as follows: Xnoha lagoon>Palizada River>Candelaria River>Ululmal>Maravillas>López Mateos. PCA showed a cluster between biomarkers (GST, CAT, TBARS, and MT) and concentration of metals (Cd, Cu, Fe, Zn, Hg and Cr). Conversely, AChE inhibition was not related to a specific metal, but highest inhibitions (>50%) were present in those sites with intensive agricultural practices. These results determined that, based on its physiological response and trace metal bioaccumulation, Astyanax aeneus can be considered a good bioindicator for evaluating the presence of trace metals in tropical aquatic systems of the Yucatan Peninsula.

Mitigation of zinc toxicity through differential strategies in two species of the cyanobacterium Anabaena isolated from zinc polluted paddy field

The present study describes the physiological and biochemical mechanisms of zinc tolerance in two heterocytous cyanobacteria i.e. Anabaena doliolum and Anabaena oryzae, treated with their respective LC50 concentrations of zinc (3 and 4.5 mg L−1) for eight days. The feedbacks were examined in terms of growth, metabolism, zinc exclusion, zinc accumulation, oxidative stress, antioxidants and metallothionein contents. Although the growth and metabolic activities were reduced in both the cyanobacterium, maximum adversity was noticed in A. doliolum. The higher order of abnormalities in A. doliolum was attributed to excessive accumulation of zinc and enhanced reactive oxygen species (ROS) production. However, the comparatively higher growth and metabolic activities of A. oryzae were ascribed to the lower accumulation of zinc as a result of released polysaccharides mediated zinc exclusion, synthesis of zinc chelating metallothioneins and subsequent less production of ROS. The oxidative stress and macromolecular damages were prominent in both the cyanobacterium but the condition was much harsher in A. doliolum which may be explained by its comparatively low antioxidative enzyme activities (SOD, APX and GR) and smaller amount of ascorbate-glutathione-tocopherol contents than that of A. oryzae. However, sustenance of 50% growth by A. doliolum under zinc stress despite severe cellular damages was attributed to the enhanced synthesis of phenolics, flavonoids, and proline. Thus, differential zinc tolerance in A. doliolum and A. oryzae is possibly the outcome of their distinct mitigation strategies. Although the two test organisms followed pseudo second order kinetics model during zinc biosorption yet they exhibited differential zinc biosorption capacity. The cyanobacterium A. oryzae was found to be more efficient in removing zinc as compared to A. doliolum and this efficiency makes A. oryzae a promising candidate for the phycoremediation of zinc polluted environments.

Exposure to Dichlorodiphenyldichloroethylene (DDE) and Metallothionein Levels in Rats Fed with Normocaloric or High-Fat Diet: A Review

The growing number of studies on metallothioneins (MTs), cysteine-rich metal-binding proteins, have been disclosing new functions of these proteins. Thanks to their inducibility, they were considered to play a pivotal role in regulating trace metals homeostasis and in detoxification from heavy metals; nowadays, it is known that they are involved in various physiological and pathological processes, such as regulation of apoptosis, elimination of free radicals, and protection of nucleic acids against toxic insults. MT induction has been demonstrated following stress factors other than heavy metals, such as endocrine-disrupting chemicals, insecticides, and herbicides. However, retrieved data are often controversial: in some cases, xenobiotics elicit MT expression and synthesis; under different conditions, they lead to a decrease in cellular MT content. This review describes the MT response to dichlorodiphenyltrichloroethane (DDT) contamination in mammalian tissues. In particular, attention focuses on changes in MT expression, synthesis, and localization in rat liver, kidneys, and testes following oral administration of dichlorodiphenyldichloroethylene (DDE), the main metabolite of DDT, under normal dietary conditions or in combination with a high fat diet potentially able to increase the cellular uptake of this lipophilic pesticide. The potential connection between MT expression and synthesis, lipophilic substances and trace metals availability is also discussed.

Metallothionein 2A gene polymorphisms in relation to diseases and trace element levels in humans.

Human metallothioneins are a superfamily of low molecular weight intracellular proteins, whose synthesis can be induced by essential elements (primarily Zn and Cu), toxic elements and chemical agents, and stress-producing conditions. Of the four known isoforms in the human body MT2 is the most common. The expression of metallothioneins is encoded by a multigene family of linked genes and can be influenced by single nucleotide polymorphisms (SNPs) in these genes. To date, 24 SNPs in the MT2A gene have been identified with the incidence of about 1 % in various population groups, and three of them were shown to affect physiological and pathophysiological processes. This review summarises current knowledge about these three SNPs in the MT2A gene and their associations with element concentrations in the body of healthy and diseased persons. The most investigated SNP is rs28366003 (MT2A −5 A/G). Reports associate it with longevity, cancer (breast, prostate, laryngeal, and in paranasal sinuses), and chronic renal disease. The second most investigated SNP, rs10636 (MT2A +838G/C), is associated with breast cancer, cardiovascular disease, and type 2 diabetes. Both are also associated with several metal/metalloid concentrations in the organism. The third SNP, rs1610216 (MT2A −209A/G), has been studied for association with type 2 diabetes, cardiomyopathy, hyperglycaemia, and Zn concentrations. Metallothionein concentrations and MT2A polymorphisms have a potential to be used as biomarkers of metal exposure and clinical markers of a number of chronic diseases. This potential needs to be studied and verified in a large number of well-defined groups of participants (several hundreds and thousands) with a focus on particular physiological or pathological condition and taking into consideration other contributing factors, such as environmental exposure and individual genetic and epigenetic makeup.

Ecological Monitoring of the Unal Depression, Northern Ossetiya–Alania, Using Techniques of Biogeochemical Indication

The paper presents analysis of the accumulation of toxic and vital chemical elements in discrete links of the food chain under conditions of a biogeochemical province in relation to the accumulation of phytochelatins and metallothioneins in the leaves of white birch (Betula pubescens). Gradients of metal concentrations are determined in the soils with increasing distance from the Unal tailing dump and the effects of these gradients on the concentrations of the elements in birch leaves. Correlations are determined between the concentrations of toxic elements in the pulp and leaves separately for the native sample and water washout. A strong correlation was found between the total dissolved concentration of all metals and the total metallothionein and glutathione concentrations. It is hypothesized that the biochemical system of glutathione is more closely related to the accumulation of metals by plants than the system of metallothioneins. Estimates are made whether it is possible to apply the method of bioindication of the ecological state of a territory by determining the fluctuating asymmetry of birch leaves. This bioindication parameter was evaluated separately for each of the cardinal points. It is demonstrated that a high total concentrations of the metals in soils and birch leaves is correlated with an increase in the fluctuating asymmetry of the leaves and the concentration level of sulfur-bearing biological active compounds. The bioindication technique is however still far from perfect and is difficult to apply and, hence, requires further studies.

Total and subcellular Ti distribution and detoxification processes in Pontoporia blainvillei and Steno bredanensis dolphins from Southeastern Brazil

Titanium (Ti), used in many dailyuse products, such as shampoos and sunscreen filters, in the form of TiO2 nanoparticles (NPs), may elicit adverse marine biota effects. Marine mammal Ti data is scarce, and subcellular distribution and detoxification information is non-existent. Ti concentrations and metalloprotein detoxification in Pontoporia blainvillei and Steno bredanensis dolphins from Southeastern Brazil were assessed. Metallothionein (MT) concentrations were determined spectrophotometrically, total and subcellular Ti, by ICP-MS and detoxification, by HPLC-ICP-MS. Ti detoxification occurred through MT complexation. Statistical Ti-MT associations were observed in S. bredanensis liver, indicating TiO2 NPs contamination, as Ti binds to MT only as NPs. MT-Ti correlations were observed for both the coastal (P. blainvillei) and offshore (S. bredanensis) dolphins, evidencing oceanic TiO2 diffusion. Ti detoxification through binding to reduced glutathione occurred in both species. Thermostable subcellular fractions are a valuable tool for cetacean Ti detoxification assessments and should be applied to conservation efforts.

Metallothionein level, non-specific esterases, fitness-related traits and integrated biomarker response (IBR) in larvae of Lymantria dispar L. (Lepidoptera) originating from unpolluted and polluted locations after chronic cadmium treatment

Long-term exposure of invertebrate populations to pollution might result in adaptations that must be taken into account when evaluating physiological parameters as biomarkers of contamination. In the present study we investigated whether previous exposure of gypsy moth, Lymantria dispar L. (Lepidoptera), population to pollution affects the sensitivity of larvae to cadmium treatment under laboratory conditions. Our goals in the current study were to compare metallothionein (MT) level and non-specific esterase activity including isoform expression in the midgut and fitness-related traits between caterpillars originating from polluted and unpolluted localities after chronic oral intake (50 and 100 µg Cd/g dry food). Besides being individually assessed as biomarkers of cadmium contamination, the responses of above parameters related to the midgut (MT concentration, specific activity of esterases and midgut mass) were summarized into the integrated biomarker response (IBR). Upon cadmium exposure, MT concentration increased significantly only in larvae originating from the polluted site, whereas non-specific esterase activity decreased in caterpillars from the unpolluted forest. We noticed similar patterns of esterase isoforms in both populations, including isoform number three that appeared only after exposure to the lower cadmium concentration. Midgut mass was reduced only in the group from the unpolluted locality exposed to the higher cadmium concentration. IBR index increased in a concentration-dependent manner in both populations, reflecting higher sensitivity of larvae from the unpolluted locality to the metal. Neither cadmium exposure nor population origin influenced larval survival. While MT concentration could be applied as a biomarker of cadmium exposure in populations that have been previously exposed to pollution for a longer time, non-specific esterase activity could indicate metal presence at unpolluted locations. IBR index and expression of esterase isoform number three could be used universally, regardless of the exposure history, except the latter might be limited to lower cadmium concentrations.

The effect of Roundup on the bivalve Unio tumidus mollusk utilizing ex vivo approach

Glyphosate is the worldwide used herbicide of the first priority. However, its biochemical effects in the aquatic animals are studied scantly. The ex vivo approach has been recently proposed to provide the express evaluation of the adverse impact without the treating of the organisms. The aim of this study was to verify this approach for the assessment of the toxicity of glyphosate to the bivalve mollusk. The samples of the gills and digestive gland tissues of freshwater bivalve Unio tumidus mollusk were exposed to a range of the concentrations of glyphosate (commercial formulation Roundup MAX) at the concentrations 13.3, 26.7, 66.8 and 133.6 µg×L -1 during 2 h at 20 °C followed by 15 h at ~ 2–4 o C. The markers of oxidative injury (total antioxidant activity, end-products of lipid peroxidation (TBARS) and protein carbonyls (PC)), cellular low weight thiols GSH/GSSG and metallothionein (MT), and cholinesterase activity as the index of neurotoxicity were analyzed. We also assayed the index of cell vitality as the lysosomal membrane stability from the Neutral Red Retention (NRR) test. The results have shown that the lowest concentrations of glyphosate caused the most prominent changes of the indices: the decrease of MT concentration (by ~ two times) and cholinesterase activity. The total antioxidant activity was decreased substantially in all exposures correspon­dingly to a decrease of the MT and/or GSH concentrations. However, the levels of TBARS and PC were not changed comparing to control detecting the early stage of the injury. Surprisingly, NRR increased in the exposures to higher concentrations of glyphosate, probably due to strong chelating ability of glyphosate or other compounds of formulation. This study allows us to detect the earlier biological effects of glyphosate in the low environmentally realistic concentrations. Further validation of this approach needs the comparison of the results in the ex vivo and in vivo experiments. Keywords: Roundup; bivalve mollusk; cytotoxicity; ex vivo

Effects of Dietary Zinc on Performance, Zinc Transporters Expression, and Immune Response of Aged Laying Hens.

This study was to investigate the effects of dietary zinc (Zn) supplementation on performance, zinc transporter gene expression, and immune function in aged laying hens. In experiment 1, twenty 31-week-old hens (young) and twenty 60-week-old hens (old) with the same genetic background were fed with the same diet for 4 weeks. In experiment 2, a basal diet supplemented with zinc sulfate (ZnS) and zinc glycine chelate (ZnG) at 30, 60, 90, and 120 mg Zn/kg to constitute nine experimental diets. Eight hundred and ten 60-week-old layers were distributed in a completely randomized experimental design with 9 treatments, 6 replicates of 15 birds each, and birds were fed for 10 weeks. In experiment 1, results showed that zinc and metallothionein (MT) concentration in the shell gland of old hens was significantly lower than young layers (P < 0.05). Zinc transporters ZnT1, 4, 5, 6, and 7 messenger RNA (mRNA) abundance in old layers were significantly lower versus the young (P < 0.05). In experiment 2, results indicated that dietary zinc supplementation did not significantly affect the laying rate, average feed intake, egg weight, feed conversion efficiency, broken egg rate, or mortality (P > 0.05). Supplemental ZnG significantly improved eggshell breaking strength than ZnS, with a higher alkaline phosphatase (ALP) activity and more abundant ZnT4 expression in shell gland versus ZnS (P < 0.05). ZnG supplementation at 90 mg Zn/kg affected the duodenal mucus by significantly increasing ZnT1, 6, 7, ZIP13, and MT-4 mRNA level (P < 0.05). Zinc level significantly increased bovine serum albumin (BSA) antibody concentrationon 14 day after BSA injection (P < 0.05). Supplementation of ZnG improved eggshell quality of aged layers by upgrading zinc transporter expression in the shell gland and intestine also enhanced humoral immunity.

Effects of sulfur on the toxicity of cadmium to Folsomia candida in red earth and paddy soil in southern Fujian

Sulfur has been shown to mitigate the toxic effects of metals on soil organisms. Here we report the effects of sulfur on cadmium toxicity to the collembolan Folsomia candida in soil, including its effects on glutathione (GSH) level, catalase (CAT) activity and metallothionein (MT) content. Following sulfur treatment, catalase, glutathione and metallothionein activities were all significantly increased in cadmium-contaminated soil, and as the cadmium concentration increased, the activities decreased. In addition, because of the reducing effects of pH and organic matter on cadmium bioavailability, the bioavailable cadmium varied among soils of different pH values and organic matter contents, causing the catalase activity, glutathione content and metallothionein levels of F. candida to vary among soils. Our study suggests that sulfur can affect the toxicity of certain concentrations of cadmium and that soil properties are very important to consider. This study provides insight into the effects of sulfur application on soil animals.