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.     

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.     

Mass Production of Juveniles of the Fat Snook Centropomus parallelus in Brazil

A pilot‐scale trial to rear fat snook Centropomus parallelus through larval, weaning and nursery phases was conducted in Florianópolis, Brazil. Eggs (96% fertilization) from captive broodstock, induced to spawn using 50 μg/kg LHRHa were stocked in two 4,000‐L cylindrical fiberglass tanks at a mean density of 19.2 eggs/L. Nannochloropsis oculata was stocked with the eggs and maintained at a mean density of 0.5–1.0 ± 10⁶ cells/mL up to 31 dph (31 dph). Hatching averaged 90%. Larvae were fed rotifers Brachionus rotundiformis enriched with an oil emulsion from 3 dph to 36 dph (30–40 rotifers/mL) and Artermia meta‐nauplii enriched with Selco from 22 dph to 60 dph (mean 2.9 meta‐nauplii/mL). Weaning began at 45 dph with an artificial dry diet NRG (50% protein), supplied together with concentrated and enriched Artemia meta‐nauplii. No critical period of mortality was observed during larval rearing. During the 43 days of weaning and nursery, less than 1% mortality was recorded. Food conversion rate during nursery was 1.17, with a change in the coefficient of variation of mean total length of 1.3%. Specific total growth rate in weight was 13.0 %/d and mean growth in total length and total weight were 0.65 mm/d and 24.0 mg/d over the whole rearing trial respectively. Mean total length and total weight of juveniles were 57.6 ± 0.1 mm and 2.11 ± 0.12 g, respectively, and the length‐weight relationship was W = 8.29931 ± 10–⁵ TL^3.049607 (r= 0.9986). on 88 dph when the trial was terminated. The condition factor on 88 dph was 1.104. On 88 dph a total of 35,000 juveniles were harvested, overall survival was 25.5% with mean final density of 4.4 fishn and biomass of 6.9 kg/m³. The present trial demonstrated the feasibility of mass production of fat snook. Possible improvements necessary for commercial cultivation of fat snook C. parallelus are discussed.     

Ontogeny of the digestive tract of Arapaima gigas (Schinz, 1822) (Osteoglossiformes: Arapaimidae) larvae

Early‐life survival of Arapaima gigas is one of the main challenges of its farming. In this study, we described the morphological and histochemical development of the gastrointestinal tract of arapaima larvae. Larvae were collected from a pond when they started to swim to the water surface (initial day) and were housed in indoor tanks. Daily samplings (n = 10) were performed from 0 to the 11th day after the collection (DAC) and then on the 14th, 17th, and 20th DAC. On the initial day, arapaima larvae (0.05 ± 0.01 g; 2.21 ± 0.06 cm) had opened mouth and anus and no yolk sac. In addition, larvae presented well‐developed digestive organs. Gastric glands were fully formed, with positive reactions to alcian blue (AB) pH 1.0 as well as to periodic acid‐Schiff (PAS) in the simple columnar epithelium. There were folds throughout the intestine and brush border, with an AB pH of 2.5 + PAS positive mucins. From 1 to 11 DAC, larvae presented increasing concentrations of gastric glands and thickness of the stomach muscular layer. From 14 to 20 DAC, the intestine presented high‐fold complexity. We suggest that arapaima larvae may be fed exogenous inert diets at a size of around 2 cm.     

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.     

Ontogenetic development of digestive tract and digestive enzymatic activities in Squaliobarbus curriculus larvae

Squaliobarbus curriculus is an economically important freshwater fish. The ontogenetic development of the digestive system of S. curriculus larvae was studied histologically and enzymatically from hatching to 30 days posthatching (DPH). Amylase, lipase, alkaline phosphatase and pepsin activities were detected from the hatching stage, indicating that these enzymes were genetically preprogrammed. Marked increases in intestinal amylase, trypsin and alkaline phosphatase activities between 10 and 20 DPH corresponded to feed acquisition and transformation. Larval development in S. curriculus could be divided into three phases: phase I (endotrophic period): 1–3 DPH; phase II (endo‐exotrophic period): 4–5 DPH; and phase III (exclusively exotrophic period): from 6 DPH onward. At hatching, the digestive tract of the larvae was an undifferentiated straight tube. On 3 DPH, the digestive tract differentiated into the mouth cavity, oesophagus and intestine. On 6 DPH, feeding was totally exotrophic and the yolk sac was completely exhausted. During the growth of S. curriculus larvae, the intestinal mucosa formed and the number of goblet cells and microvilli increased, demonstrating maturation of the digestive system. The study about the digestive development of S. curriculus larvae will contribute to better larval‐rearing strategies.     

Compensatory growth responses in juvenile fat snook,Centropomus parallelus Poey,following food deprivation

The compensatory growth in juvenile fat snook (13.7 g) was evaluated in fish subjected to four treatments: continuously fed (control); subjected to 1 (D1), 2 (D2) and 3 (D3) weeks of food deprivation, followed by a 5‐week re‐feeding period. At the end of food deprivation, the body weights at the different treatments were significantly different and inversely related to the length of the food deprivation period (C=16.425 g; D1=14.024 g; D2=13.542 g; D3=12.228 g). During re‐feeding, no differences were observed in the slopes of body weight growth curves among treatments, indicating failure in full growth compensation. In the first re‐feeding week (week 4), although hyperphagia was observed for all re‐fed groups, better food conversion ratios were detected only for D1 and D2, reflected in an enhancement in the specific growth rates. From week 5 onwards, no differences between treatments and control were detected for any variable. The total food intake was lower for D2 and D3 compared with the control. In the present study, fish showed partial compensatory growth when subjected to a maximum of 2 weeks of food deprivation. Therefore, the food deprivation can promote production losses, even considering a reduction in the total food intake.     

中国龙虾早期叶状幼体消化道的组织结构观察

利用光镜和电镜对中国龙虾叶状幼体发育早期(Ⅰ～Ⅳ期)消化道的组织学、组织化学和超微结构进行了研究。消化道可分为前肠、中肠和后肠。前肠包括口、食道和胃。消化道壁由粘膜层、结缔组织层、肌肉层和外膜组成。除中肠外,其余消化管壁上皮均覆盖有几丁质层。消化道结构随幼体发育逐渐复杂化。口部的口器发达。食道壁内褶形成四个食道嵴。贲门胃的结构简单,至Ⅳ期幼体只有胃磨齿的雏形;幽门胃内具有由栉状刚毛组成的腺滤器,其结构与过滤功能逐渐完善。中肠上皮细胞具微绒毛,胞质中富含胞器。后肠有六个纵嵴。各段管壁上皮细胞内的胞器以线粒体与内质网居多。在各期幼体消化道中,几丁质层具较多的多糖类物质,中肠上皮细胞含少量糖原;肌细胞内含丰富蛋白质,幽门胃与中肠上皮的含量次之。还探讨了龙虾幼体消化道的结构与功能的关系。     

Histological and histochemical changes in the digestive tract of white sturgeon larvae during ontogeny

Ontogenetic changes in digestive tract histology and digestive enzyme histochemistry were investigated 11 to 36 days post-hatch in white sturgeon Acipenser transmontanus larvae. From initiation of exogenous feeding (12 days post-hatch), larvae were fed a commercial salmonid diet for the ensuing 24 days. The digestive system of white sturgeon displayed a high degree of morphologic organization and functionality at the onset of exogenous feeding. An enhancement of digestive capacities occurred with transition to active feeding. On day 2 of feeding, there was a clear increase of alkaline phosphatase, aminopeptidase M, dipeptidyl peptidase IV, and -glutamyl transpeptidase activity in the brush border of the spiral intestine. This strong activity is an apparent confirmation of the importance of this segment of the intestine for protein digestion and nutrient absorption. The functional development of the pyloric intestine occurred on day 4 and was concomitant with an increase in the activity of brush border and cytoplasmic enzymes such as acetylcholinesterase, dipeptidyl peptidase II, - and -galactosidases. The absence of acetylcholinesterase, lactase, nonspecific esterase, and weak activity of exopeptidases and alkaline phosphatase in the anterior intestine suggests that this segment of the intestine may be less important in nutrient absorption than the pyloric and spiral intestines. The observed quantitative and qualitative differences in enzyme activity along the intestine indicate a high degree of specialization of each segment for specific digestive and absorptive processes.     

Ontogeny of the digestive tract of thick lipped grey mullet (Chelon labrosus) larvae reared in “mesocosms”

This work describes the ontogeny of the digestive tract in thick lipped grey mullet (Chelon labrosus) larvae reared until day 36 post-hatching with the semi-extensive technology in mesocosms. Diet was constituted by live preys, rotifers, Artemia and wild zooplankton, then compound diet was added from day 20 (p. h.). Linear growth, weight growth and digestive enzymes specific activities were studied during larval ontogeny. Pancreatic enzymes (trypsin and amylase) and intestinal enzymes (leucine-alanine peptidase “Leu-ala”, aminopeptidase N “AN” and alkaline phosphatase “AP”) were assayed in larvae sampled throughout the rearing trial to evaluate gastrointestinal maturation along the development.The trypsin specific activities were very high during the first two weeks and then declined as observed in marine fish species. A following increase in trypsin specific activity from day 20 was attributed notably to ingestion of particle compound diet. In contrast to the pattern generally described in fish larvae, amylase specific activity showed a continuous increase. This could be attributed to the fact that C. labrosus is an omnivorous species and suggests that the fish might be able to use efficiently diets containing higher levels of starch or other carbohydrates since the end of larval development.Relative expression of intestinal brush border membrane enzymes (AP and AN) and cytosolic enzyme (Leu-ala), showed an abrupt increase of both AP/leu-ala and AN/leu-ala ratios at day 8 (p. h.), indicating that maturation of intestinal tract in C. labrosus larvae is particularly precocious. It is assumed that larvae of C. labrosus might support early co-feeding and weaning strategies, which could reasonably be initiated since mouth opening.     

Ontogeny and distribution of alkaline and acid phosphatases in the digestive system of California halibut larvae (Paralichthys californicus)

Studies aimed to assess the digestive physiology of marine fish larvae under culture conditions are important to further understand the functional characteristics and digestive capacities of the developing larvae. Most studies to date concentrate on intestinal lumen digestion and little attention to the absorption process. Thus, the objectives of this study were to histochemically detect and quantify some of the enzymes responsible for absorption and intracellular digestion of nutrients in the anterior and posterior intestine of California halibut larvae. Alkaline and acid phosphatases were detected from the first days post-hatch (dph). Alkaline phosphatase maintained a high level of activity during the first 20 dph in both intestinal regions. Thereafter, a clear intestinal regionalization of the activity was observed with the highest levels occurring in the anterior intestine. Acid phosphatase activity gradually increased in both intestinal regions during development, and a regionalization of the activity was not observed until late in development, once the ocular migration began. Highest levels were observed in the anterior intestine at the end of metamorphosis concomitant with the stomach development. The results from this study show some morphological and physiological changes are occurring during larval development and a clear regionalization of the absorption process as the larvae develops. These ontological changes must be considered in the elaboration of diets according to the digestive capacity of the larvae.     

Effects of stocking density and size distribution on growth, survival and cannibalism in juvenile fat snook (Centropomus parallelus Poey)

This study was carried out in order to determine the effect of stocking density and size distribution on growth, survival and cannibalism of fat snook (Centropomus parallelus Poey) juveniles. Three stocking densities (1.5, 3 and 6 fish L⁻¹) were tested in two groups of fish: homogeneous [total length (TL)=22.1 mm; coefficient of variation of length (CV)=8%] and heterogeneous (TL=22.7 mm; CV=15%). The experimental design was factorial (3 × 2) using triplicates. Fish were reared for 30 days in 80‐L circular tanks with an open flow‐through system. There was no significant interaction between the two factors (density and size distribution). The highest cannibalism rate (14.2%) was observed in the heterogeneous group, which also had a significantly higher growth rate (TL=45.6 mm) than the homogeneous group (3.2% and TL=40.9 mm). Significant differences were found between the two lowest densities and the highest density (6.2%, 6.3% and 13.4% respectively). Growth was not affected by fish density. Cannibalism was the main cause of mortality. In 90% of the occurrences, whole prey ingestion was observed. According to the results obtained, cannibalism was positively correlated to density, although size grading of fat snook juveniles is recommended to control cannibalism in all the densities tested.     

Post-prandial protease activity in the digestive tract of African catfish Clarias gariepinus larvae fed decapsulated cysts of Artemia

The alkaline proteolytic activity in the gut of African catfish larvae was studied during short time ranges from 30 min to 4 h after ingestion of decapsulated Artemia cysts. The variation in total protease and trypsin activities during the day was monitored during starvation, after one single meal ingestion, and during continuous feeding. In starved larvae the enzymatic activity was low and did not change in time. No significant endogenous secretion of digestive enzymes was detected. The level of alkaline proteolytic activity found in starved larvae was further considered as the basal level. In larvae fed only one meal during the day, the enzyme activity significantly increased from 3 h post-feeding up to a maximum level found 12 h after feeding. In the larvae receiving a meal every 4 h, the effect of feeding on the proteolytic activity was significantly different from the one in fish fed only once a day. The total protease activity in this dietary treatment changed according to the time of feeding and fluctuated around a constant level, which was intermediate between the maximum and the basal level. No rhythmic cycle of enzyme production in the fish was observed when the proteolytic activity was studied during a cycle of 24 h. When specific trypsin activity was measured, a similar pattern was found as with the total protease. The contribution of digestive enzymes from Artemia to the total digestion of food by the catfish larvae was calculated to be less than 1% of the total amount of the proteolytic activity measured in the larval gut.     

Ontogeny of the digestive tract in shi drum (Umbrina cirrosa L.) reared using the mesocosm larval rearing system

Histological changes of the digestive tract were studied in shi drum (Umbrina cirrosa) from hatching until 41 days post hatching (dph), when the fry had a mean (±S.D.) total length (TL) of 32 ± 2 mm and wet weight (WW) of 0.42 ± 0.07 g. Larvae were reared using the mesocosm technique, the most natural among commercially employed rearing methods for marine larvae. Shi drum opened their mouth at 2 dph (2.78 ± 0.09 mm TL), at which time 90% of the larvae already had an inflated swim bladder. The differentiation of the digestive tract into buccopharynx, esophagus, and anterior and posterior intestine was completed by 3 dph (2.82 ± 0.07 mm TL), 1 day after the onset of exogenous feeding. The alimentary canal started coiling and formed its first loop at 2 dph, while the pancreas and liver were differentiated at 3 dph. Yolk sac reserves lasted until 7 dph (4.3 ± 0.1 mm TL), suggesting a brief period of endogenous and exogenous feeding. The first esophageal goblet cells appeared at 7 dph containing acid mucins and at 8 dph taste buds appeared on the buccopharyngeal epithelium. The stomach was morphologically differentiated at 9 dph (5.5 ± 0.1 mm TL) when gastric glands became abundant in the cardiac region, and the first pyloric caeca appeared at 14 dph (10.1 ± 0.9 mm TL). Supranuclear eosinophilic vacuoles were observed in the posterior intestine between 3 and 11 dph (6.3 ± 0.9 mm TL). Their number decreased as the stomach differentiated, suggesting a change in the protein digestion mechanism. The results of the study suggest a rapid development of shi drum and its digestive system and underline the possibility of weaning larvae to artificial feed even earlier than the 12 dph employed in the present study.     

Characterization of the digestive tract of wild ayu

ABSTRACT: The gut and gut contents of ayu Plecoglossus altivelis caught in the Ohta River in Hiroshima prefecture, Japan were examined. Relative intestine length (intestine length/body length) was almost constant. Except for relative intestine length, characteristics of the gut of wild ayu were not significantly different from those of cultured ayu. The height of intestinal folds increased and their width declined from stomach to anus. Cyanobacteria ( Homoeathrix sp. and Calothrix sp.) and diatoms ( Cymbella sp., Gomphonema sp., Melosira sp., Navicula sp., and Synedra sp.) were well digested during passage through the gut despite a lack of apparent destruction. In contrast, green algae ( Dictyosphearium sp., Coelastrum sp., and Pediastrum sp.) exhibited little degradation. The pH of stomach contents ranged from 2.8 to 7.4 (mean ± SD = 4.1 ± 1.0), whereas that of the posterior intestine was 7.0–8.5 (7.9 ± 0.4). Gut contents of wild ayu increased with bodyweight, and were more than threefold greater than those of cultured ayu, suggesting that wild ayu compensate for low nutrient content of algae through the continuous ingestion of large quantities of feed organisms.     

甲壳动物幼体消化酶研究进展

通过研究近年来国内外学者关于甲壳动物幼体消化酶方面的研究现状,发现目前已测定出甲壳动物幼体消化酶的种类主要有：蛋白酶、胰蛋白酶、羧肽酶A、羧肽酶B、氨肽酶、胶原酶、弹性蛋白酶、胃蛋白酶、糜蛋白酶、酯酶、脂肪酶、淀粉酶、纤维素酶、几丁质酶、麦芽糖酶、蔗糖酶、昆布多糖酶等种类,初步归纳了甲壳动物幼体主要消化酶的分子量、最适pH和最适温度;分析了幼体发育过程中消化酶活力变化模式以及与食性的关系,并采用淀粉酶/蛋白酶活力（A/P）比值或淀粉酶/类胰蛋白酶（A/T）比值作为甲壳动物幼体的食性指标;观察了幼体消化器官的发育,阐明幼体消化酶的合成与分泌不仅由遗传控制、中肠腺发育程度等因子决定,而且还与饲料的营养水平有关,提出采用幼体消化酶对饲料组成的适应性研究幼体营养需求的新途径.[中国水产科学,2006,13（3）：492-501]     

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.