大口黑鲈开口摄食与转食人工配合饲料期消化系统发育特征

本研究利用组织切片和扫描电镜技术,观察和研究了2~30日龄大口黑鲈(Micropterus salmoides)仔鱼消化系统的发育过程及组织学变化。结果显示,在水温为(23±1)℃条件下,0~4日龄仔鱼消化道初步分化,为内源性营养期;4~6日龄仔鱼消化道逐渐分化形成食道、胃和肠,胃和肠黏膜褶形成,肝胰脏细胞团出现,仔鱼具备基本摄食能力,进入混合性营养期;10~16日仔鱼消化道和消化腺结构分明,胃、幽门盲囊、肠道紧密排列,肝脏和胰脏独立,进入外源性营养期,此阶段后可逐步转食人工配合饲料。20~30日仔鱼胃腺发达,胃和肠道出现次级黏膜褶,幽门盲囊黏膜褶显著增多、增长,肝脏逐渐出现脂肪积累区,胰脏可见酶原分泌颗粒,肝胰脏组织结构近似成鱼。扫描电镜显示,30日仔鱼胃部内表皮具有丰富的网状黏膜褶,胃小凹间分布着密集的分泌孔;幽门盲囊和肠道内表面结构相似,无固定形态的黏膜褶上布满黏液细胞和分泌孔。20日龄后仔鱼具备转食人工配合饲料的能力。此外,在仔鱼开口和转食人工配合饲料过程中,部分死亡个体的胃肠组织表现出腔体扩大或皱缩,内表皮无成型的黏膜褶或黏膜层脱落,胃和肠道组织损伤。本研究可为大口黑鲈仔鱼开口和转食人工配合饲料条件的优化提供组织学基础资料。     

大口黑鲈肌肉营养成分分析及营养评价

对三种不同来源(人工配合饲料饲养、冰鲜下杂鱼饲养和野生捕捞)的大口黑鲈成鱼的含肉率、肌肉中的水分、蛋白质、脂肪、灰分含量和氨基酸组成进行了分析,对其营养价值进行了综合评价,并与完全摄食人工配合饲料的中国花鲈进行了比较。结果表明:三种来源的大口黑鲈蛋白质含量在17.97%~20.15%,脂肪含量在0.81%~6.41%,灰分含量1.24%~1.41%,水分含量72.12%~79.98%,肌肉中17种氨基酸总量为14.19%~16.47%,其中必需氨基酸占氨基酸总量的44%以上,表明大口黑鲈肌肉蛋白是一种优质蛋白源。使用人工配合饲料饲养的大口黑鲈与使用冰鲜下杂鱼饲料饲养的大口黑鲈其肌肉营养成分以及氨基酸组成相差不大,但含肉率存在很大差异,人工配合饲料饲养、冰鲜下杂鱼饲养和野生捕捞的大口黑鲈含肉率分别为74.27%、64.86%和62.71%。而全部摄食人工配合饲料的中国花鲈含肉率为72.53%,氨基酸总量和必需氨基酸含量都高于三种来源的大口黑鲈。     

大口黑鲈肝细胞原代培养方法的建立

取材大口黑鲈肝脏组织,培养其原代细胞,比较研究了组织块法和酶消化法两种培养方法及不同的培养条件,以确定其最佳培养方法和条件,同时观察了长期培养过程中肝细胞的形态变化。结果表明:组织块方法不适于大口黑鲈肝细胞的培养,未见细胞从组织块中迁出。而胰蛋白酶消化法获得良好稳定的培养效果。胰蛋白酶浓度0.25%,消化时间20 min,分步收集肝细胞,经台盼蓝检测和血球计数板计数,平均活力>90%,每克肝重可获得3×106个分离肝细胞。在细胞活力和数量两方面达到最佳平衡。在含20%胎牛血清、10μg/mL胰岛素的M199/L15培养基中,于4%CO2,28℃培养箱中长期培养并传代。     

Lakeshore residential development and growth of largemouth bass (Micropterus salmoides): a cross‐lakes comparison

Gaeta JW, Guarascio MJ, Sass GG, Carpenter SR. Lakeshore residential development and growth of largemouth bass (Micropterus salmoides): a cross‐lakes comparison.
Ecology of Freshwater Fish 2011: 20: 92–101. © 2010 John Wiley & Sons A/S Abstract – Lakeshore residential development is associated with changes in littoral habitat, riparian habitat, and ecosystem function with potential impacts ramifying through aquatic food webs. Effects of these changes on economically important game fishes may vary with fish size. We investigated largemouth bass (Micropterus salmoides) size‐specific growth rates across 16 lakes spanning the range of lakeshore residential development in Wisconsin’s Northern Highland Lake District using a longitudinal multilevel model. Growth rates of small fish had a strong positive relationship with lakeshore residential development. The strength of the relationship decreased with length and became increasingly negative for fish longer than 210 mm. This pattern may be driven by a release from density‐dependent growth, shifts in available prey sources, reduced macrophyte cover, or angling‐induced selection pressures. Regardless of the mechanism, our results indicate, relative to undeveloped lakes, largemouth bass in highly developed lakes take 1.5 growing seasons longer to enter the fishery (356 mm).     

大口黑鲈肝脾肿大病病原研究

为了查明2009年10月广东省佛山地区养殖大口黑鲈(Micropterus salmoides)中暴发的传染性疾病的病原,对病鱼的肝脏、脾脏和腹隔膜进行切片电镜观察,发现细胞质中有大量病毒颗粒,切面为六角形,直径约145～150 nm,病毒为二十面体对称结构、无囊膜、似虹彩病毒的病毒粒子.用除菌的病鱼组织滤液感染健康大口黑鲈,被感染鱼死亡率达90%以上.根据已知虹彩病毒主要衣壳蛋白(MCP)基因序列设计特异引物,提取人工感染发病鱼的肝脏、脾脏、肾脏组织的DNA进行PCR扩增,将扩增片段进行序列测定与分析,结果表明该序列与已报道的鳜(Siniperca chuatsi)传染性脾肾坏死病毒(Infectious Spleen and Kidney Necrosis Virus,ISKNV)MCP基因同源性为98%.电镜观察和MCP基因测序分析结果显示,该病毒的分类地位为虹彩病毒科(Iridoviridae)细胞肿大病毒属(Megalocytivirus).     

大口黑鲈和中华绒螯蟹心脏组织结构及ATP酶含量的比较

研究比较了大口黑鲈(Micropterus salmoides)和中华绒螯蟹(Eriocheir sinensis H.Milne Edwards)心脏组织结构、肥大细胞和ATP酶含量.结果发现,大口黑鲈与中华绒螯蟹心脏组织结构明显不同.中华绒螯蟹心肌纤维排列更加松散,线粒体形态多样,嵴短而松散;大口黑鲈的心肌纤维有明显的明带和暗带,而中华绒螯蟹的仅见长度约为1.5 μm的暗带;大口黑鲈与中华绒螯蟹心肌组织间肥大细胞着色特性、形态及分布相似,中华绒螯蟹心脏组织中肥大细胞长径比大口黑鲈的略大,长约为20.6μm.透射电镜下观察,肥大细胞胞浆中充满高电子密度颗粒.在中华绒螯蟹心室中发现有与肥大细胞形态相似的白细胞.中华绒螯蟹心脏指数显著高于大口黑鲈,但大口黑鲈心肌组织ATP含量远高于中华绒螯蟹.结果表明大口黑鲈的心脏组织结构比中华绒螯蟹更为复杂,两种动物心肌间均发现有肥大细胞的分布,中华绒螯蟹单位心脏重量所含ATP酶的量明显比大口黑鲈低,这可能与它为开放式循环供能所需能量较低有关.     

饲料中添加酶解鸡浆对大口黑鲈肝脏抗氧化能力、肠道物理屏障和肠道菌群的影响

为评价酶解鸡浆在大口黑鲈健康养殖方面的功效,本实验用3.5%的鱼溶浆(SW)和酶解鸡浆(EC)分别等量替代基础饲料中的鱼粉,配制成3种等脂(EE 11.3%)实验饲料,在室内循环系统饲喂初始体质量为(9.25±0.13) g的大口黑鲈8周。结果显示,各处理组大口黑鲈增重率(WGR)、特定生长率(SGR)和摄食率(FR)均无显著差异。EC组肝脏总超氧化物歧化酶(T-SOD)活性、丙二醛(MDA)含量以及抗炎细胞因子(TGF-β)和促炎细胞因子(IL-8)基因表达量均显著优于对照组(基础饲料)和SW组。同其他处理组相比,添加酶解鸡浆后,肠道紧密连接蛋白(occludin、zo-1和claudin-1)基因相对表达量显著上调,而血清中二胺氧化酶(DAO)活性、D-乳酸(D-Lac)和脂多糖(LPS)含量显著降低。同时观察到酶解鸡浆会影响大口黑鲈肠道菌群的丰度(OTUs和Chao1),在属水平上增加潜在益生菌芽孢杆菌丰度,降低部分潜在有害菌如志贺氏菌属、不动杆菌属、弧菌属和支原体的丰度。进一步表型预测发现,添加酶解鸡浆会显著降低革兰氏阴性菌比例,增加革兰氏阳性菌比例。研究表明,饲料中添加酶解鸡...     

发酵豆粕、肉骨粉及其混合物替代鱼粉对大口黑鲈生长、血清生化指标和肠道组织学的影响

为探究2种蛋白源单一和混合替代鱼粉对大口黑鲈生长性能、血清生化指标和肠道组织学的影响,实验用发酵豆粕(FSM)、肉骨粉(MBM)和发酵豆粕+肉骨粉(FSM+MBM)混合物(1∶1)等蛋白替代基础饲料(FM,含鱼粉350 g/kg)中150 g/kg的鱼粉,使鱼粉含量降为200 g/kg,共配制成4种等氮等脂的实验饲料,记为FM (对照组)、FSM、MBM和FSM+MBM组,投喂初始体质量为(21.2±0.1) g的大口黑鲈8周。结果显示,与对照组相比,各替代组的增重率均无显著差异,但FSM组的增重率显著低于FSM+MBM组。此外,FSM组的饲料系数较对照组和MBM组显著升高。各组在全鱼成分(水分、粗蛋白、粗脂肪、粗灰分)和干物质消化率、蛋白质消化率以及蛋白质沉积率上无显著差异,而FSM组的蛋白质效率和脂肪沉积率显著低于对照组。此外,FSM组的血清谷草转氨酶显著增加,而血清总抗氧化能力以及超氧化物歧化酶、溶菌酶活性显著降低。在前肠形态学方面,各替代组的绒毛宽度均显著低于对照组,FSM组的绒毛高度、肌层厚度以及绒毛周长显著高于对照组。以上结果表明,在鱼粉含量为350 g/kg的饲料中,MB...     

Effects of different dietary lipids on growth,body composition and lipid metabolism‐related enzymes and genes in juvenile largemouth bass,Micropterus salmoides

This study investigates the effects of different lipids on growth, body composition and lipid metabolism of largemouth sea bass fish Micropterus salmoides. A total of 360 juvenile M. salmoides (mean ± SD mass = 33.83 ± 0.15 g) were randomly stocked into 12 tanks of 0.5 m³ volume for 8 weeks. Four replicates were made in each group, which were fed one of three diets containing fish oil (FO), soybean oil (SO) or lard oil (LO). The weight gain rate and specific growth rate did not differ among the groups (p > 0.05). Fish oil fish had the lowest condition factor (p < 0.05) and highest serum glucose content (p < 0.05). Crude lipid contents in the whole body and in the liver and muscle of FO fish were significantly lower than in the SO and LO groups (p < 0.05). The fatty acid composition of whole‐body lipids was closely correlated with that of the diet. The carnitine palmitoyltransferase 1 (cpt1) activity in the FO group was significantly higher than those in the SO and LO groups (p < 0.05). No significant differences in fatty acid synthase (fasn) activity were observed among the groups (p > 0.05). The Cpt1 and fasn gene expression levels in the FO group were significantly higher than those of the SO and LO groups (p < 0.05). The apolipoprotein B100 gene expression level was significantly higher in the SO group than in the FO group (p < 0.05). Fatty acid‐binding protein 1 gene expression levels in the FO and SO groups were not different (p > 0.05) but were both higher than that of the LO group (p < 0.05). The delta‐6 fatty acyl desaturase gene expression level in the LO group was significantly higher than that in the FO group (p < 0.05), but lower than that in the SO group (p < 0.05). It can be concluded that FO can be completely replaced by SO or LO in the M. salmoides diet, at least within the 8‐week culture period. Different types of dietary lipids significantly affect body condition and hepatic lipid metabolism in M. salmoides.     

大口黑鲈leptin A基因的组织表达及其对急性低氧胁迫的响应

瘦素(leptin)是一种重要的激素,参与调节动物的摄食、繁殖和能量消耗,其可以通过抑制食欲和促进能量代谢的方式维持机体能量稳态。大多数鱼类具有双重瘦素基因。在本研究中,利用PCR技术克隆了大口黑鲈leptin A基因的编码区,其开放阅读框(ORF)为486 bp,编码161个氨基酸蛋白。通过与其他物种进行同源性比对,发现鲈形目的 leptin基因的保守性较高,一致性高达91.46%,在大口黑鲈leptin A中几乎不存在基因特异性突变位点,而且大口黑鲈leptin A氨基酸序列与鳜同源性最高(92.59%),与花鲈(89.51%)、布氏鲳鲹(87.04%)、斜带石斑鱼(83.87%)的同源性次之。组织分布结果显示,leptin A基因在肝脏中的表达量最高,在心脏、头肾、脑、中肾、肠、脾脏、鳃、肌肉和性腺中的表达量均较低。此外,对大口黑鲈进行不同程度急性低氧胁迫[重度低氧(1.2±0.2)mg/L和中度低氧(3.5±0.2)mg/L],发现低氧胁迫初期时,严重低氧组和中度低氧组的leptin A的表达量都升高,显著高于对照组Leptin A的表达。这表明急性低氧能够诱导大口黑鲈leptin A在肝脏中的表达。综上所述,大口黑鲈leptin A基因与鳜leptin A基因的同源性最高,leptin A主要在大口黑鲈肝脏中显著表达,急性低氧胁迫能显著诱导leptin A的表达。