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Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues

Silver nanoparticles (AgNPs) have been extensively used in commercial products, including textiles, cosmetics, and health care items, due to their strong ...

National Sun Yat-sen University

Baseline metal concentrations in sediments and fish, and the determination of bioindicators in the subtropical Chi-ku Lagoon, S W Taiwan.

Total Hg (sigmaHg), organic-Hg (O-Hg) and Se bioaccumulations in small cetaceans distributed in Taiwanese waters of the Taiwan Strait and the southwestern Pacific have been investigated for the first time. The results could represent the baseline metal concentrations of marine mammals in the southwestern Pacific, where volcanic activities are possibly the major source of mercury to the environments. Muscle samples of four species of small cetaceans were collected from animals accidentally caught by tuna-longline fisheries from 1994 through 1995. In total, 53 pantropical spotted dolphins, Stenella attenuata, nine spinner dolphins, S. longirostris, five bottlenose dolphins, Turiops truncatus and four Risso's dolphins, Grampus giseus were analyzed. In addition, two stranded pantropical spotted dolphins were investigated. Cold vapour AAS and ICP-MS were used in the analysis of Hg and Se, respectively. Significant species difference was found in the four species of small cetaceans. Among them, the pantropical spotted dolphin showed the highest mean concentration (mg/kg wet wt.) of both sigmaHg (3.64 +/- 2.19) and O-Hg (2.79 +/- 1.23), whereas the Risso's dolphin had the highest mean concentrations of Se (1.77 +/- 1.29). There was no significant sex difference with respect to metal bioaccumulation in the samples of S. attenuata. Significant correlations between body length (BL) and sigmaHg, as well as O-Hg concentrations were observed in pantropical spotted (Sa) and spinner dolphins (Sl). The linear relationships were Sa: sigmaHg = -8.290 + 0.066BL, r = 0.421; Sl: sigmaHg = -2.735 + 0.025BL, r = 0.875; Sa: O-Hg = -3.723 + 0.036BL, r = 0.408; and Sl: O-Hg = -3.017 + 0.025BL, r = 0.870. However, a demethylation phenomenon that decreasing the percentage of O-Hg coupled with increasing levels of Se was observed when the sigmaHg concentrations in the muscle tissues of dolphins reached 4 mg/kg wet wt.

Mercury, organic-mercury and selenium in small cetaceans in Taiwanese waters.

Chemical investigation of the gorgonian coral Junceella juncea collected in Taiwan has resulted in the isolation of the two novel briarane-type diterpenoid compounds, juncenolides F (1) and G (2). The structures were determined on the basis of spectral studies, especially 1D and 2D NMR.

Juncenolides F and G, two new briarane diterpenoids from Taiwanese gorgonian Junceella juncea.

Gender and size effects of metal bioaccumulation on the rock crab, Thalamita crenata, in Dapeng Bay, southwestern Taiwan.

Muscles of 115 North Pacific albacore (ALB, Thunnus alalunga) and 75 Pacific bigeye tuna (BET, Thunnus obesus), collected from 2001 to 2006, were analyzed. No ALB, but 13 large BET had organic mercury (OH g) concentrations exceeding 1 μg g(-1) wet weight. For both ALB and BET, total mercury (THg) and OH g concentrations were significantly and positively correlated with fork length (FL) and body weight. The muscle Hg bioaccumulation rates of BET were higher than those of ALB, particularly in the adult fish. Moreover, the lines had crossover points among the two species that imply the young BET (FL<110 cm) contains lower muscle Hg concentrations than ALB of the same size. The suggested weekly dietary intake of ALB and small-BET meats is 340 g, and of BET meat it is 150 g for a 60-kg person based on the provisional tolerable weekly intake (PTWI) of methylmercury set by the WHO.

Total and organic mercury concentrations in the muscles of Pacific albacore (Thunnus alalunga) and bigeye tuna (Thunnus obesus).

We describe the complete mitochondrial genome sequence of Rasbora trilineata, which is a small cyprinid popular in aquarium trade. The circle genome (16,747 bp) has the typical vertebrate mitochondrial gene arrangement, including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a non-coding control region. The overall base composition of R. trilineata is 25.35% for T, 26.43% for C, 33.57% for A, and 14.65% for G, with a slight AT bias of 58.92%.

Complete mitochondrial genome of Rasbora trilineata (Cypriniformes, Cyprinidae).

Indo-Pacific humpback dolphins (Sousa chinensis) have attracted considerable attention due to their critically endangered status and related conservation issues, but their trophic relationships and ecological significance in coastal ecosystems are poorly understood. For instance, this species is noticeably more abundant in the Xin-Huwei River Estuary (Ex) of Western Taiwan than in the nearby Zhuoshui River Estuary (Ez), though it is unclear why the distribution shows such partitioning. To explore this topic, we conducted field surveys seasonally for two years from 2012 to 2013 and constructed Ecopath models of Ex, Ez, and an offshore site (Dm) to compare energy flow within the food webs. Model comparisons showed that the availability of food resources was the main factor influencing the biomass of Indo-Pacific humpback dolphins. Specifically, its more frequent occurrence in Ex can be attributed to greater phytoplankton production and greater biomasses of macroinvertebrates and prey fish than in the other two areas. An increase in fishing activity might decrease the food availability and, consequently, the biomass of the dolphins. Although the decline in the dolphin population would increase the biomass of some prey fish species, local fishermen might not necessarily benefit from the decline due to the concurrent decrease of highly valued crabs and shrimp. Collectively, our work suggests that the Indo-Pacific humpback dolphin is a keystone species in tropical coastal waters of Taiwan, and thereby exhibit a disproportional large ecological impact given their relatively low abundance.

The Trophic Significance of the Indo-Pacific Humpback Dolphin, Sousa chinensis, in Western Taiwan.

Muscle, lung, kidney and liver tissues of 45 bycatch and stranded cetaceans, including 14 Grampus griseus (Gg), 7 Kogia simus (Ks), 10 Lagenodelphis hosei (Lh), and 14 Stenella attenuata (Sa), were collected in the waters off Taiwan from 1994 to 1995, and from 2001 to 2012. Baseline concentrations (in μgg dry weight) of the cetaceans were lung (<0.05)=muscle (<0.05)<kidney (0.08±0.04)<liver (0.43±0.28) for Ag, and muscle (0.03±0.03)=lung (0.22±0.19)<liver (3.82±3.50)<kidney (16.22±18.81) for Cd. Unhealthy and critically dangerous Ag and Cd tissue concentrations in the toothed cetaceans are suggested. Marked high concentrations of Ag and Cd found in Gg and Lh are highly related to their squid-eating and deep diving habits. The highest ever recorded concentrations of liver-Ag and kidney-Cd were found in two Lh. These Taiwanese cetaceans indicate marked Ag and Cd pollution in the recent two decades in the western Pacific Ocean.

Tissue concentrations of four Taiwanese toothed cetaceans indicating the silver and cadmium pollution in the western Pacific Ocean.

Silver, such as silver nanoparticles (AgNPs), has been widely used in commercial products and may be released into the environment. The interaction between Ag deposition and biological systems is raising serious concerns because of one health consideration. Cetaceans, as the top predators of the oceans, may be exposed to Ag/Ag compounds and suffer negative health impacts from the deposition of these compounds in their bodies. In the present study, we utilized autometallography (AMG) to localize the Ag in the liver and kidney tissues of cetaceans and developed a model called the cetacean histological Ag assay (CHAA) to estimate the Ag concentrations in the liver and kidney tissues of cetaceans. Our results revealed that Ag was mainly located in hepatocytes, Kupffer cells and the epithelial cells of some proximal renal tubules. The tissue pattern of Ag/Ag compounds deposition in cetaceans was different from those in previous studies conducted on laboratory rats. This difference may suggest that cetaceans have a different metabolic profile of Ag, so a presumptive metabolic pathway of Ag in cetaceans is advanced. Furthermore, our results suggest that the Ag contamination in cetaceans living in the North-western Pacific Ocean is more severe than that in cetaceans living in other marine regions of the world. The level of Ag deposition in cetaceans living in the former area may have caused negative impacts on their health condition. Further investigations are warranted to study the systemic Ag distribution, the cause of death/stranding, and the infectious diseases in stranded cetaceans with different Ag concentrations for comprehensively evaluating the negative health effects caused by Ag in cetaceans.

Investigation of silver (Ag) deposition in tissues from stranded cetaceans by autometallography (AMG).

Silver nanoparticles (AgNPs) have been extensively used and are considered as an emerging contaminant in the ocean. The environmental contamination of AgNPs is expected to increase greatly over time, and cetaceans, as the top ocean predators, will suffer the negative impacts of AgNPs. In the present study, we investigate the immunotoxicity of AgNPs on the leukocytes of cetaceans using several methods, including cytomorphology, cytotoxicity, and functional activity assays. The results reveal that 20 nm Citrate-AgNPs (C-AgNP) induce different cytomorphological alterations and intracellular distributions in cetacean polymorphonuclear cells (cPMNs) and peripheral blood mononuclear cells (cPBMCs). At high concentrations of C-AgNP (10 and 50 μg/ml), the time- and dose-dependent cytotoxicity in cPMNs and cPBMCs involving apoptosis is demonstrated. C-AgNP at sub-lethal doses (0.1 and 1 μg/ml) negatively affect the functional activities of cPMNs (phagocytosis and respiratory burst) and cPBMCs (proliferative activity). The current study presents the first evidence of the cytotoxicity and immunotoxicity of AgNPs on the leukocytes of cetaceans and improves our understanding of environmental safety concerning AgNPs. The dose-response data of AgNPs on the leukocytes of cetaceans are invaluable for evaluating the adverse health effects in cetaceans and for proposing a conservation plan for marine mammals.

Immunotoxicity of Silver Nanoparticles (AgNPs) on the Leukocytes of Common Bottlenose Dolphins (Tursiops truncatus).

Silver nanoparticles (AgNPs) have been widely used in many commercial products due to their excellent antibacterial ability. The AgNPs are released into the environment, gradually accumulate in the ocean, and may affect animals at high trophic levels, such as cetaceans and humans, via the food chain. Hence, the negative health impacts caused by AgNPs in cetaceans are of concern. Cytokines play a major role in the modulation of immune system and can be classified into two types: Th1 and Th2. Th1/Th2 balance can be evaluated by the ratios of their polarizing cytokines (i.e., interferon [IFN]-γ/Interleukin [IL]-4), and animals with imbalanced Th1/Th2 response may become more susceptible to certain kinds of infection. Therefore, the present study evaluated the  cytokine responses of cetacean peripheral blood mononuclear cells (cPBMCs) to 20 nm citrate-AgNPs (C-AgNP) by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR).

Meng-Hsien Chen

Department of Oceanography and Asia-Pacific Ocean Research Center

National Sun Yat-sen University

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues

Meng-Hsien Chen

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Oceanography and Asia-Pacific Ocean Research Center

National Sun Yat-sen University

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues

Department of Oceanography and Asia-Pacific Ocean Research Center