Effect of cortisol and triiodothyronine bath treatments on the digestive enzyme profile and growth of Catla catla larvae during ontogenic development

Digestive enzyme profile is a good indicator of the nutritional and health status of the fish. The present investigation aims to evaluate the effect of exogenous bath treatment of hormones, cortisol and triiodothyronine, on the digestive enzyme activities and growth of carp Catla catla (Ham.) during ontogenic development. Catla larvae (4 days old) were given bath treatment with cortisol (hydrocortisone, 0.2 mg L^?1), 3,5,3′‐triiodothyronine (T3, 2.5 mg L^?1) and a combination of cortisol and T3 for 30 min. Digestive enzyme profile was recorded on every third day and was continued for 30 days. Larvae were fed with live food for initial 14 days and then weaned to mix feeding of live food and prepared diet. Significantly (P < 0.05) higher amylase, total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were found in the hormone‐treated groups compared to the control one during ontogenic development. Among the treated groups, amylase activity was highest in cortisol‐treated larvae. Total protease, trypsin, chymotrypsin, lipase, chitinase and chitinobiase activities were significantly (P < 0.05) higher in larvae exposed at combined treatment of cortisol and T3 compared to the other two groups in most sampling days. Average length, weight and specific growth rate of treated larvae were higher compared to the control one. The combined bath treatment of cortisol and triiodothyronine influenced the digestive enzyme activities of catla larvae and thereby enhanced the growth at early developmental stage. This helps the larvae to overcome the problems associated with early developmental stage.     

Effects of dietary protein to energy ratios on growth,body composition and digestive enzyme activities in Chinese mitten‐handed crab,Eriocheir sinensis

This study aimed to determine the optimal protein to energy ratio (P/E ratio) and evaluate the effect of dietary protein and lipid levels on growth performance, body composition and digestive enzymes activities in Chinese mitten‐handed crab, Eriocheir sinensis. Nine practical diets containing three levels both for protein (DP 30%, 35% and 40%) and lipid (DL 2%, 7% and 12%) with P/E ratios ranging from 13.69 to 19.79 mg KJ^?1 were fed to four replicates of crabs (3.39 ± 0.10 g) for 10 weeks. Weight gain increased significantly with the increase in DP level at each DL level. Moreover, weight gain increased in crabs fed with diets containing DL level from 2% to 12% and DP level from 30% to 35%. However, the diet containing 40% DP and 12% DL levels significantly decreased the growth performance and protein efficiency of the crabs. The whole crab and hepatopancreas lipid contents also increased as dietary lipid increased, but not dietary protein. The total protease activity increased significantly with the increase in dietary protein at each lipid level. The lipase activity was statistically comparable among different DL levels at each DP level. Taken together, the crab fed the diet containing 35% protein and 12% lipid levels with P/E 15.77 mg KJ^?1 revealed optimal growth, feed utilization efficiency and digestive enzymes activities. Moreover, our study indicated that the higher dietary lipid level at a relatively lower dietary protein level could provide protein sparing effect in Eriocheir sinensis.     

温度对波纹龙虾消化酶活力的影响

研究了波纹龙虾温度对不同消化器官中消化酶活力的影响,为人工饲料科学配制依据。用酶学分析方法,设计6个温度梯度(20℃、25℃、30℃、35℃、40℃和45℃),分别测定波纹龙虾胃、肠和肝胰脏的类胃蛋白酶、胰蛋白酶、淀粉酶和脂肪酶的活力。结果显示:在反应温度20℃~45℃范围内,波纹龙虾胃、肠、肝胰脏内的消化酶均随着温度的升高表现为先升后降趋势;不同消化器官中胃蛋白酶活力值出现拐点的温度不一样,胃、肠和肝胰脏的胃蛋白酶活力最大的温度分别是30℃、35℃和40℃;不同消化器官胃蛋白酶的活力有显著差异(P0.05),大小依次为胃肠肝胰脏;波纹龙虾不同消化器官的类胰蛋白酶出现最大酶活力的温度相同,为40℃,但胃的类胰蛋白酶活力明显较肠和肝胰脏的低(P0.05),差值最大可达40 U/mg;波纹龙虾胃、肠和肝胰脏的淀粉酶活力在25℃均出现最大值;在消化器官中,肠道淀粉酶活力最大,与胃和肝胰脏的酶活力有显著性差异(P0.05);波纹龙虾胃、肠和肝胰脏内的脂肪酶活力最大的温度为30℃,活力最高的是肝胰脏,胃内的脂肪酶活力明显的比肠和肝胰脏的要低(P0.05)。     

Exogenous xylanase improves growth,protein digestibility and digestive enzymes activities in Nile tilapia,Oreochromis niloticus,fed different ratios of fish meal to sunflower meal

A 3 × 2 factorial experiment was designed to evaluate the effect of different ratios of fish meal (FM): sunflower meal (SFM) with or without exogenous xylanase supplementation on growth, feed utilization, digestive enzymes activities, apparent digestibility, intestinal and liver morphology and chemical composition of Nile tilapia, Oreochromis niloticus. Three isonitrogenous (329.80 g/kg of crude protein) and isoenergetic (18.46 MJ/kg gross energy) experimental diets were formulated as SFM₁ (FM:SFM = 2:1), SFM₂ (FM:SFM = 1:1) and SFM₃ (FM:SFM = 1:2) based on protein content. Each diet was supplemented with 0 or 0.5 g/kg of exogenous xylanase and was fed to triplicate groups of twelve fish (with initial weight, 1.31 ± 0.02 g) for 84 days. After 84 days of feeding period, the highest weight gain, specific growth rate, protein efficiency, protein productive value and the best feed conversion ratio were recorded in fish fed either SFM₁ or SFM₂ supplemented with exogenous xylanase. Whereas lowest growth performance was recorded in fish fed SFM₂ and SFM₃ un‐supplemented with xylanase. The highest activities of chymotrypsin, trypsin, lipase, amylase, alkaline phosphatase and cholecystokinin were observed in fish fed SFM₁ and SFM₂ diets supplemented with xylanase. The highest ADCs of dry matter, protein, lipid and digestible energy were recorded in fish fed SFM₁ and SFM₂ diets supplemented with exogenous xylanase. Supplementation of exogenous xylanase improved muscularis mucosa thickness, height of mucosal folds and enterocytes of intestinal fish. Addition of exogenous xylanase increased the calcium and phosphorus retention. Results of this study indicated that the addition of exogenous xylanase to diet containing high inclusion level of sunflower meal improved growth, digestive enzymes, nutrient digestibility, histological morphometric of liver and intestine and nutrient retention.     

饲料中添加谷氨酰胺前体物对镜鲤肠道消化酶及Na~+/K~+-ATPase活性的影响

为了研究谷氨酰胺前体物对镜鲤(Cyprinus carpio specularis)肠道消化酶及Na~+/K~+-ATPase活性的影响,分别用谷氨酰胺(Gln)、谷氨酸(Glu)、α-酮戊二酸(AKG)、L-鸟氨酸-α-酮戊二酸(OKG)、L-精氨酸-α-酮戊二酸(AAKG)、α-酮戊二酸钠(2Na-AKG)替代基础饲料中的葡萄糖(添加量为1.5%),配制成6种等氮等能试验饲料,以基础饲料为对照,分别投喂松浦镜鲤(平均体重(40.27±3.96)g),饲养8周后测定镜鲤肠道消化酶及Na~+/K~+-ATPase活性。结果显示:Glu组前肠蛋白酶活性显著高于对照组;Gln组、Glu组、OKG组和AAKG组中肠蛋白酶活性均显著高于对照组。2Na-AKG组前肠脂肪酶活性显著高于对照组和OKG组;2Na-AKG组中肠脂肪酶活性显著高于对照组和Gln组。2Na-AKG组前肠淀粉酶活性均显著高于对照组和Gln组。Gln组和Glu组前肠Na~+/K~+-ATPase活性均显著高于对照组;不同处理组中肠Na~+/K~+-ATPase活性均显著低于对照组;Glu组、AKG组和OKG组后肠Na~+/K~+-ATPase活性均显著高于对照组,Gln组、AAKG组和2NaAKG组后肠Na~+/K~+-ATPase活性则均显著低于对照组。研究表明,饲料中添加Gln、Glu、OKG和AAKG可显著提高鱼体肠道的蛋白酶活性,添加2Na-AKG可显著提高鱼体肠道的淀粉酶和脂肪酶活性。     

生物絮团对罗氏沼虾体组成和消化酶活性的影响

通过向养殖水体中泼洒糖蜜构建生物絮团养殖模式,分析生物絮团营养组成,并探讨生物絮团对罗氏沼虾体组成和消化酶活性的影响。试验分对照组和试验组(生物絮团组),其中试验组在养殖过程中泼洒糖蜜。试验在室内水泥池内(2 m×2 m×0.6 m)进行,每个处理有3个重复,每个重复225尾虾(0.26 g±0.02 g),试验周期为90 d。养殖过程中不换水,糖蜜的泼洒量根据饲料投喂量进行计算(C/N为20)。结果显示:添加糖蜜能够显著促进生物絮团的形成,到第90天时,试验组的絮团体积达21.22 mL/L;而对照组为6.03 mL/L;试验组絮团粗蛋白含量为29.47%,粗脂肪含量为4.32%,二者均显著高于对照组,而粗灰分含量为11.36%,显著低于对照组;泼洒糖蜜对罗氏沼虾体组成的影响不显著,对照组和试验组肌肉粗蛋白含量分别为21.09%和21.20%,粗脂肪含量分别为2.91%和3.06%;另外,向水体中泼洒糖蜜对罗氏沼虾消化酶活性影响显著。试验组罗氏沼虾肠脂肪酶活性、胃脂肪酶活性和胰脂肪酶活性均显著高于对照组;试验组罗氏沼虾糜蛋白酶活性、胰蛋白酶活性也均显著高于对照组。但泼洒糖蜜对肠淀粉酶、胃蛋白酶、胃淀粉酶、胰淀粉酶和纤维素酶活性没有显著影响。试验表明,生物絮团营养组成丰富,能够有效提高消化酶活性。     

Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

饲料异亮氨酸水平对卵形鲳鲹消化酶活性和免疫指标的影响

实验旨在研究饲料异亮氨酸(Ile)水平对卵形鲳鲹(Trachinotus ovatus)消化酶活性和免疫指标的影响。以鱼粉、豆粕、花生粕和L-晶体氨基酸为蛋白源,配制含粗蛋白430 g·kg~(-1)的6种等氮饲料,各组分别含有13.2 g·kg~(-1)、15.7 g·kg~(-1)、18.2 g·kg~(-1)、20.7 g·kg~(-1)、23.2 g·kg~(-1)和25.7g·kg~(-1)(以干物质计)异亮氨酸。每组3个重复,每个重复随机放置20尾鱼[初始体质量(6.36±0.03)g],实验时长8周。结果显示,各实验组的肠淀粉酶和胃蛋白酶活性、血清总抗氧化能力(T-AOC)、溶菌酶(LZM)活性、免疫球蛋白M和补体(C3和C4)浓度均随着饲料Ile水平的升高而先上升后下降,这5个指标的较高值出现在18.2 g·kg~(-1)和20.7 g·kg~(-1)组,而较低值则出现在13.2 g·kg~(-1)和25.7 g·kg~(-1)组,上述4组与其他2组差异显著。血清尿素氮和丙二醛(MDA)浓度的变化趋势则与上述7个指标相反,较低值出现在18.2 g·kg~(-1)和20.7 g·kg~(-1)组,且显著低于其他4组。血清超氧化物歧化酶(SOD)活性则随着饲料Ile水平的增加而显著升高,18.2~25.7 g·kg~(-1)这4组SOD活性没有显著性差异。综上所述,饲料Ile水平为18.2~20.7 g·kg~(-1)时,卵形鲳鲹具有较好的消化性能和免疫机能。     

单宁酸对生长猪胃-小肠仿生消化中消化酶活性及饲粮粗蛋白消化率的影响

本试验旨在探讨单宁酸对生长猪胃、小肠仿生消化中消化酶活性及玉米-豆粕型饲粮干物质和粗蛋白消化率的影响，为评价单宁酸的生物学效应提供参考。试验一采用单因素完全随机设计，考察在无饲粮下2种单宁酸对猪模拟胃液、模拟小肠液消化酶活性的影响。设5个处理，单宁酸添加量分别为0 mg （胃液体积为20 mL，小肠液体积为22 mL）；单宁酸1，10 mg；单宁酸1，20 mg；单宁酸2，10 mg；单宁酸2，20 mg。测定各处理的胃蛋白酶、淀粉酶、胰蛋白酶和糜蛋白酶的活性。试验二考察玉米-豆粕型饲粮添加单宁酸对猪仿生消化中胃、小肠阶段消化酶活性及养分消化率的影响。采用单因素完全随机设计，设5个处理，单宁酸在饲粮中的含量分别为0 mg·（2 g）^-1；单宁酸1，10 mg·（2 g）^-1；单宁酸1，20 mg·（2 g）^-1；单宁酸2，10 mg·（2 g）^-1；单宁酸2，20 mg·（2 g）^-1。测定仿生消化中胃阶段0.5和4 h时胃蛋白酶活性，小肠阶段0.5、4和8 h时淀粉酶、胰蛋白酶和糜蛋白酶活性及生长猪胃-小肠仿生消化测定饲粮的干物质和粗蛋白消化率。结果表明：1）无饲粮的情况下，和空白对照组相比，2种单宁酸对模拟胃液中胃蛋白酶活性无显著影响（P>0.05），单宁酸1比单宁酸2更高地降低了模拟小肠液中淀粉酶、胰蛋白酶和糜蛋白酶的活性（P<0.05）。2）在饲粮进行仿生消化的胃消化0.5～4 h内，除4 h时10 mg·（2 g）^-1添加量外，添加单宁酸1时胃蛋白酶的活性均显著高于添加单宁酸2时的相应值（P<0.05），除单宁酸2在消化0.5 h外，2种单宁酸在添加10 mg·（2 g）^-1时胃蛋白酶活性均显著高于20 mg·（2 g）^-1添加量的相应值（P<0.05）。在小肠仿生消化0.5 h时，饲粮中添加单宁酸1、2的2个水平对消化液中淀粉酶活性无显著影响（P>0.05），但均显著降低了糜蛋白酶的活性（P<0.05）；单宁酸1的消化液中胰蛋白酶活性高于单宁酸2的相应值（P<0.05）。在小肠仿生消化4 h时，除添加水平为20 mg·（2 g）^-1时的糜蛋白酶活性外，饲粮中添加单宁酸1消化液中淀粉酶、糜蛋白酶活性高于添加单宁酸2的相应值，而胰蛋白酶活性低于添加单宁酸2的相应值（P<0.05）。单宁酸1、2的两个添加量均降低了胰蛋白酶和糜蛋白酶的活性（P<0.05）。在小肠仿生消化8 h时，饲粮中单宁酸的添加量影响了淀粉酶的活性，但单宁酸1和单宁酸2各两个添加量在淀粉酶的平均活性上无显著差异（P>0.05）。单宁酸1、2的两个添加量均降低了胰蛋白酶、糜蛋白酶的活性（P<0.05）。3）与对照组相比，两种单宁酸在两种添加水平下均显著降低了饲料粗蛋白消化率（P<0.05），且单宁酸2比单宁酸1更多地降低了饲粮粗蛋白的消化率（P<0.05）。综上所述，在有、无饲粮条件下，单宁酸对消化酶活性呈现不一致影响。单宁酸影响饲粮粗蛋白的消化率可能主要与消化液中糜蛋白酶活性降低以及单宁酸与饲粮中的化学成分形成螯合物降低了小肠消化酶的水解效率有关。     

Nile tilapia broodfish fed high‐protein diets: Digestive enzymes in eggs and larvae

The aim of this study was to assess the activity of gastric, pancreatic and intestinal digestive enzymes during the embryonic and larval development of Nile tilapia (Oreochromis niloticus) GIFT strain Aqua America^® 1 obtained from a broodstock fed two levels of crude protein (CP). A total of 72 females and 24 males, 10 hapas, two CP levels (32% and 38%) and six phases of embryonic (cleavage, blastula, gastrula) and larval (hatching, 7 and 10 days post hatch, dph) stages were used. The eggs were collected in cleavage, blastula and gastrula stages, 300 mg was collected, and kept in cryogenic tubes in liquid nitrogen. For the samples at larval stage, the remaining eggs were separated according to crude protein level and kept in hatcheries and samples were collected on 7 and 10 dph the same way as before. A total of 48 samples were collected: at each protein level (32% and 38% CP), four samples were collected in each phase of embryonic and larval development. Statistical differences were not observed during embryonic development for acid proteases, trypsin, amylase and lipase activity at both levels of crude protein (32 and 38% CP). Differences for acid proteases were noticed on 7 dph; trypsin and amylase on 7 dph and 10 dph. Significant differences on blastula and 7 dph for protease; as for aminopeptidase, there was significant difference on 7 dph. The data indicated early appearance of digestive enzymes in Nile tilapia broodfish receiving 32% CP taking into account the rapid growth and development of this species.