尼罗罗非鱼微卫星标记与主要生长性状的相关性分析

为了筛选到与尼罗罗非鱼生长相关的分子标记,并对这些标记进行准确性鉴定,运用65个微卫星标记对鹭业和番禺2个尼罗罗非鱼群体进行了PCR扩增,再利用SPSS软件一般线性模型(GLM)对这些微卫星位点与尼罗罗非鱼体重等主要生长性状进行了标记性状连锁关联分析。结果表明,鹭业群体中有8个微卫星标记(UNH130,UNH183,UNH911,GM558,UNH211,UNH176,UNH914和UNH974)与主要生长性状显著或极显著相关(P<0.05或P<0.01),其中有2个微卫星标记(UNH914和UNH974)与番禺群体的主要生长性状显著或极显著相关(P<0.05或P<0.01),1个微卫星标记(UNH176)只与体重显著相关(P<0.05)。通过对与生长性状相关标记的基因型和表型值进行多重比较,得到了对体重、体长和体高3种性状有利的基因型或等位基因。发现了对罗非鱼生长性状有显著效应的微卫星位点,为开展罗非鱼的分子标记辅助育种提供了有价值的遗传标记。     

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。     

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。     

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‰.     

尼罗罗非鱼人工驯养、选育群体遗传多样性及瓶颈效应

人工驯养和选育是家养动物适应性进化的主要动力之一,中国大陆尼罗罗非鱼引进群体经历了长期的人工驯养和选育,是研究鱼类在驯养、选育条件下适应性进化的良好材料。本实验以尼罗罗非鱼1个埃及土著群体为对照组,以中国大陆具有代表性的尼罗罗非鱼5个驯养群体和4个选育群体为实验组,采用12个多态性微卫星位点分析了驯养群体和选育群体的遗传多样性和瓶颈效应。结果显示,土著群体、驯养群体和选育群体平均每个位点的有效等位基因数(AE)分别为5.433、5.113~6.515和3.239~6.734,期望杂合度(HE)分别为0.812、0.796~0.859和0.657~0.858,多态信息含量(PIC)分别为0.768、0.753~0.819和0.601~0.818,近交系数(FIS)分别为0.323、0.166~0.342和0.249~0.314。LSDt检验结果显示,受人工选择的群体(驯养群体、选育群体)与土著群体间遗传多样性水平(AE和HE)无差异,3个驯养群体(EGY群体、WY群体和GD群体)的遗传多样性水平(HE)显著高于1个选育群体(XJF群体)。瓶颈效应分析显示,尼罗罗非鱼土著群体、驯养群体和选育群体在历史上都曾发生过群体缩小的现象。其中,土著群体、2个驯养群体(WY、EGY)、2个选育群体(JNM、XJF)在近期可能经历过遗传瓶颈,其他群体在新的突变和基因流的作用下,群体规模可能已恢复。有效群体大小分析显示,土著群体、驯养群体和选育群体的有效群体数量分别为177、29~117(平均值为57.4)和84~123(平均值为102.8)。本研究结果不仅为尼罗罗非鱼驯养群体的持续利用和选育群体的进一步遗传改良提供了有价值的信息,而且为鱼类在驯养和选育条件下群体遗传结构和种群动态研究提供了新的参考依据。     

不同蛋白水平下添加晶体氨基酸对罗非鱼生长和饲料利用的影响

实验主要研究不同蛋白水平下添加晶体氨基酸(CAA)对罗非鱼生长和饲料利用的影响。实验饲料为全植物蛋白实用饲料,通过调整豆粕含量,设置出4个蛋白梯度(蛋白含量:34%、32%、30%和28%),每个蛋白水平下设置不添加CAA组(34CP-28CP)和添加CAA组(34AA-28AA),后者通过添加赖氨酸、蛋氨酸和苏氨酸,使之在饲料中的水平分别达到1.43%、0.75%和1.05%,满足罗非鱼的必需氨基酸需要量。实验结果显示,34AA、30AA和28AA组的增重率和特定生长率(1074.75%、951.60%、815.12%和4.11%、3.92%、3.69%)显著高于相应的不添加组(869.85%、641.25%、523.75%和3.79%、3.33%、3.05%),32AA与32CP之间则没有显著差异(917.45%对882.27%、3.87%对3.81%)。从增重率的增长幅度来看,30%和28%蛋白水平下添加CAA的效果更为明显(48.4%、55.6%对23.6%、4%)。饲料效率、蛋白质效率、蛋白质保留率和能量保留率方面,34CP、34AA、32CP和32AA4组间没有显著差异;30CP和28CP组显著低...     

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。     

Bacterial diversity of tilapia (Oreochromis niloticus) cultured in brackish water in Saudi Arabia

The bacterial flora occurring in brackish pond water, sediment, gills and intestine of healthy tilapia cultured in Saudi Arabia were estimated both quantitatively and qualitatively, and the isolates were identified to genus or species level. Total viable count of bacteria ranged from 1.4±1.5×10³ to 8.6±2.7×10³ cfu ml⁻¹; 1.2±3.1×10⁶ to 7.3±1.1×10⁷ cfu g⁻¹; 8.7±1.9×10⁵ to 2.1±0.9×10⁶ cfu g⁻¹; and 2.8±2.4×10⁷ to 1.0±1.6×10⁸ cfu g⁻¹ in the pond water, sediment, gills and intestine of brackish water tilapia, respectively. In total, 19 bacterial species were identified. The bacteria were predominantly Gram-negative rods (87%). Pond water and sediment bacteria influenced the bacterial composition of gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. The predominant (prevalence >10%) bacterial species were Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus, Chryseomonas sp., Vibrio vulnificus, and Streptococcus sp. in all the populations with the exception of the sediment population where Streptococcus sp. was replaced by Shewanella putrefaciens. Vibrio spp. (58% of the total isolates) dominated the total bacterial population.     

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.     

甘露寡糖对高糖饲喂下尼罗罗非鱼肌肉品质的影响

甘露寡糖是一种新型饲料添加剂,它具有保护肠道健康和降血糖的功能。然而,甘露寡糖对于鱼类肌肉品质的影响并无系统性报道。为此,本实验选择均重为2g左右的健康尼罗罗非鱼幼鱼315尾,随机分成对照组（Control,糖含量为35%）、高糖处理组（HC,糖含量为45%）和高糖饲料中添加甘露寡糖处理组（HM,糖含量为45%,甘露寡糖添加量为0.5%）,每组3缸,每缸35尾,投喂10周后,测定其生长指标、肌肉营养成分和质构特性等相关指标。结果显示,与对照组相比,HC组尼罗罗非鱼肝体比,肌肉内聚性,肌纤维数量、肌苷酸含量、鲜味氨基酸占比、支链氨基酸/芳香族氨基酸比例、甘油三酯含量以及可促进肌肉生长的钙蛋白酶抑制蛋白基因CAST 相对表达量均显著升高;肌肉硬度、胶着性、咀嚼性、肌纤维直径、蒸煮损失,必需氨基酸占比以及在肌纤维的分化中起着重要作用的MyoG基因 相对表达量以及脂肪酸、甘油二酯和磷脂酰肌醇含量显著降低。与HC组相比,HM组尼罗罗非鱼的肌肉弹性、必需氨基酸占比,脂肪酸和甘油二酯含量以及CAST基因相对表达量显著增加;肌肉粘力、肌纤维数量、肌苷酸含量,磷脂酰乙醇胺和磷脂酰胆碱含量以及MyoG基因相对表达量均显著降低。与对照组相比,HM组尼罗罗非鱼增重率和肌肉内聚性,支链氨基酸/芳香族氨基酸比例,甘油三酯和脂肪酸含量显著增加;肝体比,肌肉硬度、咀嚼性以及磷脂含量均显著降低。上述结果表明,高糖饲喂可以使罗非鱼肌肉的保水能力增强,其肌肉更嫩更柔软紧密,提升肌肉鲜味,但会降低氨基酸营养价值。在添加甘露寡糖后,可以促进尼罗罗非鱼生长,提高产肉率和肌肉持水力,并且肌肉更加柔软紧密且富有弹性,提高氨基酸营养价值,但是会降低磷脂含量。     

尼罗罗非鱼无乳链球菌病的病理学研究

为了解尼罗罗非鱼感染无乳链球菌后各组织的病理变化,运用革兰氏染色和电镜负染技术对一株从自然发病的尼罗罗非鱼上分离的无乳链球菌进行形态观察,采用组织切片和超薄切片电镜技术对患病尼罗罗非鱼的肝脏、脾脏、肾脏、脑、心肌、骨骼肌、肠、鳃等8种组织进行病理学研究,探讨该病的致病机理。结果显示,革兰氏染色呈阳性,负染后透射电镜观察多数细菌呈链状排列;组织病理学变化主要是各内脏器官的广泛充血、水肿、变性和炎性细胞浸润,严重的细胞坏死;超微病理显示,大量球菌侵染脾脏等内脏组织,破坏细胞结构和各种细胞器;细胞界限模糊,细胞核畸形,线粒体肿大,嵴断裂,溶解;粗面内质网肿大、核糖体脱落;细胞质空泡化严重;心肌和骨骼肌纤维断裂、紊乱、肌节长短不一;肠微绒毛排列不整齐、长短不一;眼中有纤维性沉积。研究表明,无乳链球菌能造成尼罗罗非鱼全身性组织器官损害和炎症反应,尤其是肝脏、脾脏、肾脏和脑等重要器官功能障碍和衰竭,最后导致鱼体死亡。     

基于天然XY雌鱼培育YY超雄尼罗罗非鱼的新方法

前期报道了养殖群体存在天然XY雌鱼,但其能否用于培育YY超雄罗非鱼尚不清楚。本研究首先引入遗传性别受LG23染色体严格控制的CQ(重庆)尼罗罗非鱼群体和具有天然XY雌鱼的WC(湛江吴川)群体,将CQ群体XY雄鱼与WCXY雌鱼杂交,检验杂交F₁ YY超雄鱼是否可用于控制后代性别,并比较杂交F₁ XY和YY罗非鱼体质量、性腺指数、血清激素水平和性腺基因表达情况。研究发现,CQXY雄鱼和WCXY雌鱼交配,获得的F₁中有25%为YY超雄鱼,经鉴定为全雄且可育。将F₁ YY超雄鱼与WC XX雌鱼、WC XY雌鱼(母本)、杂交F₁ XX雌鱼和CQ XX雌鱼交配,后代几乎全雄,仅在与F₁ XX雌鱼交配的后代中有2尾雌鱼(雄性率98%)。在孵化后180 d,杂交F₁中XY和YY个体的体质量、性腺指数、血清激素水平差异不显著。基因表达分析发现,YY鱼精巢中AmhX/AmhY mRNA表达显著高于XY鱼,而Dmrt1和Cyp11b2 mRNA表达水平差异...     

Spontaneous diploidization of the maternal chromosome set in Nile tilapia (Oreochromis niloticus L.) eggs

Spontaneous diploidization of the maternal chromosome set (SDM) in Oreochromis niloticus is described here for the first time. The SDM phenomenon was observed in progeny of only one XY neofemale out of 11 such neofemales from which eggs had been fertilized with UV‐irradiated sperm: this treatment produced only the expected haploid embryos from the other females. SDM progeny were produced from three different batches of eggs from this female. No significant differences in survival at different stages of embryonic development were observed between the SDM and the diploid control group. The maternal inheritance of SDM progeny was verified using multilocus DNA fingerprinting. Only diploid karyotypes were observed in these fish and diploid control groups. In both the SDM and control groups, sex ratios were significantly skewed towards males (93.3% and 65.0% males respectively). The actual mechanism of SDM in eggs from this particular female remains unknown.     

尼罗罗非鱼鼠李糖凝集素 (RBL-1)在非特异性细胞防御病原菌感染中的功能

为了探究鼠李糖凝集素（Rhamnose-binding lectin, RBL）在硬骨鱼非特异性细胞防御病原菌感染中的作用,本研究以尼罗罗非鱼为研究模型,首先通过分离头肾单核/巨噬细胞进行体外菌应激实验,发现在罗非鱼两种重要的致病菌-无乳链球菌（Streptococcus agalactiae）和嗜水气单胞菌（Aeromonas hydrophila）应激后,尼罗罗非鱼L-鼠李糖凝集素1（OnRBL-1）的表达量显著上调。然后,通过荧光定量PCR（qRT-PCR）检测发现OnRBL-1重组蛋白能够调节病原菌诱导细胞炎症因子的表达,包括显著抑制菌诱导的IL-6、IL-8和TNF-α的表达,和促进IL-10和TGF-β的表达。此外,通过流式细胞术检测证实OnRBL-1具有促进单核/巨噬细胞的吞噬作用,同时增强呼吸爆发水平和上调活性氧的释放。以上研究结果表明,OnRBL-1在罗非鱼单核/巨噬细胞非特异性细胞防御中发挥重要的调节作用。本研究为探讨RBL-1在硬骨鱼宿主防御病原菌感染中的功能提供了参考,并有助于完善和丰富硬骨鱼类RBL的功能和在抗菌免疫应答中的基础理论体系,具有重要的科学意义。     

Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response

The aim of this study was to analyze the effect of a probiotic bacterium, Enterococcus faecium ZJ4 on growth performances and immune responses of tilapia (Oreochromis niloticus). The tilapias were treated with E. faecium ZJ4 at a final concentration of 1 × 10⁷ cfu ml^− 1 in aquaria water every 4 days. Six aquaria with three replicates for treated and controls were used. After 40 days, the tilapias supplemented with the probiotic showed significantly better final weight and daily weight gain (DWG) than those fed the basal diet (Control) (P < 0.05). There was no remarkable difference (P > 0.05) in the total serum protein, albumin content, globulin concentration and A/G ratio between the treated and control tilapias. The result of lysozyme activity assay was similar to these biochemical indexes. However, the complement component 3 content, myeloperoxidase (MPO) activity and the respiratory burst activity of blood phagocytes were higher (P < 0.05) in E. faecium treated tilapias (trial 1) than the controls.     

三种生态环境下尼罗罗非鱼免疫因子的比较

随机选取海水池塘(盐度28)和淡水池塘养殖及水库天然增殖的3种不同生态环境下、体质量为200~500g的尼罗罗非鱼为研究材料,采用生物化学方法测定3种生态环境尼罗罗非鱼血清中碱性磷酸酶、酸性磷酸酶、过氧化物酶、谷胱甘肽还原酶的活性和蛋白质量浓度。结果显示,3种生态环境中罗非鱼血清中碱性磷酸酶和酸性磷酸酶活力差异不显著(P0.05),其余各组间罗非鱼血清中过氧化物酶和谷胱甘肽还原酶的活性及血清蛋白质量浓度差异显著(P0.05),酶活力和蛋白质量浓度依次为:水库海水池塘淡水池塘。试验结果表明,水库天然增殖的尼罗罗非鱼具有较高的磷酸酶活性。     

水体盐度与饲料脂肪含量对尼罗罗非鱼生长、营养组成和肉质的影响

为研究水体盐度和饲料脂肪含量对尼罗罗非鱼生长、营养组成和肉质的影响,本实验设置0、8和16共3个盐度梯度,每个梯度分别投喂中脂(6%)和高脂(12%)饲料,投喂初始体质量为(5.0±0.2) g的尼罗罗非鱼8周,并测定生长性能、血清生化指标、肌肉营养成分和肉质相关指标。结果显示,在中脂饲料投喂下,与对照组(盐度为0)和高盐组(盐度为16)相比,中盐组(盐度为8)的尼罗罗非鱼具有最大终末体质量、躯壳比、脂体比、肌肉蛋白质含量、肌肉氨基酸和乳酸含量和总产肉率,但其饲料系数、脏体比、肝体比和肌肉p H值显著降低;而中脂高盐组的尼罗罗非鱼其饲料系数、肥满度、脏体比、肝体比、全鱼总脂、肝脏甘油三酯、血糖、血清乳酸和血清甘油三酯含量、血清谷草转氨酶活性较高,但全鱼和肌肉水分、全鱼灰分、产肉率和肌肉离心失水率降低。在高脂饲料投喂下,随着盐度的上升,其终末体质量、成活率和脂体比降低,但饲料系数、脏体比、肝体比和肥满度增大。其中,高脂中盐组尼罗罗非鱼的肌肉蛋白以及氨基酸含量显著降低,而高脂高盐组的全鱼灰分、肝脏甘油三酯、血糖、谷草转氨酶活性、肌肉总脂、肌肉甘油三酯和磷脂含量显著提高,但全鱼总脂、肝糖原、...     

尼罗罗非鱼淀粉酶性质的初步研究

研究了尼罗罗非鱼(Oreochromis niloticus)肝脏淀粉酶的基本性质及金属离子对该鱼淀粉酶活性的影响。结果表明:罗非鱼淀粉酶活力的最适pH是6.5,最适底物浓度是2%。研究金属离子对该鱼淀粉酶活力影响,一价金属离子K+、Li+、Na+对酶活力影响较小;二价金属离子Cu2+对酶活力具有抑制作用,Zn2+对酶活力无明显影响;三价金属离子Al3+对酶活力具有抑制作用,但效果不是很强烈,Fe3+对酶活力具有明显的激活作用;碱土金属Mg2+、Ca2+对淀粉酶活性有激活作用,而Ba2+对酶具有抑制作用;重金属离子Cd2+、Pb2+有明显的抑制作用。     

广东地区吉富罗非鱼无乳链球菌病的流行情况与耐药性

为了调查广东省惠州、肇庆、珠海、湛江4个吉富罗非鱼主养区链球菌病的流行情况和耐药性,并进一步分析β-内酰胺酶基因与青霉素类药物的耐药性关系。本实验通过传统的方法对菌株进行分离纯化,扩增特异性基因cfb以及16s r DNA对各菌株进行鉴定;采用k-b法测定分离菌株的药物敏感性;通过PCR检测β-内酰胺酶类基因在分离菌株中的分布情况,并用Statistic6.0统计分析各β-内酰胺酶基因与青霉素类药物的耐药关系。实验结果表明,吉富罗非鱼无乳链球菌的阳性率从高到低的顺序为惠州(46.46%)湛江(43.24%)肇庆(17.30%)珠海(4.17%);药敏结果显示各地区无乳链球菌分离株对青霉素(耐药率为94.29%)和磺胺二甲基嘧啶(耐药率为86.40%)普遍耐药,对恩诺沙星最为敏感(耐药率为3.99%);β-内酰胺酶基因分布与细菌耐药性统计结果显示,无乳链球菌基因组中的9个β-内酰胺酶基因在各分离菌株中的分布呈高度多态性,其中SAG0658基因与氨苄青霉素抗性显著相关,提示SAG0658基因在无乳链球菌耐氨苄青霉素过程中发挥主要作用;此外,9个β-内酰胺酶基因与青霉素抗性无相关性,说明其在菌株对青霉素耐药过程中并未发挥明显作用,提示分离菌株对青霉素的耐药性可能依赖其他途径。     

尼罗罗非鱼铁调素调节蛋白在宿主抵御病原菌侵染和调节铁稳态中的功能

为了研究铁调素调节蛋白(hemojuvelin,HJV)在硬骨鱼中抵御病原菌感染和维持自身铁稳态过程中的作用,实验扩增了尼罗罗非鱼铁调素调节蛋白基因(Onhjv)的开放阅读框(ORF),分析其在健康尼罗罗非鱼各组织中的分布模式及在抵御病原菌感染和调节铁稳态中的相关作用。结果显示,Onhjv的ORF全长由1 248个碱基组成,编码415个氨基酸,在不同物种之间具有一定的保守性。实时荧光定量PCR(qRT-PCR)结果显示,Onhjv在尼罗罗非鱼各组织中广泛分布,并在肝脏中的表达量最高。在无乳链球菌或嗜水气单胞菌感染后,Onhjv在肝脏、脾脏、肠和鳃中的表达量均显著上调。体外头肾单核/巨噬细胞和肝细胞中Onhjv表达量在受到这2种病原菌应激下也显著上调。此外,在1和10μmol/L FeCl_3溶液刺激后,Onhjv表达量在肝脏、脾脏、肠和鳃等组织,以及头肾单核/巨噬细胞和肝细胞中的表达量也呈显著上调。受重组罗非鱼IL-6蛋白[(r)OnIL-6]刺激后,头肾单核/巨噬细胞中Onhjv表达量显著上调,表明炎症因子可以促进Onhjv表达。研究表明,尼罗罗非鱼铁调素调节蛋白在宿主抵御病原菌感染和维持铁稳态的过程中发挥作用。本实验为探究HJV在硬骨鱼中的生物学功能提供了参考,同时为进一步研究铁代谢在宿主防御病原菌感染过程中的重要作用提供理论指导。