Effects of water depth and substrate color on the growth and body color of the red sea cucumber,Apostichopus japonicus

Three color variants of the sea cucumber,Apostichopus japonicus are recognized,the red one is highly valued in the market. When the red variant is cultured in ponds in China,its body color changes from red to celadon in 3–6 months. The effects of water depth and substrate color on the growth and body color of this animal were investigated. Juveniles of red A. japonicus were cultured in cages suspended at a range of water depths(20,50,100,150 and 200 cm). The specific growth rate of red sea cucumbers was significantly higher in animals cultured at deeper water layers compared with those grown at shallowers. Body weights were greatest for sea cucumbers cultured at a depth of 150 cm and their survival rates were highest at a depth of 200 cm. A scale to evaluate the color of red sea cucumbers(R value) was developed using a Pantone standard color card. All stocked animals in the 9-month trial retained a red color,however the red body color was much more intense in sea cucumbers cultured at shallower depths,while animals suspended in deeper layers became pale. In a separate trial,A. japonicus were cultured in suspended cages with seven different colored substrates. Substrate color had a significant effect on the growth and body-color of red A. japonicus. The yield were greatest for A. japonicus cultured on a yellow substrate,followed by green > white > orange > red > black and blue. All sea cucumbers in the 7-month trial retained a red color,although the red was most intense(highest R value) in animals cultured on a blue substrate and pale(lowest R value) for animals cultured on a green substrate.

Shallow Genetic Structure of Pholis fangi in Bohai Sea and Yellow Sea Inferred from mt DNA Control Region

Pholis fangi is an ecologically important fish species inhabiting Chinese coastal waters of Yellow Sea and Bohai Sea. To investigate the genetic diversity and population genetic structure of P. fangi, a fragment of 487 bp in the first hypervariable region (HVR-1) of the mitochondrial DNA (mtDNA) control region was sequenced for 181 individuals collected from Bohai Sea and Yel- low Sea. A total of 18 polymorphic sites were detected, which defined 25 haplotypes. A moderate level of haplotype diversity (h = 0.7052) and a low level of nucleotide diversity (π= 0.0028) were detected. Both the phylogenetic tree and the haplotype network showed no significant genealogical structure difference among sampling locations. Pairwise FST comparison and hierarchical mo- lecular variance analysis (AMOVA) revealed that no significant genetic structure difference existed throughout the investigated re- gions, suggesting a high gene exchange among different populations. The results of neutrality test and mismatch distribution analysis indicated that a late Pleistocene population expansion (38000 127000 years ago) happened. Seasonal schooling migration may con- tribute to the genetically homogeneous population structure of the species.

Integrated transcriptome and physiology analysis of Microcystis aeruginosa after exposure to copper sulfate

Copper sulfate(CuSO4)is widely used in controlling the Microcystis aeruginosa blooms.Many studies reported the toxicity mechanisms of Cu^2+to M.aeruginosa at the physiological level,but little is known about a transcriptomic basis of these mechanisms.In the present study,M.aeruginosa was treated by 0.5 mg/L Cu^2+(half of the 96-h EC50)for 72 h.The results show that CH2+content in M.aeruginosa increased after 72 h Cu^2+exposure,whereas the Fv/Fm chlorophyll fluorescence value and chlorophyll a content in M.aeruginosa sharply decreased.Reactive oxygen species concentration and activity of antioxidant enzymes(superoxide dismutase,catalase and peroxidase)were all increased.These physiological data confirmed toxicity of Cu^2+to m.aeruginosa.The RNA-seq analysis showed that 6646725 and 7880291 clean reads were obtained for the Cu-treated and control libraries,respectively.The 595 genes(252 downward trend and 343 upward trend)with the Gene Ontology(GO)annotations were divided into three main functional categories:cellular component,molecular function,and biological process.In the Cluster of Orthologous Groups(COG)annotation,418 differentially expressed genes with 25 functional definitions were obtained.Among them,‘replication,recombination and repair’,‘energy production and conversion’,and‘general function prediction only’were the largest three groups of transcripts.In the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,genes involved in photosynthesis and oxidative phosphorylation were present at the highest percentages.In addition,the genes involved in photosynthesis and oxidative phosphorylation were identified,and then confirmed using real-time PCR.This study reported the first transcriptome of M.aeruginosa.Photosynthesis and oxidative phosphorylation were severely affected by Cu^2+toxicity,which may have contributed to cell death.These data provide the potential mechanism to explain the CuSO4 effect on the harmful M.aeruginosa.

Carbon and nitrogen budget in fish-polychaete integrated aquaculture system

Integrated multi-tropic aquaculture(IMTA)systems have been used in China for many years and have achieved significant economic,social,and ecological benefits.However,there is still a lack of benthic bioremediation species that can effectively utilize the aquaculture particulate organic waste in the system.Polychaete Perinereis aibuhitensis Grube is used as an environmental remediation species for large-scale aquaculture to reduce particulate organic waste,which is of great significance to environmental protection.To improve bio-elements utilization efficiency,P.aibuhitensis was applied for IMTA indoor fish(Hexagrammos otakii)farming.Results showed that in the system,production of 1 kg of the fish discharged 2141-2338 mg of carbon and 529-532 mg of nitrogen,while in the monoculture of the fish,the figures were 3033-3390 mg and 764-794 mg,or 24.84%-35.26%and 30.35%-33.32%less,respectively.This approach promoted IMTA technology that could utilize the particulate organic waste from intensive aquaculture and reduce the adverse environmental effects.

Molecular Characterization and Expression and DNA Methylation Analyses of a Galectin-Related Protein Gene from Cynoglossus semilaevis

Galectins, a family of β-galactoside-binding proteins, participate in both innate immunity and adaptive immunity. This study identified one novel galectin-related protein from half-smooth tongue sole Cynoglossus semilaevis, which was designated as CsGRP. The full-length cDNA of its encoding gene was 785 bp in length with a 528 bp open reading frame encoding a putative protein of 176 amino acids. The deduced CsGRP contained a putative 131-aa galactoside-binding lectin domain and 3 critical residues responsible for carbohydrate binding （R93, W109 and R114）. Genomic structural analysis revealed that CsGRP consisted of five exons and four intzons. CsGRP showed 68% similarity with Poecilia latipinna GRP and 67% similarity wih Stegastes partitus GRP. CsGRP showed the highest expression level in liver, although its expression was detected in all tested tissues. When challenged with Vibrio harveyi, the expression of CsGRP was significantly down-regulated in liver （P〈 0.05）. In addition, we found that in spleen and kidney of C. semilaevis, the CpG island of CsGRP showed significantly higher （P 〈 0.05） methylation level in disease-resistant family of C. semilaevis （DR-Cs） than in disease-susceptible ones （DS-Cs）. Our results suggested that CsGRP may play important roles in the immune response of C. semilaevis. Moreover, DNA methylation patterns provided valuable data for understanding the relationship between epigenetic regulation and immunity, which would assist the animal genetic research and improve the animal breeding in the future.

Air Temperature and Emersion Time Can Affect the Survival Rate and Ammonium Loading of Swimming Crab Portunus trituberculatus Exposed to Air

Ammonium overloading is a common response of aquatic organisms to air exposure during transport. This study elucidated the relationship between ammonium overloading and mortality of crab Portunus trituberculatus. Additionally, we also explored the effects of emersion time and air temperature on ammonium loading and concomitant physiological change. To test the air temperature effect, the crab was exposed to 16, 23 and 30℃ in air for 3 h, respectively, and then recovered in seawater at 23℃ for 12 h. To test the emersion time effect, crab was exposed to 23℃ in air for 0.5 and 3 h, respectively, and then recovered in seawater at 23℃ for 12 h. In the control group, crab was always immersed at 23℃. At each time interval(0.5, 1.5 and 3 h during air exposure and 0.5, 2, 4 and 12 h during recovery), ammonium excretion rate, level of total ammonium, total free amino acids and urea concentration in hemolymph and the hepatopancreas enzyme activity involved in detoxifying ammonium were analysed. Results showed that crab mortality was positively related with emersion time and temperature while ammonium loading was lower at 16 and 30℃ than at 23℃. For crab experiencing thermal inconsistence of culture media(i.e., 16 or 30℃), they were higher in ammonium excretion rate and activities of ammonium detoxification enzymes, which may be the reason that they had a lower ammonium loading. Prolonged emersion time(3.0 h vs. 0.5 h) increased the ammonium overloading and the activity of ammonium detoxification pathways in crab. Our results demonstrated that emersion-induced ammonium overloading may not be the main reason leading to P. trituberculatus death during air exposure and subsequent recovery. When the culture medium changed, thermal variation, compared with constant temperature, could reduce ammonium overloading in crab by elevating the activities of ammonium detoxification enzymes and ammonium excretion rate during recovery period.

Effect of polysaccharide from Enteromorpha prolifera on maize seedlings under NaCl stress

In this study, a polysaccharide from Enteromorpha prolifera (EP) was extracted and its effect on maize seedlings under NaCl stress was investigated. Firstly, the components and structure of the EP were determined. We found that EP is a sulfated polysaccharide of high-molecular weight (Mw, 1 840 KDa) heteropolysaccharides and the main monosaccharide is rhamnose. The polysaccharide was applied to explore its effect on the growth of maize seedlings and its defense response under a salt stress. The results show that EP could promote the growth of maize seedlings under the salt stress. In addition, EP was shown able to significantly regulate membrane permeability and adjustment of osmotic substances such as soluble protein, soluble sugar, and proline, antioxidant enzymes containing superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase. Therefore, EP is an effective salt-resistant substance for the growth of maize seedlings under NaCl stress.

Characterization of Genome-Wide Microsatellites of Saccharina japonica Based on a Preliminary Assembly of Illumina Sequencing Reads

Microsatellites or simple sequence repeats(SSR) function widely and locate dependently in genome. However, their characteristics are often ignored due to the lack of genomic sequences of most species. Kelp(Saccharina japonica), a brown macroalga, is extensively cultured in China. In this study, the genome of S. japonica was surveyed using an Illumina sequencing platform, and its microsatellites were characterized. The preliminarily assembled genome was 469.4 Mb in size, with a scaffold N_(50) of 20529 bp. Among the 128370 identified microsatellites, 90671, 25726 and 11973 were found in intergenic regions, introns and exons, averaging 339.3, 178.8 and 205.4 microsatellites per Mb, respectively. These microsatellites distributed unevenly in S. japonica genome. Mononucleotide motifs were the most abundant in the genome, while trinucleotide ones were the most prevalent in exons. The microsatellite abundance decreased significantly with the increase of motif repeat numbers, and the microsatellites with a small number of repeats accounted for a higher proportion of the exons than those of the intergenic regions and introns. C/G-rich motifs were more common in exons than in intergenic regions and introns. These characteristics of microsatellites in S. japonica genome may associate with their functions, and ultimately their adaptation and evolution. Among the 120140 pairs of designed microsatellite primers, approximately 75% were predicted to be able to amplify S. japonica DNA. These microsatellite markers will be extremely useful for the genetic breeding and population evolution studies of kelp.

Adsorption characteristics of chitooligosaccharides onto activated charcoal in aqueous solutions

To investigate the adsorption characteristics of chitooligosaccharides in solution onto activated charcoal,we studied the optimal adsorption conditions and the adsorption mechanisms of the chitooligosaccharides onto activated charcoal,which will greatly promote the application of activated charcoal in the chitooligosaccharides separation and purification.We studied the effects of particle size of activated charcoal,pH of solution,contact time,temperature,and initial concentration of chitooligosaccharides on the adsorption behavior in batch mode experiments.Activated charcoal in fine particle size showed a high uptake of chitooligosaccharides.Weak alkaline solution(pH 8–9)was the most favorable to the adsorption.The adsorption equilibrium after 60 min was established,which followed a pseudo-second-order kinetic model.The adsorption capacity(Q_(max))reached 0.195 g/g(chitooligosaccharides/activated charcoal)at 298 K.The adsorption was temperature-insensitive,and the adsorption isotherms could be best described by the Langmuir equation.Chitooligosaccharides adsorbed on activated charcoal could be desorbed in 50%ethanol solution in combination with an acidic condition(pH 2),reaching desorption efficiency of 96.0%.These findings are of great significance for the production and purification of amino oligosaccharides including chitooligosaccharides using activated charcoal.

Ecological effects of co-culturing the sea cucumber Apostichopus japonicus with the Chinese white shrimp Fenneropenaeus chinensis in an earthen pond

Using net enclosures in an earthen pond, we established three culture treatments with the sea cucumber Apostichopus japonicus and the Chinese white shrimp Fenneropenaeus chinensis : monoculture of sea cucumbers(C), monoculture of shrimp(S), and co-culture of the two species(CS). We measured levels of suspended particulate matter in the water column; total organic matter, total organic carbon, total nitrogen, and carbon/nitrogen ratios in both settling particles and the sediment; and chlorophyll a levels in the sediment. We then compared these variables between the three treatments. We also examined growth, survival, and yield of the two species in the different treatments. From June to September, the mean monthly suspended particulate matter sedimentation rates in the CS and S treatments were significantly( P <0.05) greater than those in the C treatment. From August to November, the mean monthly total organic matter, total organic carbon, total nitrogen, and chlorophyll a contents in the sediment in the CS and S treatments were significantly( P <0.05) greater than those in the C treatment. Final wet weight, specific growth rate, survival rate, and total yield of sea cucumbers in co-culture were all significantly greater than those of sea cucumbers in monoculture. There were no significant differences among any of these variables for shrimp reared in the two systems. The bioturbation of the sediment and fecal production of the shrimp likely supplied natural food for the sea cucumbers. Co-culture of the two species is a viable option for increasing yield per unit area, maximizing use of the water body, and diversifying crop production.

Environmental significance of biogenic elements in surface sediments of the Changjiang Estuary and its adjacent areas

Biogenic elements and six phosphorus （P） fractions in surface sediments from the Changjiang Estuary and adjacent waters were determined to investigate the governing factors of these elements, and further to discuss their potential uses as paleo-environment proxies and risks of P release from sediment. Total organic carbon （TOC） and leachable organic P （Lea-OP） showed high concentrations in the estuary, Zhejiang coast and offshore upwelling area. They came from both the Changjiang River and marine biological input. Biogenic silicon （BSi） exhibited a high concentration band between 123 and 124°E. BSi mainly came from diatom production and its concentration in the inshore area was diluted by river sediment. Total nitrogen （TN） was primarily of marine biogenic origin. Seaward decreasing trends of Fe-bound P and Al-bound P revealed their terrestrial origins. Influenced by old Huanghe sediment delivered by the Jiangsu coastal current, the maximum concentration of detrital P （Det-P） was observed in the area north of the estuary. Similar high concentrations of carbonate fluorapatite （CFA-P） and CaCO3in the southern study area suggested marine calcium-organism sources of CFA-P. TOC, TN and non-apatite P were enriched in fine sediment, and Det-P partially exhibited coarse-grain enrichment, but BSi had no correlation with sediment grain size. Different sources and governing factors made biogenic elements and P species have distinct potential uses in indicating environmental conditions. Transferable P accounted for 14%-46% of total P. In an aerobic environment, there was low risk of P release from sediment, attributed to excess Fe oxides in sediments.

High-resolution linkage and quantitative trait locus mapping using an interspecific cross between Argopecten irradians irradians(♀) and A. purpuratus (♂)

The bay scallop and Peruvian scallop are economically important species.Interspecific hybrids of these two scallops outperformed both of their parent species in multiple growth traits but exhibited decreased fertility,which provides good models for the study of heterosis and species divergence.Genetic mapping serves as a chromosomal-level framework to investigate the molecular mechanisms of hybridization and introgression.In this study,high-resolution linkage maps were constructed for the bay and Peruvian scallops with an interspecific hybrid family.The linkage map of the bay scallop covered over 98.9% of the whole genome with 2994 mapped markers and the average marker interval of 0.32 cM.For the Peruvian scallop,1585 markers were mapped with the average maker interval of 0.51 cM,covering 97.7% of the genome.Both the two linkage maps have 16 linkage groups,corresponding to the haploid chromosome number of the two species.Approximately,54.5% of markers exhibited significant deviation from the expected Mendelian ratio of segregation,lending in sights into the intrinsic incompatibility between the two species.QTLs related to growth and shell coloration were detected,which could explain 13.1%and 74.9% of the phenotypic variance,respectively.This represents important information for further evaluation.These findings are an important addition to the genomic resources for scallop genetic studies,and are especially useful for investigations on genomic incompatibility for hybridization,genome evolution of closely related species,and genetic enhancement programs in aquaculture.