Effects of three methionine sources in diets on temporal B0AT1 and ASCT2 mRNA expression in different intestinal segments of cobia,Rachycentron canadum

A 4‐week feeding trial was conducted using five groups of hybrid grouper (61.15 ± 0.15 g) to explore the potential effects of three methionine (Met) sources. Five isonitrogenous and isolipidic diets were formulated that included a fishmeal (FM) diet; nonsupplemented (NS) diet; or NS diets with the addition of the L‐methionine (L‐Met), DL‐methionine (DL‐Met), or coated Met (Co‐Met) to obtain the same Met level as the FM diet. Fish were randomly distributed into sea cages (30 fish per cage). Weight gain and specific growth rate in the DL‐Met group and FM group were significantly higher than those in all other groups (p < .05). In the proximal and distal intestines, of the 20 gene and time combinations (10 per gene), there were 14 combinations (70%) in which there were no significant differences in gene expression levels between the FM and DL‐Met groups (p > .05). The main reason for the same growth effect between the DL‐Met and FM groups may be attributed to the synchronized absorption at most time points after feeding, which had similar expression patterns of B⁰AT1 and ASCT2 of the proximal and distal intestines between two groups.     

Effects of salinity on growth characteristics and osmoregulation of juvenile cobia,Rachycentron canadum (Linnaeus 1766), reared in potassium‐amended inland saline groundwater

The suitability of inland saline groundwater as a medium to culture juvenile cobia, Rachycentron canadum, was assessed. In the first experiment, juvenile cobia stocked in raw (unamended) saline groundwater at salinities of 5, 10, and 15 g/L exhibited complete mortality after 108, 176, and 195 hr, respectively. The second experiment evaluated the rearing of juvenile cobia (mean weight ~9.23 ± 0.12 g) in potassium (K⁺)‐amended saline groundwater (100% K⁺ fortified) and reconstituted seawater at salinities of 5, 10, and 15 g/L to assess growth and osmoregulation in distinct culture media. Following 60 days of culture, all fish survived the experimental period. Final mean bodyweight of cobia reared in K⁺‐amended saline groundwater (103.2–115.8 g) and seawater (111.2–113.8 g) of different salinities did not vary significantly (p > .05). No differences (p > .05) were observed in specific growth rate, weight gain (%), and feed conversion ratio between treatment groups. Serum osmolality increased with salinity and was significantly higher (p < .05) for fish in K⁺‐amended saline groundwater (353–361 mOsmol/Kg) than in reconstituted seawater (319–332 mOsmol/Kg), although differences were not observed between salinities by water type. Cobia stocked in saline groundwater of different salinities were osmoregulating normally, and the higher values observed may be because of variations in ionic composition and other interfering ions in saline groundwater. Trial results suggest that juvenile cobia can achieve optimal growth in K⁺‐amended saline groundwater of low and intermediate salinities.     

The role of dietary methionine concentrations on growth,metabolism and N‐retention in cobia (Rachycentron canadum) at elevated water temperatures

This study determined impacts of dietary methionine concentrations at two temperatures on growth, feeding efficiency and N‐metabolites in juvenile cobia. Methionine concentrations of the experimental diets were deficient (M9; 9 g/kg), sufficient (M12; 12 g/kg) and surplus (M16, 16 g/kg). Water temperature was normal (30°C) or elevated (34°C). Twenty cobia in triplicate tanks were fed the experimental diets for 6 weeks. Both methionine and temperature affected cobia's growth and feeding efficiency. Cobia fed M9 performed lower than the fish fed M12 and M16 diets. Additionally, cobia reared at 34°C performed poorer than at 30°C, probably due to lower voluntary feed intake in the fish reared at 34°C. Protein efficiency ratio and protein productive value in cobia fed M9 diet were less than M12 or M16 diets. This was confirmed with the improved retentions of indispensable amino acids (AAs). No interactions between methionine and temperature were observed in growth and protein accretion. At 30°C, CF improved, while HSI and VSI declined upon methionine supplementation levels. Of which an interaction between temperature and methionine was present. Plasma, muscle and liver free AA and N‐metabolites were affected by methionine and temperature. Furthermore, temperature affected cobia's lipid class composition, resulting in increased phospholipids and cholesterol at 34°C.     

Establishment,characterization, virus susceptibility and transfection of cell lines from cobia,Rachycentron canadum (L.), brain and fin

Establishment and characterization of two cobia, Rachycentron canadum, cell lines derived from cobia brain (CB) and cobia fin (CF) are described. Caudal fin and brain from juvenile cobia were dissociated for 30 and 10 min, respectively, in phosphate‐buffered saline containing 0.25% trypsin at 25 °C. The optimal culture condition for both dissociated cells (primary cell culture) was at 28 °C in Leibovitz‐15 medium containing 10% foetal bovine serum. The cells have been sub‐cultured at a ratio of 1:2 for more than 160 passages over a period of 3 years. Origin of the cultured cells was verified by comparison of their sequences of mitochondrial cytochrome oxidase subunit I genes (cox I) with the cox 1 sequence from cobia muscle tissue. The cell lines showed polyploidy. No mycoplasma contamination was detected. Susceptibility to grouper iridovirus was observed for the CB cell line but not the CF cell line. Both cell lines expressed green fluorescent protein after being transfected with green fluorescent reporter gene driven by the cytomegalovirus promoter.     

Effects of dietary carbohydrate sources on the growth performance and hepatic carbohydrate metabolic enzyme activities of juvenile cobia (Rachycentron canadum Linnaeus.)

An 8-week feeding trial was conducted to evaluate the effect of dietary carbohydrate sources on the growth performance and hepatic carbohydrate metabolic enzyme activities of juvenile cobia. Six experimental diets were formulated to contain 20% glucose, sucrose, maltose, dextrin, corn starch and wheat starch respectively. The results indicated that fish fed the wheat starch and dextrin diets showed significantly better weight gain, specific growth rate and protein efficiency ratio compared with those fed the other diets. However, fish fed the glucose diet had a significantly lower survival and condition factor than those fed the other diets. There were significant differences in the total plasma glucose and triglyceride concentration in fish fed diets with different dietary carbohydrate sources. Haematocrit, haemoglobin, red blood cell and leucocytes were significantly affected by the dietary carbohydrate sources. The activities of glucose-6-phosphate dehydrogenase (G6PD), 6-phosphofructokinase (PFK) and fructose-1,6-bisphosphatase (FBPase) were significantly affected by the dietary carbohydrate sources, while fish fed the glucose diet showed higher G6PD, PFK and FBPase activities than those fed the other diets. These data indicated that dextrin and wheat starch were the most optimal carbohydrate sources for juvenile cobia.     

军曹鱼幼鱼商品饲料中铁、锌适宜添加量的研究

在军曹鱼幼鱼商品饲料中添加铁100、200、300 mg/kg,添加锌30、110、190 mg/kg,共投喂56 d.试验结束时测定,军曹鱼成活率、质量增加率、红细胞含量、肌肉铁含量、骨骼铁、锌含量和肝脏铁、锌含量明显受饲料中铁、锌添加量的影响;血清碱性磷酸酶活性受铁、锌交互作用影响显著,饲料系数主要受铁锌交互作用的影响.以成活率和质量增加率为评价指标,军曹鱼幼鱼商品饲料中铁和锌适宜添加量分别为200 mg/kg和110 mg/kg.     

Genetic characteristics of Streptococcus dysgalactiae isolated from cage cultured cobia,Rachycentron canadum (L.)

Disease outbreaks occurred during 2007–2013 in Taiwan with 2.5–10% mortality among the cage cultured cobia, Rachycentron canadum (L.), characterized by the presence of polyserositis, pericarditis and peritonitis. The micro‐organisms isolated from internal organs were Gram‐positive cocci. The isolates were confirmed as Streptococcus dysgalactiae by a polymerase chain reaction assay that yielded the expected specific 259 bp amplicon. Additionally, partial sequence of the 16S–23S rDNA intergenic spacer region of the GCS strain isolates from fish was also compared and produced 100% sequence identity with S. dysgalactiae (GenBank accession number AB252398 ). The genetic characterization was then determined by pulsed‐field gel electrophoresis (PFGE) analysis. Based on PFGE, the Apa I or Sma I digestion patterns of chromosomal DNA of these isolates were grouped into three main clusters. Taiwanese strains were divided into two clusters, and the tet(M) gene was detected in cluster 1 (pulsotypes: A1–A2 and S1–S3), but not in cluster 2 strains (pulsotypes: A3–A4 and S4–S5). Three Japanese strains from amberjack, Seriola dumerili (Risso), were grouped into cluster 3 (pulsotypes: A5–A7 and S6–S8) and displayed no mortality to cobia in the challenge experiment. Conversely, Taiwanese strains from cobia and snubnose pompano, Trachinotus blochii (L.), displayed a mortality rate of 50–87.5% in cobia.     

军曹鱼MHC-Ⅱα基因全长cDNA的克隆及其组织表达分析

本研究根据其他鱼类MHC-Ⅱα基因的保守序列设计兼并引物,运用同源克隆和末端快速扩增方法扩增了军曹鱼MHC-Ⅱα基因的全长cDNA序列,并对cDNA及其氨基酸序列进行了分析,比较了军曹鱼和其他物种的MHC-Ⅱα氨基酸序列的差异,分析了军曹鱼MHC-Ⅱα基因的组织分布及经LPS刺激后头肾组织中MHC-Ⅱα基因的表达变化。结果表明,军曹鱼MHC-Ⅱα cDNA全长998 bp,包括53 bp的5′末端非编码区（5′UTR）、234 bp的3′末端非编码区（3′UTR）及711 bp的开放阅读框（ORF）,编码236个氮基酸,其蛋白质分子质量约为25.94 ku,等电点为4.39;军曹鱼MHC-Ⅱα蛋白质序列具有一些重要的特征,包括前导肽、α1、α2、CP/TM/CYT区和保守的半胱氨酸等;军曹鱼MHC-Ⅱα与鼠、人及其它鱼类的氨基酸同源性在25.0%～69.5%之间。Real-time PCR检测结果显示,MHC-Ⅱα基因在正常军曹鱼组织中均有表达,但其表达量在各种组织中存在差异,其中较强表达于头肾、鳃,中等程度表达于脾脏、肠,在心脏、脑、肌肉中表达较弱;经LPS刺激后,头肾中MHC-Ⅱα基因表达下调。     

Effect of Zataria multiflora Boiss (Avishan Shirazi) Essential Oil on Oxidative Progress in Frozen Cobia Fish Fillets During Storage

Antioxidants have been widely used as additives to provide protection against oxidative degradation of foods by free radicals. The effect of thyme essence (Zataria multiflora Boiss) on the rancidity development in cobia (Rachycentron canadum) fillets during frozen storage was studied. Cobia fillets were treated with thyme essence (250 and 500 ppm) and then stored at??18°C for up to 6 months. Rancidity development was measured by several biochemical indices including free fatty acids (FFA), peroxide value (PV), thiobarbituric acid (TBA), and complemented by sensory analysis (flesh odor, consistency, and appearance). Also, pH and expressible moisture were measured during 6-month storage. Proximate composition was also determined in the first day. TBA, PV, and FFA levels increased in all treatments due to lipid oxidation. Thyme essence showed an antioxidative effect in cobia fillets during frozen storage as indicated by TBA, PV, and FFA levels. Results showed that FFA, primary and secondary oxidation products, expressible moisture (EM), and pH of thyme essence treated samples were significantly lower than those of the control samples (p < 0.05). Thyme essence retarded oxidative changes in frozen cobia fillets, and the best oxidation inhibition was obtained using thyme essence at 500 ppm.