Xiwu Yan

Seawater acidification affects the physiological energetics and spawning capacity of the Manila clam Ruditapes philippinarum during gonadal maturation.

Ocean acidification is predicted to have widespread implications for marine bivalve mollusks. While our understanding of its impact on their physiological and behavioral responses is increasing, little is known about their reproductive responses under future scenarios of anthropogenic climate change. In this study, we examined the physiological energetics of the Manila clam Ruditapes philippinarum exposed to CO2-induced seawater acidification during gonadal maturation. Three recirculating systems filled with 600 L of seawater were manipulated to three pH levels (8.0, 7.7, and 7.4) corresponding to control and projected pH levels for 2100 and 2300. In each system, temperature was gradually increased ca. 0.3°C per day from 10 to 20°C for 30days and maintained at 20°C for the following 40days. Irrespective of seawater pH levels, clearance rate (CR), respiration rate (RR), ammonia excretion rate (ER), and scope for growth (SFG) increased after a 30-day stepwise warming protocol. When seawater pH was reduced, CR, ratio of oxygen to nitrogen, and SFG significantly decreased concurrently, whereas ammonia ER increased. RR was virtually unaffected under acidified conditions. Neither temperature nor acidification showed a significant effect on food absorption efficiency. Our findings indicate that energy is allocated away from reproduction under reduced seawater pH, potentially resulting in an impaired or suppressed reproductive function. This interpretation is based on the fact that spawning was induced in only 56% of the clams grown at pH 7.4. Seawater acidification can therefore potentially impair the physiological energetics and spawning capacity of R. philippinarum.

Heavy Metal Concentrations in Surface Sediments and Manila Clams (Ruditapes philippinarum) from the Dalian Coast, China after the Dalian Port Oil Spill

We conducted an investigation of heavy metal concentrations in Manila clams (Ruditapes philippinarum) and surface sediments after the Dalian Port oil spill. Samples were collected from three mariculture zones (Jinshitan, Dalijia, and Pikou) along the Dalian coast. Heavy metal concentrations in R. philippinarum were consistent and ranked in decreasing order of Zn > Cu > As > Cr > Pb > Cd > Hg, while concentrations in surface sediments were ranked as Zn > Cr > Cu > Pb > As > Cd > Hg, respectively. Bioaccumulation of Zn, Cd, and Hg had obviously occurred in R. philippinarum. Statistically significant correlations (p < 0.05) between concentrations of Pb, Cd, and Hg in R. philippinarum and in surface sediments were observed. Except for Cr and As, heavy metal concentrations in R. philippinarum were well within the legal limits for human consumption.

Bioaccumulation of Trace Elements in Ruditapes philippinarum from China: Public Health Risk Assessment Implications

The Manila clam Ruditapes philippinarum is one of the most important commercial bivalve species consumed in China. Evaluated metal burden in bivalve molluscs can pose potential risks to public health as a result of their frequent consumption. In this study, concentrations of 10 trace elements (Cu, Zn, Mn, Se, Ni, Cd, Cr, Pb, Hg and As) were determined in samples of the bivalve Ruditapes philippinarum, collected from nine mariculture zones along the coast of China between November and December in 2010, in order to evaluate the status of elemental metal pollution in these areas. Also, a public health risk assessment was untaken to assess the potential risks associated with the consumption of clams. The ranges of concentrations found for Cu, Zn, Mn, Se, Ni, Cd, Cr, Pb, Hg and As in R. philippinarum were 12.1–38.0, 49.5–168.3, 42.0–68.0, 4.19–8.71, 4.76–14.32, 0.41–1.11, 0.94–4.74, 0.32–2.59, 0.03–0.23 and 0.46–11.95 mg·kg−1 dry weight, respectively. Clear spatial variations were found for Cu, Zn, Cr, Pb, Hg and As, whereas Mn, Se, Ni, and Cd did not show significant spatial variation. Hotspots of trace element contamination in R. philippinarum can be found along the coast of China, from the north to the south, especially in the Bohai and Yellow Seas. Based on a 58.1 kg individual consuming 29 g of bivalve molluscs per day, the values of the estimated daily intake (EDI) of trace elements analyzed were significantly lower than the values of the accepted daily intake (ADI) established by Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives (JFAO/WHO) and the guidelines of the reference does (RfD) established by the United States Environmental Protection Agency (USEPA). Additionally, the risk of trace elements to humans through R. philippinarum consumption was also assessed. The calculated hazard quotients (HQ) of all trace elements were less than 1. Consequently, there was no obvious public risk from the intake of these trace elements through R. philippinarum consumption.

Fertilization, Survival, and Growth of Crassostrea hongkongensis ♀ × Crassostrea ariakensis ♂ Hybrids in Northern China

ABSTRACT Hybrids between Crassostrea hongkongensis and Crassostrea ariakensis were produced to evaluate their aquaculture potential in northern China. Fertilization success was asymmetrical; C. hongkongensis eggs can be fertilized by C. ariakensis sperm, but not vice versa. Molecular genetics analysis revealed that hybrid progeny were true hybrids. Larval growth of hybrids was intermediate between C. hongkongensis and C. ariakensis, whereas larval survival in hybrid families ranged from 0.3–37%. The progeny of C. hongkongensis did not survive during an outdoor overwinter phase, whereas hybrids and C. ariakensis showed similar survival rates (>72%). Mean shell height of hybrid adults at day 360 was 54% larger than that of C. hongkongensis adults, and was 25% larger than that of C. ariakensis adults. Morphology of the hybrid adult was similar to that of C. hongkongensis. On the basis of high overwinter survival and faster growth than either parental species, we suggest that the hybrid between C. hongkongensis and C. ariakensis may substitute for C. hongkongensis as a cultured species in northern China.

Biomagnification of trace elements in a benthic food web: the case study of Deer Island (Northern Yellow Sea)

In this study, we investigated the concentrations of ten trace elements (Cu, Zn, Mn, Se, Ni, Cd, Cr, Pb, Hg, and As) and their trophodynamics in a benthic food chain of Deer Island, Northern Yellow Sea. The concentrations of Cu, Zn, Mn, Se, Ni, Cd, Cr, Pb, Hg, and As in the food chain ranged from 3.2 to 23.2, from 71 to 227, from 7.4 to 45.6, from 0.44 to 5.80, from 0.73 to 7.60, from 0.14 to 1.65, from 0.68 to 6.70, from 0.08 to 1.86, from 0.08 to 1.18, and from 0.24 to 3.92 mg kg−1 dry weight, respectively. Among these trace elements, the linear regression between the log-transformed concentrations of Hg and Cd and δ15N values showed statistically significant increase (p<0.05) with the slopes of 0.134 and 0.144, indicating biomagnification of Hg and Cd occurred in the benthic food chain of Deer Island. While the linear regression for other eight trace elements (Cu, Zn, Mn, Se, Ni, Cr, Pb and As) were characterised by extensive scatter with non-significant correlation coefficients (R 2=0.002–0.235) and slopes (p=0.079–0.875), indicating there were not biomagnified or biodiluted of these trace elements.

Identification and characterization of a novel phage-type like lysozyme from Manila clam, Ruditapes philippinarum

A novel lysozyme gene (RpLysPh) with high similarity to the bacteriophage lysozymes was identified in Manila clam, Ruditapes philippinarum. The full length cDNA of RpLysPh is 828bp and contains a 462bp open reading frame (ORF) that codes for a 154 amino acid protein. Multiple sequence alignment analysis revealed that the three residues essential for catalytic activity in phage-type lysozyme (Glu(20), Asp(29), and Thr(35)) are conserved in RpLysPh. The comparison of the 3D models of RpLysPh and Coxiella burnetii lysozyme also suggested that the active sites involved in the binding of substrate have similar conformations. Phylogenetic analysis suggested that RpLysPh shares a similar origin with the bacterial phage-type lysozyme group. The highest level of expression of RpLysPh was observed in hemocytes, followed by mantle. Induction of RpLysPh expression was observed in gills in response to lipopolysaccharide (LPS), peptidoglycan (PGN), polyinosinic-polycytidylic acid (Poly(I:C)), and whole glucan particles (WGP) challenge. The recombinant protein of RpLysPh showed antibacterial activity against both Gram-positive and Gram-negative bacteria.

Concentrations of Selected Trace Elements in Marine Bivalves and the Dietary Risk to Residents of Dalian City, Northern China

ABSTRACT To assess the potential health risks associated with selected trace elements due to consumption of bivalves, a screening-level risk assessment was conducted through consideration of internationally accepted dietary guidelines and the calculation of hazard quotients. Seventeen edible bivalve species were collected from a local market in Dalian City, China. The ranges obtained for the trace elements analyzed in mg/kg dry weight were as follows: Cu (1.8–133), Zn (56–693), Mn (5.7–150), Se (0.2–3.8), Ni (0.56–7.67), Cd (0.04–10.3), Cr (0.48–2.23), Pb (0.01–1.63), Hg (0.07–0.35), and As (0.22–4.00). The estimated daily intake (EDI) of bivalves was 15.0–255.1 times lower than the RfD guidelines for all the trace elements studied. Further, the calculated hazard quotients (HQs) of the trace elements analyzed were well less than 1. Therefore, there were no potential human health risks to Dalian City consumers of marine bivalves.

Transcriptomic analysis of Crassostrea sikamea × Crassostrea angulata hybrids in response to low salinity stress.

Hybrid oysters often show heterosis in growth rate, weight, survival and adaptability to extremes of salinity. Oysters have also been used as model organisms to study the evolution of host-defense system. To gain comprehensive knowledge about various physiological processes in hybrid oysters under low salinity stress, we performed transcriptomic analysis of gill tissue of Crassostrea sikamea ♀ × Crassostrea angulata♂ hybrid using the deep-sequencing platform Illumina HiSeq. We exploited the high-throughput technique to delineate differentially expressed genes (DEGs) in oysters maintained in hypotonic conditions. A total of 199,391 high quality unigenes, with average length of 644 bp, were generated. Of these 35 and 31 genes showed up- and down-regulation, respectively. Functional categorization and pathway analysis of these DEGs revealed enrichment for immune mechanism, apoptosis, energy metabolism and osmoregulation under low salinity stress. The expression patterns of 41 DEGs in hybrids and their parental species were further analyzed by quantitative real-time PCR (qRT-PCR). This study will serve as a platform for subsequent gene expression analysis regarding environmental stress. Our findings will also provide valuable information about gene expression to better understand the immune mechanism, apoptosis, energy metabolism and osmoregulation in hybrid oysters under low salinity stress.

Construction of a High-Density Genetic Map and Quantitative Trait Locus Mapping in the Manila clam Ruditapes philippinarum

Genetic linkage maps are indispensable tools in a wide range of genetic and genomic research. With the advancement of genotyping-by-sequencing (GBS) methods, the construction of a high-density linkage maps has become achievable in marine organisms lacking sufficient genomic resources, such as mollusks. In this study, high-density linkage map was constructed for an ecologically and commercially important clam species, Ruditapes philippinarum. For the consensus linkage map, a total of 9658 markers spanning 1926.98 cM were mapped to 18 sex-averaged linkage groups, with an average marker distance of 0.42 cM. Based on the high-density linkage map, ten QTLs for growth-related traits and shell color were detected. The coverage and density of the current map are sufficient for us to effectively detect QTL for segregating traits, and two QTL positions were all coincident with the closest markers. This high-density genetic linkage map reveals basic genomic architecture and will be useful for comparative genomics research, genome assembly and genetic improvement of R. philippinarum and other bivalve molluscan species.

Interspecific hybridization between Crassostrea angulata and C. ariakensis

Interspecific hybridization can generate heterosis, which is proven to be a useful tool in selective breeding programs for oyster culture. Crassostrea angulata and C. ariakensis are two important economic shellfish species in China. We conducted 2 × 2 reciprocal crosses to determine whether these two species can cross-fertilize and their hybrids can hatch, survive and perform heterosis. Fertilization was found symmetrical without delay. The rate of fertilization success of C. angulata ♀ × C. ariakensis ♂ was lower than that of C. ariakensis ♀ × C. angulata ♂, and the success rate of both hybridizations was lower than that of two intraspecific crosses each. During the planktonic period, survival rate of the progeny was lower in the hybrid crosses than in the intraspecific crosses. On day 360, mean shell height of the progeny of C. angulata ♀ × C. angulata ♂ was highest, which was followed by that of C. angulata ♀ × C. ariakensis ♂, C. ariakensis ♀ × C. ariakensis ♂ and C. ariakensis ♀ × C. angulata ♂ in a descending order. Morphology of adults produced by the hybrid crosses was similar to that of C. angulata. Both hybrids underwent normal gonad development and produced mature gametes in the mating season. This study provided new insights into the quantitative traits in interspecific crosses of Crassostrea species, thus being of guidance value for selective breeding of oyster.