Three Rana grylio virus (RGV) isolates and lymphocystis disease virus (LCDV-C) were molecularly characterized by antigenicity comparison, Western blot detection of viral polypeptides, restriction fragment length polymorphism analysis of viral genomes, and MCP sequence analysis. Significant antigenicity differences existed among the three RGV isolates and LCDV-C. Western blot detection indicated that the viral polypeptides of three RGV isolates could be recognized by the anti-RGV9807 serum, whereas no bands were observed in the LCDV-C, and significant differences exist among the band patterns of three RGV isolates. Restriction fragment length polymorphism (RFLP) analysis was performed by digesting genomic DNA of the four iridovirus isolates with restriction endonucleases HindIII, KpnI, XbaI and BamHI. On the whole, obvious discrepancies existed between LCDV-C and RGV isolates, and some significant band pattern differences were also revealed between RGV9808 and RGV9506 (or RGV9807) in the profiles of restriction endonucleases Xbal, Kpn I and BamHI. PCR amplification and sequence analysis of MCP gene sequence further revealed their phylogenetic relationship among the three RGV isolates, LCDV-C and other iridoviruses. RGV9506, RGV9807 and RGV9808 are clustered together with other ranaviruses, such as FV3, BIV, TFV and ENHV, although the RGV9808 is more close to EHNV than to other ranaviruses. Additionally, LCDV-C is clustered with LCDV-1, the type species of genus Lymphocystisvirus. The current study provides clear evidence that significant genetic difference exists among the three RGV isolates. Therefore, further work on comparative genomic studies will contribute significantly to understanding of their taxonomic position and pathological mechanism. (C) 2005 Elsevier B.V. All rights reserved.
A growth trial was conducted to estimate the optimum requirement of dietary available phosphorus (P) for black seabream (Sparus macrocephalus) in indoor net cages (1.5x1.0x1.0 m). Triplicate groups of black seabream (11.45 +/- 0.02 g) were fed diets containing graded levels (0.18, 0.36, 0.54, 0.72, 0.89 and 1.07%) of available P to satiation for 8 weeks. The basal diet (diet 1), containing 0.18% available P, was supplemented with graded levels of monosodium phosphate (NaH2PO4 2H(2)O) to formulate five experimental diets. The fish were fed twice daily (08:00 h and 16:00 h) and reared in seawater (salinity, 26-29 g l(-1)) at a temperature of 28 +/- 1 degrees C. Dissolved oxygen during the experiment was above 5 mg l(-1). The specific growth rate (SGR), weight gain (WG), feed efficiency (FE) and protein efficiency ratio (PER) were all significantly improved by dietary phosphorus up to 0.54% (P<0.05) and then leveled off beyond this level. Hepatosomatic index (HSI) was inversely correlated with dietary phosphorus levels (P< 0.05). Efficiency of P utilization stabled in fish fed diets containing 0.18%-0.54% available P and then decreased dramatically with further supplementation of dietary phosphorus. Body composition analysis showed that the whole-body lipid, ash, calcium and phosphorus contents were all significantly affected by dietary available P concentration (P0.05). Dietary phosphorus levels also affected the mineralization of vertebrae, skin and scale (P<0.05). Ca/P ratios in vertebrae and scale were not influenced by dietary P supplementation, while skin Ca/P ratio increased statistically with dietary available P levels (quadratic effect, P<0.001). The blood chemistry analysis showed that dietary available P had distinct effects on enzyme activities of alkaline phosphatase (ALP) and plasma lysozyme (LSZ), as well as contents of triacyglycerol (TG) and total cholesterol (T-CHO) (P<0.05). Broken-line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 0.55%. Quadratic analysis based on P contents in whole fish, vertebrae or scale indicated that the requirements were 0.81, 0.87 and 0.88%, respectively. Signs of phosphorus deficiency were characterized by poor growth, slightly reduced mineralization and an increase in body lipid content. (C) 2008 Published by Elsevier B.V.
Streptococcosis became an increasingly significant health problem in intensive aquaculture in China. Fifteen strains of Gram-positive, chain-forming coccus were isolated from moribund Amur sturgeon, Acipenser schrenckii, fanned with high density in central China. The coccoid microorganism was identified as Streptococcus dysgalactiae by means of physiological. biochemical properties and molecular analysis; furthermore, this coccus was confirmed as pathogen of sturgeon by challenge experiments and its infection potential on the cyprinid was also evaluated. To our knowledge, this was the first report of S. dysgalactiae linked to diseased A. schrenckii. (C) 2009 Elsevier B.V. All rights reserved.
A growth trial lasting for 12 weeks was conducted in a semi flow-through system to determine the dietary potassium requirement of juvenile grass carp (Ctenopharyngodon idellus) with an initial body weight of 4.8 g. Seven isonitrogenous and isoenergetic semi-purified diets were compounded with different dietary potassium levels: 1.50, 2.32, 3.28, 5.30, 7.12, 9.45 and 10.44 g kg(-1) diet. The results showed that specific growth rate and feed efficiency of grass carp were significantly (P 0.05). The Na+-K+ ATPase activity value in the gills varied significantly (P < 0.05) with dietary potassium level and showed an increasing and then decreasing trend, and the highest value was in the 7.12 g kg(-1) group. Dietary potassium level showed significant (P < 0.05) effect on serum SOD, ALP and AST activities. SOD and ALP activities increased and then decreased as potassium level increased, while AST decreased first and then increased. Serum TP and TC contents increased and then decreased with potassium level (P 0.05). Body potassium and moisture contents increased and lipid content decreased with dietary potassium content, while ash content in fish body increased first and then decreased as potassium content increased (P < 0.05). Body protein in the 10.44 g kg(-1) group was significantly lowered (P < 0.05). However, calcium, magnesium, phosphorus and sodium contents in dry matter were not affected by dietary potassium level. When the potassium content of water was 1.86-3.10 mg L-1, judging from the polynomial regression analysis for the specific growth rate of fish and the Na+-K+ ATPase activity in the gill, and the broken-line analysis for body potassium content, the optimal dietary potassium content for juvenile grass carp was 4.65, 7.27 and 5.98 g kg(-1) diet respectively. (C) 2014 Elsevier B.V. All rights reserved.
An 8-week experiment was carried out to investigate the effects of light intensity on growth, survival and skin color of Chinese longsnout catfish juveniles. Five light intensities, 0.15, 0.98, 2.46, 3.82 and 5.28 mu mol.s(-1).m(-2) (5, 74, 198, 312 and 434 lx, respectively), were tested in triplicates. Fish (4.8 +/- 0.01 g) were fed to satiation twice a day (0900, 1600 h). The photoperiod was 12L:12D (0800-2000 h). At the end of the experiment, three fish per tank were sampled to measure skin color by instrumental color analysis. The results showed that growth rate was significantly reduced at lower or higher intensities while light intensity did not affect the survival. The skin color of Chinese longsnout catfish was darkest under 434 lx. It is concluded that light intensity significantly affected growth and optimal light intensity for Chinese longsnout catfish juveniles was about 312 lx. (C) 2005 Elsevier B.V All rights reserved.
The chromosomes of the artificial triploid carp consist of three complete genomes from Xingguo red carp (Cyprinus carpio), mirror carp (Cyprinus carpio) and red crucian carp (Carassius auratus). Starch and polyacrylamide gel analysis reveals that some SOD and s-MDH alleles of red crucian carp are undetectable in tissues of the artificial multiple triploid carp. It was found that the majority of oocytes of female artificial multiple triploid carp (XXX) developed to maturity, but spermatocytes of the male (XXY) developed abnormally and no spermatozoa were found. The external features of the triploid carp egg are quite different from those of normal diploid eggs. Sections of fertilized eggs reveal that the majority of eggs are polyspermous. Irradiated sperm was used to induce the gynogenesis of the artificial multiple triploid carp eggs. The eggs cleaved and developed normally, but the numbers of chromosomes of gastrular cells were variable, i.e., 50, 75, 100 and 150. This suggests that the behavior of the chromosomes in meiosis is polytypic, and some female nuclei of the eggs remain triploid after activation. All viable offspring of gynogenetic artificial multiple triploid carp are triploid and no segregation was found in their appearances, scale-covering type and body coloration.
A goose-type lysozyme (g-lysozyme) gene has been cloned from the mandarin fish (Siniperca chuatsi), with its recombinant protein expressed in Escherichia coli. From the first transcription initiation site, the mandarin fish g-lysozyme gene extends 1307 nucleotides to the end of the 3' untranslated region, and it contains 5 exons and 4 introns. The open reading frame of the glysozyme transcript has 582 nucleotides which encode a 194 amino acid peptide. The 5' flanking region of mandarin fish glysozyme gene shows several common transcriptional factor binding sites when compared with that from Japanese flounder (Paralichthys olivaceus). The recombinant mandarin fish g-lysozyme was expressed in E. coli by using pET-32a vector, and the purified recombinant g-lysozyme shows lytic activity against Micrococcus lysodeikticus. (c) 2005 Elsevier B.V All rights reserved.
The objective of this study was to develop type I markers for genome mapping and other genetic studies of Penaeus monodon. Primers were designed based on expressed sequence tags (ESTs) from a P monodon cephalothorax cDNA library to amplify 100-300 bp products. 34 of the primer pairs successfully amplified PCR products from genomic DNA. Single-strand conformation polymorphism analysis showed that similar to 30% of the ESTs tested exhibit polymorphism in a test panel of P monodon individuals. Mendelian inheritance of the EST-derived markers has been established in two international reference mapping families of P monodon, and mapping of these markers is in progress. Some ESTs were successfully amplified from other Penaeus species (P. chinensis, P japonicus and P vannamei), indicating that the markers are applicable in cross-species comparison. Two populations of P. japonicus could be differentiated using one of the ESTS. In conclusion, the polymorphic EST markers developed in this study are applicable in genome mapping and population genetic studies of penaeid shrimp. (C) 2002 Elsevier Science B.V. All rights reserved.