碳水化合物种类和水平对大黄鱼生长性能、血清生化指标、肝脏糖代谢相关酶活性及肝糖原含量的影响

本试验旨在研究碳水化合物种类和水平对大黄鱼生长性能、全鱼和肌肉常规成分、血清生化指标、肝脏糖代谢相关酶活性及肝糖原含量的影响。采用2×3双因素试验设计,选取葡萄糖和小麦淀粉2种碳水化合物,分别设0、15%、30%3个碳水化合物水平,共配制5种等氮等脂试验饲料。每种饲料饲喂3个重复,每个重复放养初始体重为(8.53±0.07)g的大黄鱼50尾,养殖试验持续8周。结果表明:饲料碳水化合物种类和水平及其交互作用对大黄鱼的终末体重、增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)有显著影响(P0.05)。饲料葡萄糖水平由0增加到30%时,大黄鱼的终末体重、WGR和SGR显著降低(P0.05);饲料小麦淀粉水平由0增加到15%时,大黄鱼的终末体重、WGR和SGR显著升高(P0.05),饲料小麦淀粉水平由15%增加到30%时,大黄鱼的终末体重、WGR和SGR显著降低(P0.05)。FCR随饲料葡萄糖水平的升高显著升高(P0.05);FCR在饲料小麦淀粉水平由0增加到15%时显著降低(P0.05),由15%增加到30%时显著升高(P0.05)。15%或30%水平下,小麦淀粉组大黄鱼的终末体重、WGR和SGR均显著高于葡萄糖组(P0.05),饲料系数显著低于葡萄糖组(P0.05)。饲料碳水化合物种类和水平的交互作用对大黄鱼血清总蛋白(TP)、总胆固醇(TC)、甘油三酯(TG)、葡萄糖含量有显著影响(P0.05)。血清葡萄糖含量随饲料葡萄糖的水平升高先降低后升高,各组间差异显著(P0.05);血清葡萄糖含量随饲料小麦淀粉水平的升高逐渐降低,且30%小麦淀粉组显著低于0小麦淀粉组(P0.05)。饲料碳水化合物种类和水平的交互作用对大黄鱼肝脏葡萄糖激酶(GK)、6-磷酸果糖激酶(PFK)、葡萄糖-6-磷酸酶(G6Pase)活性及肝糖原含量有显著影响(P0.05)。肝糖原含量随饲料葡萄糖的水平升高持续升高,而随饲料小麦淀粉水平的升高先升高后降低,且15%和30%水平下葡萄糖组均显著高于小麦淀粉组(P0.05)。由此得出,与葡萄糖相比,摄食含小麦淀粉饲料的大黄鱼能够通过调节糖代谢相关酶活性及肝糖原含量来维持血糖浓度的相对恒定,且饲料中添加15%小麦淀粉时能促进大黄鱼生长。     

饲料中不同淀粉源及水平对大口黑鲈肝脏组织学的影响

为了评估饲料中不同淀粉源及水平对大口黑鲈肝脏组织学的影响,采用4种淀粉(蜡质玉米淀粉、高直链玉米淀粉、小麦淀粉和木薯淀粉)各按两水平(5%和10%)配制成8种等氮和等能的饲料,投喂初始体质量为(23.45±0.08)g的大口黑鲈,经45 d的饲养后,采集肝脏样本进行肝脏组织的生化测定和切片观察。结果显示,摄食全部5%淀粉饲料和10%高直链玉米淀粉饲料的实验鱼的肝脏颜色呈正常的鲜红色;而摄食其他10%淀粉饲料的实验鱼肝脏均显现出发白现象,同时,肝细胞体积、肝体指数和肝糖原含量均显著高于5%淀粉饲料组和10%高直链玉米淀粉饲料组,肝细胞的细胞核移至细胞边位,出现空泡化现象。在5%和10%的淀粉水平上,高直链玉米淀粉饲料组的肝体指数、肝糖原含量和肝细胞体积均显著小于其他淀粉源的饲料组,肝细胞也没有出现细胞肿胀现象。本研究得出,饲料的碳水化合物来源与水平均对大口黑鲈肝脏组织的肝糖原含量和组织结构产生显著的影响;高直链玉米淀粉优于小麦淀粉、蜡质玉米淀粉和木薯淀粉,5%的饲料淀粉水平优于10%的水平。     

基于崩解时限的牡蛎糖原片剂配方组合优化研究

为了确定牡蛎糖原片剂的制备工艺,以崩解时限为重要指标,利用单因素结合正交试验法筛选优化牡蛎糖原片剂配方组合。研究结果表明,以牡蛎糖原冻干粉添加量15%、微晶纤维素15%、麦芽糊精5%、羧甲基淀粉钠5%、硬脂酸镁0.02%、玉米淀粉59.98%为片剂处方,优选出的处方工艺简单可行,制备的片剂质量符合中国药典(2015版)的有关规定,稳定性好。     

Sarcoplasmic Reticulum Ca2+-ATPase Activity and Glycogen Content in Various Fiber Types after Intensive Exercise in Thoroughbred Horses

To find a new parameter indicating muscle fitness in Thoroughbred horses, we examined time-dependent recovery of glycogen content and sarcoplasmic reticulum (SR) Ca²⁺-ATPase activity of skeletal muscle after intensive treadmill running. Two repeated 50-sec running sessions (13 m/sec) were performed on a flat treadmill (approximately 90%VO₂max). Muscle samples of the middle gluteal muscle were taken before exercise (pre) and 1 min, 20 min, 60 min, and 24 hr after exercise. Muscle fiber type composition was determined in the pre muscle samples by immunohistochemical staining with monoclonal antibody to myosin heavy chain. SR Ca²⁺-ATPase activity of the muscle and glycogen content of each muscle fiber type were determined with biochemical analysis and quantitative histochemical staining, respectively. As compared to the pre value, the glycogen content of each muscle fiber type was reduced by 15–27% at 1 min, 20 min, and 60 min after the exercise and recovered to the pre value at 24 hr after exercise test. These results indicate that 24 hr is enough time to recover glycogen content after short-term intensive exercise. The mean value of the SR Ca²⁺-ATPase activity showed a slight decrease (not significant) immediately after exercise, and complete recovery at 60 min after exercise. There were no significant relationship between the changes in glycogen content of each muscle fiber type and SR Ca²⁺-ATPase. Although further studies are needed, SR Ca²⁺-ATPase is not a useful parameter to detect muscle fitness, at least in Thoroughbred horses.     

Effects of Brief Summer or Winter Fasting on the Muscle Quality of Cultured Pacific Bluefin Tuna (Thunnus orientalis)

The effects of fasting on the quality of the dorsal and ventral ordinary muscles from cultured Pacific bluefin tuna (Thunnus orientalis) during chilled storage were investigated. Tuna were subjected to fasting for 2 days in the summer or 6 days in the winter prior to harvesting. The breaking strength of the dorsal ordinary muscle sampled in the summer increased until 24 h and then decreased. There were no significant differences in the lipid and glycogen content of the ordinary muscle after 9 h of storage between the controls and either fasting group. The pH of the ordinary muscle subjected to summer and winter fasting was higher than in the controls after 24–48 h of storage. However, the relationship between the pH and glycogen content was unclear. The metmyoglobin content during chilled storage was lower in the ordinary muscles from either fasting group than in the controls. In conclusion, fasting for 6 days in the winter improved the color stability of the ordinary muscle without a decline in its lipid content.     

Differences in Muscle Fiber Recruitment Patterns between Continuous and Interval Exercises

We evaluated differences in muscle fiber recruitment patterns between continuous and interval training to develop an optimal training program for Thoroughbred horses. Five well trained female thoroughbred horses (3–4 years old) were used. The horses performed two different exercises on a 10% inclined treadmill: 90%VO₂ max for 4 min (continuous) and 90% VO₂ max for 2 min × 2 times with 10-min interval (interval). Muscle samples were obtained from the middle gluteal muscle before and immediately after the exercises. Four muscle fiber types (type I, IIA, IIA/X, and IIX) were immunohistochemically identified, and the optical density of periodic acid Schiff staining (OD-PAS) in each fiber type and glycogen content of the muscle sample were determined by quantitative histochemical and biochemical procedures, respectively. No significant differences were found in the OD-PASs and glycogen contents between the continuous and interval exercises, but the decreases in OD-PAS of fast-twitch muscle fibers were obvious after interval as compared to continuous exercise. Interval exercise may be a more effective training stimulus for the glycolytic capacity of fast-twitch muscle fiber. The data about muscle fiber recruitment can provide significant insights into the optimal training program not only for thoroughbred horses, but also for human athletes.     

Lysine limitation alters the storage pattern of protein,lipid and glycogen in on‐growing Atlantic salmon

This experiment aimed to test the interaction of lysine limitation with nutrient accretion and muscle carnitine depot in Atlantic salmon. Fish were fed adequate or low‐lysine diets for 3 months. Lysine intake was significantly less (48%) in fish fed the low‐lysine diet as compared with that fed the adequate one. There was no difference in dietary amino acids between treatments, with the exception of lysine. The lower lysine intake was reflected in plasma free lysine being 52% less while the free lysine concentration in the liver and muscle were unaffected. Although there was no significant difference between voluntary feed intakes among treatments, fish fed the low‐lysine diet had reduced growth, protein and energy deposition as compared with fish fed the adequate lysine diet. White trunk muscle contained more glycogen and less protein in fish fed the low‐lysine diet while no difference in lipid was observed. The livers from fish fed the low‐lysine diet contained less glycogen and slightly more fat and protein than the livers from fish fed the adequate lysine diet. Lysine limitation reduced carnitine in the liver without affecting muscle carnitine depot. Thus, low‐lysine diets did not likely affect the fatty acid oxidation capacity. This fact was supported by unaffected fatty acid profiles and lipid classes between treatments during the 3‐month study. In conclusion, lysine limitation does not deplete the muscle carnitine depot during the on‐growing seawater phase of Atlantic salmon, but affects the deposition pattern of nutrients.     

稻瘟菌糖原合成酶激酶MoGSK3功能探究

【目的】观测稻瘟菌（Magnaporthe oryzae）糖原合成酶激酶基因MoGSK3敲除突变体表型,明确MoGSK3在稻瘟菌中的潜在生物学功能,为挖掘防治稻瘟菌新型药剂的潜在靶标提供参考。【方法】基于同源重组原理,用split-PCR方法获得稻瘟菌MoGSK3敲除突变体菌株,将MoGSK3基因克隆到pFL2载体上得到MoGSK3-C融合载体,并将其通过PEG介导的原生质体转化法导入MoGSK3突变体中得到回补菌株。培养观察野生型菌株Guy11、突变体菌株G3-9及回补菌株GC-1的菌落形态和生长状况,采用实时荧光定量PCR(qRT-PCR)检测稻瘟菌产孢相关基因的表达量;观察稻瘟菌不同菌株的分生孢子形态,通过附着胞洋葱表皮穿透试验及接种水稻叶片,研究其致病力;通过KI-I-2染色对野生型菌株Guy11及突变体菌株G3-9分生孢子和附着胞中的糖原运输能力进行观测。【结果】稻瘟菌MoGSK3突变体存在多个表型缺陷,与野生型菌株Guy11相比,MoGSK3突变体菌株G3-9菌落直径显著变小,生长缓慢,产孢相关基因表达量下降且分生孢子出现末端伸长畸形的状态。MoGSK3基因缺失还会导致稻瘟菌菌丝末端无法形成正常的分生孢子梗,附着胞无法穿透洋葱表皮形成侵染菌丝,接种划伤水稻叶片也无法形成褐色病斑。对MoGSK3突变体菌株G3-9分生孢子及附着胞进行糖原染色后发现,突变体菌株G3-9在糖原转运能力方面存在明显缺陷。【结论】MoGSK3基因参与稻瘟菌的生长、分生孢子形成及形态建成、侵染、糖原转运等过程,是稻瘟菌重要的毒力因子。     

铜对肉鸡原代肝细胞活性及糖原含量的影响

为了探讨铜对肉鸡原代肝细胞活性及糖原含量的影响,试验分别用10,50,100μmol/L Cu2+在体外孵育原代肝细胞,PAS染色后测定30分钟和1,2,4,24,48,72小时各时间点的原代肝细胞活性及24小时时的糖原含量。结果表明:50μmol/L和100μmol/L Cu2+仅分别在72小时和4小时之后明显降低细胞活性;随着铜浓度的升高,糖原含量减少。说明高浓度铜可降低原代肝细胞活性,促进肝糖原分解。     

日本血吸虫GSK3蛋白编码cDNA的克隆、表达及其重组蛋白的免疫保护研究

本研究利用生物信息学分析了日本血吸虫(Schistosoma japonicum,sj)Glycogen synthase kinase 3(GSK3)蛋白,并对其中一个GSK3蛋白的编码cDNA进行了克隆和原核表达,制备了特异性的多克隆抗体.同时,还初步评估了重组蛋白的免疫保护效果.生物信息学分析表明在日本血吸虫数据库存在两种GSK3蛋白,且其中一个SjGSK3在日本血吸虫不同发育时期均有转录.Western blot结果表明本研究制备的抗体能特异性识别日本血吸虫SjGSK3重组蛋白,表明该重组蛋白具有良好的免疫原性.动物实验表明免疫SjGSK3重组蛋白的动物与佐剂对照组比较分别获得了平均10.6％减虫率和40.5％肝脏减卵率.