Dietary n‐3 highly unsaturated fatty acids affect the biological and serum biochemical parameters,tissue fatty acid profile,antioxidation status and expression of lipid‐metabolism‐related genes in grass carp,Ctenopharyngodon idellus

A 95‐day feeding trial was conducted to determine the effects of n‐3 highly unsaturated fatty acids (n‐3 HUFA) on the growth, antioxidation and lipid metabolism in grass carp (Ctenopharyngodon idellus). Two isonitrogenous and isolipidic diets were formulated with either lard oil (LO) or fish oil (FO) as the main lipid source. The results showed that the intraperitoneal fat (IPF) ratio was significantly lower (P < 0.05) in FO group. The concentration of n‐3 HUFA in muscle, hepatopancreas and IPF was significantly higher in FO group (P < 0.05). The serum low‐density lipoprotein (LDL) content was significantly lower (P < 0.05), and glucose (GLU) content was significantly higher (P < 0.05) in FO group. The serum total superoxide dismutase (T‐SOD) activity was significantly higher (P < 0.05) in FO group, consistent with the serum malondialdehyde (MDA) content. The gene expression of IPF fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), sterol regulatory element‐binding protein (SREBP‐1) and peroxisome proliferator‐activated receptor γ (PPARγ) was significantly lower (P < 0.05) and that of peroxisome proliferator‐activated receptor α (PPARα) was significantly higher (P < 0.05) in FO group compared with LO group. Similar trends were found in the hepatopancreas, except for PPARγ. It is suggested that n‐3 HUFA could inhibit lipid accumulation in grass carp by affecting the expression of lipid‐metabolism‐related genes.     

Effects of dietary macroalgae meal and lipid source on growth performance and body wall fatty acid composition of sea cucumber Apostichopus japonicus

A 70‐day experiment was conducted to examine the effects of different macroalgal meals and lipid sources on growth, body wall composition and fatty acid (FA) profile of sea cucumber Apostichopus japonicus. Two macroalgal meals including Sargassum muticum (SM) and Gracilaria lemaneiformis (GL) and two lipid sources including fish oil (FO) and vegetable oil (VO) were formulated into four diets, i.e., S. muticum and fish oil (SF), S. muticum and vegetable oil (SV), G. lemaneiformis and fish oil (GF) and G. lemaneiformis and vegetable oil (GV). The results showed that the specific growth rates (SGR) of A. japonicus fed diets containing SM were significantly higher than those fed diets containing GL. No significant differences in SGR between the FO‐based and VO‐based groups were observed. Similar results were observed in the body wall lipid content. Most body wall FAs changed to resemble the dietary FA proportions because of the dietary effect. Concentrations of 20:4n‐6 of the SF and GF groups were significantly lower than the SV and GV groups, while levels of 20:5n‐3 and 22:6n‐3 were significantly higher than the SV and GV groups. The n‐3/n‐6 polyunsaturated fatty acids (PUFA) ratios of the SF and GF groups were significantly higher than the SV and GV groups. Moreover, the SF group had significantly higher 20:5n‐3 and 22:6n‐3 contents and n‐3/n‐6 PUFAs ratio than the GF group. These findings reveal that the SF diet can show beneficial effects on both growth performance and body wall n‐3 PUFAs content of A. japonicus.     

Effects of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus

This study was conducted to evaluate the effects of different concentrations of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus. Diets were formulated to contain 40% crude protein from solvent-extracted menhaden fish meal and 0, 7, 14 or 21% lipid from menhaden fish oil. The basal diet, without supplemental fish oil, contained lipid at 0.4% of dry weight. The diets were fed to groups of 25 juvenile red drum initially averaging 7.3 ± 0.18 g fish^–1 in a recirculating culture system for 8 weeks and weight gain was recorded. After an additional 8 weeks, 16 fish from each treatment were sacrificed and the following measurements were recorded: hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, and liver -tocopherol, malondialdehyde (MDA) formation, and cytochrome P-4501A activity (measured as 7-ethoxyresorufin O-deethylase (EROD) activity). The activity of alanine and aspartate aminotransferases and concentrations of -tocopherol also were measured in plasma.Weight gain was significantly (p<0.05) affected by dietary lipid concentration, with values ranging from 361% of initial weight for fish fed the basal diet to 527% of initial weight for fish fed the diet containing 7% lipid. The HSI and IPF ratio values also were significantly affected by lipid with the lowest values recorded for fish fed the basal diet and the highest values observed in fish fed the diet containing 21% lipid. Increasing dietary lipid significantly increased oxidative stress as reflected in reduced -tocopherol in liver and plasma and increased MDA formation in the liver, although no overt pathological signs were observed. These findings suggest that lipid concentrations between 7 and 14%, when the diet contains 60 IU vitamin E kg^–1, are likely to limit oxidative stress and result in normal physiological responses of red drum.     

养殖大菱鲆肠道中大菱鲆弧菌的分离鉴定及药敏试验

2014年11月,大连地区某大菱鲆养殖场发生病害并伴有死亡,主要症状为吻部下颌明显出血,腹腔积水,肝脏充血,肠道出血有白便。从患病大菱鲆肠道中分离出1株可在硫代硫酸盐—柠檬酸盐—胆盐—蔗糖琼脂培养基上生长的菌株HZ-C1,人工回接感染试验证明其对健康大菱鲆具有较强致病性。通过形态学观察、16SrDNA序列同源性分析及生理生化试验确定菌株HZ-C1为大菱鲆弧菌。药敏检测结果显示,大菱鲆弧菌HZ-C1对氯霉素、丁胺卡那及头孢曲松等11种抗生素敏感,但对青霉素、四环素及诺氟沙星等15种抗生素耐药。本研究为大连地区养殖大菱鲆细菌性病害防治提供了一定病原学依据。     

Supplementation effect(s) of organic acids and/or lipid to plant protein‐based diets on juvenile yellowtail,Seriola quinqueradiata Temminck et Schlegel 1845, growth and,nitrogen and phosphorus excretion

A feeding experiment was conducted to investigate the effect of organic acids and/or lipid supplementation on growth, utilization and environmental loading of nitrogen (N) and phosphorus (P) in juvenile yellowtail fed fishmeal (FM) and plant protein (PP) diets. Six diets as FM (FM‐based), FM+P (FM with inorganic P), FM+L (FM with lipid), PP+CA (PP with citric acid), PP+L+CA (PP with lipid and citric acid) and PP+L+FA (PP with formic acid) were formulated. Yellowtails were fed each of the diets in duplicate groups; once a day, 6 days a week to near satiation at water temperature 19.0–25.0 °C for 16 weeks. Fishmeal with inorganic P gave the best growth while PP+L+FA the lowest. However, growth increased in PP+CA and PP+L+CA. Addition of lipid significantly increased N and P retention resulting in significant reduction in N and P excretion. Citric acid and FA supplementation to PP diets also increased retention of P; hence, its excretion was lowered. Thus, CA, FA and lipid in juvenile yellowtail diets can help to partially replace FM with PP sources and reduce inorganic P use to minimize environmental loading from aquafeeds.     

饲料中维生素A对青鱼幼鱼生长、血清生化指标和肝脏糖脂代谢酶活性及基因表达的影响

为研究饲料中维生素A(VA)对青鱼幼鱼生长、血清生化指标和肝脏糖脂代谢相关酶活性及基因表达的影响,实验选取360尾初始体质量为(6.10±0.10) g的青鱼幼鱼,随机分配至3个实验组中,每个实验组设置3个平行。采用单因素实验设计,以无维酪蛋白和明胶为蛋白源、菜籽油为脂肪源、糊精为糖源,同时添加矿物质混合物和维生素混合物(无VA添加)配制成3组实验饲料,分别以饲料1 (Diet1)、饲料2 (Diet2)和饲料3 (Diet3)表示。在饲料1、饲料2和饲料3中分别添加0、2 200和20 000 IU/kg VA醋酸酯(500 000IU/g),经高效液相色谱法(Agilent-1100, Agilent,美国)检测后实验饲料中VA的实际含量分别为178.2、2 058.9和18 436.2 IU/kg,养殖周期为8周。结果显示:饲料中VA缺乏会显著降低青鱼幼鱼的增重率(WGR)和特定生长率(SGR);VA缺乏会显著降低血清血糖(GLU)、甘油三酯(TG)和低密度脂蛋白(LDL)浓度,增加总胆固醇(TCH)浓度。饲料中添加2 058.9IU/kg VA能显著提高肝脏己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)活性,促进葡萄糖转运蛋白-2 (GLUT-2)、HK、葡萄糖激酶(GK)、PFK和葡萄糖-6-磷酸酶(G6Pase)基因表达。当饲料中VA含量为2 058.9 IU/kg时,对肝脏脂肪酸转运蛋白-1(FATP-1)基因表达无显著影响,但显著影响肉碱棕榈酰转移酶-1 (CPT-1)和肉碱棕榈酰转移酶-2 (CPT-2)基因表达。当饲料中VA缺乏时,CPT-1和CPT-2基因表达受到显著性抑制;当饲料中添加过量VA时,乙酰辅酶A羧化酶-2 (ACC-2)和脂蛋白酯酶(LPL)基因表达受到抑制;同时,VA过量组中肝脏过氧化物酶体增殖物激活受体-γ(PPAR-γ)基因表达显著下降。研究表明,在饲料中添加2 058.9 IU/kg VA可以促进青鱼幼鱼生长,提高肝脏对葡萄糖的转运能力,促进糖酵解和糖异生代谢平衡,同时促进脂肪酸合成和转运。     

饲料脂肪水平对俄罗斯鲟幼鱼生长、血液生化指标及抗氧化性能的影响

以鱼油、豆油、亚麻籽油等比例混合,配成脂肪含量分别为5.45%(L1)、7.00%(L2)、10.44%(L3)、14.37%(L4)、16.34%(L5)、18.91%(L6)及20.15%(L7)的等氮饲料,饲喂俄罗斯鲟(Acipenser gueldenstaedtii)幼鱼[(6.20±0.02)g]8周。实验结果表明,L3组的终末体质量、增重率最大,但与L2组间无显著差异(P0.05),L3组的蛋白质效率显著高于其它组(P0.05),且饲料系数最低。L6和L7组终末体质量、增重率和蛋白质效率显著低于L2和L3组(P0.05)。将幼鱼的增重率与饲料脂肪水平进行回归分析,得出俄罗斯鲟幼鱼在饲料脂肪水平为10.69%时生长最佳。随着饲料脂肪水平的升高,各组鱼的肝体比和体粗脂肪有上升趋势,其中肝体比在饲料脂肪含量大于14.37%时显著增大(P0.05)。血清中的甘油三酯、胆固醇含量也随脂肪水平升高而显著提高(P0.05),提示脂肪过高会影响俄罗斯鲟幼鱼的生长,降低蛋白质效率,导致脂肪在体内的过量积蓄。L7组鱼血清中高密度脂蛋白胆固醇含量最高,而L1组鱼的低密度脂蛋白胆固醇含量最高。肝脏脂蛋白酯酶活性随饲料脂肪水平的提高而升高,脂肪酸合成酶活性则随脂肪水平升高而降低,提示饲料脂肪水平的提高会促进鱼体脂肪的分解代谢。肝脏丙二醛含量和超氧化物歧化酶活性均随着饲料脂肪水平的提高显著升高(P0.05),表明脂肪含量过高会导致脂质过氧化,引发机体抗氧化酶活性的升高。综合俄罗斯鲟幼鱼的生长、血液指标及抗氧化性能,建议生产中将幼鱼的饲料脂肪水平控制在7.00%~10.69%。     

Plasma membrane of trout spermatozoa: I. Isolation and partial characterization

The plasma membrane from spermatozoa of rainbow trout was isolated by four techniques: sonication, hypotonic shock, mechanical homogenization after freeze-thawing, and nitrogen cavitation, in combination with continuous sucrose gradient centrifugation. Nitrogen cavitation (900 PSI, 20 min equilibration at 4°C) was the most effective technique.Following nitrogen cavitation, four bands were recovered in the sucrose gradient at densities 1.03, 1.05, 1.09 and 1.15 g/ml. Electron microscopy revealed membrane vesicles of various sizes in bands 1 to 3, while enzyme analysis revealed a 3.9 to 5.5-fold enrichment in 5'-nucleotidase and little contamination by lactate dehydrogenase (cytosol) and succinic dehydrogenase (mitochondria). Lipid analysis of bands 1 and 2 indicated a 6 to 7-fold enrichment in cholesterol and a cholesterol: phospholipid ratio of 0.59–0.70. Seven classes of phospholipids were present in bands 1–3 with no significant differences observed among bands. These data indicate that the vesicles (in bands 1 and 2) obtained after nitrogen cavitation are primarily plasma membranes. Membranes in band 3 appear to be slightly contaminated with nuclear membranes.Most of the plasma membrane proteins were acidic to neutral. The 2 main membrane proteins were 42 and 30 Kilodaltons.     

不同脂肪源饲料对中华绒螯蟹幼蟹生长、消化酶活力和脂肪酸组成的影响

选用初始体重为(2.15±0.10)g的中华绒螯蟹(Eriocheir sinensis)雄性幼蟹,随机分为5组(每组12只幼蟹),饲喂100%鱼油组(简称F1组)、100%豆油组(F2组)、100%亚麻油组(F3组)、50%鱼油+50%豆油组(F4组)、50%鱼油+50%亚麻油组(F5组)不同脂肪源配制的5种等氮等能饲料。实验蟹单个体养殖,实验周期为112 d。结果表明,F5组体重、增重率和特定生长率都显著高于其他饲料组(P0.05),各组的蜕壳间隔和肝胰腺指数没有显著差异(P0.05)。肝胰腺组织消化酶活力测定结果表明,F1组幼蟹肝胰腺的类胰蛋白酶活力显著高于其他各组(P0.05);F3组的胃蛋白酶活力显著高于其他各组(P0.05);F1组、F2组和F4组的脂肪酶活力显著高于其他两组(P0.05);各组的淀粉酶活力差异不显著(P0.05)。脂肪酸测定结果表明,肝胰腺和肌肉中的亚油酸(LOA)(C18:2n-6)、亚麻酸(LNA)(C18:3n-3)、EPA(C20:5n-3)和DHA(C22:6n-3)等主要脂肪酸含量与饲料脂肪酸组成呈正相关关系,F1组的高度不饱和脂肪酸(HUFA)含量显著高于其余各组(P0.05),F2组的LOA含量显著高于其余各组(P0.05),F3组的LNA含量显著高于其余各组(P0.05)。本研究结果表明,以50%的豆油或亚麻油替代鱼油能促进中华绒螯蟹幼蟹的生长,但可能使幼蟹的成活率降低。以豆油或亚麻油替代鱼油会影响幼蟹胰蛋白酶、胃蛋白酶和脂肪酶的活力和肝胰腺、肌肉脂肪酸组成。     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.