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1.
The appearance of functionally developed gastric glands is commonly considered as the transition from the larval to the juvenile stage in fish, since it means the switch from the less efficient alkaline digestion to a more efficient acid digestion characteristic of adult specimens. From that moment, fish are supposedly able to better assimilate nutrients from inert diets. Acid digestion takes place by the action of pepsin activity and hydrochloric acid, both secreted by the gastric glands of the stomach. Pepsinogen is the precursor of pepsin which is converted into active enzyme by the action of hydrochloric acid secreted by the proton pump. The goal of this work was to asses the ontogeny of pepsinogen and gastric proton pump expression along larval development of red porgy using RT-PCR and in situ hybridization techniques. Firstly, red porgy specific pepsinogen and proton pump partial sequences were isolated. Amplification products presented 615 and 591 bp and were identified as pepsinogen IIb and the -subunit of the proton pump (H+/K+-ATPase) by sequencing, respectively. Both sequences were aligned to several predicted pepsinogen and proton pump polypeptides from different vertebrate species showing elevated homologies with them, especially in the case of the proton pump, the identity of which was never less than 90%. Pepsinogen and proton pump expressions were detected from 30 days post-hatching (dph), increasing with development. Proton pump expression was localized in the gastric glands of red porgy larvae as revealed by in situ hybridization, showing increasing signal intensity along the digestive system development. Such results indicated that at 30 dph red porgy starts to acquire the adult digestive capacity and therefore inert diets should be better utilized from that time onwards.  相似文献   

2.
鳜胃蛋白酶原基因cDNA全长的克隆与序列分析   总被引:6,自引:2,他引:4  
吴雪峰  赵金良 《水产学报》2008,32(6):971-976
利用RT–PCR和cDNA末端快速扩增法(RACE)克隆鳜(Siniperca chuatsi)胃蛋白酶原基因cDNA全长序列,并对该基因的结构特征和系统进化关系进行了分析。鳜胃蛋白酶原基因cDNA序列全长1367 bp,5′端非翻译区43 bp,3′端非翻译区187 bp,开放阅读框(ORF)1137 bp,共编码378个氨基酸。鳜胃蛋白酶原氨基末端存在信号肽和激活肽序列,序列中含有催化活性必需的2个天冬氨酸残基和构成二硫键的6个半胱氨酸残基。鳜胃蛋白酶原氨基酸序列与其他脊椎动物胃蛋白酶原氨基酸序列的同源性为59.9%~91.2%,表明胃蛋白酶原基因在脊椎动物的长期进化中比较保守。鳜胃蛋白酶原基因的成功克隆不仅为进一步研究该基因的时空表达奠定基础,而且为鱼类胃蛋白酶原的分子特征和进化提供了新的资料。  相似文献   

3.
The major digestive enzymes in Pacific bluefin tuna Thunnus orientalis larvae were characterized, and the physiological characteristics of the enzymes during early ontogeny were clarified using biochemical and molecular approaches. The maximum activity of trypsin (Try), chymotrypsin (Ct) and amylase (Amy) was observed at pH 6–11, 8–11 and 6–9, respectively. Maximum activity of Try, Ct and Amy occurred at 50 °C, that of lipase (Lip) was at 60 °C and that of pepsin (Pep) was at 40–50 °C. These pH and thermal profiles were similar to those for other fish species but differed from those previously reported for adult bluefin tuna. Enzyme activity for all enzymes assayed was found to decrease at high temperatures (Try, Ct, Amy and Pep: 50 °C; Lip: 40 °C), which is similar to findings for other fish species with one marked exception—increased Try activity was observed at 40 °C. Lip activity appeared to be dependent on bile salts under our assay conditions, resulting in a significant increase in activity in the presence of bile salts. Ontogenetic changes in pancreatic digestive enzymes showed similar gene expression patterns to those of other fish species, whereas marked temporal increases in enzyme activities were observed at 10–12 days post hatching (dph), coinciding with previously reported timing of the development of the pyloric caeca in bluefin tuna larvae. However, complete development of digestive function was indicated by the high pep gene expression from 19 dph, which contradicts the profile of Pep activity and previously reported development timing of the gastric gland. These findings contribute to the general knowledge of bluefin tuna larval digestive system development.  相似文献   

4.
The pancreatic digestive enzymes, trypsin, chymotrypsin, lipase and amylase were partially characterized, and changes in their activities were examined during the initial ontogeny of Japanese eel Anguilla japonica larvae from 5 to 34 days post-hatching (dph). The pH optima of the eel larval enzymes were narrower than those other fish species; trypsin activity was highest at pH 9, chymotrypsin and amylase activities were highest at pH 7 and 8, and lipase activity was highest at pH 8 and 9. In an analysis of thermal profiles, the larval pancreatic enzymes had a high optimal temperature and high thermal stability, which are typical of fish from the tropics. At 12 and 13 dph, lipase activity and gene expression levels of trypsin (-a and -b), lipase and amylase decreased markedly, suggesting a marked change in larval metabolism at that time. These data could be useful in the development of artificial larval diets in Japanese eel.  相似文献   

5.
Pepsinogens are precursors of pepsins, which are gastric digestive proteinases that degrade food proteins into peptides. In the study reported here, the cDNA and its corresponding genomic DNA of the golden mandarin fish (Siniperca scherzeri, Perciformes) pepsinogen C (PGC) were cloned and sequenced. The golden mandarin fish PGC gene was deduced to have nine exons and eight introns, a structure similar to the PGCs of other vertebrates. The full-length cDNA was found to contain a 37-bp 5′-untranslated region, a 1,164-bp open reading frame, and a 304-bp 3′-untranslated region and the PGC protein to consist of a signal peptide, an activation segment, and a pepsin moiety. A sequence analysis revealed that pairwise sequence similarities of PGC proteins are around 70% between golden mandarin fish and other vertebrate groups, and around 90% within the fish group. A comparison of vertebrate PGC protein sequences revealed two motifs. One was in the activation segment that occurred only in the mammal and avian PGCs, suggesting that PGCs active in homeotherms (mammal and avian) have different activation mechanisms than those in poikilotherms (amphibian and fish). The second was in the pepsin moiety that occurred only in fish, suggesting the primitive position of fish among vertebrates. PGC mRNA is mainly expressed in the stomach and esophagus and at much lower levels in the skin and muscle. Taken together with results reported from other studies, the results reported here will lead to a better understanding of the molecular mechanisms of fish digestive physiology and the evolution of fish pepsinogen genes.  相似文献   

6.
The purpose of this experiment has been to evaluate the suitability of commercially available early-weaning microdiets (MDs) for the production of sea bream early juveniles and in comparison with late-weaning protocols already in use by Hellenic marine fish hatcheries. Four sea bream experimental groups were allocated in rearing tanks of a commercial Hellenic marine fish hatchery. Each group represented a different protocol (A, B, CA, and CB) based on the combination of two different early-weaning MDs (A and B) and a late-weaning diet (C). In addition, the late-weaning protocols have received Artemia instar II only and not Artemia instar I. In protocol A, Artemia instar I first feeding and the early-weaning diet A were administered at 17 days post-hatch (17 dph). In protocol B, Artemia instar I first feeding started at 15 dph and the early-weaning diet B was administered at 18 dph. In the C protocols, Artemia instar II first feeding started at 20 dph and the early-weaning diets (A or B) were administered at 25 dph. All protocols have received the late-weaning diet C only after the 50th dph. The experiment lasted for 65 days. By the end of the experiment, early-weaning protocol A and late-weaning protocol CB had similar wet weights, but still lower to the wet weights recorded for the late-weaning protocol CA. These results cannot be explained solely by the nutritional profile of each weaning diet. The larval fatty acid profile of each protocol and at various time intervals reveals the importance that the succession of Artemia, rotifers, and MDs has for each protocol and not just the nutritional profile of the weaning diets, per se.  相似文献   

7.
The impact of increased incorporation of plant ingredients on diets for rainbow trout was evaluated in terms of gene expression of gastric (gastric lipase, pepsinogen) and intestinal (prolidase, maltase, phospholipase A2) digestive enzymes and nutrient transporters (peptide and glucose transporters), as well as of postprandial levels of plasma glucose, triglycerides and total free amino acids. For that purpose, trout alevins were fed from the start of exogenous feeding one of three different experimental diets: a diet rich in fish meal and fish oil (FM–FO), a plant-based diet (noFM–noFO) totally free from fish meal and fish oil, but containing plant ingredients and a Mixed diet (Mixed) intermediate between the FM–FO and noFM–noFO diets. After 16 months of rearing, all fish were left unfed for 72 h and then given a single meal to satiation. Blood, stomach and anterior intestine were sampled before the meal and at 2, 6 and 12 h after this meal. The postprandial kinetics of gene expression of gastric and intestinal digestive enzymes and nutrient transporters were then followed in trout fed the FM–FO diet. The postprandial profiles showed that the expression of almost all genes studied was stimulated by the presence of nutrients in the digestive tract of trout, but the timing (appearance of peaks) varied between genes. Based on these data, we have focused on the molecular response to dietary factors in the stomach and the intestine at 6 and 12 h after feeding, respectively. The reduction in FM and FO levels of dietary incorporation induced a significant decrease in the gene expression of gastric lipase, GLUT2 and PEPT1. The plasma glucose and triglycerides levels were also reduced in trout fed the noFM–noFO diet. Consequently, the present study suggests a decrease in digestive capacities in trout fed a diet rich in plant ingredients.  相似文献   

8.
The ontogeny of the digestive tract was studied histologically in butter catfish Ompok bimaculatus from hatching to 30?days post-hatching (dph). At hatching, the digestive tract of butter catfish consisted of a straight tube with a smooth lumen dorsally attached to the yolk sac. Between 1 and 2 dph, the mouth opened, oral valves were visible and canine-like teeth and taste buds were detected. During this period, intestine was differentiated into the anterior and posterior intestine, and the digestive accessory glands were also developed. Exogenous feeding started at 2?dph, and there was a 2-day mixed endogenous–exogenous feeding period. Most of the yolk sac reserves were consumed between 2 and 3 dph, and by 5?dph, the yolk sac was completely depleted and no longer visible in histological sections. Between 3 and 4 dph, several vacuoles (neutral lipids) were observed in the intestine and also in hepatocytes, indicating a functional absorption of nutrients from food. At 8 dph, differentiation of gastric glands was noticed, and by 9–11?dph, there were abundant gastric tubular glands arranged along numerous longitudinal folds. During the same period, pyloric sphincter appeared as an epithelial fold that separated the stomach from the anterior intestine. From 12 dph to the end of the study at 30?dph, no noticeable histological modifications were observed. The development of gastric glands is considered as the last major events in digestive tract development and their presence designates the end of larval period and the onset of the juvenile period. Hence, it is suggested that, butter catfish larvae have a morphologically complete digestive tract by 12?dph. These findings on the development of the digestive system in butter catfish may lead to a better understanding of the ontogeny and would be useful to improve the larval rearing techniques of this promising catfish species for freshwater aquaculture diversification.  相似文献   

9.
Devil stinger is a valuable demersal scorpaenid fish while the rearing of stinger larvae still relies on live prey. This study was conducted to illustrate the development of the main digestive enzymes and digestive system during larval development of this species to provide evidence for the application of artificial feeds. Enzymatic and histological assays were conducted from 1 day post hatching (dph) to 36 dph in larvae. The result showed that the selected digestive enzyme activities increased significantly after 15 dph. Specifically, the total trypsin activities increased significantly from 18 dph to 33 dph. The total pepsin and amylase activities increased significantly first and thereafter decreased significantly. The lipase activities followed the similar pattern with trypsin. With regard to the histological study, the stinger larvae open their mouth to first feeding at 3 dph and turned into totally exogenous nutritional stage at 6 dph. In addition, mucous membrane, rich in goblet cells, was widely distributed in oesophagus epithelium at 18 dph. The height and amounts of gastric gland in cardia and main body of the stomach increased gradually with the development of stinger larvae after 15 dph. The intestine length of stinger larvae was short, and goblet cell was abundant in anterior intestine after 12 dph, not the posterior intestine. The ontogeny of liver and pancreas started from newly hatched stage, and the differentiation of liver was prior to pancreas. The above findings would provide evidence for the use of artificial feeds from the larval stage of stinger larvae (at least from 21 dph).  相似文献   

10.
Histomorphological changes in digestive tract of golden mahseer (Tor putitora) were examined in larvae [starting from hatching to 45 days post-hatching (dph)], fry, fingerling, and adult. Digestive tract appeared during hatching, on the dorsal side of yolk sac, as a straight tube with a narrow lumen. Mouth opening and appearance of liver and pancreas were observed at 2 dph, and subsequently anal opening, appearance of goblet cells in esophagus, and posterior intestine were evident at 3 dph. The remodeling of oral cavity in terms of epithelial stratification, appearance of taste buds, and goblet cells were observed in a window of 4–5 dph. Intestinal folding was found to be initiated at 8 dph. From 12 to 45 dph, thickening of oral and esophageal mucosal/extramucosal layers, increase in intestinal folding, increases in the density of goblet cells in entire gut were observed. Within the same time window, other histological changes such as disappearance of vacuoles in liver, and abundance of zymogen granules in pancreas were also observed. Supranuclear vesicles in mid-to-posterior intestine were found to be prominent from first feeding to 45 dph; however, this phenomenon was no longer evident in fry and fingerling. Overall, the increase in intestinal folding and complexity of extramucosal layer were found to be continuous from the first appearance to adult, and this inferred the fact that the nutritional physiology, in terms of digestion and assimilation, progressively changes throughout the life stages of golden mahseer. Findings of this study will, therefore, help in preparing diets for different life stages of this fish, and in addition, the present information widens the understanding of digestive physiology of golden mahseer.  相似文献   

11.
通过形态学与连续组织切片的方法,对美洲黑石斑鱼(Centropristis striata) 1-34 d仔鱼消化系统的胚后发育进行系统观察,分析描述鱼体消化道(食道、胃与肠道)以及消化腺(肝脏与胰腺)的发育过程.对1-15 d仔鱼连续取样,每次取样30尾,15 d后隔天取样,每次取样15尾.结果显示,在水温为(24±1)℃、盐度为30-32的条件下,初孵仔鱼卵黄囊体积很大,消化管为封闭的管状结构.美洲黑石斑鱼孵出3d后,口裂形成、开始摄食,肛门与外界连通,消化道逐渐分化形成食道、胃及肠道,肝脏、胆囊和胰腺也逐渐形成.7d后,卵黄囊与油球基本消失,食道、胃部以及肠道黏膜褶皱开始形成,消化道黏膜上皮细胞逐渐分化,肝脏出现脂肪颗粒,仔鱼具备了基本的摄食能力.11d时,仔鱼食道可见黏液细胞,随日龄的增加上皮组织中黏液细胞数量迅速增多,褶皱日益丰富;胃部分化形成贲门部、胃本体与幽门部,胃壁褶皱不断增多、伸长;肝脏血窦与中央静脉明显.20d时,鱼体胃腺形成,说明胃部消化、吸收蛋白质的能力增强;肠道次级黏膜褶皱出现,肠圈与褶皱更加复杂化;胰脏分布有大量酶原颗粒.32 d时,仔鱼消化道组织结构分明,自腔面向内依次为黏膜层、黏膜下层、肌层与浆膜层,消化道与消化腺结构和功能逐步完善.仔鱼3-7 d为内源性营养向外源性营养过渡期,应及时提供充足适口的生物饵料,仔鱼20 d后可以逐渐驯化投喂微型配合饲料.  相似文献   

12.
13.
With the development of successful aquaculture techniques, the Pacific bluefin tuna (PBT) has become the most important fish species in Japan. Early muscle growth and cellularity of muscle fibers have a profound effect on ultimate body size and meat quality of fish. Larvae and juveniles of full-cycle cultured PBT were sampled at 3, 15, 29, 41, 70, 128 and 218 days post hatch (dph). Transverse body sections until 41 dph and muscle sections from 41 dph thereafter from dorso-cranial or caudal areas were stained for muscle fiber morphological and morphometric analysis and muscle sections from the dorso-cranial region were used for ultra-structural observation. Red muscle fibers appeared by 15 dph at the horizontal septum both in epaxial and hypaxial regions. Other than a single layer of superficial red fibers and those at the horizontal septum, existence of red muscle fibers was apparent in the PBT myotome. The muscle fiber diameters varied in their size even in adjacent myotomes of PBT. Overall, growth throughout the various stages was a result of both hyperplasia and hypertrophy of muscle fibers. Muscle fiber diameters were greater in the dorso-cranial than in dorso-caudal region at 41, 70 and 128 dph. These results provide additional characterization of PBT growth, which may assist with managing fish production efficiency, harvest age and eating quality.  相似文献   

14.
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 .  相似文献   

15.
Turbot specimens were kept at three temperatures (T s ): warm (W) (21–22 °C), ambient (A) (17–18 °C) and cold (C) (13–14 °C) during the larval and early postlarval stages. At 90 days posthatching (dph), all of them were transferred to ambient T until 190 dph. At 2–3 dph, the specimens showed a monolayer of red muscle and immature white fibres; external or dermomyotome cells (presumptive myogenic cells) were observed on the surface of the red muscle. In the following stages, many myogenic cells and presumptive myogenic precursors were observed within the myotome, presumably derived of the dermomyotome. When comparing the growth at the same age (2, 10, 25, 37 dph), the body length and the muscle growth were positively influenced by the warm T, being the hyperplasia the muscle parameter more significantly influenced. The development rate was also positively correlated with the high T: the beginning of the metamorphosis took place at 15, 23 and 25 dph at W, A and C temperatures, respectively, with the highest body length values at ambient temperature. The metamorphosis finished at 25, 30 and 37 dph at W, A and C temperatures, respectively, with the highest body length values at warm temperature. However, the muscle cellularity was similar in all the groups at the end of the metamorphosis. At 90 and 190 dph, the largest body length was observed at W temperature. However, the muscle cellularity was similar between A and W; the number of fibres was similar in all the groups at 190 dph, which shows the beginning of a compensatory muscle growth in A and C, mainly in A.  相似文献   

16.
The aims of the present study are to determine whether triiodothyronine (T3) and/or thyroxine (T4) in tilapia larvae is gifted through the mother, and to investigate the change profiles of thyrotropin (TSH), thyroid follicular cells and type I deiodinase (D1) gene expression following larval development. T3 and T4 contents were measured using radioimmunoassay, thyrotropin was observed using immunocytochemistry, and the D1 gene was cloned and measured using real-time PCR. Results indicated that the β-TSH-immunoreactive cells (thyrotropin ICC) signals were detected at 9 dph (i.e., 9 days of post-hatching). Thyroid follicular cells were observed first at 3 dph, while the T3 contents of the whole body gradually decreased before 11 dph. T4 contents were detected until 13 dph, with higher secretion during 19–21 dph. In addition, the T3 synthesis was not inhibited by thiourea (TU) before 13 dph, but the TU response in the larvae appeared after 13 dph. Type I deiodinase (D1: GenBank accession number KC591724) was found to contain 2444 bases and encoded 248 amino acids. The D1 mRNA expression began to increase at 13 dph, with a higher expression during 15–19 dph. These results suggested that the T3 contents were maternally derived before 13 dph. Both thyroid hormonal changes and some parameters related to thyroid hormone synthesis in ontogenetic tilapia are discussed.  相似文献   

17.
A key to success in the culture of marine fish species is the mass production of high quality fry, a process largely dependent on successful first feeding and normal development and growth of fish larvae. In this regard it is important to examine the structural and functional development of the endocrine system (pituitary, thyroid, interrenal glands) during early ontogeny of marine fish. Particularly, the thyroid hormones, thyroxine (T4) and triiodothyronine (T3), influence numerous metabolic processes, such as growth, differentiation, metamorphosis, reproduction, respiration, migratory behaviour, central nervous system activity, seasonal adaptation, etc. Therefore the aim of this study was to describe the development of the thyroid gland and the ontogeny appearance of the thyroid hormones in Solea senegalensis larvae by means of histological and immunohistochemical techniques. The first thyroid follicle was present at 4 days-post-hatch (dph) coinciding with first feeding. During metamorphosis (12–20 dph) the follicles increased in both number and size, and by 30 dph presented the same characteristics as that seen in adult fish. Tissue immunostaining of both thyroid hormones decreased during the endogenous larvae development to nearly undetectable levels at the completion of yolk-sac absorption. During larvae exogenous phase, T3 and T4 immunostaining was first detected by 6 dph and an increase of specific staining for both hormones was detected between 12 and 20 dph, during metamorphosis phase.  相似文献   

18.
The digestive tract of many marine fish larvae undergoes numerous morphological and functional changes during ontogeny that can substantially influence larval survival under culture conditions. Increasing our knowledge of the digestive capacity and nutritional requirements of the larvae of new candidate species for aquaculture will aid in the development of optimal feeding protocols and greatly improve production under hatchery conditions. In this study, we assess the proteolytic capacity of California halibut (Paralichthys californicus) larvae using biochemical and histological analyses. Newly hatched larvae were reared in a semiclosed recirculating system and fed with highly unsaturated fatty acid (HUFA)–enriched rotifers from hatching until 19 d posthatch (dph) and HUFA‐enriched Artemia nauplii thereafter. Total and specific activity of trypsin and leucine‐aminopeptidase (LAP) and acid and alkaline protease activities were assessed throughout development using spectrophotometric techniques. Trypsin‐like activity and LAP and alkaline protease activities were detected shortly after hatching and before the opening of the mouth. Acid protease activity was not detected until 36–40 dph, concomitant with the development of the gastric glands. The specific activity of trypsin and LAP showed two distinct peaks at 8 and 20 dph. The second peak coincided with the shift from rotifers to Artemia. Hence, newly hatched California halibut larvae possess alkaline proteolytic activity before first feeding. Based on the digestive capacity evaluated in this study and the timing of the development of the functional stomach, we propose that California halibut can be adequately weaned to formulated microdiets around 36 dph.  相似文献   

19.
20.
黄鳝胃肠的生长抑素分泌细胞及超微结构   总被引:12,自引:1,他引:12  
舒妙安 《水产学报》2002,26(3):202-216
在肠卷石蜡法切片上,用ABC免疫组织化学方法,显示消化管全长的生长抑素分泌细胞(又称D细胞),结果只在胃体部显现,主要散在分布于胃腺中,个别出现在粘膜上皮。消化道内分泌细胞根据基部有否胞质突起,可分为4种类型,在黄鳝胃中均有,其中Ⅲ型数量最多,I、Ⅳ型次之,而Ⅱ型数量最少。由于产生生长抑素的D细胞在消化道内分泌细胞中作用多并且类型复杂,故黄鳝胃还是一个十分重要且复杂的内分泌器官。应用透射电镜,对黄鳝的胃肠作超薄切片观察显示在胃腺上皮细胞内含大量的微管泡系和一定量的酶原颗粒,表明其兼有泌酸及产生酶原的功能,属泌酸胃酶细胞。在肠上皮细胞游离面,含有大量微绒毛,但幼鳝比成鳝的长、密且整齐,提示幼鳝的肠上皮比成鳝有更强的吸收能力。  相似文献   

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