Changes in digestive enzyme activity during initial ontogeny of bay snook Petenia splendida

Several samples of P. splendida larvae were obtained from eggs until day 60 after hatching (dah) to determine acid and alkaline proteases, trypsin, chymotrypsin, leucine aminopeptidase, α-amylase, lipase, and acid and alkaline phosphatase activities using biochemical techniques. Additionally, SDS–PAGE alkaline protease zymogram and PAGE acid protease zymogram were carried out to identify active isoforms during larviculture. Alkaline protease and chymotrypsin were present at the moment of hatching, increased gradually reaching the maximum values at 35 dah. Trypsin and leucine aminopeptidase activities were low from hatching, increasing gradually as larvae grew. Alkaline protease zymogram showed four zymogens, which appears at different days, remaining present until the end of the larviculture (95.2 kDa at 11 dah, 26.4 kDa at 9 dah, 21.4 kDa at 3 dah, and 23.3 kDa at hatching). Pepsin activity was present at day 7 after hatching and increased progressively until the end of the larviculture. Acid protease zymogram only showed one zymogen (0.65 rf), which appear at 6 dah. Lipase was high at the time of hatching and increased until 15 dah, after which decreased gradually. Amylase was high from the beginning and until 15 dah and then decreased rapidly to almost nothing onward. Alkaline and acid phosphatases presented a high activity at the egg stage, fell slightly during the first feeding and increased again from 20 to 30 dah. Results obtained in this study show that larvae can be fed artificial diets starting on day 10 after hatching.     

Digestive enzyme activities during early ontogeny in Common snook (Centropomus undecimalis)

Common snook (Centropomus undecimalis) is one of the most important marine species under commercial exploitation in the Gulf of Mexico; for this reason, interest in developing its culture is a priority. However, larviculture remains as the main bottleneck for massive production. In this sense, our objective was to determine the changes of digestive enzymes activities using biochemical and electrophoretic techniques during 36?days of Common snook larviculture fed with live preys (microalgae, rotifers, and Artemia). During larviculture, all digestive enzymatic activities were detected with low values since yolk absorption, 2?days after hatching (dah) onwards. However, the maximum values for alkaline protease (6,500?U?mg?protein^?1), trypsin (0.053?mU?×?10^?3?mg?protein^?1), and Leucine aminopeptidase (1.4?×?10^?3?mU?mg?protein^?1) were detected at 12 dah; for chymotrypsin at 25 dah (3.8?×?10^?3?mU?mg?protein^?1), for carboxypeptidase A (280?mU?mg?protein^?1) and lipase at 36 dah (480?U?mg?protein^?1), for ??-amylase at 7 dah (1.5?U?mg?protein^?1), for acid phosphatases at 34 dah (5.5?U?mg?protein^?1), and finally for alkaline phosphatase at 25 dah (70?U?mg?protein^?1). The alkaline protease zymogram showed two active bands, the first (26.3?kDa) at 25 dah onwards, and the second (51.6?kDa) at 36 dah. The acid protease zymogram showed two bands (RF?=?0.32 and 0.51, respectively) at 34 dah. The digestive enzymatic ontogeny of C. undecimalis is very similar to other strictly marine carnivorous fish, and we suggest that weaning process should be started at 34 dah.     

Digestive enzyme activity at different developmental stages of blackspot seabream, Pagellus bogaraveo (Brunnich 1768)

Blackspot seabream, Pagellus bogaraveo (Brunnich), has been identified as a potential species to diversify European aquaculture production. Although rearing aspects have been widely investigated, little information exists on the nutritional requirements for this species. The aim of this study was to build up information on the activity of digestive enzymes at certain developmental stages of blackspot seabream in order to understand the nutritional needs of larvae and post larvae. Fish larvae were reared from hatching to 55 days after hatching (dah), and the feeding plan consisted in rotifers (5–35 dah), Artemia naupli (30–35 dah) metanaupli (35–45) and Gemma microdiet (45–55 dah). At 7, 11, 21, 45 and 55 days after hatching (dah), pooled samples of fish larvae were collected for analysis of trypsin, amylase, lipase, alkaline phosphatase and leucine–alanine peptidase activity. Up to 21 dah, the whole larvae body was used for enzymatic analysis, whereas in older larvae only the dissected abdominal cavity was used. Blackspot seabream body dry weight growth was exponential, increasing from 60 μg at 5 dah to 30±9.7 mg at 55 dah. Amylase specific activity decreased significantly during development, exhibiting at 11 dah (0.6 U mg^?1 protein) an average value 2.7 times lower than at 7 dah, and remaining stable between 45 and 55 dah (0.7 U mg protein^?1). Trypsin specific activity remained constant until 21 dah (between 38 and 44 mU mg protein^?1), which could be related to the larvae feeding regime. At later stages of development, lipase‐specific activity exhibited a significant increase (P<0.05), being three times higher at 55 dah (8 U mg protein^?1) than at 45 dah. The total activity of the studied digestive enzymes increased significantly during larval development (until 21 dah), whereas afterwards only lipase and leucine–alanine peptidase increased significantly between 45 and 55 dah. The pattern of digestive enzymes activity was related to organogenesis and the type of food used at different developmental stages.     

Development of digestive enzyme activity in larvae of spotted sand bass <Emphasis Type="Italic">Paralabrax maculatofasciatus</Emphasis>. 1. Biochemical analysis

Spotted sand bass Paralabrax maculatofasciatus is a potential aquaculture species in Northwest Mexico. In the last few years it has been possible to close its life cycle and to develop larviculture technology at on pilot scale using live food, however survival values are low (11%) and improvements in growth and survival requires the study of the morpho-physiological development during the initial ontogeny. In this research digestive activity of several enzymes were evaluated in larvae, from hatching to 30 days after hatching (dah), and in live prey (rotifers and Artemia), by use of biochemical and electrophoretic techniques. This paper, is the first of two parts, and covers only the biochemical analysis. All digestive enzyme activities were detected from mouth opening; however the, maximum activities varied among different digestive enzymes. For alkaline protease and trypsin the maximum activities were detected from 12 to 18 dah. Acid protease activity was observed from day 12 onwards. The other digestive enzymes appear between days 4 and 18 after hatching, with marked fluctuations. These activities indicate the beginning of the juvenile stage and the maturation of the digestive system, in agreement with changes that occur during morpho-physiological development and food changes from rotifers to Artemia. All enzymatic activities were detected in rotifers and Artemia, and their contribution to enhancement the digestion capacity of the larvae appears to be low, but cannot be minimised. We concluded that the enzymatic equipment of P. maculatofasciatus larvae is similar to that of other marine fish species, that it becomes complete between days 12 and 18 after hatching, and that it is totally efficient up to 25 dah.     

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.     

Histological evaluation of the elimination of Artemia nauplii from larval rearing protocols on the digestive system ontogeny of shi drum (Umbrina cirrosa L.)

The influence of the absence of Artemia nauplii from larval diet protocols on growth and digestive system ontogeny was studied using histological techniques in the shi drum (Umbrina cirrosa). One group of larvae was reared using the standard intensive rearing protocol, which offers a combination of enriched rotifers (Brachionus plicatilis), Artemia spp. nauplii and artificial diet (Std-group). Another group was reared using the same protocol, but without the offering of Artemia nauplii (group No-Artemia). The ontogenesis of the digestive system from hatching to metamorphosis was a very rapid process, and there were no differences between the two feeding regimes in the temporal appearance of the various components of the digestive system. The first organised presence of the hepatic and pancreatic tissue appeared at 2–3 d after hatching (dah), suggesting that these organs function from a very early developmental stage. In the No-Artemia larvae between 13 and 29 dah there was a reduction in the height of enterocytes in the intestinal mucosa, a progressive flattening of the primary intestinal folds in the anterior and posterior intestine and a decrease in lipid stores in the liver, suggesting a period of relative starvation. However, by the end of the study at 41 dah, there were no significant differences in body length, intestinal morphology or liver lipid stores between larvae reared under the two feeding regimes. The study suggests that the diet may influence the maturation and/or function, but not the ontogeny of the digestive system. Furthermore, the rapid differentiation of the digestive system in shi drum and the prompt recovery of the No-Artemia larvae from the symptoms of starvation by 29 dah, indicate a plasticity during ontogenesis and the ability of larvae to adapt to artificial diets at very early developmental stages.     

Ontogenic development of enzymatic activity and digestive system in Jullien's golden carp (Probarbus jullieni Sauvage, 1880)

Ontogenic development of the main enzymes and histological structure of digestive organs were studied in Jullien's golden carp (Probarbus jullieni) from hatching until 50 days after hatching (DAH). The larval fish were produced by artificial insemination and fed only Moina sp. till end of experiment. Body weight (mg) and total length (cm) of Jullien's golden carp increased exponentially and linearly. The results indicate the fish weight grew fast with increasing rate, while length increased at a constant rate over the studied period. Up‐regulation of acid protease was observed in newly hatched larvae and the specific activity gradually decreased with time. Trypsin specific activity was relatively stable within the first 35 DAH, while fluctuations in chymotrypsin were observed. Among these three proteolytic enzymes, acid proteases exihibited relatively high specific activity in newly hatched larvae, suggesting a role in yolk protein degradation. Alkaline proteases became more prominent with age and correlated with an abrupt decrease in acid proteases. Increased lipase‐specific activity appeared within 3 DAH and then gradually decreased with time, indicating the capacity to digest yolk lipid reserve. Amylase and cellulase‐specific activities changed in a similar manner, and the sensitivity to time was higher in amylase than in cellulase. The digestive organs and accessory organs developed around 3–5 DAH. However, intestinal histology was almost fully developed around 18 DAH. These findings should be useful for deciding the preferred timing for weaning, as well as on developing artificial diets referenced to the physiological changes of digestive enzymes.     

Development of digestive enzyme activity in larvae of spotted sand bass <Emphasis Type="Italic">Paralabrax maculatofasciatus</Emphasis> II: Electrophoretic analysis

The activities of several digestive enzymes during larval development of the spotted sand bass (Paralabrax maculatofasciatus) were evaluated using electrophoretic techniques. The results show the presence of three isoforms of alkaline protease from day 2 after hatching (ah) and the early appearance of one pepsin-like band from day 12 ah onwards. In addition, two lipase bands first appeared on day 2 ah, and there was a change in the molecular weight of one band from day 15 ah onwards. Several α-amylase isoforms were observed from hatching up to day 5 ah. These results indicate that the important digestive enzymes develop rapidly in these larvae, supporting the possibility of early weaning at day 12 ah using artificial diets.     

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 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 enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

Variations on development and stress defences in Solea senegalensis larvae fed on live and microencapsulated diets

Growth, development, antioxidant enzymes, stress proteins (HSP70 and HSP60), lipid peroxidation (LP) and histology in Solea senegalensis larvae were followed from 8 to 30 days post hatching (dph). Larvae were fed on three different diets: (1) live Artemia nauplii, (2) microcapsules elaborated by internal gelation, (MA) and (3) these same microcapsules but 10-fold supplemented with vitamin A (MAV). The Artemia fed group showed higher growth and a faster metamorphosis than the ones fed with microencapsulated diets, although all had similarly high survival rates of 80%. Vitamin A (VA) supplementation improved growth and development from 15 dph in relation to the strictly inert diet (MA). Larvae fed with Artemia showed organs and tissues with a normal pattern of development, whereas histological alterations were seen in larvae fed with both inert diets. The antioxidant enzymes: catalase (KAT), superoxide dismutase (SOD), and total glutathione peroxidase (t-GPX) as well as LP levels and stress proteins (HSP70 but not HSP60), measured in whole larvae, showed diet and age dependence in their response. Larvae fed with both inert diets showed similar biomarker activities, but these activities were different (p < 0.05) from larvae fed with Artemia. That is, KAT and HSP70 were lower in larvae fed with live prey and t-GPX and LP levels were lower in larvae fed with the inert food. Among the factors responsible for increased antioxidant defenses were the initiation of metamorphosis and the use of inert food. This study suggests the usefulness of the biomarkers selected as tools to evaluate the effects of compound diets on larvae.     

Egg incubation and larval rearing of navaga (Eleginus navaga Pall.), polar cod (Boreogadus saida lepechin) and arctic flounder (Liopsetta glacialis Pall.) in the laboratory

The embryonic and larval development of three White Sea cold-water fish species, rate of yolk sac absorption, age at first feeding and their survival and growth when fed different food organisms, were studied.Eggs were obtained from spawners in the Bay of Kandalaksha, White Sea, and incubated in troughs and aquaria at a mean temperature of 1.5 °C, slightly above that of the sea. The incubation period for polar cod eggs lasted 35 days, for arctic flounder, 42 days and for navaga eggs, 48 days. Emergent larvae were 5.5–6.0 mm long and began feeding at 2–4 °C, 5–6 days (navaga) and 12–14 days (polar cod) after hatching, when their yolk sac was still fairly large.They were fed day-old Artemia nauplii and zooplankton taken from the sea and consisting of Calanus and Pseudocalanus nauplii 400–600 μ in length.The period of establishing first feeding is the most critical for larvae.     

Digestive enzyme activity during early larval development of the Cuban gar Atractosteus tristoechus

The ontogenesis of digestive enzymes (proteases, amylases, lipases, and phosphatases) in Cuban gar Atractosteus tristoechus was determined in larvae between 5 and 18 days after hatching (DAH). Variations in specific activities of most enzymes were related to the transition from endogenous to exogenous feeding and to the transition from the larval to the juvenile stage. Alkaline protease activity was not detected until 8 DAH in contrast to acid protease activity, which was quantifiable at 5 DAH. Acid protease activity was consistently higher than alkaline protease activity, indicating the presence of a functional stomach in the early stages of larval development. The acid protease activities of larvae and adults were compared by means of zymogram analysis. Four acid protease bands were found in adults (two more than in larvae). This result is the first time that more than one band of acid proteolytic activity has been found in Lepisosteidae. High lipase activity indicated the importance of lipid utilization, particularly during yolk-sac absorption. In contrast to the other enzymes studied, amylase activity was consistently low, probably due to the strictly carnivorous diet of gar larvae and their low capacity for carbohydrate digestion. High activities of aminopeptidase and acid and alkaline phosphatases suggest intestinal absorption. This result, together with the existence of a short gut and a lower proteolytic activity in the intestine than in the stomach, suggest that most of the proteolytic activity takes place in the stomach, while the primary function of the intestine is nutrient uptake.     

黄条鰤仔稚幼鱼消化酶活性变化研究

为了解黄条鰤(Seriola aureovittata)早期发育阶段的消化生理特性,测定了黄条鰤胚胎、仔稚幼鱼阶段脂肪酶、淀粉酶、胰蛋白酶和碱性磷酸酶活性变化。结果显示,在黄条鰤仔鱼出膜前胚胎阶段,即能检测到脂肪酶、淀粉酶和碱性磷酸酶活性;初孵仔鱼体内(1 d)初次检测出胰蛋白酶的活性。脂肪酶和碱性磷酸酶比活力在仔鱼孵化后迅速增强(P<0.05),在4 d开口时,2种酶比活力达最高值;淀粉酶比活力在7 d时达最大值;胰蛋白酶比活力在仔鱼阶段缓慢上升,15 d时比活力最大。稚鱼阶段内脏团中脂肪酶、碱性磷酸酶和胰蛋白酶活性基本维持稳定,幼鱼阶段内脏团脂肪酶、碱性磷酸酶和胰蛋白酶活性都呈现上升趋势;稚鱼和幼鱼阶段内脏团中淀粉酶活性下降并基本稳定于较低水平。研究表明,黄条鰤仔稚幼鱼发育过程中,各种消化酶活性变化明显,且与其发育阶段和食性密切相关。在尚未摄食饵料的早期仔鱼体内已存在消化酶,认为其是母源传递而来,不是由外源性饵料所致;幼鱼阶段内脏团脂肪酶、碱性磷酸酶和胰蛋白酶比活力明显提高,这反映出随苗种生长发育,其肠道结构和消化机能逐渐完善,并且对脂肪、蛋白质的需求逐渐增强。     

Substrate-SDS-PAGE determination of protease activity through larval development in sea bream

Identification of alkaline proteases produced during larval stages of gilthead sea bream, Sparus aurata was achieved using SDS-PAGE and specific inhibitors. Such techniques were also applied to determine proteases existing in rotifers, Brachionus plicatilis, and Artemia nauplii, which are used as live food for these larvae, as well as proteases of adult fish. The results show a great prominence of trypsin-like proteases during the 4 weeks after hatching, but the number of enzyme species was reduced in adult fish. Alkaline proteases present in the rotifers and Artemia showed clear differences when compared with those of the larvae and were not detected in extracts obtained from fed larvae. The results obtained provide information about the role of exogenous enzymes in larval feeding of sea bream.     

黄条仔稚幼鱼消化酶活性变化研究

为了解黄条(Seriola aureovittata)早期发育阶段的消化生理特性,测定了黄条胚胎、仔稚幼鱼阶段脂肪酶、淀粉酶、胰蛋白酶和碱性磷酸酶活性变化。结果显示,在黄条仔鱼出膜前胚胎阶段,即能检测到脂肪酶、淀粉酶和碱性磷酸酶活性;初孵仔鱼体内(1 d)初次检测出胰蛋白酶的活性。脂肪酶和碱性磷酸酶比活力在仔鱼孵化后迅速增强(P<0.05),在4 d开口时,2种酶比活力达最高值;淀粉酶比活力在7 d时达最大值;胰蛋白酶比活力在仔鱼阶段缓慢上升,15 d时比活力最大。稚鱼阶段内脏团中脂肪酶、碱性磷酸酶和胰蛋白酶活性基本维持稳定,幼鱼阶段内脏团脂肪酶、碱性磷酸酶和胰蛋白酶活性都呈现上升趋势;稚鱼和幼鱼阶段内脏团中淀粉酶活性下降并基本稳定于较低水平。研究表明,黄条仔稚幼鱼发育过程中,各种消化酶活性变化明显,且与其发育阶段和食性密切相关。在尚未摄食饵料的早期仔鱼体内已存在消化酶,认为其是母源传递而来,不是由外源性饵料所致;幼鱼阶段内脏团脂肪酶、碱性磷酸酶和胰蛋白酶比活力明显提高,这反映出随苗种生长发育,其肠道结构和消化机能逐渐完善,并且对脂肪、蛋白质的需求逐渐增强。     

Diurnal variation of tryptic activity in larval stage and development of proteolytic enzyme activities of malabar grouper (Epinephelus malabaricus) after hatching

Diurnal variation in tryptic activity and developmental changes in proteolytic enzyme activities of malabar grouper larvae (Epinephelus malabaricus) were examined. Five different groups were prepared for the experiment of diurnal variation of tryptic activity in larvae: larvae were fed Thai-type rotifers Brachionus rotundiformis from the time of mouth opening, fed rotifers from 6 h after mouth opening, 12 h, 24 h and not fed rotifers (starved control). The experimental tanks were placed in temperature-controlled baths at 28 °C under 24 h light. Developmental changes in proteolytic activity of trypsin and pepsin-like enzyme were measured from hatching to 57 days after hatching (DAH).

The tryptic activity of all fed groups showed the same pattern, and the diurnal variation of tryptic activity was clearly observed from 3 to 6 DAH. The highest tryptic activities were found at 19:00, and the activities were lowest from 01:00 to 07:00. In contrast, that of non-fed larvae was low compared to the fed groups, however the diurnal variation of tryptic activity was shown same tendency to the fed groups. Interestingly, both groups (fed and non-fed) were exhibited a circadian rhythm under the 24 h light conditions and delaying of first-feeding. Tryptic activity of larvae notably increased from 40 to 45 DAH and markedly decreased at 52 DAH. In contrast to the tryptic activity, that of pepsin-like enzyme clearly increased from 47 to 51 DAH. The results suggest that a functional change of protein digestion occurs from 40 to 50 DAH related with metamorphosis in malabar grouper. These results could contribute to determining appropriate feeding schedules, such as feeding time, frequency and optimal time to change food items, in mass-scale production of the present species.     

Larval rearing of African carp,Labeo parvus Boulenger, 1902 (Pisces: Cyprinidae), using live food and artificial diet under controlled conditions

This study aimed at (1) evaluating the efficacy of live food organisms (Artemia and natural zooplankton) and an artificial diet in the larval rearing of African carp Labeo parvus and (2) determining appropriate rearing conditions. After yolk sac resorption, the larvae were separated into five different feeding trials with two replicates. At the end of the larval rearing period (28 days post‐hatching), the highest (94.6%) and the lowest (53.7%) cumulative survival rates were found when larvae were fed with natural zooplankton for 7 days followed by Nippai food for 21 days, and when larvae were fed from the beginning of exogenous feeding with Nippai food only respectively. The significant highest body weight (351.6 mg), total length (34.4 mm) and specific growth rates (15.5%day^?1) were recorded when the larvae were fed with Artemia nauplii for 14 days followed by Nippai food for 14 days. The lowest growth performance (body weight and specific growth rates) were obtained when larvae were fed exclusively Nippai food. These results indicate that L. parvus can be successfully cultured in indoor nursery systems from hatching to the early juvenile stage.