短期饥饿对奥尼罗非鱼幼鱼生长和几种消化酶的影响

研究在(27.0±1)℃条件下,奥尼罗非鱼(Oreochromis niloticus×O.aureus)幼鱼[初始体重为(2.49±0.31)g]短暂饥饿处理后再投喂的补偿生长。结果显示:在恢复生长时期,饥饿3、6、9和12d处理组的特定增长率和摄食率显著高于对照组(P<0.05);各处理组的饲料转化率与对照组差异不显著(P>0.05);胃蛋白酶、胰蛋白酶和脂肪酶的活力变化趋势相同,均随先饥饿再饱食的顺序先下降后上升,而淀粉酶活力变化趋势不明显;恢复投喂后,各组奥尼罗非鱼幼鱼的消化酶活性均恢复到对照组水平。表明饥饿3d组的奥尼罗非鱼幼鱼在恢复生长中具有超补偿能力,饥饿6d组幼鱼具有完全补偿能力,饥饿9和12d组幼鱼具有部分补偿能力。奥尼罗非鱼幼鱼短暂饥饿后出现的补偿生长效应主要通过增加食欲、提高摄食率实现。     

复合酸化剂对鲤生长及消化酶活性的影响

在鲤鱼(Cyprinus carpio)基础饲料中,分别添加不同梯度的复合酸化剂(质量分数分别为0、0.2%、0.3%),研究复合酸化剂对其生长和消化酶活性的影响。结果表明:1)添加复合酸化剂显著(P<0.05)提高鲤鱼的相对增重率、特定生长率、饵料转化效率、饲料蛋白质效率以及饲料蛋白质沉积率等生长性能指标;2)添加质量分数0.3%的复合酸化剂时,鲤鱼肝胰脏、前肠和中肠的蛋白酶活性显著提高(P<0.05),酸化剂组的淀粉酶和脂肪酶活性有升高的趋势,但与对照组差异不显著;3)鱼体的粗蛋白质、粗脂肪、粗灰分、水分、肥满度及内脏比等品质指标在各组间差异不显著。综上所述,鲤鱼饲料中添加以柠檬酸、乳酸、磷酸等为主的复合酸化剂,可显著提高鲤鱼肝胰脏、前肠和中肠的蛋白酶活性,促进鲤鱼生长。     

罗非鱼开口饵料的初步研究

研究了配合饵料、轮虫、蛋黄、轮虫+人工配合饵料(3～12日龄投喂轮虫,13日龄起投喂配合饵料)等4种开口饵料对奥尼罗非鱼仔鱼生长、发育和存活影响。结果表明,1)投喂不同饵料对奥尼罗非鱼仔鱼生长具有显著影响(P<0.05),轮虫+配合饵料组生长较快,绝对增重率和特定增重率分别为0.011g/d和17.18%d-1;蛋黄组生长最慢,绝对增重率和特定增重率与其他各组差异显著(P<0.05)。2)轮虫+配合饵料与轮虫组的仔鱼成活率较高,分别为90.3%和89.7%,2组差异不显著(P>0.05),且仔鱼发育较好,大部分个体进入稚鱼阶段;蛋黄组成活率最低,只有40.7%,个体发育较为缓慢,与其他各组差异显著(P<0.05)。可见,投喂鲜活的天然饵料(轮虫)+配合饵料是罗非鱼苗种培育的最佳开口饵料。     

草鱼、鲤鱼肠道、肝胰脏消化酶活性的初步研究

用两种不同蛋白质和糖类水平的饲料喂养草鱼、鲤鱼夏花及一龄鱼种，通过对两种鱼肝胰脏及肠道的消化酶活性测定发现，在一定温度条件下，随着水温升高，两种鱼肠道及肝胰脏的蛋白酶和淀粉酶比活均升高；鱼类摄食后六小时酶比活明显大于摄食后二十四小时的酶比活，在水温２５℃条件下，用两种不同饲料分别饲养草鱼、鲤鱼六周后发现：随着饲料蛋白质水平的提高，草鱼、鲤鱼肝胰脏中蛋白酶比活明显升高鲤，而总肠变化不显著；随饲料中糖量水平的提高，草鱼肝胰脏及肠道的淀粉酶比活明显升高，而鲤鱼差异不显著．草鱼，鲤鱼肠道蛋白酶比活以后肠最高，中肠次之，前肠最小。而肠道淀粉酶比活性却随饲料成分而发生差异。两种鱼肝胰脏消化酶活性均为一龄鱼种大于夏花阶段，这与组织切片的观察结果相符。     

三氯生对雌性斑马鱼肝胰脏凋亡相关基因表达的影响

【目的】研究三氯生(TCS)对雌性硬骨鱼类肝胰脏损伤的相关分子机制。【方法】采用半静态水体接触染毒法,将雌性斑马鱼(Danio rerio)暴露于0、0.017、0.034、0.068 mg/L的TCS溶液42 d,采用普通PCR和实时荧光定量PCR技术测定雌性斑马鱼肝胰脏凋亡相关基因(Bcl-2、p53、MDM2、Bax)的表达情况。【结果】0.017~0.068 mg/L组的雌性斑马鱼肝胰脏Bcl-2、MDM2和Bax基因的表达极显著下调(P<0.01),p53基因的表达极显著上调(P<0.01)。【结论】0.017~0.068 mg/L的TCS显著影响雌性斑马鱼肝胰脏凋亡相关基因的表达,促进雌性斑马鱼肝胰脏细胞凋亡的发生。     

尼罗罗非鱼干扰素调节因子4基因的克隆与表达

【目的】探究干扰素调节因子(IRF)在尼罗罗非鱼(Oreochromisniloticus)免疫中的作用。【方法】从尼罗罗非鱼中鉴定IRF4同源物,命名为OnIRF4。使用生物信息学分析软件分析OnIRF4的氨基酸序列。对健康尼罗罗非鱼腹腔注射浓度为3×10~7 CFU/mL的无乳链球菌200μL,用实时荧光定量PCR法分析OnIRF4在健康鱼中的表达模式及其对细菌感染的生物学效应。【结果与结论】OnIRF4序列的开放阅读框为1 350 bp,编码449个氨基酸。OnIRF4与其他物种的IRF4蛋白的同源性为52%～98%。推导的OnIRF4肽链具有干扰素因子典型的DNA结合结构域(DBD)、干扰素相联结构域(IAD)。亚细胞定位结果表明,OnIRF4主要定位在细胞核。表达分析表明,OnIRF4在健康尼罗罗非鱼皮肤、头肾、鳃、脑、肠、肌肉、肝、胸腺和脾中均有分布。在无乳链球菌(Streptococcus agalactiae)感染后,OnIRF4在肝脏、脾脏、头肾、脑中的表达显著上调(P0.05),表明OnIRF4参与了尼罗罗非鱼对细菌感染的免疫反应。     

盐度对尼罗罗非鱼血清甲状腺素浓度和成活率的影响

在尼罗罗非鱼(Oreochromis niloticus)由淡水逐渐向不同盐度海水(5,10,15,20,25)适应过程中,运用放射免疫测定法研究血清中三碘甲腺原氨酸(T3)浓度的变化,并分析盐度变化对尼罗罗非鱼成活率的影响。结果表明,盐度对尼罗罗非鱼血清中T3水平和成活率有着显著的影响。在盐度5~20的范围内变化后,血清T3的浓度在6 d内随着盐度上升而增大,第7天达到高峰;随后,逐渐降低并趋于稳定,在第14天时,各盐度组尼罗罗非鱼血清中T3开始处于稳定状态;稳定后,各实验组血清中T3的浓度均比对照组升高,并表现出随盐度的升高而升高的趋势。在盐度20时,实验第17天,尼罗罗非鱼的成活率为80%。而在盐度为25时,实验第11天,尼罗罗非鱼的成活率仅为54%。对不同盐度下尼罗罗非鱼死亡原因及血清甲状腺激素在盐度变化下的生理意义和对死亡率的影响作了讨论。     

温度对尼罗罗非鱼急性耐盐性的影响

采用两因素交叉分组的方法,初步研究了温度对尼罗罗非鱼急性耐盐性的影响。结果表明:温度与盐度的交互作用非常明显(P<0.05),在温度为25、28、31℃时,尼罗罗非鱼的96h半致死盐度(MSL-96)分别为26.2、22.5、16.6,三者之间差异显著(P<0.05),表现出随温度升高而MSL-96降低的趋势。不同盐度和温度水平的平均成活时间(MST)均差异显著(P<0.05),同时也表现出随温度和盐度升高而MST降低的趋势;在同一温度下各盐度水平的50%死亡时间(ST50)差异显著(P<0.05)。     

不同温度下罗非鱼人工感染海豚链球菌后血液生化指标及组织病理

在28℃、33℃两种温度下,用海豚链球菌人工感染尼罗罗非鱼后,测定尼罗罗非鱼血液生化指标。结果显示:两个温度水平病鱼的血清总蛋白(TP)随时间延长都呈现升高趋势;谷丙转氨酶(ALT)、谷草转氨酶(AST)活性随时间延长先升高后降低再升高,33℃实验组ALT、AST活性比28℃实验组达峰值时间短,并显著高于对照组(p<0.05);尿素、钾(K+)含量随时间延长逐渐升高后降低,钠(Na+)含量先升高后降低再升高,肌酐含量呈上升趋势;血清碱性磷酸酶(AKP)活性28℃实验组呈先升高后降低趋势,33℃实验组呈升高趋势;超氧化歧化酶(SOD)活性随时间延长先升高后降低再升高,33℃实验组各个时间点基本高于28℃实验组;球蛋白(GLO)随感染后时间延长呈现升高趋势,至120 h,显著高于对照组(p<0.05),但温度组间各时间点没有显著差异(p>0.05)。组织病理学观察发现,33℃实验组肝细胞、肾小管上皮细胞水肿变性;28℃实验组肝细胞出现嗜酸性小体;肾小管上皮细胞出现嗜酸性的红色小体,颗粒变性。表明感染海豚链球菌的尼罗罗非鱼肝脏、肾脏均有损伤,造成功能障碍,高温时损伤更严重。     

恩诺沙星在凡纳滨对虾体内的代谢和残留消除规律

在26±2℃水温下,每天投喂含有恩诺沙星药物的饲料,研究恩诺沙星在凡纳滨对虾Litopenaeus vanname肌肉、肠和肝胰脏组织中的代谢和残留消除规律。残留药物用乙腈提取,液相色谱串联质谱仪检测。结果表明:恩诺沙星在凡纳滨对虾体内可代谢为环丙沙星,对虾体内同时有恩诺沙星和环丙沙星两种药物残留;环丙沙星在肌肉、肝胰脏和肠组织中的消除时间分别为6、8、10 d,而恩诺沙星在这三组织中的消除时间则为12、14、16 d。建议把肠作为该药残留监控的靶组织,凡纳滨对虾的休药期不少于16 d。     

Effects of temperature, pH and NaC1 on protease activity in digestive tract of young turbot, Scophthalmus maximus

The protease activity in digestive tract of young turbot Scophthalmus maximum was studied, and the optimal pH, temperature and NaCl concentration were determined for different portions of the fish＇s internal organs. The optimal activity in the fish＇s stomach was at pH of 2.2, while that in the intestinal extracts was within the alkaline range from 9.5 to 10.0. In hepatopancreas, the optimal pH was in low alkalinity at 8.5. The optimal reaction temperature was above 40℃ in stomach, intestine and hepatopancreas. With increasing temperature, the pH value increased in stomach, while in the intestine, an opposite tendency was observed due to combined effect of pH and temperature. NaCl concentration showed inhibitory impact on protein digestion in hepatopancreas. The main protease for protein digestion in turbot seemed to be pepsin. Moreover, the maximum protease activity in different segments of intestine existed in the hindgut.     

Digestive enzyme activities in mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis

The mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis occupy the intertidal zone. However, both species have their own unique diet. The former is an herbivore and the latter is a carnivore. In order to reveal the relationship between digestive enzyme activities and diets in the two species, the activities of protease (P), non-specific bile salt-activated lipase (BAL) and α-amylase (A) were determined in the stomach and intestine of adult mudskipper B. pectinirostris and Chinese black sleeper B. sinensis. The results showed that the activities of protease, BAL and α-amylase in the intestine of B. pectinirostris were significantly (P<0.05) higher than those in the stomach. In B. sinensis, gastric protease activity was not different from the intestinal protease (P>0.05), while BAL and α-amylase activities of the intestine were significantly (P<0.05) higher than those of the stomach. The activity of gastric protease in B. sinensis was significantly (P<0.05) higher than that in B. pectinirostris, while the activities of intestinal protease were not different between the two fish species (P>0.05). BAL activities of the stomach and intestine in B. sinensis were significantly (P<0.05) higher than those in B. pectinirostris, while α-amylase activities of the stomach and intestine in B. pectinirostris were significantly (P<0.05) higher than those in B. sinensis. The ratios of P/BAL, A/P and A/BAL of the digestive tract in B. pectinirostris were 1.5, 107.3 and 158.6, respectively; and those in B. sinensis were 0.2, 1.6 and 0.2, respectively. It can be concluded that food digestion in the adult B. pectinirostris is mainly carried out in the intestine, whereas in the adult B. sinensis it is initiated in the stomach and finishes in the intestine. The activities of BAL and α-amylase in B. pectinirostris and B. sinensis are well correlated with their diets. However, a clear-cut correlation between protease activity and diets is not found in these two species.     

高淀粉水平下精氨酸对松浦镜鲤肠道形态与功能的影响

【目的】探讨高淀粉饲料中添加精氨酸对松浦镜鲤(CyprinuscarpioL.)肠道消化酶活性及肠道形态的影响。【方法】选用初始体质量(6.84±0.02)g的松浦镜鲤360尾,随机分为5组,每组4个重复,每个重复18尾鱼。实验饲料分为5组,分别在45%水平的高淀粉饲料中添加质量分数0%、0.6%、1.2%、1.8%、2.4%的精氨酸,实验周期为8周,进行肠道消化酶和形态的测定。【结果】前、中、后肠的淀粉酶活性,各组之间没有显著差异(P> 0.05);添加2.4%精氨酸组的前肠脂肪酶活性显著高于对照组(P <0.05),中肠和后肠的脂肪酶活性各组间没有显著差异(P>0.05);前、中、后肠蛋白酶活性随精氨酸添加量的增加而增加,添加1.8%和2.4%精氨酸组的前肠和中肠蛋白酶活性显著高于对照组(P<0.05),添加2.4%精氨酸组后肠的蛋白酶活性显著高于对照组(P<0.05)。高淀粉饲料中添加Arg显著影响了肠道皱襞高度,但对肠绒毛宽度和肌层厚度没有显著影响。2.4%精氨酸组前肠皱襞高度显著高于其他各组(P <0.05),1.8%精氨酸组中肠和后肠皱襞高度显著高于对照组(P <0.05)。1.2%、1.8%和2.4%组肠体指数显著高于对照组(P <0.05),2.4%精氨酸组肠长指数显著高于对照组(P <0.05)。各组前、中、后肠肠绒毛密度没有显著差异,且排列较整齐。【结论】高淀粉饲料中添加1.8%~2.4%精氨酸可以增加松浦镜鲤肠道消化酶活性,同时改善了松浦镜鲤的肠道形态结构。     

饲喂频率对奥尼罗非鱼仔稚鱼生长、体成分和消化酶活力的影响

研究在(28.0±1)℃条件下,5种不同饲喂频率对奥尼罗非鱼(Oreochromis niloticus×O.aureus)仔稚鱼的生长、体成分及几种消化酶活力的影响。结果显示:随着饲喂频率的增加,奥尼罗非鱼仔稚鱼的摄食率和特定生长率逐步提高,1次/d组的摄食率和特定生长率显著低于其他组(P<0.05);2、3、4、5次/d组的相对增重率之间差异不显著(P>0.05),但显著高于1次/d组;各组的饲料转化率随饲喂频率的增加而降低;胃蛋白酶、脂肪酶和淀粉酶活力随饲喂频率的增加而显著降低(P<0.05);各组胰蛋白酶变化差异不明显(P>0.05);随饲喂频率的增加,奥尼罗非鱼仔稚鱼的灰分和蛋白质含量无显著变化(P>0.05);脂肪含量显著上升,水分含量则明显下降(P<0.05)。分析表明,奥尼罗非鱼仔稚鱼最适饲喂频率为3次/d。     

饥饿对褐菖鮋代谢水平的影响

在水温18℃±1℃和盐度27.0±0.5条件下,研究了饥饿对褐菖鮋耗氧率和排氨率的影响。结果表明:对照组的平均耗氧率为1.082 mg.g-1.h-1;饥饿组耗氧率随着饥饿时间的延长而下降,饥饿期间,褐菖鲉的平均耗氧率为0.893 mg.g-1.h-1,总体上低于对照组。排氨率随着饥饿时间的延长下降趋势呈阶段性变化:饥饿1 d、3 d后的排氨率略低于对照组但差异不显著(P>0.05);饥饿3～7 d,排氨率下降幅度变大,饥饿7d较饥饿3 d的排氨率下降了185.075%;饥饿14 d、21 d后排氨率分别为0.053 mg.g-1.h-1和0.028 mg.g-1.h-1,呈继续下降趋势,但下降幅度减缓。褐菖鮋的nO∶nN比值始小幅度上升,7 d时开始迅速上升,21 d后升高至27.754,差异显著(P<0.05)。     

Effect of dietary genistein on growth performance,digestive enzyme activity,and body composition of Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

An 8-week feeding experiment was performed to evaluate the effect of dietary genistein on growth performance, body composition, and digestive enzymes activity of juvenile Nile tilapia (Oreochromis niloticus). Four isonitrogenous and isoenergetic diets were formulated containing four graded supplements of genistein: 0, 30, 300, and 3 000 μg/g. Each diet was randomly assigned in triplicate to tanks stocked with 15 juvenile tilapia (10.47±1.24 g). The results show that 30 and 300 μg/g dietary genistein had no significant effect on growth performance of Nile tilapia, but the higher level of genistein (3 000 μg/g) significantly depressed the final body weight and specific growth rate. There was no significant difference in survival rate, feed intake, feed efficiency ratio or whole body composition among all dietary treatments. An assay of digestive enzymes showed that the diet containing 3 000 μg/ggenistein decreased stomach and hepatopancreas protease activity, and amylase activity in the liver and intestine, while a dietary level of 300 μg/g genistein depressed stomach protease and intestine amylase activities. However, no significant difference in stomach amylase activity was found among dietary treatments. Overall, the results of the present study indicate that a high level of dietary genistein (3 000 μg/g, or above) would significantly reduce the growth of Nile tilapia, partly because of its inhibitory effect on the activity of major digestive enzymes. Accordingly, the detrimental effects of genistein, as found in soybean products, should not be ignored when applied as an alternative ingredient source in aquaculture.     

Effects of Dietary Stachyose on Growth Performance,Digestive Enzyme Activities and Intestinal Morphology of Juvenile Turbot(Scophthalmus maximus L)

A 12-week feeding trial was conducted to evaluate the effects of dietary stachyose on the growth performance, digestive enzymes activities and intestinal structures of juvenile turbot(Scophthalmus maximus L). Five isonitrogenous(49.58% crude protein) and isolipidic(10.50% crude lipid) diets were formulated to contain 0(Control), 0.625%(S-0.625), 1.25%(S-1.25), 2.5%(S-2.5) and 5%(S-5) stachyose, respectively. With the increase of stachyose level, the growth performance and feed utilization of turbot, such as the specific growth rate, final mean body weight, weight gain rate and feed efficiency, increased significantly(P 0.05) and then stabilized. The feed intake of fish fed S-5 was significantly higher(P 0.05) than that of fish in other groups. The activities of trypsin, intestinal caseinolytic, stomach and intestinal amylase were significantly influenced by stachyose(P 0.05). The highest values of trypsin and intestinal caseinolytic activities were observed in group S-1.25, while the highest activity of stomach amylase and the lowest activity of intestine amylase were observed in group S-5. No lesion or damage was found on the distal intestine structures of fish from all treatments, while the height of simple folds in the distal intestine was significantly increased(P 0.05) when 1.25% or 2.5% stachyose was added in the diets. These results indicated that moderate level of stachyose(1.25%) improves the growth performance, feed utilization, digestive enzyme activities and the distal intestine structures of juvenile turbot.     

Effects of starving and re-feeding strategies on the growth performance and physiological characteristics of the juvenile tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>)

Starving and re-feeding trials were conducted to evaluate the effects of starvation duration and recovery times on the growth performance and physiological characteristics of juvenile tongue sole Cynoglossus semilaevis, which included growth, body composition, intestinal morphology and digestive enzymes activities. The fish starved for one (D1), two (D2) and four (D4) days, respectively, were recovered for three (R3, R6, R12), seven (R7, R14, R28) and eleven (R11, R22, R44) folds of starvation days while the control fish were fed constantly. The experiment was conducted for 96 days. The fish individuals in D1R11, D2R14 and D2R22 caught up in weight with the control, indicating that complete compensatory growth existed in these fish individuals. The highest body weight gain and total specific growth rates were observed in D2R22. Food intake increased significantly in most treatments in comparison with that of the control except for D1R7 and D2R22 (P < 0.05). Food conversion efficiency and apparent digest coefficient in D2R22 were significantly promoted, being higher than that of the control (P < 0.05). The activities of trypsin and lipase were found to be closely related to the growth performance. The activities of liver and hindgut trypsin, also midgut and hindgut lipase in D2R22 were significantly higher than those of the control (P < 0.05). The fold height (HF) of foregut and midgut increased significantly in D2R22, and HF decreased significantly in D1R3, D2R6 and D4R12 (P < 0.05). On our findings, we may conclude that the optimum starving and re-feeding strategy is starving for 2 days and re-feeding for 22 days. And in this starving and re-feeding strategy, the compensatory growth could be mostly attributed to the promotion of food conversion efficiency and digestibility coefficiency.     

Effects of Starving and Re-feeding Strategies on the Growth Performance and Physiological Characteristics of the Juvenile Tongue Sole(Cynoglossus semilaevis)

Starving and re-feeding trials were conducted to evaluate the effects of starvation duration and recovery times on the growth performance and physiological characteristics of juvenile tongue sole Cynoglossus semilaevis, which included growth, body composition, intestinal morphology and digestive enzymes activities. The fish starved for one(D1), two(D2) and four(D4) days, respectively, were recovered for three(R3, R6, R12), seven(R7, R14, R28) and eleven(R11, R22, R44) folds of starvation days while the control fish were fed constantly. The experiment was conducted for 96 days. The fish individuals in D1R11, D2R14 and D2R22 caught up in weight with the control, indicating that complete compensatory growth existed in these fish individuals. The highest body weight gain and total specific growth rates were observed in D2R22. Food intake increased significantly in most treatments in comparison with that of the control except for D1R7 and D2R22(P 0.05). Food conversion efficiency and apparent digest coefficient in D2R22 were significantly promoted, being higher than that of the control(P 0.05). The activities of trypsin and lipase were found to be closely related to the growth performance. The activities of liver and hindgut trypsin, also midgut and hindgut lipase in D2R22 were significantly higher than those of the control(P 0.05). The fold height(HF) of foregut and midgut increased significantly in D2R22, and HF decreased significantly in D1R3, D2R6 and D4R12(P 0.05). On our findings, we may conclude that the optimum starving and re-feeding strategy is starving for 2 days and re-feeding for 22 days. And in this starving and re-feeding strategy, the compensatory growth could be mostly attributed to the promotion of food conversion efficiency and digestibility coefficiency.