Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.     

A Successful Microbound Diet for the Larval Culture of Chinese Shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselrnis chuii, rotifer （Brachionus plicatilis） and Arternia sp. Larvae of 0 to 13d post-hatch （dph） were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L^-1 in tanks, each containing 50 L sterilized seawater with salinity 30-32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13 dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13dph, which was not significantly different from that of larvae fed on live foods （63.55%）. The body length and development index （DI） of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13 dph （P〈 0.05）. The mean dry weight loss of the microbound diet was 9.2% after 90 min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 52.23% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

Screening White Spot Syndrome Virus(WSSV)-Resistant Molecular Markers from Fenneropenaeus chinensis

White spot syndrome virus(WSSV)-resistant molecular markers were screened from the selectively bred new variety ‘Huanghai No. 2' of Fenneropenaeus chinensis using unlabeled-probe high-resolution melting(HRM) technique. After the artificial infection with WSSV, the first 96 dead shrimps and the last 96 surviving shrimps were collected, representing WSSV-susceptible and-resistant populations, respectively. The genotypes at well-developed 39 single nucleotide polymorphisms(SNPs) loci were obtained. As revealed in the Chi-square test, 3 SNPs, genotype A/A of contig C364-89 AT, genotype A/A of C2635-527 CA and genotype C/T of contig C12355-592 CT, were positively correlated with disease-resistance traits. Other 2 SNPs, genotype G/G of contig C283-145 AG and genotype C/C of contig C12355-592 CT, were negatively correlated. Moreover, analysis with Blast X program for disease-resistant SNPs indicated that 3 contigs, Contig283, Contig364 and Contig12355, matched to the functional genes of effector caspase of Penaeus monodon, peptide transporter family 1-like protein, and 40 S ribosomal protein S2 of Perca flavescens with high sequence similarity. The results will be helpful to provide theoretical and technical supports for molecular marker-assisted selective breeding of F. chinensis.     

Partial Sequence Analysis of Mitochondrial COI Gene of the Chinese Shrimp, Fenneropenaeus Chinensis

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235bp (28.0% ),.307bp (36.6%), 138bp (16.5%) and 158bp (18.9%), respectively. Furthermore, 556bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

Peptidomic analysis of Chinese shrimp (Fenneropenaeus chinensis) hemolymph by magnetic bead-based MALDI-TOF MS

Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based purification(ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) to characterize shrimp hemolymph peptides.Shrimp serum and plasma were used as the source of samples for comparative analysis,and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis.To screen potential specific biomarkers in serum of immune-challenged shrimps,we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps.The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software.Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide(LPS)-infection.The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%.Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph,and will help to enable a better understanding of the innate immune response of shrimps.     

Cloning,characterization,and expression analysis of the DEAD-box family genes,Fc-vasa and Fc-PL10a,in Chinese shrimp (Fenneropenaeus chinensis)

RNA helicases of the DEAD-box and related families are involved in various cellular processes including DNA replication,DNA repair,and RNA processing.However,the function of DEAD-box proteins in aquaculture species is poorly understood at molecular level.We obtained the full-length cDNA sequences of two genes encoding helicase-related proteins,Fc-vasa and Fc-PL10a,from the testes of Chinese shrimp,Fenneropenaeus chinensis.The two predicted amino acid sequences contain all the conserved motifs characterized ...     

Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)

The clotting protein (CP) plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis (named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca²⁺, suggesting that the clotting reaction is catalyzed by a Ca²⁺-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers. The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.     

Effects of Salinity Fluctuation Frequency on the Osmolarity, Na^＋-K^＋ -ATPase Activity and HSP70 Expression in Juvenile Chinese Shrimp, Fenneropenaeus chinensis

Experiments were conducted to examine the effects of salinity fluctuation frequency on the osmolarity, Na+-K+-ATPase activity and HSP70 of Chinese shrimp Fenneropenaeus chinensis with initial wet body weight of 1.460 g ± 0.091 g. The salinity in the control group (D0) was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8 d, respectively. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8...     

Isolation, cloning and sequencing of AFLP markers related to disease-resistance traits in Fenneropenaeus chinensis

Amplified fragment length polymorphisms （AFLP） technique was used to analyze the fingerprinting of four successive generations of Fenneropenaeus chinensis to reveal their disease-resistance traits. Some loci showed quite different genetic frequencies due to artificial selection, which implied that these fragments were putative markers related to the disease-resistance trait. We developed a simple and effective method to further characterize these AFLP fragments. Specific AFLP bands were cut directly from polyacrylamide gels, re-amplified, cloned and sequenced. Eight putative genetic markers were sequenced and their sizes ranged from 63 to 209 bp. The sequences were submitted to dbGSS （database of Genome Sequence Survey）; and the BLAST analysis showed low similarity to the function genes, indicating these markers were tightly linked to a disease-resistance trait but were not functional genes.     

Nitrogen and Phosphorus Budget of a Polyculture System of Sea Cucumber （Apostichopus japonicus）, Jellyfish （Rhopilema esculenta） and Shrimp （Fenneropenaeus chinensis）

The nitrogen(N) and phosphorus(P) budget and the ecological efficiency of a polyculture system of sea cucumber(Apostichopus japonicus), jellyfish(Rhopilema esculenta) and shrimp(Fenneropenaeus chinensis) were studied in a cofferdam, 120.2 ha in size. The nutrients were supplied by spring tide inflow. In total, 139600 kg N yr-1 and 9730 kg P yr-1 input to the system; while 118900 kg N yr-1 and 2840 kg P yr-1 outflowed from the system concurrently, thus the outflow was 85.7%(N) and 29.2%(P) of inflow. The production of N and P was 889.5 kg yr-1 and 49.28 kg yr-1(sea cucumber) and 204 kg yr-1 and 18.03 kg yr-1(jellyfish and shrimp), respectively. The utilization rate of N and P by polycultured animals was 7.8‰ and 6.9‰, respectively, 21.9% and 38% higher than that of monocultured sea cucumber. Our results indicated that the polyculture system was an efficient culture system of animals and a remediation system of coastal environment as well; it scavenged 14.3% and 70.8% of N and P, respectively. Such an ecological efficiency may be improved further by increasing either the stocking density or the size of sea cucumber or both.     

Effects of Salinity Fluctuation Frequency on the Growth, Molting Rate and Hemolymph 20-Hydroxyecdysone Concentration in Juvenile Chinese shrimp, Fenneropenaeus chinensis

This experiment was conducted to examine the effects of salinity fluctuation frequency on the growth, molting rate and hemolymph 20-hydroxyecdysone (20-HE) concentration for juvenile Fenneropenaeus chinensis with an initial wet body weight of 1.115 g±0.012 g. The salinity of the control treatment (represented by D0) was 28 throughout the experiment, whereas treatments D2,D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8d, respectively. The range of salinity fluctuation was 4. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8d, respectively; each original salinity was decreased abruptly to salinity 24, which first lasted for another 2d and was then raised to its initial value 28. This constituted a salinity fluctuation cycle and afterwards the cycle repeated. It was found that: 1) The weight gain of shrimp in treatment D4 was 202.9% and significantly higher than that in treatment D0 (P<0.05). The molting rate in treatment D2 was the lowest, while that in treatment D6 was the highest and significantly higher than those in treatments D0 and D2 (P<0.05). 2) The hemolymph 20-HE concentration of shrimps in all treatments was at low levels and increased first gradually during the inter-molt period and then increased sharply. It reached a peak value during the pre-molt stage, and then decreased abruptly during the post-molt stage when the lowest level occurred. 3) During the post-molt stage, the hemolymph 20-HE concentration in treatment D2 was 7.47pgμL-1 and significantly higher than that in treatment D0(P<0.05). There were, however, no significant differences in hemolymph 20-HE concentration among all five treatments during the inter-molt stage (P>0.05). During the pre-molt stage, the hemolymph 20-HE concentration in treatment D6 was significantly higher than that in treatment D0(P<0.05), whereas the hemolymph 20-HE concentration in treatment D2 was significantly lower than that in other treatments (P<0.05). Adequate salinity fluctuation promoted the molting rate through increasing hemolymph 20-HE concentration at the pre-molt stage.     

Effects of salinity fluctuation amplitudes on growth, osmolarity, Na＋-K＋-ATPase activity and Hsp70 of juvenile Chinese shrimp Fenneropenaeus chinensis Osbeck

The effects of various salinity fluctuation amplitudes (2, 4, 6 and 8) on the growth, osmolarity, Na⁺-K⁺-ATPase activity and Hsp70 of juvenile Fenneropenaeus chinensis cultured in seawater with a salinity of 20 were studied. The results show that weight gain in the salinity fluctuation treatments was better than that in control; in particular, the weight gain of treatments S4 and S0, at 231.8% and 196.3%, respectively, was significantly different (P<0.05). The hemolymph osmolarity of treatments S0, S2, S4, S6 and S8 was 635.4, 630.8, 623.6, 614.4 and 600.3 mOsm/kg, respectively, and decreased with increasing salinity fluctuation amplitude. The level of Na⁺-K⁺-ATPase activity in gills of F. chinensis was higher than that in hepatopancreas, but there were no significant differences among all treatments, either in gills or hepatopancreas (P>0.05). The relative level of Hsp70 in treatment S4 was 48.4% and 40.4% higher than control in muscle and eyestalks, respectively, with the highest values being recorded under a salinity fluctuation amplitude of 4.     

Genetic improvement on Chinese shrimp （Fenneropenaeus chinensis）： growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea （WYP）, and the west coast of the Korean Peninsula and coast （WKN）. The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea （FKN）, the Huang Hai （the Yellow Sea in Chinese） No.1 （HH1）, and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length （BL）, carapace length （CL）. carapace width （CW）, height of the second and third abdominal segment （HST）, width of the second and third abdominal segment （WST）, length of the first abdominal segment （LF）, length of the last abdominal segment （LL）, live body weight （BW）, and survival rate were measured, Different combinations were statistically performed with ANOVA and Duncan＇s Multiple Range Test. The results show that the survival rate of JK98（♀）×WKN（♂） was the highest, followed by WYP（♀）×WKN（♂）, FKN（♀）×WYP（♂）, FKN（♀）×HHI（♂） and WYP（♀）×FKN（♂）; the body weight of FKN（♀）×HHI（♂） was the highest, followed by FKN（♀）×WYP（♂）, WYP（♀）×WKN（♂）, WYP（♀）×FKN（♂） and JK98（♀）×WKN（♂）; the total length had the same ranking as the body weight. All growth traits in hybrids JK98（♀）×WKN（♂） were the lowest among all combinations. F1 hybrids had significant difference （P〈0.05） in BL, CL, HST, LL, and BW; and insignificant difference （P〉0.05） in other growth traits and survival rate. The results of Duncan＇s Multiple Range Test are that BL and CL of J     

A STUDY ON PATHOGENS OF CHINESE PRAWN (PENAEUS CHINENSIS) VIRUS DISEASES

This pathogenic study shows that the viral diseases of Chinese prawns (Penaeus chinensis. O'sbeck)is due to three kinds of viruses: epithelium envelope baculovirus of Penaeus chinensis (EEBV-PC, de-tected by the authors in 1993), infections hypodermal and hematopoietic necrosis virus, andhepatopancreatic parvo-like virus, and that the first two viruses seem to be the main pathogens of theepidemic in the northern regions in 1993.     

Movements of the Western Pacific Warm Pool Centroid and Their Relationship to Sea Surface Temperature Changes in Ni(n)o Regions

By using monthly historical sea surface temperature (SST) data for the years from 1950 to 2000, the Western Pacific Warm Pool (WPWP) climatology and anomalies are studied in this paper. The analysis of WPWP centroid (WPWPC) movement anomalies and the Nino-3 region SST anomalies( SSTA) seems to reveal a close, linear relation between the zonal WPWPC and Nino-3 region SSTA, which suggests that a 9' anomaly of the zonal displacement from the climatological position of the WPWPC corresponds to about a 1 ℃ anomaly in the Nino-3 region area-mean SST. This study connects the WPWPC zonal displacement with the Nino-3 SSTA, and it may be helpful in better understanding the fact that the WPWP eastward extension is conducive to the Nino-3 region SST increase during an El Nino-Southern Oscillation (ENSO) event.     

Comparative fatty acid composition of fourSargassum species (Fucales,Phaeophyta)

Fatty acid composition of fourSargassum species from Qingdao and Shidao, Shandong Province was investigated. 16:0 (palmitic acid) was the major saturated fatty acid. C₁₈ and C₂₀ were the main polyunsaturated fatty acids (PUFAs). Arachidonic acid and eicosapentaenoic acid predominated among polyenoic acids in all the algal species examined, except forSargassum sp. which had low concentration of eicosapentaenoic acid. Contribution No. 2354 from the Institute of Oceanology, Chinese Academy of Sciences     

基于Qi(xi,yi)函数的约束Delaunay三角剖分算法

利用Qi算法的性质,提出了一种基于Qi(xi,yi)函数的约束Delaunay三角剖分算法。经过比较分析,本文提出的算法降低了时间复杂度,提高了执行效率。     

Middle and Late Cambrian trilobite communities from Fenghuang (Dama) of western Hunan, China

Middle and Late Cambrian trilobite faunas from the Dama section in western Hunanare are composed of 63 genera and subgenera, 84 species, and two forms of gen. et sp. indetermined. Which include a mixture of cosmopolitan agnostoids and polymeroids. Three new species,Hardyoides damaensis,Meringaspis damaensis and Rhyssometopus (Rodtrifinis) nitidus, are described by present author in 2004. Besides, Erixanium is recognised for the first time in study area, which is significant for biostratigraphical correlation of the Late Cambrian in the Austalo-Asia region. According to distribution regularity vertical, the Middle and Late Cambrian trilobites can be divided into 9 trilobite communities for the Dama section. Based on the trilobite communities analysis, the author concludes that from the east Mt. Lailong to Dama through Fenghuang, further east, reach in Chenxi,the palaeoenvironments were changed from a shallow-water of oxygen-rich (platform margin slope ) into a deep-water of oxygen-poor setting (basin).