不同蛋白水平下添加抗菌肽对赤点石斑鱼影响的研究

评估了不同饲料蛋白水平(44%,48%,52%)和抗菌肽浓度(0.01%,0.02%)对赤点石斑鱼生长性能、血清生化、消化酶和抗氧化能力的影响。实验组分别为:D1(44%,0.01%),D2(48%,0.01%),D3(52%,0.01%),D4(44%,0.02%),D5(48%,0.02%),D6(52%,0.02%)。每组3个平行,养殖8周结果如下:一定范围内,增重率、特定生长率和饲料转化率随蛋白水平和抗菌肽浓度增加,D5组显著高于D1组(P0.05)。形态学指标,D6组肝体比(HSI)、脏体比(VSI)和肠系膜脂肪(IPF)显著低于D5组(P0.05)。同一蛋白水平下增加抗菌肽浓度可降低肌肉粗脂肪含量,D4组显著低于D1组(P0.05)。血清总蛋白、白蛋白含量随蛋白水平和抗菌肽浓度的增加而升高(P0.05),D3和D5组谷丙转氨酶、谷草转氨酶和血糖显著低于其他组(P0.05)。D5组胰蛋白酶高于其他组(P0.05),D1组过氧化物酶显著低于其他组(P0.05)。随着抗菌肽浓度增加,胃蛋白酶、过氧化物酶、过氧化氢酶活性增加,且丙二醛含量降低(P0.05)。实验结果表明,48%蛋白水平下添加0.02%抗菌肽可以提高赤点石斑鱼生长性能,增强消化酶活性和抗氧化能力,降低饲料系数。     

蛋氨酸钴对珍珠龙胆石斑鱼幼鱼生长、矿物元素沉积及肝脏酶活力的影响

在基础饲料中添加不同梯度的蛋氨酸钴(Co Met),制成钴含量分别为0.30 mg/kg、1.75 mg/kg、3.42 mg/kg、6.73 mg/kg、12.56 mg/kg、25.50 mg/kg的6组实验饲料(记作D1、D2、D3、D4、D5、D6组),饲喂初始体重(60.02±0.42)g的珍珠龙胆石斑鱼(Epinephelus lanceolatu♂×E.fuscoguttatus♀)幼鱼8周,研究蛋氨酸钴对其生长、矿物元素沉积和肝脏酶活力的影响。结果表明,随饲料中蛋氨酸钴含量的增加,实验鱼增重率(WGR)和特定生长率(SGR)均先升后降,D3组显著高于其他组(P0.05);饲料系数(FCR)先降后升,D3、D4组显著低于其他组(P0.05)。蛋氨酸钴对肌肉水分和灰分含量均无显著影响(P0.05);粗蛋白含量呈降低的趋势,D5、D6组显著低于D1~D4组(P0.05),但二者之间无显著差异(P0.05);粗脂肪含量先降后升,D4组显著低于其他组(P0.05),D6组显著高于其他组(P0.05)。肝脏、肠道、脊椎骨及肌肉钴含量均随饲料中蛋氨酸钴的增加而显著增加(P0.05);全鱼钴沉积率呈先升高后平稳的趋势,D1组显著低于其他组(P0.05)。肝脏铁、铜和锌含量随饲料中蛋氨酸钴的增加呈先升后降的趋势;肝脏锰含量显著降低(P0.05)。随饲料中蛋氨酸钴的增加,肝脏谷胱甘肽过氧化物酶(GSH-PX)活力先升后降,D5组显著高于其他组(P0.05);D5、D6组肝脏丙二醛(MDA)含量显著低于其他组(P0.05);肝脏精氨酸酶(DArg)活力随饲料中蛋氨酸钴的增加显著升高(P0.05);D4~D6组肝脏羧肽酶A(CPA)活力显著高于D1~D3组(P0.05),但三组之间无显著差异(P0.05)。本实验条件下,以SGR为评价指标,经折线回归分析,珍珠龙胆石斑鱼幼鱼对钴的最适需求量为3.25 mg/kg,即53.28 mg(Co Met)/kg(饲料)。     

不同蛋白水平和不同能量蛋白比饲料对点带石斑鱼生长的影响

以鱼粉和酪蛋白为蛋白源,添加鱼油、豆油、α-淀粉、纤维素以及适量的维生素和矿物质配制成试验饲料,蛋白质设置3个水平:41%、47%、53%,能量设置3个水平:1 256.1、1 465.5、1 674.8 kJ/100 g.经过59 d饲养点带石斑鱼幼鱼,以生长速度、净增重、增重率、蛋白质效率、饲料系数为评价指标,结果表明:点带石斑鱼幼鱼配合饲料中适宜的蛋白水平为47%,适宜的能量蛋白比为31.30～33.50 kJ/g蛋白.     

Genomic and phylogenetic characterization of Shewanella xiamenensis isolated from giant grouper (Epinephelus lanceolatus) in Taiwan

Shewanella xiamenensis is an emerging pathogen causing intra‐abdominal infection and intestinal colonization. Epidemiologic clues suggest its role as a potential food‐borne zoonotic agent. To date, four genome sequences of S. xiamenensis have been made publicly available. All of them were isolated from water samples. In this study, we characterized the genome of a S. xiamenensis strain isolated from a giant grouper in Taiwan. The genome of S. xiamenensis ZYW1 is 4,827,717 bp in length and encodes 4,239 open reading frames. Its genomic sequence shares high homology with other S. xiamenensis strains. bla_OXA‐416 was identified. This is the first detection of S. xiamenensis in Taiwan. These genomic data and analyses contribute to our understanding of S. xiamenensis and may help to elucidate disease‐causing mechanisms in future studies.     

斜带石斑鱼甘露糖受体基因的克隆表达及其功能

为了研究斜带石斑鱼甘露糖受体(Epinephelus coioides mannose receptor, Ec MR)在抗赤点石斑鱼神经坏死病毒(red-spotted grouper nervous necrosis virus,RGNNV)感染中的免疫功能,实验成功克隆与表达了EcMR。结果显示,EcMR cDNA全长4 793 bp,共编码1 446个氨基酸。Ec MR的蛋白结构域包括1个信号肽(signal peptide)、1个富含蓖麻类β型三叶草结构域(RICIN)、1个Ⅱ型纤维连接蛋白结构域(FNⅡ)、8个串联的C-型凝集素样结构域(CLECTs)以及1个跨膜结构域(transmembrane region)。实时定量PCR(qRTPCR)和细胞免疫荧光(IF)分析结果显示,Ec MR在斜带石斑鱼的8个组织中均有表达,其mRNA的相对表达量顺序为鳃头肾脑脾脏肝脏外周血心脏肌肉。在研究Ec MR是否参与RGNNV入侵过程中时发现,RGNNV可以在GF-1细胞系中快速增殖,同时显著激活Ec MR的表达。为进一步探究RGNNV对GF-1细胞系的影响,本实验通过双染色法检测了RGNNV感染GF-1细胞系后的细胞凋亡情况,研究表明,RGNNV感染可以促进GF-1细胞系的凋亡,并且细胞凋亡率随着RGNNV感染时间延长而增加。同时,qRT-PCR和酶活性检测结果显示,RGNNV的感染可以显著促进凋亡相关基因的转录水平以及Caspase-3、Caspase-8、Caspase-9的酶活性水平。综上所述,本研究成功克隆了EcMR,并揭示了其在RGNNV入侵过程中一定的相关性。本研究结果将为石斑鱼病毒性疾病的防控提供参考。     

驼背鲈线粒体细胞色素b基因的序列分析

采用PCR—DNA测序技术，对驼背鲈线粒体DNA细胞色素b（Cytb）基因全序列进行分析。获得长度为1141bp的Cytb基因片段，序列碱基组成平均是T为29．1％，C为32．3％，A为23．2％，G为14．0％，A＋T的碱基百分比总体上略大于C＋G，平均值分别是52．3％和46．3％。与其它鱼类相同基因片段碱基序列含量相似。在由核苷酸推导的氨基酸中，亮氨酸（Leu）所占的比例最高，为16．03％。     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

Effects of dietary live yeast Debaryomyces hansenii on the immune and antioxidant system in juvenile leopard grouper Mycteroperca rosacea exposed to stress

This study investigated the effects of the dietary intake of live yeast Debaryomyces hansenii (CBS 8339) on the immune response of juvenile leopard groupers Mycteroperca rosacea and on their resistance to infection by dinoflagellates Amyloodinium ocellatum.During 4 weeks, juvenile groupers were fed either a diet supplemented with D. hansenii at 10⁶ cfu g^− 1 or a control diet. Both groups were then transferred to a system infected with A. ocellatum and maintained in this stressful environment, for one week. Rearing conditions were favorable to the proliferation of the dinoflagellate and, at 7 days post-infection, the cumulative mortality rate of the control group was 89.5%, suggesting that the yeast-enriched diet conferred protection against the parasite. Only fish with yeast-supplemented diet recovered from the A. ocellatum challenge.The immunological response of both groups of fish was evaluated. Samples were collected at weeks 4 (stressed fish) and 7 (recovering fish) to determine the levels of hemoglobin, plasmatic protein, and immunoglobulin M (IgM) as well as the activity of antioxidant enzymes (SOD and CAT). IgM increased significantly (P ≤ 0.05) in the recovering fish, which had all been fed the yeast diet. A significantly increase was also observed in superoxide dismutase activity in juvenile fish fed the yeast diet; however catalase activity was not affected by the dietary yeast supplement. Overall, fish that received the yeast supplement in their diet have an enhanced immune response as compared with their control counterparts. The present results show that a diet supplemented with D. hansenii stimulates the immune system of juvenile M. rosacea thus enhancing their resistance against A. ocellatum.     

短期饥饿对杂交石斑鱼 (Epinephelus fuscogutatus♀ ×E.polyphekadion♂)幼鱼消化酶活性与血清生化指标的影响

为了解短期饥饿对杂交石斑鱼(Epinephelus fuscogutatus♀×E.polyphhekadion♂)幼鱼消化酶活性和血清生理生化指标的影响,对杂交石斑鱼幼鱼进行短期饥饿处理(0 d、5 d、10 d、15 d),分别为S0组、S1组、S2组和S3组,并以S0组为对照组。结果表明,随着饥饿时间的延长,各种消化酶活性呈下降趋势,淀粉酶活性呈先上升后下降趋势。血清碳水化合物、胆固醇、甘油三酯、总蛋白、白蛋白等生化指标在饥饿前期显著降低(P<0.05),随后维持在相对稳定的水平。AST、ALT、ALP活性呈“下降-上升-下降”趋势。分析表明,在短期饥饿过程中,杂交石斑鱼幼鱼早期主要消耗碳水化合物和脂肪物质,后期主要以蛋白质代谢能量来维持生命活动,短期饥饿并没有对杂交石斑鱼造成不可逆转的肝脏损伤。