A 2 °C difference affecting the spatiotemporal distribution of small demersal fish assemblages in shallow tropical and subtropical waters of Western Taiwan.

Two OR3 research vessel cruises were conducted at seven nearshore sites from north to south in the western coastal waters off Taiwan in May (late spring) and November (late autumn) 2019 in order to gain insights into the 2 °C difference in the tropical-subtropical fish fauna. Totally, 37 families, 72 genera, and 99 taxas were recorded for the 1809 fishes. Three fish assemblages, the North, South, and Fall-Zhuoshui River (ZRf) groups, were identified as eurythermal, stenothermal, and croaker communities. Their dominants, in rank order, were Tarphops oligolepis, Liachirus melanospilosa, Ostorhinchus fasciatus, and Trachinocephalus myops for the Northern eurythermal group, Arnoglossus tenuis, Eubleekeria splendens, and Ostorhinchus pleuron for the Southern stenothermal group, and Johnius taiwanensis, Chrysochir aureus, and Pennahia macrocephalus for the croaker-ZRf group. Their distribution was markedly correlated with the bottom water temperature, seafloor grain size, and concentration of suspended solids. The influence of a 2 °C difference on the tropical and subtropical demersal fish could indicate how rising temperatures due to climate change are shaping fish communities. The flatfish, Liachirus melanospilos, was distributed northward by 0.5oN and is suggested to be considered as an ecological indicator of the tropicalization of subtropical marine ecosystems in the future.

Johnius sasakii, a new species of croaker (Perciformes: Sciaenidae) with a key to Johnius from East Malaysia, Borneo.

A new sciaenid fish, Johnius sasakii sp. nov. from the East Malaysian coastal waters of Sabah and Sarawak, Borneo is described herein. Johnius sasakii sp. nov. can be separated from the close congeners, Johnius heterolepis and Johnius carouna by having less gill rakers on the lower limb of the first gill arch (mode number 9 vs. 10 for J. heterolepis; 11 for J. carouna). It can be distinguished from J. heterolepis with the higher modal number of gill rakers on the upper limb of first gill arch (5 vs. 4), shorter anal-fin base length (9% vs. 10% of SL) and first anal-fin ray length (1012% vs. 1213% of SL) respectively. Furthermore, J. sasakii sp. nov. can be further differentiated from J. carouna by its shorter second anal-fin spine length 710% SL (usually 810%) vs. 1114% SL (usually 1112%) and slightly deeper body depth 2529% SL (usually 2628%) vs. 2328% SL (usually 2426%). Meanwhile, J. sasakii sp. nov. can be easily differentiated from J. macrorhynus by a present of molariform teeth on inner rows of jaws and stumpy gill rakers. Johnius (Johnius) sasakii sp. nov. can be clearly distinguished from species of the subgenus J. (Johnieops) by the lack of enlarged outer row teeth on upper jaw. A Kimura 2-parameter genetic distance comparison of 450 bp sequences of mitochondrial 16S rRNA and 488 bp nuclear gene S7 showed that J. sasakii sp. nov. are highly differentiated from J. heterolepis (16S: 10.4%, S7: 5.8%), J. carouna (16S:19.3%, S7:8.4%); and J. macrorhynus (16S: 16.7%, S7: 8.1%) respectively. The study highlights that the genetic approach from mtDNA and nDNA can contribute to the confirmation of taxonomic status of sympatric species in genus Johnius.

Tissue concentrations of Fe, Zn, Cu and Mn of four Taiwanese toothed cetaceans.

Concentrations of Fe, Mn, Zn, and Cu in the muscle, lung, liver, and kidney tissues of 49 cetaceans, including 11 Kogia sima (Ks), 10 Lagenodelphis hosei (Lh), 14 Grampus griseus (Gg), and 14 Stenella attenuata (Sa) from 1994 to 2012 in Taiwan were measured. Ks exhibited the highest Fe, and the lowest Zn, Cu and Mn tissue concentrations. The Kogiid and Delphinid groups were significantly categorized by nMDS analysis. Fe muscle concentrations were found to be positively correlated with Ks, Lh, and Gg but not Sa body length. The different levels of Zn-, Cu-, and Mn- tissue concentrations of the two families were due to their different SOD systems. Their calves contained higher Cu liver concentrations. The four essential elements in cetaceans regulate homeostasis to meet their eco-physiological demand. The baseline levels for these four elements in these four tissues in the Delphinid group are defined.

Johnius taiwanensis, a new species of Sciaenidae from the Taiwan Strait, with a key to Johnius species from Chinese waters.

A new sciaenid fish, Johnius taiwanensis, is described from the southeast coast of mainland China from Zhejiang to Guangdong, Hong Kong, and west coast of Taiwan. Johnius taiwanensis sp. nov. can be distinguished from other Johnius species by having a grayish dorsal half of body divided by a clear line from a whitish ventral half, and a black spot at the dorsal half of pectoral-fin axil, appearing as a distinct dot at the most dorsal point of the pectoral-fin base. First dorsal fin black tipped, other fins pale to dusky but never darkly pigmented. The species lacks distinctly enlarged teeth on upper and lower jaws. Body scales ctenoid, moderately large, with five or six rows between first dorsal-fin origin and lateral line. It is one of the most abundant sciaenids found in the shallow coastal waters (20 m) of southeast mainland China and the west coast of Taiwan. It has often been misidentified as J. macrorhynus in the region. Phylogenetic analysis from all 27 sciaenid species found in Chinese waters based on the complete COI and 16S rRNA gene sequences confirmed that the genus Johnius is monophyletic and J. taiwanensis is placed as a sister species of J. trewavasae. Acoustic analysis has shown that J. taiwanensis produces a unique sound among fishes in Taiwan coastal waters.

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 antimicrobial effects. They also may be released into the environment and accumulate in the ocean. Therefore, AgNPs are the major source of Ag contamination, and public awareness of the environmental toxicity of Ag is increasing. Previous studies have demonstrated the bioaccumulation (in producers) and magnification (in consumers/predators) of Ag. Cetaceans, as the apex predators of ocean, may have been negatively affected by the Ag/Ag compounds. Although the concentrations of Ag/Ag compounds in cetacean tissues can be measured by inductively coupled plasma mass spectroscopy (ICP-MS), the use of ICP-MS is limited by its high capital cost and the requirement for tissue storage/preparation. Therefore, an autometallography (AMG) method with an image quantitative analysis by using formalin-fixed, paraffin-embedded (FFPE) tissue may be an adjuvant method to localize Ag distribution at the suborgan level and estimate the Ag concentration in cetacean tissues. The AMG positive signals are mainly brown to black granules of various sizes in the cytoplasm of proximal renal tubular epithelium, hepatocytes, and Kupffer cells. Occasionally, some amorphous golden yellow to brown AMG positive signals are noted in the lumen and basement membrane of some proximal renal tubules. The assay for estimating the Ag concentration is named the Cetacean Histological Ag Assay (CHAA), which is a regression model established by the data from image quantitative analysis of the AMG method and ICP-MS. The use of AMG with CHAA to localize and semi-quantify heavy metals provides a convenient methodology for spatio-temporal and cross-species studies.

Th2 cytokine bias induced by silver nanoparticles in peripheral blood mononuclear cells of common bottlenose dolphins (Tursiops truncatus).

BACKGROUND: 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 in vitro cytokine responses of cetacean peripheral blood mononuclear cells (cPBMCs) to 20 nm citrate-AgNPs (C-AgNP20) by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). METHODS: Blood samples were collected from six captive common bottlenose dolphins (Tursiops truncatus). The cPBMCs were isolated and utilized for evaluating the in vitro cytokine responses. The cytokines evaluated included IL-2, IL-4, IL-10, IL-12, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α. The geometric means of two housekeeping genes (HKGs), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and ß2-microglobulin (B2M), of each sample were determined and used to normalize the mRNA expression levels of target genes. RESULTS: The ratio of late apoptotic/necrotic cells of cPBMCs significantly increased with or without concanavalin A (ConA) stimulation after 24 h of 10 µg/ml C-AgNP20 treatment. At 4 h of culture, the mRNA expression level of IL-10 was significantly decreased with 1 µg/ml C-AgNP20 treatment. At 24 h of culture with 1 µg/ml C-AgNP20, the mRNA expression levels of all cytokines were significantly decreased, with the exceptions of IL-4 and IL-10. The IFN-γ/IL-4 ratio was significantly decreased at 24 h of culture with 1 µg/ml C-AgNP20 treatment, and the IL-12/IL-4 ratio was significantly decreased at 4 or 24 h of culture with 0.1 or 1 µg/ml C-AgNP20 treatment, respectively. Furthermore, the mRNA expression level of TNF-α was significantly decreased by 1 µg/ml C-AgNP20 after 24 h of culture. DISCUSSION: The present study demonstrated that the sublethal dose of C-AgNP20 (≤1 µg/ml) had an inhibitory effect on the cytokine mRNA expression levels of cPBMCs with the evidence of Th2 cytokine bias and significantly decreased the mRNA expression level of TNF-α. Th2 cytokine bias is associated with enhanced immunity against parasites but decreased immunity to intracellular microorganisms. TNF-α is a contributing factor for the inflammatory response against the infection of intracellular pathogens. In summary, our data indicate that C-AgNP20 suppresses the cellular immune response and thereby increases the susceptibility of cetaceans to infection by intracellular microorganisms.

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

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-AgNP20) induce different cytomorphological alterations and intracellular distributions in cetacean polymorphonuclear cells (cPMNs) and peripheral blood mononuclear cells (cPBMCs). At high concentrations of C-AgNP20 (10 and 50 µg/ml), the time- and dose-dependent cytotoxicity in cPMNs and cPBMCs involving apoptosis is demonstrated. C-AgNP20 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.

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

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.

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

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-1 dry weight) of the cetaceans were lung (<0.05)=muscle (<0.05)

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

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.

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

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%.

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

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.

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

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.

Application of acetylacetone chelation solid-phase extraction to GFAAS measurements of trace amounts of beryllium in marine organisms.

Trace amounts of Be (0.046-2.59 ng) in a dried marine organism sample (10 mg) could be accurately determined by GFAAS after treating with microwave digestion (HNO3/H2O2) at 85 degrees C for 10 min and using acetylacetone as a chelating agent in the presence of an acetate buffer (pH 6.0). The method detection limit (MDL, 3sigma) for Be was found to be 4.6 ng g(-1); the calibration graph was linear up to 259 ng g(-1). Good recoveries (98.5-105.0%) were obtained for eight marine organism samples (including five fish, one lobster, one oyster, and one algae) with a relative standard deviation (RSD, n = 3) < 3.0%. The proposed method could be applied measurements of Be in various marine organisms.

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

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.