Histological study of the gastrointestinal tract in longfin yellowtail (<Emphasis Type="Italic">Seriola rivoliana</Emphasis>) larvae

This work contributes basic knowledge on larval development of Seriola rivoliana. A histological study describes the development of the digestive tract and accessory glands in S. rivoliana larvae reared under laboratory conditions at 24 °C from hatching to 30 days post-hatching (DPH). At hatching (2.6 ± 0.12 mm), larvae had an undifferentiated digestive tract with a closed straight tube and a large yolk sac with an oil globule. The liver and pancreas were observed at 1 and 2 days, and the mouth and anus opened at day 2. Enriched rotifers were visible in their digestive tract. At the beginning of the pre-flexion stage, a mixed nutritional period was observed. At day 3, exogenous feeding began; the digestive tract became differentiated into the buccopharynx, esophagus, an undifferentiated stomach, and the intestines. Zymogen granules were visible in the exocrine pancreas. At day 4, supranuclear vacuoles were present in the posterior intestine, indicating the beginning of intracellular digestion. At day 5, goblet cells were present in the esophagus and became functional at day 7 in the esophagus and intestine. The buccopharynx goblet cells developed at day 15. The presence of gastric glands and differentiation of the stomach in the fundic, cardiac, and pyloric regions during the post-flexion stage occurred at day 20. This was the onset of the juvenile period and the beginning of weaning; however, a long co-feeding phase is recommended. Pyloric caeca were observed at day 30 (13.6 ± 1.6 mm). These results provide valuable information on S. rivoliana larvae biology and digestive physiology, which should be useful to improve cultivation techniques and identify ecological features involved in ontogeny.     

Ontogeny of the digestive tract and enzymes in rock bream Oplegnathus fasciatus (Temminck et Schlegel 1844) larvae

Histological development of the digestive tract and specific activities of three digestive enzymes (trypsin, alkaline phosphatase, and pepsin) were studied in rock bream Oplegnathus fasciatus from hatching to 50 days after hatching (DAH). At hatching, the digestive tract appeared as an undifferentiated straight tube and differentiated into the buccopharynx, esophagus, stomach, intestine, and rectum at mouth opening by 3 DAH. The taste bud and mandibular teeth were present in the buccopharyx at 8 DAH. The goblet cells appeared in the esophagus at 8 DAH and in the buccopharyx at 9 DAH. The stomach anlage was formed at 2 DAH and developed into cardia, fundus, and pylorus at 14 DAH. The gastric glands were visible at 16 DAH, and the pepsin was firstly detected on 22 DAH. At 2 DAH, the intestinal valve appeared and divided the intestine into anterior intestine (AI) and posterior intestine (PI). The rectum was differentiated from the PI at 3 DAH. The supranuclear vacuoles were visible in the rectum by 6 DAH, and the lipid inclusions were present in the AI at 8 DAH. The alkaline phosphatase was detected at 1 DAH, and the increase in its activity indicated the maturation of the intestine after 40 DAH. The hepatocytes and pancreatic cells were differentiated from the blast cells at 2 DAH, and the acidophilic zymogen granules in the exocrine pancreas were observed simultaneously. The trypsin was detected by 1 DAH and increased to the maximum at 19 DAH, followed by a decrease as the stomach became functional.     

Ontogeny of the digestive tract in butter catfish Ompok bimaculatus (Bloch) larvae

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.     

Ontogenetic development of digestive functionality in golden pompano Trachinotus ovatus (Linnaeus 1758)

Ontogenetic development of the digestive system in golden pompano (Trachinotus ovatus, Linnaeus 1758) larvae was histologically and enzymatically studied from hatch to 32 day post-hatch (DPH). The development of digestive system in golden pompano can be divided into three phases: phase I starting from hatching and ending at the onset of exogenous feeding; phase II starting from first feeding (3 DPH) and finishing at the formation of gastric glands; and phase III starting from the appearance of gastric glands on 15 DPH and continuing onward. The specific activities of trypsin, amylase, and lipase increased sharply from the onset of first feeding to 5–7 DPH, followed by irregular fluctuations. Toward the end of this study, the specific activities of trypsin and amylase showed a declining trend, while the lipase activity remained at similar levels as it was at 5 DPH. The specific activity of pepsin was first detected on 15 DPH and increased with fish age. The dynamics of digestive enzymes corresponded to the structural development of the digestive system. The enzyme activities tend to be stable after the formation of the gastric glands in fish stomach on 15 DPH. The composition of digestive enzymes in larval pompano indicates that fish are able to digest protein, lipid and carbohydrate at early developmental stages. Weaning of larval pompano is recommended from 15 DPH onward. Results of the present study lead to a better understanding of the ontogeny of golden pompano during the larval stage and provide a guide to feeding and weaning of this economically important fish in hatcheries.     

The ontogeny of the alimentary tract during larval development in common pandora Pagellus erythrinus L.

The ontogenesis of the alimentary tract and its associated structures (liver, pancreas, gall bladder) was studied histologically in common pandora from hatching (0 DAH, days after hatching) until day 50 (50 DAH). Larvae were obtained by natural spawning from a broodstock adapted to captivity. They were stocked in 1500 l tanks supplied with Isochrysis galbana and Tetraselmis suecica from hatching until the Artemia feeding stage, at a temperature of 18.5–20 °C. Larvae were fed Selco-enriched Brachionus plicatilis from day 3, Artemia nauplii from day 28 and formulated feed from day 35. At hatching, the digestive tract was a histologically undifferentiated straight tube lying dorsally to the yolk sac. At first feeding (3–4 DAH) both the mouth and anus had opened and the digestive tract was differentiated into four portions: buccopharynx, oesophagus, incipient stomach and intestine. The pancreas, liver and gall bladder were also differentiated at this stage. Within 2 days after the commencement of exogenous feeding, the anterior intestinal epithelium showed large vacuoles indicating the capacity for absorption of lipids, whereas supranuclear ninhydrin-Schiff (NS) positive inclusions indicating protein absorption were observed in the posterior intestinal epithelium. Both the bile and main pancreatic ducts had opened in the anterior intestine, just after the pyloric sphincter, at this stage. Intestinal coiling was apparent since 4 DAH, while mucosal folding began at 10 DAH. Scattered PAS-positive mucous cells occurred in the oral cavity and the intestine, while they were largely diffused in the oesophagus. Gastric glands and pyloric caeca appeared at 28 DAH, indicating the transition from larval to juvenile stage and the acquisition of an adult mode of digestion.     

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.     

Nutritional regulation of intestine morphology in larval cyprinid fish, silver bream (Vimba vimba)

The present study includes the evaluation of morphological changes in the digestive tract of larval, stomachless fish silver bream ( Vimba vimba ) fed with various diets – live Artemia nauplii, commercial feed Aglo Norse (NOR) and semi-purified formulated diets: casein–gelatin (CG), dipeptide-protein (50P), dipeptide (100P), no-arginine dipeptide diet (100Pw/oArg) and a free amino acid (FAA) mixture diet. The supranuclear area of enterocytes in the posterior intestine contained enlarged absorptive vacuoles in the FAA, 100P and 100Pw/oArg groups, compared with the remaining groups. Hepatocytes' cytoplasm in fish fed with FAA, 100P and 100Pw/oArg contained mainly glycogen, and no lipid vacuoles were found. Fish fed with 100Pw/oArg showed the lowest hepatocyte surface areas while in those fed with 50P, the largest nuclei diameters were observed. Fish fed with Artemia , NOR and CG diets showed significantly ( P <0.05) higher number of proliferating cells compared with the remaining groups. Chromogranin A staining showed endocrine-immunoreactive cells (CgA-IR) in the taste buds in the oral cavity and in the enterocytes' supranuclear areas of the anterior and posterior intestine. We conclude that the growth rate and histological examination of the digestive tract in the 50P group of silver bream showed no nutritional deficiency.     

Morphogenese de l'appareil digestif et de la vessie gazeuse du turbot, Scophthalmus maximus L.Morphogenesis of the digestive system and swim bladder of the turbot, Scophthalmus maximus L.

The study is based on serial sections made on turbots raised at 18 ± 0.5°C from day 1 (post-hatching) to day 20. Histological changes of the digestive system and swim bladder were observed on a daily basis.At day 1, specimens (total length, Lt = 3.0–3.1 mm) present a digestive tract undifferentiated along its length and closed anteriorly. They belong to the embryonic period in spite of the presence of associated digestive glands (liver and pancreas).The larval period extends from day 2 to day 14–15 (Lt = 3.5–7.0 mm): the swim bladder differentiates and inflates, then the pneumatic duct binding the swim bladder to the stomach degenerates; regional differences appear along the digestive tract which starts to be functional.From day 15–16 (Lt = 6.9–7.7 mm) the juvenile period begins with the formation of gastric glands and the onset of gastric digestion. Two pyloric caeca are clearly visible and the number of intestinal folds and goblet cells increases progressively.     

Development and evaluation of microparticulate diets for early weaning of Atlantic cod Gadus morhua larvae

Early weaning trials were conducted with cod larvae to investigate the effectiveness of microparticulate diets (microbound and microcoated) with and without lipid-walled capsules (LWCs). The microparticulate diets were evaluated by measuring physical parameters of the diet in the water column (leaching and settling rate), palatability (intestinal fullness), performance of the diet (survival and growth), and examination of the diet in the larval intestine (histological analysis). A feeding trial was conducted using four experimental diets (carrageenan microbound diet, carrageenan microbound diet with LWCs, zein microcoated diet and zein microcoated diet with LWC), one commercial diet (BioKyowa: A-250) and a live feed control (rotifers and Artemia ). Survival of cod larvae to 39 days post-hatch ranged from 5 to 10% with the experimental diets, 22.9% with the BioKyowa diet, and 36.5% with live prey. There was evidence of food absorption with all diets in the form of lipid vacuoles in the midgut and supranuclear vacuoles in the hindgut. Large vacuoles in the midgut were more abundant in the enterocytes of larvae fed the experimental diets compared with larvae on the BioKyowa diet and the live feed control. Based on observations of intestinal fullness, the experimental diets appeared to be less palatable than the BioKyowa diet. As a result, it took longer to wean the larvae and higher mortality was experienced during weaning. Once successfully weaned, the experimental diets yielded growth rates equivalent to larvae feeding on the commercial diet for the remainder of the experiment.     

Ontogeny of the digestive tract of larval percula clownfish, Amphiprion percula (Lacépède 1802): a histological perspective

An understanding of the development of the digestive system of marine fish larvae is of critical importance in determining optimal feeding regimes for their culture. The present study provides information on the histomorphological development of the digestive system of clown fish, Amphiprion percula , larvae during the first month of life. Before hatching, clownfish larvae possess an alimentary tract, liver and pancreas with absorptive and digestive capabilities. The yolk sac is completely consumed within 5–7 days at 25 °C. Clownfish larvae readily accept rotifers after hatching and a complete dietary shift from rotifer to Artemia can be accomplished at 10 days after hatch (DAH). Gastric glands in the stomach first develop 11 DAH and proliferate by 15 DAH. Both non-staining vacuoles (NSV) and supranuclear inclusion vesicles (SIV) appear at 11 DAH in the midgut and hindgut respectively. Pinocytosis and extracellular digestion coexist for about 2 weeks after hatching. While SIV disappeared completely at 25 DAH, NSV continued to be a prominent feature of the midgut during the first month.     

Organogenesis of the digestive system in Pacific red snapper (Lutjanus peru) larvae

This study reports the ontogenetic development of the digestive system of larval Pacific red snapper (Lutjanus peru), an important candidate species for aquaculture on the Pacific coast of Mexico. Histological sections of larvae were cut and dyed using the haematoxylin–eosin technique. The development of the digestive tract of Pacific red snapper larvae follows a general pattern of differentiation that can be divided into three stages. Stage I lasted from 1–3 days post hatching (DPH) and included the endogenous nutrition period; it was characterized by the initial differentiation of the digestive tract in preparation for the onset of exogenous feeding (3 DPH). At this time, the digestive tract was differentiated into buccopharynx, oesophagus, stomach anlage, anterior intestine, posterior intestine and a short rectum. The liver, pancreas and kidney were also present. The mouth and anus were open. Stage II occurred after first feeding, lasted for 16 days (4–23 DPH) and included both preflexion and flexion larvae. The main changes that occurred during this stage reflected the adaptation to exogenous feeding and the concomitant growth. Stage III (24–30 DPH) included post‐flexion larvae and started with the appearance of the gastric glands and pyloric caeca. The presence of the gastric glands suggests that early weaning during culture trials of the Pacific red snapper larvae may be possible at this early age.     

Freshwater catfish jundiá (Rhamdia quelen) larvae are prepared to digest inert feed at the exogenous feeding onset: physiological and histological assessments

This study assessed the morphological development of jundiá larvae’s digestive system and digestive proteolytic activity. Specific serine proteinases activities varied over time, with the highest peak at 12 h after hatching (AH), which corresponded to 296.38 ± 84.20 mU mg^?1 for trypsin and 315.45 ± 42.16 mU mg^?1 for chymotrypsin. Specific aspartic proteinases activities increased up to the start of weaning, oscillated during that phase, but showed a consistent increase after that, resulting in the highest specific activity at 252 h AH (7.88 ± 0.68 mU mg^?1). Gel assays showed different molecular forms, especially of serine proteinases. Histology showed the gastrointestinal tract development onset at 0 h AH and open mouth at 4 h AH. At 16 h AH, the following differentiation of the digestive tract was evident: oropharyngeal cavity, esophagus, liver, pancreas, stomach, and intestine. At 40 h AH, zymogen granules in the pancreas were observed, and at 48 h AH, mucus in the digestive tract and gastric glands in the stomach. Findings indicate that jundiá has a functional stomach before the end of vitelline reserves. Therefore, jundiá larvae are probably capable to digest inert feed at the exogenous feeding onset.     

斜带石斑鱼消化系统胚后发育的组织学研究

利用形态学和连续组织切片技术，对出膜后1-60d的斜带石斑鱼各期仔鱼、稚鱼和幼鱼的消化系统进行了光镜观察，描述了其消化器官发育过程和组织学特性。研究表明，实验水温为22.0-27.8℃时，孵化后第4天，上下颌形成，卵黄囊被吸收，消化系统明显分化成食道、胃、肠、直肠以及肝脏、胆囊和胰脏等，鱼体由内源性营养转向外源性摄食营养，表明其消化系统的形态变化与食性的变化相适应。此后随着鱼体的生长，其消化系统从功能和结构上逐步完善成熟。胰脏在出膜后第4天出现，是和肝脏相互分开的一个独立的器官，但是发育到第35-60天，位于肠道后部的胰脏组织内出现许多大空泡。     

Ontogeny of the digestive tract in mud loach Misgurnus anguillicaudatus larvae

This study on histological and mucous histochemistry characteristics of the digestive system of loach (Misgurnus anguillicaudatus) was carried out from hatching (0 day after hatching, DAH) until 45 DAH. The peculiar development of both digestion processes and air‐breathing functions of the intestine was revealed. At 3 DAH, both the mouth and anus opened along with the first feed. At 4 DAH, lipid vacuoles appeared in the anterior part of the intestine and at 5 DAH the acidophilic supranuclear vacuoles appeared in the posterior part of the intestine. Mucous cells occurred in the buccopharynx and oesophagus after mouth opening and grew both in number and size as larvae grew. At 15 DAH, blood capillaries were found in the posterior part of the intestine. At 20 DAH, as a valve appeared in the intestine, the whole intestine could be divided into anterior, mid and posterior parts. With large numbers of blood capillaries and a very thin wall, a gas–blood barrier formed in the posterior intestine, which indicated that the dual roles of intestine were formed. These results suggested that the air‐breathing function of the digestive tract formed from 15 to 20 DAH, which is a critical period for loach larvae.     

Morphological and histological changes in digestive tract development during starvation in the miiuy croaker

A histological method was used to describe the ontogenetic development of the digestive tract of laboratory-reared miiuy croaker (Miichthys miiuy) and to evaluate the effects of short-term food deprivation on the morphology and histology of the digestive tract. Larvae and juveniles were maintained at 24 °C in a thermostatically controlled system. Three starvation experiments were conducted during different developmental stages: 1–7 days after hatching (dah; prior to benthic swimming); 26–35 dah (during settling); and 42–53 dah (after benthic swimming). According to the structural changes in the ontogenetic development of the digestive tract, three stages were observed. The first stage was from hatching to 3 dah; the digestive tract was undifferentiated in newly hatched larvae and then showed remarkable morphological changes and differentiation. During this period, larvae depended on endogenous nutrition. The second stage (4–20 dah) was a critical period in which larvae transitioned from endogenous feeding to exogenous feeding and the digestive tract fully differentiated into the buccopharynx, oesophagus, stomach, anterior intestine and posterior intestine. Goblet cells and vacuoles appeared in the digestive tract, and pharyngeal teeth and taste buds developed. During the third stage (20–36 dah), the gastric glands developed and the stomach differentiated into the fundic, cardiac and pyloric regions. At 25 dah, pyloric caeca developed and mucosal folds and spiral valves were clearly distinguishable. After 30 dah, the digestive tract did not undergo any noticeable differentiation, indicating the complete development of the digestive system. The wet weight and SGR (specific growth rate) of miiuy croaker larvae and juveniles greatly decreased when they were deprived of food, and compensatory growth was observed in re-feeding juveniles. The livers of starved larvae and juveniles were atrophied and dark coloured, the intestines were transparent and thin, and the stomach cubages were reduced. The histological effects of starvation were mainly evident in the degeneration of cells in digestive organs, as seen in the shrinkage and separation of cells and the loss of intercellular substances in the liver, pancreas, intestine and stomach. These changes became more severe with increased duration of starvation. In addition, the histological structure of the digestive tracts of starved larvae and juveniles partly recovered after re-feeding, and the effects of starvation on miiuy croaker were age dependent.     

Development of digestive enzyme activity in spotted rose snapper,Lutjanus guttatus (Steindachner, 1869) larvae

We describe digestive enzyme activity during the larval development of spotted rose snapper, Lutjanus guttatus. Trypsin, chymotrypsin, leucine aminopeptidase, pepsin, amylase, lipase, and acid and alkaline phosphatase activities were evaluated using spectrophotometric techniques from hatching through 30 days. The spotted rose snapper larvae present the same pattern of digestive enzyme activity previously reported for other species in which pancreatic (i.e., trypsin, chymotrypsin, amylase, and lipase) and intestinal (i.e., acid and alkaline phosphatases and leucine aminopeptidase) enzymatic activities are present from hatching allowing the larvae to digest and absorb nutrients in the yolk-sac and live prey by the time of first feeding. The digestive and absorption capacity of the spotted rose snapper increases during the larval development. A significant increase in individual activity of all enzymes occurs at 20 DAH, and around 25 DAH, the juvenile-type of digestion is observed with the appearance of pepsin secreted by the stomach, suggesting that maturation of the digestive function occurs around 20–25 DAH. Our results are in agreement with a previous suggestion that early weaning may be possible from 20 DAH. However, the patterns of enzymatic activities reported in our study should be considered during the formulation of an artificial diet for early weaning of the spotted rose snapper.     

Phospholipids in formulated start-feeds – Effect on turbot (Scophthalmus maximus L.) larval growth and mitochondrial alteration in enterocytes

Turbot Scophthalmus maximus, larvae were start‐fed with formulated feeds containing marine phospholipids (MP) or soya phospholipids (SP). The experiment was performed with six experimental groups, four groups were given formulated feeds, one group was given live feed and one group was starved. Phospholipid (PL) contents of the formulated feeds were respectively 5% MP, 15% MP, 5% SP and 15% SP. Larvae were offered feed from day 3 post‐hatch. There was no significant size difference on day 5 between the group fed 15% MP and the group given rotifers. Electron microscopical examination of enterocytes was performed on larval intestine on day 5. Larvae fed 5% and 15% MP and larvae fed rotifers had normal looking enterocytes with numerous normal looking mitochondria. In the enterocytes of larvae fed 5% SP and 15% SP the mitochondria appeared swollen with a translucent matrix and fragmented cristae. Thus, MP and not SP seemed suitable as a lipid and PL source for start‐feeding turbot larvae.     

Histology of the developing digestive system and the effect of food deprivation in larval green sturgeon (Acipenser medirostris)

The histological development of the digestive tract in hatchery-reared green sturgeon (Acipenser medirostris) larvae and the effects of food deprivation on the digestive system organization were studied from hatching until 31 days post-hatching (dph). At hatching, the larval digestive system consisted of two rudiments: a large endodermal yolk sac and a primordial hind-gut. During the endogenous feeding phase, the wall of the yolk sac differentiated into the stomach (glandular and non-glandular regions) and the anterior and intermediate intestine, while the hind-gut primordium differentiated into the spiral valve and rectum. At the onset of exogenous feeding (15 dph at 16 °C), the organization and cytoarchitecture of the digestive system in green sturgeon larvae was generally similar to those of juveniles and adults. Larvae deprived of food exhibited a progressive deterioration, with subtle pathological changes observed after 5-d starvation: shrinkage of digestive epithelia, tissue degeneration, and necrosis were observed at 10–15 d of starvation (30 dph). No changes were observed in the mucous secretion of different regions of the digestive tract of food-deprived larvae. The histological analysis of the larval digestive system may be used to evaluate the nutritional condition of larval green sturgeon in their nursery habitats in spawning rivers, which are affected by dams and flow diversions.