Dietary protein requirement of juvenile obscure puffer,Takifugu obscurus

A 10‐week feeding experiment was conducted to determine the optimum dietary protein requirement of juvenile obscure puffer (Takifugu obscurus). Six isoenergetic (20 MJ kg^?1 gross energy) diets were formulated to contain graded levels of 34%, 38%, 42%, 46%, 50% or 54% crude protein (as dry matter basis). The results showed final body weight, weight gain and specific growth rate (SGR) increased significantly with increasing protein levels up to 42% and then decreased thereafer. Second‐order polynomial regression analysis (y = ?0.0024x² + 0.1788x ? 1.3196, R² = 0.9032) indicated a maximum SGR at protein level of 37%. Feed conversion ratio (FCR) decreased with increasing levels of dietary protein up to 42% and increased thereafter. Second‐order polynomial regression analysis (y = 0.0054x² ? 0.4351x + 10.391, R² = 0.753) indicated a minimum FCR at protein level of 40%. Protein efficiency ratio (PER) of fish fed the 34%, 38% and 42% diets was significantly higher than that of fish fed the 46%, 50% and 54% diets, and broken‐line analysis indicated PER tended to decrease when dietary protein level was higher than 40%. Generally, whole body lipid content, total cholesterol, low‐density lipoprotein cholesterol and triacylglycerol decreased with increasing levels of dietary protein. Fish fed the 42% protein diet showed the highest essential amino acids (histidine, isoleucine, leucine, lysine and threonine) and non‐essential amino acids (aspartic acid and glutamic acid) in muscle. Based on the second‐degree polynomial regression analysis of SGR and FCR and broken‐line analysis of PER, the optimal dietary protein level of obscure puffer is estimated to be between 37% and 40% (% as dry matter basis).     

条石鲷(Oplegnathus fasciatus)源美人鱼发光杆菌杀鱼亚种(Photobacterium damselae subsp. piscicida)的分离鉴定

2013年浙江省舟山市某网箱养殖条石鲷(Oplegnathus fasciatus)暴发了一种严重的疾病,病鱼主要症状为脾、肾出现1-2 mm的白色类结节.从患病鱼内脏处分离得到1株优势菌OF-1,经人工感染实验证实为此次引起条石鲷死亡的致病菌,半数致死量为5.93×104 CFU/g.形态学观察结果显示,菌株OF-1为革兰氏阴性、短杆状,在TCBS培养基上不生长.API 20E细菌鉴定系统、16S rRNA系统发育树分析结果证实,该菌株为美人鱼发光杆菌杀鱼亚种(Photobacterium damselae subsp.piscicida).该菌对庆大霉素、青霉素、氟哌酸、氧氟沙星、氨苄青霉素等药物高度敏感,对红霉素、链霉素、卡那霉素、苯唑青霉素等药物具有抗性.     

条石鲷胚胎及仔稚鱼的发育

通过对条石鲷胚胎发育和仔鱼发育形态的观察,确定了条石鲷早期发育的进程。条石鲷的受精卵为圆球形、透明、浮性,卵径为0．90～0．95mill,油球1个,卵裂为盘状卵裂。在水温23～24℃,盐度26～28的条件下,受精后7h 10mirh胚胎发育至原肠期;受精后12h,眼囊形成;受精后20h 30min,心跳开始搏动;受精后26h 30min仔鱼孵出。初孵仔鱼全长1．78～2．03min,至5日龄全长3．25～3．30min时,卵黄囊完全消失;至8～15日龄,全长3．50～4．97mm时,鳔形成;至18．20日龄,全长7．10～7．31min时,尾鳍鳍条发育完整。     

ACE Inhibitory Peptide from Skin Collagen Hydrolysate of Takifugu bimaculatus as Potential for Protecting HUVECs Injury

Angiotensin-I-converting enzyme (ACE) is a crucial enzyme or receptor that catalyzes the generation of potent vasopressor angiotensin II (Ang II). ACE inhibitory peptides from fish showed effective ACE inhibitory activity. In this study, we reported an ACE inhibitory peptide from Takifugu bimaculatus (T. bimaculatus), which was obtained by molecular docking with acid-soluble collagen (ASC) hydrolysate of T. bimaculatus. The antihypertensive effects and potential mechanism were conducted using Ang-II-induced human umbilical vein endothelial cells (HUVECs) as a model. The results showed that FNLRMQ alleviated the viability and facilitated apoptosis of Ang-II-induced HUVECs. Further research suggested that FNLRMQ may protect Ang-II-induced endothelial injury by regulating Nrf2/HO-1 and PI3K/Akt/eNOS signaling pathways. This study, herein, reveals that collagen peptide FNLRMQ could be used as a potential candidate compound for antihypertensive treatment, and could provide scientific evidence for the high-value utilization of marine resources including T. bimaculatus.     

饲料蛋白能量比对红鳍东方鲀幼鱼的生长性能、饲料利用和体组成的影响

为探求红鳍东方鲀(Takifugu rubripes)幼鱼饲料的最适蛋白质能量比,以鱼粉和豆粕作为主要蛋白源,鱼油和豆油作为主要脂肪源,配制粗蛋白含量分别为36%、42%和48%,粗脂肪含量分别为8%、12%和16%,蛋能比为17.06~24.20 mg/kJ的9组饲料。投喂初始体重为14.95 g的红鳍东方鲀幼鱼56 d。结果显示,由双因素分析方法得出,饲料的蛋白水平、脂肪水平和蛋能比水平均可显著影响红鳍东方鲀的终末体重、特定生长率(SGR)和饲料效率(FE),且饲料蛋白为36%时,显著低于42%和48%组,但与饲料蛋白为42%和48%组之间相比无显著差异。在饲料脂肪为8%时,显著低于12%组,与饲料脂肪为12%与16%组之间相比无显著差异。此外,饲料的蛋白和脂肪水平对红鳍东方鲀生长和饲料利用的相关指标均无显著的交互作用(P>0.05)。由单因素方差分析得出,终末体重、FE、SGR均是Diet8(48/12)蛋白水平为48%、脂肪水平为12%处理组最高,显著高于Diet1(36/8)蛋白水平为36%和脂肪水平为8%处理组、Diet2(36/12)蛋白水平为36%和脂肪水平为12%处理组、Diet3(36/16)蛋白水平为36%和脂肪水平为16%处理组(P<0.05)与Diet4(42/8)蛋白水平为48%和脂肪水平为8%处理组、Diet5(42/12)蛋白水平为42%和脂肪水平为12%处理组、Diet6(42/16)蛋白水平为42%和脂肪水平为16%处理组、Diet7(48/8)蛋白水平为48%、脂肪水平为8%处理组之间相比无显著差异(P>0.05)。因此,根据研究结果综合考虑生长性能及蛋白质节约效应,红鳍东方鲀幼鱼的最适蛋白水平为42%,脂肪水平为12%,蛋能比为20.75 mg/kJ。     

水解单宁对暗纹东方鲀摄食偏好、消化代谢和抗氧化能力的效应

为了解菜粕替代鱼粉后单宁的作用,配制4组等氮等能饲料饲喂初重(39.84±3.09) g的暗纹东方鲀8周。以含43%鱼粉的实用基础饲料T0为对照;另设置3个不同的鱼粉用量组,用酪蛋白平衡蛋白质含量,并添加0.25%(T1)、0.75%(T2)和1.25%(T3)的水解单宁,使酪蛋白:水解单宁≈菜粕中蛋白含量:菜粕中单宁含量=16.8∶1。结果显示,随饲料中水解单宁添加量的增加,增重率呈上升趋势,饲料系数显著下降;肌肉粗脂肪含量T2组显著高于T0及T1组,对照组氨基酸水平显著高于其他组。肝脏抗氧化指标中,T0组过氧化氢酶(CAT)活力显著低于其他组,总抗氧化能力(T-AOC)则显著高于其他组,T3组丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性显著高于其他组。血清抗氧化指标中,3个单宁组MDA含量均显著高于对照组,过氧化氢酶(CAT)活力显著下降。随饲料中水解单宁添加量的增加,血清谷丙转氨酶(ALT)活性先升后降,与肝脏中趋势一致;T1组白蛋白(ALB)含量高于T0及T3组,显著高于T2组,T1组总蛋白(TP)含量高于T0组,显著高于T2及T3组。T1及T2组肠道淀粉酶活性显著高于T0组,胃和肠道中的蛋白酶活性随水解单宁添加量的增加显著上升,胃脂肪酶活性显著下降。T3组苦味受体T2R1表达量在舌尖和肠道均显著高于对照组;肝脏热休克蛋白HSP70表达量T0组显著高于T3;摄食偏好实验中,驯化前暗纹东方鲀摄食偏好随水解单宁添加量的增加显著下降,驯化8周后,4组实验鱼对T2饲料的偏好程度均显著高于T0,T0饲料高于T1和T3组。研究表明,添加1.25%及以下水解单宁对暗纹东方鲀生长无负面影响,一定程度上对饲料有节约作用;添加水解单宁会改变鱼体对蛋白质、脂肪和糖类的消化及蛋白质合成代谢,添加量在1.25%时会损伤抗氧化能力和抗应激能力;用含有水解单宁的饲料驯化暗纹东方鲀可显著提高其对水解单宁添加量为0.75%的饲料的偏好程度,并提高对苦味的接受能力。     

松江鲈肾组织细胞系生物学特性研究

在25℃、5%CO_2实验室条件下,于20%FBS-DMEM/F12培养基中培养松江鲈肾组织细胞系,以研究其生物学特性。第75代松江鲈肾组织细胞传代培养后,0～2 d处于潜伏期,2～4 d进入对数生长期,4～6 d进入稳定期,其群体倍增时间为55.2 h。经液氮冷冻保藏1月后,复苏细胞的存活率为(90.8±1.37)%。35代松江鲈肾组织细胞的染色体数为2n=40,核型公式为K(2n)=16m+10sm+14t。病毒敏感性试验表明,松江鲈肾组织细胞对鱼类诺达病毒不敏感,对鲤春病毒血症病毒敏感,测定病毒滴度为10^6.53/mL。镉离子敏感性试验显示,松江鲈肾细胞存活率随着氯化镉浓度的升高而降低,半致死浓度为(130±9.1)μmol/L,可用来作为镉离子检测的生物学指标。     

急性铜胁迫对暗纹东方鲀组织铜积累、氧化应激、消化酶、组织病变及脂代谢相关基因表达的影响

通过 96 h 急性毒性实验研究水体铜暴露对暗纹东方鲀(Takifugu fasciatus)生理生化及脂代谢相关基因表达的影响。实验设置对照组、0.1 mg/L 和 0.2 mg/L (96-h LC50) 3 个铜处理组。结果表明,铜在暗纹东方鲀肝、肌肉和全鱼中的积累量随着处理浓度的提高而提高。同等浓度处理下,铜的积累量为肝脏>全鱼>肌肉。随着铜处理浓度的提高,肝脏中丙二醛(MDA)含量及抗氧化酶活性[谷胱甘肽过氧化物酶(GSH-PX),总超氧化物歧化酶(SOD)和过氧化氢酶(CAT)]显著上升。急性铜暴露诱发肝脏血细胞沉积以及血窦扩张的症状。在鳃中,诱发上皮细胞增生,顶部棒状以及产生动脉瘤等症状。急性铜胁迫后,暗纹东方鲀肠道中的淀粉酶活性显著上升,但脂肪酶活性显著下降。在肝脏中,随着处理浓度的提高,淀粉酶、蛋白酶和脂肪酶活性显著降低。在 0.1 mg/L 处理组,脂肪合成相关基因(G6PD、6PGD、LPL、Fas 和 Acc)的表达量最高。但在 0.2 mg/L 处理组中,脂肪分解相关基因(HSL 和 CPT 1)的表达最高。急性铜胁迫后对转运因子 PPARα的影响不显著,但转运因子 PPAR γ的表达量显著上升。本实验表明铜对暗纹东方鲀生理生化指标及脂代谢相关基因的表达均产生显著影响,本研究为暗纹东方鲀养殖过程中铜的合理使用提供有益的指导价值,也为生产中更好地监控重金属污染提供一定的技术指标参考。     

Dietary eicosapentaenoic acid requirement of juvenile rock bream,Oplegnathus fasciatus

A feeding trial was conducted to evaluate the optimum dietary level of eicosapentaenoic acid (EPA, 20:5n‐3) based on growth and non‐specific immune responses in juvenile rock bream. A basal diet without EPA supplementation was used as a control, and six other diets were prepared by supplementing with 4, 8, 12, 16, 20 or 40 g of EPA per kg diet. The actual EPA concentrations of the diets were 0.5, 4.3, 8.5, 13.0, 16.8, 21.0 and 41.2 g of EPA per kg diet, and the diets were abbreviated as EPA_0.5, EPA_4.3, EPA_8.5, EPA_13.0, EPA_16.8, EPA_21.0 and EPA_41.2, respectively. Triplicate groups of fish averaging 1.06 ± 0.01 g (mean ± SD) were fed one of the seven experimental diets at the apparent satiation for 8 weeks. At the end of the feeding trial, weight gain, specific growth rate and feed efficiency of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets (p < .05). Superoxide dismutase activity of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3 and EPA_8.5 diets. Fish fed EPA_21.0 and EPA_41.2 diets showed significantly higher lysozyme activity than did fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets. The broken‐line analysis of weight gain indicated that the optimum dietary EPA level was 16.7 g/kg diet. These results suggested that the optimum dietary EPA level in juvenile rock bream could be greater than 16.7 g/kg diet but less than or equal to 16.8 g/kg diet based on the broken‐line analysis and the ANOVA test of weight gain.