Acute toxicity of live and decomposing green alga Ulva (Enteromorpha) prolifera to abalone Haliotis discus hannai

From 2007 to 2009, large-scale blooms of green algae (the so-called "green tides") occurred every summer in the Yellow Sea, China. In June 2008, huge amounts of floating green algae accumulated along the coast of Qingdao and led to mass mortality of cultured abalone and sea cucumber. However, the mechanism for the mass mortality of cultured animals remains undetermined. This study examined the toxic effects of Ulva (Enteromorpha) prolifera, the causative species of green tides in the Yellow Sea during the last three years. The acute toxicity of fresh culture medium and decomposing algal effluent of U. prolifera to the cultured abalone Haliotis discus hannai were tested. It was found that both fresh culture medium and decomposing algal effluent had toxic effects to abalone, and decomposing algal effluent was more toxic than fresh culture medium. The acute toxicity of decomposing algal effluent could be attributed to the ammonia and sulfide presented in the effluent, as well as the hypoxia caused by the decomposition process.

Heterosis and combining ability:a diallel cross of three geographically isolated populations of Pacific abalone Haliotis discus hannai Ino

We conducted a complete diallel cross among three geographically isolated populations of Pacific abalone Haliotis discus hannai Ino to determine the heterosis and the combining ability of growth traits at the spat stage.The three populations were collected from Qingdao(Q) and Dalian(D) in China,and Miyagi(M) in Japan.We measured the shell length,shell width,and total weight.The magnitude of the general combining ability(GCA) variance was more pronounced than the specific combining ability(SCA) variance,which is evidenced by both the ratio of the genetic component in total variation and the GCA/SCA values.The component variances of GCA and SCA were significant for all three traits(P<0.05),indicating the importance of additive and non-additive genetic effects in determining the expression of these traits.The reciprocal maternal effects(RE) were also significant for these traits(P<0.05).Our results suggest that population D was the best general combiner in breeding programs to improve growth traits.The DM cross had the highest heterosis values for all three traits.

Acclimation-dependent expression of heat shock protein 70 in Pacific abalone (Haliotis discus hannai Ino) and its acute response to thermal exposure

Heat shock protein 70 （Hsp70） is one important member of heat shock protein （Hsp） family that is responsible for various stresses, especially thermal stress. Here we examined the response of Hsp70 gene to both chronic and acute thermal exposure in Pacific abalone （Haliotis discus hannai Ino）. For the chronic exposure, abalones were maintained at 8, 12, 20, and 30∩ for four months and their mRNA levels were measured. The highest mRNA level of Hsp70 gene relative to actin gene was detected in the 30~C-acclimated group, followed by the 8~C-acclimated group and then the 12~C- and 20~C-acclimated groups. After the long-term acclimation, gills from each of the above acclimation groups were dissected and exposed to different temperatures between 8~C and 38~C for 30 min. Hsp70 expression in gills acclimated to different temperatures responded differentially to the same temperature exposure. The incubation temperature that induced maximum Hsp70 mRNA expression was higher in the higher temperature acclimation groups than lower temperature groups. Pacific abalones could alter the expression pattern of Hsp70 gene according to environmental thermal conditions, through which they deal with the stress of thermal variations.

Effects of temperature and salinity on survival, growth and DNA methylation of juvenile Pacific abalone, Haliotis discus hannai Ino

Temperature and salinity are two of the most potent abiotic factors influencing marine mollusks. In this study, we investigated the individual and combined effects of temperature and salinity on the survival and growth of juvenile Pacific abalone, Haliotis discus hannai Ino, and also examined the DNA methylation alteration that may underpin the phenotypic variation of abalone exposed to different rearing conditions. The single-factor data showed that the suitable ranges of temperature and salinity were 16-28℃ at a constant salinity of 32, and 24-40 at a constant temperature of 20℃, respectively. The two-factor data indicated that both survival and growth were significantly affected by temperature, salinity and their interaction. The optimal temperature-salinity combination for juveniles was 23-25℃ and 30-36. To explore environment-induced DNA methylation alteration, the methylation-sensitive amplified polymorphism （MSAP） technique was used to analyze the genomic methylation profiles of abalone reared in optimal and adverse conditions. Neither temperature nor salinity induced evident changes in the global methylation level, but 67 and 63 differentially methylated loci were identified in temperature and salinity treatments, respectively. The between-group eigen analysis also showed that both temperature and salinity could induce epigenetic differentiation in H. discus hannai Ino. The results of our study provide optimal rearing conditions for juvenile tt. discus hannai Ino, and represent the first step toward revealing the epigenetic regulatory mechanism of abalone in response to thermal and salt stresses.

Effects of dietary menadione on the activity of antioxidant enzymes in abalone, Haliotis discus hannai Ino

A 240-day growth experiment in a re-circulating water system was conducted to investigate the effects of dietary menadione on the growth and antioxidant responses of abalone Haliotis discus hannai Ino. Triplicate groups of juvenile abalone (initial weight: 1.19 ± 0.01 g; shell length: 19.23 ± 0.01 mm) were fed to satiation with 3 semi-purified diets containing 0, 10, and 1 000 mg menadione sodium bisulfite (MSB)/kg, respectively. Results show that there were no significant differences in the rate of weight gain or in the daily increment in shell length of abalone among different treatments. Activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione S-transferase (GST) and glutathione reductase (GR) in viscera were significantly decreased with dietary menadione. However, activities of these enzymes except for GPX in muscle were increased. Therefore, antioxidant responses of abalone were increased in muscle and decreased in viscera by dietary menadione.

Heat transfer characteristics of air cross-flow for in-line arrangement of spirally corrugated tube and smooth tube bundles

An experimental study on heat transfer and resistance coefficients of linearly arranged smooth and spirally corrugated tube bundles in cross-flow was performed. The heat transfer and resistance coefficients are presented in this paper with transverse and longitudinal tube-pitch and tube geometries taken into account. The experiment's results can provide technical guidelines for application to horizontal air preheater with arranged in-line spirally corrugated tube bundles, especially to the air preheater for CFBCBs (Circulating Fluidized Bed Combustion Boilers).

Effects of Waterborne Cu and Cd on Anti-oxidative Response, Lipid Peroxidation and Heavy Metals Accumulation in Abalone Haliotis discus hannai Ino

The aim of this study was to compare the effects of waterbome copper （Cu） and cadmium （Cd） on survival, anti-oxida- tive response, lipid peroxidation and metal accumulation in abalone Haliotis discus hannai. Experimental animals （initial weight： 7.49 g±0.01 g） were exposed to graded concentrations of waterborne Cu （0.02, 0.04, 0.06, 0.08 mg L-1） or Cd （0.025, 0.05, 0.25, 0.5 mgL-1） for 28 days, respectively. Activities of the anti-oxidative enzymes （catalase, CAT; superoxide dismutase, SOD; glutathione peroxidases, GPx; glutathione S-transferase, GST）, contents of the reduced glutathione （GSH） and malondiadehyde （MDA） in the hepatopancreas, and metal accumulation in hepatopancreas and muscles were analyzed after 0, 1, 3, 6, 10, 15, 21, 28 days of metal exposure, respectively. Results showed that 0.04 mg L-1, 0.06 mg L-1 and 0.08 mgL-1 Cu caused 100% death of abalone on the 21st, 10th and 6th day, respectively. However, no dead abalone was found during the 28-day waterborne Cd exposure at all experimental concentrations. Generally, activities of SOD and GST in hepatopancreas under all Cu concentrations followed a decrease trend as the exposure time prolonged. However, these activities were firstly increased and then decreased to the control level and increased again during Cd exposure. Activities of CAT in all Cu exposure treatments were higher than those in the control. These activities were firstly increased and then decreased to the control level and increased again during Cd exposure. Contents of MDA in hepatopancreas in all Cu treatments significantly increased first and then decreased to the control level. However, the MDA contents in hepatopan- creas were not significantly changed during the 28-day Cd exposure. The metals accumulation in both hepatopancreas and muscles of abalone significantly increased with the increase of waterborne metals concentration and exposure time. These results indicated that H. discus hannai has a positive anti-oxidative defense against Cu or Cd. In conclusion, anti-oxidative mechanism in abalone to resist waterborne Cu did not follow the same pattern as that for waterborne Cd.

Characterization of genic microsatellite markers derived from expressed sequence tags in Pacific abalone (Haliotis discus hannai)

Simple sequence repeat (SSR) markers were developed from the expressed sequence tags (ESTs) of Pacific abalone (Haliotis discus hannai).Repeat motifs were found in 4.95% of the ESTs at a frequency of one repeat every 10.04 kb of EST sequences,after redundancy elimination.Seventeen polymorphic EST-SSRs were developed.The number of alleles per locus varied from 2-17,with an average of 6.8 alleles per locus.The expected and observed heterozygosities ranged from 0.159 to 0.928 and from 0.132 to 0.922,respectively.Twelve of the 17 loci (70.6%) were successfully amplified in H.diversicolor.Seventeen loci segregated in three families,with three showing the presence of null alleles (17.6%).The adequate level of variability and low frequency of null alleles observed in H.discus hannai,together with the high rate of transportability across Haliotis species,make this set of EST-SSR markers an important tool for comparative mapping,marker-assisted selection,and evolutionary studies,not only in the Pacific abalone,but also in related species.

Identification of normalization factors for quantitative realtime RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai

Quantitative real-time reverse transcription-polymerase chain reaction （qRT-PCR） is widely used in studies of gene expression. In most of these studies, housekeeping genes are used as internal references without validation. To identify appropriate reference genes for qRT-PCR in Pacific abalone Haliotis discus hannai, we examined the transcription stability of six housekeeping genes in abalone tissues in the presence and absence of bacterial infection. For this purpose, abalone were infected with the bacterial pathogen Fibrio anguillarum for 12 h and 48 h. The mRNA levels of the housekeeping genes in five tissues （digestive glands, foot muscle, gill, hemocyte, and mantle） were determined by qRT-PCR. The PCR data was subsequently analyzed with the geNorm and NormFinder algorithms. The results show that in the absence of bacterial infection, elongation factor-l-alpha and beta-actin were the most stably expressed genes in all tissues, and thus are suitable as cross-tissue type normalization factors. However, we did not identify any universal reference genes post infection because the most stable genes varied between tissue types. Furthermore, for most tissues, the optimal reference genes identified by both algorithms at 12 h and 48 h post-infection differed. These results indicate that bacterial infection induced significant changes in the expression of abalone housekeeping genes in a manner that is dependent on tissue type and duration of infection. As a result, different normalization factors must be used for different tissues at different infection points.

Effect of Dietary Lipid on the Growth, Fatty Acid Composition and Δ5 Fads Expression of Abalone(Haliotis discus hannai Ino) Hepatopancreas

This study investigated the effect of dietary lipid on the growth, fatty acid composition and Δ5 fatty acyl desaturase genes(Fads) expression of juvenile abalone(Haliotis discus hannai Ino) hepatopancreas. Six purified diets were formulated to contain tripalmitin(TP), olive oil(OO, 72.87% 18:1n-9), grape seed oil(GO, 68.67% 18:2n-6), linseed oil(LO, 70.48% 18:3n-3), ARA oil(AO, 41.81% ARA) or EPA oil(EO, 44.09% EPA and 23.67% DAH). No significant difference in survival rate was observed among abalone fed with different diets. Weight gain rate(WGR) and daily growth rate of shell length(DGRSL) were significantly increased in abalone fed with diets containing OO, AO and EO, but decreased in abalone fed with LO diet(P < 0.05) in comparison with those fed with TP. High level of dietary 18:2n-6 resulted in higher content of n-6 polyunsaturated fatty acids(PUFAs) in abalone fed with GO than those fed with TP, OO, LO and EO(P < 0.05). n-3 PUFAs in abalone fed with LO was significantly higher than those in abalone fed with TP, OO, GO and AO(P < 0.05). The highest contents of 20:1n-9 and 22:1n-9 were observed in abalone fed with OO. The expression of Δ5 Fads in hepatopancreas of abalone was enhanced by high concentration of 18:3n-3 and suppressed by dietary LC-PUFAs; however it was not affected by dietary high concentration of 18:1n-9 or 18:2n-6. These results provided valuable information for understanding the synthesis of LC-PUFAs and nutritional regulation of Δ5 Fads expression in abalone.

Wrinkling and Growth Mechanism of CuO Nanowires in Thermal Oxidation of Copper Foil

We report a systematic study on wrinkling and CuO nanowires （NWs） growth in the thermal oxidation of copper foil. Copper foils with thickness of 0.5 mm were thermally oxidized in air at 500℃ for 0.5-10 h. It is found that all the samples have wrinkles and the size of the wrinkles increases with the oxidation time increasing. CuO NWs can grow on both the sidehill and hilltop of wrinkle. The CuO NWs on sidehill are longer and denser than those on hilltop. The growth direction of the CuO NWs on sidehill is not vertical to the substrate but vertical to their growth surfaces. The process of wrinkling and CuO NWs growth can be divided into three stages： undulating, voiding, and cracking. The CuO NWs on both sidehill and hilltop grow at the undulating stage. However, only the CuO NWs on sidehill grow and those on hilltop stop growing at the voiding and cracking stages because of the void in hilltop. The local electric field in a wrinkle at undulating stage was calculated, and it is found that the difference of local electric field strengths between hilltop and sidehill is small, which indicates that the predominant driving force for the diffusion of Cu ion during CuO NWs growth is internal stress.

Dietary guaiacol improves the growth of juvenile abalone,Haliotis discus hannai Ino

We conducted a preliminarily study on the effects of dietary guaiacol on growth performance of abalone, Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol （0, 5, 10, 50, 100, 500, and 1 000 mg kgl）. Abalone （initial weight： 0.29 ± 0.01 g; initial shell length： 8.55 ± 0.27 mm） were fed with these diets in a re-circulated water system for 152 days. Guaiacol significantly improved the specific growth rate （SGR） （P〈0.05）; excessive dietary guaiacol （1 000 mg kg1） led to significantly high mortality （P〈0.05）, and lipid content in the soft body increased significantly after dietary guaiacol （P〈0.05）. Activities of catalase and phenoloxidase （PO） in the viscera were significantly stimulated by dietary guaiacol （P〈0.05）. Broken-line analysis based on SGR indicated that the minimum dietary guaiacol for the optimal growth of juvenile abalone is 15.43 mg ·kg^-1.

Effects of Dietary Carbohydrates Sources on Lipids Compositions in Abalone, Haliotis discus hannai Ino

A study was conducted to evaluate the effects of dietary carbohydrates on triglyceride, cholesterol and fatty acid concentrations in abalone, Haliotis discus hannai Ino. Six semi-purified diets with different carbohydrates （dextrin, heat-treated wheat starch, wheat starch, corn starch, tapioca starch and potato starch, respectively）, all containing a carbohydrate level of 33.5%, were fed to abalone （initial shell length： 29.98mm±0.09mm; initial weight. 3.42g±0.02g） for 24 weeks in a recirculation system. The results indicate that serum triglyceride concentrations were significantly （P〈0.05） higher in the abalone fed with dextrin, heat-treated wheat starch and wheat starch than those fed with corn starch, and serum cholesterol concentrations were significantly （P〈0.05） higher in the abalone fed with dextrin, heat-treated wheat starch than those fed with corn starch. Fatty acid C20：4n-6 in the foot mus- cles were significantly （P〈0.05） lower in the abalone fed with dextrin than those fed with wheat starch, corn starch, tapioca starch and potato starch. Fatty acid C20：4n-6 in hepatopancreas was significantly （P〈0.05） lower in abalone fed with heat-treated wheat starch than those fed with corn starch, tapioca starch and potato starch. Fatty acid C22：6n-3 in the foot muscles were significantly （P 〈 0.05） lower in the abalone fed with dextrin and beat-treated wheat starch than those fed with wheat starch and potato starch.

Fiber Bragg Grating Pressure Sensor Based on Corrugated Diaphragm

A kind of fiber Bragg grating pressure sensor based on corrugated diaphragm is proposed. The relationship between the central wavelength of reflective wave of FBG and pressure is given, and the expression of the pressure sensitivity coefficient is also given. Within the range from 0 MPa to 0.3 MPa, the experimental pressure sensitivity is 7.83 nm/MPa, which is 2 610 times than that of the bare fiber grating. The experimental results agree with the theoretical analysis. It is indicated that the expected pressure sensitivity of the sensor can be obtained by optimizing the size and mechanical parameters of the corrugated diaphragm.

The Origin of Wrinkles on Transferred Graphene

When two-dimensional graphene is exfoliated from three-dimensional highly oriented pyrolytic graphite （HOPG）, ripples or corrugations always exist due to the intrinsic thermal fluctuations. Surface-grown graphenes also exhibit wrinkles, which are larger in dimension and are thought to be caused by the difference in thermal expansion coefficients between graphene and the underlying substrate in the cooling process after high temperature growth. For further characterization and applications, it is necessary to transfer the surface-grown graphenes onto dielectric substrates, and other wrinkles are generated during this process. Here, we focus on the wrinkles of transferred graphene and demonstrate that the surface morphology of the growth substrate is the origin of the new wrinkles which arise in the surface-to-surface transfer process; we call these morphology- induced wrinkles. Based on a careful statistical analysis of thousands of atomic force microscopy （AFM） topographic data, we have concluded that these wrinkles on transferred few-layer graphene （typically 1-3 layers） are determined by both the growth substrate morphology and the transfer process. Depending on the transfer medium and conditions, most of the wrinkles can be either released or preserved. Our work suggests a new route for graphene engineering involving structuring the growth substrate and tailoring the transfer process.

Crystal-oriented wrinkles with origami-type junctions in few-layer hexagonal boron nitride

Understanding layer interplay is the key to utilizing layered heterostructures formed by the stacking of different two-dimensional materials for device applications. Boron nitride has been demonstrated to be an ideal substrate on which to build graphene devices with improved mobilities. Here we present studies on the morphology and optical response of annealed few-layer hexagonal boron nitride flakes deposited on a silicon substrate that reveal the formation of linear wrinkles along well-defined crystallographic directions. The wrinkles formed a network of primarily threefold and occasionally fourfold origami-type junctions throughout the sample, and all threefold junctions and wrinkles formed along the armchair crystallographic direction. Furthermore, molecular dynamics simulations yielded, through spontaneous symmetry breaking, wrinkle junction morphologies that are consistent with both the experimental results and the proposed origami-folding model. Our findings indicate that this morphology may be a general feature of several two-dimensional materials under proper stress-strain conditions, resulting in direct consequences in device strain engineering.

Regularity analysis of wrinkles under the action of capillary force in an annular thin film

The wrinkling law of annular sheet which is induced by capillary force with inner liquid film is analyzed in this paper.The results show that the inner liquid film can wrinkle the annular sheet when the surface tension of the liquid film reaches a critical value,and the critical value can be dramatically altered by changing the geometry and properties of the annular sheet.The results obtained in this article may hold potential applications in generating three-dimensional structures through capillary effects.