Molecular divergence and application of the ITS-5.8S rDNA and RUBISCO spacer in <Emphasis Type="Italic">Porphyra haitanensis</Emphasis> Chang et Zheng (Bangiales,Rhodophyta)

To select a reliable and sensitive method for discriminating strains of Porphyra haitanensis, the nucleotide sequence of the internal transcribed spacer 1 to internal transcribed spacer 2 regions (ITS-5.8S) of nuclear ribosomal DNA and the intergenic spacer region of RUBISCO were compared in five wild and five cultivated Porphyra haitanensis strains. Based on molecular analyses, sequences of ITS-5.8S (about 1,210 bp) could be divided into three regions: ITS1, 5.8S, and ITS2. The ITS1 and ITS2 sequences of each strain differed, even between individuals collected from the same site. In contrast, 5.8S rDNA and RUBISCO spacer sequences were identical among the ten P. haitanensis strains, although differences were found among different Porphyra species. Phylogenetic analysis also supported these conclusions. These sequence features of highly conserved regions and diversified regions that occurred repeatedly in ITS-5.8S could be useful in discriminating germplasm of P. haitanensis strains or Porphyra species. In contrast, the RUBISCO spacer is only suitable for identifying Porphyra species. New coupled primers were designed to amplify only the 5.8S rDNA and ITS2 region of Porphyra. The sequences of these amplified fragments can be readily used to identify germplasm or to perform phylogenetic analysis of Porphyra spp.     

坛紫菜有丝分裂阻滞缺陷蛋白2基因(Ph-mad2)的克隆及其在单性生殖过程中的表达

为探究坛紫菜雌配子体单性生殖过程的细胞二倍化机制,实验基于坛紫菜雌性配子体的转录组信息,克隆和分析了坛紫菜的有丝分裂阻滞缺陷蛋白2(PhMAD2)基因(Phmad2),同时对其在坛紫菜单性生殖发生过程中处于不同发育时期的细胞中的表达特征进行了初步研究。结果显示,坛紫菜Ph-mad2基因的cDNA的完整开放阅读框为672 bp,编码223个氨基酸,分子量为23.8 ku。从氨基酸序列比对结果可知,坛紫菜PhMAD2蛋白中所具有的典型的HORMA结构域和保守的丝氨酸和苏氨酸磷酸化位点,均与团藻和莱茵衣藻MAD2蛋白相同。系统发育分析表明,坛紫菜与藻状菌纲的异丝水霉和寄生水霉的亲缘关系最近。实时荧光定量PCR检测显示,与可进行正常有丝分裂的营养细胞相比,在坛紫菜单性生殖过程中发生细胞二倍化的生殖细胞和单性孢子时期,Ph-mad2基因的表达量显著下降;但是,在随后可进行正常有丝分裂的单性孢子萌发体细胞和处于营养生长期的单性孢子体细胞中则表达上调。这一结果暗示Ph-mad2基因的表达下调可能阻碍了坛紫菜单性生殖初期单倍体细胞进行有丝分裂时纺锤体组装检验点的形成,在一定程度上致使它们发生了染色体加倍。     

Selection and characterization of an improved strain (ST‐2) of Pyropia haitanensis by 60Co‐γ ray irradiation

Although the improved strain HR‐5 of Pyropia haitanensis has improvement in yield and quality, it shows earlier maturation and shortened growth period. In this study, blades of HR‐5 strain were treated with ⁶⁰Co‐γ ray irradiation, and a mutant strain ST‐2 was selected and characterized. The blade length of ST‐2 strain was 401.4 cm on day 70 post‐culture, which was 1.9‐fold longer than that of the parental strain HR‐5. The blades of HR‐5 strain matured earlier and most were matured on day 55, while the ST‐2 strain was not matured until day 90. When blades of HR‐5 and ST‐2 strains were firstly cultured at 23°C for 40 days and their blade discs cultured at 29°C for another 30 days, the mean length of ST‐2 strain was 2.0‐fold longer than that of HR‐5 strain. Besides, the contents of Chl. a and phycobiliprotein and the conchospore numbers in ST‐2 were comparable to that in HR‐5 strain. In summary, compared with HR‐5, we observed delayed maturation, increased length, weight and high‐temperature resistance of the blades, high content of photosynthetic pigments, and high production of conchospores in ST‐2, suggesting it might be a better strain for large‐scale production.     

Influence of dietary zinc on lipid peroxidation,protein oxidation and antioxidant defence of juvenile Jian carp (Cyprinus carpio var. Jian)

This study was conducted to study the effects of dietary zinc on lipid peroxidation, protein oxidation and antioxidant defence of juvenile Jian carp (Cyprinus carpio var. Jian) by feeding fish with increasing levels of zinc (15.3, 26.9, 40.8, 58.2, 68.9 and 92.5 mg Zn kg^?1) for 6 weeks. Results indicated that malondialdehyde (MDA) content and protein carbonyls (PC) in serum were the highest in fish fed diet containing 15.3 mg zinc kg^?1 diet (P < 0.05). Serum antisuperoxide anion (ASA), superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px), glutathione reductase (GR) activities and glutathione (GSH) content were improved with increasing dietary zinc levels up to 40.8 mg zinc kg^?1 diet (P < 0.05) and levelled off (P > 0.05). Serum antihydroxy radical (AHR), catalase (CAT) and glutathione‐S‐transferase (GST) activities followed the similar pattern to that observed in ASA. The MDA and PC levels, ASA, AHR, SOD, CAT, GPx, GR, GST activities and GSH content in intestine, hepatosomatic and muscle tissue followed the similar pattern to that observed in serum. The present results indicated that zinc decreased lipid peroxidation and protein oxidation and improved antioxidant defence in fish.     

坛紫菜叶状体的抗生素联合除菌效果

为研究坛紫菜及其外生菌群对氯霉素、竹桃霉素和头孢噻肟3种抗生素的敏感性,通过将其与氨苄青霉素、卡那霉素和庆大霉素的组合,优化坛紫菜叶状体的除菌方法,并利用涂布平板、qRT-PCR及16S rRNA测序分析其除菌效果。结果显示,当坛紫菜先以氨苄青霉素(300 mg/L)、卡那霉素(100 mg/L)和庆大霉素(100 mg/L)混合处理18 h,再以氯霉素(50 mg/L)、头孢噻肟(200 mg/L)和竹桃霉素(50 mg/L)混合处理4 d,叶状体的健康率保持在96.3%以上,而对可培养细菌的抑制率达到99.9%。根据qRT-PCR和16S rRNA的结果可知,该种组合方法较前3种抗生素的组合,其菌总数下降41.5%,微生物的丰度和多样性指数明显下降。其中,假单胞菌、交替赤杆菌、交替单胞菌以及海杆菌等得到针对性的抑制。研究表明,多种抗生素的优化组合能够对坛紫菜叶片的菌群结构产生显著影响,使其受到抑制,但仍无法达到绝对除菌的效果。     

几种经济红藻中茉莉酸合成途径的关键物质

为研究几种经济红藻中茉莉酸合成途径的关键物质,实验采用液相色谱—质谱联用技术(LC-MS)对茉莉酸生物合成途径相关物质建立检测方法,并对龙须菜、坛紫菜受到机械损伤时各物质的变化情况进行检测。以100%甲醇提取茉莉酸(JA)、茉莉酸甲酯(MeJA)、12-氧-植物二烯酸(12-OPDA)和13-氢过氧化亚麻酸(13-HpOTE),利用XBridge TM C18色谱柱,以甲醇/水为流动相分离4种物质。优化条件下4种物质得到良好分离,加标回收率为81.23%～90.25%,检测限为0.04～0.56 ng/mL,灵敏度高。坛紫菜、龙须菜和真江蓠3种红藻中,坛紫菜中未检测到JA和13-HpOTE,4种物质均在另外2种藻中检测到。龙须菜受机械损伤胁迫后,4种物质在短时间内得到积累,其中13-HpOTE响应迅速。研究表明,红藻中可能存在类似于植物的茉莉酸合成途径,并参与对损伤的胁迫响应。     

Cytogenetic studies of Atlantic mojarras (Perciformes – Gerreidae): chromosomal mapping of 5S and 18S ribosomal genes using double FISH

Fish of the family Gerreidae, mainly species of the genera Diapterus and Eugerres, have high potential for cultivation, because of their saline tolerance. A detailed cytogenetic analysis of Diapterus auratus, Diapterus rhombeus and Eugerres brasilianus was conducted using conventional staining, C‐banding, Ag‐NOR, AT/GC‐specific fluorochrome staining and mapping of ribosomal sequences with 5S and 18S rDNA probes. All the species exhibited symmetrical karyotype, 2n = 48 acrocentric chromosomes. Ag‐NORs and 18S rDNA are present in the interstitial position on pair 1 (genus Diapterus) and pair 6 in Eugerres brasilianus. The 5S rDNA sites, located in the interstitial position (pair 11), are conserved in the three species. Heterochromatic regions are similar in the Diapterus species, showing a pattern of reduced and centromeric bands, differing from E. brasilianus, where, in addition to these, more prominent interstitial bands were observed. GC‐rich regions are located at ribosomal sites. Karyotypic comparison between Diapterus and Eugerres reveals similarity in chromosomal macrostructure, differing in C‐positive heterochromatin distribution and position of 18S sites, indicating the occurrence of structural microrearrangements. Although complementary analyses are needed, the similarities observed for these and other species suggest the possibility of breaking postzygotic barriers and their potential use, through induced interspecific or intergeneric hybridizations.     

Algal density alleviates the elevated CO2‐caused reduction on growth of Porphyra haitanensis (Bangiales,Rhodophyta), a species farmed in China

Growing of Pyropia haitanensis, a commercially farmed macroalga, usually increases their densities greatly during cultivation in natural habitats. To explore how the increased algal densities affect their photosynthetic responses to rising CO₂, we compared the growth, cell components and photosynthesis of the thalli of P. haitanensis under a matrix of pCO₂ levels (ambient CO₂, 400 ppm; elevated CO₂, 1,000 ppm) and biomass densities [low, 1.0 g fresh weight (FW) L^?1; medium, 2.0 g FW L^?1; high, 4.0 g FW L^?1]. Under ambient CO₂, the relative growth rate (RGR) was 5.87% d^?1, 2.32% d^?1 and 1.51% d^?1 in low, medium and high densities, and elevated CO₂ reduced the RGR by 27%, 25% and 12% respectively. Maximal photochemical quantum yield of photosystem II (F_V/F_M) was higher in low than in high densities, so were the light‐utilized efficiency (α), saturation irradiance (E_K) and maximum relative electron transfer rate (rETR_max). Elevated CO₂ enhanced the F_V/F_M in low density but not in higher densities, as well as the α, E_K and rETR_max. In addition, elevated CO₂ reduced the content of chlorophyll a and enhanced that of carotenoids, but unaffected phycoerythrin, phycocyanin and soluble proteins. Our results indicate that the increased algal densities reduced both the growth and the photosynthesis of P. haitanensis and alleviated the elevated CO₂‐induced negative impact on growth and positive impact on photosynthesis. Moreover, the elevated CO₂‐induced reduction on growth and promotion on photosynthesis indicates that rising CO₂ may enhance the loss of photosynthetic products of P. haitanensis through releasing organic matters.     

细胞骨架和粘性多糖对坛紫菜壳孢子极性形成的影响

应用解聚微丝和微管细胞骨架的特异性药物,结合组织化学染色方法观察了坛紫菜壳孢子早期发育阶段中的细胞极性形成过程。观察结果表明,坛紫菜壳孢子从放散入水作变形运动到萌发成二细胞的孢子苗共有两次细胞极性形成过程:第一次发生在坛紫菜壳孢子作变形运动期间(形成前后轴);第二次发生在壳孢子拉长呈椭圆形的一细胞萌发体期间(形成细胞的上下轴)。在这两次细胞极性形成过程中,壳孢子外的粘性多糖,尤其是硫酸化的酸性粘多糖主要聚集在运动壳孢子的后缘和壳孢子萌发体的下方,说明粘性多糖有协同形成、稳定壳孢子细胞极性的作用。刚放散的壳孢子经高浓度的细胞骨架抑制剂处理后,壳孢子的正常运动被阻止,从而影响壳孢子的第一次细胞极性形成,导致壳孢子不能萌发。运动后的壳孢子经高浓度的抑制剂处理后,出现了假根形成和细胞分裂面不正常的畸形萌发体,它们的基部均没有粘性多糖的显色。     

Molecular diagnosis of Myxobolus spirosulcatus associated with encephalomyelitis of cultured yellowtail, Seriola quinqueradiata Temminck & Schlegel

Mass mortality of cultured yellowtail, Seriola quinqueradiata, has recently been reported from fish farms in western Japan. Previous studies revealed that diseased fish were characterized by encephalomyelitis and presporogonic stages of a myxosporean‐like parasite in the spinal cord. However, the parasite has remained unidentified because of the lack of mature stages being present. Thus, in the present study, analysis of the small subunit ribosomal DNA (18S rDNA) of the parasite as well as in situ hybridization (ISH) studies using histological sections of the infected tissue was conducted. The 18S rDNA of the myxosporean had higher sequence similarities with those of bile‐duct‐infecting myxosporeans rather than those infecting nervous tissues and was identified as Myxobolus spirosulcatus. The ISH using specific probes demonstrated that the DNA amplified was derived from the multinuclear organisms found in histological sections. A highly sensitive and specific PCR‐based assay for M. spirosulcatus was developed, which revealed a high prevalence of infection in cultured yellowtail that exhibited the clinical signs of encephalomyelitis.     

Identification and real‐time qPCR quantification of a nitrite‐N degrading bacterial strain in aquatic water

Biological nitrogen removal technology using microbe is an efficient process for nitrogen removal from aquatic water. To determine the key of efficient application on how the effect of probiotics lasting, a nitrite nitrogen (nitrite‐N) degrading 8DO17 strain Pseudomonas was screened and a method for quantification was explored. Real‐time qPCR assays based on the 16S rRNA genes (16S rDNA) were used to quantify the probiotics. The results showed that (i) the nitrite‐N degradation rate of the 8DO17 strain was 99%; (ii) a wide spectrum of pH (7.0–9.0) and concentration of nitrite‐N (0.5–10 mg L^?1) and the highest rate of nitrite‐N degradation near to 100% under optimum conditions (35°C, salinity 30, pH 7.5) was explored; (iii) no significant differences were found in the survival, total haemocyte count, antibacterial activities and bacteriolytic activity of the shrimp between the treatments and the control (P > 0.05); (iv) for the use of real‐time PCR based on 16S rDNA test, detection limit is 10³ DNA copies and an excellent correlation coefficients (R² = 0.999) was obtained; and (v) in the application of real‐time PCR assay in four nitrite‐N degrading samples, highly significant positive correlation relation (P < 0.01) was found between log copy number of 16S rDNA and the rate of nitrite‐N degradation. The results suggest the real‐time PCR is a very rapid and sensitive technique for monitoring the dynamic changes and assessing the effect in practical application of the 8DO17 strain in aquatic water.     

Dietary phosphorus prevents oxidative damage and increases antioxidant enzyme activities in intestine and hepatopancreas of juvenile Jian carp

This study was conducted to investigate the effect of dietary phosphorus on the intestine and hepatopancreas antioxidant capacity of juvenile Jian carp (Cyprinus carpio var. Jian). Jian carp, with an average initial weight of 7.17 ± 0.01 g, were fed with diets containing graded concentrations of available phosphorus, namely 1.7 (control), 3.6, 5.5, 7.3, 9.2 and 11.0 g kg^?1 diet for 9 weeks. Results showed that, in intestine and hepatopancreas, content of malondialdehyde (MDA), protein carbonyl (PC) and glutathione (GSH), capacity of anti‐superoxide anion (ASA) and anti‐hydroxyl radical (AHR), and glutathione reductase (GR), catalase (CAT), glutathione S‐transferase (GST), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were significantly affected by dietary phosphorus levels (P < 0.05). Regression analysis showed that significant quadratic responses occurred in MDA content and ASA, GST, GPx and AHR activities in intestine, GSH content and CAT and SOD activities in hepatopancreas (P < 0.05). These results indicate that optimal level of dietary phosphorus prevented oxidative damage and increased antioxidant enzyme activities in the intestine and hepatopancreas of juvenile Jian carp. The phosphorus requirement estimated from MDA using quadratic regression analysis was 5.7 g kg^?1 diet.     

Effects of dietary phosphorus and starch levels on growth performance,body composition and nutrient utilization of grass carp (Ctenopharyngodon idella Val.)

Six iso‐nitrogenous (410 g kg^?1) diets with three levels of total phosphorus (P4, P10 and P18 g kg^?1) and two levels of starch (S200 and S350 g kg^?1) were fed to triplicate groups of 30 fish to evaluate whether the high level of dietary phosphorus could improve the utilization of starch. Over 8‐week‐growth trial, best weight gain (WG) and specific growth rate (SGR) (P < 0.05) were observed in fish fed the P10/S200 and P18/S200 diets. WG and SGR significantly decreased as starch levels increased whereas for P4, while lipid contents of liver and whole body, hepatosomatic index and intraperitoneal fat ratio (IPF) significantly increased. These results suggested that high dietary starch will depress the growth performance and cause lipid accumulation. Within both starch levels, fish fed diet with P4 tended to produce lower (P < 0.05) WG and SGR, and had higher (P < 0.05) values of IPF. The whole body lipid, ash, calcium, phosphorus and iron contents were significantly affected by dietary phosphorus levels. Supplied phosphorus could improve the growth and decrease the whole body lipid, but there is no more effect after the phosphorus requirement was met at 10 g kg^?1.     

Mycobacterium salmoniphilum infection in farmed Atlantic salmon, Salmo salar L

Multiple greyish‐white visceral nodules containing abundant rapidly growing and acid‐fast bacteria, subsequently identified as Mycobacterium salmoniphilum, were detected in moribund and newly dead market‐sized fish during a period of increased mortality in an Atlantic salmon, Salmo salar, farm in western Norway. Isolates cultured from diseased fish were phenotypically consistent with Mycobacterium sp. previously isolated from Atlantic salmon [MT 1890 (= NCIMB13533), MT1892, MT1900 and MT1901] in the Shetland Isles, Scotland. Partial sequences of 16S rDNA, ribosomal RNA internal transcribed spacer (ITS1), 65‐kDa heat‐shock protein (Hsp65) and β subunit of RNA polymerase (rpoB) revealed 97‐99% similarity with M. salmoniphilum type strain ATCC 13758^T. The source of infection was not confirmed. Koch’s postulates were fulfilled following experimental challenge of Atlantic salmon with field isolate NVI6598 ( FJ616988 ). Mortality was recorded in experimentally infected fish; however, the infection remained subclinical in the majority of affected fish over the 131‐day challenge period.     

A new primer for 16S rDNA analysis of microbial communities associated with <Emphasis Type="Italic">Porphyra yezoensis</Emphasis>

To construct high-quality 16S rDNA clone libraries for microbial communities associated with Porphyra yezoensis and to minimize the detection of rDNA from leafy gametophytes of P. yezoensis, we designed a new 16S rDNA universal primer (75F). Of the clones prepared using 75F, which was designed to distinguish between bacteria and P. yezoensis, 95% were classified into four groups, namely, β-proteobacteria, γ-proteobacteria, Lentisphaerae, and Flavobacteria. PCR-based analysis of the 16S rDNA primer constructed in this study can be used to implement 16S rDNA-based methodologies for the investigation of microbial community composition and diversity related to the Porphyra group.     

Evaluations of lactic acid bacteria as probiotics for juvenile seabass Lates calcarifer

Lactic acid bacteria (LAB) were isolated from adult, wild‐caught and farmed seabass (Lates calcarifer) intestines for evaluation as possible probiotics using the well agar diffusion method. Five LAB isolates (designated as LAB‐1–5) were found to inhibit Aeromonas hydrophila, a known seabass pathogen. Median lethal concentrations (LC₅₀) of A. hydrophila on juvenile seabass were measured in aquaria. Median lethal concentration values of 7.76, 7.47 and 7.26 log₁₀ CFU mL^?1 for 72, 96 and 120 h, respectively, were found. Juvenile seabass (0.6±0.2 g) were cultured in aquaria and fed individual LAB‐1–5 fortified feeds with 7 log₁₀ CFU g^?1 LAB. Seabass fed LAB‐4 fortified feed had significantly greater growth (P<0.05) than fish fed other feeds. Seabass fed LAB‐4 also had greater survival, but this was non‐significant (P<0.05). Challenge tests of LAB‐4 fed seabass with A. hydrophila at ～7 log₁₀CFU mL^?1 yielded significantly greater survival compared with control seabass (P<0.05). Aeromonas hydrophila infections in seabass were confirmed by observing disease manifestation and by immunohistochemistry techniques. LAB‐4 was preliminarily identified using lactic acid analysis, biochemical and physical characteristics. It was further identified using 16S rDNA sequencing. LAB‐4 was identified as Weissella confusa (identity of 99%). GenBank accession number for the 16S rDNA sequence for LAB‐4 was AB023241.     

Development of primers and a procedure for specific identification of the diatom <Emphasis Type="Italic">Thalassiosira weissflogii</Emphasis>

A recent study showed Thalassiosira weissflogii to be a diatom containing suitable nutrition for larviculture of the black tiger shrimp, Penaeus monodon. Accurate and practical identification of this diatom species is therefore important for commercial hatcheries. The purpose of this study was to establish a DNA-based method of identification to supplement morphological examination, avoiding confusion with other Thalassiosira sp. Primers, 18SF/28SR1, specific for ribosomal DNA genes (3′-end of 18S rDNA through 5′-end of 28S rDNA, covering two internal transcribed spacers), were employed as a first-step polymerase chain reaction, followed by a second nested amplification using specifically designed primers, ITS1-F-D/ITS1-R-D. The nested-PCR result revealed specificity in the detection, distinguishing T. weissflogii from T. pseudonana, Cyclotella meneghiniana, and Chaetoceros sp., and the PCR fragment of the amplified region had a sequence that was 99% identical to the T. weissflogii sequence held by GenBank.     

Methionine hydroxy analogue prevents oxidative damage and improves antioxidant status of intestine and hepatopancreas for juvenile Jian carp (Cyprinus carpio var. Jian)

Oxidative damage and antioxidant status of intestine and hepatopancreas for juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of methionine hydroxy analogue (MHA: 0, 5.1, 7.6, 10.2, 12.7, 15.3 g kg^?1 diet) for 60 days were studied. Radical scavenging ability, antioxidant enzymes activities such as superoxide dismutase (SOD), catalase (CAT), glutathione‐S‐transferase (GST), glutathione peroxidase (GPX) and glutathione reducase (GR), as well as glutathione (GSH), protein carbonyl (PC) and malondialdehyde (MDA) contents were assayed in these tissues. Results indicated that anti‐superoxide anion capacity in intestine and anti‐hydroxyl radical capacity in hepatopancreas significantly improved with dietary MHA levels up to 7.6 and 10.2 g kg^?1 diet respectively, whereupon they decreased (P < 0.05). SOD, CAT, GST, GPX, GR activities in intestine and hepatopancreas, as well as GSH content in hepatopancreas significantly increased with optimal MHA levels which were in the range of 5.1–10.2 g kg^?1 diet, and thereafter decreased (P < 0.05). Meanwhile, MDA and PC contents in these tissues together with GOT and GPT activities in plasma significantly decreased with optimal MHA levels which were in the range of 5.1–7.6 g kg^?1 diet, and thereafter increased (P < 0.05). These results suggested that MHA improved antioxidant status and depressed lipid and protein oxidation in intestine and hepatopancreas.     

Influence of dietary phosphorus levels on growth,body composition,metabolic response and antioxidant capacity of juvenile snakehead (Channa argus × Channa maculata)

A study was conducted to estimate the optimum requirement of dietary phosphorus (P) for Channa argus × Channa maculata. Effects of dietary P levels on the tissue composition, serum biochemical parameters and antioxidant status were also examined. Five practical diets were formulated to contain graded levels (4.8 g kg^?1, 6.4 g kg^?1, 7.9 g kg^?1, 9.4 g kg^?1 and 11.0 g kg^?1) of available P from dietary ingredients and monocalcium phosphate (MCP). Each diet was randomly assigned to triplicate groups of 30 juvenile fish (initial body weight, 20.50 ± 0.53 g) for 8 weeks. The results showed that the specific growth rate (SGR) and weight gain (WG) were all significantly improved by dietary P up to 9.4 g kg^?1 (P < 0.05) and then levelled off beyond this level. Broken‐line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 9.6 g kg^?1. With the increase in dietary P level, protein efficiency rate (PER) increased significantly and reached a plateau, while the feed conversion ratio (FCR), the mesenteric lipid somatic index (MSI) and the whole‐body lipid content significantly reduced (P < 0.05). Dietary P levels also affected the mineralization (ash and P) of whole body, vertebrae and scale (P < 0.05). Quadratic analysis based on P contents in whole body, vertebrae, scale and ash content in vertebra indicated that the available P requirements were 10.4, 9.8, 10.0 and 10.3 g kg^?1, respectively. However, no differences were found in the whole‐body moisture, crude protein, serum calcium (Ca) contents or Ca/P value, as well as the viscerosomatic index (VSI) and hepatosomatic index (HSI) among all the treatments (P > 0.05). Triglyceride (TG), total cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C) and low‐density lipoprotein cholesterol (LDL‐C) decreased significantly, while serum P content, HDL‐C/TC and HDL‐C/LDL‐C value increased significantly with dietary available P levels (P < 0.05). No significant changes in superoxide dismutase activity and malondialdehyde (MDA) content were observed (P > 0.05), but serum catalase (CAT) and glutathione peroxidase (GPx) activities and the ratio of CAT/SOD and GPx/SOD increased significantly with increasing dietary P levels (P < 0.05). In conclusion, the optimal P requirement of juvenile snakehead in practical feed was 9.6 g kg^?1. Signs of P deficiency were characterized by poor growth, slightly reduced mineralization and the antioxidant capacity and an increase in body lipid content.     

Effect of dietary supplemental l‐carnitine on growth performance,body composition and antioxidant status in juvenile black sea bream,Sparus macrocephalus

Effects of dietary l ‐carnitine were studied in juvenile black sea bream (Sparus macrocephalus). The semipurified basal diet [crude protein 450 g kg^?1 dry matter (DM); crude lipid 126 g kg^?1 DM] was formulated to choose white fishmeal as the protein source and fish oil plus corn oil (1 : 1) as the lipid source. Six diets (control + diets 1–5) containing 0.1, 0.12, 0.16, 0.24, 0.39 and 1.1 g of l ‐carnitine kg^?1 diet were fed to triplicate groups of black sea bream (initial weight 13.10 ± 0.05 g) for 8 weeks. At the end of the feeding trial, growth performance, body composition and antioxidant status were determined. The results showed that relative growth rate (RGR) was significantly improved by the elevation of dietary l ‐carnitine level from 0.1 to 0.24 g kg^?1, but decreased with further increment (P < 0.05). Lipid content decreased significantly (P < 0.05) in the dorsal muscle whereas increased (P < 0.05) in the liver with the addition of dietary l ‐carnitine. Dietary l ‐carnitine supplements elevated enzymatic antioxidants (superoxide dismutase, SOD; catalase, CAT; glutathione‐S‐transferase, GST) activities (P < 0.05) yet decreased the content of non‐enzymatic factor, total sulphydryl groups (TSH) (P < 0.05). In summary, the optimum dietary l ‐carnitine level was 0.284 g kg^?1 diet by second‐polynomial regression analysis based on RGR (y = ?647.4x² +367.97x + 234.55; R² = 0.977, x = dietary l ‐carnitine levels, y = RGR), and dietary l ‐carnitine addition within the levels adopted in our study could depress lipid peroxidation in tissues of juvenile black sea bream.