Preliminary Study on the Water Drag of Schlegel＇s Black Rockfish Sebastes schlegeli

Drag forces acting on Schlegel‘s black rockfish Sebastes schlegeli were studied. A new drag force transducer was designed and used to measure the water drag on Schlegel‘s black rockfish in a vertical recirculating flume tank. Fourteen individuals were investigated, yielding two mean drag coefficients referred to the cross-sectional area and volume^2/3 respectively at water velocities ranging from 0.3 to 1.0 ms^-1 The drag coefficients can be used for estimating the drag forces acting on Sebastes schlegeli in water.     

Microbiota characteristics in Sebastes schlegelii intestine in early life stages

Journal of Oceanology and Limnology - The structure of intestinal microbiota of black rockfish Sebastes schlegelii in five early development stages were determined in high throughput sequencing...     

A microsatellite genetic linkage map of black rockfish (Sebastes schlegeli)

Ovoviviparous black rockfish (Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers. In this study, a large number of microsatellite markers of black rockfish were isolated by constructing microsatellite-enriched libraries. Female- and male-specific genetic linkage maps were constructed using 435 microsatellite markers genotyped in a full-sib family of the fish species. The female linkage map contained 140 microsatellite markers, in which 23 linkage groups had a total genetic length of 1334.1 cM and average inter-marker space of 13.3 cM. The male linkage map contained 156 microsatellite markers, in which 25 linkage groups had a total genetic length of 1359.6 cM and average inter-marker distance of 12.4 cM. The genome coverage of the female and male linkage maps was 68.6% and 69.3%, respectively. The female-to-male ratio of the recombination rate was approximately 1.07:1 in adjacent microsatellite markers. This paper presents the first genetic linkage map of microsatellites in black rockfish. The collection of polymorphic markers and sex-specific linkage maps of black rockfish could be useful for further investigations on parental assignment, population genetics, quantitative trait loci mapping, and marker-assisted selection in related breeding programs.     

Non-specific immune response of bullfrog <Emphasis Type="Italic">Rana catesbeiana</Emphasis> to intraperitoneal injection of bacterium <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally （i.p.） with 0.2 ml bacterial suspension at a density of 5.2 × 10^6 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution （0.85%, w/v）. Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection （dpi） for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase （SOD） activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.     

Immune response of <Emphasis Type="Italic">Pseudosciaena crocea</Emphasis> to the injection of <Emphasis Type="Italic">Vibrio alginolyticus</Emphasis>

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. Supported by the National High Technology Research and Development Program of China (863 program, No. 2007AA09Z115) and Technology Program of Xiamen (No. 3502Z73019)     

Preliminary Study on the Water Drag of Schlegel's Black Rockfish Sebastes schlegeli

1　Introduction　FishcageculturesystemhasbeenwidelyusedinmaricultureindustryinChina .Indesigningthecagesystem ,thewaterdragactingonit,includingthatonculturedfish ,shouldbetakenintoaccount.Avarietyofmethodshavebeenusedtodeterminethewaterdragofanimalsrangin…     

Detection of lymphocystis disease virus in Japanese flounder Paralichthys olivaceus and other marine teleosts from northern China

We isolated a strain of lymphocystis disease virus (LCDV) from Japanese flounder (Paralichthys olivaceus) cultured in northern China. Based on published sequences of major capsid protein (MCP) gene of LCDV-cn (GenBank: AF126405), we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization (ISH) to detect LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral supernatant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus (OFIV and RSIV), Iridovirus (CzIV and WIV), Ranavirus (TFV and FV3), and Lymphocystivirus (8 LCDV). Based on the alignment, we confirmed the PCR product was from Lymphocystivirus (GenBank accession number DQ279090 (LCDV-HD)). Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen, stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel’s black rockfish (Sebastes schlegeli Higendorf), redwing sea robin (Lepidotrigla microptera Günther) and turbot (Scophthalmus maximus) using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method.     

Photosynthetic and metabolic responses of eelgrass Zostera marina L. to short-term high-temperature exposure

In recent years, extreme heat events have occurred worldwide and the ocean temperature has been rising, causing stress on the photosynthesis and growth of seagrass. Metabolomics enables detection of metabolic changes under environmental stress. In this study, the photosynthetic physiology and metabolic changes of the eelgrass Zostera marina L. in response to 48 h exposure to 32°C were investigated. The results showed that high temperature induced signi?cant inhibition of photosynthetic effciency(Δ F/F′ m)(23.9%lower than the control), enhanced respiration(58.3% higher), and decreased carbohydrate decomposition products and tricarboxylic acid(TCA) cycle intermediate products, indicating that the energy supply of the eelgrass may be insuffcient at high temperature. In addition, high temperature decreased stearic acid and linoleic acid in eelgrass, suggesting the composition of the membrane system of eelgrass may change at high temperature and implying that high temperature may cause the membrane system to be unstable.     

Triterpene-Enriched Olive Extract as an Immunopotentiator in Black Sea Bream ( Acanthopagrus schlegelii)

Recently,the use of natural immunopotentiators,such as plant extracts,is predicted to offer high disease-prevention potential in aquaculture.However,few reports on the immunomodulatory impacts of olive extract(OE)on teleost are available.Therefore,the effects of dietary intake of triterpene-enriched OE on black sea bream(Acanthopagrus schlegelii)were investigated in this study.Our data showed that total blood cell counts,in vivo content of lysozyme,activities of antiprotease and myeloperoxidase,and contents of IL-4 and IL-6 were significantly up-regulated by dietary intake of triterpene-enriched OE.Additionally,the hampered immune response induced by cadmium exposure was significantly mitigated by the administration of OE as were indicated by partially or completely rescued immune-relating parameters.Furthermore,the expressions of immune-related genes encoding NF-κB inhibitor alpha(IkBα),tumor necrosis factorα(TNFα),cyclooxygenase-2(COX-2),and proto-oncogene protein c-fos(FOS)were found to be significantly up-regulated by the dietary intake of OE.In general,the results suggested that the dietary intake of triterpene-enriched OE has immune enhancing effect in black sea bream.Such effect may be realized by 1)increasing the total counts of diverse blood cells;2)activating nonspecific immune biomolecules;and 3)affecting signaling pathways such as IKK and ERK and subsequently inducing IL-4 and IL-6 biosyntheses.     

Genes of the adaptive immune system are expressed early in zebrafish larval development following lipopolysaccharide stimulation

Information regarding immunocompetence of the adaptive immune system(AIS) in zebrafish Danio rerio remains limited.Here,we stimulated an immune response in fish embryos,larvae and adults using lipopolysaccharide(LPS) and measured the upregulation of a number of AIS-related genes(Rag2,AID,TCRAC,IgLC-1,mIg,sIg,IgZ and DAB) 3 and 18 h later.We found that all of the genes evaluated were strongly induced following LPS stimulation,with most of them responding at 8 d post fertilization.This confirms that a functional adaptive immune response is present in D.rerio larvae,and provides a window for further functional analyses.     

Gene cloning and prokaryotic expression of recombinant flagellin A from Vibrio parahaemolyticus

The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction of the host immune response. Thus, flagellin proteins are an ideal target for vaccines. We amplified the complete flagellin subunit gene (flaA) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 62.78 kDa. We purified and characterized the protein using Ni-NTA affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for further studies into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus. In addition, the purified FlaA protein can be used for further functional and structural studies.     

Characterization of Pythium chondricola associated with red rot disease of Pyropia yezoensis (Ueda) (Bangiales,Rhodophyta) from Lianyungang,China

Pyropia yezoensis(formerly Porphyra yezoensis) is an economically important red alga that is cultured extensively in China. The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses. An incidence of red rot disease was found in a P. yezoensis farm from mid-November to mid-December 2015 at Lianyungang, Jiangsu Province, China. Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen, leading to red rot symptoms. The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox1). In artificial infection experiments on the P. yezoensis blades, the P. chondricola isolate was able to cause the same characteristic histopathology seen in natural infections. P. chondricola grew well at a wide range of temperatures in the range 8-31℃, salinities at 0-45 and pH 5-9. In an orthogonal test used to determine the effects of environmental factors(temperature, salinity, and zoospore concentration)on infection, the data revealed that temperature was the most important factor to affect red rot disease development, with the optimal conditions for disease expansion being 20℃, 35 salinity, and a zoospore concentration of 10~6 zoospores/mL. The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia, which will provide support to maintain the healthy development of the Pyropia industry in China.     

Identification and sequence of a cDNA clone corresponding to a gene involved in development ofUndaria pinnatifida

During the induction of gamete-producing gametangia, induced gametophytes were collected at 4 days intervals (0,4,8,12 d) and total RNAs were isolated by CsCl gradient ultracentrifugation. Some stage-specific expressed mRNAs were identified by differential display of mRNAs from different developing stages of the gametophytes. The cDNA of one specific mRNA was verified, cloned and sequenced. This gene was specifically expressed during 4 days of induction, and had partial homologous sequence with tobacco IAA-binding protein gene. It suggests that this cDNA may represent a gene which is related to the IAA regulating function during the development of the gametophytes.

     

Effects of dietary β-glucan and glycyrrhizin on non-specific immunity and disease resistance of the sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) challenged with <Emphasis Type="Italic">Vibrio splendidus</Emphasis>

Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×10⁸ cfu / sea cucumber for 15 days. Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), lysozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0^th, 5^th, 10^th and 15^th days after injection. Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet. On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V. splendidus decreased in value significantly. On the 15^th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0^th day. For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial. After injecting with V. splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V. splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin. There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.     

Effects of hypoxia on dopamine concentration and the immune response of White Shrimp (Litopenaeus vannamei)

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolymph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5 mg L⁻¹ DO groups at 12 h and 6 h, and then returned to normal after 24 h and 12 h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0 mg L⁻¹ DO group became stable after 12 h, while granular cells did so after 24 h. The THC and different haemocyte count (DHC) in the 1.5 mg L⁻¹ DO group became stable after 24 h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5 mg L⁻¹ DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 h, respectively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activity were obviously related to the exposure time under hypoxic conditions.