Effect of tank proportions on survival of seven-band grouper Epinephelus septemfasciatus (Thunberg) and devil stinger Inimicus japonicus (Cuvier) larvae

We examined the effects of rearing‐tank proportions on early survival, surface death and growth of the seven‐band grouper Epinephelus septemfasciatus (Thunberg) and the devil stinger Inimicus japonicus (Cuvier). Fertilized eggs were introduced into three differently shaped 100 L rearing tanks. The three tanks had different water surface areas, and included a shallow tank (S; 71 × 26 cm in diameter and depth, respectively), an intermediate tank (I; 57 × 39 cm) and a deep tank (D; 44 × 70 cm). Both species showed their highest survival rate and the lowest numbers of surface death in the D tank (P<0.05). There were no significant differences between fish reared in the three tank shapes in notochord length, total length, growth rate and dry weight. Rearing‐tank shape affected larval movement in the water column, with the duration of larval movement under the water surface being the shortest in the D tank. These results suggest that using a rearing tank of a suitable shape could significantly reduce the surface death of marine fish larvae.     

Characterization of digestive enzyme activity during larval development of gilthead seabream (Sparus aurata)

The evolution of the digestive enzyme equipment in seabream from hatching to 30 days old larvae was studied; there was a progressive increase in the activity of protease, amylase and acid and alkaline phosphatase from day 15 onwards. The use of specific inhibitors, and SDS-PAGE provided evidence to suggest that most of the proteases belonged to the serine group. A high -amylase activity was also denoted. Zymograms of larval extracts indicated that exogenous food has more a qualitative than a quantitative role in the secretion of digestive enzymes in this species.     

厚颌鲂仔稚鱼消化酶活性变化研究

测定了厚颌鲂(Megalobrama pellegrini)仔稚鱼从出膜到30日龄的生长、可溶性蛋白含量及消化酶活性。结果表明:初孵仔鱼即能检测到胰蛋白酶、淀粉酶、脂肪酶活性,2日龄检测到碱性磷酸酶活性。投喂组可溶性蛋白含量随日龄的增加显著增加(P<0.05),饥饿组可溶性蛋白含量6日龄前增加不显著,6日龄后开始下降。所测消化酶的全活力与比活力呈现出不同的变化模式。各消化酶的全活力基本是随日龄的增加而增加。投喂组胰蛋白酶比活力在10日龄达最大值,随后显著下降;饥饿组胰蛋白酶比活力在4日龄达最大值,随后显著下降。投喂组淀粉酶在5日龄达到最大值,从6日龄到12日龄维持在一个较低水平,随后增加至15日龄的(0.912±0.090)mU/mg,在试验后期均维持在一个相对较低的水平;饥饿组淀粉酶比活力的最大值出现在1日龄,从2日龄到8日龄变化不显著,8日龄后显著下降。投喂组脂肪酶比活力在18日龄达最大值,随后显著下降;饥饿组脂肪酶比活力1日龄到6日龄变化不显著,6日龄后显著下降。投喂组碱性磷酸酶比活力从2日龄到12日龄显著上升,12日龄达到最大值,随后显著下降;饥饿组碱性磷酸酶比活力从2日龄到8日龄显著上升,8日龄达到最大值,随后显著下降。     

瓦氏黄颡鱼仔稚鱼发育过程中消化酶活性变化研究

测定了瓦氏黄颡鱼(Pelteobagrus vachelli)仔稚鱼从出膜至30日龄时的生长、可溶性蛋白含量及消化酶活性.结果表明,在仔鱼出膜后第1天,蛋白酶、淀粉酶、脂肪酶均能检测到活性,而胃蛋白酶活性在第22日龄才检测到.可溶性蛋白的含量随日龄的增加而增加.蛋白酶、淀粉酶、脂肪酶的活性随着仔稚鱼的发育而发生变化,其全活力与比活力呈现出不同的变化模式.蛋白酶比活力在仔鱼4日龄时达到最大值(4.193±0.04)U/mg protein;之后随日龄的增加比活力降低,直至15日龄达到最小值(0.452±0.07)U/mg protein.之后从15日龄到30日龄比活力逐渐增加.淀粉酶比活力从1日龄逐渐增加,至4日龄达到最大值(4.611±0.12)U/mg protein,从10日龄到18日龄维持在一较低水平,随后随日龄的增加淀粉酶比活力增加.脂肪酶比活力从1日龄到3日龄逐渐增加,到3日龄达到最大值(4.398±0.07)U/mgprotein,3日龄以后直到30日龄,脂肪酶比活力随日龄的增加而降低,并且一直处于一较低的水平.瓦氏黄颡鱼发育过程中,主要消化酶活性随生长变化显著,反映瓦氏黄颡鱼随着生长其消化功能逐渐完善.     

鬼鲉早期发育阶段的摄食节律与饥饿致死时间

采用实验生态学方法研究鬼鲉(Inimicus japonicus)早期发育阶段的摄食特性和仔鱼饥饿致死时间.结果表明,仔鱼和营游泳生活的稚鱼摄食高峰出现在白天,并以白天摄食为主,6日龄仔鱼、17日龄营游泳生活稚鱼的摄食率高峰为10:00和14:00.随着生活习性的转变,其摄食节律也发生明显变化,稚鱼和幼鱼在营底栖生活方式的阶段,以夜间摄食为主,另一摄食小高峰出现在早晨.进入底栖生活阶段的29日龄稚鱼、40日龄幼鱼的摄食高峰为22:00和6:00.鬼鲉仔、稚、幼鱼各阶段的日摄食率分别为:6日龄仔鱼69.7%,17日龄稚鱼57.7%,29日龄稚鱼42.3%,40日龄幼鱼17.7%.在饥饿条件下鬼鲉初孵仔鱼、1、3、5、8和10日龄仔鱼的全部死亡时间分别为192 h、216 h、168 h、80 h、178 h和264 h,半数死亡时间分别为144h、120 h、120h、60h、108 h和144h,说明5日龄仔鱼是鬼鲉早期发育中最为敏感的阶段.饥饿仔鱼呈现头大、体短、身瘦、体色发白等形态特征,行为上表现为游动缓慢,反应迟钝和静卧底部.长期饥饿后仔鱼脑后部下陷明显,其形态特征可作为判断仔鱼饥饿致死的重要指标.     

野生与养殖哲罗鱼消化系统及消化酶的比较研究

采用组织学方法对野生与养殖哲罗鱼（Hucho taimen Pallas）消化系统的形态及组织结构进行比较研究,采用聚丙烯酰胺凝胶电泳方法比较了两个群体哲罗鱼3种消化酶的相对含量。结果表明,野生与养殖哲罗鱼的消化系统在形态上存在一定差异,在组织结构上无明显差异。两个群体哲罗鱼在食道与胃酯酶相对含量上差异不显著（P〉0．05）,在肝脏与幽门盲囊酯酶相对含量上差异显著（P〈0．05）,在肠道酯酶的相对含量上差异极为显著（P〈0．01）。两个群体哲罗鱼在食道与胃淀粉酶的相对含量上差异不显著（P〉0．05）,在肝脏与肠道淀粉酶的相对含量上差异显著（P〈0．05）,在幽门盲囊淀粉酶的相对含量上差异极为显著（P〈0．01）。哲罗鱼的蛋白酶表达存在一定的pH依赖性,在体外酸性条件下（pH3）,两个群体哲罗鱼只有食道与胃有蛋白酶表达;在体外碱性条件下（pH9）,两个群体哲罗鱼只有幽门盲囊与肠道有蛋白酶表达;两种pH条件下,两个群体哲罗鱼各组织中蛋白酶的相对含量差异均不显著（P〉0．05）。两个群体哲罗鱼的消化系统结构及消化酶含量所表现出的差异与其生活环境及食物有关。[中国水产科学,2008,15（2）：330-336]     

Trypsin enzyme activity during larval development of Litopenaeus vannamei (Boone) fed on live feeds

Larval stages of the Pacific white shrimp, Litopenaeus vannamei (Boone) were fed standard live diets of mixed microalgae from the first to the third protozoea (PZ1 to PZ3), followed by Artemia nauplii until post‐larvae 1 (PL1). Trypsin enzyme activity for each larval stage was determined using N‐α‐p‐toluenesulphonyl‐l ‐arginine methyl ester (TAME) as a substrate. Results were expressed as enzyme content to assess ontogenetic changes during larval development. Tissue trypsin content (IU µg^?1 DW for each larval stage) was significantly highest at the PZ1 stage and declined through subsequent stages to PL1. This contrasts with previously observed patterns of trypsin development in Litopenaeus setiferus (Linnaeus) and other penaeid genera, which exhibit a peak in trypsin activity at the third protozoea/first mysis (PZ3/M1) larval stage. Litopenaeus vannamei larvae transferred to a diet of Artemia at the beginning of the second protozoea (PZ2) stage were significantly heavier on reaching the first mysis stage (M1) than those fed algae, while survival was not significantly different between treatments. At both PZ2 and PZ3 stages, trypsin content in larvae feeding on Artemia was significantly lower than in those feeding on algae. The rapid decline in trypsin content from PZ1 and the flexible enzyme response from PZ2 suggest that L. vannamei is physiologically adapted to transfer to a more carnivorous diet during the mid‐protozoeal stages.     

罗氏沼虾幼体及成虾消化酶活性

罗氏沼虾幼体及成虾消化酶活性魏华，赵维信（上海水产大学，上海２０００９０）关键词罗氏沼虾，幼体，消化酶ＴＨＥＡＣＴＩＶＩＴＩＥＳＯＦＤＩＧＥＳＴＩＶＥＥＮＺＹＭＥＳＯＦＬＡＲＶＡＥＡＮＤＡＤＵＬＴＩＮＭＡＣＲＯＢＲＡＣＨＩＵＭＲＯＳＥＮＢＥＲＧＩＩ￥...     

泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化

研究了泥鳅(Misgurnus anguillicaudatus)从孵化至30 DAH(日龄,Days after hatching)几种消化酶及碱性磷酸酶比活力的变化情况。胃蛋白酶直至30 DAH仍未检出活性。而胰蛋白酶表现出较高的比活力,其比活力在初次摄食之后显著上升,6 DAH达到最大值之后开始显著降低(P<0.05)。脂肪酶与淀粉酶的变化模式相似,在内源性营养向外源性营养转变及仔鱼向稚鱼转变这两个时间段出现两个高峰值。碱性磷酸酶比活力在2-6 DAH显著上升(P<0.05),之后开始下降并趋于平稳。研究表明,泥鳅在仔稚鱼阶段只具有结构性的胃而缺乏分泌细胞的分化。2-6 DAH是泥鳅仔鱼肠道功能迅速发育的阶段,也是向成鱼消化模式转变的一个重要过程。脂肪酶和淀粉酶比活力的持续性表明了泥鳅仔鱼对糖类和脂肪有较好的利用能力。     

Characterization of chymotrypsin activity during early ontogeny of larval red drum (Sciaenops ocellatus)

The temporal evolution of chymotrypsin activity during early ontogeny of laboratory reared red drum larvae was accomplished using a combination of biochemical assays and electrophoretic methods (substrate SDS-PAGE). Optimal functional conditions for chymotrypsin were also determined. Chymotrypsin activity was first detected prior to the onset of exogenous feeding. Total chymotrypsin activity increased with age and standard length. Specific activity was greatest on day 10 post-hatch. Maximal chymotrypsin activity was observed at 50 °C, pH 7.8, and Ca²⁺ concentration of 25 mM. Using substrate gel electrophoresis and specific inhibitors the molecular weight of red drum chymotrypsin was estimated to be 26–27 kD. Our results indicate that the digestive system of red drum larvae is capable of alkaline proteolysis before first feeding and suggest that chymotrypsin may have potential as an indicator of nutritional condition.     

Identification and partial characterization of selected proteolytic enzymes in the digestive system of giant freshwater Prawn Macrobrachium rosenbergii (De Man) Postlarvae

Biochemical assays and substrate SDS-PAGE were conducted to partially characterize and identify various types of proteases present in the digestive tract of PL₁₅ giant freshwater prawn ( Macrobrachium rosenbergii ). Casein hydrolytic assay of the enzyme extracts showed major proteolytic activities at pH 3.0, 6.0 and 9.0, while assay of preincubated enzyme extracts with phenylmethylsulphonyl fluoride (PMSF), a serine protease inhibitor produced a 33.17% reduction in alkaline protease activity. When specific inhibitors tosyl-lysine chloromethyl ketone and tosyl-phenylalanine chloromethyl ketone were used, they resulted in a reduction in activity of proteases in the enzyme extracts by 82.41% and 55.03%, respectively, confirming the presence of trypsin and chymotrypsin, while ethylenediamine tetraacetic acid produced protease activity reduction in 33.92% showing the presence of metalloproteases in the digestive tract of the prawn. Further characterization of the alkaline proteases using SDS-PAGE technique, after incubating the extract in the presence or absence of specific inhibitors, produced six bands corresponding to molecular masses of between 13.48 and 136.1 kDa; two trypsin bands of 13.48 and 36.4 kDa, three chymotrypsin bands in the range of 23.0–73.4 kDa and one for metalloprotease of 136.1 kDa, all of which were identified from a zymogram. This study suggests that protein digestion in M. rosenbergii is initiated by an acid protease followed by a combination of action of alkaline proteases: trypsin, chymotrypsin and metalloproteases.     

Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus (Bloch 1790)

Ontogenetic development of digestive system in crimson snapper (Lutjanus erythopterus Bloch 1790) larvae was histologically and enzymatically (alkaline phosphatase, amylase, lipase, pepsin, trypsin) examined from hatching to 36 days post hatching (DPH). The ontogenetic development of crimson snapper larval fish ontogeny was divided into three distinct phases: phase I starting from hatch to the onset of exogenous feeding, phase II starting from first feeding (2–3 DPH) until the formation of gastric glands (13–14 DPH) and phase III beginning from the appearance of gastric glands and continuing onwards. The specific activities of amylase, lipase, trypsin showed sharp increase and reached to the maximum from hatch to 4 DPH, 10 DPH, 20 DPH, respectively, followed by a declining trend with irregular fluctuation. In contrast to other enzymes, the specific activities of alkaline phosphatase showed a gradual increase from hatch to 29 DPH, followed by a sharp increase towards 36 DPH. The specific activity of pepsin was firstly detected on 17 DPH and gradually increased towards the end of this study. The total activities of these five enzymes showed a gradual increase till 29 DPH, followed by a sharp increase towards 36 DPH except for amylase and lipase reaching maximum at 32 DPH. The present study provides better understanding of the digestive ontogeny of crimson snapper during the larval stage and a guide to feeding and weaning of this economically important fish in hatcheries.     

Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus

The growth, survival and trypsin, lipase and amylase activities of red drum larvae were measured in two experiments. For the first trial, a group was fed live prey only (L) and another group was fed a combination of a microparticulate diet (MPD) and live food (L-MP). For the second growth trial a group fed the MPD only (MP) and a starvation group (ST) were examined in addition to the L and L-MP treatments. Enzyme activities of live prey were measured to estimate their possible contribution to larval digestion. No significant ( P  > 0.05) differences in final size and survival were observed between treatments L and L-MP. Larvae subjected to starvation or fed the MPD diet alone were smaller than treatments fed live prey and did not survive past days 5 and 14, respectively. Trypsin, lipase and amylase activities were detectable at hatching. No significant differences ( P  > 0.05) in total enzyme activities among treatments were observed before day 14. Specific activity of trypsin, lipase and amylase peaked on day 3 (prior to first feeding) and subsequently decreased. For trypsin, the percentage of enzyme activity potentially attributable to ingested prey increased with age to a maximum of 17%. For lipase and amylase this fraction was less than 5% throughout the study, except on day 8 (12% and 24%, respectively). The lack of significant differences observed in the activity of digestive enzymes among treatments suggests that dietary regime, availability of prey and possible effects of exogenous enzymes did not significantly influence enzyme activity. Therefore, the lower growth rate observed in the L-MP, MP and starved treatments cannot be attributed to low digestive enzyme production of the enzymes measured. It is more likely that the MPD failed to supply the required nutrients for adequate development.     

Digestive enzyme activity during larval development of the Senegal sole (Solea senegalensis)

The activities of the main digestive enzymes (proteases, amylase, lipase) as well as those of acid and alkaline phosphatases were assessed during the larval development of the Senegal sole, Solea senegalensis. Important variations in specific activities of all the enzymes were observed during the period of study and were mostly related to the beginning or the end of metamorphosis. Both acid and alkaline protease activities were identified at early stages of development. Acid proteases accounted for only 10% of total protease activity in a 9 days-old larva, but exceeded 75% in a 33 days-old individual. Maximum lipase activity was related to the development of exocrine pancreas (days 6 to 10) and to metamorphosis. It is suggested this can be related to the metabolism of lipid reserves taking place during this morphological change. Activity of alkaline phosphatase decreased from day 5 to day 20 but therafter revealed a sharp increase, possibly linked to the development of enterocytes. The pattern of development of the main digestive enzymes found in S. senegalensis is similar to that described in other flatfish species.     

盐度对奥尼罗非鱼仔稚鱼生长、存活及其消化酶活力的影响

研究了不同盐度对奥尼罗非鱼仔、稚鱼生长、存活和消化酶活力的影响。试验设计2、4、6、8、10和纯净淡水0（对照组）共6个盐度梯度组,仔鱼的初始体重0.0080.001 g。结果表明,不同盐度对奥尼罗非鱼仔、稚鱼生长有显著差异（P0.05）,盐度4和6时,仔、稚鱼的生长速度和绝对增重率最高,分别为0.08 cmd-1和0.033 gd-1;盐度10时较低,只有0.06 cmd-1和0.023 gd-1;仔、稚鱼的肥满度介于3.43~3.81,各组之间差异不显著（P0.05）;盐度0~10时,各组存活率介于92.3%~94.7%,幼鱼所占比例介于66.0%~67.3%,各组之间差异不显著（P0.05）;胃蛋白酶和胰蛋白酶活力随盐度升高而降低,均在盐度0时活力最高,10时最低;脂肪酶和淀粉酶活力在盐度4时达到最高,10时活力最低。根据试验结果,奥尼罗非鱼仔、稚鱼培育的盐度最好控制在4.6左右。     

银鲳仔鱼消化系统的组织学研究

对出膜后1~12 d银鲳(Pampus argenteus)仔鱼的消化系统进行了形态学和组织学观察。在水温为22~24℃和盐度25~28情况下,初孵仔鱼具很大的卵黄囊,消化管为一简单的直形盲管,管腔狭窄,口和肛门尚未与外界接通。3 d仔鱼消化系统分化加快,在卵黄囊凹陷部位出现2~3个弯曲,已初步分化出食道、胃、肠和肝脏,肠管也变粗。4 d仔鱼消化系统各器官初步形成。5 d仔鱼出现侧囊,并见部分卵黄囊和油球。7 d卵黄囊和油球基本被完全吸收,仔鱼主动摄食轮虫和小球藻,从内源性营养向外源性营养的过渡基本完成。12 d以后仔鱼肝脏明显分为两叶,体积增大;幽门盲囊指状分支已增加到几十根;食道、胃和肠的黏膜皱褶明显增多和加深,肠黏膜上皮细胞高度增加,游离面纹状缘发达;胃黏膜的单层柱状上皮高度和胃腺细胞数量也明显增加;但整个消化道的黏膜下层、肌肉层均不发达,说明12 d仔鱼已具初步消化和吸收功能     

灰鲳仔稚鱼消化系统的发育

对出膜后1~35 d灰鲳仔稚鱼的消化系统进行了形态学和组织学观察。初孵仔鱼具有很大的卵黄囊,消化管为一简单的直形盲管。2日龄仔鱼消化系统中肠区首先开始分化,内面可见纹状缘。3日龄仔鱼胃区膨大,卵黄消失,肠道分化出小肠和直肠。侧囊在4日龄仔鱼时出现分化。5日龄仔鱼观察到幽门盲囊的初始结构,油球消失,从内源性营养向外源性营养过渡基本完成。6日龄仔鱼摄食主动。14日龄稚鱼肝脏分为两叶,体积增大,胰脏体积增大,胃弯曲呈"U"形,幽门盲囊指状分支已增加到几十个。22日龄稚鱼肠道弯曲更多,在胸腹腔中肠道盘曲程度已接近幼鱼。28日龄稚鱼侧囊背腹形成一纵隔,出现角质剌。35日龄稚鱼胃腺已十分丰富,侧囊结构也已完善,标志着灰鲳消化系统更趋完善。     

Ontogeny of digestive and immune system organs of larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory

ABSTRACT:   A histological examination was made of the ontogenetic development of the digestive and immune systems of the larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory. The liver, gall bladder, pancreas and the demarcating region between the intestines and rectum were formed within 3 days post-hatch (dph). During the preflexion phase (within 16 dph), revolution of the intestine concluded, and pharyngeal teeth and the mucous cells of the esophagus were differentiated. In the transitional period to the juvenile stage (25 dph), the blind sac of the stomach, gastric glands and pyloric caeca began to form. From the viewpoint of the differentiation phase of the adult-type digestive system, the kelp grouper is similar to Heterosomata, hitherto reported. The primordial thymus, kidney and spleen were present at 12, 1 and 6 dph, and the small lymphocytes in these lymphoid organs appeared at 21, 30 and 33 dph, respectively. The developmental sequence of the lymphoid organs and the appearance ages of the lymphoid organs and small lymphocytes in the lymphoid organs in the kelp grouper are similar to those of other marine fish previously reported, except for the Pacific bluefin tuna Thunnus orientalis .     

鲇仔、稚鱼消化系统胚后发育的组织学观察研究

通过石蜡包埋切片法对鲇(SilurusasotusL.)仔、稚鱼消化系统胚后发育进行了较系统的组织切片观察。本研究描述了全长5.0～22.5mm的鲇摄食器官、消化器官胚后发育的组织学结构特征。观察发现,1～3日龄为内源性营养阶段,卵黄囊很大,2日龄消化道出现裂缝状腔隙,3日龄基本贯通但未开始摄食;4～6日龄为混合营养阶段,卵黄囊被逐步吸收,主要靠吞食轮虫、小型枝角类等为食;6日龄以后卵黄囊消失,进入外源性营养阶段,捕食能力增强。观察还发现,鲇前咽顶壁始终平直无粘膜皱褶;颌齿和咽齿为斜生尖锥状的同型齿,数量多、排列紧密,与相应的骨骼牢固地骨性固着;后咽、食道的粘膜上皮内粘液细胞极多,深层结缔组织肌肉层发达。这些构造适应于鲇的完全吞食摄食方式。胃腔小,前肠膨大、中肠粘膜上皮细胞纹状缘发达,肝脏和胰脏发育速度较快。胃的消化功能较弱,主要储存、消化场所为前肠,吸收场所在中肠。鱼苗4～6日龄下塘适宜,6～8日龄可开始诱其摄食人工饲料。     

Evaluation of weaning performance of California halibut (Paralichthys californicus) larvae using growth,survival and digestive proteolytic activity

This study evaluated weaning success of California halibut, Paralichthys californicus, larvae onto a microdiet at various stages of development utilizing growth, survival and digestive enzyme activity. Weaning onto a microdiet was evaluated at 16, 26, 36 and 46 days posthatch (dph). Alkaline and acid proteases and leucine aminopeptidase activities were measured after weaning and compared between the weaned treatment and Artemia‐fed controls. Survival was significantly lower in the microdiet‐fed treatments compared to the control groups. Growth was significantly reduced in all weaning treatments compared to the control, except for the 46 dph group. No differences in enzyme activities were detected between treatment and diet at 16 and 26 dph; however, activities were higher for the microdiet‐fed larvae at 36 and 46 dph. This study demonstrates that California halibut larvae possess a differentiated and effective digestive system early in development and can be weaned with relative success (>40% survival) before completion of the metamorphosis (i.e., 36 dph). The lack of weaning success at an early date cannot be entirely because of the absence of a functional stomach but could be related to, among other factors, the low‐microdiet ingestion rates observed and higher leaching of smaller microdiets.