Molecular Characterization and Antibacterial Activity of a G-Type Lysozyme in Yellow Drum(Nibea albiflora)

Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genome-wide association analysis,the g-type lysozyme gene,which is named NaLyg in yellow drum(Nibea albiflora),was found to be a key candidate gene for disease resistance in response to Vibrio harveyi infection.The cDNA of NaLyg was 1025 bp,including four exons and three introns,and its open reading frame(ORF)had a full-length of 582 bp,encoding 193 amino acids.NaLyg was found to be conserved during evolution through bioinformatic analyses.The NaLyg protein possessed a sugar binding domain and three catalytic sites,including Glu71,Asp84 and Asp101.Quantitative qRT-PCR results confirmed that NaLyg gene mRNA was visibly increased after V.harveyi infection.The NaLyg protein purified by prokaryotic expression killed some gram-negative bacterial pathogens by inducing cell wall destruction,including V.harveyi,Aeromonas hydrophila and Edwardsiella tarda.Moreover,the NaLyg protein killed two gram-positive bacteria,Bacillus subtilis and Staphylococcus aureus.Taken together,the experimental results suggested that the NaLyg protein of N.albiflora played an important role in fighting bacterial infections.

Development of SNP parentage assignment techniques in the yellowfin seabream Acanthopagrus latus

Acanthopagrus latus is an essential aquaculture species on the south coast of China.However,there is a lack of systematic breeding of A.latus,which considerably limits the sustainable development of A.latus.As a result,genetic improvements are urgently needed to breed new strains of A.latus with rapid growth and strong resistance to disease.During selective breeding,it is necessary to estimate the genetic parameters of the target trait,which in turn depends on an accurate disentangled pedigree for the selective population.Therefore,it is necessary to establish the parentage assignment technique for A.latus.In this study,95 individuals selected from their parents and their 14 families were used as experimental material.SNPs were developed by genome resequencing,and highly polymorphic SNPs were screened on the basis of optimized filtering parameters.A total of 14392738 SNPs were discovered and 205 SNPs were selected for parentage assignment using the CERVUS software.In the model where the gender of the parents is known,the assignment success rate is 98.61%for the male parent,97.22%for the female parent,and 95.83%for the parent pair.In the model where the gender of the parents is unknown,the assignment success rate is 100%for a single parent and 90.28%for the parent pair.The results of this study were expected to serve as a reference for the breeding of new varieties of A.latus.

Composition of fish species in the Bering and Chukchi Seas and their responses to changes in the ecological environment

Based on trawl surveys in the Bering Sea and Chukchi Sea during the 2010 Chinese National Arctic Research Expedition, fish biodiversity characteristics, such as fish composition, dominant species, biodiversity, and faunal characteristics were conducted. We also discussed the responses of fishes to the quick changes in Arctic climate. The results showed that a total of 41 species in 14 families were recorded in these waters. The dominant species were Hippoglossoides robustus, Boreogadus saida, Myoxocephalus scorpius, Lumpenus fabricii, and Artediellus scaber. There were 35 coldwater species, accounting for 85.37%, and six cold temperate species, occupying 14.63%. The habitat types of fish could be grouped as follows： 35 species of demersal fishes, five benthopelagic fishes, and one pelagic fish. The Shannon-Wiener diversity index （H＇） （range between 0 and 2.18, 1.21 on average） was not high, and descended from south to north. Climate change has caused some fishes to shift along their latitudinal and longitudinal distribution around the Arctic and Subarctic areas, and this could lead to the decline of Arctic fishery resources.

Adhesion mechanisms of Vibrio fluvialis to skin mucus of Epinephelus awoara

Vibriofluvialis incubated in trypticase soy broth （TSB） showed stronger adhesion to the skin mucus of Epinephelus awoara than V. fluvialis grown on trypticase soy agar （TSA）, and this bacterial adhesion was assessed in terms of saturation kinetics. Treating bacteria with antibody against O-antigens resulted in significantly reduced bacterial adhesion. In the early growth stage, the adhering bacteria numbers increased with incubation time, peaked at 24 h, and then dropped sharply. Prior heat treatment of the mucus at various temperatures resulted in different effects on bacterial adhesion. No remarkable effect on bacterial adhesion was detected after treating mucus with protease K and trypsin, whereas periodic acid treatment significantly enhanced bacterial adhesion. Extracts of K fluvialis surface structures at high concentrations significantly inhibited bacterial adhesion, while relatively low concentrations enhanced adhesion. V. fluvialis had strong affinity for a high molecular weight mucus component. These results indicate that V.fluvialis adhered strongly to E. awoara mucus by specific adhesion and that this adhesion was influenced by many factors, including bacterial conditions, mucus, and the environment.

Effects of Temperature on Haemocyte and Granulocyte Counts and Expressions of Immunity-Related Genes in Haemocytes of Scallop Chlamys farreri After Vibrio anguillarum Infection

Bivalve live in aquatic environment and the water temperature can affect their immunity directly.In this research,the scallop Chlamys farreri was injected with 10^4 or 10^7 CFU mL^-1 Vibrio anguillarum and cultured at 11℃,17℃,23℃,and 28℃,respectively.For the control scallop,only phosphate-buffered saline(PBS)was injected.Then total haemocytes and granulocytes were measured by ELISA using monoclonal antibodies.In the meantime,expressions of six immunity-related genes,including lipopolysaccharide andβ-1,3-glucan binding protein(CfLGBP),C-type lectin(CfLec-2),Toll-like receptor(Cf TLR),Lysozyme(CfLYZ),superoxide dismutase(SOD),and phenoloxidase(CfPO)in haemocytes were measured using quantitative real-time PCR.The results showed that total haemocytes counts in 10^4 CFU mL^-1 injection groups showed no differences compared to the control group at all temperatures.However,they varied significantly in 10^7 CFU mL^-1 injection groups at 3 h at 11℃,6–12 h at 17℃,3–48 h at 23℃,and 12–48 h at 28℃.Granulocytes counts in 10^4 CFU mL^-1 injection groups showed no variance compared to the control group at all temperatures,except for 12 h at 23℃,and 24–36 h at 28℃.They were significantly decreased in 10^7 CFU mL^-1 injection groups during 6–48 h at 11℃,12–48 h at 17℃,3–48 h at 23℃,and 3–72 h at 28℃.The expression levels of six immunity-related genes in haemocytes of 10^7 CFU mL^-1 injection groups were significantly higher than those of control group and 10^4 CFU mL^-1 injection groups at all temperatures.The results indicated that infected with high concentration of vibrios,haemocyte counts,granulocyte counts and the expressions of immunity-related genes in scallop C.farreri were significantly affected by environmental temperature.

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly increased the number of erythrocytes, leucocytes, lymphocytes in peripheral blood as well as peripheral serum antibacterial activity and antibody titer of large yellow croaker, and significantly reduced the number of peripheral blood granulocytes as compared with those in the control group. No significant difference in acid phosphytase and superoxide dismutase activity of serum was detected between the two groups. It is suggested that non-specific immune factors including leucocytes and anti-bacteria substance in peripheral blood played important role at the initial stage of infection, and specific immune factors such as antibody then played important role in response to anti-infection at the latter stage.

A Comparative Study of Fish Community in Four Main Estuaries of China Southeastern Coastal Areas and Their Adjacent Waters

Based on literature and survery data of fish composition collected by bottom trawl investigation from 2006 to 2007 infour main estuaries of China southeastern coastal areas and their adjacent waters,changes of the taxonomic diversity across spatialand temporal scales of fish community were analyzed by taxonomic diversity indices.The results are as follows:a total number of1397 fish species(including some freshwater species),belonging to 2 classes,42 orders,186 families and 593 genera,were collected inthe studied sea areas.The species richness increased with lower latitudes,particularly so with Perciformes.There were 339 fish spe-cies in the Yangtze River Estuary and adjacent waters,belonging to 2 classes,31 orders,101 families and 231 genera.There were 535fish species in the Minjiang River Estuary and adjacent waters,belonging to 2 classes,33 orders,133 families and 323 genera.A totalnumber of 803 fish species were collected in the Jiulongjiang River Estuary and adjacent waters,which belonged to 2 classes,35 orders,155 families and 419 genera.And 1021 fish species which belonged to 2 classes,32 orders,153 families,and 466 genera were collectedin the Pearl River Estuary and adjacent waters.The numbers of orders and families of fish species from the northern to the southern wa-ters first increased and then decreased.The average variation in taxonomic distinctness(Λ+)gradually decreased with lower latitudes inthe four estuaries and adjacent waters.There were no significant differences in the average taxonomic distinctness(△+)among the fourestuaries and adjacent waters,and fish fauna were closely related with each other,and all of them belonged to the same zoogeographicalfauna(Indian-Malaysia fauna).Fish composition in the Minjiang River Estuary and adjacent waters was more similar to that in the Jiu-longjiang River Estuary and adjacent waters among the four estuaries.Compared with the historical data,the average taxonomic dis-tinctness of fish community showed a great decrease in the Minjiang River Estuary and the Jiulongjiang River Estuary and their adjacentwaters.

A Sensitive Reversed-Phase High-Performance Liquid Chromatography Method for the Quantitative Determination of Milk Xanthine Oxidase Activity

A new reversed-phase high performance liquid chromatography method was developed to quantitate the activity of xanthine oxidase involved in milk fat globule membrane with xanthine as the substrate and the separation of product (uric acid). The increment of uric acid in the reaction system was used to calculate the total activity of XO. The optimized assay conditions, linearity of detection, recovery of uric acid and chromatogram were developed in text, indicating this method is simple, rapid and efficient. It is an alternative potential method for the determination of the activity of XO in milk.

Composition and distribution of fish species collectedduring the fourth Chinese National Arctic Research Ex-pedition in 2010

There are awareness and concerns caused by the decreasing sea ice coverage around the Arctic and Antarctic due to effects of climate change. Emphasis in this study was on rapid changes in Arctic sea ice coverage and its impacts on the marine ecology during the fourth Chinese National Arctic Research Expedition in 2010. Our purpose was to establish a baseline of Arctic fish compositions, and consequent effects of climate change on the fish community and biogeography. Fish specimens were col- lected using a multinet middle-water trawl, French-type beam trawl, otter trawl, and triangular bottom trawl. In total, 36 tows were carried out along the shelf of the Bering Sea, Bering Strait, and Chukchi Sea in the Arctic Ocean. in total, 41 fish species belonging to 14 families in 7 orders were collected during the expedition. Among them, the Scorpaeniformes, including 17 species, accounted for almost one third of the total number （34.8%）, followed by 14 species of the Perciformes （27.0%）, 5 species of the Pleuronecti- formes （22.3%）, and 2 species of the Gadiformes （15.4%）. The top 6 most abundant species were Hippoglossoides robustus, Bore- gadus saida, Myoxocephalus scorpius, Lumpenus fabricii, Artediellus scaber, and Gymnoeanthus trieuspis. Abundant species var- ied according to the different fishing methods; numbers of families and species recorded did not differ with the various fishing methods; species and abundances decreased with depth and latitude; and species extending over their known geographic ranges were observed during the expedition. Station information, species list, and color photographs of all fishes are provided.

Spacial Distribution Characteristics, Activity and Degradation Capability of the Biofilm in Integrated Vertical-flow Constructed Wetland

The spacial distribution characteristics, activity and degradation capability of the biofilm in integrated vertical-flow constructed wetland were investigated. Results showed that the biofilm widely distributed in the substrate of integrated vertical-flow constructed wetland and mainly in the 0-10 cm top layer where the activity ofdehydrogenase of the biofilm was also higher than that of other layers. The water quality could also affect the activity of the biofilm, for the TF（1,3,5-Triphenylformazan） amount reduced by the biofim incubated in water of the higher eutrophication was larger, too. The PCP （pentachlorophenol） removal rate by the substrate with biofilm was 1.5 times that without biofilm. In total, the biofilm of the down-flow chamber appeared larger biomass, higher dehydrogenase activity and stronger degradation capability of organic contamination than that in up-flow chamber and it was the major place for removal of the organic matters in waste water.

Antioxidant effects of the aqueous extract of turmeric against hydrogen peroxide-induced oxidative stress in spotted seabass(Lateolabrax maculatus)

Intensive aquaculture-induced oxidative stress is detrimental to fish health and yield.Medicinal plants show promise as natural health boosters and antioxidants in the aquaculture industry.Therefore,this work investigated the effects of turmeric aqueous extract(TAE)on the growth performance,antioxidant status,and hydrogen peroxide(H2O2)-induced oxidative stress in spotted seabass(Lateolabrax maculatus).Fish were fed diets supplemented with 0(Con),2(TAE2),or 4(TAE4)g/kg TAE for eight weeks,then were injected with H2O2.The results showed that dietary supplementation of TAE did not affect fish growth,feed utilization,or body composition.TAE treatment increased liver antioxidant enzyme activities and decreased liver malondialdehyde content and serum levels of glutamate oxalate transaminase,glutamate pyruvate transaminase,and lactate dehydrogenase.Furthermore,the increases in mortality,liver malondialdehyde content,and serum biomarkers of liver injury in the H2O2-treated fish were inhibited as a consequence of the TAE treatment.In addition,TAE treatment activated the Nrf2/Keap1 pathway in the liver,supported by the up-regulated expression of nrf2,ho-1,and gclc,and down-regulated keap1 expression.Overall,dietary incorporation of TAE protected against H2O2-induced oxidative stress in spotted seabass probably by enhancing antioxidant capacity through the Nrf2/Keap1 pathway.

Growth performance,fatty acid composition,and lipid metabolism are altered in groupers(Epinephelus coioides)by dietary fish oil replacement with palm oil

In this study,we conducted a 56-d feeding trial to investigate the effects of replacing the fish oil(FO)with palm oil(PO)on the performance,tissue fatty acid(FA)composition,and mRNA levels of genes related to hepatic lipid metabolism in grouper(Epinephelus coioides).Five isolipidic(13%crude lipid)and isonitrogenous(48%CP)diets were formulated by incrementally adding PO to the control diet(25%fish meal and 9%added FO)to replace FO in the control diets.Triplicate groups of 30 groupers(initial weight:12.6±0.1 g)were fed one of the diets twice daily,to apparent satiety.The replacement of FO with 50%PO revealed maximum growth without affecting the performance and whole-body proximate compositions,and replacing FO with 100%PO revealed a comparable(P>0.05)growth with that of the control diet,suggesting PO as a suitable alternative to FO.The analysis of FA profiles in the dorsal muscle and liver though reflected the FA profile of the diet,PO substitutions above 50%could compromise(P<0.05)the FA profile in the liver and flesh of the fish species in comparison with the control diet.Furthermore,the mRNA levels of FAS,G6PD,LPL,PPARА,and D6FAD genes in the liver had positive linear and/or quadratic responses,but the SCD,HSL,ATGL,FABP,SREBP-1C and ELOVL5 had the opposite trend,with increasing dietary PO inclusion levels,whereas the mRNA level of ACC was not affected by dietary treatments.The optimal level of PO substitution for FO was estimated to be 47.1%of the feed,based on the regression analysis of percent weight gains against dietary PO inclusion levels;however,it might affect the FA profile in the liver and flesh of the fish species,and further study is required to investigate whether the changes in tissue FA composition will affect the welfare and market value over a production cycle of grouper.