Dietary lysine requirement of juvenile cobia (Rachycentron canadum)

An 8-week feeding trial was conducted to quantify the dietary lysine requirement of juvenile cobia with an initial average weight of 1.25 g reared in indoor flow-through and aerated aquaria. Six isonitrogenous and isoenergetic practical diets (44% CP and 16% lipid) containing six levels of dietary lysine ranging from 1.15 to 3.25% (dry weight) at about 0.4% increments, using fish meal and wheat gluten as sources of intact protein, supplemented with crystalline amino acids. Equal amino acid nitrogen was maintained by replacing lysine with nonessential amino acid mixture. Each diet was randomly assigned to three aquaria and was fed to apparent satiation by hand four times daily. The results indicated that there were significantly differences in growth performance and feed utilization among the treatments. Maximum weight gain, special growth rate and protein efficiency ratio occurred at 2.38% dietary lysine; but with the increase of dietary lysine from 2.38 to 3.25%, weight gain, special growth rate and protein efficiency ratio did not significantly increase. The hepatosomatic index, viscerosomatic index, condition factor, crude protein content in whole body and main composition in muscle were significantly affected by dietary lysine levels; however moisture, lipid and ash content in whole body were not significantly affected by the dietary lysine. There were significant differences in total serum protein, glucose and triacyglycerol concentrations in fish fed diets with different dietary lysine levels. Haematocrit and leukocyte count were significantly affected by dietary lysine level, but hemoglobin and red blood cell count were not significantly affected. Lysine concentration in serum was significantly increased with the increase of dietary lysine level from 1.15 to 2.38%. Broken-line analysis on the basis of special growth rate showed that the dietary lysine requirement of juvenile cobia was 2.33% of dry diet (5.30% dietary protein).     

军曹鱼的胚胎发育及仔稚鱼形态观察

为了探明军曹鱼早期发育阶段的特征和规律,本实验通过连续显微观察,记录分析其胚胎及仔稚鱼发育各时期的形态特征和发育特点。结果显示,军曹鱼受精卵呈圆球形,为浮性卵,卵径(1.245±0.065) mm,油球1个,油球直径(0.325±0.027) mm。在水温(27.0±0.5)℃、盐度29、pH值8.3的条件下,军曹鱼卵从受精至孵化出膜约需26 h 30 min,经历合子、卵裂、囊胚、原肠胚、神经胚、器官形成和孵化出膜7个阶段,共24个时期。初孵仔鱼平均全长(3.254±0.096) mm,仔鱼在孵化后第3天开口摄食,眼囊变黑,卵黄囊被消耗约80%,第5天卵黄囊消失,第7天油球耗尽,转化为外源性营养。孵化后第14天全长(19.933±1.118) mm,各鳍鳍条形成,进入稚鱼期;22日龄稚鱼全长(41.140±3.779) mm,尾柄部开始出现鳞片,46日龄的后期稚鱼全长达(116.667±5.916) mm,体表完全覆盖鳞片,除尾鳍形状外,体形与成鱼相似。     

军曹鱼仔稚鱼脊柱和附肢骨骼的发育特征

为探明军曹鱼仔稚鱼早期脊柱及附肢骨骼的发育特征,本研究采用软骨-硬骨双染色技术,分别利用阿利新兰、茜素红对软骨、硬骨以及1～33日龄军曹鱼仔稚鱼全骨骼进行染色,系统观察并描述其脊柱和附肢骨骼的发育特征。结果显示,军曹鱼脊柱开始发育的标志为7日龄仔鱼中脉弓和神经弓的出现;13日龄稚鱼椎骨、神经弓和脉弓开始骨化;背肋、腹肋分别于17、20日龄开始骨化;29日龄稚鱼脊柱骨化完成。附肢骨骼骨化起始顺序依次为胸鳍、尾鳍、腹鳍、背鳍和臀鳍。胸鳍匙骨于4日龄出现,肩胛骨孔于12日龄出现,同时上匙骨开始骨化;乌喙骨与肩胛骨于20日龄开始骨化;第1尾下骨于5日龄出现,15日龄稚鱼尾杆骨、侧尾下骨和尾鳍鳍条开始骨化,18日龄稚鱼尾下骨开始骨化;腹鳍支鳍骨于17日龄延伸至匙骨,同时腹鳍开始骨化;臀鳍和背鳍于17日龄由前向后开始骨化。研究表明,军曹鱼在13日龄进入稚鱼期,早期发育阶段的骨骼发育特征与其功能性适应密切相关。本研究结果对研究军曹鱼早期骨骼发育与功能适应、优化养殖条件有重要作用。     

Effect of temperature on growth and energy budget of juvenile cobia (Rachycentron canadum)

Growth and energy budget of juvenile cobia (initial body weight ∼ 22 g) at various temperatures (23, 27, 31 and 35 °C) were investigated in this study. Maximal ration level (RL_max, %/day) increased as temperature (T, °C) increased from 23 °C to 31 °C but decreased at 35 °C, described as a quadratic equation: RL_max = −0.023T² + 1.495T − 17.52. Faecal production (f, mg g^− 1 day^− 1) increased with increased temperature (T, °C), described as a power function: lnf = 0.738lnT − 0.806. As temperature increased, feed absorption efficiency in dry weight (FAE_d, %), protein (FAE_p, %) and energy (FAE_e, %) all increased first and then decreased, but the variation of feed absorption efficiency was small, with ranges of 89.59-91.08%, 92.91-94.71%, 93.92-95.32%, respectively. Specific growth rate in wet weight (SGR_w, %/day), dry weight (SGR_d, %/day), protein (SGR_p, %/day) and energy (SGR_e, %/day) showed a domed curve relative to temperature (T, °C), described as quadratic equations: SGR_w = − 0.068T² + 3.878T − 50.53, SGR_d = − 0.079T² + 4.536T −59.64, SGR_p = − 0.084T² + 4.783T − 63.08 and SGR_e = − 0.082T² + 4.654T − 60.99, and SGR_w, SGR_d, SGR_p and SGR_e maximized at 28.5 °C, 28.6 °C, 28.4 °C, 28.5 °C, respectively, as calculated from the regression equations. The relationships between feed conversion efficiency in wet weight (FCE_w, %), dry weight (FCE_d, %), protein (FCE_p, %), energy (FCE_e, %) and temperature (T, °C) also took on a domed curve described as quadratic equations: FCE_w = − 0.726T² + 39.71T − 473.8, FCE_d = − 0.276T² + 15.31T − 190.6, FCE_p = − 0.397T² + 22.05T − 277.9 and FCE_e = − 0.350T² + 19.39T − 239.9, and FCE_w, FCE_d, FCE_p and FCE_e maximized at 27.4 °C, 27.8 °C, 27.7 °C and 27.7 °C, respectively, as calculated from the regression equations. Energy budget of juvenile cobia fed satiation was 100C = 5F + 67(U + R) + 28G at water temperature 27 °C and 100C = 5F + 70(U + R) + 25G at water temperature 31 °C, where C is food energy, F is faeces energy, (U + R) is excretion energy and metabolism energy, and G is growth energy.     

温度和体质量对军曹鱼生长及氮收支的影响

为调查军曹鱼生长和氮收支与温度和体质量的关系,采用鱼类生物能量学的原理和方法,开展了不同温度(21、27和33 ℃)和体质量(10、20、50和100 g)对军曹鱼生长和氮收支的影响研究,并建立了生长——温度/体质量关系及不同温度和体质量条件下的氮收支方程。结果显示:(1)温度和体质量对军曹鱼生长影响显著,且两者之间存在交互作用。军曹鱼在实验的最高水温(33 ℃)及最小体质量(10 g)时生长最快,在实验的最低水温(21 ℃)及最大体质量(100 g)时生长最慢。(2)实验的体质量范围内,军曹鱼特定生长率均随水温升高呈显著增长趋势;在实验的温度范围内,军曹鱼特定生长率随体质量增加呈减速下降趋势,两者间的关系采用幂函数定量描述。多元回归分析表明,模型LnSGR=a+bLnW+cT+dT²+eTLnW能较好地拟合军曹鱼特定生长率、温度及体质量间的关系。(3)军曹鱼摄食氮、排粪氮、排泄氮和生长氮受温度和体质量影响显著,随温度升高而增加,随体质量增加趋于减少。(4)不同温度和体质量组军曹鱼氮收支方程存在差异,且温度对氮收支方程的影响大于体质量。对同一体质量组,水温27和33 ℃时生长氮占食物氮的比例明显高于水温21 ℃时,而水温27和33 ℃时代谢氮占食物氮的比例则明显低于水温21 ℃时;对同一温度组,军曹鱼氮收支方程变幅较小。(5)在所有氮收支方程中,摄食氮用于排泄的比例大于68%。研究表明,在实验的温度和体质量范围内,环境温度偏低不利于军曹鱼生长;排泄是军曹鱼氮支出的最主要途径;水温27~33 ℃时军曹鱼氮收支方程相对恒定。     

Dietary supplementation of mannan oligosaccharide on white sea bream (Diplodus sargus L.) larvae: effects on development,gut morphology and salinity tolerance

The influence of dietary mannan oligosaccharide (MOS) on the development, gut integrity and quality (in respect of stamina and survivability) of white sea bream Diplodus sargus L. larvae was investigated. White sea bream larvae were held under appropriate rearing conditions and fed Artemia, enriched by A1 DHA Selco^? with the addition or absence of MOS (Bio‐Mos®). The results indicated that larval growth performance and survivability were not affected by the MOS supplementation. Light microscopy revealed that MOS supplementation significantly improved the intestinal morphology by increasing the villi surface area by over 12%. Transmission electron microscopy revealed that MOS supplementation increased the microvilli length by 26% compared with the control. Salinity challenge experiments showed that MOS significantly increased larval stamina and survival in both 0 and 60 mg L^?1 salinity water by 13% and 22.9% respectively. These improvements in the larval quality at the early stages of fish development are important for the efficiency of intensive hatchery production.     

Effect of nitrite on larval development of giant river prawn Macrobrachium rosenbergii

The effects of ambient nitrite concentrations on larval development of giant river prawn Macrobrachium rosenbergii were evaluated. The trials were conducted in two phases: phase 1, larvae from stages I through VIII and phase 2, larvae from stage VIII until post-larvae. In both phases larvae were kept in water with nitrite (NO₂-N) concentrations of 0, 2, 4, 8 and 16 mg/L. Oxygen consumption was analyzed for larvae in stage II at nitrite concentrations of 0, 4, and 8 mg/L. Survival, weight gain, larval stage index and metamorphosis rate decreased linearly with increasing ambient nitrite concentration. However, there was no significant difference between larvae subjected to 0 and 2 mg/L NO₂-N. In phase 1, there was total mortality at 16 mg/L NO₂-N, while in phase 2 larval development stopped at stage X in this treatment. The oxygen consumption in stage II increased significantly at NO₂-N concentration from 0 to 4 mg/L, but there was no difference between 4 and 8 mg/L NO₂-N. In conclusion, increasing ambient nitrite up to 16 mg/L NO₂-N delays larval development, reduces larval growth rate and causes mortality, whereas no significant effect occurs for levels below 2 mg/L NO₂-N. However, the establishment of a general safe level of nitrite to M. rosenbergii hatchery may be difficult due to the great variability in larvae individual sensitivity.     

Effects of dietary phospholipid level in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>) larvae: growth,survival, plasma lipids and enzymes of lipid metabolism

A study was conducted to determine the effects of dietary phospholipid (PL) levels in cobia (Rachycentron canadum) larvae with regard to growth, survival, plasma lipids and enzymes of lipid metabolism. Fish with an average weight of 0.4 g were fed diets containing four levels of PL (0, 20, 40 and 80 g kg⁻¹dry matter: purity 97%) for 42 days. Final body weight (FBW), weight gain (WG) and survival ratio were highest in the 8% PL diet group and mortality was highest in PL-free diet group. We examined the activities of lipoprotein lipase (LPL) and hepatic lipase (HL) in liver, lecithin-cholesterolacyltransferase (LCAT) in plasma as well as plasma lipids and lipoprotein. LCAT activity showed a decrease of more than two-fold in PL-supplemented diet groups compared with the PL-free diet group. HL activity was highest in the 8% PL diet group and the other three groups showed no difference. LPL activity was significantly higher in the PL-supplemented diet groups than in the PL-free diet group. The dietary intervention significantly increased plasma phospholipids and total cholesterol (TC) levels, and the higher free cholesterol (FC) level contributed to the TC level. However, the fish fed PL exhibited a significantly decreased plasma triglyceride (TG) level. The lipoprotein fractions were also affected significantly by the PL. The PL-supplemented diet groups had significantly higher high-density lipoprotein (HDL) compared with the PL-free diet group, but showed a marked decrease in very low-density lipoprotein (VLDL). The results suggested that PL could modify plasma lipoprotein metabolism and lipid profile, and that the optimal dietary PL level may well exceed 80 g kg⁻¹ for cobia larvae according to growth and survival.     

吉富罗非鱼对饲料中维生素B1的需要量

采用维生素B1(VB1)含量为0.08(对照组)、0.57、1.13、2.09、4.11和8.09 mg/kg的6种纯化饲料,分别饲养初始体质量为(64.4±1.5)g的吉富罗非鱼12周,研究VB1对其生长性能、部分血清生化指标、肝脏VB1蓄积量及转酮醇酶基因表达量的影响,以确定其对饲料VB1的需要量.结果显示,随着饲料中VB1含量增加,吉富罗非鱼增重率先呈线性增加后趋于稳定,当饲料中VB1含量为1.13、2.09、4.11、8.09 mg/kg时增重率达最大.吉富罗非鱼肝脏VB1含量随着饲料VB1含量增加不断增大,当增加到2.09 mg/kg后趋于稳定.饲料中缺乏VB1显著提高血清丙酮酸含量(P＜0.05),但对全鱼水分、粗脂肪、粗蛋白、灰分无显著性影响(P＞0.05).饲料中添加VB1显著提高血清高密度脂蛋白胆固醇和肝脏转酮醇酶基因表达量(P＜0.05).饲料中VB1含量大于1.13 mg/kg各组的肝脏转酮醇酶活性显著高于VB1含量小于0.57 mg/kg组(P＜0.05).折线回归分析表明,吉富罗非鱼(64 ～325 g)获得最佳生长时对饲料VB1需要量为1.16 mg/kg;肝脏VB1蓄积量达到最大时,对VB1的需要量为2.06mg/kg.     

团头鲂对饲料中Zn的需求量

为了研究团头鲂对饲料中Zn的需要量,以平均体质量50 g的团头鲂为实验对象,采用半纯化饲料,以ZnSO4·H2O为锌源,设置Zn添加量分别为0、68、137、206、275 mg/kg(饲料中Zn总量为22.85、98.07、164.00、235.43、307.96 mg/kg)共5个Zn含量梯度,每个实验组设4个平行,在池塘网箱中养殖43 d。经过回归拟合分析,饲料中Zn添加量、饲料中Zn总量与团头鲂特定生长率、饲料系数、蛋白质沉积率、脂肪沉积率的关系,得到团头鲂对饲料中无机Zn补充量为155.86~161.25 mg/kg,对饲料总Zn需要量为184.85~190.39 mg/kg;在日均摄食量为5.12 g/100 g体质量下,团头鲂对饲料Zn日需求量为0.798~0.825 mg/100 g体质量,对饲料总Zn日需要量为0.946~0.974 mg/100 g体质量。饲料中Zn含量对团头鲂的成活率、肥满度、内脏指数、鳞片重/体质量、体质量/体长、脊椎骨重/体质量无显著影响(P0.05)。饲料中Zn含量对团头鲂的脊椎骨长/体长有显著影响(P0.05);饲料中Zn的补充,有利于团头鲂脊椎骨的生长。     

Early development of the shortfin silverside Chirostoma humboldtianum (Valenciennes, 1835) (Atheriniformes: Atherinopsidae)

The shortfin silverside Chirostoma humboldtianum has been considered for culture in Mexico, but success has been limited by a poor knowledge of its early development. First synthesis of the early development of the shortfin silverside is presented to determine conditions suitable for rearing. Brooder maturation was induced through photothermal cycles. C. humboldtianum ova were fertilized in vitro. The eggs were incubated in reconstituted water (160-180 mg/L CaCO₃) at 18 °C and 5 gm of NaCl per litre. During the hatching day, 300 shortfin silversides were stocked and followed up until metamorphosis in order to establish the timing of exogenous feeding, changes in food type, growth and development during critical periods for survival, according to the theory of saltatory ontogeny. Free embryos hatched 12 days after fertilization at 18 °C. First critical point for survival is the beginning of exogenous feeding. Free embryos started mixed feeding on day four of post-hatching (dph), point of no-return was presented towards the end of mixed feeding on 6 dph, larval period began at six (dph) when the anus is opened, and metamorphosis to juvenile was presented at 65 dph with a SL of 19.34 ± 2.28 mm, when scales and fins were well developed. Differences in growth between periods were detected: free embryos growth slower than larvae but mouth size depicted a larger growth rate in the former. Cephalic length and mouth size were negatively related to standard length in embryos and larvae. Mouth size was positively related to cephalic length in free embryos but negative in larvae. Results suggest that during the free embryo phase, growth priorities are directed to the development of apparatuses and systems; whereas, during the larval period, energy is directed to growth in length, mouth size and development of fins, which allows them to increase their swimming velocity, grants them a greater capacity to obtain exogenous food and, in consequence, increases fitness for survival.     

饲料中添加壳寡糖和霉菌毒素吸附剂对凡纳滨对虾肠道黏膜形态及菌群结构的影响

实验旨在研究在饲料中分别或联合添加壳寡糖(chitosan oligosaccharide,COS)和霉菌毒素吸附剂(mycotoxins adsorbent)对凡纳滨对虾(Litopenaeus vannamei)肠道黏膜形态及菌群结构的影响。在基础饲料(对照组)中分别添加250 mg/kg COS、2500 mg/kg霉菌毒素吸附剂、250 mg COS+2500 mg/kg霉菌毒素吸附剂(组号为C0、C0.25、M2.5、C0.25+M2.5),制成4组等氮等能饲料,投喂初重(0.23±0.02)g的凡纳滨对虾8周。结果显示:COS与霉菌毒素吸附剂联合使用有助于改善肠道形态学指标,C0.25+M2.5组绒毛高显著大于C0、C0.25组(P0.05),绒毛宽显著大于M2.5组(P0.05),肌层厚度显著大于其他各组(P0.05)。高通量测序结果表明各组有效操作分类单元(Operational Taxonomic Unit,OTUs)数目无显著性差异(P0.05);C0.25+M2.5组Observed species指数、Shannon指数、PD值显著低于M2.5组(P0.05),但显著高于C0.25组(P0.05),与C0组无显著性差异(P0.05);肠道菌群主要为变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes),C0.25+M2.5组变形菌门细菌含量最低且厚壁菌门细菌含量最高,各组拟杆菌门细菌均升高,C0组含量最低。在属水平上,弧菌属(Vibrio)、假交替单胞菌属(Pseudoalteromonas)、Spongiimonas、发光杆菌属(Photobacterium)以及Candidatus Bacilloplasma占主要优势,C0.25+M2.5组弧菌属、发光杆菌属细菌含量均有效减少;C0.25+M2.5组发光杆菌属细菌含量最低,假交替单胞菌属细菌含量最高。250 mg/kg COS和2500 mg/kg霉菌毒素吸附剂联合添加对菌群丰富度无显著影响,但优化了肠道形态学指标及肠道菌群结构,提升了肠道健康,效果优于单一添加组。     

盐度与pH对花鲈孵化、初孵仔鱼成活及早期幼鱼生长性能的影响

为探究我国北方海域花鲈繁育适宜盐度和pH处理的设定,及促进早期花鲈苗种推广,本研究针对不同盐度(0、15、20、25、30、35和40)与pH(5.5、6.5、7.5、8.5和9.5)条件下,北方人工繁殖花鲈受精卵的孵化率、畸形率及饥饿10 d后的存活情况进行了分析,并对花鲈初孵仔鱼进行不投饵耐饥饿实验,记录每日存活率(SR)及最终生存活力指数(SAI)。最后观测低盐0、养殖盐度30与高盐45对花鲈早期幼鱼生长性能的影响。结果显示,北方海域花鲈受精卵适宜孵化盐度为20~35,以盐度25组孵化率最高,适宜pH为6.5~7.5,以pH 6.5组孵化率最高。低盐15组中,具最低的孵化率及最高的畸形率,但在最终饥饿SR的统计中,低盐15组的SR及SAI明显高于盐度30与35组,推测低盐15接近花鲈体液等渗点,降低了渗透调节中的能量消耗,更利于存活。初孵仔鱼饥饿实验中,盐度20组SR下降相对平缓,在第8天仍有12.66%的SR(其他组已为0)。饥饿1 d时,各pH组SR均小于90%,以pH 6.5组SR最高,为89.11%。与盐度处理组相比,花鲈初孵仔鱼对pH变化较为敏感,其孵化率及最终SR显著低于盐度处理组。花鲈早期幼鱼盐度处理组中,低盐0与高盐45将对此规格花鲈幼鱼产生较大损伤,以高盐45抑制最显著,应为苗种盐度推广的上限,而花鲈生长盐度30与各处理盐度相比,较适合花鲈早期幼鱼的生长。研究结果将为北方海域花鲈繁育适宜盐度和pH处理的设定,及提高花鲈孵化率、育苗成活率与早期苗种推广提供基础资料。     

三疣梭子蟹低盐耐受性相关甲基化标记的开发及验证

为开发三疣梭子蟹低盐耐受性相关甲基化标记,采用甲基化敏感性高分辨率熔解曲线法(methylation sensitive high resolution melting curve, MS-HRM)进行了三疣梭子蟹低盐耐受性相关甲基化位点的筛选和应用。结果显示,从三疣梭子蟹转录组数据库中共开发了8个甲基化标记,其中6个标记与低盐度耐受性显著相关,在低盐度条件下表现出显著的去甲基化或甲基化,这些标记分别位于V-ATP酶(V-type proton ATPase)、CLC型氯离子通道、琥珀酸脱氢酶、NAD(P)转氢酶(NAD(P)、磷酸乙醇酸性磷酸酶、NADH还原酶基因,为三疣梭子蟹耐低盐新品种的分子标记辅助选育提供候选工具。研究表明,“宁象1号”耐低盐能力显著高于野生群体,且选育群体中位点Pt-M1、Pt-M2的去甲基化率达到100%,可能有利于低盐条件下保持体内Na⁺和Cl^-的平衡。     

光裸星虫胚胎和幼体发育的显微和超微结构

为探究光裸星虫(Sipunculus nudus)发育形态学特征,利用光镜和扫描电镜技术观察了光裸星虫胚胎和幼体发育的结构变化。结果表明:(1)光裸星虫胚胎及幼体发育经历卵裂、囊胚、原肠胚、担轮幼虫、海球幼体、匍匐幼体等主要阶段,卵裂、囊胚、原肠胚和担轮幼虫均在卵膜内发育;海球幼体出膜时,卵膜脱落,不发育为表皮。(2)水温为30~31.5℃条件下,卵子受精30 min后开始卵裂,2 h后形成原肠胚,4 h后发育为担轮幼虫,24 h后出膜成为海球幼体,8~9 d后附着变态为匍匐幼体。(3)光裸星虫为典型的螺旋式完全卵裂。担轮幼虫可做旋转式游动,胚体表面密布纤毛,其中,体前端具有一束较长的顶纤毛;纤毛通过卵膜孔中央伸出体外,分为普通纤毛和勺状纤毛两种类型。胚体形成眼点、肾管等器官。(4)海球幼体游动迅速,摄食浮游植物。口前纤毛环较短,头部具有眼点和神经节;口后纤毛环较长,纤毛类型也分为普通纤毛和勺状纤毛两种;随发育进行,体壁肌肉束不断增强,形成方格状纹理;胃部逐渐前移,肠道不断延长,发生螺旋,在躯干部前段形成肛门。(5)匍匐幼体在底质表面爬行,吞食砂砾;体表具有角质层,体壁肌肉发达;身体前端形成片状突起,将发育成触手;吻部具有规则排列的乳突;身体末端中央外凸形成球形尾器;收吻肌、腹神经索、固肠肌等器官或组织明显。研究结果可为光裸星虫生殖调控与人工繁育研究提供理论参考。     

金钱鱼抗缪勒氏管激素基因克隆及其在性腺发育不同时期mRNA表达水平的分析

为了解抗缪勒氏管激素基因(AMH)在金钱鱼性腺发育中的作用,本研究利用RACE技术克隆了AMH的cDNA序列全长,为2324 bp(GenBank登录号:KP718479),其开放阅读框为1631 bp,编码543个氨基酸。同源性分析显示金钱鱼AMH与花鲈相似性最高,为71.16%,与斑马鱼相似性仅为29.83%。系统进化树分析表明,该基因与鲈形目紧密聚为一支,与金钱鱼进化地位一致,说明AMH在进化中有一定保守性。氨基酸结构分析表明其1~28为信号肽序列,69~426为AMH-N区域,444~543为TGF-β结构区。实时荧光定量研究表明,金钱鱼成鱼中AMH基因在精巢中表达量显著高于其他组织,在肝和卵巢中也有表达。性腺不同发育时期分析表明,AMH基因在精巢发育Ⅰ、Ⅱ和Ⅲ期均维持高水平表达,IV期表达水平有所降低,H.E染色结果显示这一时期精巢发育逐渐成熟,推测该基因在精巢发育和精子产生过程中有重要作用。在卵巢中,AMH在Ⅰ、Ⅱ期卵巢发育初期表达量较低,在Ⅲ、Ⅳ期卵母细胞大生长期及卵黄积累期表达量升高,推测其在卵母细胞的发育和功能维持上发挥作用。提示AMH基因在金钱鱼精巢、卵巢发育过程中均发挥重要作用。     

吉富罗非鱼亮氨酸需求量研究

实验以初始体质量为(53.65±0.05)g的吉富罗非鱼幼鱼为研究对象,随机分为7个处理组(每个处理3个重复,每个重复14尾鱼),分别饲喂亮氨酸(Leu)水平为1.11%、1.51%、1.90%、2.29%、2.69%、3.08%和3.48%的7种等氮等能(粗蛋白32%)的半精制饲料56 d,旨在评价吉富罗非鱼对Leu的最适需求量。结果表明,以增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)为评价指标,通过二次回归分析可知,在投饲率为4%~6%时,罗非鱼饲料中适宜的Leu为2.28%~2.33%。随饲料Leu水平的增加,罗非鱼全鱼粗蛋白和粗脂肪、肌肉粗蛋白和粗脂肪均呈先升后降的趋势,而全鱼和肌肉水分差异不显著。罗非鱼胃蛋白酶、肠蛋白酶、肠脂肪酶、肠淀粉酶和Na+-K+-ATP酶的活性也随饲料Leu水平的升高呈先升后降趋势,除肠脂肪酶活性在2.69%水平组最高外,其他指标均在2.29%水平组达到最大值。同时,罗非鱼肠道表皮生长因子(EGF)、肠道表皮生长因子受体(EGFR)、血清超氧化物歧化酶(SOD)、血清溶菌酶(LZM)和血清碱性磷酸酶(AKP)也均在Leu为2.29%水平组达到最大值。研究表明,罗非鱼(50~200 g)饲料Leu的最适需求量为2.28%~2.33%(占饲料蛋白的7.11%~7.26%),该Leu水平能显著促进肠粘膜的发育,增强胃肠消化、吸收能力和机体非特异性免疫能力,从而促进罗非鱼的生长和饲料的转化。     

Morphological characterization of larval stages and first juvenile of the freshwater prawn Cryphiops caementarius (Molina, 1782) (Decapoda: Palaemonidae) under laboratory conditions

Research has been in progress for several years on various aspects of the biology and ecology of the freshwater prawn Cryphiops caementarius, an inhabitant of rivers in northern Chile. The commercial value of this prawn fomented the accomplishment of studies on its reproduction and development with the aim of producing juveniles under controlled conditions, to be followed by growout to commercial size in managed culture systems. The present study describes larval culture of this species from eggs of gravid females obtained in the field, from the first developmental stage (Zoea I) through the first juvenile stage. The larvae were cultured at 25 °C in UV sterilized water at variable salinities based on the requirements of the developmental stages. Larvae were fed with Nannochloris, Isochrysis and Artemia nauplii as required. This report describes in detail the 18 larval stages of this prawn, as well as its first juvenile form.     

糙海参胚胎和幼体发育的形态观察

为提高糙海参育苗技术,研究描述了糙海参从受精卵发育到稚参的形态变化,在显微镜下测定了受精卵、胚胎和幼体的大小,确定了糙海参胚胎发育的过程。结果表明,通过利用阴干、流水刺激法对成熟亲参进行人工催产,得到大量的受精卵,其受精率为90%以上。糙海参胚胎和幼体发育可分为受精卵、卵裂期、囊胚期、旋转囊胚期、原肠期、初耳状幼体、中耳状幼体、大耳状幼体、樽形幼体和稚参等阶段;在平均水温29℃,平均盐度34条件下,糙海参受精卵经3 h发育形成囊胚,4 h进入旋转囊胚期,5 h进入原肠期,19 h完成胚胎发育变态为耳状幼体;并经过7 d的生长与发育进入樽形幼体;第15天变态为稚参。观察发现,多精入卵现象则会导致胚胎发育不正常,最终使胚胎发育停止并死亡。在糙海参幼体发育过程中,大耳状幼体期幼虫臂的大小及其球状体的形成均可作为判断其幼体发育健康状况的重要指标:幼虫臂越大,球状体出现率越高,其幼虫的变态率和成活率也越高。对比发现,位于糙海参尾部的突出结构——尾突,为仿刺参和新西兰刺参所没有,这一结构差异同时导致了其骨片位置也不同;并且,糙海参胚胎和幼体与其他种类的海参在发育时间上存在一定差异。     

不同维生素E水平饲料对云纹石斑鱼幼鱼低盐胁迫前后抗氧化和渗透压调节功能的比较

维生素E(VE)在鱼类繁殖、生长代谢、抗氧化能力和免疫机能等方面有至关重要的作用。为探究饲料VE添加量对云纹石斑鱼低盐度胁迫下抗氧化和渗透压调节功能的作用,本实验设计0、20、40、80和160 mg/kg等5个VE水平饵料添加量,依次为A、B、C、D、E组,饲喂云纹石斑鱼幼鱼56 d。然后进行6 h低盐度(12)胁迫实验,测试皮质醇(COR)、葡萄糖(GLU)、乳酸(LD)等血清生化指标,超氧化物岐化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)和丙二醛(MDA)等抗氧化指标,以及鳃Na⁺/K⁺-ATP酶(NKA)、Ca²⁺-ATP酶活力等。结果显示,血清COR含量最高为盐度胁迫后A(32.74±1.53) ng/mL,最低为胁迫前E(19.06±3.88) ng/mL;GLU含量最高为盐度胁迫后A(16.46±0.99) mmol/L,最低为养殖前O(7.90±0.34) mmol/L;LD含量最高为低盐度胁迫前C(1.94±0.15) mmol/L,最低为胁迫后E组(1.30±0.06) mmol/L。说明VE的添加可以增强云纹石斑鱼抗应激的能力,且在47.524~134.566 mg/kg(B~D)的添加范围,效果较好。血清SOD、CAT、GSH-Px活力、T-AOC和MDA含量与投喂饲料中VE水平密切相关,SOD最高为胁迫前C(105.29±9.07) U/mL,最低为胁迫后A(11.23±2.30) U/mL;CAT最高为养殖前O(4.09±0.17) U/mL,最低为胁迫后A(0.35±0.10) U/mL;GSH-Px最高为胁迫前E(972.58±55.35) U,最低为胁迫后A(47.90±10.64) U;MDA最低为胁迫前D(33.48±2.34) nmol/mL,最高为胁迫后A(101.79±7.79) nmol/mL。饲料中添加91.378~178.924 mg/kg(C~E)水平VE对其抗氧化能力有增强作用。VE可增强NKA活力,当VE添加量为91.378 mg/kg(C)时,低盐度胁迫对NKA影响较小。而添加91.378~178.924 mg/kg(C~E)水平VE可增强云纹石斑鱼Ca²⁺-ATP酶活力。研究表明,饲料中添加91.378~134.566 mg/kg范围的维生素E,可以增强云纹石斑鱼的机体活力、抗应激及环境胁迫能力。