饲料中添加甜菜碱对尼罗罗非鱼生长_肌肉组成和消化率的影响

甜菜碱是甜菜加工副产品中提取的甘氨酸甲基内酯,甜菜碱以前仅限于作为动物营养的甲基供体,参与氨基酸的合成和协同作用。作为人类医学上的恢复胃酸的药物及肝脏保护性治疗剂等［朱仲贤１９７９］,参与氨基酸的合成和协同作用。甜菜碱作为鱼类的饲料添加剂的主要作用是...     

尼罗罗非鱼肌肉蛋白质双向电泳体系的建立

为建立1种尼罗罗非鱼肌肉组织蛋白质的双向电泳体系,实验将尼罗罗非鱼肌肉组织蛋白质提取后,用双向电泳(2-DE)分离蛋白质,分别对蛋白质样品的制备方法、IPG胶条长度及p H范围、SDS-PAGE凝胶浓度及染色方法4个关键因素进行了探索和优化。结果显示,采用液氮研磨-丙酮沉淀法制备样品蛋白质,使用24 cm p H 4~7的IPG胶条进行第一向等电聚焦电泳,第二向SDS-PAGE电泳采用浓度为12.5%的凝胶进行,双向电泳后的凝胶采用硝酸银染色法进行染色处理,该条件下扫描所得尼罗罗非鱼肌肉蛋白质二维电泳图谱具有蛋白质分离程度好、斑点清晰、分辨率高及横纹少等优点。研究表明,技术体系适用于尼罗罗非鱼肌肉蛋白质的分离,可用于后续罗非鱼肌肉蛋白质组学研究。     

龙胆草提取物对吉富罗非鱼肌肉品质的影响

以龙胆草提取物作为添加剂,按照不同比例配制等氮等能饲料,对罗非鱼幼鱼进行分组饲养.通过11周养殖试验,采集鱼体肌肉组织,测定不同添加量组间肌肉常规及其氨基酸组成等指标,以研究龙胆草作为饲料添加剂对罗非鱼生长及肌肉品质的影响.结果表明,龙胆草提取物作为饲料添加剂能够显著提升吉富罗非鱼具有提高肌肉粗蛋白含量,增加必需氨基酸以及鲜味氨基酸含量的作用,其中以0.3％为最适添加量.     

盐度、碱度对尼罗罗非鱼生长和肌肉品质的影响

在水温(24±1)℃下,将初始体质量(50.00±4.12)g的尼罗罗非鱼放入150 cm×60 cm×40 cm循环可控水族缸内,每箱15尾,每组设置3个平行。将尼罗罗非鱼在盐度(12 g/L)、碱度(23.8 mmol/L NaHCO3)、盐碱(12 g/L和23.8 mmol/L NaHCO3)以及淡水(对照组)水体中分别饲养56 d,比较和测量各组鱼体生长性能指标、肌肉常规营养成分、结合氨基酸和游离氨基酸含量,研究盐度、碱度对罗非鱼生长性能和肌肉品质的影响。试验结果显示,与淡水组相比,改变水体盐度、碱度对鱼体质量增加率、特定生长率和饲料系数均无显著影响(P>0.05)。盐碱组鱼肌肉灰分含量升高,粗蛋白含量下降;粗蛋白含量显著低于其他组(P<0.05);各组水分和粗脂肪含量差异不显著(P>0.05)。各组必需氨基酸量与氨基酸总量比值和必需氨基酸与非必需氨基酸比值分别为40%~41%和66%~69%,各组间差异不显著(P>0.05);处理组中必需氨基酸指数为47.06~59.66,盐碱组>碱度组>盐度组,盐碱组显著高于盐度组和碱度组(P<0.05)。碱度组、盐碱组和盐度组鲜味氨基酸总量分别为淡水组的1.78倍、1.74倍和1.63倍,且碱度组和盐碱组显著高于盐度组(P<0.05);盐碱组甜味氨基酸和游离氨基酸总量显著高于盐度和碱度组(P<0.05)。水体盐度、碱度均可有效改善罗非鱼肌肉营养价值和呈味特征,而碱度对肌肉呈味的影响效果更加明显。     

氯化胆碱对尼罗罗非鱼生长影响的对比试验

<正> 胆碱(choline)是动物营养的必要成份,作为甲基的供体产生乙酰胆碱,在生理上起着各种重要的作用.1985年我们分别在室内外进行氯化胆碱对尼罗罗非鱼生长影响的对比试验。试验是以生长比速、饲料系数、脂肪蓄积率及蛋白质利用率为评定指标。三次试验结果表明,对照组比胆碱组稍好,但两者差异不显著(p>0.05)。可以认为在本试验条件下,饲料中胆碱含量已满足尼罗罗非鱼生理上的需要,没有添加氯化胆碱的必要。     

饲料中添加维生素对尼罗罗非鱼生长的影响

本试验是以酪蛋白、马铃薯淀粉等组成的精制饲料。在65天饲养试验中所得结果表明,饲料中添加混合维生素对尼罗罗非鱼的生长、食欲及饲料效率等都有显著影响,其平均增重率快30％左右,饲料消耗降低25％。饲料中不添加混合维生素,但与青饲料结合投喂也能起到与添加混合维生素相同作用。     

脆化专用饲料对罗非鱼生长和肌肉品质的影响

为考察脆化饲料对罗非鱼(Tilapia)生长和肌肉品质的影响,实验分别以配合饲料(对照组)、脆化饲料(试验组)喂养体质量为(115.30±23.69)g的罗非鱼99 d。结果显示:配合饲料组和脆化饲料组罗非鱼的增重率分别为185.25%、94.96%;饲料系数为1.87、2.94;肥满度为15.23、12.50;肌肉粗蛋白含量为19.40%、18.26%;粗脂肪含量为0.75%、1.32%;胶原蛋白为2.03、2.91;肌原纤维耐折力为33.93μm、64.84μm;滴水损失为2.59%、1.82%。与对照组相比,在生长方面,脆化饲料组罗非鱼增重率、肥满度显著降低,饲料系数显著增加(P<0.05);在肌肉品质方面,肌肉的粗蛋白含量和滴水损失显著降低,而粗脂肪含量、胶原蛋白含量和肌原纤维耐折力显著提高(P<0.05)。结果表明,脆化饲料在一定程度上改善了罗非鱼的肌肉品质。     

甘露寡糖对断奶仔猪肠道菌群的影响

选取28日龄断奶的三元[杜×(大×长)]杂交仔猪96头,平均体重为(7.46±0.38)kg,按体重和性别随机分为4个处理组,每个处理组3个重复,每个重复8头仔猪,公母各半。第一个处理组为空白组,第二、三、四个处理组在相同的基础日粮中分别添加0.1%、0.2%和0.4%的甘露寡糖,研究甘露寡糖对断奶仔猪肠道菌群的影响。结果表明:(1)各实验组肠道大肠杆菌的浓度极显著低于对照组;(2)第三组和第四组其结肠、盲肠和直肠中双歧杆菌增殖显著。第二组中只有结肠中双歧杆菌显著高于对照组,但盲肠和直肠中双歧杆菌增殖不明显;(3)各个处理组肠道中乳酸杆菌的浓度与对照组相比差异不显著。     

Effects of dietary protein level on spawning performance of Nile tilapia (Oreochromis niloticus) broodstock reared at different water salinities

Effects of dietary protein level and water salinity on spawning performance of Nile tilapia broodstock and growth of their larvae were studied. Four isocaloric (400 kcal/100 g) diets containing 25%, 30%, 35% and 40% crude protein were prepared. The diets were fed to broodfish (25.7 g) reared at three water salinities (0‰, 7‰ and 14‰) at a female/male ratio of 3:1, to satiation twice a day for 195 days. The size at first maturation increased with increasing dietary protein at all salinities. At 25% and 30% protein levels, broodstock reared at 0‰ reached their sexual maturity at bigger sizes than those reared at 7‰ and 14‰. At 0‰, spawning intervals were not significantly affected by dietary protein levels. At 7‰ and 14‰, spawning intervals significantly decreased with increasing dietary protein level. Spawning frequency and number of eggs per spawn were increased with increasing dietary protein level. The total number of spawnings per female and absolute fecundity were better in fish fed 40% protein in freshwater than at 7‰ and 14‰ salinity. The relationship of dietary protein and water salinity on egg size was significant, but showed irregular patterns. The chemical composition of broodstock muscles, eggs and fry were not significantly affected by dietary protein and water salinity, except for body water and crude protein of broodstock which were significantly affected; but showed irregular trends. At each water salinity, egg hatchability was linearly increased with increasing dietary protein level. Eggs produced from broodstock fed 25% protein at 7‰ and 14‰ needed more time for hatching and yolk-sac absorption and resulted in poorer larval weight than those reared in freshwater. Fry growth was improved with increasing protein level at all salinities. This result revealed that 40% dietary protein is required for optimum spawning performance of Nile tilapia reared at 0‰, 7‰ and 14‰ salinity. It also indicated that spawning performance and larval growth were better in freshwater than at 7‰ and 14‰.     

尼罗罗非鱼Foxp1a/b的cDNA克隆及表达模式研究

为探讨低等脊椎动物Foxp1与免疫细胞活化及机体雌激素水平的相关性,本研究首次分离克隆了尼罗罗非鱼 (Oreochromis niloticus) Foxp1的开放阅读框序列,并通过real-time PCR对其mRNA表达水平进行检测。结果显示,尼罗罗非鱼具有两种不同基因编码的Foxp1分子,分别命名为OnFoxp1a与OnFoxp1b;OnFoxp1a为1710 bp,由15个外显子编码569个氨基酸,OnFoxp1b为2040 bp,由16个外显子编码679个氨基酸;OnFoxp1a/b均含有Foxp亚家族的特征性结构,即锌指结构、亮氨酸拉链样结构和叉状螺旋结构;与OnFoxp1a相比,OnFoxp1b与高等脊椎动物Foxp1具有更近的亲缘关系。Real-time PCR检测结果显示,OnFoxp1a/b几乎在所有组织中均有表达,OnFoxp1a在精巢中有高水平表达, OnFoxp1b在心脏中有高水平表达,同时OnFoxp1b在免疫相关组织如鳃、脾脏、肾脏、肠等均有中等水平表达;淋巴细胞多克隆刺激剂PHA、PMA、LPS刺激尼罗罗非鱼外周血单个核细胞（PBMC）,结果显示,50 μg/mL PHA和50 ng/mL PMA分别刺激6、12、24 h均显著增强OnFoxp1b的表达 (p＜0.05),20 μg/mL LPS刺激后,OnFoxp1b的表达出现先降低后升高的趋势 (p＜0.05);而OnFoxp1a的表达除50 μg/mL PHA 刺激24 h后有所升高,其余均无显著变化;雌激素处理6月龄雄性尼罗罗非鱼48 h,OnFoxp1a/b在肠、肾脏中的表达显著上调 (p＜0.05),而脾脏中无显著变化 (p>0.05)。综上所述,低等脊椎动物硬骨鱼类两种不同基因编码的Foxp1a/b均为哺乳动物Foxp1的同源分子,但其序列、结构特征、表达模式迥异,提示其功能发生歧化;同时两者的表达与淋巴细胞活化及机体雌激素水平密切相关。     

Effects of dietary protein and energy levels on spawning performance of Nile tilapia (Oreochromis niloticus) broodstock in a recycling system

The effects of dietary protein and energy level on spawning performance of Nile tilapia broodstock and hatchability of their eggs were studied. Nine diets containing three protein (30, 35 and 40%) and three energy (14.6, 16.7 and 18.8 MJ GE/kg) levels were prepared. The diets were fed to duplicate groups of broodfish (average weight of 60.5 ± 4.6 g for males and 39.4 ± 3.1 g for females) at 2–3% of their body weight, twice a day, for 150 days. At low protein level (30%), time to first spawning was significantly longer than at 35 and 40% protein levels, and increased with increasing dietary energy (decreasing protein-to-energy (P/E) ratio). At 35 and 40% protein levels, time to first spawning was not affected by dietary energy. Inter-spawning intervals (ISI) showed irregular patterns in relation to dietary protein and energy, however, at 40% protein the females tended to spawn at shorter intervals, regardless of dietary energy. Spawning performances, including total number of spawnings per tank, number of spawnings per female, absolute fecundity and average number of eggs per spawn, were all significantly lower (P < 0.05) at 30% CP than at higher protein levels. At all protein levels, increasing dietary energy from 14.6 to 18.8 MJ GE/kg (and decreasing P:E ratio) resulted in a significant decrease in fish fecundity. Egg size was not significantly affected by dietary protein and energy. At all energy levels, egg hatchability increased with increasing dietary protein levels (P < 0.05). Eggs produced from broodstock fed 30 and 35% CP, exhibited significantly lower hatchability (P < 0.05) and needed less time for hatching and yolk-sac absorption, and resulted in lower larval length than those fed a 40% protein diet. However, increasing energy level at each dietary protein level did not significantly affect these parameters. These results revealed that the best spawning performance of Nile tilapia broodstock reared in a recycling system was achieved at 40% dietary protein and 16.7 MJ GE/kg, with a P/E ratio of 23.6 g/MJ.     

Effects of Cu-exchanged montmorillonite on growth performance, microbial ecology and intestinal morphology of Nile tilapia (Oreochromis niloticus)

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to 4 treatments, each of which had three replicates of 30 fish/tank and used to investigate the effects of Cu²⁺-exchanged montmorillonite (Cu-MMT) on growth performance, microbial ecology of the skin, gill and intestine, and intestinal morphology. The dietary treatments were: 1) basal diet, 2) basal diet + 1.5 g/kg MMT, 3) basal diet + 30 mg/kg copper as CuSO₄ (equivalent to the copper in the Cu-MMT treatment group), or 4) basal diet + 1.5 g/kg Cu-MMT. The results showed that supplementation with Cu-MMT significantly improved growth performance as compared to control and fish fed with Cu-MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu-MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with a tendency of Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with control. Measurements of villus and microvillus heights at different sites of the intestinal mucosa indicated that dietary addition of MMT or Cu-MMT improved intestinal mucosal morphology. However, the microbial ecology of the skin and gill was not affected by the addition of CuSO₄, MMT or Cu-MMT. Supplementation with CuSO₄ had no (P > 0.05) effect on the growth performance, microbial ecology and intestinal morphology. The results showed that Cu-MMT exhibited antibacterial activity in vivo and protected intestinal mucosa from invasion of pathogenic bacterium and toxins, resulting in a positive effect on the growth performance.     

低温对罗非鱼基因组DNA甲基化的影响

以经过连续多代抗寒选育获得的尼罗罗非鱼耐寒品系为实验材料,采用DNA甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)分析方法从全基因组水平上探讨了低温适应对罗非鱼DNA序列中CCGG位点的甲基化水平的影响。结果显示,选用的18对选择性扩增引物共检测到849个位点。耐寒品系检测到的位点数为411个,对照组为438个,其中发生甲基化的位点数分别为72和104个,总甲基化率分别为17.52%和23.74%;全甲基化位点分别为37个和65个,全甲基化率分别为9.00%和14.84%;半甲基化位点分别为35个和39个,半甲基化率分别为8.52%和8.90%。结果分析表明,尼罗罗非鱼耐寒品系的总甲基化水平、全甲基化水平和半甲基化水平较对照组均有一定程度的下降。与对照组相比,尼罗罗非鱼耐寒品系基因组DNA甲基化总体水平下降了6.22%,其中以全甲基化位点变异为主,其下降幅度比例明显,为5.84%。由此可见,经过连续多代的低温胁迫可导致尼罗罗非鱼DNA甲基化水平发生改变,发生了去甲基化反应,表现为基因组甲基化程度降低的特征,说明了DNA甲基化与罗非鱼抗寒性反应密切相关。     

Performance of plastic biofilter media with different configuration in a water recirculation system for the culture of Nile tilapia (Oreochromis niloticus)

Three kinds of locally available plastic biofilter media with different configurations (plastic rolls, PVC pipes and scrub pads) were evaluated for their efficiency in organic waste removal from the effluents of an intensive recirculating tilapia culture system. A set of three types of solid-removing filters consisting of screened sedimentation; upflow sand as well as plastic bead filtration accomplished the mechanical filtration. Values of critical metabolic wastes like total ammonia nitrogen (TAN) (0.92 ppm) and nitrite-nitrogen (NO₂-N) (0.22 ppm) were found to be well within the acceptable limits, while other water quality parameters in the culture water were also maintained within the normal range by the filtration system. Removal rates of 3.46 g TAN/m³ per day and 0.77 g NO₂-N/m³ per day, as well as TAN and NO₂-N removal efficiencies of 29.37 and 27.3% respectively, were established to be the best for the plastic-roll biofilter medium as compared to PVC-pipe and scrub-pad media. Percent removal of TAN and NO₂-N per pass of the biofilter (25.49 and 26.3% respectively) and the specific TAN and NO₂-N removal rates (43 and 9.6 mg/m² per day) of plastic rolls were also found to be superior to the other two biofilter media. Pieces of PVC pipes as biofilter medium is recommended to be used in the biofilters in view of their cheaper cost.     

Effect of gelatinization degree and inclusion level of dietary starch on the characteristics of digesta and faeces in Nile tilapia (Oreochromis niloticus (L.))

Higher faeces removal efficiency can help and reduce the negative impact of faecal waste on water quality and on the environment. Physicochemical properties of feed ingredients/nutrients may change the faeces characteristics and the ratio of removed to non-removed solids. The aim of this study was to assess what physicochemical properties of the diet (viscosity, fermentation and dry matter) influence faeces characteristics and faeces removal using Oreochromis niloticus (L.) as the test animal. Four experimental diets were formulated by the addition of two types of starch differing in gelatinization degree (gelatinized and native) and two levels (high and low) of starch to a basal diet according to a 2 × 2 factorial design. The diets were assigned to 16 tanks with 40 fish each, with four replicates for each diet. Replacement of native starch by gelatinized starch improved faeces removal rate, growth and digestibility (P < 0.01), but reduced fermentation (P < 0.05) at the end of the intestine. Addition of gelatinized starch did not change viscosity and dry matter of the digesta at the end of the intestine. A high level of starch in the diet also increased digestibility, growth and faeces removal percentage (P < 0.05). Fermentation and dry matter content at the end of the intestine were not influenced by a high starch diet, but viscosity was higher at the high level of starch inclusion. Volatile fatty acid levels in the stomach of Nile tilapia were high in the treatments with gelatinized starch. In conclusion, intestinal fermentation induced by native starch had a negative impact on faeces removal efficiency. A higher inclusion level of starch resulted in a higher viscosity of the digesta, leading to higher faeces removal efficiency.     

盐度胁迫对尼罗罗非鱼免疫相关指标的影响

为探究盐度胁迫对尼罗罗非鱼免疫的影响,对体质量(35.0±5.0) g的尼罗罗非鱼进行了急性和慢性的盐度胁迫实验,对免疫相关指标进行了检测和分析。在急性盐度胁迫中,设置0、5和15盐度组,分别在胁迫后6、12、24、48和96 h进行取样,检测血清SOD、CAT、GSH-Px和AKP的活性。在慢性实验中,设置0、10、20和30盐度组,胁迫8周后检测血清SOD、CAT、GSH-Px和AKP活性,并进行了无乳链球菌易感性实验。结果显示:①血清中SOD活性在急性盐度胁迫6、12和24 h时都有随盐度上升而上升的趋势,但在96 h时盐度15组酶活性显著低于盐度5组;在慢性盐度胁迫下,各组的酶活性呈现出随着盐度升高而显著性下降的趋势。②血清CAT活性在急性盐度胁迫下12和24 h时呈现出随着盐度升高而显著下降的趋势;在慢性胁迫下不存在显著性差异。③血清中GSH-Px活性在急性和慢性胁迫后,均呈现随着盐度升高而降低的趋势。④血清AKP活性在胁迫后6 h随盐度升高呈现出显著下降趋势;在慢性盐度胁迫下,盐度20组显著低于其他实验组。⑤尼罗罗非鱼对无乳链球菌易感性实验中,盐度10组的易感性和盐度0组之...     

Effect of dietary cyanobacteria on growth and accumulation of microcystins in Nile tilapia (Oreochromis niloticus)

A 12-week growth trial was conducted in a flow-through system to investigate the chronic toxic effect of dietary intake of cyanobacteria on growth, feed utilization and microcystins accumulation in Nile tilapia (Oreochromis niloticus L.) (initial body weight: 5.6 g). Six isonitrogenous and isocaloric diets were formulated to include different contents of cyanobacteria with the dietary microcystins increasing from 0 to 5460.06 ng/g diet. The results showed that dietary intake of cyanobacteria could increase the growth of tilapia while there are no impacts on feed conversion efficiency or mortality. Feeding rate was higher for the diets containing highest cyanobacteria. Microcystins were mostly accumulated in fish liver. The relationship between microcystins contents in muscle, liver, spleen and dietary intake could be described by quadratic equations.Microcystins content in the muscle of Nile tilapia in present study exceeded the upper limit of the tolerable daily intake (TDI) of microcystins suggested by the WHO (0.04 μg/kg body weight/d). It is suggested that Nile tilapia fed on toxic cyanobacteria is not suitable for human food.     

尼罗罗非鱼无乳链球菌荚膜多糖合成基因的表达及其与荚膜唾液酸含量、菌株致病性的关系

为了解尼罗罗非鱼无乳链球菌(GBS)荚膜多糖合成基因的表达及其与荚膜唾液酸含量、菌株致病性的关系,本实验克隆了尼罗罗非鱼荚膜多糖合成基因cps E、cps K和neu A,通过q RT-PCR方法分析了这3个基因在不同培养温度下表达水平的变化,同时通过比色法测定了不同培养温度下GBS荚膜唾液酸含量的变化,通过人工感染实验分析了不同水温条件下GBS对罗非鱼的致病性。结果显示,cps E、cps K和neu A编码的氨基酸序列均具有保守的与荚膜多糖合成相关的酶活性位点,Cps E、Neu A与已知鱼源GBS(Ia和Ib型)和人源GBS(Ia、Ib和II~IX型)相应序列的同源性均达到97%以上,而Cps K与鱼源、人源的序列同源性则分别为56%~100%和27%~100%。在不同培养温度下GBS cps K和neu A基因表达水平的变化与荚膜唾液酸含量的变化一致;在较高温度(28和34°C)下培养的GBS荚膜唾液酸含量及菌株攻毒后罗非鱼的死亡率均随温度的升高而递增,而在22°C下培养的GBS唾液酸含量最高,攻毒后罗非鱼的死亡率却最低。本研究结果表明,GBS Cps K和Neu A在GBS荚膜多糖的唾液酸化中起重要作用,较低温度下GBS荚膜唾液酸的高含量有助于其在宿主体内的潜伏,而较高水温条件下细菌的强致病性可能还与除荚膜唾液酸外的某些重要的毒力因子的表达有关。