长薄鳅幼鱼消化道指数和消化酶活性研究

测定了长薄鳅(Leptobotia elongate Bleeker)幼鱼(8.0～110.1 g)的消化道指数和不同体重组(10 g、30 g、50 g)幼鱼的胃、肝胰脏、前肠、中肠、后肠的淀粉酶、胃蛋白酶、胰蛋白酶及脂肪酶活性。结果:(1)长薄鳅幼鱼肝指数平均为(1.400±0.004)%,比肠长平均为(0.418±0.080),体重与体长呈幂函数相关,相关式为:W=0.019L2.818 1(R2=0.948,P<0.01,n=43);鱼体肥满度在30 g组中最高。(2)各部位消化酶活性均以蛋白酶>脂肪酶>淀粉酶;各体重组淀粉酶活性在肝胰脏中最高(P<0.05);蛋白酶活性在胃和中肠中较高;脂肪酶活性沿消化道从前至后升高;(3)各部位淀粉酶活性在各体重组间变化不大,胃蛋白酶、肠段胰蛋白酶和脂肪酶活性随体重增加而下降。本研究认为长薄鳅幼鱼为杂食、偏肉食性,其消化酶活性存在组织特异性,体重30 g左右是其营养需求转变或者生长发育的重要时期。     

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)。     

温度和pH值对河鲈消化酶活性的影响

研究了离体状态下河鲈(Perca fluviatilis)体内淀粉酶、蛋白酶和脂肪酶在不同温度和pH时的活性变化。结果显示:在河鲈肝胰脏、肠道、胃3部位,淀粉酶的最适温度均为30℃,最适pH值分别为5.0、7.0、8.0;脂肪酶的最适温度均为40℃,最适pH值分别为4.0、3.0、6.0;肝胰脏、肠道蛋白酶的最适温度为50℃,胃蛋白酶的最适温度为40℃。肝胰脏、肠和胃蛋白酶的最适pH值分别6.0、8.0、2.0。在各自最适温度下,肠道中脂肪酶比活力最高,其次是肝胰脏和胃;肝胰脏淀粉酶比活力最高,其次是肠道和胃;肠道中的蛋白酶比活力最高,其次是胃和肝胰脏。     

不同水平的蚯蚓和蚯蚓粪对草鱼消化能力的影响

本研究首次在基础饲料中添加不同浓度的蚯蚓(5%、7.5%、10%)和蚯蚓粪(2%、4%、6%),饲养平均体重(54.03±0.15)g的草鱼100d,研究蚯蚓和蚯蚓粪对草鱼消化能力的影响。结果表明:添加7.5%～10%蚯蚓和6%蚯蚓粪显著提高了草鱼肝胰脏和前肠中胃蛋白酶、胰蛋白酶、淀粉酶和脂肪酶的活性,以添加7.5%蚯蚓效果最好;采用石蜡切片技术观察各试验组草鱼前肠肠道内部结构,结果表明,添加7.5%～10%蚯蚓可显著提高草鱼肠道褶皱高度、宽度和基层厚度。综上所述,在提高草鱼消化能力方面,以饲料中添加7.5%蚯蚓的效果最好。     

青海湖裸鲤和花斑裸鲤消化酶活力对比

本实验对比研究青海湖裸鲤(Gymnocypris przewalskii)和花斑裸鲤(Gymnocypris eckloni)消化酶活性差异。两种鱼各挑选9尾体重100g左右的鱼作为实验对象,分别检测肝胰脏、前肠、中肠及后肠中的脂肪酶、淀粉酶、糜蛋白酶和胰蛋白酶活性。结果表明:作为无胃鱼,两种鱼肠道消化酶活性显著高于肝胰脏,其中前肠是主要的消化器官,后肠具有较高的蛋白质消化能力。通过对比研究发现青海湖裸鲤具有较高的脂肪和糖类消化能力,而花斑裸鲤具有较高的蛋白质消化能力。     

幼鲤肝胰脏、肠道消化酶活性比较研究

采用同日龄幼鲤在同一饲养条件下，连续测得38、76、114日龄时肝胰脏、肠道蛋白酶、脂肪酸和淀粉酶活性。结果表明：肝胰脏、肠道蛋白酶活性，肝胰脏淀粉酶活性随日龄增加而增加；38至76日龄时肠道脂肪酶、淀粉酶活性明显增加，76至114日龄变化不明显，而肝胰脏脂肪酶活性显著增加；38至76日龄时肠道蛋白酶、脂肪酶、淀粉酶活性高于肝胰脏，114日龄时肝胰脏蛋白酶、脂肪酶活性高于肠道，淀粉酶活性与肠道接近。     

澳洲宝石鱼营养成分和消化酶活性的研究

采用多重比较法分析澳洲宝石鱼肝胰脏、肠道、胃组织的淀粉酶、蛋白酶和脂肪酶活性以及肌肉的营养成份。结果表明,宝石鱼肌肉中蛋白质含量为17.87%,脂肪含量为8.05%,糖类含量为3.21%,灰分含量为2.83%,水分含量为70.29%。说明宝石鱼是营养价值较高的鱼种。三种消化酶活力分布不同,淀粉酶、蛋白酶比活力大小均为:肠道>胃>肝胰脏,且三者间均存在极显著差异(P<0.01),而胃蛋白酶活力(pH3.0)最高;脂肪酶比活力在胃、肠道不存在显著性差异,但极明显高于肝胰脏(P<0.01)。三种消化酶在同一消化器官内的活性,肝胰脏、肠道和胃均表现为蛋白酶>淀粉酶>脂肪酶,A/P比值均小于1。表明澳洲宝石鱼为肉食性或偏肉食性鱼类。     

翘嘴红鲌胃、肠道及肝胰脏主要消化酶活力的研究

对翘嘴红鲌胃肠道及肝胰脏的淀粉酶、脂肪酶、蛋白酶3种主要消化酶活力进行初步研究。结果表明:胃、肠道、肝胰脏的淀粉酶的最适pH均为6.0;脂肪酶的最适pH分别为8.0、8.0、7.5;胃蛋白酶最适pH为2.0,肠道、肝胰脏蛋白酶的最适pH分别为7.0、7.5。同时研究了在最适pH条件下,不同反应温度对3种主要消化酶的活性影响。结果表明:翘嘴红鲌淀粉酶在肝胰脏、肠道、胃的最适温度分别为30℃,30℃,25℃;脂肪酶的最适温度分别为40℃,30℃,35℃;蛋白酶的最适温度分别为50℃,50℃,40℃。在一定的温度范围内,3种消化酶的活力均呈先上升后下降趋势。最适反应温度下翘嘴红鲌脂肪酶与淀粉酶的活力分布均呈现肝胰脏>肠道>胃,蛋白酶活力肠道与胃接近,大于肝胰脏蛋白酶活力。     

饲料蛋白水平对匙吻鲟幼鱼肌肉营养成分及消化酶活力的影响

配制蛋白水平34.16%、36.53%、39.10%和41.08%的4种等能半精制饲料,在水温28～32℃条件下,饲喂体质量(19.91±3.73)g的匙吻鲟45d,研究饲料蛋白水平对匙吻鲟肌肉营养成分和消化酶(酸性蛋白酶、碱性蛋白酶、α-淀粉酶和脂肪酶)活性的影响。结果显示,41.08%组体质量显著高于36.53%组,其余各组体质量差异不显著;肌肉粗蛋白含量随着饲料蛋白水平的增加而增加,各组肌肉水分、粗脂肪和粗灰分含量差异不显著;酸性蛋白酶和肝胰脏碱性蛋白酶活力随饲料蛋白水平的增加而增加,而肠道碱性蛋白酶活力随饲料蛋白水平的增加而降低;消化道α-淀粉酶活力随饲料蛋白水平的增加而降低,而肝胰脏α-淀粉酶活力表现出相反的趋势;饲料蛋白水平对消化道和肝胰脏脂肪酶活力影响不显著,各组脂肪酶活力差异不显著。试验结果表明,饲料蛋白水平对匙吻鲟肌肉粗蛋白含量、酸性蛋白酶、碱性蛋白酶和α-淀粉酶都会产生显著影响;饲料蛋白水平为39.10%比较适宜。     

不同生长阶段宝石鲈消化酶活性的初步研究

测定了出膜后30 d、70 d、120 d至成鱼4种规格,体重分别为(2.29±0.49)g、(23.16±4.23)g、(88.06±6.73)g、(270.26±9.17)g的宝石鲈(Scorturm barcoo)胃、肝胰脏、前肠、中肠、后肠的淀粉酶、胃蛋白酶和脂肪酶的活性。结果表明,随着宝石鲈的生长,淀粉酶在各生长阶段活力均很小,肝胰脏、前肠和后肠中的淀粉酶活力表现出逐渐增大的趋势;胃中的胃蛋白酶活性表现为逐渐增高,至规格3后胃蛋白酶活性趋于稳定;肠道是脂肪消化的主要场所,其中前肠脂肪酶变化趋势表现为先上升后下降,在前肠脂肪酶下降后中肠脂肪酶升高,起到了继续消化脂肪的作用。     

不同蛋白能量比饲料对草鱼幼鱼消化酶活性的影响

以鱼粉和豆粕为蛋白源,鱼油和豆油等比例混合油为脂肪源,共配制4个蛋白水平(20%、25%、30%、35%),每一蛋白水平设4个脂肪水平(4.5%、8%、11.5%、15%),共16组饲料。每组设3个重复,在水温23~29℃下,连续投喂初始体重为(16.84±0.28)g的草鱼(Ctenopharyngodon idellus)10周,研究不同蛋白能量比饲料对草鱼幼鱼消化酶活性的影响。结果显示:肠道蛋白酶的活性随着饲料蛋白含量的升高而显著升高(P<0.05),饲料蛋白水平在20%~30%时,饲料中的脂肪水平对蛋白酶活性影响不显著(P>0.05),蛋白水平在35%时,高脂肪抑制了蛋白酶活性,肝胰脏的蛋白酶变化规律类似于肠道;肠道和肝胰脏淀粉酶活性在饲料蛋白含量相同的情况下,随着脂肪含量的升高呈现显著的下降趋势(P<0.05),但饲料中蛋白(30%,35%)和脂肪(8%,11.5%,15%)含量较高时,淀粉酶活性较稳定;肠道和肝胰脏的脂肪酶活性在饲料蛋白含量相同的情况下,随着脂肪含量的升高,呈显著上升的趋势(P<0.05),但在蛋白含量(30%,35%)和脂肪含量(8%、11.5%,15%)的6个试验组之间差异不显著(P>0.05);在脂肪含量分别为4.5%、8%和11.5%时,饲料的蛋白含量对肠道的脂肪酶活性无显著性影响(P>0.05),当脂肪含量为15%时,随着蛋白含量的升高,肠道的脂肪酶活性显著降低(P<0.05)。结果表明,草鱼饲料中蛋白含量为30%和脂肪为8%(即蛋白能量比21.7 mg/kJ)的试验组草鱼获得较高的消化酶活性。     

Effects of lysine and methionine supplementation on growth,body composition and digestive function of grass carp (Ctenopharyngodon idella) fed plant protein diets using high‐level canola meal

A 76‐day feeding trial was carried out to evaluate the effects of Lysine and Methionine supplementation on growth and digestive capacity of grass carp (Ctenopharyngodon idella) fed plant protein diets using high‐level canola meal (CM). Fish with initial average weight 103.9 ± 0.6 g were fed three extruded diets. Fish meal (FM) diet was formulated as the normal control with 40 g kg^?1 FM and 300 g kg^?1 CM; CM diet was prepared by replacing all FM with CM (total 340 g kg^?1) without Lys or Met supplementation; CM supplement (CMS) diet was similar to CM diet but was supplemented with essential amino acids (EAA) to ensure the levels of Lys and Met similar to those in the FM diet. Feed intake, feed efficiency and specific growth rate of the grass carp fed CMS and FM diets were similar (P > 0.05), but higher than those of the grass carp fed CM diet (P < 0.05). The hepatosomatic index, relative gut length, intestosomatic index and intestinal folds height were significantly improved in fish fed FM and CMS diets as compared to CM diet (P < 0.05). Lower activities of trypsin, lipase and amylase in hepatopancreas were observed in fish fed CM diet (P < 0.05). Three hundred and forty gram per kilogram CM without Lys or Met supplementation significantly decreased trypsin, lipase and amylase mRNA levels in hepatopancreas (P < 0.05). These results indicated that the high supply of CM (340 g kg^?1) in plant protein (200 g kg^?1 soybean meal and 100 g kg^?1 cottonseed meal) diets decreased digestive ability through decreasing digestive enzyme activities and enzyme gene's expressions of grass carp, and these side effects can be reversed by supplementing Lys and Met. Therefore, CM could be high level used in a plant protein blend‐based extruded diet for grass carp as long as EAA were supplemented.     

鲢、鳙蛋白酶、淀粉酶的研究

运用酶动力学方法研究了鲢、鳙的蛋白酶和淀粉酶的活性分布。鲢、鳙的蛋白酶活性在肝胰脏中最强,脾脏中也较强,而在胆汁中未测出。该酶活性在肠粘膜的第Ⅱ段最大,第Ⅰ段向第Ⅱ段递增,第Ⅱ段向肠后端递减,但第Ⅴ段的该酶活性比前段要高些。鲢的淀粉酶活性分布规律与上述情况相似,但鳙的淀粉酶活性在肠粘膜各段中由前向后呈递减趋势,并以第Ⅰ段为最大。在相同的器官组织中,鳙蛋白酶活性高于鲢,但差异不显著（Ｐ＞０．０５）,而淀粉酶活性则相反。这两种鱼的蛋白酶活性适宜ｐＨ为８．０～８．５,淀粉酶活性在ｐＨ７．５时最适。在鲢的肠内容物中发现有纤维素酶活性,但在肠粘膜中未检出该酶。     

Effects of polysaccharide from fermented wheat bran on growth performance,muscle composition,digestive enzyme activities and intestinal microbiota in juvenile common carp

The present study evaluated the effect of dietary polysaccharide from fermented wheat bran on growth performance, muscle composition, digestive enzyme activities and intestinal microbiota of juvenile common carp (Cyprinus carpio L.). Four diets supplemented with 0 (D0), 1.0 (D1), 2.0 (D2) and 4.0 (D3) g/kg fermented wheat bran polysaccharide (FWBP) were prepared. Fish were randomly assigned into four groups, each group having four replicates (30 fish/replicate) and fed with experimental diets for 8 weeks. Compared with the D0 group, the D3 group had higher final body weight and weight gain (p < .05). The amylase activity in the intestine and hepatopancreas of D1 and D2 groups and pepsin and trypsin activities in the hepatopancreas of D2 and D3 groups were significantly higher than D0 group. Significantly increased muscle protein and lipid content while reduced moisture content was observed in D1 group; additionally, D3 group had significantly lower muscle lipid content. The population of Bifidobacterium spp. in the intestine digesta of D1 and D2 groups was significantly increased, whereas lactic acid bacteria (LAB) and Escherichia coli were unaffected. Collectively, dietary FWBP supplementation could enhance digestive enzyme activities and Bifidobacterium population and change the muscle composition and improve growth performance of common carp.     

鳜鱼、青鱼、草鱼、鲤、鲫、鲢消化酶活性的研究

鳜鱼、青鱼、草鱼、鲤、鲫、鲢肝脏蛋白酶活性低于肠蛋白酶。肠蛋白酶活性，鳜鱼最高，其余依次为青鱼、鲤、鲢、草鱼、鲫。鳜鱼、鲢的肠蛋白酶活性由前肠向后肠递减；而青鱼、鲤、草鱼的则由前肠向后肠递增；鲫则中肠活性最低。六种鱼不同组织的脂肪酶活性因鱼而异。青鱼、鳜鱼、鲤、鲢肝脏中脂肪酶活性低于肠脂肪酶活性（Ｐ＜０．０１）；而草鱼和鲫肝脏中脂肪酶活性与肠脂肪酶活性差异不明显（Ｐ＞０．０５）。六种鱼肝脏中脂肪酶活性由高到低依次为鳜鱼＞鲢＞鲫＞草鱼＞青鱼＞鲤；肠脂肪酶活性顺序为鳜鱼＞鲢＞青鱼＞鲤＞草鱼＞鲫。脂肪酶活性与鱼类食性无明显相关性。不同组织间淀粉酶活性存在差异。鳜鱼的肝脏和肠均有较高淀粉酶活性，青鱼和鲫肝脏中淀粉酶活性低于肠中的；草鱼、鲤和鲢肝脏中淀粉酶活性高于肠中的，但差异不显著。六种鱼中鳜鱼的肝脏淀粉酶活性明显高于其它五种无胃鱼，它们的淀粉酶活性顺序依次为鳜鱼＞鲢＞鲤＞草鱼＞青鱼＞鲫；肠淀粉酶活性顺序为鳜鱼＞鲫＞鲤＞青鱼＞鲢＞草鱼。     

Deficiency of dietary niacin decreases digestion and absorption capacities via declining the digestive and brush border enzyme activities and downregulating those enzyme gene transcription related to TOR pathway of the hepatopancreas and intestine in young grass carp (Ctenopharyngodon idella)

To investigate the effects of niacin on growth, digestion and absorption capacity, and the potential mechanism for digestive and brush border enzyme activities, grass carp (Ctenopharyngodon idella) (256 ± 0.41 g) were fed diets containing 3.95 (basal diet group), 14.92, 24.98, 35.03, 44.97 and 55.01 mg niacin kg^?1 diet for 8 weeks. Results indicated that percentage weight gain (PWG), feed intake and feed efficiency were the lowest in basal group (P < 0.05). Similarly, niacin deficiency decreased hepatopancreas trypsin, chymotrypsin, lipase and amylase activities (P < 0.05), intestinal Na⁺, K⁺‐ATPase, alkaline phosphatase, γ‐glutamyl transpeptidase and creatine kinase (CK) activities, the cholecystokinin (CCK) content in proximal intestine (PI) and growth hormone content in serum (P < 0.05). Furthermore, niacin deficiency downregulated gene expression of hepatopancreas trypsinogen 1, trypsinogen 2, chymotrypsinogen and amylase, intestinal Na⁺, K⁺‐ATPase alpha subunit isoform 1, Na⁺, K⁺‐ATPase alpha subunit isoform 8 and CK, and target of rapamycin (TOR) and S6 kinase 1 (S6K1) of hepatopancreas and intestine (P < 0.05), whereas upregulated eIF4E‐binding protein (4EBP) gene expression (P < 0.05). The niacin requirement for young grass carp (256–689 g) based on PWG, hepatopancreas trypsin activity and Na⁺, K⁺‐ATPase in PI was 34.01, 35.10 and 42.08 mg kg^?1 diet, respectively.     

鲤、鲢、鳙、草鱼消化道消化酶种类和活性的比较研究

采用聚丙烯酰胺凝胶电泳方法研究了鲤、鲢、鳙、草鱼亲鱼消化道消化酶的种类；用比色法及滴定法分别测其消化酶相对活性。结果表明：鲤、鲢、鳙、草鱼4种鱼消化道的蛋白酶、淀粉酶活性与食性有明显的相关性。其中，蛋白酶活性依次为鲤〉鲢〉鳙〉草鱼；淀粉酶活性依次为草鱼〉鲤〉鳙〉鲢。脂肪酶活性与食性的关系不明显。四种鱼消化酶的表达与消化道的组织结构特征相一致，消化道前2／5部位（G1、G2）的消化酶活性最高，是无胃鱼消化道的主要消化场所。消化道中央部位（G3）的消化酶活性次之，能将未消化完全的食糜进一步消化，而消化道后段（G4、G5）消化酶活性最低。可见，消化酶活性及消化功能呈递减形式，食物在消化道中随着消化、吸收，逐渐排出体外。本研究为鲤、鲢、草鱼、鳙4种鱼的营养生理学、高效饲料配比的研制及种质资源的科学管理提供理论依据。     

Enzyme producing bacterial flora isolated from fish digestive tracts

Isolationand enumeration of aerobic bacterial flora in the gastrointestinal tract of nineculturable freshwater teleosts, namely catla, rohu, mrigal, silver carp, grasscarp, common carp, tilapia, walking catfish and murrel have been carried out.Amylolytic, cellulolytic, lipolytic and proteolytic microflora were identifiedfrom the culture plate using selective media. The isolates were qualitativelyscreened on the basis of their extracellular enzyme producing ability. Theselected strains were further quantitatively assayed for amylase, cellulase,lipase and protease activities. Protease activity was exhibited by almost allthe bacterial isolates, while strains isolated from tilapia, grass carp andcommon carp showed considerable amylolytic and cellulolytic activities. Maximumactivity of lipase was exhibited by a strain isolated from silver carp. Thestudy indicates that there is a distinct microbial source of the digestiveenzymes – amylase, cellulase, lipase and protease, apart from endogenoussources in fish gut. The information generated from the present investigationmight contribute towards better feed formulations for carp at low cost,incorporating the enzyme producing bacterial isolates as probiotics.