Visualization of musculature of the larval greater amberjack Seriola dumerili (Risso) using whole‐mount immunostaining with special reference to cranial muscles

Muscles play important roles in feeding, respiration and swimming during not only adult stage but also larval stage in fish. However, there is no information of the muscle development in larval greater amberjack, Seriola dumerili (Risso). Here, we investigated muscle development in the greater amberjack, focusing primarily on the cranial muscles from 0 days post hatch (dph) to 12 dph using a modified whole‐mount immunohistochemical staining method. We found that the muscles required for feeding develop by 3 dph, when the larvae begin to feed. Subsequently, muscle composition in the dorsal branchial arches changes to the adult form between 5 and 8 dph. At 8 dph, all the muscles required for feeding and respiration appear, whereas the dorsal, pelvic and caudal fin muscles required for swimming develop later. This report provides fundamental information on larval greater amberjack muscle development, which will enable the detection of abnormal larval muscles and improve larval rearing techniques by modifying environmental conditions.     

Effect of temperature on survival,growth and malformation of cultured larvae and juveniles of the seven-band grouper Epinephelus septemfasciatus

The effects of rearing temperature (23–29 °C) during the larval and juvenile stage on survival, growth and skeletal malformations in the seven-band grouper Epinephelus septemfasciatus were investigated. The survival rate of juveniles 30–40 mm in total length emerging from eggs was higher at 25 and 26 °C (0.1–1.3 %) than at 23 °C or 27–29 °C (0.004–1.5 %). Growth (increase in total length) was accelerated at higher temperatures. The frequency of malformed individuals was lower at 25–27 °C (36.0–61.5 %) than at 23, 28 or 29 °C (65.3–76.9 %). Specific incidences of spinal curvature and centrum fusion or defects in juveniles were not related to rearing temperature. However, incidences of twisted or compressed vertebrae (6.5–64.0 %) were higher at higher temperatures, while the incidence of bifurcated neural spine was significantly higher at 23 °C (43.6–54.4 %) than at other temperatures (3.3–22.7 %). The incidence rate of spinal curvature (23.3 %) was significantly higher in juveniles with a deflated swim bladder, regardless of rearing temperature. The results of this study suggest that the optimum culture temperature for seven-band grouper is 25–26 °C, collectively considering the survival, growth and incidences of abnormalities. Our results also demonstrate the significance of identifying the conditions for swim bladder inflation to prevent spinal curvature in seven-band grouper.     

Early-weaning diets for gilthead sea bream (Sparus aurata L.) and their potential use in Hellenic marine fish hatcheries

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.     

Effect of the early temperature on the growth of larvae and postlarvae turbot, <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> L.: muscle structural and ultrastructural study

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.     

Rearing temperature affects Senegalese sole (Solea senegalensis) larvae protein metabolic capacity

The present work examined the short- and long-term effects of three rearing temperatures on protein metabolism and growth trajectories of Senegalese sole larvae using ¹⁴C-labelled Artemia protein as feed. A first feeding trial was performed on larvae reared at 15, 18 and 21 °C (at 26, 17 and 14 days post-hatching (dph), respectively) and a second trial conducted on post-larvae after transfer to the same rearing temperature (~20 °C) (49, 35 and 27 dph, in larvae initially reared at 15, 18 and 21 °C, respectively). Temperature greatly influenced larvae relative growth rate (RGR) and survival, since growth at 15 °C was severely depressed. Protein digestibility and retention was highest at 18 °C during the first trial (85.35 ± 1.16 and 86.34 ± 2.33 %, respectively). However, during the second trial, post-larvae from 15 °C had the highest feed intake and protein digestibility (3.58 ± 1.54 and 75.50 ± 1.35 %, respectively), although retention was similar between treatments. Furthermore, after transfer to 20 °C larvae from 15 °C experienced compensatory growth, which was observed until 121 dph, and confirmed by RGR values, which were significantly higher at 15 ºC than at 21 ºC or 18 ºC. Results from the present study show that Solea senegalensis larval development, survival and protein digestion and retention are highly affected by thermal history.     

Effect of diet and rearing conditions on growth and survival throughout developmental stages of larvae and juveniles of dusky grouper <Emphasis Type="Italic">Epinephelus marginatus</Emphasis> (Lowe 1834)

Two rearing conditions, tank light-dimensions and diet, were tested in dusky grouper—larvae and juveniles—in order to improve larval rearing techniques for this species. Larvae reared in high-volume and fluorescent tubes showed the highest survival rate at 18 dph (17.67%, 16%). This might be due to a higher depth and less trapping area during surface death in high-volume tanks than in low volume. Besides, a 500-lx evenly distributed light with fluorescent tubes might improve survival rate at early stages rather than illumination with energy-efficient light bulbs (4.68%, 9.64%). Light bulbs created shaded areas with less illumination inducing low larval activity. First feeding was performed with minimum rotifer lorica width of 102?±?10.2 μm. No mixed diet was supplied. Throughout metamorphosis, stress shock syndrome appeared to be the main cause of mortality. Diet A showed best growth and survival (4.1–7.36%) during this period probably due to its high HUFA content (9.5%). Contrary to what was observed during first stages, groupers showed best performance with lower illumination. Cannibalism was observed during this period in all culture conditions.     

The influence of feeding diets containing wheat gluten supplemented with dipeptides or free amino acids on structure and development of the skeletal muscle of carp (Cyprinus carpio)

A 28-day feeding trial was conducted to evaluate the influence of feeding diets containing plant protein wheat gluten supplemented with dipeptides or free amino acids on structure and development of the skeletal muscles of carp (Cyprinus carpio). Common carp fingerlings (1 month old) having an of average weight of 0.07 ± 0.02 g and total length of 17.79 ± 1.79 mm were fed three formulated diets—wheat gluten protein-based diets supplemented with Lys–Gly dipeptide (PP), free lysine and glycine (AA), control diet without lysine supplemented (CON)—and two other diets: restrictive diet—frozen zooplankton (Z) and commercial diet Aglo Norse (AN). After 28 days of experimental feeding, statistically significant higher survival was observed among fish fed AN and Z diets (99.5 ± 1.0 %; P ≤ 0.05). The feeding AN diet has had also a positive influence on weight and growth rate as well as on development and growth of skeletal muscles. Furthermore, carps fed AN diet had the largest area of red and white muscle as compared with the other feeding groups, and the differences were statistically significant (P ≤ 0.05). The increase in the number of proliferating cells (proliferating cell nuclear antigen) was observed on the last day of the experiment among carps fed PP, AA and CON. Moreover, fish fed PP significantly had the greatest number of MyoD- and myogenin-positive nucleus (P ≤ 0.05). Among the experimental diets based on wheat gluten, a positive impact on structure and development of muscles has been observed in carps fed PP diet.     

Early development of the shortfin silverside Chirostoma humboldtianum (Valenciennes, 1835) (Atheriniformes: Atherinopsidae)

The shortfin silverside Chirostoma humboldtianum has been considered for culture in Mexico, but success has been limited by a poor knowledge of its early development. First synthesis of the early development of the shortfin silverside is presented to determine conditions suitable for rearing. Brooder maturation was induced through photothermal cycles. C. humboldtianum ova were fertilized in vitro. The eggs were incubated in reconstituted water (160-180 mg/L CaCO₃) at 18 °C and 5 gm of NaCl per litre. During the hatching day, 300 shortfin silversides were stocked and followed up until metamorphosis in order to establish the timing of exogenous feeding, changes in food type, growth and development during critical periods for survival, according to the theory of saltatory ontogeny. Free embryos hatched 12 days after fertilization at 18 °C. First critical point for survival is the beginning of exogenous feeding. Free embryos started mixed feeding on day four of post-hatching (dph), point of no-return was presented towards the end of mixed feeding on 6 dph, larval period began at six (dph) when the anus is opened, and metamorphosis to juvenile was presented at 65 dph with a SL of 19.34 ± 2.28 mm, when scales and fins were well developed. Differences in growth between periods were detected: free embryos growth slower than larvae but mouth size depicted a larger growth rate in the former. Cephalic length and mouth size were negatively related to standard length in embryos and larvae. Mouth size was positively related to cephalic length in free embryos but negative in larvae. Results suggest that during the free embryo phase, growth priorities are directed to the development of apparatuses and systems; whereas, during the larval period, energy is directed to growth in length, mouth size and development of fins, which allows them to increase their swimming velocity, grants them a greater capacity to obtain exogenous food and, in consequence, increases fitness for survival.     

美洲黑石斑鱼(Centropristis striata)消化系统胚后发育的组织学观察

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.     

Muscle growth in full-cycle cultured Pacific bluefin tuna Thunnus orientalis from early larval to juvenile stage: histochemical,immunohistochemical and ultrastructural observation

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.     

Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae

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.     

Larval development in European hake (Merluccius merluccius L.) reared in a semi-intensive culture system

Eggs of European hake (Merluccius merluccius L.) were stripped from fish caught at sea. Larvae were kept under semi‐intensive conditions at around 12°C. In addition, eggs were incubated in single wells at 9.2, 12.7 and 14.5°C, where hatching, development and survival were closely examined. During the larval stage, a total of 299 larvae were sampled to follow development and growth. In addition a small number of juveniles were sampled. Larvae hatched approximately 4 days after fertilization, and were 2.9 mm in total length (TL). At 6‐day post hatching (dph), the larvae were 4.1 mm (TL), the jaw apparatus was developed, and the larvae had started to feed. Most of the growth during the early larval period is restricted to the head, and there is almost no increase in length for the first 3–4 weeks post hatching. Teeth and pelvic fins appear at 25 dph. Development of unpaired fins at approximately 30 dph marks the start of the larval–juvenile transition. Weaning to formulated feed was accomplished 50 dph, when external morphology was similar to that of adult hake.     

Morphological development and allometric growth of yellowtail kingfish Seriola lalandi V. larvae under culture conditions

Morphological development and allometric growth patterns of Seriola lalandi larvae were assessed to characterize normal growth patterns under culture conditions. Early ontogenetic stages of yellowtail kingfish exhibited an exponential growth in terms of standard length as a function of age. Five development stages were characterized from hatching to the juvenile stage: larval stage I (0–2 days post hatch, dph) with endogenous feeding, characterized by a small yolk sac, unpigmented eyes, primordial finfold surrounding the body and a closed mouth; larval stage II (2–15 dph) characterized by mouth opening, complete pigmentation of eyes and the beginning of the exogenous feeding; subsequently, in the larval stage III (15–25 dph) the posterior tip of notochord of the larvae bended upward and the first rays appeared in fins, concomitant with a change in swimming behaviour; thereafter, larval stage IV (post‐flexion stage; 25–30 dph) began when larvae resembled in morphology to a juvenile organism; however, caudal and dorsal fins were not completely development. Lastly, the juvenile stage was reached 30 dph characterized by a morphology and fin structures similar to those of the adults. Growth and development of structures and organs associated with vital functions such as feeding, sensorial and breathing systems seemed to be more critical previous to 23 dph, which was reflected with a positive allometric growth of head and eyes during this period. The results from this study can be used as a tool‐guide to assess normal development in larval research with S. lalandi to improve existing rearing protocols in hatchery production.     

Activities of digestive enzymes and histology of digestive system during larval development of devil stinger (Inimicus japonicus)

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).     

The Effect of Temperature on First Feeding, Growth, and Survival of Larval Witch Flounder Glyptocephalus cynoglossus

Witch flounder Glyptocephalus cynoglossus has recently been identified as a candidate species for aquaculture in the northeastern United States and the Canadian Atlantic Provinces. This study investigated the optimal temperatures for witch flounder larval first feeding and for long term larval culture from hatching through metamorphosis. Maximum first feeding occurred between 15.0 and 16.2 C. Larvae did not survive beyond first feeding when reared at mean temperatures of 5.1, 10.4, or 19.5 C and were unable to initiate feeding at mean rearing temperatures below 6.0 C. At a rearing temperature of 15.0 C in 16-L tanks, mean larval survival to 60 days post hatch (dph) was 14.1%. Mean overall length-specific growth rate for larvae reared to 60 dph at 15.0 C was 3.5%/d and mean absolute growth was 0.62 mm/d. Subsequent larval growth at 15.6 C began to taper off towards 70 dph at the onset of weaning which overlapped with larval metamorphosis. Growth plateaued at 85 dph, followed by a rebound between 90 and 95 dph. Survival was 100% when weaning onto a dry, pelleted diet was initiated at 70 dph with a 10-d live diet co-feeding period. These results are favorable and encourage the further pursuit of commercial witch flounder culture.     

The effect of different rotifer feeding regimes on the growth and survival of yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae

First feeding success is critical to larval marine finfish and optimization of live feed densities is important for larval performance and the economics of commercial hatchery production. This study investigated various rotifer feeding regimes on the prey consumption, growth and survival of yellowtail kingfish Seriola lalandi larvae over the first 12 days post hatch (dph). The common practice of maintaining high densities of rotifers (10–30 ind. mL^?1) in the rearing tank was compared to a low density feeding technique, where 5–8 ind. mL^?1 of rotifers were offered. A ‘hybrid’ feeding regime offered rotifers at the high density treatment until 5 dph and the lower feeding densities thereafter. There was no significant difference in larval survival (hybrid: 28.9 ± 7%, low density: 17.3 ± 5% and high density: 17.2 ± 9%) or growth (hybrid: 6.12 ± 0.18 mm, low density: 6.03 ± 0.10 mm and high density: 6.11 ± 0.23 mm) between treatments. Rotifer ingestion was independent of rotifer density throughout the trial and increased with larval age, with larvae at 4 dph ingesting 22 ± 1.5 rotifers larvae^?1 h^?1 and by 11 dph ingesting 59 ± 1.6 rotifers larvae^?1 h^?1. These data demonstrate that from first feeding, yellowtail kingfish larvae are efficient at capturing prey at the densities presented here and consequently significant savings in rotifer production costs as well as other potential benefits such as facilitation of early weaning and improved rotifer nutritional value may be obtained by utilizing lower density rotifer feeding regimes.     

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.     

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.     

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 .