Concentration of fifty-six elements in excreta of penguins from the Antarctic Peninsula area.

Seabird feces as indicators of the exposure to environmental contaminants have been studied worldwide. Penguins are indicator species for marine pollution, but their role as biovectors of rare earth elements (REEs) to ecosystems have been little studied. The present study quantified the concentration of REEs and trace elements (TEs) in feces of gentoo penguin (Pygoscelis papua). Adult penguin excreta from Fildes Bay (King George Island) and Yelcho Base (Palmer Archipelago) were collected and then analyzed by ICP-MS. Among REEs, levels ranged from 0.0038 to 1.02 µg g-1 d.w. for Lu and Ce, respectively. For TEs, the levels varied widely through the sample set, with Al, Fe, Sr, Zn and Ti as the highest mean levels, particularly at Fildes Bay. The data show that gentoo penguins act as a biovector organism by transporting TEs and REEs from the sea to land via excreta, reaffirming that this species acts as an important biovector organism in Antarctic ecosystems. The potential impacts of this process on Antarctic ecosystems needs further research.

Occurrence of rare earth elements (REEs) and trace elements (TEs) in feathers of adult and young Gentoo penguins from King George Island, Antarctica.

Penguins are sentinel species for marine pollution, but their role as potential biovectors of REEs or TEs to ecosystems has been poorly studied. The present study analyzed (ICP-MS) feathers of young and adult Gentoo penguins from Fildes Bay, for 63 elements (including 15 REEs). Most of the REEs were present at very low levels, ranging from 0.002 (Lu) to 0.452 (Sm) µg g-1 d.w., several orders of magnitude lower than TEs. The content of TEs varied widely, with Al, Fe, Zn, Sr, Ba, Ti and Mn as the seven having the highest concentrations in the feathers of both age groups. The results show that P. papua deposits REEs and TEs through the feathers on the penguin rockery, whose potential actual impacts and long-term fate in remote regions need deeper research. This work presents essential baseline data that will be useful for further studies on Antarctic penguins.

Trace and rare earth elements in excreta of two species of marine mammals from South Shetland Islands, Antarctica.

Pinnipeds are sentinel species for marine pollution, but their role as vectors of trace elements (TEs) or rare earth elements (REEs) to ecosystems has been poorly studied. The present study tested pinniped feces for 61 elements, including REEs. Feces of adult seals (Mirounga leonina, Hydrurga leptonyx) from Fildes Bay, King George Island, Antarctica, were analyzed by ICP-MS. TEs varied by several orders of magnitude across the suite examined herein, with Fe, Al, Zn, Mn, HgII and Sr as the top six in both species. Of the REEs, Ce, Dy, Er, Eu, Gd, Ho, La, Lu, Nd, Pr, Sc, Sm, Tb, Y and Yb were found consistently in all samples and ranged from 0.935 to 0.006 µg g-1 d.w. The results show that both species act as biovector organisms of TEs and REEs through feces in remote environments, whose actual impacts and long-term fate need further exploration.

Exposure of silver nanocolloids causes glycosylation disorders and embryonic deformities in medaka.

Silver nanomaterials such as silver nanocolloids (SNC) contribute to environmental pollution and have adverse ecological effects on aquatic organisms. In particular, chemical exposure of fish during embryogenesis leads to deformities and puts the population at risk. Although glycans and glycosylation are known to be important for proper morphology in embryogenesis, little glycobiology-based research has examined morphological disorders caused by environmental pollutants. This study addressed the glycobiological effects of SNC exposure on medaka embryogenesis. After exposure of medaka embryos to SNC, deformities such as small heads and deformed eyes were observed. The expression of five glycan-related genes (alg2, gnsb, b4galt2, b3gat1a, and b3gat2) was significantly altered, with changes depending on the embryonic stage at exposure, with more severe deformities with exposure at earlier stages. In situ hybridization analyses indicated that the five genes were expressed mainly in the head region; exposure of SNC suppressed alg2 and gnsb and enhanced b4galt2 and b3gat1a expression relative to controls on day 7. Loss (siRNA)- and gain (RNA overexpression)-of-function experiments confirmed that alg2, gnsb, and b4galt2 are essential for embryogenesis. The effects of SNC exposure on glycan synthesis were estimated by glycan structure analysis. In the medaka embryo, high mannose-type glycans were dominant, and SNC exposure altered glycan synthesis. The alteration was more significant when exposure occurred at an early stage of medaka embryogenesis. Thus, SNC exposure causes embryonic deformities in medaka embryos through disordered glycosylation.

Ecological Risks Due to Immunotoxicological Effects on Aquatic Organisms.

The immunotoxic effects of some anthropogenic pollutants on aquatic organisms are among the causes of concern over the presence of these pollutants in the marine environment. The immune system is part of an organism's biological defense necessarily for homeostasis. Thus, the immunotoxicological impacts on aquatic organisms are important to understand the effects of pollutant chemicals in the aquatic ecosystem. When aquatic organisms are exposed to pollutant chemicals with immunotoxicity, it results in poor health. In addition, aquatic organisms are exposed to pathogenic bacteria, viruses, parasites, and fungi. Exposure to pollutant chemicals has reportedly caused aquatic organisms to show various immunotoxic symptoms such as histological changes of lymphoid tissue, changes of immune functionality and the distribution of immune cells, and changes in the resistance of organisms to infection by pathogens. Alterations of immune systems by contaminants can therefore lead to the deaths of individual organisms, increase the general risk of infections by pathogens, and probably decrease the populations of some species. This review introduced the immunotoxicological impact of pollutant chemicals in aquatic organisms, including invertebrates, fish, amphibians, and marine mammals; described typical biomarkers used in aquatic immunotoxicological studies; and then, discussed the current issues on ecological risk assessment and how to address ecological risk assessment through immunotoxicology. Moreover, the usefulness of the population growth rate to estimate the immunotoxicological impact of pollution chemicals was proposed.

Structural analysis of N-glycans in medaka gut exposed to silver and titanium dioxide nanoparticles.

Nanomaterials are in general use in a broad range of industries. However, there are concerns that their intense use leads to heavy damage to the aquatic environment, and their discharge harms many aquatic organisms. N-Glycans are widely distributed in eukaryotic organisms and are intimately involved in most life phenomena. However, little is known about N-glycans in aquatic organisms exposed to nanomaterials. In this study, we investigated how nanomaterials affect N-glycans in the gut of adult female medaka. We found that silver nanoparticles exposure had little effect on gut N-glycans, whereas titanium dioxide nanoparticles (TiO2NPs) exposure increased the relative levels of several N-glycans in comparison with control. Structural analysis showed high levels of N-glycans of the high-mannose type, of which five N-glycans were free N-glycans with one ß-N-acetylglucosamine residue on the reducing end. The levels of free N-glycans are closely related to protein quality control in the endoplasmic reticulum and cytosol. Our results suggest that TiO2NPs exposure increases the levels of misfolded glycoproteins, resulting in generation of considerable amounts of free N-glycans. Our findings also suggest that TiO2NPs exposure suppresses cytosolic α-mannosidase trimming. This study provides new evidence for the effect of TiO2NPs on medaka gut from the aspect of environmental glycobiology.

Proinflammatory response caused by lead nanoparticles triggered by engulfed nanoparticles.

In this study, the cellular effects of lead (Pb) nanoparticles with a primary particle size of 80 nm were evaluated in two types of cell lines: human lung carcinoma A549 and macrophage-differentiated THP-1 cells (dTHP-1). The cellular responses induced by the Pb nanoparticles varied among the cell types. Exposure to Pb nanoparticles for 24 h at a concentration of 100 µg/ml induced interleukin-8 (IL-8) expression in dTHP-1 cells. Induction of IL-8 expression in A549 was lower than dTHP-1 cells. Pb nanoparticles also induced the gene expression of heme oxygenase-1 in dTHP-1 cells but not in A549 cells. Though cellular uptake of Pb nanoparticles was observed in both the cell types, the amount of internalized Pb particles was lower in A549 cells than that in dTHP-1 cells. Gene expression of metallothionein 2A was remarkably enhanced by Pb nanoparticle exposure in dTHP-1 cells. Compared with Pb nanoparticles, induction of cytokines caused by lead nitrate (Pb[NO3 ]2 ), a water-soluble Pb compound, was smaller. In conclusion, the present study revealed that Pb nanoparticles induced a stronger cellular response than Pb(NO3 )2 , primarily by eliciting cytokine production, in a cell type-dependent manner.

Do Polystyrene Beads Contribute to Accumulation of Methylmercury in Oysters?

To clarify whether microplastics contribute to elevated bioaccumulation of methylmercury (MeHg) in aquatic organisms, we studied the sorption pattern of MeHg on polystyrene beads (PBs) and evaluated MeHg accumulation, via uptake of MeHg-adsorbed PB, in the oyster Crassostrea gigas. MeHg-cysteine conjugates were added to seawater at 10, 100, and 1000 µg/L as Hg. Polystyrene beads (φ = 0.02, 0.2, and 2 µm) were immersed in the seawater for 24 h. The concentrations of total mercury (T-Hg) adsorbed onto the PBs were then measured using the reduction vaporization method. T-Hg concentrations for the PBs with diameters of 0.02, 0.2, and 2 µm were 10.6 ± 0.4, 1.8 ± 0.1, and 1.3 ± 0.1 ng/mg-PBs, respectively, when immersed in 2 mL of MeHg-added seawater (100 µg/L as Hg). Thus, the adsorption efficiency of MeHg onto PBs was higher in the presence of smaller diameter PBs. Next, 1 mg of PBs immersed in 2 mL of seawater containing 100 µg/L of MeHg for 24 h was added to an oyster tank containing 1 L of seawater. The T-Hg concentration of the oysters was measured after 6 h of exposure. No significant difference was found in the T-Hg concentration of oysters in the presence of PBs (0.30 ± 0.01 to 0.37 ± 0.05 ng/mg as dry weight) with MeHg and in the absence of PBs (0.36 ± 0.03 ng/mg as dry weight). Our results suggest that the presence of PBs in seawater has little effect on MeHg uptake by oysters.

Shoot has important roles in strigolactone production of rice roots under sulfur deficiency.

Strigolactones (SLs) are a class of plant hormones that control plant architecture. SL levels in roots are determined by the nutrient conditions in the rhizosphere, especially the levels of nitrogen (N) and phosphorus (P). Our previous research showed that SL production is induced in response to deficiency of sulfur (S) as well as of N and P, and inhibits shoot branching, accelerates leaf senescence, and regulates lamina joint angle in rice. Here we show biomass, total S contents, and SL levels in rice under S-sufficient and S-deficient conditions using a split-root system. When one part of the root system was cultured in S-sufficient medium and the other in S-deficient medium (+S/-S), shoot fresh weight was unaffected relative to the +S/+S condition. The shoot weight significantly decreased in -S/-S condition. In contrast, there was no significant difference in root fresh weight between +S and -S conditions. In +S/-S condition, SL levels were systemically reduced in both parts, the shoot S content increased, but the root S content in S-deficient medium was unaffected relative to the -S/-S condition. These results suggest that shoots, not roots, recognize S deficiency, which induces SL production in roots.

Silver nanocolloid affects hindbrain vascular formation during medaka embryogenesis.

Angiogenesis is essential for the normal development of an embryo. Silver nanocolloid (SNC) is known to induce vascular malformation in the medaka embryo. We focused on the development of the central arteries (CtAs) in the hindbrain of Japanese medaka. The CtAs and the basilar artery from which they branch are essential for transporting the blood and nutrients necessary to support the hindbrain parenchyma and the development of the pons and cerebellum from the hindbrain. We exposed medaka embryos at developmental stage 21 (6 somite stage), to 0, 0.5, 5, or 10 mg/L SNC and evaluated hatching rate, number of thrombi per embryo, head size (length and width), body length, and angiogenesis. Although all SNC-exposed embryos hatched, their head size and body length were small in comparison to controls; in addition, the number of thrombi in the head increased and head size and body length decreased as the SNC concentration increased. To evaluate vasculogenic abnormalities, we performed whole-mount in situ hybridization using a vascular marker (eg, fl7) and visualized the CtAs in medaka embryos. In control embryos, CtAs started to sprout at stage 32 (somite completion stage) and their extension was complete by stage 35 (pectoral fin blood circulation stage). In contrast, CtAs failed to sprout in SNC-exposed embryos, and thrombi were present. Furthermore, qRT-PCR analysis showed that SNC significantly suppressed the egfl7 expression level at stage 35. Together, our findings suggest that SNC induced decreased developments of head and body in medaka embryos due to insufficient angiogenesis and hindbrain vascular formation.

γ-H2AX foci as indication for the DNA damage in erythrocytes of medaka (Oryzias latipes) intoxicated with 4-nonylphenol.

The present study aimed to investigate the genotoxicity in erythrocytes induced after exposure of medaka (Oryzias latipes) to 4-nonylphenol (4-NP). Adult female medaka fish were exposed to 4-NP at three sublethal concentrations for 15 days to compare their sensitivity with that of catfish as an aquatic model. Comet assay and γ-H2AX were used as biomarkers to detect DNA damage in erythrocytes. Exposure to 4-NP resulted in an increase in the tail moment in a dose-dependent manner. The highest level of DNA damage was recorded after exposure to 100 µg/l 4-NP. The number of foci was increased after exposure to 4-NP, indicating damage to DNA. The present results confirmed the high level of morphological alterations and apoptosis of erythrocytes detected in the first part of this study. 4-NP induced genotoxic effects in medaka, which were found to be more sensitive than catfish after exposure to 4-nonylphenol. Graphical abstract.

Temperature-dependent toxicity of acetaminophen in Japanese medaka larvae.

Because of its analgesic properties, acetaminophen (AAP) is widely used to relieve headache. AAP is generally considered safe for humans, but its effects on aquatic organisms are not well known. Here, we have hypothesis that effects of AAP on aquatic organisms would be environmental temperature dependent, because their physiological function depend on the temperature. To test this hypothesis, we used medaka (Oryzias latipes) as a model, because they can live at a wide range of temperatures (0-40 °C). We exposed medaka larvae to 0 (control), 50, or 150 mg/L of AAP at 15, 25 (optimal temperature), or 30 °C for 4 days. Egg yolk absorption was accelerated with raising temperature at any AAP dose. AAP exposure did not have biologically significant effects on survival ratio and body length of larvae at any tested temperature or dose, but heart rate decreased as the dose of AAP and environmental temperature increased. In addition, as the temperature increased, amount of ATP in individual larvae increased in control group, but decreased in AAP exposed group. Subsequently, exposure to 150 mg/L of AAP at 30 °C decreased the number of red blood cells in the gills; we used 150 mg/L of AAP in subsequent hematological and histological analyses. Hematological analysis showed that rising temperature increased the proportion of morphologically abnormal red blood cells in AAP-exposed larvae, suggesting that AAP induced anemia-like signs in larvae. Histological observation of the kidney, which is a hematopoietic organ in fish, revealed no abnormalities. However, in the liver, which is responsible for drug metabolism, the proportion of vacuoles increased with increasing temperature. Although the exposure concentration we tested was higher than environmentally relevant concentrations, our data indicated that rising temperature enhances the toxicity of AAP to medaka larvae, suggesting an ecological risk of AAP due to global warming.

Protective effect of p53 knockout on 4-nonylphenol-induced nephrotoxicity in medaka (Oryzias latipes).

In the past few decades, environmental pollutants have become common because of misused nonionic surfactants and detergents. Nonylphenol ethoxylates (NPs) are one of the most important contaminants of water. Therefore, the present study aimed to investigate the protective blocking effect of apoptosis (deficient P53 gene) on 4-nonylphenol (4-NP)-induced nephrotoxicity of medaka (Oryzias latipes). We divided 36 fish into six groups: two different control groups of wild type (Wt; Hd-rR) control and p53 (-/-) control, and four different treated with 4-nonylphenol (50 µg/L and 100 µg/L) for 15 days. Histology, immunochemistry, and TUNEL assays confirmed that 4-NP causes nephrotoxicity. Our results showed that 4-NP administration significantly disturbed the kidney structure and function and 4-NP-treated fish showed dilated glomerular vessels, had less glomerular cellular content, decreased expression of glomerular proteins, and an increased level of apoptosis compared with a Wt control group (P < 0.05). As p53 is an apoptotic inducer, some protection in p53-deficient medaka was found as nephrotoxic effects of 4-NP were minimized significantly. Our study demonstrated for the first time to our knowledge that 4-NP induces apoptosis, causing nephrotoxicity in medaka. We found that blocking apoptosis blocking was able to protect the kidney from the toxic effects of 4-NP.

Extracellular vesicles from human bone marrow mesenchymal stem cells repair organ damage caused by cadmium poisoning in a medaka model.

Treatment modalities for kidney disease caused by long-term exposure to heavy metals, such as cadmium (Cd), are limited. Often, chronic, long-term environmental exposure to heavy metal is not recognized in the early stages; therefore, chelation therapy is not an effective option. Extracellular vesicles (EVs) derived from stem cells have been demonstrated to reduce disease pathology in both acute and chronic kidney disease models. To test the ability of EVs derived from human bone marrow mesenchymal stem cells (hBM-MSCs) to treat Cd damage, we generated a Cd-exposed medaka model. This model develops heavy metal-induced cell damage in various organs and tissues, and shows decreased overall survival. Intravenous injection of highly purified EVs from hBM-MSCs repaired the damage to apical and basolateral membranes and mitochondria of kidney proximal tubules, glomerular podocytes, bone deformation, and improved survival. Our system also serves as a model with which to study age- and sex-dependent cell injuries of organs caused by various agents and diseases. The beneficial effects of EVs on the tissue repair process, as shown in our novel Cd-exposed medaka model, may open new broad avenues for interventional strategies.

Cytotoxic and genotoxic effects of arsenic on erythrocytes of Oryzias latipes: Bioremediation using Spirulina platensis.

BACKGROUND: Exposure to the environmental pollutants poses a serious threat to aquatic organism. The arsenic exposure in fish increases the risk of developing serious alterations from embryo to adult. OBJECTIVES: The present investigation was done to study the toxic effects of heavy metal arsenic [As(III)] on medaka (Oryzias latipes). Morphological alterations, apoptosis, nuclear abnormalities, and genotoxic biomarkers in erythrocytes were used to determine the stress caused by arsenic (As) exposure. METHODS: Medaka was exposed to As for 15 days at two toxic sublethal concentrations (7 ppm and 10 ppm) in combination with Spirulina platensis (SP) treatment as antioxidant algae at 200 mg/L. RESULTS: Results were consistent with a previous study results on tilapia. Exposure of medaka to As resulted in a dose-dependent increase in most the biomarkers used in the current study. Fish exposed to10 ppm As showed highest level of DNA damage. For the first time to our knowledge, using SP to counter the As toxicity in medaka, DNA damage restored to control levels. CONCLUSION: Accordingly, those results suggests that SP can protect medaka in aquaculture against As-induced damage by its ability as reactive oxygen species (ROS) reducer, antioxidant role, and DNA damage scavenger.

Introducing a new standardized nanomaterial environmental toxicity screening testing procedure, ISO/TS 20787: aquatic toxicity assessment of manufactured nanomaterials in saltwater Lakes using Artemia sp. nauplii.

This paper introduces a new standardized testing procedure for nanomaterial environmental toxicity (International Organization for Standardization/Technical Specification (ISO/TS) 20787): 'aquatic toxicity assessment of manufactured nanomaterials in saltwater lakes using Artemia sp. Nauplii' intended to generate more reliable and repeatable aquatic toxicity data testing manufactured nanomaterials, using Artemia sp., to evaluate their possible ecotoxicity in saltwater lake ecosystems. The principles behind testing with Artemia sp. are reviewed and the paper gives an overview of research published between 2009 and 2018 in which manufactured nanomaterials were tested using Artemia sp.

Sensitivity of medaka (Oryzias latipes) to 4-nonylphenol subacute exposure; erythrocyte alterations and apoptosis.

The present study was undertaken to assess the effects of the endocrine-disrupting compound; 4-nonylphenol (4-NP) in medaka (Oryzias latipes). The frequencies of erythrocyte alterations, apoptosis, and micronuclei were used as biological indicators of damage. Medaka were exposed 15 days to 4-NP at three sublethal concentrations (50, 80, and 100 µg/l 4-NP) and results compared with those of a previous study using catfish as an animal model. Exposure of medaka resulted in a dose-dependent increase in the frequency of erythrocyte alterations, apoptosis and micronucleus (MN). Many morphological alterations and nuclear abnormalities were observed, including acanthocytes, lobed nucleus, eccentric nucleus, fragmented nucleus, blebbed nucleus, binuclei, deformed nucleus, notched nucleus, hemolysed cells, crenated cells, teardrop-like cells, and schistocytes. Mortality was recorded after treatment with 80 and 100 µg/l 4-NP, indicating that medaka are more sensitive than catfish to 4-NP exposure. We concluded that, 4-NP causes several malformations in the shape and number of erythrocytes in medaka, indicating its genotoxicity.

Effects of silver nanocolloids on plant complex type N-glycans in Oryza sativa roots.

Silver nanomaterials have been mainly developed as antibacterial healthcare products worldwide, because of their antibacterial activity. However, there is little data regarding the potential risks and effects of large amounts of silver nanomaterials on plants. In contrast, N-glycans play important roles in various biological phenomena, and their structures and expressions are sensitive to ambient environmental changes. Therefore, to assesse the effects of silver nanomaterials, we focused on the correlation between N-glycans and the effects of silver nanomaterials in plants and analyzed N-glycan structures in Oryza sativa seedlings exposed to silver nanocolloids (SNCs). The phenotype analysis showed that the shoot was not affected by any SNC concentrations, whereas the high SNC exposed root was seriously damaged. Therefore, we performed comparative N-glycan analysis of roots. As a result, five of total N-glycans were significantly increased in SNC exposed roots, of which one was a free-N-glycan with one beta-N-acetylglucosamine residue at the reducing end. Our results suggest that the transition of plant complex type N-glycans, including free-N-glycans, was caused by abnormalities in O. sativa development, and free-N-glycan itself has an important role in plant development. This study originally adapted glycome transition analysis to environmental toxicology and proposed a new category called "Environmental glycobiology".

Comparative toxicities of silver nitrate, silver nanocolloids, and silver chloro-complexes to Japanese medaka embryos, and later effects on population growth rate.

Fish embryo toxicology is important because embryos are more susceptible than adults to toxicants. In addition, the aquatic toxicity of chemicals depends on water quality. We examined the toxicities to medaka embryos of three types of silver-AgNO3, silver nanocolloids (SNCs), and silver ions from silver nanoparticle plates (SNPPs)-under three pH conditions (4.0, 7.0, and 9.0) in embryo-rearing medium (ERM) or ultrapure water. Furthermore, we tested the later-life-stage effects of SNCs on medaka and their population growth. "Later-life-stage effects" were defined here as delayed toxic effects that occurred during the adult stage of organisms that had been exposed to toxicant during their early life stage only. AgNO3, SNCs, and silver ions were less toxic in ERM than in ultrapure water. Release of silver ions from the SNPPs was pH dependent: in ERM, silver toxicity was decreased owing to the formation of silver chloro-complexes. SNC toxicity was higher at pH 4.0 than at 7.0 or 9.0. AgNO3 was more toxic than SNCs. To observe later-life effects of SNCs, larvae hatched from embryos exposed to 0.01 mg/L SNCs in ultrapure water were incubated to maturity under clean conditions. The mature medaka were then allowed to reproduce for 21 days. Calculations using survival ratios and reproduction data indicated that the intrinsic population growth rate decreased after exposure of embryos to SNC. SNC exposure reduced the extinction time as a function of the medaka population-carrying capacity.