Liqiang Zhao

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.

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.

Larval and juvenile growth performance of Manila clam hybrids of two full-sib families

In order to determine whether growth performance could be improved by hybridizing full-sib families of Manila clam (Ruditapes philippinarum), crosses between two full-sib families including self and reciprocal crosses were carried out. The effects of heterosis, combining ability and interaction on the growth of shell length were estimated. The results showed that the growth of hybrid larvae was intermediate between parents on days 6 and 9. Heterosis on shell length was observed, which varied at juvenile stage. The cross of ♂A × ♀B (Hp varied between 10.41% and 68.27%) displayed larger heterosis than ♂B × ♀A (Hp varied between 1.89% and 32.33%) did, suggesting that ♂A × ♀B was an ideal hatchery method of improving the growth performance of Manila clam. The variances of general combining ability (GCA), special combining ability (SCA) and interaction (I) were significant in shell length (P < 0.05), indicating that both additive and non-additive genetic factors were important contributors to the growth of larvae and juveniles. The GCA for shell length of ♂A × ♀B was higher than that of ♂B × ♀A at both larval and juvenile stages. This confirmed that the cross between ♂A and ♀B showed great growth in shell length. In summary, the growth of Manila clam seeds could be improved by hybridizing selected parents from large numbers of full-sib families.

Development and characterization of EST-derived microsatellite makers for Manila clam ( Ruditapes philippinarum )

Abstract Twenty-five polymorphic microsatellite makers were developed and characterized from expressed sequence tag sequence of the Manila clam, Ruditapes philippinarum. The number of alleles at each locus ranged from 3 to 20 with an average of seven alleles per locus. The observed and expected heterozygosity varied from 0.081 to 0.730 and from 0.127 to 0.926, respectively. Thirteen loci were found deviate significantly from Hardy–Weinberg equilibrium. These microsatellite loci will be useful for further studies on the population structure and genetic variation of this species.

Seasonal Variations in Biochemical Composition of the Clam Dosinia corrugate in Relation to the Reproductive Cycle and Environmental Conditions

ABSTRACT Seasonal variations in biochemical composition of the clam Dosinia corrugate were investigated from August 2013 until July 2014 in Zhuanghe Bay in relation to environmental conditions and reproductive cycle. Separate biochemical analyses of the mantle, adductor muscle, foot, and gonad-visceral mass in each sex were undertaken. Spawning took place once a year from July to August and the massive spawning occurred in August with the highest water temperature and chlorophyll a levels. Utilization of glycogen took place during the spawning period, whereas protein was biosynthesized as the mature gametes formed. The glycogen value increased during the resting stage (autumn—winter). The reproductive pattern exhibited by D. corrugate based on the storage of reserves during the period of sexual rest, and the period of gonad maturation, seems to be conservative and opportunistic. The information obtained in this study provides a basis for fisheries resource administrators to design biologically sound management measures for this species, and is crucial for initiating its aquaculture.

Correlation and path analysis of morphological and weight traits in marine gastropod Glossaulax reiniana

Using correlation and path analysis, the genetic correlation between weight traits and morphological traits was determined in the marine gastropod Glossaulax reiniana. A total of 100 G. reiniana individuals from a wild population were used. Shell width (X1), shell height (X2), umbo-callus height (X3), body width (X4), operculum length (X5), operculum width (X6), body weight (Y1) and soft-tissue weight (Y2) were measured, and the correlation coefficient matrix calculated. Morphological traits were used as independent variables and weight traits as dependent variables for path coefficient analysis. Path coefficients, correlation indices and determination coefficients were also determined. Results indicate that the correlation coefficients associated with each morphological and weight trait were all highly significant (P <0.01). After deleting redundant independent variables, the following optimum multiple regression equations were obtained using stepwise multiple regression analysis: Y1 = − 29.317+0.362 X2 +0.349 X4 + 1.190 X5 for body weight; and Y2 = − 17.292+0.166 X1 +0.171 X2 +0.703 X5, for soft-tissue weight. Operculum height had the highest positive direct correlation with both body weight and soft-tissue weight, which was in accordance with the test results obtained from determinate coefficient analysis. The indication of high genetic correlations between weight traits and morphological traits will provide valuable information for G. reiniana breeding programs.